CONJUGATES OF CHEMOTHERAPY AGENTS AND TISSUE-BINDING SMALL MOLECULES, COMPOSITIONS AND METHODS THEREOF

Abstract

The invention provides novel conjugates of tissue-binding small molecules and therapeutic agents and pharmaceutical compositions thereof, and their use in and methods of treatment of certain diseases or conditions (e.g., cancer).

Claims

1. A compound having the structural formula of (I) or (II): ##STR00108## wherein X is CHR or NR, and R is H or alkyl, ##STR00109## wherein R.sup.1 is methyl, or a pharmaceutically acceptable form or an isotope derivative thereof.

2. The compound of claim 1, selected from: ##STR00110##

3. (canceled)

4. The compound of claim 1, selected from: ##STR00111##

5. (canceled)

6. A compound having the structural formula (III) or (IV): ##STR00112## wherein n is an integer selected from 2-8, ##STR00113## wherein n is an integer selected from 2-10, or a pharmaceutically acceptable form or an isotope derivative thereof.

7. The compound of claim 6, selected from: ##STR00114##

8. (canceled)

9. The compound of claim 7, selected from: ##STR00115## ##STR00116##

10. A compound having the structural formula (V): ##STR00117## wherein R.sup.2 is H or P(O)R.sub.2, and R is alkyl, L is a single bond or a group selected from: ##STR00118## and k is an integer selected from 0-4, ##STR00119## wherein R.sup.2 is H or P(O)R.sub.2, and R is alkyl, L is a single bond or a group selected from: ##STR00120## and k is an integer selected from 0-4, ##STR00121## ##STR00122## or a pharmaceutically acceptable form or an isotope derivative thereof.

11. The compound of claim 10, selected from: ##STR00123##

12. (canceled)

13. The compound of claim 10, having the structure: ##STR00124##

14-19. (canceled)

20. A pharmaceutical composition comprising a compound according to claim 1 and a pharmaceutically acceptable excipient, carrier, or diluent.

21. The pharmaceutical composition of claim 20, effective to treat or reduce cancer, or a related disease or condition.

22. A unit dosage form comprising a pharmaceutical composition according to claim 20.

23. A method for treating or reducing a disease or condition, comprising administering to a subject in need thereof a pharmaceutical composition comprising a compound according to claim 1 and a pharmaceutically acceptable excipient, carrier, or diluent.

24. The method of claim 23, wherein the disease or condition is cancer, or a related disease or condition thereof.

25-27. (canceled)

Description

DETAILED DESCRIPTION OF THE INVENTION

[0044] The invention provides novel small molecule compounds, methods of their synthesis, and pharmaceutical compositions as well as methods thereof for treating or reducing various diseases or conditions.

[0045] A central feature of the present invention is that compounds of the invention are cancer-targeting and slow-releasing therapeutic agents, affording targeted and sustained delivery. The conjugates of the invention are comprised of a tissue protein binder, a cleavable linker and a small molecule drug. After local injection of the conjugate, the tissue protein binder, acting as a molecular glue, binds to tissue proteins in solid tumor, thereby retaining the conjugates in the solid tumor without leaking to systemic circulation. The small molecule drug is then slowly released from the conjugate by breakage of the cleavable linker. Slow but sustained release of the drug inside the solid tumor amplifies the tumor-killing effect while minimizing the adverse reaction because minimum amount of the drug is leaked into systemic circulation. The dosing schedule can be varied depending on the half-life of the conjugates.

[0046] In one aspect, the invention generally relates to a compound having the structural formula of (I):

##STR00010##

wherein [0047] X is CHR or NR, and [0048] R is H or alkyl,
or a pharmaceutically acceptable form or an isotope derivative thereof.

[0049] In certain embodiments, the compound of formula (I) is selected from:

##STR00011##

[0050] In another aspect, the invention generally relates to a compound having the structural formula of (II):

##STR00012##

or a pharmaceutically acceptable form or an isotope derivative thereof.

[0051] In certain embodiments, the compound of formula (II) is selected from:

##STR00013##

[0052] In yet another aspect, the invention generally relates to a compound having the structural formula of:

##STR00014##

or a pharmaceutically acceptable form or an isotope derivative thereof.

[0053] In yet another aspect, the invention generally relates to a compound having the structural formula of (III):

##STR00015## [0054] wherein n is an intergern selected from 2-8 (i.e., 2, 3, 4, 5, 6, 7 or 8), [0055] or a pharmaceutically acceptable form or an isotope derivative thereof.

[0056] In certain embodiments, the compound of formula (III) is selected from:

##STR00016##

[0057] In yet another aspect, the invention generally relates to a compound having the structural formula of (IV).

##STR00017## [0058] wherein n is an intergern selected from 2-10 (e.g., 2, 3, 4, 5, 6, 7, 8, 9 or 10), [0059] or a pharmaceutically acceptable form or an isotope derivative thereof.

[0060] In certain embodiments, the compound of formula (IV) is selected from:

##STR00018## ##STR00019##

[0061] In yet another aspect, the invention generally relates to a compound having the structural formula of (V):

##STR00020##

wherein [0062] R.sup.2 is H or (O)(R.sup.3).sub.2 and R.sup.3 is alkyl, [0063] L is a single bond or a group selected from:

##STR00021## [0064] k is an integer selected from 0-4 (e.g., 0, 1, 2, 3 or 4), or a pharmaceutically acceptable form or an isotope derivative thereof.

[0065] In certain embodiments, the compound of formula (V) is selected from:

##STR00022##

[0066] In yet another aspect, the invention generally relates to a compound having the structural formula of (VI):

##STR00023##

wherein [0067] R.sup.2 is H or (O)(R.sup.3).sub.2 and R.sup.3 is alkyl, [0068] L is a single bond or a group selected from:

##STR00024##

and [0069] k is an integer selected from 0-4 (e.g., 0, 1, 2, 3 or 4), or a pharmaceutically acceptable form or an isotope derivative thereof.

[0070] In certain embodiments, the compound of formula (VI) has the structure:

##STR00025##

[0071] In yet another aspect, the invention generally relates to a compound having the structural formula:

##STR00026##

or a pharmaceutically acceptable form or an isotope derivative thereof.

[0072] In yet another aspect, the invention generally relates to a compound having the structural formula:

##STR00027##

or a pharmaceutically acceptable form or an isotope derivative thereof.

[0073] In yet another aspect, the invention generally relates to a compound having the structural formula:

##STR00028##

or a pharmaceutically acceptable form or an isotope derivative thereof.

[0074] In yet another aspect, the invention generally relates to a compound having the structural formula:

##STR00029##

or a pharmaceutically acceptable form or an isotope derivative thereof.

[0075] In yet another aspect, the invention generally relates to a compound having the structural formula:

##STR00030##

or a pharmaceutically acceptable form or an isotope derivative thereof.

[0076] In yet another aspect, the invention generally relates to a compound having the structural formula:

##STR00031##

or a pharmaceutically acceptable form or an isotope derivative thereof.

[0077] In yet another aspect, the invention generally relates to a pharmaceutical composition comprising a compound disclosed herein and a pharmaceutically acceptable excipient, carrier, or diluent.

[0078] In certain embodiments, the pharmaceutical composition of the invention is effective to treat or reduce cancer, or a related disease or condition.

[0079] In yet another aspect, the invention generally relates to a unit dosage form comprising a pharmaceutical composition comprising a compound disclosed herein.

[0080] In yet another aspect, the invention generally relates to a method for treating or reducing a disease or condition, comprising administering to a subject in need thereof a pharmaceutical composition comprising a compound disclosed herein and a pharmaceutically acceptable excipient, carrier, or diluent.

[0081] In certain embodiments, the disease or condition is cancer, or a related disease or condition thereof.

[0082] In yet another aspect, the invention generally relates to use of a compound disclosed herein for treating or reducing a disease or condition, for example, cancer.

[0083] In yet another aspect, the invention generally relates to use of a compound disclosed herein, and a pharmaceutically acceptable excipient, carrier, or diluent, in preparation of a medicament for treating or reducing a disease or condition, for example, cancer.

[0084] Compositions of the present invention are administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir. The term parenteral as used herein includes subcutaneous, intravenous, intramuscular, intra-articular, intra-synovial, intrasternal, intrathecal, intrahepatic, intralesional and intracranial injection or infusion techniques. Preferably, the compositions are administered orally, intraperitoneally or intravenously. Sterile injectable forms of the compositions of this invention include aqueous or oleaginous suspension. These suspensions are formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation is also a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that are employed are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium.

[0085] For this purpose, any bland fixed oil employed includes synthetic mono- or di-glycerides. Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically-acceptable oils, such as olive oil or castor oil, especially in their polyoxyethylated versions. These oil solutions or suspensions also contain a long-chain alcohol diluent or dispersant, such as carboxymethyl cellulose or similar dispersing agents that are commonly used in the formulation of pharmaceutically acceptable dosage forms including emulsions and suspensions. Other commonly used surfactants, such as Tweens, Spans and other emulsifying agents or bioavailability enhancers which are commonly used in the manufacture of pharmaceutically acceptable solid, liquid, or other dosage forms are also be used for the purposes of formulation.

[0086] Pharmaceutically acceptable compositions of this invention are orally administered in any orally acceptable dosage form. Exemplary oral dosage forms are capsules, tablets, aqueous suspensions or solutions. In the case of tablets for oral use, carriers commonly used include lactose and corn starch. Lubricating agents, such as magnesium stearate, are also typically added. For oral administration in a capsule form, useful diluents include lactose and dried cornstarch. When aqueous suspensions are required for oral use, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening, flavoring or coloring agents are optionally also added.

[0087] Alternatively, pharmaceutically acceptable compositions of this invention are administered in the form of suppositories for rectal administration. These can be prepared by mixing the agent with a suitable non-irritating excipient that is solid at room temperature but liquid at rectal temperature and therefore will melt in the rectum to release the drug. Such materials include cocoa butter, beeswax and polyethylene glycols.

[0088] Pharmaceutically acceptable compositions of this invention are also administered topically, especially when the target of treatment includes areas or organs readily accessible by topical application, including diseases of the eye, the skin, or the lower intestinal tract. Suitable topical formulations are readily prepared for each of these areas or organs.

[0089] Topical application for the lower intestinal tract can be effected in a rectal suppository formulation (see above) or in a suitable enema formulation. Topically-transdermal patches are also used.

[0090] For topical applications, provided pharmaceutically acceptable compositions are formulated in a suitable ointment containing the active component suspended or dissolved in one or more carriers. Exemplary carriers for topical administration of compounds of this are mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax and water. Alternatively, provided pharmaceutically acceptable compositions can be formulated in a suitable lotion or cream containing the active components suspended or dissolved in one or more pharmaceutically acceptable carriers. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.

[0091] Pharmaceutically acceptable compositions of this invention are optionally administered by nasal aerosol or inhalation. Such compositions are prepared according to techniques well-known in the art of pharmaceutical formulation and are prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and/or other conventional solubilizing or dispersing agents.

[0092] Most preferably, pharmaceutically acceptable compositions of this invention are formulated for oral administration. Such formulations may be administered with or without food. In some embodiments, pharmaceutically acceptable compositions of this invention are administered without food. In other embodiments, pharmaceutically acceptable compositions of this invention are administered with food.

[0093] The amount of compounds of the present invention that are optionally combined with the carrier materials to produce a composition in a single dosage form will vary depending upon the host treated, the particular mode of administration. Preferably, provided compositions should be formulated so that a dosage of between 0.01-100 mg/kg body weight/day of the compound can be administered to a patient receiving these compositions.

[0094] It should also be understood that a specific dosage and treatment regimen for any particular patient will depend upon a variety of factors, including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, and the judgment of the treating physician and the severity of the particular disease being treated. The amount of a compound of the present invention in the composition will also depend upon the particular compound in the composition.

EXAMPLES

[0095] As depicted in the Examples below, in certain exemplary embodiments, compounds are prepared according to the following general procedures. It will be appreciated that, although the general methods depict the synthesis of certain compounds of the present invention, the following general methods, and other methods known to one of ordinary skill in the art, can be applied to all compounds and subclasses and species of each of these compounds, as described herein.

[0096] Compound numbers utilized in the Examples below correspond to compound numbers set forth supra.

[0097] .sup.1H was recorded at 400 MHz on a Varian Mercury 400 spectrometer. .sup.13C NMR was recorded at 100 MHz. Proton chemical shifts were internally referenced to the residual proton resonance in CDCl3 (7.26 ppm). Carbon chemical shifts were internally referenced to the deuterated solvent signals in CDCl3 (77.20 ppm).

[0098] LC-MS spectra were recorded on a Shimadzu LC-MS2020 using Agilent C18 column (Eclipse XDB-C18, 5 m, 2.150 mm) with flow rate of 1 mL/min. Mobile phase A: 0.1% of formic acid in water; mobile phase B: 0.1% of formic acid in acetonitrile. A general gradient method was used.

TABLE-US-00001 Time (min) A B 0 95 5 3 0 100 4 0 100 4.05 95 5

[0099] Analytical HPLC was performed on Agilent 1200 HPLC with a Zorbax Eclipse XDB C18 column (2.1150 mm) with flow rate of 1 mL/min. Mobile phase A: 0.1% of TFA in water; mobile phase B: 0.1% of TFA in acetonitrile. A general method with the following gradient was used.

TABLE-US-00002 Time (min) Mobile Phase A Mobile Phase B 0 95 5 15 0 100 16 0 100 16.5 95 5 16.5 stop

[0100] Preparative HPLC was performed on Varian ProStar using Hamilton C18 PRP-1 column (15250 mm) with flow rate of 20 mL/min. Mobile phase A: 0.1% of TFA in water; mobile phase B: 0.1% of TFA in acetonitrile. A typical gradient method was used.

TABLE-US-00003 Time (min) Mobile Phase A Mobile Phase B 0 90 10 30 30 70 35 10 100 40 90 10 45 stop

Example 1

(R)-4,11-diethyl-4-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano [3,4:6,7]indolizino[1,2-b]quinolin-9-yl 4-(4-iodophenyl)butanoate (1)

##STR00032##

##STR00033##

[0101] Step 1: To a solution of compound 1-2 (600 mg, 2.06 mmol) in DMF (10 mL) was added EDCI (800 mg, 4.18 mmol) and HOBt (190 mg, 1.40 mmol). After stirring for 5 minutes, compound 1-1 (SN-38, 500 mg, 1.27 mmol) was added, and the mixture was stirred at room temperature for 16 hours. After completion of the reaction, water (100 mL) was added and the mixture was stirred at room temperature for 30 min. A white precipitation was formed completely. The resulting mixture was filtered and washed with water to give an off-white solid. The crude product was then triturated by acetonitrile for one hour and filtered. The filter cake was washed with acetonitrile and dried to give the title compound 1 (505 mg, 59% yield) as an off-white solid. .sup.1H NMR (400 MHz, DMSO-d6) 8.19 (d, J=9.1 Hz, 1H), 8.04-7.94 (m, 1H), 7.66 (td, J=10.1, 9.2, 4.6 Hz, 3H), 7.32 (s, 1H), 7.10 (d, J=7.8 Hz, 2H), 6.53 (s, 1H), 5.44 (s, 2H), 5.32 (s, 2H), 3.17 (q, J=7.6 Hz, 2H), 2.76-2.62 (m, 4H), 1.98 (p, J=7.4 Hz, 2H), 1.87 (hept, J=7.0 Hz, 2H), 1.28 (t, J=7.5 Hz, 3H), 0.88 (t, J=7.3 Hz, 3H). LCMS: m/z calculated for C.sub.32H.sub.29IN.sub.2O.sub.6: 664.50; found: 665.2 [M+H].sup.+.

Example 2

(R)-4,11-diethyl-4-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano [3,4:6,7]indolizino[1,2-b]quinolin-9-yl 2-methyl-4-(4-iodophenyl)butanoate (2)

##STR00034##

##STR00035##

[0102] Step 1: LDA (2 M, 2.5 mL, 5 mmol) was added dropwise to a stirring solution of compound 1-2 (500 mg, 1.72 mmol) in anhydrous THF (10 mL) at 0 C. under nitrogen atmosphere. After addition, the mixture was stirred at 0 C. for 30 min. Mel (538 mg, 3.79 mmol) was added dropwise and the reaction was allowed to stir at room temperature for 16 h. After completion of the reaction, the mixture was quenched with saturated NH.sub.4Cl (30 mL) solution and extracted with EtOAc (30 mL3). The organic layers were washed with brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The crude product was purified by silica gel column (eluted with 30% EtOAc in Petroleum ether) to give the title compound 2-1 (400 mg, yield 76%).

[0103] Step 2: A mixture of compound 2-1 (390 mg, 1.24 mmol), compound 1-1 (194 mg, 0.496 mmol), HOBt (335 mg, 2.48 mmol), DMAP (30 mg, 0.248 mmol) and EDCI (950 mg, 4.97 mmol) in DMF (5 mL) and DCM (5 mL) was stirred at room temperature for 16 h. After completion of the reaction, the mixture was partitioned between EtOAc (100 mL) and water (100 mL). The organic layer was washed with brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The crude product was purified by silica gel column (eluted with 5% MeOH in DCM) to afford the title compound 2 (265 mg, yield 30%).

Example 3

(S)-4,11-diethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano [3,4:6,7]indolizino[1,2-b]quinolin-9-yl(4-iodophenethyl)carbamate (3)

##STR00036##

##STR00037##

[0104] Step 1: To a solution of compound 3-1 (1.0 g, 4.11 mmol) in anhydrous THE (10 mL) at 0 C. was added dropwise a solution of borane-tetrahydrofuran complex (1 M in THF, 18.5 mL) under nitrogen atmosphere. The reaction mixture was stirred at 65 C. for 2 h. After completion of the reaction, the mixture was cooled to 0 C. and quenched by addition of 6 N HCl (2 mL). The resulting mixture was then basified with 1 N NaOH solution and extracted with DCM (50 mL2). The combined organic layers were dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to afford the title compound 3-2 (1.0 g, yield 90%) as a colorless oil, which was used directly at the next step without further purification.

[0105] Step 2: Compound 3-3 (345 mg, 1.71 mmol) was dissolved in DCM (5 mL) and cooled to 0 C. To this was added slowly a solution of compound 3-2 (210 mg, 0.85 mmol) and TEA (257 mg, 2.55 mmol) in DMSO (2 mL). The mixture was stirred at 0 C. for 1 h. SN-38 (1-1, 330 mg, 0.84 mmol) and TEA (100 mg, 0.99 mmol) were added. Then the mixture was stirred at room temperature for 3 h. After completion of the reaction, the mixture was partitioned between EtOAc (50 mL) and water (50 mL). The organic layer was washed with brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The crude product was purified by silica gel column (eluted with 5% MeOH in DCM) to afford the title compound 3 (120 mg, yield 20%).

Example 4

(S)-4,11-diethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3,4:6,7]indolizino[1,2-b]quinolin-9-yl(4-iodophenethyl)(methyl)carbamate (4)

##STR00038##

##STR00039##

[0106] Step 1: A mixture of compound 3-2 (1.0 g, 4.04 mmol), BOC.sub.2O (1.7 g, 7.79 mmol) and TEA (830 mg, 8.20 mmol) in DCM (10 mL) was stirred at room temperature for 2 h. After completion of the reaction, the mixture was concentrated. The residue was purified by silica gel column (elute with 10% EtOAc in petroleum ether) to afford the title compound 4-1 (1.1 g, yield 80%) as a white solid. LCMS: m/z calculated for C.sub.13H.sub.18INO.sub.2: 347.20; found: 348.21 [M+H].sup.+.

[0107] Step 2: NaH (60% in mineral oil, 450 mg, 11.25 mmol) was added in several portions to a solution of compound 4-1 (1.3 g, 3.74 mmol) in anhydrous DMF (5 mL) at 0 C. After stirring for 1 h, Mel (1.6 g, 11.27 mmol) was added to the mixture in one portion. Then the mixture was stirred at room temperature for 16 h. After completion of the reaction, the mixture was quenched by addition of ice water and partitioned between EtOAc (100 mL) and water (50 mL). The organic layer was washed with brine, dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The crude product was purified by silica gel column (elute with 5% EtOAc in petroleum ether) to afford the title compound 4-2 (1.02 g, yield 75%) as a white solid. LCMS: m/z calculated for C.sub.14H.sub.20INO.sub.2: 361.22; found: 306.04 [M-.sup.tBu+H].sup.+.

[0108] Step 3: A mixture of compound 4-2 (1.02 g, 2.82 mmol) in EtOAc (5 mL) and a solution of HCl in EtOAc (4 N, 5 mL) was stirred at room temperature for 1 h. After completion of the reaction, a white precipitation was formed completely. The white precipitation was filtered and washed with EtOAc to afford the title compound 4-3 (610 mg, yield 82%), which was used directly at the next step without further purification.

[0109] Step 4: A suspension of SN-38 (1-1, 250 mg, 0.637 mmol), DIPEA (170 mg, 1.31 mmol) and compound 4-4 (215 mg, 0.706 mmol) in anhydrous DMF (5 mL) was stirred at 0 C. under nitrogen atmosphere for 1 h. Then the mixture was treated with a solution of compound 4-3 (227 mg, 0.763 mmol) and DIPEA (200 mg, 1.55 mmol). After stirring for an additional 2 h. The resulting mixture was diluted with EtOAc (50 mL) and washed with water. The organic layer was washed with brine, dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The crude product was purified by silica gel column (elute with 2% MeOH in DCM) to afford 4 (170 mg, yield 39%) as a light-yellow solid. .sup.1H NMR (400 MHz, Chloroform-d) 8.37-8.16 (m, 1H), 7.84 (d, J=2.8 Hz, 1H), 7.69 (dd, J=5.6, 3.0 Hz, 3H), 7.61-7.48 (m, 1H), 7.35-7.28 (m, 1H), 7.16-6.97 (m, 2H), 5.79 (dd, J=16.3, 2.8 Hz, 1H), 5.49-5.26 (m, 3H), 3.91 (s, 1H), 3.80 (t, J=7.1 Hz, 1H), 3.66 (t, J=7.8 Hz, 1H), 3.53 (d, J=2.8 Hz, 1H), 3.19 (d, J=7.4 Hz, 1H), 3.16 (d, J=2.8 Hz, 1H), 3.09 (d, J=2.8 Hz, 1H), 3.01-2.91 (m, 2H), 1.93 (ddd, J=15.0, 7.4, 4.0 Hz, 2H), 1.48-1.38 (m, 3H), 1.13-1.02 (m, 3H). LCMS: m/z calculated for C.sub.32H.sub.30IN.sub.3O.sub.6: 679.51; found: 680.31 [M+H].sup.+.

Example 5

(S)-4,11-diethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano [3,4:6,7]indolizino[1,2-b]quinolin-9-yl 2-(4-isobutylphenyl)propanoate (5)

##STR00040##

##STR00041##

[0110] Step 1: A mixture of compound 5-1 (236.6 mg, 1.15 mmol), EDCI (438.1 mg, 2.29 mmol) and HOBt (103.2 mg, 0.76 mmol) in DMF (3 mL) was stirred at room temperature for 5 min. Compound 1-1 (300 mg, 0.76 mmol) was added and the mixture was stirred at room temperature for 15 h. The mixture was partitioned between EtOAc (30 mL) and water (30 mL). The organic layer was washed with brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The crude product was purified by silica gel column (eluted with 2% MeOH in DCM) to afford the title compound 5 (324 mg, yield 72%) as a white solid. .sup.1H NMR (500 MHz, DMSO-d.sub.6) 8.18 (d, J=9.1 Hz, 1H), 7.89 (d, J=2.4 Hz, 1H), 7.50 (dd, J=9.2, 2.5 Hz, 1H), 7.39-7.36 (m, 2H), 7.32 (s, 1H), 7.22-7.19 (m, 2H), 6.53 (s, 1H), 5.43 (s, 2H), 5.31 (s, 2H), 4.15 (q, J=7.1 Hz, 1H), 3.15 (q, J=7.6 Hz, 2H), 2.46 (d, J=7.2 Hz, 2H), 1.92-1.81 (m, 3H), 1.57 (d, J=7.1 Hz, 3H), 1.27 (t, J=7.6 Hz, 3H), 0.88 (dd, J=7.1, 2.3 Hz, 9H). LCMS: m/z calculated for C.sub.35H.sub.36N.sub.2O.sub.6: 580.68; found: 581.47 [M+H].sup.+.

Example 6

(S)-4,11-diethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano [3,4:6,7]indolizino[1,2-b]quinolin-9-yl(S)-2-(6-methoxynaphthalen-2-yl)propanoate (6)

##STR00042##

##STR00043##

[0111] Step 1: A mixture of compound 6-1 (176.0 mg, 0.76 mmol), SN-38 (1-1) (150 mg, 0.38 mmol), HOBt (51.6 mg, 0.38 mmol) and EDCI (219.0 mg, 1.15 mmol) in DMF (3 mL) was stirred at room temperature for 9 h. After completion of the reaction, the reaction mixture was diluted with EtOAc and washed with water. The organic layer was washed with brine, dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The crude product was purified by column (eluted with 2% MeOH in dichloromethane) to afford 6 (185 mg, yield 80%) as a white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 8.16 (d, J=9.1 Hz, 1H), 7.91 (dd, J=4.4, 2.1 Hz, 2H), 7.87 (d, J=8.7 Hz, 2H), 7.58 (dd, J=8.5, 1.8 Hz, 1H), 7.52 (dd, J=9.1, 2.5 Hz, 1H), 7.34 (d, J=2.6 Hz, 1H), 7.30 (s, 1H), 7.19 (dd, J=9.0, 2.5 Hz, 1H), 6.53 (s, 1H), 5.43 (s, 2H), 5.30 (s, 2H), 4.32 (q, J=7.1 Hz, 1H), 3.87 (s, 3H), 3.14 (q, J=7.5 Hz, 2H), 1.85 (dq, J=14.2, 7.1 Hz, 2H), 1.66 (d, J=7.1 Hz, 3H), 1.25 (t, J=7.6 Hz, 3H), 0.87 (t, J=7.3 Hz, 3H). LCMS: m/z calculated for C.sub.36H.sub.32N.sub.2O.sub.7: 604.66; found: 605.36. [M+H].sup.+. TLC (DCM:MeOH/40:1): R.sub.f (compound 6-1)=0.5; R.sub.f (compound 1-1)=0.3; R.sub.f (6)=0.37.

Example 7

(S)-4,11-diethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano [3,4:6,7]indolizino[1,2-b]quinolin-9-yl(R)-2-(4-isobutylphenyl)propanoate (7)

##STR00044##

##STR00045##

[0112] Step 1: A mixture of compound 7-1 (107 mg, 0.51 mmol), EDCI (198.2 mg, 1.04 mmol) and HOBt (46.7 mg, 0.35 mmol) in DMF (3 mL) was stirred at room temperature for 5 min. Compound 1-1 (135.7 mg, 0.35 mmol) was added and the mixture was stirred at room temperature and for 15 h. The reaction mixture was partitioned between EtOAc (20 mL) and water (20 mL). The organic layer was washed with brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The crude product was purified by silica gel column (eluted with 2% MeOH in DCM) to afford the title compound 7 (161 mg, yield 80%) as a white solid. .sup.1H NMR (500 MHz, DMSO-d.sub.6) 8.22-8.11 (m, 1H), 7.93-7.86 (m, 1H), 7.53-7.46 (m, 1H), 7.38 (d, J=7.6 Hz, 2H), 7.32 (d, J=2.3 Hz, 1H), 7.21 (d, J=7.8 Hz, 2H), 6.53 (s, 1H), 5.44 (s, 2H), 5.33 (s, 2H), 4.16 (q, J=7.2 Hz, 1H), 3.16 (p, J=6.6, 6.1 Hz, 2H), 2.47 (d, J=7.2 Hz, 2H), 1.93-1.80 (m, 3H), 1.57 (d, J=7.1 Hz, 3H), 1.27 (t, J=7.6 Hz, 3H), 0.88 (d, J=6.3 Hz, 9H). LCMS: m/z calculated for C.sub.35H.sub.36N.sub.2O.sub.6: 580.68; found: 581.47 [M+H].sup.+.

Example 8

(S)-4,11-diethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano [3,4:6,7]indolizino[1,2-b]quinolin-9-yl(S)-2-(4-isobutylphenyl)propanoate (8)

##STR00046##

##STR00047##

[0113] Step 1: A mixture of compound 8-1 (157.7 mg, 0.76 mmol), EDCI (292.0 mg, 1.53 mmol) and HOBt (68.8 mg, 0.51 mmol) in DMF (2 mL) was stirred at room temperature for 5 min. Compound 1-1 (200 mg, 0.51 mmol) was added and the mixture was stirred at room temperature for 15 h. The reaction mixture was partitioned between EtOAc (30 mL) and water (30 mL). The organic layer was washed with brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The crude product was purified by silica gel column (eluted with 2% MeOH in DCM) to afford the title compound 8 (197 mg, yield 66%) as a white solid. .sup.1H NMR (500 MHz, DMSO-d.sub.6) 8.17 (d, J=9.1 Hz, 1H), 7.89 (t, J=2.3 Hz, 1H), 7.50 (dd, J=9.1, 2.5 Hz, 1H), 7.39-7.36 (m, 2H), 7.31 (s, 1H), 7.22-7.19 (m, 2H), 6.52 (s, 1H), 5.43 (s, 2H), 5.31 (s, 2H), 4.15 (q, J=7.1 Hz, 1H), 3.15 (q, J=7.6 Hz, 2H), 2.46 (d, J=7.1 Hz, 2H), 1.92-1.80 (m, 3H), 1.57 (d, J=7.1 Hz, 3H), 1.26 (t, J=7.6 Hz, 3H), 0.87 (dd, J=7.0, 2.8 Hz, 9H). LCMS: m/z calculated for C.sub.35H.sub.36N.sub.2O.sub.6: 580.68; found: 581.47 [M+H].sup.+.

Example 9

N-((S)-1-(((R)-1-(6-(2-(4-(4-iodophenyl)butanamido)ethyl)-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)-3-methylbutyl)amino)-1-oxo-3-phenylpropan-2-yl)pyrazine-2-carbox amide (9)

##STR00048##

##STR00049##

[0114] Step 1: To a solution of compound 9-1 (1.0 g, 6.24 mmol) in DMF (25 mL) was added ethyl bromoacetate (9-2, 9.9 g, 59.2 mmol), KI (1.04 g, 6.26 mmol) and NaHCO.sub.3 (5.0 g, 59.5 mmol). The mixture was stirred at room temperature for 14 h. After completion of the reaction, the resulting mixture was diluted with EtOAc (200 mL), washed with water (200 mL2) and brine (200 mL). The organic layers were dried by anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified by silica gel chromatography (eluted with 20% EtOAc in Petroleum ether) to afford the title compound 9-3 (1.7 g, 81% yield) as a colorless oil.

[0115] Step 2: A mixture of compound 9-3 (500 mg, 1.50 mmol) and TFA (1 mL) in DCM (4 mL) was stirred at room temperature for 1 h. After completion of the reaction, the resulting mixture was concentrated to give the title compound 9-4 (600 mg) as a colorless oil, which was used at the next step without further purification.

[0116] Step 3: Compound 9-4 (600 mg, crude from Step 2) was dissolved in DCM (5 mL) and adjusted pH=8 by adding Et3N. Then the solution was added to a stirring mixture of compound 1-2 (360 mg, 1.24 mmol) and EDCI (431 mg, 2.25 mL) in DCM (10 mL). After stirring for 16 h, the resulting mixture was diluted with DCM (50 mL) and washed with water (50 mL) and 1 M HCl (50 mL2). The organic layer was dried by anhydrous Na.sub.2SO.sub.4, filtered and concentrated to give the title compound 9-5 (600 mg) as a colorless oil, which was used at the next step without further purification.

[0117] Step 4: A mixture of compound 9-5 (600 mg, crude from Step 3) in MeOH (10 mL) was treated with aqueous NaOH (1 M, 3.67 mL, 3.67 mmol), and stirred at room temperature for 2 h. After completion of the reaction, the mixture was concentrated to get rid of the organic solvents. The aqueous solution was washed with EtOAc (50 mL3) and acidified with 1 M HCl to pH=2 to form a white precipitation. The precipitation was filtered and washed with water to give the title compound 9-6 (180 mg, 26% yield over three steps) as a white solid. LCMS: m/z calculated for C.sub.16H.sub.21IN.sub.2O.sub.5: 448.26; found: 449.03 [M+H].sup.+

[0118] Step 5: A mixture of compound 9-6 (180 mg, 0.40 mmol) and compound 9-7 (155 mg, 0.40 mmol) in toluene (18 mL) was stirred at 120 C. for 3 h. After completion of the reaction, the mixture was concentrated. The residue was washed by MTBE and filtered to give a solid. The crude product was re-dissolved in acetonitrile and purified by pre-TLC (SiO.sub.2, DCM/acetonitrile=2/1) to afford the title compound 9 (50.1 mg, 15% yield) as a white solid. .sup.1H NMR (400 MHz, DMSO-d6) 9.09 (s, 1H), 8.90 (d, J=2.5 Hz, 1H), 8.83 (d, J=8.6 Hz, 1H), 8.75 (s, 1H), 8.17 (s, 1H), 7.62 (d, J=7.8 Hz, 2H), 7.42 (d, J=9.7 Hz, 1H), 7.28 (q, J=8.1, 7.4 Hz, 4H), 7.20 (d, J=7.0 Hz, 1H), 7.02 (d, J=7.9 Hz, 2H), 4.74 (q, J=8.2 Hz, 1H), 4.33 (d, J=17.2 Hz, 1H), 4.24-4.11 (m, 2H), 3.79 (d, J=16.8 Hz, 1H), 3.62-3.53 (m, 1H), 3.22 (s, 1H), 3.17 (s, 1H), 3.06 (d, J=8.5 Hz, 2H), 2.51 (s, 1H), 2.17-2.09 (m, 2H), 1.80 (t, J=7.7 Hz, 2H), 1.59 (s, 1H), 1.42 (t, J=12.9 Hz, 1H), 1.25 (d, J=14.1 Hz, 4H), 0.86 (dd, J=13.0, 6.5 Hz, 6H). LCMS: m/z calculated for C.sub.35H.sub.42BIN.sub.6O.sub.7: 796.47; found: 797.6 [M+H].sup.+

Example 10

N-((S)-1-(((R)-1-(6-(4-(4-(4-iodophenyl)butanamido)butyl)-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)-3-methylbutyl)amino)-1-oxo-3-phenylpropan-2-yl)pyrazine-2-carbox amide (10)

##STR00050##

##STR00051##

[0119] Step 1: A solution of compound 1-2 (400 mg, 1.38 mmol), compound 10-1 (312 mg, 1.66 mmol), DMAP (17 mg, 0.14 mmol) and EDCI (397 mg, 2.07 mmol) in DCM (20 mL) was stirred at room temperature for 2 h. After completion of the reaction, the mixture was diluted with DCM (100 mL) and washed with 1N HCl and brine. The organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to give the title compound 10-2 (500 mg), which was used at the next step without further purification.

[0120] Step 2: The compound 10-2 (500 mg, crude from Step 1) was dissolved in 4N HCl in EtOAc (5 mL), and the solution was stirred for 1 h. After completion of the reaction, the mixture was concentrated to give the title compound 10-3 (450 mg), which was used at the next step without further purification.

[0121] Step 3: A mixture of compound 10-3 (450 mg, crude from Step 2), compound 10-4 (600 uL, 4.14 mmol) and triethylamine (1.0 mL, 6.90 mmol) in acetonitrile (10 mL) was stirred at room temperature for 18 h. After completion of the reaction, the reaction mixture was diluted with DCM (100 mL), and washed with water and brine. The organic layer was dried over Na.sub.2SO.sub.4, filtered and concentrated to give the title compound 10-5 (300 mg), which was used at next step without further purification.

[0122] Step 4: The compound 10-5 (300 mg, crude from Step 3) was dissolved in 4 N HCl in EtOAc (5 mL) and stirred for 2 h. After completion of the reaction, the mixture was concentrated. The crude product was purified by prep-HPLC (C18 column, eluted with acetonitrile and H.sub.2O, TFA condition) to give the title compound 10-6 (110 mg, yield: 13.4% over 4 steps) as a light yellow oil.

[0123] Step 5: A mixture of compound 10-6 (112 mg, 0.235 mmol) and compound 9-7 (90 mg, 0.234 mmol) in toluene (5 mL) was stirred at 120 C. for 3 h. After completion of the reaction, the mixture was concentrated. The residue was washed by MTBE and filtered to give a solid. The crude product was re-dissolved in acetonitrile and purified by pre-TLC (SiO.sub.2, DCM/acetonitrile=3/1) to afford the title compound 10 (51 mg, 26% yield) as a white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 9.07 (s, 1H), 8.89 (d, J=8.3 Hz, 1H), 8.86 (d, J=2.3 Hz, 1H), 8.73 (s, 1H), 7.88-7.79 (m, 1H), 7.59 (dd, J=8.2, 1.9 Hz, 2H), 7.34 (d, J=9.6 Hz, 1H), 7.30-7.20 (m, 4H), 7.19-7.12 (m, 1H), 7.03-6.95 (m, 2H), 4.67 (q, J=7.7 Hz, 1H), 4.17 (d, J=17.3 Hz, 1H), 4.05 (d, J=17.4 Hz, 1H), 3.95 (d, J=16.6 Hz, 1H), 3.63-3.45 (m, 2H), 3.07 (d, J=7.3 Hz, 6H), 2.09-2.03 (m, 2H), 1.77 (p, J=7.4 Hz, 2H), 1.56 (s, 3H), 1.41 (t, J=12.6 Hz, 3H), 1.21 (d, J=17.2 Hz, 2H), 0.85 (d, J=6.6 Hz, 3H), 0.80 (d, J=6.3 Hz, 3H). LCMS: m/z calculated for C.sub.37H.sub.46BIN.sub.6O.sub.7: 824.52; found: 825.9. [M+H].sup.+

Example 11

N-((S)-1-(((R)-1-(6-(3-(4-(4-iodophenyl)butanamido)propyl)-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)-3-methylbutyl)amino)-1-oxo-3-phenylpropan-2-yl)pyrazine-2-carbox amide (11)

##STR00052##

##STR00053##

[0124] Step 1: A mixture of compound 11-1 (1.0 g, 6.79 mmol), NaHCO.sub.3 (4.8 g, 57.07 mmol), KI (950 mg, 5.72 mmol) and compound 9-2 (9.5 g, 56.89 mmol) in DMF (20 mL) was stirred at room temperature for 16 h. After completion of the reaction, the mixture was partitioned between EtOAc (100 mL) and water (100 mL). The organic layer was washed with brine (100 mL), dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The crude product was purified by silica gel column (eluted with 15% EtOAc in petroleum ether) to afford the title compound 11-2 (1.7 g, yield 85%) as a colorless oil. TLC (DCM:MeOH/10:1): R.sub.f (compound 11-1)=0.2; TLC (petroleum ether:EtOAc/2:1): R.sub.f (compound 11-2)=0.45.

[0125] Step 2: A solution of compound 11-2 (700 mg, 2.02 mmol) and TFA (4 mL) in DCM (4 mL) was stirred at room temperature for 1 h. After completion of the reaction, the mixture was concentrated to give the title compound 11-3 (680 mg, yield 70%) as a colorless oil, which was used at the next step without further purification. TLC (petroleum ether:EtOAc/2:1): R.sub.f(compound 11-2)=0.45; TLC (DCM:MeOH/5:1): R.sub.f (compound 11-3)=0.15.

[0126] Step 3: A mixture of crude compound 11-3 (680 mg, 1.43 mmol), compound 1-1 (527 mg, 1.82 mmol), EDCI (580 mg, 3.04 mmol) and DMAP (25 mg, 0.205 mmol) in DCM (10 mL) was stirred at room temperature for 16 h. After completion of the reaction, the mixture was partitioned between EtOAc (50 mL) and water (20 mL). The organic layer was washed with 1N HCl (50 mL2), saturated Na.sub.2CO.sub.3 solution, and brine (50 mL), dried over anhydrous Na.sub.2SO.sub.4 and concentrated to afford the title compound 11-4 (580 mg, yield 78%) as a colorless oil, which was used directly at the next step without further purification. LCMS: m/z calculated for C.sub.21H.sub.31IN.sub.2O.sub.5: 518.39; found: 519.09 [M+H].sup.+.

[0127] Step 4: A mixture of compound 11-4 (580 mg, 1.12 mmol) and NaOH (290 mg, 7.25 mmol) in EtOH (10 mL) and H.sub.2O (5 mL) was stirred at room temperature for 4 h. After completion of the reaction, the mixture was concentrated to get rid of the organic solvents. The left aqueous solution was adjusted pH=2 and purified by reverse phase flash chromatography (C18 column, eluted with acetonitrile and water, TFA condition) to afford the title compound 11-5 (220 mg, yield 43%) as white solid. LCMS: m/z calculated for C.sub.17H.sub.23IN.sub.2O.sub.5: 462.28; found: 463.51 [M+H].sup.+.

[0128] Step 5: A mixture of compound 11-5 (120 mg, 0.259 mmol) and compound 9-7 (83 mg, 0.216 mmol) in toluene (5 mL) was stirred at 120 C. for 13 h. After completion of the reaction, the mixture was concentrated. The residue was purified by pre-TLC (DCM:acetonitrile=3:1) to afford 11 (63 mg, yield 35%) as a white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 9.07 (s, 1H), 8.92-8.83 (m, 2H), 8.77-8.70 (m, 1H), 7.74 (s, 1H), 7.63-7.54 (m, 2H), 7.38 (d, J=9.8 Hz, 1H), 7.27 (dd, J=13.3, 7.8 Hz, 4H), 7.19-7.14 (m, 1H), 6.98 (d, J=7.7 Hz, 2H), 4.67 (d, J=8.4 Hz, 1H), 4.17-4.05 (m, 2H), 3.98 (d, J=16.6 Hz, 1H), 3.53 (d, J=22.0 Hz, 2H), 3.07 (d, J=11.9 Hz, 6H), 2.08 (d, J=6.9 Hz, 2H), 1.80-1.70 (m, 3H), 1.57 (s, 2H), 1.42 (t, J=13.1 Hz, 2H), 1.21 (d, J=18.3 Hz, 2H), 0.82 (dd, J=22.8, 6.3 Hz, 6H). LCMS: m/z calculated for C.sub.36H.sub.44BIN.sub.6O.sub.7: 810.50; found: 811.63 [M+H].sup.+.

Example 12

N-((S)-1-(((R)-1-(6-(5-(4-(4-iodophenyl)butanamido)pentyl)-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)-3-methylbutyl)amino)-1-oxo-3-phenylpropan-2-yl)pyrazine-2-carbox amide (12)

##STR00054##

Synthetic Scheme 12

##STR00055##

[0129] Step 1: A mixture of compound 12-1 (500 mg, 2.47 mmol), NaHCO.sub.3 (2.1 g, 24.9 mmol), KI (410 mg, 2.47 mmol) and compound 9-2 (4.1 g, 24.5 mmol) in DMF (10 mL) was stirred at room temperature for 16 h. After completion of the reaction, the mixture was partitioned between EtOAc (50 mL) and water (50 mL). The organic layer was washed with brine (50 mL), dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The crude product was purified by silica gel column (eluted with 15% EtOAc in petroleum ether) to afford the title compound 12-2 (700 mg, yield 75%) as colorless oil. TLC (DCM:MeOH/10:1): R.sub.f (compound 12-1)=0.2; TLC (petroleum ether:EtOAc/2:1): R.sub.f (compound 12-2)=0.55.

[0130] Step 2: A solution of compound 12-2 (700 mg, 1.87 mmol) and TFA (4 mL) in DCM (4 mL) was stirred at room temperature for 1 h. After completion of the reaction, the mixture was concentrated to give the title compound 12-3 (760 mg, yield 80%) as a colorless oil, which was used at the next step without further purification. TLC (petroleum ether:EtOAc/2:1): R.sub.f(compound 12-2)=0.55; TLC (DCM:MeOH/5:1): R.sub.f (compound 12-3)=0.18.

[0131] Step 3: A mixture of crude compound 12-3 (660 mg, 1.51 mmol), compound 1-2 (488 mg, 1.68 mmol), EDCI (540 mg, 2.82 mmol) and DMAP (23 mg, 0.189 mmol) in DCM (10 mL) was stirred at room temperature for 16 h. After completion of the reaction, the mixture was partitioned between EtOAc (50 mL) and water (20 mL). The organic layer was washed with 1N HCl (50 mL2), saturated Na.sub.2CO.sub.3 solution and brine (50 mL), dried over anhydrous Na.sub.2SO.sub.4 and concentrated to afford the title compound 12-4 (700 mg, yield 84%) as a colorless oil, which was used at the next step without further purification. TLC (petroleum ether:EtOAc/2:1): R.sub.f(compound 12-3)=0.5; R.sub.f (compound 12-4)=0.2.

[0132] Step 4: A mixture of compound 12-4 (700 mg, 1.28 mmol) and NaOH (270 mg, 6.75 mmol) in EtOH (10 mL) and H.sub.2O (5 mL) was stirred at room temperature for 4 h. After completion of the reaction, the mixture was concentrated to get rid of the organic solvents. The left aqueous solution was adjusted to pH=2 and purified directly by reverse phase flash chromatography (C18 column, eluted with acetonitrile and water, TFA condition) to afford the title compound 12-5 (270 mg, yield 43%) as white solid. LCMS: m/z calculated for C.sub.19H.sub.27IN.sub.2O.sub.5: 490.34; found: 491.30 [M+H].sup.+.

[0133] Step 5: A mixture of compound 12-5 (153 mg, 0.312 mmol) and compound 9-7 (100 mg, 0.260 mmol) in toluene (5 mL) was stirred at 120 C. for 13 h. After completion of the reaction, the mixture was concentrated. The residue was purified by pre-TLC (DCM:acetonitrile=3:1) to afford 12 (67 mg, yield 25%) as a white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 9.05 (d, J=3.0 Hz, 1H), 8.94-8.82 (m, 2H), 8.71 (d, J=3.3 Hz, 1H), 7.81 (s, 1H), 7.56 (dd, J=8.1, 2.8 Hz, 2H), 7.35 (d, J=10.1 Hz, 2H), 7.30-7.21 (m, 4H), 7.19-7.13 (m, 1H), 6.96 (dd, J=8.4, 2.8 Hz, 2H), 4.66 (s, 1H), 4.15 (dd, J=17.5, 3.0 Hz, 1H), 4.07-3.99 (m, 1H), 3.93 (dd, J=16.8, 3.0 Hz, 1H), 3.05 (d, J=8.4 Hz, 6H), 2.03 (d, J=8.1 Hz, 2H), 1.75 (d, J=8.0 Hz, 2H), 1.54 (s, 4H), 1.41 (d, J=9.9 Hz, 3H), 1.15 (s, 4H), 0.83 (dd, J=6.6, 3.1 Hz, 3H), 0.79 (dd, J=6.6, 3.1 Hz, 3H). LCMS: m/z calculated for C.sub.38H.sub.48BIN.sub.6O.sub.7: 838.55; found: 839.50 [M+H].sup.+.

Example 13

N-((2S)-1-(((1R)-1-(6-(4-(2-(4-isobutylphenyl)propanamido)butyl)-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)-3-methylbutyl)amino)-1-oxo-3-phenylpropan-2-yl)pyrazine-2-carboxamide (13)

##STR00056##

##STR00057##

[0134] Step 1: A mixture of compound 10-1 (2 g, 10.62 mmol), compound 9-2 (17.7 g, 106.23 mmol), NaHCO.sub.3 (8.92 g, 106.23 mmol) and KI (1.76 g, 10.62 mmol) in DMF (20 mL) was stirred at room temperature for 16 h. The reaction mixture was diluted with EtOAc (100 mL) and washed with water (100 mL). The organic layer was washed with brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The crude product was purified by silica gel column (eluted with 30% EtOAc in Petroleum ether) to afford the title compound 13-1 (3.5 g, yield 91%) as a yellow oil.

[0135] Step 2: To a solution of compound 13-1 (2 g, 5.55 mmol) in DCM (16 mL) was added TFA (4 mL) at room temperature and the mixture stirred for 1 h. After completion of the reaction, the mixture was concentrated to afford the crude compound 13-2 (3 g), which was used directly at the next step.

[0136] Step 3: A mixture of compound 5-1 (1.14 g, 5.53 mmol), HATU (3.15 g, 8.29 mmol) and DIPEA (1.37 mL, 8.29 mmol) in DMF (2 mL) was stirred at room temperature for 10 minutes. Compound 13-2 (3 g, crude from Step 2) was added and the mixture was stirred for 3 h. After completion of the reaction, the mixture was diluted with EtOAc (50 mL) and washed with water (50 mL). The organic layer was washed with brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The crude product was purified by reverse phase flash chromatography (C18 column, eluted with acetonitrile and H.sub.2O, HCl condition) to afford the title compound 13-3 (1.8 g, yield 72%, 2 steps). LCMS: m/z calculated for C.sub.25H.sub.40N.sub.2O.sub.5: 448.29; found: 473.32 [M+H].sup.+.

[0137] Step 4: To a mixture of compound 13-3 (1.8 g, 4.01 mmol) in EtOH (8 mL) and H.sub.2O (4 mL) was added LiOH (336.7 mg, 20.06 mmol). The mixture was stirred at room temperature for 1 h. After completion of the reaction, the mixture was adjusted to pH=3 with 1 N HCl and concentrated. The crude product was purified by reverse phase flash chromatography (C18 column, eluted with acetonitrile and H.sub.2O, HCl condition) to afford the title compound 13-4 (1.1 g, yield 70%). LCMS: m/z calculated for C.sub.21H.sub.32N.sub.2O.sub.5: 392.50; found: 393.33 [M+H].sup.+.

[0138] Step 5: A mixture of compound 13-4 (1 g, 0.36 mmol) and compound 9-7 (115 mg, 0.30 mmol) in toluene (15 mL) was stirred at 120 C. for 17 h. After completion of the reaction, the mixture was concentrated. The crude product was purified by silica gel column (eluted with 30% acetonitrile in DCM) to afford the title compound 13 (1 g, yield 53%) as a pale-yellow solid. .sup.1H NMR (500 MHz, DMSO-d.sub.6) 9.09 (dd, J=3.8, 1.5 Hz, 1H), 8.92-8.85 (m, 2H), 8.73 (ddd, J=5.1, 2.5, 1.5 Hz, 1H), 7.97 (td, J=5.8, 2.1 Hz, 1H), 7.35 (dd, J=9.9, 6.0 Hz, 1H), 7.31-7.15 (m, 7H), 7.06 (d, J=7.9 Hz, 2H), 4.73-4.63 (m, 1H), 4.12 (dd, J=17.4, 3.6 Hz, 1H), 4.03 (dd, J=17.3, 1.7 Hz, 1H), 3.98-3.89 (m, 1H), 3.56 (qd, J=8.1, 7.0, 3.5 Hz, 2H), 3.48 (dd, J=16.7, 8.7 Hz, 1H), 3.15-2.97 (m, 6H), 2.38 (d, J=7.2 Hz, 2H), 1.78 (dp, J=13.5, 6.7 Hz, 1H), 1.62-1.50 (m, 2H), 1.50-1.40 (m, 2H), 1.40-1.33 (m, 2H), 1.32 (dd, J=7.0, 1.4 Hz, 3H), 1.21 (ddt, J=13.7, 10.2, 4.5 Hz, 1H), 0.92-0.74 (m, 12H). LCMS: m/z calculated for C.sub.40H.sub.53BN.sub.6O.sub.7: 740.41; found: 742.01 [M+H].sup.+.

Example 14

N-((S)-1-(((R)-1-(6-(4-((S)-2-(6-methoxynaphthalen-2-yl)propanamido)butyl)-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)-3-methylbutyl)amino)-1-oxo-3-phenylpropan-2-yl) pyrazine-2-carboxamide (14)

##STR00058##

##STR00059##

[0139] Step 1: A mixture of compound 6-1 (254 mg, 1.11 mmol), DIPEA (445 mg, 3.45 mmol), compound 13-2 (415 mg, crude, 70% purity, 1.12 mmol) and HATU (787 mg, 2.07 mmol) in DMF (3 mL) was stirred at room temperature for 3 h. After completion of the reaction, the mixture was partitioned between EtOAc (50 mL) and water (50 mL). The organic layer was washed with brine, dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The crude product was purified by reverse phase flash chromatography (C18 column, eluted with acetonitrile and water, HCl condition). The desired component was lyophilized to afford the title compound 14-1 (380 mg, yield 58%). LCMS: m/z calculated for C.sub.26H.sub.36N.sub.2O.sub.6: 472.3; found: 473.32 [M+H].sup.+.

[0140] Step 2: A mixture of compound 14-1 (380 mg, 0.80 mmol) and NaOH (160 mg, 4.01 mmol) in EtOH (5 mL) and H.sub.2O (5 mL) was stirred at room temperature for 3 h. After completion of the reaction, the mixture was adjusted to pH=7 with 1 N HCl. The resulting mixture was concentrated and purified by reverse phase flash chromatography (C18 column, eluted with acetonitrile and water, HCl condition). The desired component was lyophilized to afford the title compound 14-2 (150 mg, yield 45%). LCMS: m/z calculated for C.sub.26H.sub.36N.sub.2O.sub.6: 416.19; found: 417.13. [M+H].sup.+

[0141] Step 3: A mixture of compound 14-2 (150 mg, 0.36 mmol) and compound 9-7 (115 mg, 0.30 mmol) in toluene (3 mL) was stirred at 120 C. for 17 h. After completion of the reaction, the mixture was concentrated to get rid of organic solvents. The crude product was purified by pre-HPLC (C18 column, eluted with acetonitrile and H.sub.2O, neutral condition) to afford the title compound 14 (30 mg, yield 13%). .sup.1H NMR (400 MHz, DMSO-d.sub.6) 9.05 (d, J=4.0 Hz, 1H), 8.86 (d, J=13.7 Hz, 2H), 8.68 (s, 1H), 8.05 (d, J=6.4 Hz, 1H), 7.72 (dt, J=13.6, 6.0 Hz, 3H), 7.47-7.40 (m, 1H), 7.34 (d, J=9.1 Hz, 1H), 7.28-7.19 (m, 5H), 7.18-7.08 (m, 2H), 4.66 (s, 1H), 4.09 (d, J=17.7 Hz, 1H), 3.98 (d, J=17.5 Hz, 1H), 3.88 (d, J=14.9 Hz, 1H), 3.83 (d, J=4.3 Hz, 3H), 3.71 (s, 2H), 3.06 (s, 6H), 1.53 (s, 2H), 1.39 (q, J=13.4, 9.6 Hz, 6H), 1.28-1.12 (m, 4H), 0.83 (t, J=5.2 Hz, 3H), 0.79 (t, J=5.5 Hz, 3H). LCMS: m/z calculated for C.sub.41H.sub.49BN.sub.6O.sub.8: 764.37; found: 765.43. [M+H].sup.+.

Example 15

N-((S)-1-(((R)-1-(6-(4-((R)-2-(4-isobutylphenyl)propanamido)butyl)-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)-3-methylbutyl)amino)-1-oxo-3-phenylpropan-2-yl)pyrazine-2-carboxamide (15)

##STR00060##

##STR00061##

[0142] Step 1: A mixture of compound 7-1 (143 mg, 0.69 mmol), HATU (396 mg, 1.04 mmol) and DIPEA (224 mg, 1.73 mmol) in DMF (2 mL) was stirred at room temperature for 10 minutes, compound 13-2 (300 mg, crude, 70% purity, 0.81 mmol) was added. After stirring at room temperature for 3 h, the mixture was diluted with EtOAc (20 mL) and washed with water (20 mL). The organic layer was washed with brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The crude product was purified by reverse phase flash chromatography (C18 column, eluted with acetonitrile and H.sub.2O, HCl condition) to afford the title compound 15-1 (307 mg, yield 99%). LCMS: m/z calculated for C.sub.25H.sub.40N.sub.2O.sub.5: 448.60; found: 449.62 [M+H].sup.+.

[0143] Step 2: To a mixture of compound 15-1 (337 mg, 0.75 mmol) in EtOH (2 mL) and H.sub.2O (2 mL) was added LiOH.Math.H.sub.2O (157.6 mg, 3.76 mmol). Then the mixture was stirred at room temperature for 1 h. After completion of the reaction, the mixture was adjusted to pH=7 with 1 N HCl and concentrated. The crude product was purified by reverse phase flash chromatography (C18 column, eluted with acetonitrile and H.sub.2O, HCl condition) to afford the title compound 15-2 (231 mg, yield 78%). LCMS: m/z calculated for C.sub.25H.sub.40N.sub.2O.sub.5: 392.23; found: 393.33 [M+H].sup.+.

[0144] Step 3: A mixture of compound 15-2 (100 mg, 0.25 mmol) and compound 9-7 (81.6 mg, 0.21 mmol) in toluene (5 mL) was stirred at 120 C. for 17 h. After completion of the reaction, the mixture was concentrated. The crude product was purified by pre-TLC (SiO.sub.2, acetonitrile/DCM=1/3) to afford the title compound 15 (97 mg, yield 61%) as a white solid. .sup.1H NMR (500 MHz, DMSO-d.sub.6) 9.09 (d, J=1.5 Hz, 1H), 8.92-8.86 (m, 2H), 8.74 (dd, J=2.5, 1.5 Hz, 1H), 7.97 (t, J=5.8 Hz, 1H), 7.34 (d, J=9.8 Hz, 1H), 7.31-7.27 (m, 2H), 7.27-7.23 (m, 2H), 7.22-7.19 (m, 2H), 7.19-7.15 (m, 1H), 7.08-7.03 (m, 2H), 4.68 (q, J=7.7 Hz, 1H), 4.11 (d, J=17.4 Hz, 1H), 4.03 (d, J=17.3 Hz, 1H), 3.92 (d, J=16.7 Hz, 1H), 3.56 (qd, J=6.9, 3.1 Hz, 2H), 3.49 (d, J=16.8 Hz, 1H), 3.13-2.98 (m, 6H), 2.38 (d, J=7.1 Hz, 2H), 1.78 (hept, J=6.8 Hz, 1H), 1.57 (ddt, J=13.7, 6.7, 3.6 Hz, 2H), 1.49-1.34 (m, 4H), 1.32 (d, J=7.1 Hz, 3H), 1.20 (ddd, J=17.2, 10.2, 5.0 Hz, 1H), 0.83 (dt, J=13.6, 6.9 Hz, 12H). LCMS: m/z calculated for C.sub.40H.sub.53BN.sub.6O.sub.7: 740.41; found: 741.94 [M+H].sup.+.

Example 16

N-((S)-1-(((R)-1-(6-(4-((S)-2-(4-isobutylphenyl)propanamido)butyl)-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)-3-methylbutyl)amino)-1-oxo-3-phenylpropan-2-yl)pyrazine-2-carboxamide (16)

##STR00062##

##STR00063##

[0145] Step 1: A mixture of compound 8-1 (334 mg, 1.62 mmol), HATU (924 mg, 2.42 mmol) and DIPEA (716.8 L, 4.04 mmol) in DMF (5 mL) was stirred at room temperature for 10 minutes, compound 13-2 (800 mg, crude, 70% purity, 2.15 mmol) was added. After stirring at room temperature for 3 h, the mixture was diluted with EtOAc (50 mL) and washed with water (50 mL). The organic layer was washed with brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The crude product was purified by reverse phase flash chromatography (C18 column, eluted with acetonitrile and H.sub.2O, HCl condition) to afford the title compound 16-1 (706 mg, yield 97%). LCMS: m/z calculated for C.sub.25H.sub.40N.sub.2O.sub.5: 448.60; found: 449.62 [M+H].sup.+.

[0146] Step 2: To a mixture of compound 16-1 (806 mg, 1.80 mmol) in EtOH (5 mL) and H.sub.2O (5 mL) was added LiOH.Math.H.sub.2O (376.9 mg, 8.98 mmol), and the mixture was stirred at room temperature for 1 h. After completion of the reaction, the mixture was adjusted to pH=7 with 1 N HCl and concentrated. The crude product was purified by reverse phase flash chromatography (C18 column, eluted with acetonitrile and H.sub.2O, HCl condition) to afford the title compound 16-2 (549 mg, yield 78%). LCMS: m/z calculated for C.sub.25H.sub.40N.sub.2O.sub.5: 392.23; found: 393.33 [M+H].sup.+.

[0147] Step 3: A mixture of compound 16-2 (100 mg, 0.25 mmol) and compound 9-7 (81.6 mg, 0.21 mmol) in toluene (5 mL) was stirred at 120 C. for 17 h. After completion of the reaction, the mixture was concentrated. The crude product was purified by pre-TLC (Acetonitrile/DCM=1/3) to afford the compound 16 (98 mg, yield 62%) as a white solid. .sup.1H NMR (500 MHz, DMSO-d.sub.6) 9.09 (d, J=1.5 Hz, 1H), 8.90-8.85 (m, 2H), 8.74-8.71 (m, 1H), 7.97 (t, J=5.8 Hz, 1H), 7.36 (d, J=9.8 Hz, 1H), 7.31-7.27 (m, 2H), 7.27-7.23 (m, 2H), 7.23-7.20 (m, 2H), 7.19-7.15 (m, 1H), 7.08-7.04 (m, 2H), 4.75-4.64 (m, 1H), 4.12 (d, J=17.4 Hz, 1H), 4.03 (d, J=17.3 Hz, 1H), 3.92 (d, J=16.7 Hz, 1H), 3.61-3.52 (m, 2H), 3.47 (d, J=16.7 Hz, 1H), 3.14-2.98 (m, 6H), 2.38 (d, J=7.1 Hz, 2H), 1.79 (dh, J=13.6, 6.8 Hz, 1H), 1.62-1.50 (m, 2H), 1.50-1.34 (m, 4H), 1.32 (d, J=7.0 Hz, 3H), 1.21 (ddd, J=17.2, 9.5, 4.3 Hz, 1H), 0.84 (td, J=10.4, 9.7, 6.5 Hz, 12H). LCMS: m/z calculated for C.sub.40H.sub.53BN.sub.6O.sub.7: 740.41; found: 741.73 [M+H].sup.+.

Example 17

N-((S)-1-(((R)-1-(6-(5-((S)-2-(4-isobutylphenyl)propanamido)pentyl)-4,8-dioxo-1,3,6,2-dioxa zaborocan-2-yl)-3-methylbutyl)amino)-1-oxo-3-phenylpropan-2-yl)pyrazine-2-carboxamide (17)

##STR00064##

##STR00065##

[0148] Step 1: To a solution of compound 17-1 (1.0 g, 4.94 mmol) in DMF (10 mL) was added NaHCO.sub.3 (4.15 g, 49.39 mmol), KI (820 mg, 4.93 mmol) and compound 9-2 (4.13 g, 24.73 mmol). The mixture was stirred at room temperature for 16 h. After completion of the reaction, the mixture was partitioned between EtOAc (100 mL) and water (100 mL). The organic layer was washed with brine, dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The residue was purified by silica gel column (eluted with 30% EtOAc in petroleum ether) to afford the title compound 17-2 (1.66 g, yield 89%).

[0149] Step 2: A mixture of compound 17-2 (1 g, 2.67 mmol) in 4 N HCl (EtOAc solution, 10 mL) and MeOH (0.5 mL) was stirred at room temperature for 2 h. After completion of the reaction, the mixture was concentrated to afford the title compound 17-3 (1 g, crude), which was used at the next step without further purification.

[0150] Step 3: A solution of compound 17-3 (200 mg, 0.673 mmol), compound 8-1 (660 mg, 3.20 mmol), DIPEA (862 mg, 6.68 mmol) and HATU (1.52 g, 3.99 mmol) in DMF (5 mL) was stirred at room temperature for 3 h. After completion of the reaction, the mixture was diluted with EtOAc (50 mL) and washed with water (50 mL2) and brine (50 mL). The organic layer was dried by anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The crude product was purified by silica gel column (eluted with 30% EtOAc in petroleum ether) to afford the title compound 17-4 (623 mg, yield 50% over 2 steps). LCMS: m/z calculated for C.sub.26H.sub.42N.sub.2O.sub.5: 462.63; found: 463.95 [M+H].sup.+.

[0151] Step 4: To a solution of compound 17-4 (620 mg, 1.34 mmol) in EtOH (6 mL) and H.sub.2O (3 mL) was added NaOH (214 mg, 5.36 mmol) at 0 C. and the reaction was stirred at room temperature for 1 h. The mixture was concentrated and adjusted to pH=2 with 1 N HCl at 0 C. The aqueous mixture was purified by reverse phase flash chromatography (C18 column, eluted with 55% acetonitrile in water, HCl condition). The desired components were lyophilized to afford the title compound 17-5 (209 mg, yield 35%) as a white solid. LCMS: m/z calculated for C.sub.22H.sub.34N.sub.2O.sub.5: 406.52; found: 407.83 [M+H].sup.+.

[0152] Step 5: A mixture of compound 17-5 (172 mg, 0.388 mmol) and compound 9-7 (149 mg, 0.388 mmol) in toluene (3 mL) was stirred at 120 C. for 17 h. After completion of the reaction, the mixture was concentrated. The crude product was purified by pre-TLC (acetonitrile:dichloromethane=1:3) to afford 17 (55 mg, yield 18%) as a white solid. LCMS: m/z calculated for C.sub.41H.sub.55BN.sub.6O.sub.7: 754.74; found: 755.93 [M+H].sup.+.

Example 18

N-((S)-1-(((R)-1-(6-(6-((S)-2-(4-isobutylphenyl)propanamido)hexyl)-4,8-dioxo-1,3,6,2-dioxaz aborocan-2-yl)-3-methylbutyl)amino)-1-oxo-3-phenylpropan-2-yl)pyrazine-2-Carboxamide (18)

##STR00066##

##STR00067##

[0153] Step 1: To a solution of compound 18-1 (1.0 g, 4.62 mmol) in DMF (10 mL) was added NaHCO.sub.3 (3.88 g, 46.18 mmol), KI (767 mg, 4.59 mmol) and compound 9-2 (3.86 g, 23.11 mmol). The mixture was stirred at room temperature for 16 h. After completion of the reaction, the mixture was partitioned between EtOAc (100 mL) and water (100 mL). The organic layer was washed with brine, dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The residue was purified by silica gel column (eluted with 30% EtOAc in petroleum ether) to afford the title compound 18-2 (1.24 g, yield 69%).

[0154] Step 2: A mixture of compound 18-2 (700 mg, 1.80 mmol) in 4 N HCl (EtOAc solution, 7 mL) and MeOH (0.5 mL) was stirred at room temperature for 2 h. After completion of the reaction, the mixture was concentrated to afford the title compound 18-3 (622 mg, crude), which was used at the next step without further purification.

[0155] Step 3: A solution of compound 18-3 (622 mg, crude), compound 8-1 (371 mg, 1.79 mmol), DIPEA (581 mg, 4.50 mmol) and HATU (1.03 g, 2.70 mmol) in DMF (5 mL) was stirred at room temperature for 3 h. After completion of the reaction, the mixture was diluted with EtOAc (50 mL) and washed with water (50 mL2) and brine (50 mL). The organic layer was dried by anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The crude product was purified by silica gel column (eluted with 30% EtOAc in petroleum ether) to afford the title compound 18-4 (403 mg, yield 46% over 2 steps). LCMS: m/z calculated for C.sub.27H.sub.44N.sub.2O.sub.5: 476.66; found: 478.01 [M+H].sup.+.

[0156] Step 4: To a solution of compound 18-4 (403 mg, 0.845 mmol) in EtOH (4 mL) and H.sub.2O (2 mL) was added NaOH (135 mg, 3.37 mmol) at 0 C. and the reaction was stirred at room temperature for 1 h. The mixture was concentrated and adjusted to pH=2 with 1 N HCl at 0 C. The aqueous mixture was purified by reverse phase flash chromatography (C18 column, eluted with 50% acetonitrile in water, HCl condition). The desired components were lyophilized to afford the title compound 18-5 (181 mg, yield 46%) as a white solid. LCMS: m/z calculated for C.sub.23H.sub.36N.sub.2O.sub.5: 420.55; found: 421.76 [M+H].sup.+.

[0157] Step 5: A mixture of compound 18-5 (178 mg, 0.389 mmol) and compound 9-7 (149 mg, 0.388 mmol) in toluene (3 mL) was stirred at 120 C. for 16 h. After completion of the reaction, the mixture was concentrated. The crude product was purified by pre-TLC (acetonitrile:dichloromethane=1:3) to afford 18 (65 mg, yield 21%) as a white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 9.07 (d, J=1.4 Hz, 1H), 8.90 (d, J=8.4 Hz, 1H), 8.86 (d, J=2.5 Hz, 1H), 8.72 (dd, J=2.5, 1.5 Hz, 1H), 7.87 (t, J=5.5 Hz, 1H), 7.32 (d, J=9.8 Hz, 1H), 7.29-7.22 (m, 4H), 7.21-7.13 (m, 3H), 7.05 (d, J=8.0 Hz, 2H), 4.67 (td, J=8.4, 6.4 Hz, 1H), 4.16 (d, J=17.4 Hz, 1H), 4.04 (d, J=17.4 Hz, 1H), 3.94 (d, J=16.7 Hz, 1H), 3.60-3.50 (m, 2H), 3.46 (d, J=16.7 Hz, 1H), 3.09-2.95 (m, 6H), 2.38 (d, J=7.1 Hz, 2H), 1.77 (dt, J=13.5, 6.7 Hz, 1H), 1.61-1.51 (m, 2H), 1.42-1.33 (m, 3H), 1.29 (d, J=7.1 Hz, 4H), 1.26-1.15 (m, 5H), 0.83 (td, J=8.3, 6.4 Hz, 12H). LCMS: m/z calculated for C.sub.42H.sub.57BN.sub.6O.sub.7: 768.76; found: 769.93 [M+H].sup.+.

Example 19

N-((S)-1-(((R)-1-(6-(7-((S)-2-(4-isobutylphenyl)propanamido)heptyl)-4,8-dioxo-1,3,6,2-dioxa zaborocan-2-yl)-3-methylbutyl)amino)-1-oxo-3-phenylpropan-2-yl)pyrazine-2-carboxamide (19)

##STR00068##

##STR00069##

[0158] Step 1: A mixture of compound 8-1 (1 g, 4.84 mmol), NHS (7.27 mmol), DMAP (60 mg, 0.491 mmol) and EDCI (1.86 g, 9.73 mmol) in DCM (10 mL) was stirred at room temperature for 16 h. After completion of the reaction, the mixture was diluted with DCM (50 mL) and washed with 1 N HCl (50 mL) and water (50 mL). The organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to afford the title compound 19-1 (1.26 g, yield 85%) as a white solid.

[0159] Step 2: To a solution of compound 19-2 (996 mg, 7.66 mmol) and DIPEA (987 mg, 7.65 mmol) in DCM (5 mL) was added a solution of compound 19-1 (1.16 g, 3.82 mmol) in DCM (6 mL) over 1 h. After the reaction was stirred at room temperature for 4 h, the mixture was concentrated. The crude product was purified by reverse phase flash chromatography (C18 column, eluted with 80% MeOH in water, HCl condition). The desired components were concentrated to afford the title compound 19-3 (HCl salt, 667 mg, yield 49%) as a colorless oil. LCMS: m/z calculated for C.sub.20H.sub.34N.sub.2O: 318.51; found: 319.88 [M+H].sup.+.

[0160] Step 3: To a solution of compound 19-3 (665 mg, 2.08 mmol) in DMF (8 mL) was added NaHCO.sub.3 (1.75 g, 20.83 mmol), KI (346 mg, 2.08 mmol) and compound 9-2 (1.74 g, 10.41 mmol). The mixture was stirred at room temperature for 16 h. After completion of the reaction, the mixture was partitioned between EtOAc (50 mL) and water (50 mL). The organic layer was washed with brine, dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The residue was purified by silica gel column (eluted with 50% EtOAc in petroleum ether) to afford the title compound 19-4 (629 mg, yield 61%). LCMS: m/z calculated for C.sub.28H.sub.46N.sub.2O.sub.5: 490.69; found: 491.94 [M+H].sup.+.

[0161] Step 4: To a solution of compound 19-4 (627 mg, 1.27 mmol) in EtOH (6 mL) and H.sub.2O (3 mL) was added NaOH (204 mg, 5.1 mmol) at 0 C. and the reaction was stirred at room temperature for 1 h. The mixture was concentrated and adjusted to pH=2 with 1 N HCl at 0 C. The aqueous mixture was purified by reverse phase flash chromatography (C18 column, eluted with 55% acetonitrile in water, HCl condition). The desired components were lyophilized to afford the title compound 19-5 (229 mg, yield 38%) as a white solid. LCMS: m/z calculated for C.sub.24H.sub.38N.sub.2O.sub.5: 434.58; found: 435.75 [M+H].sup.+.

[0162] Step 5: A mixture of compound 19-5 (195 mg, 0.414 mmol) and compound 9-7 (159 mg, 0.414 mmol) in toluene (5 mL) was stirred at 120 C. for 17 h. After completion of the reaction, the mixture was concentrated. The crude product was purified by pre-TLC (acetonitrile:dichloromethane=1:3) to afford 19 (170 mg, yield 52%) as a white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 9.07 (d, J=1.5 Hz, 1H), 8.90 (d, J=8.4 Hz, 1H), 8.86 (d, J=2.5 Hz, 1H), 8.72 (q, J=1.8 Hz, 1H), 7.86 (t, J=5.6 Hz, 1H), 7.35-7.25 (m, 4H), 7.24-7.20 (m, 2H), 7.17 (d, J=10.7 Hz, 2H), 7.05 (dd, J=8.2, 2.0 Hz, 2H), 4.68 (td, J=8.3, 5.6 Hz, 1H), 4.17 (d, J=17.4 Hz, 1H), 4.04 (d, J=17.4 Hz, 1H), 3.95 (d, J=16.6 Hz, 1H), 3.60-3.50 (m, 2H), 3.47 (d, J=17.0 Hz, 1H), 3.10-2.97 (m, 6H), 2.37 (dd, J=7.2, 2.5 Hz, 2H), 1.78 (ddd, J=13.2, 6.6, 2.2 Hz, 1H), 1.56 (dp, J=10.3, 3.4 Hz, 2H), 1.42-1.33 (m, 3H), 1.31-1.11 (m, 11H), 0.86-0.80 (m, 12H). LCMS: m/z calculated for C.sub.43H.sub.59BN.sub.6O.sub.7: 782.79; found: 783.86 [M+H].sup.+.

Example 20

N-((S)-1-(((R)-1-(6-(8-((S)-2-(4-isobutylphenyl)propanamido)octyl)-4,8-dioxo-1,3,6,2-dioxaz aborocan-2-yl)-3-methylbutyl)amino)-1-oxo-3-phenylpropan-2-yl)pyrazine-2-carboxamide (20)

##STR00070##

##STR00071##

[0163] Step 1: To a solution of compound 20-1 (1.0 g, 4.09 mmol) in DMF (10 mL) was added NaHCO.sub.3 (3.44 g, 40.94 mmol), KI (679 mg, 4.09 mmol) and compound 9-2 (3.42 g, 20.47 mmol). The mixture was stirred at room temperature for 16 h. After completion of the reaction, the mixture was partitioned between EtOAc (100 mL) and water (100 mL). The organic layer was washed with brine, dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The residue was purified by silica gel column (eluted with 30% EtOAc in petroleum ether) to afford the title compound 20-2 (1.25 g, yield 69%).

[0164] Step 2: A mixture of compound 20-2 (700 mg, 1.68 mmol) in 4 N HCl (EtOAc solution, 7 mL) and MeOH (0.5 mL) was stirred at room temperature for 2 h. After completion of the reaction, the mixture was concentrated to afford the title compound 20-3 (637 mg, crude), which was used at next step without further purification.

[0165] Step 3: A solution of compound 20-3 (637 mg, crude from Step 2), compound 8-1 (346 mg, 1.67 mmol), DIPEA (542 mg, 4.20 mmol) and HATU (958 mg, 2.52 mmol) in DMF (5 mL) was stirred at room temperature for 16 h. After completion of the reaction, the mixture was diluted with EtOAc (50 mL) and washed with water (50 mL2) and brine (50 mL). The organic layer was dried by anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The crude product was purified by silica gel column (eluted with 30% EtOAc in petroleum ether) to afford the title compound 20-4 (362 mg, yield 42% over 2 steps). LCMS: m/z calculated for C.sub.29H.sub.48N.sub.2O.sub.5: 504.71; found: 506.07 [M+H].sup.+.

[0166] Step 4: To a solution of compound 20-4 (360 mg, 0.713 mmol) in EtOH (4 mL) and H.sub.2O (2 mL) was added NaOH (114 mg, 2.85 mmol) at 0 C. and the reaction was stirred at room temperature for 1 h. The mixture was concentrated and adjusted to pH=2 with 1 N HCl at 0 C. The aqueous mixture was purified by reverse phase flash chromatography (C18 column, eluted with 50% acetonitrile in water, HCl condition). The desired components were lyophilized to afford the title compound 20-5 (247 mg, yield 71%) as a white solid. LCMS: m/z calculated for C.sub.25H.sub.40N.sub.2O.sub.5: 448.60; found: 449.95 [M+H].sup.+.

[0167] Step 5: A mixture of compound 20-5 (178 mg, 0.367 mmol) and compound 9-7 (141 mg, 0.366 mmol) in toluene (3 mL) was stirred at 120 C. for 16 h. After completion of the reaction, the mixture was concentrated. The crude product was purified by pre-TLC (acetonitrile:dichloromethane=1:3) to afford 20 (50 mg, yield 17%) as a white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 9.07 (d, J=1.5 Hz, 1H), 8.90 (d, J=8.5 Hz, 1H), 8.86 (d, J=2.5 Hz, 1H), 8.73 (t, J=1.9 Hz, 1H), 7.85 (t, J=5.6 Hz, 1H), 7.32 (d, J=9.7 Hz, 1H), 7.29-7.23 (m, 4H), 7.23-7.17 (m, 3H), 7.05 (d, J=7.8 Hz, 2H), 4.68 (td, J=8.7, 5.8 Hz, 1H), 4.17 (d, J=17.4 Hz, 1H), 4.04 (d, J=17.4 Hz, 1H), 3.95 (d, J=16.6 Hz, 1H), 3.60-3.51 (m, 2H), 3.48 (d, J=16.6 Hz, 1H), 3.08-2.97 (m, 6H), 2.38 (d, J=7.1 Hz, 2H), 1.78 (dt, J=13.5, 6.7 Hz, 1H), 1.61-1.52 (m, 2H), 1.37 (dd, J=21.4, 4.9 Hz, 3H), 1.28 (d, J=7.1 Hz, 4H), 1.26-1.17 (m, 9H), 0.83 (q, J=8.5, 7.5 Hz, 12H). LCMS: m/z calculated for C.sub.44H.sub.61BN.sub.6O.sub.7: 796.82; found: 797.99[M+H].sup.+.

Example 21

N-((S)-1-(((R)-1-(6-(4-(((S)-2-(4-isobutylphenyl)propanamido)methyl)cyclohexyl)-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)-3-methylbutyl)amino)-1-oxo-3-phenylpropan-2-yl)pyrazine-2-carboxamide (21)

##STR00072##

##STR00073##

[0168] Step 1: A mixture of compound 21-1 (700 mg, 3.07 mmol), 10 w % Pd/C (350 mg) and ammonium hydroxide (28%, 9.5 mL) in MeOH (10 mL) was purged with hydrogen 3 times and stirred under hydrogen atmosphere at room temperature for 16 h. The mixture was concentrated to afford the title compound 21-2 (746 mg, crude) as a colorless oil, which was used at the next step without further purification.

[0169] Step 2: To a solution of compound 21-2 (380 mg, 1.66 mmol) in DMF (5 mL) was added NaHCO.sub.3 (1.39 g, 16.54 mmol), KI (275 mg, 1.65 mmol) and compound 9-2 (1.38 g, 8.26 mmol). The mixture was stirred at room temperature for 16 h. After completion of the reaction, the mixture was partitioned between EtOAc (50 mL) and water (50 mL). The organic layer was washed with brine, dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The residue was purified by silica gel column (eluted with 20% EtOAc in petroleum ether) to afford the title compound 21-3 (490 mg, yield 73% over 2 steps).

[0170] Step 3: A mixture of compound 21-3 (480 mg, 1.19 mmol) in 4 N HCl (EtOAc solution, 5 mL) was stirred at room temperature for 2 h. After completion of the reaction, the mixture was concentrated to afford the title compound 21-4 (450 mg, crude) which was used at the next step without further purification.

[0171] Step 4: A solution of compound 21-4 (450 mg), compound 8-1 (297 mg, 1.43 mmol), DIPEA (390 mg, 3.02 mmol) and HATU (684 mg, 1.79 mmol) in DMF (5 mL) was stirred at room temperature for 16 h. After completion of the reaction, the mixture was diluted with EtOAc (50 mL) and washed with water (50 mL2) and brine (50 mL). The organic layer was dried by anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The crude product was purified by silica gel column (eluted with 30% EtOAc in petroleum ether) to afford the title compound 21-5 (339 mg, yield 58%). .sup.1H NMR (400 MHz, Chloroform-d) 7.17 (d, J=7.1 Hz, 2H), 7.11 (d, J=7.6 Hz, 2H), 5.30 (s, 1H), 4.14 (q, J=7.1 Hz, 4H), 3.52 (s, 4H), 3.49 (d, J=7.9 Hz, 1H), 2.99 (q, J=6.0 Hz, 2H), 2.57 (t, J=11.6 Hz, 1H), 2.45 (d, J=7.0 Hz, 2H), 1.83 (d, J=8.2 Hz, 3H), 1.76 (s, 1H), 1.60 (d, J=12.7 Hz, 2H), 1.50 (d, J=7.1 Hz, 3H), 1.25 (t, J=7.1 Hz, 8H), 1.11 (q, J=12.0 Hz, 2H), 0.91-0.87 (m, 6H). LCMS: m/z calculated for C.sub.28H.sub.44N.sub.2O.sub.5: 488.67; found: 489.75 [M+H].sup.+.

[0172] Step 5: To a solution of compound 21-5 (337 mg, 0.689 mmol) in EtOH (3 mL) and H.sub.2O (1.5 mL) was added NaOH (110 mg, 2.75 mmol) at 0 C. The reaction was stirred at room temperature for 1 h. The mixture was concentrated and adjusted to pH=2 with 1 N HCl at 0 C. The aqueous mixture was purified by reverse phase flash chromatography (C18 column, eluted with 55% acetonitrile in water, HCl condition). The desired components were lyophilized to afford the title compound 21-6 (207 mg, yield 64%) as a white solid.

[0173] Step 6: A mixture of compound 21-6 (155 mg, 0.33 mmol) and compound 9-7 (127 mg, 0.33 mmol) in toluene (3 mL) was stirred at 120 C. for 16 h. After completion of the reaction, the mixture was concentrated. The crude product was purified by pre-TLC (acetonitrile:dichloromethane=1:3) to afford 21 (115 mg, yield 44%) as a off-white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 9.08 (d, J=1.4 Hz, 1H), 8.89-8.85 (m, 2H), 8.73 (dd, J=2.5, 1.5 Hz, 1H), 7.92 (t, J=5.9 Hz, 1H), 7.46 (d, J=9.5 Hz, 1H), 7.32-7.28 (m, 2H), 7.25 (t, J=7.5 Hz, 2H), 7.21 (d, J=8.0 Hz, 2H), 7.19-7.15 (m, 1H), 7.05 (d, J=8.0 Hz, 2H), 4.72-4.64 (m, 1H), 4.31 (d, J=17.9 Hz, 1H), 3.84 (dd, J=17.0, 6.7 Hz, 2H), 3.62-3.52 (m, 2H), 3.25 (d, J=16.4 Hz, 2H), 3.07 (d, J=7.6 Hz, 2H), 2.86 (t, J=6.5 Hz, 2H), 2.39 (d, J=7.1 Hz, 2H), 1.77 (dt, J=13.7, 6.8 Hz, 1H), 1.74-1.62 (m, 3H), 1.61-1.36 (m, 6H), 1.31 (d, J=7.1 Hz, 4H), 1.16 (ddd, J=13.7, 10.4, 2.9 Hz, 1H), 1.09-0.97 (m, 1H), 0.86 (d, J=6.7 Hz, 3H), 0.83 (d, J=6.6 Hz, 6H), 0.78 (d, J=6.4 Hz, 3H). LCMS: m/z calculated for C.sub.43H.sub.57BN.sub.6O.sub.7: 780.77; found: 782.00 [M+H].sup.+.

Example 22

N-((S)-1-(((R)-3-methyl-1-(6-(4-((S)-2-(4-(2-(methyl-d.SUB.3.)propyl-3,3,3-d.SUB.3.)phenyl)propanamid o)butyl)-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)butyl)amino)-1-oxo-3-phenylpropan-2-yl)py razine-2-carboxamide (22)

##STR00074##

##STR00075##

[0174] Step 1: NaH (60% in mineral oil, 9.35 g, 233 mmol) was added portion-wise over 10 min to a solution of compound 22-2 (63 g, 281 mmol) in anhydrous THF (250 mL) at 0 C. under nitrogen atmosphere. After stirring at 0 C. for 1 h, to this was added deuterated acetone 22-1 (10 g, 156 mmol) slowly over 15 min. Then the reaction was warmed to room temperature and stirred for an additional 6 h. The mixture was quenched by saturated NH.sub.4Cl solution (300 mL) and extracted with isopropyl ether (300 mL2). The organic layers were dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to afford the title compound 22-3 (22 g, crude), which was used at the next step without further purification.

[0175] Step 2: 2 N LiOH (270 mL, 540 mmol) was added to a mixture of compound 22-3 (22 g, crude from Step 1) in EtOH (150 mL) at 0 C. The reaction was stirred at room temperature for 16 h. The resulting mixture was cooled to 0 C. and adjusted to pH=2 with 6 N aqueous HCl. The mixture was extracted with isopropyl ether (200 mL2). The organic layers were dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to afford the title compound 22-4 (13.4 g, 80% over 2 steps), which was used at the next step without further purification.

[0176] Step 3: To a solution of compound 22-4 (3 g, 28.2 mmol), compound 22-5 (6.92 g, 42.4 mmol) and DMAP (170 mg, 1.39 mmol) in DCM (50 mL) was added DCC (8.75 g, 42.4 mmol) at 0 C. The reaction was stirred at room temperature for 15 h. The mixture was concentrated and purified by silica gel column (eluted with 20% EtOAc in petroleum ether) to afford the title compound 22-6 (3.2 g, yield 45%) as a white solid.

[0177] Step 4: A mixture of compound 22-6 (2.3 g, 9.15 mmol), compound 22-7 (330 mg, 1.84 mmol) and B.sub.2(Pin).sub.2 (4.65 g, 18.3 mmol) in anhydrous PhCF.sub.3 (23 mL) was purged with nitrogen 3 times and stirred at 110 C. for 20 h. The mixture was concentrated and purified by silica gel column (eluted with 3% EtOAc in petroleum ether) to afford the title compound 22-8 (470 mg, yield 27%) as a colorless oil. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 4.98 (s, 1H), 1.19 (s, 12H).

[0178] Step 5: Under nitrogen, a mixture of compound 22-8 (550 mg, 2.92 mmol), compound 22-9 (736 mg, 3.21 mmol), Pd(PPh.sub.3).sub.4 (168 mg, 0.145 mmol) and K.sub.2CO.sub.3 (1.2 g, 8.68 mmol) in dioxane (6 mL) was stirred at 120 C. for 16 h. After completion of the reaction, the mixture was filtered and concentrated. The crude product was purified by reverse phase flash chromatography (C18 column, eluted with 50% acetonitrile in water, TFA condition). The desired components were lyophilized to afford the title compound 22-10 (460 mg, yield 75%) as a colorless slurry.

[0179] Step 6: A mixture of compound 22-10 (460 mg, 2.18 mmol) and PtO.sub.2 (50 mg, 0.22 mmol) in MeOH (5 mL) was stirred under hydrogen atmosphere for 16 h. After completion of the reaction, the mixture was filtered over celit and concentrated to afford the title compound 22-11 (350 mg, yield 75%) as a colorless slurry. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 7.18 (d, J=7.8 Hz, 2H), 7.08 (d, J=7.8 Hz, 2H), 3.60 (q, J=7.1 Hz, 1H), 2.40 (d, J=7.2 Hz, 2H), 1.76 (t, J=7.2 Hz, 1H), 1.32 (d, J=7.1 Hz, 3H).

[0180] Step 7: A solution of compound 13-2 (200 mg, 0.673 mmol), compound 22-11 (95 mg, 0.447 mmol), DIPEA (260 mg, 2.01 mmol) and HATU (300 mg, 0.789 mmol) in DMF (3 mL) was stirred at room temperature for 16 h. After completion of the reaction, the mixture was diluted with EtOAc (30 mL) and washed with water (30 mL2) and brine (30 mL). The organic layer was dried by anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The crude product was purified by silica gel column (eluted with 20% EtOAc in petroleum ether) to afford the title compound 22-12 (106 mg, yield 52%). LCMS: m/z calculated for C.sub.25H.sub.34D.sub.6N.sub.2O.sub.5: 454.64; found: 455.99 [M+H].sup.+.

[0181] Step 8: To a solution of compound 22-12 (100 mg, 0.220 mmol) in EtOH (2 mL) was added 2 N NaOH (0.44 mL, 0.88 mmol) at 0 C. and the reaction was stirred at room temperature for 1 h. The mixture was concentrated and adjusted to pH=2 with 1 N HCl at 0 C. The aqueous mixture was purified by reverse phase flash chromatography (C18 column, eluted with 35% acetonitrile in water, HCl condition). The desired components were lyophilized to afford the title compound 22-13 (68 mg, yield 75%) as a white solid.

[0182] Step 9: A mixture of compound 22-13 (60 mg, 0.151 mmol) and compound 9-7 (60 mg, 0.156 mmol) in toluene (3 mL) was stirred at 120 C. for 16 h. After completion of the reaction, the mixture was concentrated. The crude product was purified by pre-TLC (acetonitrile:dichloromethane=1:3) to afford 22 (26 mg, yield 23%) as an off-white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 9.12 (s, 1H), 8.90 (s, 2H), 8.76 (s, 1H), 8.00 (s, 1H), 7.38 (d, J=9.7 Hz, 1H), 7.27 (ddt, J=24.5, 17.1, 7.2 Hz, 7H), 7.09 (d, J=7.7 Hz, 2H), 4.72 (d, J=7.9 Hz, 1H), 4.10 (q, J=17.3 Hz, 2H), 3.95 (d, J=16.6 Hz, 1H), 3.58 (d, J=8.2 Hz, 2H), 3.50 (d, J=17.1 Hz, 1H), 3.17-3.01 (m, 6H), 2.41 (d, J=7.0 Hz, 2H), 1.78 (d, J=8.4 Hz, 1H), 1.66-1.54 (m, 2H), 1.43 (d, J=12.5 Hz, 4H), 1.35 (d, J=6.8 Hz, 3H), 1.20 (s, 1H), 0.87 (dd, J=16.0, 6.7 Hz, 6H). LCMS: m/z calculated for C.sub.40H.sub.47D.sub.6BN.sub.6O.sub.7: 746.75; found: 747.5 [M+H].sup.+.

Example 23

N-(4-((4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)oxy)butyl)-4-(4-iodophenyl)butana mide (23)

##STR00076##

##STR00077##

[0183] Step 1: To an ice-cold solution of compound 23-1 (10.0 g, 52.8 mmol) in anhydrous THF (200 mL) was added PPh.sub.3 (20.8 g, 79.3 mmol), followed by a solution of CBr.sub.4 (26.3 g, 79.3 mmol) in THF (100 mL). The reaction was allowed to warm to room temperature and stirred for 2 h. The mixture was filtered and concentrated. The residue was purified by silica gel column (eluted with 20% EtOAc in petroleum ether) to afford the title compound 23-2 (12 g, 90%) as a colorless oil.

[0184] Step 2: A mixture of compound 23-3 (500 mg, 1.95 mmol), K.sub.2CO.sub.3 (540 mg, 3.90 mmol) and compound 23-2 (590 mg, 2.34 mmol) in DMF (5 mL) was stirred at room temperature for 16 h. The resulting mixture was filtered and the cake washed with EtOAc (50 mL). The organic phase was washed with cold water (50 mL) and brine (50 mL). The organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The crude product was purified by silica gel column (eluted with 5% MeOH in dichloromethane) to afford the title compound 23-4 (585 mg, yield 70%) as a pale-yellow solid.

[0185] Step 3: 4N HCl (EtOAc solution, 2.5 mL) was added to a solution of compound 23-4 (585 mg, 1.36 mmol) in EtOAc (2 mL) and MeOH (0.5 mL). After the reaction was stirred at room temperature for 1 h, the mixture was concentrated to afford the title compound 23-5 (450 mg, yield 90%) as a white solid.

[0186] Step 4: A mixture of compound 23-5 (80 mg, 0.275 mmol), DIPEA (56 mg, 0.434 mmol), DMAP (3 mg, 0.024 mmol), EDCI (65 mg, 0.340 mmol) and compound 1-2 (80 mg, 0.220 mmol) in DCM (5 mL) was stirred at room temperature for 1 h. After completion of the reaction, the mixture was concentrated. The crude product was purified by pre-HPLC (eluted with 60% acetonitrile in water, HCl condition) to afford 23 (60 mg, yield 45%) as a white solid. .sup.1H NMR (500 MHz, DMSO-d.sub.6) 8.19 (d, J=8.1 Hz, 1H), 7.90 (d, J=5.4 Hz, 1H), 7.84 (d, J=8.4 Hz, 1H), 7.77-7.72 (m, 1H), 7.64-7.56 (m, 3H), 7.00 (d, J=8.1 Hz, 2H), 4.41 (t, J=6.6 Hz, 2H), 3.15 (q, J=6.5 Hz, 2H), 3.00 (t, J=7.6 Hz, 2H), 2.52 (d, J=7.9 Hz, 2H), 2.07 (t, J=7.4 Hz, 2H), 1.92 (t, J=7.5 Hz, 2H), 1.82 (t, J=7.6 Hz, 2H), 1.77 (t, J=7.5 Hz, 2H), 1.71-1.62 (m, 2H), 1.42 (q, J=7.4 Hz, 2H), 0.93 (t, J=7.4 Hz, 3H). LCMS: m/z calculated for C.sub.28H.sub.34IN.sub.5O.sub.2: 599.52; found: 600.82 [M+H].sup.+.

Example 24

N-(4-((4-amino-2-butyl-7-(dimethylphosphoryl)-1H-imidazo[4,5-c]quinolin-1-yl)oxy)butyl)-4-(4-iodophenyl)butanamide (24)

##STR00078##

##STR00079##

[0187] Step 1: A mixture of compound 24-1 (19 mg, 0.048 mmol), compound 1-2 (28 mg, 0.0965 mmol), EDCI (36 mg, 0.188 mmol), DMAP (1 mg, 0.008 mmol) and TEA (10 mg, 0.099) in DCM (0.5 ml) and DMF (0.5 ml) was stirred at room temperature for 1 h. After completion of the reaction, the resulting mixture was filtered and purified by pre-HPLC (eluted with 70% acetonitrile in water, HCl condition). The components were lyophilized to give 24 (11.2 mg, 35.8% yield) as a yellow oil. LCMS: m/z calculated for C.sub.30H.sub.39IN.sub.5O.sub.3P: 675.55; found: 676.2 [M+H].sup.+.

Example 25

N-(2-((2-((4-((4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)oxy)butyl) (3-(dimethylamino)propyl)amino)-3,4-dioxocyclobut-1-en-1-yl)amino)ethyl)-4-(4-iodopheny l)butanamide (25)

##STR00080##

##STR00081##

[0188] Step 1: To a solution of compound 1-2 (400 mg, 1.37 mmol) in DCM (8 mL) was added EDCI (395 mg, 2.06 mmol) and compound 9-1 (243 mg, 1.51 mmol). After stirring at RT for 2 h, TLC showed compound 1-2 was consumed. The reaction mixture was diluted with EtOAc and washed with 1 N HCl, sat. Na.sub.2CO.sub.3 solution and brine. The organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure to afford the title compound 25-1 (330 mg) as a colorless oil, which was used directly at the next step. TLC (Petroleum ether:EtOAc/1:1): R.sub.f (compound 9-1)=0.55; R.sub.f (compound 25-1)=0.37.

[0189] Step 2: A mixture of crude compound 25-1 (330 mg) in HCl/EtOAc (4 M, 10 mL) was stirred at RT for 30 minutes. After completion of the reaction, the mixture was concentrated. The residue was re-dissolved in EtOAc and washed with saturated Na.sub.2CO.sub.3 solution. The organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure to afford the title compound 25-2 (225 mg) as a colorless oil, which was used directly at the next step. TLC (Petroleum ether:EtOAc/1:1) R.sub.f (compound 25-1)=0.37; TLC (DCM:MeOH/10:1): R.sub.f (compound 25-2)=0.46.

[0190] Step 3: To a solution of compound 25-3 (154 mg, 0.905 mmol) in MeOH (5 mL) was added dropwise a solution of compound 25-2 (225 mg, crude) in MeOH (2 mL) at 0 C. After stirring at RT for 4 h, TLC showed compound 25-2 was consumed and a new spot with lower polarity was formed. The reaction mixture was evaporated in vacuum and purified by silica gel column (eluted with 5% MeOH in DCM) to afford the title compound 25-4 (230 mg, yield 36% over 3 steps) as a white solid. TLC (DCM:MeOH/10:1): R.sub.f (compound 25-2)=0.46, R.sub.f(compound 25-4)=0.58.

[0191] Step 4: A mixture of compound 25-4 (67 mg, 0.145 mmol), compound 25-5 (40 mg, 0.970 mmol) and DIPEA (40 mg, 0.396 mmol) in acetonitrile (2 mL) and DCM (2 mL) was stirred at RT for 24 h. LC-MS showed compound 25-5 was consumed and one new peak with desired m/z was detected by LCMS. The reaction mixture was concentrated, re-dissolved in acetonitrile and water, and purified by HPLC (C18 column, eluted with acetonitrile/H.sub.2O, HCl condition). The desired component was lyophilized to give 25 (8 mg, yield 10%) as a white powder. .sup.1H NMR (400 MHz, Methanol-d.sub.4) 8.12 (d, J=8.1 Hz, 1H), 7.68 (d, J=8.4 Hz, 1H), 7.53 (t, J=7.8 Hz, 1H), 7.46 (d, J=7.9 Hz, 2H), 7.36 (t, J=7.7 Hz, 1H), 6.86 (d, J=7.9 Hz, 2H), 4.33 (s, 2H), 3.75 (t, J=5.8 Hz, 2H), 3.41 (s, 2H), 3.35 (d, J=2.4 Hz, 2H), 2.99 (t, J=7.6 Hz, 2H), 2.47 (t, J=7.8 Hz, 2H), 2.39 (d, J=7.1 Hz, 2H), 2.27 (s, 6H), 2.16 (d, J=7.7 Hz, 2H), 1.98 (d, J=9.8 Hz, 2H), 1.85 (dq, J=23.9, 7.5 Hz, 8H), 1.49 (d, J=7.6 Hz, 2H), 1.30-1.26 (m, 2H), 1.01 (t, J=7.4 Hz, 3H). LCMS: m/z calculated for C.sub.39H.sub.51IN.sub.8O.sub.4: 822.79; found: 823.4 [M+H].sup.+.

Example 26

(S)-N-(2-((2-((4-((4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)oxy)butyl)amino)-3,4-dio xocyclobut-1-en-1-yl)amino)ethyl)-2-(4-isobutylphenyl)propanamide (26)

##STR00082##

##STR00083##

[0192] Step 1: To a solution of compound 25-3 (1.06 g, 6.23 mmol) in anhydrous MeOH (10 mL) was added dropwise a solution of compound 9-1 (1 g, 6.24 mmol) in anhydrous MeOH (10 mL). The mixture was stirred at room temperature for 1 h. After completion of the reaction, the mixture was diluted with EtOAc (50 mL), washed with water (50 mL) and brine (50 mL). The organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The crude product was purified by column (eluted with 40% EtOAc in petroleum ether) to afford the title compound 26-1 (1.63 g, yield 97%) as a colorless oil.

[0193] Step 2: To a solution of compound 23-5 (339 mg, 1.03 mmol) and DIPEA (333 mg 2.58 mmol) in anhydrous MeOH (4 mL) was added dropwise a solution of compound 26-1 (440 mg, 1.55 mmol) in anhydrous MeOH (3 mL). The mixture was stirred at room temperature for 3 h. After completion of the reaction, the mixture was concentrated. The crude product was purified by reverse phase flash chromatography (C18 column, eluted with 60% acetonitrile in water, TFA condition). The desired components were lyophilized to give the title compound 26-2 (TFA salt, 445 mg, yield 63%) as a yellow solid.

[0194] Step 3: A mixture of compound 26-2 (445 mg, 0.65 mmol) in a solution of 20% TFA in DCM (3 mL) was stirred at room temperature for 1 h. After completion of the reaction, the mixture was concentrated to get rid of organic solvents to afford the title compound 26-3 (420 mg, yield 92%) as a pale-yellow slurry.

[0195] Step 4: A mixture of compound 26-3 (100 mg, 0.148 mmol), compound 8-1 (46 mg, 0.222 mmol), DMAP (2 mg, 0.016 mmol), DIPEA (57 mg, 0.442 mmol) and EDCI (42 mg, 0.220 mmol) in DMF (2 mL) was stirred at room temperature for 16 h. After completion of the reaction, the mixture was filtered and purified by pre-HPLC (eluted with 50% acetonitrile in water, HCl condition). The desired components were lyophilized to afford 26 (40 mg, 39% yield) as a white solid. .sup.1H NMR (600 MHz, DMSO-d.sub.6) 8.20 (dd, J=8.1, 1.4 Hz, 1H), 7.85-7.81 (m, 1H), 7.74 (ddd, J=8.4, 7.2, 1.3 Hz, 1H), 7.62 (t, J=7.7 Hz, 1H), 7.17 (d, J=7.5 Hz, 2H), 7.01 (d, J=7.2 Hz, 2H), 4.44 (t, J=6.5 Hz, 2H), 3.57-3.43 (m, 5H), 3.28 (d, J=7.0 Hz, 1H), 3.12 (dd, J=13.3, 6.5 Hz, 1H), 3.00 (t, J=7.6 Hz, 2H), 2.39-2.31 (m, 2H), 2.01 (s, 2H), 1.82 (td, J=14.1, 13.2, 6.6 Hz, 4H), 1.76 (d, J=4.4 Hz, 1H), 1.43 (q, J=7.4 Hz, 2H), 1.29 (d, J=7.0 Hz, 3H), 0.93 (t, J=7.4 Hz, 3H), 0.81 (d, J=6.6 Hz, 6H). LCMS: m/z calculated for C.sub.37H.sub.47N.sub.7O.sub.4: 653.83; found: 654.88 [M+H].sup.+.

Example 27

(S)-N-(4-((4-amino-2-butyl-7-(dimethylphosphoryl)-1H-imidazo[4,5-c]quinolin-1-yl)oxy)but yl)-2-(4-isobutylphenyl)propanamide (27)

##STR00084##

##STR00085##

[0196] Step 1: A mixture of compound 24-1 (60 mg, 0.148 mmol), compound 8-1 (46 mg, 0.223 mmol), DMAP (2 mg, 0.016 mmol), DIPEA (57 mg, 0.442 mmol) and EDCI (45 mg, 0.235 mmol) in DMF (2 mL) was stirred at room temperature for 16 h. After completion of the reaction, the mixture was filtered and purified by pre-HPLC (eluted with 45% acetonitrile in water, HCl condition). The desired components were lyophilized to afford 27 (24 mg, 27% yield) as a white solid. .sup.1H NMR (600 MHz, DMSO-d.sub.6) 8.29-8.24 (m, 2H), 8.09 (t, J=5.7 Hz, 1H), 7.95 (ddd, J=10.0, 8.3, 1.3 Hz, 1H), 7.23 (d, J=8.1 Hz, 2H), 7.05 (d, J=8.1 Hz, 2H), 4.39 (t, J=6.6 Hz, 2H), 3.60 (s, 1H), 3.16 (q, J=6.6 Hz, 2H), 3.04-3.00 (m, 2H), 2.37 (d, J=7.1 Hz, 2H), 1.91-1.82 (m, 4H), 1.77 (d, J=13.5 Hz, 7H), 1.68-1.63 (m, 2H), 1.49-1.41 (m, 2H), 1.32 (d, J=7.1 Hz, 3H), 0.96 (t, J=7.4 Hz, 3H), 0.82 (s, 6H). LCMS: m/z calculated for C.sub.33H.sub.46N.sub.5O.sub.3P: 591.74; found: 592.93 [M+H].sup.+.

Example 28

(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)methyl 4-(4-iodophenyl) butanoate (28)

##STR00086##

##STR00087##

[0197] Step 1: Compound 28-1 (1.3 g, 10.0 mmol) was dissolved in 37% formaldehyde solution (1.8 g, 22.2 mmol) and the mixture was refluxed with stirring at 60 C. for 4 h. The mixture was concentrated to give the title compound 28-2 (1.5 g, yield 93%) as a colorless oil, which was used directly at the next step without purification.

[0198] Step 2: Oxalyl chloride (0.50 mL, 5.82 mmol) and catalytic amount DMF were added to a solution of compound 1-2 (200 mg, 0.69 mmol) in dichloromethane (20 mL). The resulting mixture was stirred at 45 C. for 1 h. The mixture was azeotroped with dichloromethane3 times under reduced pressure to give the crude chloride intermediate. The chloride was re-dissolved in anhydrous dichloromethane (5 mL) and added to a solution of compound 28-2 (220 mg, 1.38 mmol), DMAP (8 mg, 0.07 mmol) and TEA (100 uL, 0.69 mmol) in anhydrous dichloromethane (10 mL) at room temperature. The resulting mixture was refluxed at 50 C. with stirring for 10 h. After the mixture was concentrated, the residue was purified by silica gel column (eluted with 50% EtOAc in petroleum ether) to afford 28 (72 mg, 26% yield) as a white solid. .sup.1H NMR (400 MHz, DMSO-d6) 12.00 (s, 1H), 8.14 (d, J=6.4 Hz, 1H), 7.68-7.56 (m, 2H), 7.01 (d, J=7.8 Hz, 2H), 5.56 (s, 2H), 2.54 (d, J=7.6 Hz, 2H), 2.33 (t, J=7.3 Hz, 2H), 1.78 (t, J=7.5 Hz, 2H).

Example 29

(5-fluoro-2,4-dioxopyrimidine-1,3(2H,4H)-diyl)bis(methylene)bis(4-(4-iodo-1-phenyl)butanoate) (29)

##STR00088##

##STR00089##

[0199] Step 1: A suspension of compound 28-1 (1.0 g, 7.69 mmol) in 37% formaldehyde solution (2 mL) was stirred at 60 C. for 4 h. The reaction mixture was concentrated to afford the title compound 29-1 (1.5 g) as a colorless oil, which was used directly at the next step without purification.

[0200] Step 2: To a solution of compound 1-2 (500 mg, 1.73 mmol) in DCM (5 mL) was added oxalyl chloride (2.18 g, 17.21 mmol) and catalytic amount DMF at room temperature. The resulting mixture was stirred at 45 C. for 1 h. After compound 1-2 was consumed, the mixture was concentrated. The residue was re-dissolved in DCM (10 mL), to this solution was added compound 29-1 (250 mg, 1.32 mmol), DMAP (16 mg, 0.131 mmol) and TEA (650 mg, 6.44 mmol) at room temperature. The resulting mixture was stirred at 50 C. for 16 h. After completion of the reaction, the mixture was concentrated. The residue was purified by silica gel column (eluted with 2% EtOAc in petroleum ether) and further purified by reverse phase flash chromatography (C18 column, eluted with acetonitrile and water, TFA condition) to afford 29 (70 mg, 7% yield over 2 steps) as a white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 8.29 (dq, J=6.3, 3.8, 3.1 Hz, 1H), 7.61 (ddq, J=6.4, 4.1, 2.4 Hz, 4H), 6.99 (dt, J=8.1, 3.4 Hz, 4H), 5.82-5.75 (m, 2H), 5.67-5.58 (m, 2H), 2.52 (d, J=5.9 Hz, 4H), 2.35-2.23 (m, 4H), 1.76 (dq, J=12.2, 7.2 Hz, 4H). LCMS: m/z calculated for C.sub.26H.sub.25FI.sub.2N.sub.2O.sub.6: 734.30; found: 757.33. [M+Na].sup.+.

Example 30

(2aR,4S,4aS,6R,9S,11 S,12S,12aR,12bS)-12b-acetoxy-9-(((2R,3S)-3-((tert-butoxycarbonyl)amino)-2-((4-(4-iodophenyl)butanoyl)oxy)-3-phenylpropanoyl)oxy)-4,6,11-trihydroxy-4a,8,13,13-tetramethyl-5-oxo-2a,3,4,4a,5,6,9,10,11,12,12a,12b-dodecahydro-1H-7,11-methanocyclodeca[3,4]benzo[1,2-b]oxet-12-yl benzoate (30)

##STR00090##

##STR00091##

[0201] Step 1: DCC (77 mg, 0.37 mmol) and DMAP (45 mg, 0.37 mmol) were added to a solution of compound 30-1 (200 mg, 0.25 mmol) and 4-(p-iodophenyl)butyric acid 1-2 (79 mg, 0.27 mmol) in dichloromethane (16 mL) at 10 C. After stirring at 10 C. for 3 h, the mixture was filtered, diluted with EtOAc (50 mL), and washed with water (50 mL) and brine (50 mL). The organic layer was dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The residue was purified by silica gel column (eluted with 40% EtOAc in petroleum ether) to afford 30 (115 mg, yield: 43%) as a white solid. .sup.1H NMR (400 MHz, Chloroform-d) 8.16 (d, J=7.7 Hz, 2H), 7.68-7.60 (m, 3H), 7.55 (t, J=7.5 Hz, 2H), 7.43 (t, J=7.5 Hz, 2H), 7.37-7.31 (m, 3H), 6.90 (d, J=7.8 Hz, 2H), 6.30 (s, 1H), 5.74 (d, J=7.0 Hz, 1H), 5.52 (s, 1H), 5.45-5.34 (m, 2H), 5.26 (s, 1H), 5.01 (d, J=9.4 Hz, 1H), 4.37 (d, J=8.5 Hz, 1H), 4.31 (q, J=8.3 Hz, 1H), 4.24 (d, J=6.4 Hz, 2H), 4.20-4.13 (m, 1H), 3.99 (d, J=7.0 Hz, 1H), 2.64 (dt, J=15.4, 8.1 Hz, 1H), 2.54 (t, J=7.6 Hz, 3H), 2.49 (s, 3H), 2.38 (dp, J=23.3, 8.3 Hz, 4H), 2.22 (s, 1H), 2.09 (d, J=1.7 Hz, 2H), 2.01 (s, 3H), 1.90 (q, J=8.4, 7.7 Hz, 4H), 1.80 (s, 3H), 1.69 (s, 1H), 1.63 (d, J=1.7 Hz, 3H), 1.54 (d, J=7.5 Hz, 1H), 1.38 (s, 11H), 1.32 (d, J=1.9 Hz, 1H), 1.30 (d, J=1.7 Hz, 2H), 1.28 (s, 4H), 1.17 (s, 4H). LCMS: m/z calculated for C.sub.53H.sub.62INO.sub.15: 1079.98; found: 1102.7 [M+Na].sup.+.

Example 31

N.SUP.2.-(4-(2-(2-amino-4-oxo-4,7-dihydro-31H-pyrrolo[2,3-d]pyrimidin-5-yl)ethyl) benzoyl)-N.SUP.5.-(2-(4-(4-iodophenyl)butanamido)ethyl)-L-glutamine (31)

##STR00092##

##STR00093##

[0202] Step 1: EDCI (362 mg, 1.89 mmol), triethylamine (0.74 mL, 5.16 mmol) and DMAP (20 mg, 0.17 mmol) were added to a solution of compound 31-1 (520 mg, 1.41 mmol) and compound 25-2 (574 mg, 1.89 mmol) in dichloromethane (20 mL) at 0 C. The reaction was stirred at room temperature for 2 h, the mixture was diluted with dichloromethane (100 mL), washed with 1N HCl (100 mL) and brine (100 mL). The organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to afford the title compound 31-2 (740 mg, yield: 85%), which was used at the next step without further purification.

[0203] Step 2: A mixture of compound 31-2 (400 mg, 0.65 mmol) in 4 N HCl (EtOAc solution, 5 mL) was stirred at room temperature for 1 h. A white precipitation was formed and filtered. The solid was dried in vacuum to afford the title compound 31-3 (HCl salt, 200 mg, yield 62%).

[0204] Step 3: A solution of compound 31-4 (200 mg, 0.67 mmol), DMAP (8 mg, 0.07 mmol), NHS (115 mg, 1.00 mmol) and EDCI (128 mg, 0.67 mmol) in anhydrous DMSO (3 mL) was stirred at room temperature for 1 h. Compound 31-3 (200 mg, 0.40 mmol) and Et.sub.3N (483 uL, 3.35 mmol) were added. After stirring at room temperature for 12 h, the mixture was purified by prep-HPLC (eluted with 60% acetonitrile in water, HCl condition) to give 31 (67 mg, yield: 22%) as a light pink solid. .sup.1H NMR (400 MHz, DMSO-d6) 10.71 (s, 1H), 8.60 (d, J=7.7 Hz, 1H), 7.89 (s, 1H), 7.79 (d, J=7.7 Hz, 3H), 7.61 (d, J=7.9 Hz, 2H), 7.29 (d, J=7.9 Hz, 2H), 7.00 (d, J=8.0 Hz, 2H), 6.34 (s, 1H), 6.27 (s, 1H), 4.32 (d, J=10.2 Hz, 1H), 3.12-2.80 (m, 12H), 2.11 (dt, J=69.0, 7.7 Hz, 7H), 1.79-1.67 (m, 2H). LCMS: m/z calculated for C.sub.32H.sub.36IN.sub.7O.sub.6: 741.59; found: 742.4. [M+H].sup.+.

Example 32

(S)-2-(4-(2-(2-amino-4-oxo-4,7-dihydro-3H-pyrrolo[2,3-d]pyrimidin-5-yl)ethyl) benzamido)-5-(2-(4-(4-iodophenyl)butanamido)ethoxy)-5-oxopentanoic acid (32)

##STR00094##

##STR00095##

[0205] Step 1: To a mixture of acid 1-2 (500 mg, 1.72 mmol) and NHS (295 mg, 2.56 mmol) in DCM (10 mL) were added EDCI (658 mg, 3.44 mmol) and DMAP (21 mg, 0.172 mmol), the mixture was stirred at room temperature for 2 h. The reaction mixture was diluted with DCM (50 mL) and washed with 1 N HCl (50 mL). The organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to give the crude activated NHS ester as a white solid. To a solution of compound 32-1 (126 mg, 2.06 mmol) and DIEA (340 uL, 2.06 mmol) in DMF (5 mL) was added dropwise a solution of activated NHS ester in THF (5 mL). The reaction mixture was allowed to stir at room temperature for 16 h. The resulting reaction mixture was diluted with EtOAc (50 mL) and washed with 1 N HCl (50 mL) and brine. The organic layers were dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to give the title compound 32-2 (495 mg, 86% yield) as a white solid, which was used at the next step without further purification.

[0206] Step 2: EDCI (218 mg, 1.14 mmol) was added to a solution of DMAP (7 mg, 0.06 mmol), compound 32-2 (190 mg, 0.57 mmol) and compound 31-1 (260 mg, 0.86 mmol) in DCM (10 mL) at 0 C. After stirring at room temperature for 2 h, the mixture was diluted with DCM (100 mL), washed with 1N HCl (50 mL) and brine (50 mL). The organic layer was dried over Na.sub.2SO.sub.4, filtered and concentrated to afford the title compound 32-3 (845 mg, 85% yield), which was used at the next step without further purification.

[0207] Step 3: Compound 32-3 (400 mg, 0.65 mmol) was dissolved in DCM (3 mL) and TFA (3 mL), the reaction mixture was stirred at room temperature for 2 h. The mixture was concentrated to afford the title compound 32-4 (240 mg, 91% yield) as a pale-yellow oil.

[0208] Step 4: A mixture of compound 31-4 (200 mg, 0.67 mmol), DMAP (8 mg, 0.07 mmol) and NHS (115 mg, 1.00 mmol) and EDCI (128 mg, 0.67 mmol) in anhydrous DMSO (3 mL) was stirred at room temperature for 1 h. To this mixture was added compound 32-4 (120 mg, 0.26 mmol) and Et.sub.3N (0.483 mL, 3.35 mmol). The mixture was stirred at room temperature for 12 h and purified by prep-HPLC (eluted with 60% acetonitrile in water, HCl condition) to afford 32 (55 mg, 28% yield) as a white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 8.60 (d, J=7.8 Hz, 1H), 8.01-7.90 (m, 1H), 7.79 (d, J=7.8 Hz, 2H), 7.67-7.58 (m, 2H), 7.31 (d, J=7.8 Hz, 2H), 7.01 (d, J=7.8 Hz, 2H), 6.34 (s, 1H), 4.45 (dd, J=9.3, 5.5 Hz, 1H), 4.03 (q, J=5.7 Hz, 2H), 3.28 (t, J=5.6 Hz, 2H), 3.00 (t, J=7.7 Hz, 2H), 2.88 (t, J=7.5 Hz, 2H), 2.46 (dd, J=19.7, 7.7 Hz, 4H), 2.22-2.11 (m, 1H), 2.07 (t, J=7.4 Hz, 2H), 2.04-1.94 (m, 1H), 1.77 (q, J=7.4 Hz, 2H). LCMS: m/z calculated for C.sub.32H.sub.35IN.sub.6O.sub.7: 742.57; found: 743.3. [M+H].sup.+.

Example 33

(S)-2-(4-(2-(2-amino-4-oxo-4,7-dihydro-3H-pyrrolo[2,3-d]pyrimidin-5-yl)ethyl) benzamido)-5-(4-((S)-2-(4-isobutylphenyl)propanoyl)piperazin-1-yl)-5-oxopentanoic acid (33)

##STR00096##

##STR00097##

[0209] Step 1: EDCI (3.7 g, 19.37 mmol) was added to a mixture of acid 8-1 (2.00 g, 9.69 mmol), amine 33-1 (2.70 g, 14.49 mmol) and DMAP (118 mg, 0.967 mmol) in DCM (20 mL), the mixture was stirred at room temperature for 4 h. The reaction mixture was diluted with DCM (50 mL), washed with 1 N HCl (50 mL2) and brine (50 mL2). The organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to give the title compound 33-2 (3.62 g, crude) as a white solid, which was used at the next step without further purification. LCMS: m/z calculated for C.sub.22H.sub.34N.sub.2O.sub.3: 374.53; found: 397.47. [M+Na].sup.+.

[0210] Step 2: To a solution of compound 33-2 (3.62 g, crude) in EtOAc (30 mL) was added 4 N HCl in EtOAc (30 mL). The mixture was stirred at room temperature for 1 h. The mixture was diluted with EtOAc (30 mL) and filtered. The precipitation was washed with EtOAc and dried to afford the title compound 33-3 (2.7 g, yield 89% over 2 steps) as a white solid. LCMS: m/z calculated for C.sub.17H.sub.26N.sub.2O: 274.41; found: 275.90. [M+H].sup.+.

[0211] Step 3: EDCI (3.12 g, 16.33 mmol) was added to a solution of DMAP (200 mg, 1.64 mmol), TEA (1.75 g, 17.32 mmol), compound 33-3 (2.70 g, 8.68 mmol) and compound 31-1 (2.48 g, 8.17 mmol) in DCM (40 mL) at 0 C. After stirring at room temperature for 2 h, the mixture was diluted with DCM (100 mL), washed with 1N HCl (50 mL) and brine (50 mL). The organic layer was dried over Na.sub.2SO.sub.4, filtered and concentrated. The crude product was purified by silica gel column (eluted with 50% EtOAc in petroleum ether) to afford the title compound 33-4 (3.66 g, 80% yield) as a white solid. LCMS: m/z calculated for C.sub.31H.sub.49N.sub.3O.sub.6: 559.75; found: 582.98. [M+Na].sup.+.

[0212] Step 4: Compound 33-4 (2.88 g, 5.00 mmol) was dissolved in DCM (10 mL) and TFA (10 mL), the reaction was stirred at room temperature for 2 h. The mixture was concentrated to afford the title compound 33-5 (2.6 g, 90% yield) as a pale-yellow oil. LCMS: m/z calculated for C.sub.22H.sub.33N.sub.3O.sub.4: 403.52; found: 404.95. [M+H].sup.+.

[0213] Step 5: A mixture of compound 31-4 (300 mg, 1.00 mmol), DMAP (12 mg, 0.098 mmol) and NHS (174 mg, 1.51 mmol) and EDCI (383 mg, 2.00 mmol) in anhydrous DMSO (3 mL) was stirred at room temperature for 2 h. After the activated NHS ester was formed completely, compound 33-5 (560 mg, 1.08 mmol) and Et.sub.3N (508 mg, 5.02 mmol) was added. The mixture was stirred at 30 C. for 12 h and purified by prep-HPLC (eluted with 60% acetonitrile in water, TFA condition) to afford 33 (43 mg, 4% yield) as a white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6 and D.sub.2O) 7.75 (dd, J=8.0, 3.9 Hz, 2H), 7.28 (d, J=8.1 Hz, 2H), 7.13 (d, J=6.6 Hz, 2H), 7.08 (d, J=7.6 Hz, 2H), 6.37 (s, 1H), 4.34 (s, 1H), 4.05 (q, J=6.7 Hz, 1H), 3.58-3.48 (m, 4H), 3.48-3.34 (m, 4H), 2.97 (dd, J=9.3, 6.2 Hz, 2H), 2.86 (dd, J=9.6, 5.9 Hz, 2H), 2.38 (d, J=6.5 Hz, 4H), 1.99 (dt, J=40.0, 6.8 Hz, 2H), 1.84-1.71 (m, 1H), 1.26 (d, J=6.6 Hz, 3H), 0.81 (t, J=6.1 Hz, 6H). LCMS: m/z calculated for C.sub.37H.sub.45N.sub.7O.sub.6: 683.81; found: 684.65. [M+H].sup.+.

Example 34

N2-(4-(2-(2-amino-4-oxo-4,7-dihydro-3H-pyrrolo[2,3-d]pyrimidin-5-yl)ethyl) benzoyl)-N.SUB.5.-(2-((S)-2-(4-isobutylphenyl)propanamido)ethyl)-L-glutamine (34)

##STR00098##

##STR00099##

[0214] Step 1: EDCI (3.71 g, 19.42 mmol) was added to a mixture of acid 8-1 (2.00 g, 9.69 mmol), amine 9-1 (2.33 g, 14.54 mmol) and DMAP (238 mg, 1.95 mmol) in DCM (20 mL), the mixture was stirred at room temperature for 4 h. The reaction mixture was diluted with DCM (50 mL), washed with 1 N HCl (50 mL2) and brine (50 mL2). The organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to give the title compound 34-1 (3.2 g, crude) as a white solid, which was used at the next step without further purification. LCMS: m/z calculated for C.sub.20H.sub.32N.sub.2O.sub.3: 348.49; found: 397.47. [M+Na].sup.+.

[0215] Step 2: To a solution of compound 34-1 (3.2 g, crude) in EtOAc (30 mL) was added 4 N HCl in EtOAc (30 mL), the mixture was stirred at room temperature for 1 h to form a white precipitation completely. The mixture was diluted with EtOAc (30 mL) and filtered. The precipitation was washed with EtOAc and dried to afford the title compound 34-2 (1.9 g, yield 68% over 2 steps) as a white solid. LCMS: m/z calculated for C.sub.15H.sub.24N.sub.2O: 248.37; found: 249.90. [M+H].sup.+.

[0216] Step 3: EDCI (1.28 g, 6.70 mmol) was added to a solution of DMAP (82 mg, 0.672 mmol), TEA (680 mg, 6.73 mmol), compound 34-2 (1.0 g, 3.51 mmol) and compound 31-1 (1.01 g, 3.32 mmol) in DCM (20 mL) at 0 C. The reaction was stirred at room temperature for 2 h, the mixture was diluted with DCM (50 mL), washed with 1N HCl (50 mL) and brine (50 mL). The organic layer was dried over Na.sub.2SO.sub.4, filtered and concentrated. The crude product was purified by silica gel column (eluted with 50% EtOAc in petroleum ether) to afford the title compound 34-3 (953 mg, 53% yield) as a white solid. LCMS: m/z calculated for C.sub.29H.sub.47N.sub.3O.sub.6: 533.71; found: 556.98. [M+Na].sup.+.

[0217] Step 4: Compound 34-3 (700 mg, 1.31 mmol) was dissolved in DCM (7 mL) and TFA (7 mL), the reaction was stirred at room temperature for 2 h. The mixture was concentrated to afford the title compound 34-4 (700 mg, 90% yield) as a pale-yellow oil. LCMS: m/z calculated for C.sub.20H.sub.31N.sub.3O.sub.4: 377.49; found: 378.88. [M+H].sup.+.

[0218] Step 5: A mixture of compound 31-4 (135 mg, 0.452 mmol), DMAP (5 mg, 0.0409 mmol) and NHS (78 mg, 0.678 mmol) and EDCI (172 mg, 0.900 mmol) in anhydrous DMSO (3 mL) was stirred at room temperature for 2 h. After the NHS activated ester was formed completely, compound 34-4 (310 mg, 0.630 mmol) and Et.sub.3N (230 mg, 2.27 mmol) was added. The mixture was stirred at 30 C. for 12 h and purified by prep-HPLC (eluted with 60% acetonitrile in water, TFA condition) to afford 34 (26 mg, 6% yield) as a white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6 and D.sub.2O) 7.81 (dd, J=18.1, 7.9 Hz, 2H), 7.30 (dd, J=11.5, 7.9 Hz, 2H), 7.19-7.15 (m, 2H), 7.06-7.02 (m, 2H), 6.36 (s, 1H), 4.33 (dd, J=10.2, 4.6 Hz, 1H), 3.14-3.00 (m, 4H), 2.99-2.94 (m, 2H), 2.86 (dd, J=9.3, 5.8 Hz, 2H), 2.37 (d, J=7.1 Hz, 2H), 2.18 (t, J=7.5 Hz, 2H), 2.14-1.86 (m, 2H), 1.82-1.70 (m, 1H), 1.28 (d, J=6.9 Hz, 3H), 0.84-0.79 (m, 6H). LCMS: m/z calculated for C.sub.35H.sub.43N.sub.7O.sub.6: 657.77; found: 658.86. [M+H].sup.+.

Example 35

trans-[Pt(DACH)(ox)(OH)(IPBA)](35, IPBA=4-(4-iodophenyl)butanoic acid) (35)

##STR00100##

##STR00101##

[0219] Step 1: Hydrogen peroxide (30%, 15 mL) was added dropwise to a suspension of oxaliplatin 35-1 (200 mg, 0.50 mmol) in water (5 ml). After addition, the reaction mixture was heated to 75 C. and stirred for 5 hours. A clear solution was formed and cooled to room temperature. The resulting solution was concentrated. The residue was washed with EtOH and MTBE to give the title compound 35-2 (170 mg, yield 78%) as a yellow solid.

[0220] Step 2: To a solution of acid 1-2 (114 mg, 0.39 mmol) and TBTU (127 mg, 0.39 mmol) in anhydrous DMSO (5 mL) was added TEA (55 uL, 0.39 mmol). The mixture was intensively stirred at room temperature for 15 min. Compound 35-2 (170 mg, 0.39 mmol) was added and the reaction mixture was stirred at 60 C. for 16 h. The resulting reaction mixture was filtered to remove un-reacted solid. The clear solution was purified by reverse phase flash chromatography (C18 column, eluted with 50% acetonitrile in water, neutral condition). The desired components were lyophilized overnight to afford 35 (50.8 mg, yield 18%) as a white solid. .sup.1H NMR (400 MHz, DMSO-d6) 7.59 (d, J=7.8 Hz, 2H), 6.97 (d, J=7.8 Hz, 2H), 2.44 (d, J=7.6 Hz, 2H), 2.15 (d, J=7.5 Hz, 2H), 2.06 (t, J=17.4 Hz, 2H), 1.68 (t, J=7.5 Hz, 2H), 1.56-0.99 (m, 8H).

Example 36

trans-[Pt(DACH)(ox)(IPBA).SUB.2.](36, IPBA=4-(4-iodophenyl)butanoic acid) (36)

##STR00102##

##STR00103##

[0221] Step 1: A mixture of compound 1-2 (587 mg, 2.03 mmol), TBTU (650 mg, 2.02 mmol) and TEA (205 mg, 2.02 mmol) in DMF (5 mL) was stirred at room temperature under nitrogen for 15 min. To this mixture was added compound 35-2 (218 mg, 0.506 mmol) in one portion. The resulting reaction was stirred at 60 C. for 16 h, the mixture was filtered to remove un-reacted solid. The clear solution was directly purified by reverse phase flash chromatography (C18 column, acetonitrile and water, neutral condition) to afford 36 (60 mg, yield 12%) as a white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 7.60 (dd, J=8.0, 3.8 Hz, 4H), 6.97 (dd, J=8.2, 3.8 Hz, 4H), 2.47 (s, 4H), 2.23 (q, J=6.6 Hz, 4H), 2.10 (d, J=12.9 Hz, 2H), 1.77-1.62 (m, 4H), 1.53-1.03 (m, 8H). LCMS: m/z calculated for C.sub.28H.sub.34I2N.sub.2O.sub.8Pt: 975.48; found: 976.57 [M+H].sup.+.

Example 37

N-((S)-1-(((R)-1-(6-(2-(2-(2-((S)-2-(4-isobutylphenyl)propanamido)ethoxy)ethoxy)ethyl)-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)-3-methylbutyl)amino)-1-oxo-3-phenylpropan-2-yl)pyra zine-2-carboxamide (37)

##STR00104##

##STR00105##

[0222] Step 1: To a solution of compound 37-1 (1.0 g, 4.02 mmol) in DMF (10 mL) was added NaHCO.sub.3 (3.38 g, 40.23 mmol), KI (669 mg, 4.03 mmol) and compound 9-2 (3.36 g, 20.11 mmol). The mixture was stirred at room temperature for 4 h. After completion of the reaction, the mixture was partitioned between EtOAc (100 mL) and water (100 mL). The organic layer was washed with brine, dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The residue was purified by silica gel column (eluted with 50% EtOAc in petroleum ether) to afford the title compound 37-2 (1.14 g, yield 67%). LCMS: m/z calculated for C.sub.19H.sub.36N.sub.2O.sub.8: 420.50; found: 443.85 [M+Na].sup.+.

[0223] Step 2: A mixture of compound 37-2 (1.14 g, 2.71 mmol) in 4 N HCl (EtOAc solution, 10 mL) was stirred at room temperature for 1.5 h. After completion of the reaction, the mixture was concentrated to afford the title compound 37-3 (1.36 g, crude), which was used at the next step without further purification.

[0224] Step 3: A solution of compound 37-3 (1.36 g, crude), compound 8-1 (670 mg, 3.24 mmol), DIPEA (525 mg, 4.06 mmol), HOBt (360 mg, 2.66 mmol) and EDCI (520 mg, 2.71 mmol) in DCM (10 mL) was stirred at room temperature for 16 h. After completion of the reaction, the mixture was diluted with DCM (100 mL), washed with water (100 mL2) and brine (100 mL). The organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The crude product was purified by reverse phase flash chromatography (C18 column, eluted with 65% acetonitrile in water, HCl condition). The desired components were lyophilized to afford the title compound 37-4 (948 mg, TFA salt, yield 64% over 2 steps) as a white solid. LCMS: m/z calculated for C.sub.27H.sub.44N.sub.2O.sub.7: 508.66; found: 531.73 [M+Na].sup.+.

[0225] Step 4: To a solution of compound 37-4 (945 mg, 1.73 mmol) in EtOH (9.5 mL) and H.sub.2O (3 mL) was added NaOH (297 mg, 7.45 mmol) at 0 C. The reaction was stirred at room temperature for 2 h. The mixture was concentrated and adjusted to pH=2 with 1 N HCl at 0 C. The aqueous mixture was purified by reverse phase flash chromatography (C18 column, eluted with 45% acetonitrile in water, HCl condition). The desired components were lyophilized to afford the title compound 37-5 (694 mg, yield 82%) as a white solid. LCMS: m/z calculated for C.sub.23H.sub.36N.sub.2O.sub.7: 452.55; found: 453.52 [M+H].sup.+.

[0226] Step 5: A mixture of compound 37-5 (100 mg, 0.204 mmol) and compound 9-7 (85 mg, 0.22 mmol) in toluene (3 mL) was stirred at 120 C. for 4 h. After completion of the reaction, the mixture was concentrated. The crude product was purified by pre-TLC (acetonitrile:dichloromethane=1:3) to afford 37 (97 mg, yield 59%) as a white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 9.05 (s, 1H), 8.92 (d, J=7.8 Hz, 1H), 8.84 (s, 1H), 8.71 (s, 1H), 7.95 (s, 1H), 7.37 (d, J=9.3 Hz, 1H), 7.29-7.12 (m, 7H), 7.03 (d, J=6.9 Hz, 2H), 4.66 (d, J=6.2 Hz, 1H), 4.28 (d, J=17.6 Hz, 1H), 4.02 (dd, J=32.9, 17.2 Hz, 2H), 3.68 (t, J=18.8 Hz, 3H), 3.46 (s, 3H), 3.39-3.25 (m, 4H), 3.16 (s, 2H), 3.09-3.00 (m, 2H), 2.37 (s, 2H), 1.76 (s, 1H), 1.55 (s, 1H), 1.41 (t, J=12.4 Hz, 1H), 1.27 (d, J=6.3 Hz, 3H), 1.23-1.16 (m, 1H), 0.82 (d, J=6.5 Hz, 12H). LCMS: m/z calculated for C.sub.42H.sub.57BN.sub.6O.sub.9: 800.76; found: 801.76 [M+H].sup.+.

Example 38

(S)-4,11-diethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3,4:6,7]indolizino[1,2-b]quinolin-9-yl(S)-2-(4-(2-(methyl-d3)propyl-3,3,3-d3)phenyl)propanoate (38)

##STR00106##

##STR00107##

[0227] Step 1: A mixture of compound 22-11 (30.0 mg, 0.141 mmol), EDCI (80.0 mg, 0.419 mmol) and HOBt (20.0 mg, 0.148 mmol) in anhydrous DMF (2 mL) was stirred at room temperature for 5 min. Compound 1-1 (85 mg, 0.216 mmol) was added and the mixture was stirred at room temperature for 16 h. The reaction mixture was partition between EtOAc (30 mL) and water (20 mL). The organic layer was washed with brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The crude product was purified by pre-TLC (5% MeOH in DCM) to afford the title compound 38 (50 mg, yield 60%) as an off-white solid. .sup.1H NMR (500 MHz, DMSO-d.sub.6) 8.18 (d, J=9.1 Hz, 1H), 7.90 (d, J=2.5 Hz, 1H), 7.51 (dd, J=9.1, 2.5 Hz, 1H), 7.37 (d, J=7.9 Hz, 2H), 7.32 (s, 1H), 7.20 (d, J=7.8 Hz, 2H), 5.43 (s, 2H), 5.32 (s, 2H), 4.15 (q, J=7.1 Hz, 1H), 3.16 (q, J=7.6 Hz, 2H), 2.45 (d, J=7.2 Hz, 2H), 1.85 (ddt, J=28.9, 14.6, 7.2 Hz, 3H), 1.57 (d, J=7.1 Hz, 3H), 1.27 (t, J=7.6 Hz, 3H), 0.88 (t, J=7.3 Hz, 3H). LCMS: m/z calculated for C.sub.35H.sub.30D.sub.6N.sub.2O.sub.6: 586.72; found: 588.13 [M+H].sup.+.

Example 39. Cellular IC50 in Three Tumor Cell Lines

[0228] The cellular inhibition was determined in three assays: U87MG, A549 and MC38. Cells were recovered and cultured in appropriate medium supplemented with 10% fetal bovine serum and 100 U/mL penicillin G sodium, and maintained in cell incubators (37 C., 5% CO2). Before testing, cells in culture dishes were rinsed with Phosphate Buffered Solution, detached with Trypsin. Dilute and adjust the cell number with the culture medium, add the cell suspension to the 96 well cell plate. Cells were maintained in incubators overnight. In T0 control plate, cells were added with 100 L CellTiter-Glo reagent, balance at room temperature for 10 minutes, and read the chemiluminescence value with envision.

[0229] In the testing plate, compounds were dissolved with corresponding solvent and diluted gradiently. Diluted compound solution was added to a 96 well plate with cells. After incubation of 72 h, cells were added with 100 L CellTiter-Glo reagent, balanced at room temperature for 10 minutes, and read the chemiluminescence value with envision. IC50 was calculated using the GraphPad Prism software package (Prism 6 for Windows, Version 6.0, GraphPad Software Inc., San Diego, CA). The IC.sub.50 for the three cellular assays is shown in Table 1.

TABLE-US-00004 TABLE 1 IC50 U87MG A549 MC38 Compound (IC50, nM) (IC50, nM) (IC50, nM) 8 13.37693 207.81632 22.74152 16 7.266483 10.68657 15.94872 9-7 (Valcade ) 6.890381 9.36019 16.29071 1-1 (SN-38) 13.58112 99.519661 21.24506 5-1 NA NA NA 8-1 #Intersect #Intersect >10000 16-2 #Intersect #Intersect #Intersect A: <1 M; B: 1-10 M. C: >10 M

Example 40. Chemical Stability in PBS (pH 7.4)

[0230] The chemical stability assay was performed according to the following procedures. [0231] 1. Test compounds spiking solution: 1 mM test compounds spiking solution A: Add 10 L of 10 mM test compounds stock solution to 90 L DMSO. [0232] 2. Add 396 L of buffer into the tubes designated for different time points. Pre-warm the samples at 37 C. for 10 min. [0233] 3. Add 4 L of spiking solution A into the wells designated as 0 min (or 15, 45, 90, 120 min) containing 396 L of buffer, then start timing count down. [0234] 4. At each time point, add 1200 L ACN containing IS into the tubes. [0235] 5. Samples are centrifuged at 10,000 rpm for 5 min, then 100 L of supernatant are ready for LC-MS/MS analysis.

TABLE-US-00005 TABLE 2 Stability Stability (PBS, pH Compound # 7.4) T (min) 16 8 4 215 30 806 31 1 1182 2 1387 3 51 28 504

Example 41. Pharmacokinetic

[0236] Pharmacokinetic parameters were determined in male CD-1 mice (Shanghai Jihui Laboratory Animal Care Co., Ltd., 6-8 weeks old). Animals were maintained under a 12 hr light/dark cycle. Animals had free access to food and water during the study.

[0237] For intravenous, compounds were formulated as a solution (in 5% DMSO+10% Solutol HS15+85% (20% HP--CD in water)) in 5 mL/kg dosing volume and administered via tail vein. For subcutaneous, compounds were formulated as a solution (in 5% DMSO+10% Solutol HS15+85% (20% HP--CD in water)) in 10 mL/kg dosing volume or (in 0.5% MC+1% Pluronic F68 in water) in 50 L/mouse dosing volume and administered via subcutaneous puncture.

[0238] Semi-serial blood samples (about 110 L) were taken from animal at 0.083, 0.25, 0.5, 1, 2, 4, 8 and 24 hr for IV group and 0.5, 2, 4, 8, 24, 48 and 96 hr for SC group. Samples were held on ice for no longer than 15 minutes before centrifugation (2000 g, 5 min, 4 C.) within 15 minutes post sampling. Plasma was snap frozen in dry ice and then transferred into 70 C. freezer for long term storage until LC-MS/MS analysis.

[0239] Tissue collection for SC group at 0.5, 2, 4, 8, 24, 48 and 96 hr. After animals were anesthetized and exsanguinated, tissue samples (including muscle and skin) were collected and weighted, and then snap frozen in liquid nitrogen and further stored at 70 C. for long term storage until LC-MS/MS analysis. Tissue samples were homogenized under freezing conditions.

[0240] PK parameters were generated from LC-MS/MS data using Phoenix WinNonlin 8.2 software.

TABLE-US-00006 TABLE 3 Pharmacokinetic Data Dosing Dose Compound AUClast AUClast Tissue/Plasma compound (mg/kg) measured T (h) (hr*ng/mL) (hr*ng/mL) ratio 1 3 1 active drug 10.3 24402 142 171.8450704 1-1 8 5 8 44.1 11834033 NA #VALUE! 8 5 8 active drug >96 255830 201 1272.78607 1-1 28 10 28 11.3 37582 NA #VALUE! 28 10 active drug >48 4094 NA #VALUE! 28-1 30 10 active drug 25.4 144212 5274 27.34395146 30-1 31 10 active drug 34.2 979 203 4.822660099 31-4 33 10 active drug NA 9139 504 18.13293651 33-1 16 1 16 93.2 60283 165 365.3515152 16 1 active drug >96 5776 13.5 427.8518519 9-7 10 1 active drug 22.4 5919 10.2 580.2941176 9-7 13 1 active drug 22 11243 284 39.58802817 9-7

[0241] Applicant's disclosure is described herein in preferred embodiments with reference to the Figures, in which like numbers represent the same or similar elements. Reference throughout this specification to one embodiment, an embodiment, or similar language means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one embodiment of the present invention. Thus, appearances of the phrases in one embodiment, in an embodiment, and similar language throughout this specification may, but do not necessarily, all refer to the same embodiment.

[0242] The described features, structures, or characteristics of Applicant's disclosure may be combined in any suitable manner in one or more embodiments. In the description, herein, numerous specific details are recited to provide a thorough understanding of embodiments of the invention. One skilled in the relevant art will recognize, however, that Applicant's composition and/or method may be practiced without one or more of the specific details, or with other methods, components, materials, and so forth. In other instances, well-known structures, materials, or operations are not shown or described in detail to avoid obscuring aspects of the disclosure.

[0243] In this specification and the appended claims, the singular forms a, an, and the include plural reference, unless the context clearly dictates otherwise.

[0244] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present disclosure, the preferred methods and materials are now described. Methods recited herein may be carried out in any order that is logically possible, in addition to a particular order disclosed.

INCORPORATION BY REFERENCE

[0245] References and citations to other documents, such as patents, patent applications, patent publications, journals, books, papers, web contents, have been made in this disclosure. All such documents are hereby incorporated herein by reference in their entirety for all purposes. Any material, or portion thereof, that is said to be incorporated by reference herein, but which conflicts with existing definitions, statements, or other disclosure material explicitly set forth herein is only incorporated to the extent that no conflict arises between that incorporated material and the present disclosure material. In the event of a conflict, the conflict is to be resolved in favor of the present disclosure as the preferred disclosure.

EQUIVALENTS

[0246] The representative examples are intended to help illustrate the invention, and are not intended to, nor should they be construed to, limit the scope of the invention. Indeed, various modifications of the invention and many further embodiments thereof, in addition to those shown and described herein, will become apparent to those skilled in the art from the full contents of this document, including the examples and the references to the scientific and patent literature included herein. The examples contain important additional information, exemplification and guidance that can be adapted to the practice of this invention in its various embodiments and equivalents thereof.