Boronic ester compounds and pharmaceutical compositions thereof

Abstract

The present invention provides novel compounds useful as proteasome inhibitors. The present invention also provides pharmaceutical compositions comprising the compounds of the present invention, and methods of using the compositions to treat various diseases.

Description

[0001] This application is a divisional application of application number 201310348813.5, filed on June 16, 2009, with the title "Boronate ester compounds and pharmaceutical compositions thereof".

[0002] Priority

[0003] This application claims priority to U.S. Provisional Patent Application No. 61 / 132,244, filed June 17, 2008, and U.S. Provisional Patent Application No. 61 / 211,499, filed March 31, 2009, the entire contents of each of which are incorporated herein by reference. TECHNICAL FIELD

[0004] The present invention relates to boronate ester compounds useful as proteasome inhibitors. The present invention also provides pharmaceutical compositions comprising the compounds of the present invention, and methods of using the compositions to treat various diseases. BACKGROUND

[0005] Boronic acid and boronate ester compounds exhibit a variety of biologically activities that are medically useful. U.S. Patent No. 4,499,082 (1985) to Shenvi et al. discloses peptide boronic acids as inhibitors of certain proteolytic enzymes. U.S. Patent Nos. 5,187,157 (1993), 5,242,904 (1993), and 5,250,720 (1993) to Kettner and Shenvi describe a class of peptide boronic acids that inhibit proteases like trypsin. U.S. Patent No. 5,169,841 (1992) to Kleeman et al. discloses N-terminally modified peptide boronic acids that inhibit the action of renin. U.S. Patent No. 5,106,948 (1992) to Kinder et al. discloses certain boronic acid compounds that inhibit the growth of cancer cells. WO 04 / 022070 to Magde et al. discloses peptide boronic acid compounds that inhibit thrombin. U.S. Patent Application Publication No. 2006 / 0084592 to Boucher discloses various base addition salts of peptide boronic acid compounds. WO 07 / 005991 to Bachovchin et al. discloses peptide boronic acid compounds that inhibit fibroblast activation protein.

[0006] Boric acid and boronate compounds are of particular interest as inhibitors of proteasomes, the multicatalytic proteases responsible for most intracellular protein turnover. U.S. Patent No. 5,780,454 to Adams et al. (1998) describes peptide boronates and boronic acid compounds useful as proteasome inhibitors. The reference also describes the use of boronate and boronic acid compounds to reduce the rate of muscle protein degradation, to reduce the activity of NF-κB in cells, to reduce the rate of degradation of p53 protein in cells, to inhibit cyclin degradation in cells, to inhibit the growth of cancer cells, and to inhibit NF-κB dependent cell adhesion. WO 02 / 096933 to Furet et al., WO 05 / 016859 to Chatterjee et al., and WO 05 / 021558 and WO 06 / 08660 to Bernadini et al. disclose other boronate and boronic acid compounds reportedly having proteasome inhibitory activity.

[0007] Ciechanover, Cell, 79: 13-21 (1994) discloses that proteasomes are the proteolytic component of the ubiquitin-proteasome pathway in which proteins are targeted for degradation by conjugation to multiple ubiquitin molecules. Ciechanover also discloses that the ubiquitin-proteasome pathway plays a key role in a variety of important physiological processes. Rivett et al., Biochem. J. 291 : 1 (1993) discloses that proteasomes exhibit trypsin peptidase, chymotrypsin peptidase, and peptidyl glutamyl peptidase activities. The catalytic core that makes up the 26S proteasome is the 20S proteasome. McCormack et al., Biochemistry 37: 7792 (1998) teaches that the 20S proteasome cleaves a variety of peptide substrates, including Suc-Leu-Leu-Val-Tyr-AMC, Z-Leu-Leu-Arg-AMC, and Z-Leu-Leu-Glu-2NA, where Suc is N-succinyl, AMC is 7-amino-4-methylcoumarin, and 2NA is 2-naphthylamine.

[0008] Proteasome inhibition represents an important new strategy for cancer therapy. King et al., Science 274: 1652-1659 (1996) describe the essential role of the ubiquitin-proteasome pathway in regulating the cell cycle, neoplastic growth, and metastasis. The authors teach that a number of key regulatory proteins, including cyclins and cyclin-dependent kinases p21 and p27 KIP1 ) are temporally degraded by the ubiquitin-proteasome pathway during the cell cycle. The ordered degradation of these proteins is necessary for the cell to progress through the cell cycle and undergo mitosis.

[0009] In addition, the ubiquitin-proteasome pathway is required for transcriptional regulation. Palombella et al., Cell, 78:773 (1994) teaches that activation of the transcription factor NF-κB is regulated by proteasome-mediated degradation of the inhibitory protein IκB. NF-κB, in turn, plays an important role in the regulation of genes involved in immune and inflammatory responses. Read et al., Immunity 2:493-506 (1995) teaches that the ubiquitin-proteasome pathway is required for the expression of cell adhesion molecules such as E-selectin, ICAM-1, and VCAM-1. Zetter, Seminars in Cancer Biology 4:219-229 (1993) teaches that cell adhesion molecules are involved in tumor metastasis and angiogenesis in vivo by directing the adhesion of tumor cells to blood vessels and extravasation from the blood vessels to distant tissue sites in the body. In addition, Beg and Baltimore, Science 274:782 (1996) teach that NF-κB is an anti-apoptotic control factor and that inhibition of NF-κB activation makes cells more sensitive to environmental stress and cytotoxic agents.

[0010] Proteasome inhibitors Proteasome inhibitors (bortezomib; N-2-pyrazinecarbonyl-L-phenylalanine-L-leucine boronic acid) was the first proteasome inhibitor to gain regulatory approval. Mitsiades et al., Current Drug Targets, 7: 1341 (2006) reviews the clinical studies that led to the approval of bortezomib for the treatment of patients with multiple myeloma who have received at least one prior therapy. Fisher et al., J. Clin. Oncol., 30:4867 (2006) describes an international multicenter phase II study demonstrating the activity of bortezomib in patients with relapsed or refractory mantle cell lymphoma. Ishii et al., Anti-Cancer Agents in Medicinal Chemistry, 7:359 (2007) and Roccaro et al., Curr. Pharm. Biotech., 7: 1341 (2006) discuss various molecular mechanisms that can contribute to the anti-tumor activity of bortezomib.

[0011] Structural analysis reported by Voges et al., Annu. Rev. Biochem., 68: 1015 (1999) revealed that the 20S proteasome comprises 28 subunits, of which the catalytic subunits βl, β2 and β5 are responsible for peptidyl glutamyl peptidase, trypsin peptidase and chymotrypsin peptidase activity, respectively. Rivett et al., Curr. Protein Pept. Sci., 5: 153 (2004) revealed that when the proteasome is exposed to certain cytokines, including IFN-γ and TNF-α, the βl, β2 and β5 subunits are replaced by alternative catalytic subunits βli, β2i and β5i to form a variant form of the proteasome known as the immunoproteasome.

[0012] Orlowski, Hematology (Am. Soc. Hematol. Educ. Program) 220 (2005) revealed that the immunoproteasome is also constitutively expressed in some cells derived from hematopoietic precursors. The authors suggest that inhibitors specific for the immunoproteasome can be useful for targeted therapy of blood-derived cancers, thereby potentially sparing normal tissues (e.g., gastrointestinal and neural tissues) from side effects.

[0013] Unfortunately, boronic acid compounds are relatively difficult to obtain in analytically pure form. For example, Snyder et al., J. Am. Chem. Soc. 80: 3611 (1958) teach that aryl boronic acid compounds are prone to form cyclic trimeric anhydrides under dehydrating conditions. Additionally, alkyl boronic acids and their boroxines are generally air sensitive. Korcek et al., J. Chem. Soc, Perkin Trans. 2 242 (1972) teach that butyl boronic acid is readily oxidized by air to form 1-butanol and boric acid. These difficulties limit the medical utility of boronic acid compounds, complicate the characterization of pharmaceutical agents comprising boronic acid compounds, and limit their shelf life.

[0014] Plamondon et al., WO 02 / 059131 disclose stable, pharmaceutically acceptable compositions prepared from boronic acid compounds and sugars. There remains a need for additional stable formulations of boronic acid compounds. SUMMARY

[0015] The present invention provides novel boronate compounds and stable, pharmaceutically acceptable compositions comprising the compounds. These compounds and compositions are useful for inhibiting proteasome activity in vitro and in vivo, and are particularly useful for treating various cell proliferative diseases.

[0016] The present invention relates to the following technical solutions:

[0017] 1. A compound of formula (I): (I)

[0018]

[0019] or a pharmaceutically acceptable salt thereof, wherein:

[0020] A is 0, 1 or 2;

[0021] P is hydrogen or an amino capping moiety;

[0022] R a is hydrogen, C 1-6 aliphatic, C 1-6 fluoroaliphatic, -(CH2) m -CH2-R B , -(CH2) m -CH2-NHC(=NR 4 )NH-Y, -(CH2) m -CH2-CON(R 4 )2, -(CH2) m -CH2-N(R 4 )CON(R 4 )2, -(CH2) m -CH(R 6 )N(R 4 )2, -(CH2) m -CH(R 5a )-OR 5b or -(CH2) m -CH(R 5 )-SR 5 ;

[0023] R a1 is hydrogen, C 1-6 aliphatic, C 1-6 fluoroaliphatic, -(CH2) m -CH2-R B , -(CH2) m -CH2-NHC(=NR 4 )NH-Y, -(CH2) m -CH2-CON(R 4 )2, -(CH2) m -CH2-N(R 4 )CON(R 4 )2, -(CH2) m -CH(R 6 )N(R 4 )2, -(CH2)m -CH(R 5a )-OR 5b or -(CH2) m -CH(R 5 )-SR 5 ;

[0024] each R a2 is independently hydrogen, C 1-6 aliphatic, C 1-6 fluoroaliphatic, -(CH2) m -CH2-R B , -(CH2) m -CH2-NHC(=NR 4 )NH-Y, -(CH2) m -CH2-CON(R 4 )2, -(CH2) m -CH2-N(R 4 )CON(R 4 )2, -(CH2) m -CH(R 6 )N(R 4 )2, -(CH2) m -CH(R 5a )-OR 5b or -(CH2) m -CH(R 5 )-SR 5 ;

[0025] each R B is independently a substituted or unsubstituted monocyclic or bicyclic ring system;

[0026] each R 4 is independently hydrogen or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl; or two R 4 on the same nitrogen atom are taken together to form a substituted or unsubstituted 4- to 8-membered heterocyclyl ring having, in addition to the nitrogen atom, 0-2 ring heteroatoms independently selected from N, O, and S;

[0027] each R 5 is independently hydrogen or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl;

[0028] each R 5a is independently hydrogen or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl;

[0029] each R 5b is independently hydrogen or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl;

[0030] each R 6 is independently substituted or unsubstituted aliphatic, aryl, or heteroaryl;

[0031] Y is hydrogen, -CN, or -NO2;

[0032] m is 0, 1, or 2; and

[0033] Z 1 forms, together with Z 2 a moiety derived from an alpha-hydroxy carboxylic acid, wherein the atom attached to boron in each instance is an oxygen atom; or Z 1 forms, together with Z 2 a moiety derived from a beta-hydroxy carboxylic acid, wherein the atom attached to boron in each instance is an oxygen atom.

[0034] 2. The compound according to item 1, characterized by formula (II):

[0035]

[0036] or a pharmaceutically acceptable salt thereof, wherein:

[0037] each R b1 and R b2 is independently hydrogen, -CO2H, -OH, or substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl;

[0038] each R b3 and R b4 is independently hydrogen, -CO2H, or substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl;

[0039] or R b2 and R b4 are each independently hydrogen, and R b1 and R b3 are taken together with the carbon atom to which they are attached to form an unsubstituted or substituted fused 4- to 8-membered non-aromatic ring having 0-3 ring heteroatoms selected from the group consisting of O, N, and S, wherein the ring can optionally be fused to an unsubstituted or substituted 4- to 8-membered non-aromatic ring or 5- to 6-membered aromatic ring having 0-3 ring heteroatoms selected from the group consisting of O, N, and S;

[0040] or R b2 and R b4 are absent, and R b1 and R b3The ring is bonded to the carbon atom to form an unsubstituted or substituted fused 5- to 6-membered aromatic ring having 0-3 cyclic heteroatoms selected from the groups of O, N and S, wherein the ring may optionally be fused with an unsubstituted or substituted 4- to 8-membered nonaromatic ring or a 5- to 6-membered aromatic ring having 0-3 cyclic heteroatoms selected from the groups of O, N and S; and n is 0 or 1.

[0041] 3. The compound according to claim 2, wherein R b3 and R b4 Each of them is independently hydrogen, C 1-6 Aliphatic or -(CH2) p -CO2H;

[0042] And p is 0, 1 or 2.

[0043] 4. The compound according to item 3, wherein P is R c -C(O)-、R c -OC(O)-、R c -N(R 4c )-C(O)-、R c -S(O)2- or R c -N(R 4c )-S(O)2-;

[0044] R c Choose C freely 1-6 aliphatic, C 1-6 Fluoroaliphatic, -R D -T 1 -R D and -T 1 -R 2c Groups formed;

[0045] T 1 For 0-2 independently selected R 3a or R 3b Replacement C 1-6 Alkylene chains, wherein the alkylene chains are optionally doped with -C(R) 5 )=C(R 5 -, -C≡C- or -O-;

[0046] R D For substituted or unsubstituted single-ring or double-ring systems;

[0047] R 2c Halogenated, -OR 5 -SR 6 -S(O)R 6 -SO2R 6 -SO2N(R)4 )2、-N(R 4 )2、-NR 4 C(O)R 5 -NR 4 C(O)N(R 4 )2、-NR 4 CO2R 6 -N(R) 4 SO2R 6 -N(R) 4 SO2N(R) 4 )2、-OC(O)R 5 -OC(O)N(R) 4 )2、-C(O)R 5 -CO2R 5 or -C(O)N(R) 4 )2;

[0048] Each R 3a Independently selectable from -F, -OH, -O(C) 1-4 Alkyl), -CN, -N(R) 4 )2、-C(O)(C 1-4 Alkyl group, -CO2H, -CO2(C 1-4 Alkyl groups, -C(O)NH2 and -C(O)-NH(C 1-4 A group consisting of alkyl groups;

[0049] Each R 3b Independently for R 3a or R 7 C, whether substituted or not 1-3 Aliphatic; or two substituents R on the same carbon atom 3b It combines with the carbon atoms it is attached to to form 3- to 6-membered alicyclic rings;

[0050] Each R 7 It is a substituted or unsubstituted aromatic group; and

[0051] R 4c For hydrogen, C 1-4 Alkyl, C 1-4 fluoroalkyl or C 6-10 Fang (C) 1-4 )alkyl, wherein the aryl moiety is substituted or unsubstituted.

[0052] 5. The compound according to item 3, wherein A is 0;

[0053] R a1 It is hydrogen, -(CH2) m -CH2-R B Or -(CH2)m -CH(R 5a )-OR 5b ; and

[0054] m is 0.

[0055] 6. The compound according to item 3, wherein P is R c -C(O)- or R c -S(O)2- and R c is -R D .

[0056] 7. The compound according to item 6, wherein R D is a substituted or unsubstituted monocyclic or bicyclic ring system selected from the group consisting of phenyl, pyridyl, pyrimidinyl, pyrazinyl, naphthyl, benzimidazolyl, quinolinyl, isoquinolinyl, quinoxalinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, tetrahydroquinoxalinyl, and dihydrobenzoxazinyl.

[0057] 8. The compound according to item 7, wherein R D is substituted on the substitutable carbon atoms with 0-1 R d and 0-2 R 8d ;

[0058] each R d is independently C 1-6 aliphatic, C 1-6 fluoroaliphatic, or halo; and

[0059] each R 8d is independently C 1-4 aliphatic, C 1-4 fluoroaliphatic, or halo.

[0060] 9. The compound according to item 4, wherein A is 0;

[0061] R a is C 1-6 aliphatic or -(CH2) m -CH2-R B ;

[0062] R a1 is hydrogen, -(CH2) m -CH2-R B , or -(CH2) m -CH(R 5a )-OR 5b ;

[0063] P is R c -C(O)- or R c -S(O)2-;

[0064] R c is -R D ; and

[0065] m is 0 or 1.

[0066] 10. The compound according to item 9, wherein R a is C 1-6 aliphatic.

[0067] 11. The compound according to item 10, wherein R a1 is -CH2-R B ;

[0068] R B is phenyl; and

[0069] R D is 2-pyrazinyl.

[0070] 12. The compound according to item 10, wherein R a1 is -CH(R 5a )-OR 5b ;

[0071] R 5a is C 1-6 aliphatic;

[0072] R 5b is hydrogen; and

[0073] R D is 6-phenyl-2-pyridinyl-.

[0074] 13. The compound according to item 10, wherein R a1 is hydrogen, and

[0075] R D is 2,5-dichlorophenyl.

[0076] 14. The compound according to item 3, wherein Z 1 together with Z 2 forms a moiety derived from citric acid.

[0077] 15. The compound according to item 14, in a substantially crystalline form.

[0078] 16. The compound according to item 14, characterized by the formula (III), (Ilia), (IV), (IVa):

[0079]

[0080] or mixtures thereof.

[0081] 17. The compound according to item 16, wherein R a is C 1-6 aliphatic.

[0082] 18. The compound according to item 16, wherein A is 0; R a1 is hydrogen, -(CH2) m -CH2-R B or -(CH2) m -CH(R 5 )-OR 5b ; P is R c -C(O)-; and R c is -R D .

[0083] 19. The compound according to item 18, wherein R D is a substituted or unsubstituted monocyclic or bicyclic ring system selected from the group consisting of phenyl, pyridyl, pyrimidinyl, pyrazinyl, naphthyl, benzimidazolyl, quinolinyl, isoquinolinyl, quinoxalinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, tetrahydroquinoxalinyl, and dihydrobenzoxazinyl.

[0084] 20. The compound according to item 18, wherein R a is isobutyl.

[0085] 21. The compound according to item 20, wherein R a1 is -CH2-R B ;

[0086] R B is phenyl; and

[0087] R D is 2-pyrazinyl.

[0088] 22. The compound according to item 20, wherein R a1 is hydrogen, and

[0089] R D is 2,5-dichlorophenyl.

[0090] 23. The compound according to item 20, wherein R a1 is -CH(R 5a )-OR 5b ;

[0091] R 5a is C 1-6 aliphatic;

[0092] R 5b is hydrogen; and

[0093] R DR is hydrogen, C1-6 aliphatic, C1-6 haloaliphatic, -(CH2)0-6-phenyl, -(CH2)0-6-5- to 7-membered heterocyclyl, -(CH2)0-6-5- to 7-membered heteroaryl, -C(=O)R, -C(=O)OR, -C(=O)N(R)2, -OR, -SR, -N(R)2, -N(R)C(=O)R, -OC(O)N(R)2, -N(R)N(R)2, -N(R)OR, -CN, -OCN, -SCN, -NCO, -OC(O)R, -N(R)C(O)R, -OC(O)OR, -N(R)S(O)tR (t is 1 or 2), -OS(O)tR (t is 1 or 2), -S(O)tR (t is 1 or 2), -S(O)tN(R)2 (t is 1 or 2), -S(O)tOR (t is 1 or 2), -OP(O)2OR, -P(O)2N(R)2, -OP(O)2N(R)2, -Si(R)3, -OSi(R)3, -S(O)tR (t is 1 or 2), -Si(O)tR (t is 1 or 2), -Si(O)tN(R)2 (t is 1 or 2), -Si(O)tOR (t is 1 or 2), -N(R)P(O)2OR, -N(R)P(O)2N(R)2, -N(R)Si(R)3, -N(R)OSi(R)3, -N(R)S(O)tR (t is 1 or 2), -N(R)S(O)tN(R)2 (t is 1 or 2), -N(R)S(O)tOR (t is 1 or 2), -OP(O)2N(R)2, -OP(O)2R, -Si(O)tN(R)2 (t is 1 or 2), -Si(O)tR (t is 1 or 2), or -Si(O)tOR (t is 1 or 2); or a crystalline form thereof, a filler, and optionally a lubricant;

[0094] 24. A pharmaceutical composition comprising a compound of Formula (I):

[0095]

[0096] or a crystalline form thereof, a filler, and optionally a lubricant;

[0097] wherein:

[0098] A is 0, 1, or 2;

[0099] P is hydrogen or an amino capping moiety;

[0100] R a is hydrogen, C 1-6 1-6 aliphatic, C 1-6 1-6 haloaliphatic, -(CH2) m -CH2-R B , -(CH2) m -CH2-NHC(=NR 4 )NH-Y, -(CH2) m -CH2-CON(R 4 )2, -(CH2) m -CH2-N(R 4 )CON(R 4 )2, -(CH2) m -CH(R 6 )N(R 4 )2, -(CH2) m -CH(R 5a )-OR 5b , or -(CH2) m -CH(R 5 )-SR 5 ;

[0101] R a1 is hydrogen, C 1-6 1-6 aliphatic, C 1-6 1-6 haloaliphatic, -(CH2) m -CH2-R B , -(CH2) m -CH2-NHC(=NR 4 )NH-Y, -(CH2) m -CH2-CON(R 4 )2, -(CH2) m -CH2-N(R 4 )CON(R 4 )2, -(CH2) m -CH(R 6N(R 4 )2, -(CH2) m -CH(R 5a )-OR 5b or -(CH2) m -CH(R 5 )-SR 5 ;

[0102] each R a2 is independently hydrogen, C 1-6 aliphatic, C 1-6 fluoroaliphatic, -(CH2) m -CH2-R B , -(CH2) m -CH2-NHC(=NR 4 )NH-Y, -(CH2) m -CH2-CON(R 4 )2, -(CH2) m -CH2-N(R 4 )CON(R 4 )2, -(CH2) m -CH(R 6 )N(R 4 )2, -(CH2) m -CH(R 5a )-OR 5b or -(CH2) m -CH(R 5 )-SR 5 ;

[0103] each R B is independently a substituted or unsubstituted monocyclic or bicyclic ring system;

[0104] each R 4 is independently hydrogen or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl; or two R 4 on the same nitrogen atom are taken together to form a substituted or unsubstituted 4- to 8-membered heterocyclyl ring having, in addition to the nitrogen atom, 0-2 ring heteroatoms independently selected from N, O, and S;

[0105] each R 5 is independently hydrogen or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl;

[0106] each R 5a is independently hydrogen or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl;

[0107] each R 5bindependently hydrogen or substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl;

[0108] each R 6 independently substituted or unsubstituted aliphatic, aryl, or heteroaryl;

[0109] Y is hydrogen, -CN, or -NO2;

[0110] m is 0, 1, or 2; and

[0111] Z 1 together with Z 2 form a moiety derived from an alpha-hydroxy carboxylic acid, wherein the atom attached to boron in each instance is an oxygen atom; or Z 1 together with Z 2 form a moiety derived from a beta-hydroxy carboxylic acid, wherein the atom attached to boron in each instance is an oxygen atom.

[0112] 25. The pharmaceutical composition of item 24, wherein the pharmaceutical composition optionally further comprises a glidant, and optionally further comprises a buffer.

[0113] 26. The pharmaceutical composition of item 25, wherein the pharmaceutical composition comprises, in weight percent of total weight, about 0.2% to about 3% of the compound of formula (I) or a crystalline form thereof, about 86.5% to about 99.8% of a filler, optionally up to about 1.5% of a lubricant, optionally up to about 5% of a glidant, and optionally up to about 5% of a buffer.

[0114] 27. The pharmaceutical composition of item 25, wherein the pharmaceutical composition comprises, in weight percent of total weight, about 0.2% to about 3% of the compound of formula (I) or a crystalline form thereof, about 97% to about 99.8% of a filler, and optionally up to about 1.5% of a lubricant.

[0115] 28. The pharmaceutical composition of item 25, wherein the pharmaceutical composition comprises, in weight percent of total weight, about 0.25% to about 2% of the compound of formula (I) or a crystalline form thereof, and about 98% to about 99.75% of a filler.

[0116] 29. The pharmaceutical composition of item 25, wherein the pharmaceutical composition is an oral pharmaceutical dosage form.

[0117] 30. The pharmaceutical composition of item 29, wherein the oral pharmaceutical dosage form is a capsule.

[0118] 31. The pharmaceutical composition of item 25, wherein the compound of Formula (I), or a crystalline form thereof, is present in an amount of about 0.2% to about 3% by weight based on total weight.

[0119] 32. The pharmaceutical composition of item 25, wherein the compound of Formula (I), or a crystalline form thereof, is present in an amount of about 0.25% to about 2% by weight based on total weight.

[0120] 33. The pharmaceutical composition of item 25, wherein the filler is present in an amount of about 97% to about 99.8% by weight based on total weight.

[0121] 34. The pharmaceutical composition of item 25, wherein the filler is present in an amount of about 98% to about 99.75% by weight based on total weight.

[0122] 35. The pharmaceutical composition of item 25, wherein the filler is present in an amount of about 86.5% to about 99.8% by weight based on total weight.

[0123] 36. The pharmaceutical composition of item 25, wherein the filler is selected from the group consisting of powdered cellulose, microcrystalline cellulose, silicified microcrystalline cellulose, low moisture microcrystalline cellulose, sodium starch glycolate, pregelatinized starch, and mixtures thereof.

[0124] 37. The pharmaceutical composition of item 25, wherein the filler is selected from the group consisting of low moisture microcrystalline cellulose, sodium starch glycolate, pregelatinized starch, and mixtures thereof.

[0125] 38. The pharmaceutical composition of item 25, wherein the lubricant, when present, is present in an amount of up to about 1.5% by weight based on total weight.

[0126] 39. The pharmaceutical composition of item 25, wherein the lubricant, when present, is present in an amount of about 1% by weight based on total weight.

[0127] 40. The pharmaceutical composition of item 25, wherein the lubricant, when present, is selected from the group consisting of magnesium stearate, glyceryl behenate, hydrogenated vegetable oil, talc, zinc stearate, calcium stearate, sucrose stearate, sodium stearyl fumarate, and mixtures thereof.

[0128] 41. The pharmaceutical composition of item 25, wherein the lubricant is magnesium stearate.

[0129] 42. The pharmaceutical composition of item 25, wherein the glidant, when present, is present in an amount of up to about 5% by weight based on the total weight.

[0130] 43. The pharmaceutical composition of item 25, wherein the glidant, when present, is present in an amount of about 1% by weight based on the total weight.

[0131] 44. The pharmaceutical composition of item 25, wherein the glidant, when present, is talc.

[0132] 45. The pharmaceutical composition of item 25, wherein the buffering agent, when present, is present in an amount of up to about 5% by weight based on the total weight.

[0133] 46. The pharmaceutical composition of item 25, wherein the buffering agent, when present, is present in an amount of about 2% by weight based on the total weight.

[0134] 47. The pharmaceutical composition of item 25, wherein the buffering agent, when present, is sodium citrate.

[0135] 48. The pharmaceutical composition of item 25, wherein:

[0136] the α-hydroxy carboxylic acid or the β-hydroxy carboxylic acid is citric acid;

[0137] A is 0;

[0138] R a is isobutyl;

[0139] R a1 is hydrogen, C 1-6 aliphatic, -(CH2) m -CH2-R B or -(CH2) m -CH(R 5a )-OR 5b ;

[0140] P is R c -C(O)-;

[0141] R c is -R D ;

[0142] m is 0 or 1 ;

[0143] the filler is selected from the group consisting of low moisture microcrystalline cellulose, sodium starch glycolate, pregelatinized starch, and mixtures thereof; and

[0144] the lubricant, when present, is magnesium stearate.

[0145] 49. The pharmaceutical composition of clause 25, wherein

[0146] said compound of formula (I) is represented by compound (I-1), (I-15), or (I-18):

[0147]

[0148]

[0149] said filler is selected from the group consisting of low moisture microcrystalline cellulose, sodium starch glycolate, pregelatinized starch, and mixtures thereof; and

[0150] said lubricant, when present, is magnesium stearate.

[0151] 50. The pharmaceutical composition of clause 25, wherein:

[0152] said alpha-hydroxy carboxylic acid or said beta-hydroxy carboxylic acid is citric acid;

[0153] A is 0;

[0154] R a is isobutyl;

[0155] R a1 is hydrogen, C 1-6 aliphatic, -(CH2) m -CH2-R B or -(CH2) m -CH(R 5a )-OR 5b ;

[0156] P is R c -C(O)-;

[0157] R c is -R D ;

[0158] m is 0 or 1;

[0159] said filler is selected from the group consisting of low moisture microcrystalline cellulose, sodium starch glycolate, pregelatinized starch, and mixtures thereof;

[0160] said lubricant, when present, is magnesium stearate;

[0161] said glidant, when present, is talc; and

[0162] said buffering agent, when present, is sodium citrate.

[0163] 51. The pharmaceutical composition of clause 25, wherein

[0164] The compound of Formula (I) is represented by compound (I-1), (I-15), or (I-18):

[0165]

[0166]

[0167] The filler is selected from the group consisting of low moisture microcrystalline cellulose, sodium starch glycolate, pregelatinized starch, and mixtures thereof;

[0168] The lubricant, when present, is magnesium stearate;

[0169] The glidant, when present, is talc; and

[0170] The buffer, when present, is sodium citrate.

[0171] 52. A pharmaceutical composition comprising a compound of Formula (I-1):

[0172]

[0173] or a crystalline form thereof, a filler, and optionally a lubricant.

[0174] 53. The pharmaceutical composition of item 52, wherein:

[0175] The filler is selected from the group consisting of low moisture microcrystalline cellulose, sodium starch glycolate, pregelatinized starch, and mixtures thereof; and

[0176] The lubricant, when present, is magnesium stearate.

[0177] 54. The pharmaceutical composition of item 52, wherein the crystalline form is Form 2.

[0178] 55. The pharmaceutical composition of item 52, wherein the pharmaceutical composition optionally further comprises a glidant, and optionally further comprises a buffer.

[0179] 56. The pharmaceutical composition of item 55, wherein:

[0180] The filler is selected from the group consisting of low moisture microcrystalline cellulose, sodium starch glycolate, pregelatinized starch, and mixtures thereof;

[0181] The lubricant, when present, is magnesium stearate.

[0182] The glidant, when present, is talc; and

[0183] The buffer, when present, is sodium citrate.

[0184] 57. A unit dose pharmaceutical composition comprising a compound of Formula (I-1) or a crystalline form thereof, wherein the compound of Formula (I-1) is present in an amount that is equivalent in molar weight to about 0.1 mg to about 3.0 mg of the compound of Formula (VIII-1).

[0185] 58. A unit dose pharmaceutical composition comprising, by weight ratio, about 0.143 mg to about 4.3 mg of a compound of Formula (I-1) or a crystalline form thereof, measured as about 0.1 mg to about 3.0 mg of the compound of Formula (VIII-1).

[0186] 59. A pharmaceutical composition comprising a compound of Formula (I):

[0187]

[0188] or a crystalline form thereof, a bulking agent, and a buffer;

[0189] wherein:

[0190] A is 0, 1, or 2;

[0191] P is hydrogen or an amino capping moiety;

[0192] R a is hydrogen, C 1-6 aliphatic, C 1-6 fluoroaliphatic, -(CH2) m -CH2-R B , -(CH2) m -CH2-NHC(=NR 4 )NH-Y, -(CH2) m -CH2-CON(R 4 )2, -(CH2) m -CH2-N(R 4 )CON(R 4 )2, -(CH2) m -CH(R 6 )N(R 4 )2, -(CH2) m -CH(R 5a )-OR 5b , or -(CH2) m -CH(R 5 )-SR 5 ;

[0193] R a1 is hydrogen, C 1-6 aliphatic, C 1-6 fluoroaliphatic, -(CH2) m -CH2-R B , -(CH2)m -CH2-NHC(=NR 4 )NH-Y, -(CH2) m -CH2-CON(R 4 )2, -(CH2) m -CH2-N(R 4 )CON(R 4 )2, -(CH2) m -CH(R 6 )N(R 4 )2, -(CH2) m -CH(R 5a )-OR 5b , or -(CH2) m -CH(R 5 )-SR 5 ;

[0194] each R a2 is independently hydrogen, C 1-6 aliphatic, C 1-6 fluoroaliphatic, -(CH2) m -CH2-R B , -(CH2) m -CH2-NHC(=NR 4 )NH-Y, -(CH2) m -CH2-CON(R 4 )2, -(CH2) m -CH2-N(R 4 )CON(R 4 )2, -(CH2) m -CH(R 6 )N(R 4 )2, -(CH2) m -CH(R 5a )-OR 5b , or -(CH2) m -CH(R 5 )-SR 5 ;

[0195] each R B is independently a substituted or unsubstituted monocyclic or bicyclic ring system;

[0196] each R 4 is independently hydrogen or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl; or two R 4 on the same nitrogen atom are taken together to form a substituted or unsubstituted 4- to 8-membered heterocyclyl ring having, in addition to the nitrogen atom, 0-2 ring heteroatoms independently selected from N, O, and S;

[0197] each R 5 is independently hydrogen or substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl;

[0198] each R 5a is independently hydrogen or substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl;

[0199] each R 5b is independently hydrogen or substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl;

[0200] each R 6 is independently substituted or unsubstituted aliphatic, aryl, or heteroaryl;

[0201] Y is hydrogen, -CN, or -NO2;

[0202] m is 0, 1, or 2; and

[0203] Z 1 forms, together with Z 2 , a moiety derived from an alpha-hydroxy carboxylic acid, wherein the atom attached to boron in each instance is an oxygen atom; or Z 1 forms, together with Z 2 , a moiety derived from a beta-hydroxy carboxylic acid, wherein the atom attached to boron in each instance is an oxygen atom.

[0204] 60. The pharmaceutical composition of item 59, wherein the swelling agent is present in an amount of about 1% w / v to about 5% w / v.

[0205] 61. The pharmaceutical composition of item 59, wherein the swelling agent is present in an amount of about 3% w / v.

[0206] 62. The pharmaceutical composition of item 59, wherein the swelling agent is glycine.

[0207] 63. The pharmaceutical composition of item 59, wherein the buffering agent is sodium citrate and citric acid.

[0208] 64. The pharmaceutical composition of item 59, wherein

[0209] the alpha-hydroxy carboxylic acid or the beta-hydroxy carboxylic acid is citric acid;

[0210] A is 0;

[0211] R a is isobutyl;

[0212] R a1 is hydrogen, C 1-6aliphatic, -(CH2) m -CH2-R B or -(CH2) m -CH(R 5a )-OR 5b ;

[0213] P is R c -C(O)-;

[0214] R c is -R D ;

[0215] m is 0 or 1;

[0216] the bulking agent is glycine; and

[0217] the buffering agent is sodium citrate and citric acid.

[0218] 65. The pharmaceutical composition of clause 59, wherein

[0219] the compound of formula (I) is represented by compound (I-1), (I-15), or (I-18):

[0220]

[0221] the bulking agent is glycine; and

[0222] the buffering agent is sodium citrate and citric acid.

[0223] 66. A pharmaceutical composition comprising a compound of formula (I) and in the form of a lyophilized powder:

[0224]

[0225] wherein:

[0226] A is 0, 1, or 2;

[0227] P is hydrogen or an amino capping moiety;

[0228] R a is hydrogen, C 1-6 aliphatic, C 1-6 fluoroaliphatic, -(CH2) m -CH2-R B , -(CH2) m -CH2-NHC(=NR 4 )NH-Y, -(CH2) m -CH2-CON(R 4 )2, -(CH2) m -CH2-N(R 4 )CON(R4 )2, -(CH2) m -CH(R 6 )2, -(CH2) 4 )2, -(CH2) m -CH(R 5a )-OR 5b or -(CH2) m -CH(R 5 )-SR 5 ;

[0229] R a1 is hydrogen, C 1-6 aliphatic, C 1-6 fluoroaliphatic, -(CH2) m -CH2-R B , -(CH2) m -CH2-NHC(=NR 4 )NH-Y, -(CH2) m -CH2-CON(R 4 )2, -(CH2) m -CH2-N(R 4 )CON(R 4 )2, -(CH2) m -CH(R 6 )N(R 4 )2, -(CH2) m -CH(R 5a )-OR 5b or -(CH2) m -CH(R 5 )-SR 5 ;

[0230] each R a2 is independently hydrogen, C 1-6 aliphatic, C 1-6 fluoroaliphatic, -(CH2) m -CH2-R B , -(CH2) m -CH2-NHC(=NR 4 )NH-Y, -(CH2) m -CH2-CON(R 4 )2, -(CH2) m -CH2-N(R 4 )CON(R 4 )2, -(CH2) m -CH(R 6 )N(R 4 )2, -(CH2) m -CH(R 5a)-OR 5b or -(CH2) m -CH(R 5 )-SR 5 ;

[0231] each R B is independently a substituted or unsubstituted monocyclic or bicyclic ring system;

[0232] each R 4 is independently hydrogen or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl; or two R 4 on the same nitrogen atom are taken together to form a substituted or unsubstituted 4- to 8-membered heterocyclyl ring having, in addition to the nitrogen atom, 0-2 ring heteroatoms independently selected from N, O, and S;

[0233] each R 5 is independently hydrogen or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl;

[0234] each R 5a is independently hydrogen or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl;

[0235] each R 5b is independently hydrogen or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl;

[0236] each R 6 is independently a substituted or unsubstituted aliphatic, aryl, or heteroaryl;

[0237] Y is hydrogen, -CN, or -NO2;

[0238] m is 0, 1, or 2; and

[0239] Z 1 together with Z 2 forms a moiety derived from an alpha-hydroxycarboxylic acid, wherein the atom attached to boron in each instance is an oxygen atom; or Z 1 and Z 2 together form a moiety derived from a beta-hydroxycarboxylic acid, wherein the atom attached to boron in each instance is an oxygen atom.

[0240] 67. The pharmaceutical composition of clause 66, further comprising a bulking agent and a buffering agent.

[0241] 68. The pharmaceutical composition of clause 67, wherein the bulking agent is glycine.

[0242] 69. The pharmaceutical composition of clause 67, wherein the buffering agent is sodium citrate and citric acid.

[0243] 70. The pharmaceutical composition of clause 67, wherein

[0244] the a-hydroxy carboxylic acid or the β-hydroxy carboxylic acid is citric acid;

[0245] A is 0;

[0246] R a is isobutyl;

[0247] R a1 is hydrogen, C 1-6 aliphatic, -(CH2) m -CH2-R B or -(CH2) m -CH(R 5a )-OR 5b ;

[0248] P is R c -C(O)-;

[0249] R c is -R D ;

[0250] m is 0 or 1;

[0251] the bulking agent is glycine; and

[0252] the buffering agent is sodium citrate and citric acid.

[0253] 71. The pharmaceutical composition of clause 67, wherein

[0254] the compound of Formula (I) is represented by compound (I-l), (I-15), or (I-18):

[0255]

[0256] the bulking agent is glycine; and

[0257] the buffering agent is sodium citrate and citric acid.

[0258] 72. A unit dose pharmaceutical composition comprising a compound of Formula (I-l) and in the form of a lyophilized powder:

[0259]

[0260] wherein:

[0261] the compound of Formula (I-l) is present in an amount that is about the equivalent molar weight of about 1 mg to about 5 mg of the compound of Formula (VIII-l).

[0262] 73. The unit dose pharmaceutical composition of item 72, wherein the compound of Formula (I-l) is present in an amount equivalent to about 3.5 mg of the compound of Formula (VIII-l).

[0263] 74. The unit dose pharmaceutical composition of item 72, wherein the pharmaceutical composition further comprises glycine, sodium citrate, and citric acid.

[0264] 75. The unit dose pharmaceutical composition of item 74, wherein glycine is present in an amount of about 0.01 g to about 0.50 g.

[0265] 76. The unit dose pharmaceutical composition of item 74, wherein the sodium citrate and citric acid are present in an amount equivalent to about 0.005 g to about 0.250 g of citrate ion.

[0266] 77. A unit dose pharmaceutical composition comprising a compound of Formula (I- 15) and in the form of a lyophilized powder:

[0267]

[0268] wherein:

[0269] the compound of Formula (I-15) is present in an amount equivalent to about 1 mg to about 5 mg of the compound of Formula (VIII-15).

[0270] 78. The unit dose pharmaceutical composition of item 77, wherein the compound of Formula (I-15) is present in an amount equivalent to about 3.5 mg of the compound of Formula (VIII-15).

[0271] 79. The unit dose pharmaceutical composition of item 78, wherein the pharmaceutical composition further comprises glycine, sodium citrate, and citric acid.

[0272] 80. The unit dose pharmaceutical composition of item 78, wherein glycine is present in an amount of about 0.01 g to about 0.50 g.

[0273] 81. The unit dose pharmaceutical composition of item 78, wherein the sodium citrate and citric acid are present in an amount equivalent to about 0.005 g to about 0.250 g of citrate ion.

[0274] 82. A method of preparing a pharmaceutical composition of compound (I-l) as a lyophilized powder, the method comprising the steps of:

[0275] (f-1) combining the following to form a mixture:

[0276] i. water;

[0277] ii. the compound (I-1);

[0278] iii. glycine;

[0279] iv. sodium citrate; and

[0280] v. citric acid; and

[0281] (f-2) lyophilizing the mixture.

[0282] 83. A method of preparing a pharmaceutical composition of compound (I-15) as a lyophilized powder, the method comprising the steps of:

[0283] (g-1) combining the following to form a mixture:

[0284] i. an aqueous solvent mixture comprising water and tert-butanol;

[0285] ii. the compound (VIII-15);

[0286] iii. glycine;

[0287] iv. sodium citrate; and

[0288] v. citric acid; and

[0289] (g-2) lyophilizing the mixture.

[0290] 84. The method of item 83, wherein the tert-butanol in the aqueous solvent mixture is present in an amount of about 3% v / v to about 6% v / v tert-butanol.

[0291] 85. A method of preparing a pharmaceutical composition of compound (I-1) as a liquid pharmaceutical dosage form, the method comprising the step of reconstituting a lyophilized powder of compound (I-1) according to item 72 with water for injection.

[0292] 86. A method of preparing a pharmaceutical composition of compound (I-15) as a liquid pharmaceutical dosage form, the method comprising the step of reconstituting a lyophilized powder of compound (I-15) according to item 77 with water for injection.

[0293] 87. A liquid pharmaceutical composition comprising a compound of formula (I):

[0294]

[0295] a buffer and, optionally, a tonicity adjusting agent;

[0296] wherein:

[0297] A is 0, 1, or 2;

[0298] P is hydrogen or an amino capping moiety;

[0299] R a is hydrogen, C 1-6 aliphatic, C 1-6 fluoroaliphatic, -(CH2) m -CH2-R B , -(CH2) m -CH2-NHC(=NR 4 )NH-Y, -(CH2) m -CH2-CON(R 4 )2, -(CH2) m -CH2-N(R 4 )CON(R 4 )2, -(CH2) m -CH(R 6 )N(R 4 )2, -(CH2) m -CH(R 5a )-OR 5b or -(CH2) m -CH(R 5 )-SR 5 ;

[0300] R a1 is hydrogen, C 1-6 aliphatic, C 1-6 fluoroaliphatic, -(CH2) m -CH2-R B , -(CH2) m -CH2-NHC(=NR 4 )NH-Y, -(CH2) m -CH2-CON(R 4 )2, -(CH2) m -CH2-N(R 4 )CON(R 4 )2, -(CH2) m -CH(R 6 )N(R 4 )2, -(CH2) m -CH(R 5a )-OR 5b or -(CH2) m -CH(R 5 )-SR 5 ;

[0301] each R a2 is independently hydrogen, C 1-6 aliphatic, C 1-6 fluoroaliphatic, -(CH2) m-CH2-R B -(CH2) m -CH2-NHC(=NR 4 )NH-Y, -(CH2) m -CH2-CON(R 4 )2, -(CH2) m -CH2-N(R 4 )CON(R 4 )2, -(CH2) m -CH(R 6 )N(R 4 )2, -(CH2) m -CH(R 5a )-OR 5b , or -(CH2) m -CH(R 5 )-SR 5 ;

[0302] each R B is independently a substituted or unsubstituted monocyclic or bicyclic ring system;

[0303] each R 4 is independently hydrogen or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl; or two R 4 on the same nitrogen atom are taken together to form a substituted or unsubstituted 4- to 8-membered heterocyclyl ring having, in addition to the nitrogen atom, 0-2 ring heteroatoms independently selected from N, O, and S;

[0304] each R 5 is independently hydrogen or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl;

[0305] each R 5a is independently hydrogen or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl;

[0306] each R 5b is independently hydrogen or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl;

[0307] each R 6 is independently a substituted or unsubstituted aliphatic, aryl, or heteroaryl;

[0308] Y is hydrogen, -CN, or -NO2;

[0309] m is 0, 1, or 2; and

[0310] Z 1 and Z 2together form a moiety derived from an alpha-hydroxy carboxylic acid, wherein the atom attached to boron in each case is an oxygen atom; or Z 1 together form a moiety derived from a beta-hydroxy carboxylic acid, wherein the atom attached to boron in each case is an oxygen atom. 2 together form a moiety derived from a beta-hydroxy carboxylic acid, wherein the atom attached to boron in each case is an oxygen atom.

[0311] 88. The liquid pharmaceutical composition of item 87, wherein the tonicity adjusting agent, when present, is sodium chloride.

[0312] 89. The liquid pharmaceutical composition of item 87, wherein the buffering agent is sodium citrate and citric acid.

[0313] 90. The liquid pharmaceutical composition of item 87, wherein:

[0314] the alpha-hydroxy carboxylic acid or the beta-hydroxy carboxylic acid is citric acid;

[0315] A is 0;

[0316] R a is isobutyl;

[0317] R a1 is hydrogen, C 1-6 aliphatic, -(CH2) m -CH2-R B or -(CH2) m -CH(R 5a )-OR 5b ;

[0318] P is R c -C(O)-;

[0319] R c is -R D ;

[0320] m is 0 or 1;

[0321] the buffering agent is sodium citrate and citric acid; and

[0322] the tonicity adjusting agent, when present, is sodium chloride.

[0323] 91. A unit dose liquid pharmaceutical composition comprising a compound of formula (I-1):

[0324]

[0325] wherein:

[0326] the compound of formula (I-1) is present at a concentration of about 0.5 mg / ml to about 3 mg / ml of the compound of formula (VIII-1).

[0327] 92. The unit-dose liquid pharmaceutical composition according to clause 91, further comprising sodium citrate, citric acid, and sodium chloride.

[0328] 93. Use of a therapeutically effective amount of the pharmaceutical composition according to any one of clauses 24, 52, 57, 58, 59, 72, 77, and 87 in the manufacture of a medicament for the treatment of cancer.

[0329] In one aspect, the present application provides a compound of general formula (I):

[0330]

[0331] or a pharmaceutically acceptable salt thereof, wherein:

[0332] A is 0, 1, or 2;

[0333] P is hydrogen or an amino capping moiety;

[0334] R a is hydrogen, C 1-6 aliphatic, C 1-6 fluoroaliphatic, -(CH2) m -CH2-R B , -(CH2) m -CH2-NHC(=NR 4 )NH-Y, -(CH2) m -CH2-CON(R 4 )2, -(CH2) m -CH2-N(R 4 )CON(R 4 )2, -(CH2) m -CH(R 6 )N(R 4 )2, -(CH2) m -CH(R 5a )-OR 5b , or -(CH2) m -CH(R 5 )-SR 5 ;

[0335] R a1 is hydrogen, C 1-6 aliphatic, C 1-6 fluoroaliphatic, -(CH2) m -CH2-R B , -(CH2) m -CH2-NHC(=NR 4 )NH-Y, -(CH2) m -CH2-CON(R 42、-(CH2) m -CH2-N(R 4 CON(R) 4 2、-(CH2) m -CH(R 6 )N(R 4 2、-(CH2) m -CH(R 5a )-OR 5b Or -(CH2) m -CH(R 5 )-SR 5 ;

[0336] Each R a2 Independently hydrogen, C 1-6 aliphatic, C 1-6 Fluorine aliphatic, -(CH2) m -CH2-R B -(CH2) m -CH2-NHC(=NR 4 )NH-Y、-(CH2) m -CH2-CON(R 4 2、-(CH2) m -CH2-N(R 4 CON(R) 4 2、-(CH2) m -CH(R 6 )N(R 4 2、-(CH2) m -CH(R 5a )-OR 5b Or -(CH2) m -CH(R 5 )-SR 5 ;

[0337] Each R B Independent of substituted or unsubstituted monocyclic or bicyclic systems;

[0338] Each R 4 Independently hydrogen or substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclic groups; or two R groups on the same nitrogen atom. 4 The nitrogen atom is bonded together to form a substituted or unsubstituted 4- to 8-membered heterocyclic base ring, which, in addition to the nitrogen atom, has 0-2 cyclic heteroatoms independently selected from the group consisting of N, O and S;

[0339] Each R 5 It is independently hydrogen or a substituted or unsubstituted aliphatic, aryl, heteroaryl or heterocyclic group;

[0340] each R is independently hydrogen or substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl; 5a independently hydrogen or substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl;

[0341] each R is independently hydrogen or substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl; 5b independently hydrogen or substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl;

[0342] each R is independently hydrogen or substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl; 6 independently substituted or unsubstituted aliphatic, aryl, or heteroaryl;

[0343] Y is hydrogen, -CN, or -NO2;

[0344] m is 0, 1, or 2; and

[0345] Z 1 forms, together with Z 2 a moiety derived from an alpha-hydroxy carboxylic acid, wherein the atom attached to boron in each instance is an oxygen atom; or Z 1 forms, together with Z 2 a moiety derived from a beta-hydroxy carboxylic acid, wherein the atom attached to boron in each instance is an oxygen atom.

[0346] In another aspect, the present application provides a pharmaceutical composition comprising a compound of Formula (I) or a crystalline form thereof and other excipients described herein suitable for preparing an oral pharmaceutical dosage form.

[0347] In another aspect, the present application provides a pharmaceutical composition comprising a compound of Formula (I) or a crystalline form thereof and other excipients described herein suitable for preparing a lyophilized powder pharmaceutical dosage form.

[0348] In another aspect, the present application provides a pharmaceutical composition comprising a compound of Formula (I) or a crystalline form thereof and other excipients described herein suitable for preparing a liquid pharmaceutical dosage form.

[0349] In another aspect, the present application provides a pharmaceutical composition comprising a compound of Formula (I) or a crystalline form thereof, a filler, and optionally a lubricant.

[0350] In another aspect, the present application provides a pharmaceutical composition comprising a compound of Formula (I) or a crystalline form thereof, a filler, optionally a lubricant, optionally a glidant, and optionally a buffer.

[0351] In another aspect, the present application provides a pharmaceutical composition comprising a compound of Formula (I) or a crystalline form thereof, a bulking agent, and a buffer.

[0352] In another aspect, the present application provides a method of preparing a pharmaceutical composition of the present application.

[0353] In another aspect, the present application provides a method of treating a patient having a proteasome-mediated disorder or at risk of developing or experiencing a recurrence of a proteasome-mediated disorder using a pharmaceutical composition of the present application.

[0354] In another aspect, the present application provides a method of treating cancer using a pharmaceutical composition of the present application.

[0355] Definitions

[0356] Unless explicitly stated otherwise, the term "proteasome" is intended to refer to constitutive proteasomes as well as immunoproteasomes.

[0357] The term "aliphatic" or "aliphatic group", as used herein, means a straight-chain, branched- chain or cyclic hydrocarbon group that is completely saturated or that contains one or more units of 1-12 hydrocarbons that are completely saturated or that contain one or more units of unsaturation, but that are not aromatic. For example, suitable aliphatic groups include straight- chain, branched- chain or cyclic alkyl, alkenyl or alkynyl groups, and hybrids thereof such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl or (cycloalkyl)alkenyl groups. In various embodiments, aliphatic groups have 1 to 12, 1 to 8, 1 to 6, 1 to 4 or 1 to 3 carbon atoms.

[0358] The terms "alkyl", "alkenyl" and "alkynyl", used alone or as part of a larger moiety, refer to straight- chain or branched-chain aliphatic groups having 1 to 12 carbon atoms. For purposes of this application, the term "alkyl" is used when the carbon atom attaching the aliphatic group to the rest of the molecule is a saturated carbon atom. However, the alkyl group can include unsaturation at other carbon atoms. Thus, alkyl includes, but is not limited to, methyl, ethyl, propyl, allyl, propargyl, butyl, pentyl and hexyl groups.

[0359] For purposes of this application, the term "alkenyl" is used when the carbon atom attaching the aliphatic group to the rest of the molecule is part of a carbon-carbon double bond. Alkenyl groups include, but are not limited to, ethenyl, 1-propenyl, 1-butenyl, 1-pentenyl and 1-hexenyl.

[0360] For purposes of this application, the term "alkynyl" is used when the carbon atom attaching the aliphatic group to the rest of the molecule is part of a carbon-carbon triple bond. Alkynyl groups include, but are not limited to, ethynyl, 1-propynyl, 1-butynyl, 1-pentynyl and 1-hexynyl.

[0361] The term "alicyclic," used alone or as part of a larger moiety (e.g., "aralkyl," "aralkyloxy," or "aryloxyalkyl"), refers to a saturated or partially unsaturated cyclic aliphatic ring system having 3 to about 14 members, wherein the aliphatic ring system is optionally substituted. In certain embodiments, alicyclic is a monocyclic hydrocarbon having 3-8 or 3-6 ring carbon atoms. Non-limiting examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cycloheptenyl, cyclooctyl, cyclooctenyl, and cyclooctadienyl. In certain embodiments, alicyclic is a bridged or fused bicyclic hydrocarbon having 6-12, 6-10, or 6-8 ring carbon atoms, wherein any individual ring in the bicyclic system has 3-8 members.

[0362] In certain embodiments, two adjacent substituents on an alicyclic ring are taken together with the interjacent ring atoms to form an optionally substituted fused 5- to 6-membered aromatic ring or 3- to 8-membered non-aromatic ring having 0-3 ring heteroatoms selected from the group consisting of O, N, and S. Thus, the term "alicyclic" includes aliphatic rings fused to one or more aryl, heteroaryl, or heterocyclyl rings, wherein the linkage group or point of attachment is on the aliphatic ring. Non-limiting examples include indanyl, 5,6,7,8-tetrahydroquinoxalinyl, decahydronaphthalenyl, or tetrahydronaphthalenyl.

[0363] The terms "aryl" and "ar-", used alone or as part of a larger moiety (e.g., "aralkyl," "aralkyloxy," or "aryloxyalkyl"), refer to a C6to C 14 An aromatic hydrocarbon comprising one to three rings each optionally substituted. Preferably, aryl is a C 6-10 An aromatic hydrocarbon comprising one to three rings each optionally substituted. Preferably, aryl is a C The term "aryl," as used herein, includes (but is not limited to) phenyl, naphthyl, and anthryl. In certain embodiments, two adjacent substituents on an aryl ring are taken together with the interjacent ring atoms to form an optionally substituted fused 5- to 6-membered aromatic ring or 4- to 8-membered non-aromatic ring having 0-3 ring heteroatoms selected from the group consisting of O, N, and S. Thus, the term "aryl" as used herein includes groups in which an aryl ring is fused to one or more heteroaryl, alicyclic, or heterocyclyl rings, wherein the linkage group or point of attachment is on the aromatic ring. Non-limiting examples of such fused ring systems include indolyl, isoindolyl, benzothiophenyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzthiazolyl, quinolinyl, isoquinolinyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, fluorenyl, dihydroindenyl, phenanthridinyl, tetrahydronaphthalenyl, indolinyl, phenoxazinyl, benzodioxanyl, and benzodioxolyl. The aryl group can be monocyclic, bicyclic, tricyclic, or polycyclic, preferably monocyclic, bicyclic, or tricyclic, more preferably monocyclic or bicyclic. The term "aryl" is used interchangeably with the terms "aryl moiety" and "aryl ring."

[0364] "Aryalkyl" or "arylalkyl" includes an aryl group covalently attached to an alkyl group, either of which is independently optionally substituted. Preferably, the arylalkyl is a C 6-10 aryl(C 1-6 )alkyl, C 6-10 aryl(C 1-4 )alkyl, or C 6-10 aryl(C 1-3 )alkyl, including, but not limited to, benzyl, phenethyl, and naphthylmethyl.

[0365] The terms "heteroaryl" and "heteroar-," by themselves or as part of a larger moiety, e.g., heteroaralkyl, or "heteroaralkoxy," refer to groups having 5 to 14 ring atoms, preferably 5, 6, 9, or 10 ring atoms; having 6, 10, or 14 π electrons shared in a cyclic array; and having, in addition to carbon atoms, from one to four heteroatoms. The term "heteroatom" refers to nitrogen, oxygen, or sulfur, and includes any oxidized form of nitrogen or sulfur, and any quaternized form of a basic nitrogen. Thus, the term "nitrogen" when used in reference to a heteroaryl group includes an oxidized nitrogen, e.g., as in pyridine N-oxide. Certain of the nitrogen atoms of 5-membered heteroaryls are also substitutable, as further defined below. Heteroaryl includes, by way of example, groups derived from thiophene, furan, pyrrole, imidazole, pyrazole, triazole, tetrazole, oxazole, isoxazole, oxadiazole, thiazole, isothiazole, thiadiazole, pyridine, pyridazine, pyrimidine, pyrazine, indolizine, naphthylidine, pteridine, azaindole, benzofused pyrrol, imidazole, oxazole, thiazole, triazole, pyrrolopyrimidine, purine, and triazolopyrimidine. As used herein, the phrase "group derived from" means a monovalent group resulting from removal of a hydrogen group from a parent heteroaromatic ring system. The point of attachment of the group (i.e., the point of attachment of the heteroaryl group to the remainder of the molecule) can be made at any substitutable position on any ring of the parent heteroaryl ring system.

[0366] In certain embodiments, two adjacent substituents on a heteroaryl group are taken together with the interjacent ring atoms to form an optionally substituted fused 5- to 6-membered aromatic ring or 4- to 8-membered non-aromatic ring having 0-3 ring heteroatoms selected from the group consisting of O, N, and S. Thus, as used herein, the terms "heteroaryl" and "heteroar- " also include groups in which the heteroaromatic ring is fused together with one or more aryl, alicyclic, or heterocyclic rings wherein the rings share an adjacent pair of ring atoms. Non-limiting examples include indolyl, isoindolyl, benzothiophenyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzthiazolyl, benzoxazolyl, quinolinyl, isoquinolinyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and pyrido[2,3-b]-l,4-oxazin-3(4H)-one. The heteroaryl group can be monocyclic, bicyclic, tricyclic, or polycyclic, preferably monocyclic, bicyclic, or tricyclic, more preferably monocyclic or bicyclic. The term "heteroaryl" is used interchangeably with the term "heteroaryl ring," either term including an optionally substituted ring. The term "heteroaralkyl" refers to an alkyl group substituted with a heteroaryl group, wherein the alkyl and heteroaryl portions independently are optionally substituted.

[0367] As used herein, the terms "aromatic ring" and "aromatic ring system" refer to an optionally substituted monocyclic, bicyclic, or tricyclic ring group having from 0-6, preferably 0-4, ring heteroatoms and having 6, 10, or 14 π electrons shared in a cyclic array. Thus, the terms "aromatic ring" and "aromatic ring system" encompass both aryl and heteroaryl groups.

[0368] As used herein, the terms "heterocycle," "heterocyclyl," "heterocyclic group" are used interchangeably and refer to a stable 3- to 7-membered monocyclic or fused 7- to 10-membered or bridged 6- to 10-membered heterocyclic moiety, which is either saturated or partially unsaturated and which has, in addition to carbon atoms, one or more, preferably one to four, heteroatoms as set forth in the definition below. The term "nitrogen" when used in reference to a ring atom of a heterocyclic ring includes substituted nitrogen. For example, in a heterocyclyl ring having 1-3 heteroatoms selected from the group consisting of oxygen, sulfur, and nitrogen, the nitrogen can be N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl), or +NR (as in N-substituted pyrrolidinyl). The heterocycle can be attached at any heteroatom or carbon atom that results in a stable structure, and any of the ring atoms can be optionally substituted. Examples of such saturated or partially unsaturated heterocyclyl groups include, but are not limited to, tetrahydrofuranyl, tetrahydrothiophenyl, pyrrolidinyl, pyrrolidinonyl, piperidinyl, pyrrolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and quinuclidinyl.

[0369] In certain embodiments, two adjacent substituents on a heterocycle are taken together with the intervening ring atoms to form an optionally substituted fused 5- to 6-membered aromatic ring or 3- to 8-membered non-aromatic ring having 0-3 ring heteroatoms selected from the group consisting of O, N, and S. Thus, the terms "heterocycle," "heterocyclyl," "heterocyclyl ring," "heterocyclic group," "heterocyclic moiety" are used interchangeably herein and include groups in which a heterocyclyl ring is fused to one or more aryl, heteroaryl, or cycloaliphatic rings, such as indolinyl, 3H- indolyl, chromanonyl, phenanthridinyl, or tetrahydroquinolinyl, wherein the linkage is through a heterocyclyl ring atom or through a carbon atom of the heterocyclyl ring. The heterocyclyl group can be monocyclic, bicyclic, tricyclic, or polycyclic, preferably monocyclic, bicyclic, or tricyclic, more preferably monocyclic or bicyclic. The term "heterocyclylalkyl" refers to an alkyl group substituted with a heterocyclyl group, wherein the alkyl and heterocyclyl moieties are independently optionally substituted.

[0370] As used herein, the term "partially unsaturated" refers to a ring moiety that includes at least one double or triple bond between ring atoms. The term "partially unsaturated" is intended to encompass rings having multiple sites of unsaturation, but is not intended to include aryl or heteroaryl moieties as defined herein.

[0371] The terms "haloaliphatic," "haloalkyl," "haloalkenyl," and "haloalkoxy" mean an aliphatic, alkyl, alkenyl, or alkoxy group, respectively, optionally substituted with one or more halogen atoms. As used herein, the term "halogen" or "halo" means F, Cl, Br, or I. The term "fluoroaliphatic" refers to a haloaliphatic group in which the halogen is fluoro, including perfluorinated aliphatic groups. Examples of fluoroaliphatic groups include, but are not limited to, fluoromethyl, difluoromethyl, trifluoromethyl, 2-fluoroethyl, 2,2,2-trifluoroethyl, 1,1,2-trifluoroethyl, 1,2,2-trifluoroethyl, and pentafluoroethyl.

[0372] The term "linking group" or "linker" means an organic moiety that links two moieties of a compound. Linkers typically comprise an atom, e.g., oxygen or sulfur; a unit, e.g., -NH-, -CH2-, -C(O)-, -C(O)NH- or a chain of atoms, e.g., an alkylene chain. Linkers typically have a molecular mass in the range of about 14 to 200, preferably in the range of 14 to 96, and a length of up to about six atoms. In certain embodiments, the linker is a C 1-6 alkylene chain.

[0373] The term "alkylene" refers to a divalent alkyl group. An "alkylene chain" is a polymethylene, i.e., -(CH2) y wherein Y is a positive integer, preferably a positive integer of 1 to 6, 1 to 4, 1 to 3, 1 to 2, or 2 to 3. A substituted alkylene chain is a polymethylene in which one or more methylene hydrogen atoms are replaced by a substituent. Suitable substituents include those substituents described below for substituted aliphatic groups. An alkylene chain can also be substituted at one or more positions by an aliphatic or substituted aliphatic group.

[0374] A functional group can also optionally be inserted into an alkylene chain. When an internal methylene unit is replaced by a functional group, the functional group is "inserted" into the alkylene chain. Examples of suitable "insertion functional groups" include -C(R*)=C(R*)-, -C≡C-, -O-, -S-, -S(O)-, -S(O)2-, -S(O)2N(R + )-, -N(R*)-, -N(R + )CO-, -N(R + )C(O)N(R + )-, -N(R + )C(=NR + )-N(R + )-, -N(R + )-C(=NR + )-, -N(R + )CO2-, -N(R + )SO2-, -N(R + )SO2N(R + )-, -OC(O)-, -OC(O)O-, -OC(O)N(R + )-, -C(O)-, -CO2-, -C(O)N(R + )-, -C(O)-C(O)-, -C(=NR + )-N(R + )-, -C(NR + )=N-, -C(=NR + )-O-, -C(OR*)=N-, -C(R o) = N-O- or -N(R + ) = N-O- or -N(R + ) = N-O- or -N(R + independently hydrogen or optionally substituted aliphatic, aryl, heteroaryl, or heterocyclyl; or two R + on the same nitrogen atom are taken together to form a 5-8 membered aromatic or nonaromatic ring having, in addition to the nitrogen atom, 0-2 ring heteroatoms selected from the group consisting of N, O, and S. Each R*is independently hydrogen or optionally substituted aliphatic, aryl, heteroaryl, or heterocyclyl. Each R o independently optionally substituted aliphatic, aryl, or heteroaryl.

[0375] C 3-6 Examples of alkylene chains that are "interrupted" by -O- include -CH2OCH2-, -CH2O(CH2)2-, -CH2O(CH2)3-, -CH2O(CH2)4-, -(CH2)2OCH2-, -(CH2)2O(CH2)2-, -(CH2)2O(CH2)3-, -(CH2)3O(CH2)-, -(CH2)3O(CH2)2-, and -(CH2)4O(CH2)-. Other examples of alkylene chains that are "interrupted" by a functional group include -CH2Z*CH2-, -CH2Z*(CH2)2-, -CH2Z*(CH2)3-, -CH2Z*(CH2)4-, -(CH2)2Z*CH2-, -(CH2)2Z*(CH2)2-, -(CH2)2Z*(CH2)3-, -(CH2)3Z*(CH2)-, -(CH2)3Z*(CH2)2-, and -(CH2)4Z*(CH2)-, where Z*is one of the "interrupting" functional groups listed above.

[0376] Those of ordinary skill in the art will appreciate that certain combinations are not sufficiently stable for pharmaceutical use when an interrupted alkylene chain is attached to a functional group. Only stable or chemically feasible compounds are within the scope of the present application. A stable or chemically feasible compound is one that maintains its integrity long enough to be used for therapeutic or prophylactic administration to a patient. Preferably, the chemical structure does not change materially when kept at temperatures less than -70 °C, less than -50 °C, less than -20 °C, less than 0 °C, or less than 20 °C, in the absence of moisture or other chemically reactive conditions for at least one week.

[0377] As used herein, the term "substituted" means that a hydrogen base in the named moiety is replaced with a radical of a particular substituent, limited only by the valence of the atom. The term "may be substituted" when used in respect to a specified atom means that attached to the atom is a hydrogen radical, which can be replaced with a radical of a suitable substituent.

[0378] As used herein, the phrase "one or more substituents" means a number of substituents, equal to one up to the maximum number possible, based on the number of available bonding sites. Unless otherwise indicated, an optionally substituted group can have substituents at each substitutable position, and the substituents can be the same or different.

[0379] As used herein, the term "independently" or "independently selected" means that the same or different values can be selected for multiple instances of a given variable in a single compound.

[0380] An aryl group (including the aryl portion of aralkyl, aralkyloxy, aryloxyalkyl, and the like) or a heteroaryl group (including the heteroaryl portion of heteroaralkyl and heteroaralkyloxy, and the like) can contain one or more substituents. Examples of suitable substituents on an unsaturated carbon atom of an aryl or heteroaryl group include -halo, -NO2, -CN, -R*, -C(R*)=C(R*)2, -C≡C-R*, -OR*, -SR o , -S(O)R o , -SO2R o , -SO3R*, -SO2N(R + )2, -N(R + )2, -NR + C(O)R*, -NR + C(O)N(R + )2, -N(R + )C(=NR + )-N(R + )2, -N(R + )C(=NR + )-R o , -NR + CO2R o , -NR + SO2R o , -NR + SO2N(R + )2, -O-C(O)R*, -O-CO2R*, -OC(O)N(R + )2, -C(O)R*, -CO2R*, -C(O)-C(O)R*, -C(O)N(R + )2, -C(O)N(R + )-OR*, -C(O)N(R + )C(=NR + )-N(R + )2, -N(R + )C(=NR +)-N(R + -C(O)R*、-C(=NR) + )-N(R + )2、-C(=NR + -OR*, -N(R) + )-N(R + )2、-C(=NR + )-N(R + -OR*, -C(R) o )=N-OR*, -P(O)(R*)2, -P(O)(OR*)2, -OP(O)-OR* and -P(O)(NR + )-N(R + )2, where R o R + R* as defined above, or two adjacent substituents combined with their inserted atoms to form a 5-6 member unsaturated or partially unsaturated ring having 0-3 ring atoms selected from the group consisting of N, O and S.

[0381] The aliphatic group or non-aromatic heterocycle may be substituted with one or more substituents. Examples of suitable substituents on the saturated carbon of the aliphatic group or non-aromatic heterocycle include (but are not limited to) those substituents listed above for the unsaturated carbon of the aryl or heteroaryl group, and the following substituents: =O, =S, =C(R*)2, =NN(R*)2, =N-OR*, =N-NHC(O)R*, =N-NHCO2R o =N-NHSO2R o OR = NR*, where each R* and R o As defined above.

[0382] Suitable substituents on the substituted nitrogen atom of a heteroaryl or non-aromatic heterocycle include -R*, -N(R*)2, -C(O)R*, -CO2R*, -C(O)-C(O)R*-C(O)CH2C(O)R*, -SO2R*, -SO2N(R*)2, -C(=S)N(R*)2, -C(=NH)-N(R*)2, and -NR*SO2R*; wherein each R* is as defined above. The cyclic nitrogen atom of the heteroaryl or non-aromatic heterocycle can also be oxidized to form the corresponding N-hydroxyl or N-oxide compound. A non-limiting example of said heteroaryl having an oxidized cyclic nitrogen atom is an N-oxo-pyridyl group.

[0383] The term "about" is used herein to mean approximately, roughly, roughly, or approximately. When the term "about" is used in conjunction with a numerical range, it modifies that range by extending the limits above and below the listed value. Generally, the term "about" is used herein to modify a numerical value so that its deviation above and below the stated value is 10%.

[0384] As used herein, the term "comprising" means "including, but not limited to."

[0385] It will be apparent to those skilled in the art that certain compounds of the present application can exist in tautomeric forms. All such tautomeric forms are within the scope of the present application. Unless otherwise stated, structures depicted herein are also meant to include all geometric (or conformational) isomers, for example (Z) and (E) double bond isomers, and (Z) and (E) conformational isomers; as well as the

[0386] Unless otherwise stated, structures depicted herein are also meant to include all isomeric forms of the structure, e.g., tautomers and geometric (or conformational isomers). Therefore, structural formulas recited herein are intended to represent only one form present and all possible individual isomers and mixture thereof. 13 C-enriched carbon or 14 C-enriched carbon for carbon atoms are within the scope of this application.

[0387] As used herein, the term "seeding" is used to refer to the addition of crystalline material to initiate crystallization or recrystallization.

[0388] When compounds crystallize from solution or slurry, they can crystallize in different spatial lattice arrangements, a property known as "polymorphism." Each crystalline form is a "polymorph." While polymorphs of a given substance have the same chemical composition, they can differ from each other in one or more physical properties, such as solubility and dissolution rate, true density, melting point, crystal shape, compaction behavior, flow properties, and / or solid state stability.

[0389] As used herein, the term "solvate" or "solvation" means the physical association of one or more solvent molecules with a compound. This physical association includes hydrogen bonding. In certain instances, the solvent molecules are incorporated in the crystalline lattice of the compound. "Solvate" or "solvation" encompasses both solution-phase and isolatable solvates. Representative solvates include, for example, hydrates, ethanolates, or methanolates. The physical properties of one solvate will typically differ from those of another solvate, as well as from those of the non-solvated form of the compound. Because the chemical composition also differs between solvates, these forms are referred to as "pseudo-polymorphs."

[0390] As used herein, the term "hydrate" is a solvate of H2O in which the solvent molecule is present in a definite stoichiometric amount and can, for example, include a hemihydrate, monohydrate, dihydrate, or trihydrate. As used herein, the term "anhydrate" is a compound of the application that does not contain H2O incorporated into its crystal lattice.

[0391] As used herein, "crystalline" refers to a solid having a highly regular chemical structure. In particular, a crystalline compound can be produced in one or more single crystalline forms of the compound. For the purposes of the present application, the terms "single crystalline form" or "crystalline form" are used interchangeably and distinguish between crystals having different properties (e.g., different XRPD patterns, different DSC scan results). Thus, each unique polymorph and pseudopolymorph of a compound is considered a unique single crystalline form herein.

[0392] "Substantially crystalline" refers to a compound that can be at least a particular weight percent crystalline. The particular weight percent is 10%, 20%, 30% 40%, 50%, 60%, 70%, 75%, 80%, 85%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%, or any percent between 10% and 100%. In certain embodiments, substantially crystalline refers to a compound that is at least 70% crystalline. In other embodiments, substantially crystalline refers to a compound that is at least 90% crystalline.

[0393] "Substantially pure" refers to a compound that can be at least a particular weight percent of the compound. The particular weight percent is about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or about 99.5%.

[0394] Unless explicitly stated otherwise, structures depicted herein are intended to include all atropisomers and all stereoisomeric forms of the structures.

[0395] As used herein, the terms "Compound (I-1)" and "4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetylamino)-3-methylbutyl)-6-oxo-1,3,2-dioxaborane-4-carboxylic acid" are used interchangeably and include all crystalline forms. Both terms refer to the compound produced in Example 1 and Example 1A of the Examples below, including both Form 1 and Form 2.

[0396] As used herein, the term "Compound (I-1) Form 2" is used interchangeably with "4-(R,S)-(carboxymethyl)-2-((R)-l-(2-(2,5-dichlorobenzamido)acetylamino)-3- methylbutyl)-6-oxo-l,3,2-dioxaborinane-4-carboxylic acid Form 2." Both terms refer to the crystalline Form 2 prepared in Example 1 of the Examples and in Example 1A.

[0397] As used herein, the term "Compound of Formula (VIII-1)" is used interchangeably with "(R)-l-((2,5-dichlorobenzamido)acetylamino)-3-methylbutylboronic acid." The compound of Formula (VIII-1) is disclosed in U.S. Patent No. 7,442,830 and WO 09 / 020448.

[0398] As used herein, the term "Compound of Formula (I-15)," "Compound (I-15)," and "(I-15)" are used interchangeably and refer to the citric acid ester of Compound (VIII-15), and the compound produced in Example 15 of the Examples.

[0399] As used herein, the term "anhydride" with respect to boronic acids (e.g., compounds of Formula (VIII)) refers to a compound formed by the combination of two or more molecules of a boronic acid compound with the loss of one or more water molecules. When mixed with water, the boronic anhydride compound hydrates to release free boronic acid compound. In various embodiments, boronic anhydrides can comprise two, three, four, or more boronic acid units, and can have a cyclic or linear configuration. Non-limiting examples of oligomeric boronic anhydrides of the peptide boronic acid compounds of the present invention are illustrated below:

[0400]

[0401] In Formulas (1) and (2), the variable nn is an integer from 0 to about 10, preferably 0, 1, 2, 3, or 4. In certain embodiments, the boronic anhydride compound comprises a cyclic trimer of Formula (2) ("cycloboroxane") wherein nn is 1. The variable W has the formula (3):

[0402]

[0403] wherein P, R a2 , A, R a1 and R a are as defined herein.

[0404] As used herein, the total weight of a single oral pharmaceutical dosage form is determined by adding together the weight of all of the components of the oral pharmaceutical dosage form and does not include the weight of any coating that can optionally be applied to the oral pharmaceutical dosage form after it is formed. The total weight of a single oral pharmaceutical dosage form is used as the basis for calculating the weight percentage of each component that makes up the oral pharmaceutical dosage form.

[0405] As used herein, "low moisture" as used with respect to an excipient, such as a filler, means that the excipient has a water content of about 0.5% to about 4%. The term "low moisture" can be used interchangeably with the term "low water."

[0406] As used herein, the term "lyophilized powder," "cake," or "lyophilized cake" means any solid material obtained by lyophilizing an aqueous mixture.

[0407] As used herein, the term "tonicity adjusting agent" means an agent that helps to adjust the osmotic pressure of a liquid or solution.

[0408] As used herein, the term "boronate ester" is used interchangeably with "boronic ester" and means a compound containing a -B(Z 1 )(Z 2 ) moiety, wherein Z 1 and Z 2 together form a moiety wherein the atom attached to the boron in each instance is an oxygen atom.

[0409] In certain embodiments, the boronate moiety is a 5-membered ring. In certain other embodiments, the boronate moiety is a 6-membered ring. In certain other embodiments, the boronate moiety is a mixture of 5-membered rings and 6-membered rings.

[0410] As used herein, the term "a-hydroxy carboxylic acid" means a compound containing a hydroxyl group directly attached to a carbon atom that is in the a position relative to the carboxylic acid group. As used herein, the term "a-hydroxy carboxylic acid" is not intended to be limited to compounds having only one hydroxyl group and one carboxylic acid group.

[0411] As used herein, the term "β-hydroxy carboxylic acid" means a compound containing a hydroxyl group directly attached to a carbon atom that is in the β position relative to the carboxylic acid group. As used herein, the term "β-hydroxy carboxylic acid" is not intended to be limited to compounds having only one hydroxyl group and one carboxylic acid group.

[0412] As used herein, the term "moiety derived from an α-hydroxycarboxylic acid" refers to a moiety formed by removal of a hydrogen atom from the carboxylic acid within an α-hydroxycarboxylic acid and removal of a hydrogen atom from the hydroxyl group directly attached to the carbon atom located in the α position relative to the carboxylic acid group. As used herein, the term "moiety derived from a β-hydroxycarboxylic acid" refers to a moiety formed by removal of a hydrogen atom from the carboxylic acid within a β-hydroxycarboxylic acid and removal of a hydrogen atom from the hydroxyl group directly attached to the carbon atom located in the β position relative to the carboxylic acid group. BRIEF DESCRIPTION OF DRAWINGS

[0413] Figure 1 Powder X-ray diffraction pattern of 4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5- dichlorobenzamido)acetylamino)-3-methylbutyl)-6-oxo-1,3,2-dioxaborolan-4- carboxylic acid (1-1) Form 1;

[0414] Figure 2 Differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) plot of 4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetylamino)-3- methylbutyl)-6-oxo-1,3,2-dioxaborolan-4-carboxylic acid (1-1) Form 1;

[0415] Figure 3 Powder X-ray diffraction pattern of 4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5- dichlorobenzamido)acetylamino)-3-methylbutyl)-6-oxo-1,3,2-dioxaborolan-4- carboxylic acid (1-1) Form 2;

[0416] Figure 4 Differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) plot of 4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetylamino)-3- methylbutyl)-6-oxo-1,3,2-dioxaborolan-4-carboxylic acid (1-1) Form 2;

[0417] Figure 5 Powder X-ray diffraction pattern of 2,5-dichloro-N-[2-({(1R)-3-methyl-1-[(4S)-4- methyl-5-oxo-1,3,2-dioxaborolan-2-yl]butyl}amino)-2-oxoethyl]benzamide (1-7);

[0418] Figure 6a powder X-ray diffraction pattern of 2,5-dichloro-N-(2-{[(1 R)-3-methyl-1 -(4-oxo- 4H-1,3,2-benzodioxaborol-2-yl)butyl]amino}-2-oxoethyl)benzamide (1-13);

[0419] Figure 7 a powder X-ray diffraction pattern of 4-(R,S)-(carboxymethyl)-2-((R)-1 -(2-(2,5- dichlorobenzamido)acetamido)-3-methylbutyl)-6-oxo-1,3,2-dioxaborolan-4- carboxylic acid (1-1 ) Form 2; and

[0420] Figure 8 a differential scanning calorimetry (DSC) plot of 4-(R,S)-(carboxymethyl)-2-((R)-1 -(2-(2,5- dichlorobenzamido)acetamido)-3-methylbutyl)-6-oxo-1,3,2-dioxaborolan-4- carboxylic acid (1-1 ) Form 2. DETAILED DESCRIPTION

[0421] In certain embodiments, the a-hydroxy acid is characterized by Formula (V):

[0422]

[0423] wherein R b3 and each R b4 is independently hydrogen, -CO2H, or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl.

[0424] In certain embodiments, R b3 and each R b4 is independently hydrogen, C 1-6 aliphatic, or -(CH2) p -CO2H, and p is 0, 1, or 2. In certain embodiments, R b3 and each R b4 is independently hydrogen or C 1-6 aliphatic. In certain such embodiments, R b3 and each R b4 is independently selected from the group consisting of hydrogen, methyl, ethyl, isopropyl, isobutyl, t-butyl, and cyclohexyl. In certain other embodiments, R b3 and each R b4 is independently hydrogen or -(CH2) p -CO2H. In certain such embodiments, p is 1. In certain other embodiments, R b3 and each R b4 is independently -(CH2) p-CO2H. In certain such embodiments, p is 1.

[0425] In certain embodiments, the a-hydroxycarboxylic acid is selected from the group consisting of glycolic acid, malic acid, hexahydromandelic acid, citric acid, 2-hydroxyisobutyric acid, mandelic acid, lactic acid, 2-hydroxy-3,3-dimethylbutyric acid, 2-hydroxy-3-methylbutyric acid, 2-hydroxyisocaproic acid, and diphenyl glycolic acid. In certain other embodiments, the a-hydroxycarboxylic acid is selected from the group consisting of glycolic acid, malic acid, hexahydromandelic acid, citric acid, 2-hydroxyisobutyric acid, mandelic acid, lactic acid, 2-hydroxy-3,3-dimethylbutyric acid, 2-hydroxy-3-methylbutyric acid, 2-hydroxyisocaproic acid, tartaric acid, and diphenyl glycolic acid. In certain embodiments, the a-hydroxycarboxylic acid is citric acid. Certain other non-limiting examples of a-hydroxycarboxylic acids include glucoheptonic acid, gluconic acid, lactobionic acid, and galactaric acid.

[0426] In certain embodiments, the β-hydroxy acid is characterized by Formula (VI):

[0427]

[0428] wherein R b1 and R b2 each independently is hydrogen, -CO2H, -OH, or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl; R b3 and R b4 each independently is hydrogen, -CO2H, or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl;

[0429] or R b2 and R b4 each independently is hydrogen, and R b1 and R b3 taken together with the carbon atom to which they are attached form an unsubstituted or substituted 4- to 8-membered non-aromatic ring having 0-3 ring heteroatoms selected from the group consisting of O, N, and S, wherein the ring can optionally be fused to an unsubstituted or substituted 4- to 8-membered non-aromatic ring or 5- to 6-membered aromatic ring having 0-3 ring heteroatoms selected from the group consisting of O, N, and S;

[0430] or R b2 and R b4 are absent, and R b1 and R b3together with the carbon atom to which they are attached form an unsubstituted or substituted 5- to 6-membered aromatic ring having 0-3 ring heteroatoms selected from the group consisting of O, N, and S, wherein the ring can be optionally fused to an unsubstituted or substituted 4- to 8-membered non-aromatic ring or a 5- to 6-membered aromatic ring having 0-3 ring heteroatoms selected from the group consisting of O, N, and S.

[0431] In certain embodiments, R b1 and R b2 each independently is hydrogen, C 1-6 aliphatic, -(CH2) p -OH, or -(CH2) p -CO2H, and p is 0, 1, or 2. In certain such embodiments, R b1 and R b2 each is hydrogen. In certain other such embodiments, R b1 is -OH and R b2 is hydrogen.

[0432] In certain embodiments, R b3 and R b4 each independently is hydrogen, C 1-6 aliphatic, or -(CH2) p -CO2H, and p is 0, 1, or 2. In certain embodiments, R b3 and R b4 each independently is hydrogen or C 1-6 aliphatic. In certain such embodiments, R b3 and R b4 each independently is selected from the group consisting of hydrogen, methyl, ethyl, isopropyl, isobutyl, t-butyl, and cyclohexyl. In certain other embodiments, R b3 and R b4 each independently is -(CH2) p -CO2H, and p is 0 or 1.

[0433] The variable p is 0, 1, or 2. In certain embodiments, p is 0 or 1. In certain embodiments, p is 0. In other certain embodiments, p is 1.

[0434] In certain embodiments, R b2 and R b4 are absent and R b1 and R b3 together with the carbon atom to which they are attached form a substituted or unsubstituted phenyl ring.

[0435] In certain embodiments, the β-hydroxycarboxylic acid is selected from the group consisting of malic acid, citric acid, 3-hydroxybutyric acid, β-hydroxyisovaleric acid, and salicylic acid. In certain other embodiments, the β-hydroxycarboxylic acid is selected from the group consisting of malic acid, citric acid, 3-hydroxybutyric acid, β-hydroxyisovaleric acid, tartaric acid, and salicylic acid. In certain embodiments, the β-hydroxycarboxylic acid is citric acid. Certain other non-limiting examples of β-hydroxycarboxylic acids include glucoheptonic acid, gluconic acid, lactobionic acid, and galactaric acid. Certain other non-limiting examples of β-hydroxycarboxylic acids include embonic acid, 1-hydroxy-2-naphthoic acid, and 3-hydroxy-2-naphthoic acid.

[0436] In certain embodiments, the α-hydroxy acid or β-hydroxy acid is selected from the group consisting of glycolic acid, malic acid, hexahydromandelic acid, 2-hydroxyisobutyric acid, citric acid, mandelic acid, lactic acid, 3-hydroxybutyric acid, β-hydroxyisovaleric acid, 2-hydroxy-3,3-dimethylbutyric acid, 2-hydroxy-3-methylbutyric acid, 2-hydroxyisocaproic acid, tartaric acid, salicylic acid, and benzolactic acid.

[0437] In certain embodiments, the compound of general formula (I) is characterized by formula (II):

[0438]

[0439] wherein:

[0440] the variables P, A, R a , R a1 , R a2 and n have the values described below and the variable R b1 , R b2 , R b3 and R b4 have the values described above.

[0441] In certain embodiments, any one of R b1 , R b2 , R b3 and R b4 may contain a functional group that can form another bond with a boron atom. In certain embodiments, the functional group is a carboxylic acid. In other certain embodiments, the functional group is a hydroxyl group.

[0442] In certain embodiments where the α-hydroxycarboxylic acid or β-hydroxycarboxylic acid is citric acid, the compound of general formula (I) is characterized by formula (III) or (IV) or mixtures thereof:

[0443]

[0444] wherein the variables P, A, R a , R a1 and R a2having the following values.

[0445] In certain other embodiments where the a-hydroxycarboxylic acid or the β- hydroxycarboxylic acid is citric acid, another bond can form between the carboxylic acid and the boron atom in Formula (III) or (IV). Without being bound by any theory of chemical bonding, in such embodiments, the compound of Formula (I) can be represented by Formula (Ilia) or (IVa), or mixtures thereof:

[0446]

[0447] wherein the variables P, A, R a , R a1 , and R a2 have the following values.

[0448] It will be appreciated that, without being bound by any theory of chemical bonding, other representations exist that can be used to describe this further bonding of the carboxylic acid and the boron atom in Formula (Ilia) and (IVa).

[0449] The following values describe the variables in any of Formula (I), (II), (III), (Ilia), (IV), or (IVa).

[0450] The variable P is hydrogen or an amino capping moiety. Non-limiting examples of amino capping moieties can be found in P.G.M. Wuts and T.W. Greene, Greene's Protective Groups in Organic Synthesis (4th Ed.), John Wiley & Sons, NJ (2007), and include, for example, acyl, sulfonyl, oxyacyl, and aminoacyl groups.

[0451] In certain embodiments, P is R c -C(O)-, R c -O-C(O)-, R c -N(R 4c )-C(O)-, R c -S(O)2- or R c -N(R 4c )-S(O)2-, where R c is selected from the group consisting of C 1-6 aliphatic, C 1-6 fluoroaliphatic, -R D , -T 1 -R D , and -T 1 -R 2c , and the variable T 1 , RD , R 2c , and R 4c have the following values.

[0452] The variable R 4c is hydrogen, C 1-4 alkyl, C 1-4 fluoroalkyl, or C 6-10 ar(C 1-4 alkyl), wherein the aryl moiety is substituted or unsubstituted. In certain embodiments, R 4c is hydrogen or C 1-4 alkyl. In certain embodiments, R 4c is hydrogen.

[0453] The variable T 1 is a C 3a alkylene chain substituted with 0-2 independently selected R 3b or R 1-6 , wherein the alkylene chain is optionally interrupted with -C(R 5 )=C(R 5 )-, -CºC-, or -O-. Each R 3a is independently selected from the group consisting of -F, -OH, -O(C 1-4 alkyl), -CN, -N(R 4 )2, -C(O)(C 1-4 alkyl), -CO2H, -CO2(C 1-4 alkyl), -C(O)NH2, and -C(O)-NH(C 1-4 alkyl). Each R 3b is independently C 3a aliphatic optionally substituted with R 7 or R 1-3 ; or two substituents R 3b on the same carbon atom are taken together to form a 3- to 6-membered alicyclic ring. Each R 7 is a substituted or unsubstituted aromatic group. In certain embodiments, T 1 is a C 1-4 alkylene chain.

[0454] The variable R 2c is halo, -OR 5 , -SR 6 , -S(O)R 6 , -SO2R 6 , -SO2N(R 4 )2, -N(R 4 )2, -NR 4 C(O)R 5 , -NR 4 C(O)N(R4 )2, -NR 4 CO2R 6 , -N(R 4 )SO2R 6 , -N(R 4 )SO2N(R 4 )2, -O-C(O)R 5 , -OC(O)N(R 4 )2, -C(O)R 5 , -CO2R 5 , or -C(O)N(R 4 )2, wherein:

[0455] each R 4 is independently hydrogen or optionally substituted aliphatic, aryl, heteroaryl, or heterocyclyl; or two R 4 on the same nitrogen atom are taken together to form an optionally substituted 4- to 8-membered heterocyclyl ring having, in addition to the nitrogen atom, 0-2 ring heteroatoms independently selected from the group consisting of N, O, and S;

[0456] each R 5 is independently hydrogen or optionally substituted aliphatic, aryl, heteroaryl, or heterocyclyl; and

[0457] each R 6 is independently optionally substituted aliphatic, aryl, or heteroaryl.

[0458] the variable R D is a substituted or unsubstituted aromatic, heterocyclyl, or alicyclic ring, any of which is optionally fused to a substituted or unsubstituted aromatic, heterocyclyl, or alicyclic ring. In certain embodiments, R D is substituted on the ring carbon atoms with 0-2 R d and 0-2 R 8d , and each substitutable ring nitrogen atom in R D is unsubstituted or substituted with -C(O)R 5 , -C(O)N(R 4 )2, -CO2R 6 , -SO2R 6 , -SO2N(R 4 )2, C 1-4 aliphatic, substituted or unsubstituted C 6-10 aryl, or C 6-10 ar(C 1-4 )alkyl, wherein the aryl moiety is substituted or unsubstituted. The variables R 4 , R 5 , and R 6 have the values described above. Each Rd independently selected from the group consisting of C 1-6 aliphatic, C 1-6 fluoroaliphatic, halo, -R 1d , -R 2d , -T 2 -R 1d and -T 2 -R 2d wherein the variables T 2 , R 1d , R 2d and R 8d have the values described below. In certain embodiments, each R d is independently selected from the group consisting of C 1-6 aliphatic, C 1-6 fluoroaliphatic and halo.

[0459] T 2 is a C 3a alkylene chain substituted with 0-2 independently selected R 3b or R 1-6 wherein the alkylene chain is optionally interrupted with -C(R 5 )=C(R 5 )-, -C≡C- or -O-. The variables R 3a , R 3b and R 5 have the values described above.

[0460] each R 1d is independently a substituted or unsubstituted aryl, heteroaryl, heterocyclyl or aliphatic ring.

[0461] each R 2d is independently -NO2, -CN, -C(R 5 )=C(R 5 )2, -C≡C-R 5 , -OR 5 , -SR 6 , -S(O)R 6 , -SO2R 6 , -SO2N(R 4 )2, -N(R 4 )2, -NR 4 C(O)R 5 , -NR 4 C(O)N(R 4 )2, -N(R 4 )C(=NR 4 )-N(R 4 )2, -N(R 4 )C(=NR 4 )-R 6-NR 4 CO2R 6 , -N(R 4 )SO2R 6 , -N(R 4 )SO2N(R 4 )2, -O-C(O)R 5 , -OC(O)N(R 4 )2, -C(O)R 5 , -CO2R 5 , -C(O)N(R 4 )2, -C(O)N(R 4 )-OR 5 , -C(O)N(R 4 )C(=NR 4 )-N(R 4 )2, -N(R 4 )C(=NR 4 )-N(R 4 )-C(O)R 5 , or -C(=NR 4 )-N(R 4 )2. The variables R 4 , R 5 , and R 6 have the values described above.

[0462] Each R 8d is independently selected from the group consisting of C 1-4 aliphatic, C 1-4 fluoroaliphatic, halo, -OH, -O(C 1-4 aliphatic), -NH2, -NH(C 1-4 aliphatic), and -N(C 1-4 aliphatic)2. In certain embodiments, each R 8d is independently C 1-4 aliphatic, C 1-4 fluoroaliphatic, or halo.

[0463] In certain embodiments, R D is a substituted or unsubstituted monocyclic or bicyclic ring system. In certain embodiments, R DThis refers to substituted or unsubstituted monocyclic or bicyclic systems selected from the group consisting of the following groups: furanyl, thiopheneyl, pyrroleyl, isoxazolyl, oxazolyl, thiazolyl, isothiazolyl, imidazolyl, pyrazolyl, oxadiazolyl, thiadiazolyl, phenyl, pyridinyl, pyridazinyl, pyrimidinyl, benzofuranyl, benzothiopheneyl, indolyl, benzooxazolyl, benzoisooxazolyl, benzoimidazolyl, indazoleyl, purinyl, naphthyl, quinolinyl, isoquinolinyl, cenolinyl, quinazolinyl, quinoxolinyl, phthalazinyl, naphthidyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, tetrahydroquinoxolinyl, and dihydrobenzoxazinyl. In some embodiments, R D The substituted or unsubstituted monocyclic or bicyclic system is selected from the group consisting of the following groups: phenyl, pyridyl, pyrimidinyl, pyrazinyl, naphthyl, benzimidazolyl, quinolinyl, isoquinolinyl, quinoxalinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, tetrahydroquinoxalinyl, and dihydrobenzoxazinyl.

[0464] In some embodiments, R D The substituted ring carbon atom in the middle is subjected to 0-1 R on the substituted carbon atom. d and 0-2 R 8d Replace; of which:

[0465] Each R d Independently for C 1-6 aliphatic, C 1-6 Fluorine aliphatic or halogenated groups; and

[0466] Each R 8d Independently for C 1-4 aliphatic, C 1-4 Fluorine aliphatic or halogenated.

[0467] In some embodiments, R D The substituted cyclic carbon atoms in the 0-1 R d and 0-2 R 8d Replace, of which:

[0468] T 1 For those that have not been replaced or have been R 3a Or R 3b Replacement C 1-3 Alkylene chain;

[0469] Each R 1d Independently, it is a substituted or unsubstituted aryl, heteroaryl, heterocyclic, or alicyclic ring; and

[0470] Each R 2d Independently for -OR 5 -SR 6 -S(O)R 6 -SO2R 6, -SO2N(R 4 )2, -N(R 4 )2, -NR 4 C(O)R 5 , -NR 4 C(O)N(R 4 )2, -O-C(O)R 5 , -OC(O)N(R 4 )2, -C(O)R 5 , -CO2R 5 , or -C(O)N(R 4 )2. The variables R 4 , R 5 and R 6 have the values described above.

[0471] In certain embodiments, the variable R d has the formula -Q-R E , wherein Q is -O-, -NH-, or -CH2-, and R E is a substituted or unsubstituted aryl, heteroaryl, heterocyclyl, or alicyclyl ring. In certain embodiments, R E is a substituted or unsubstituted phenyl, pyridyl, pyrimidinyl, pyrazinyl, piperidinyl, piperazinyl, or morpholinyl ring.

[0472] In certain embodiments, P has the formula R c -C(O)-, wherein R c is a C 1-4 alkyl, C 1-4 fluoroalkyl, or C 6-10 ar(C 1-4 )alkyl, wherein the aryl moiety is substituted or unsubstituted. In certain such embodiments, P is selected from the group consisting of acetyl, trifluoroacetyl, and phenylacetyl.

[0473] In certain other embodiments, P has the formula R D -C(O)-, wherein R D is a substituted or unsubstituted phenyl, pyridyl, pyrazinyl, pyrimidinyl, quinolinyl, or quinoxalinyl. In certain other embodiments, P has the formula R D -C(O), wherein R D is a phenyl, pyridyl, pyrazinyl, pyrimidinyl, naphthyl, quinolinyl, quinoxalinyl, benzimidazolyl, or dihydrobenzoxazinyl, substituted with 0-1 R d and 0-2 R 8d .

[0474] In certain embodiments, P has the formula R D -C(O)-, wherein R Dis 2-pyrazinyl. In other certain embodiments, P has the formula R D -C(O)-, wherein R D is 2,5-dichlorophenyl. In other certain embodiments, P has the formula R D -C(O)-, wherein R D is 6-phenyl-2-pyridinyl.

[0475] In certain other embodiments, P has the formula R c -SO2-, wherein R c is -R D or -T 1 -R D , wherein T 1 is C 1-4 alkylene and R D is phenyl, pyridinyl, pyrazinyl, pyrimidinyl, naphthyl, quinolinyl, quinoxalinyl, benzimidazolyl, or dihydrobenzoxazinyl substituted with 0-1 R d and 0-2 R 8d .

[0476] The variable R a is hydrogen, C 1-6 aliphatic, C 1-6 fluoroaliphatic, -(CH2) m -CH2-R B , -(CH2) m -CH2-NHC(=NR 4 )NH-Y, -(CH2) m -CH2-CON(R 4 )2, -(CH2) m -CH2-N(R 4 )CON(R 4 )2, -(CH2) m -CH(R 6 )N(R 4 )2, -(CH2) m -CH(R 5a )-OR 5b , or -(CH2) m -CH(R 5 )-SR 5 , wherein the variables R 4 , R 5 , and R 6 have the values described above and the variables R 5a , R 5b , R B , Y, and m have the values described below.

[0477] In certain embodiments, R a is hydrogen, C1-6 aliphatic, C 1-6 fluoroaliphatic, or -(CH2) m -CH2-R B In certain other embodiments, R a is C 1-6 aliphatic, or -(CH2) m -CH2-R B In certain other embodiments, R a is C 1-6 aliphatic. In other embodiments, R a is isobutyl, 1-naphthylmethyl, 2-naphthylmethyl, benzyl, 4-fluorobenzyl, 4-hydroxybenzyl, 4-(benzyloxy)benzyl, benzyl naphthylmethyl, or phenethyl. In certain embodiments, R a is isobutyl.

[0478] The variable R a1 is hydrogen, C 1-6 aliphatic, C 1-6 fluoroaliphatic, -(CH2) m -CH2-R B , -(CH2) m -CH2-NHC(=NR 4 )NH-Y, -(CH2) m -CH2-CON(R 4 )2, -(CH2) m -CH2-N(R 4 )CON(R 4 )2, -(CH2) m -CH(R 6 )N(R 4 )2, -(CH2) m -CH(R 5a )-OR 5b , or -(CH2) m -CH(R 5 )-SR 5 wherein the variables R 4 , R 5 and R 6 have the values described above and the variables R 5a , R 5b , R B , Y and m have the values described below.

[0479] In certain embodiments, R a1 is hydrogen, C 1-6 aliphatic, C 1-6 fluoroaliphatic, -(CH2) m -CH2-R B , or -(CH2)m -CH(R 5a )-OR 5b In certain other embodiments, R a1 is hydrogen, -(CH2) m -CH2-R B , or -(CH2) m -CH(R 5a )-OR 5b In certain other embodiments, R a1 is isobutyl, 1-naphthylmethyl, 2-naphthylmethyl, benzyl, 4-fluorobenzyl, 4-hydroxybenzyl, 4-(benzyloxy)benzyl, benzyl naphthylmethyl, or phenethyl.

[0480] In certain embodiments, R a1 is -CH2-R B In certain other embodiments, R a1 is -CH(R 5a )-OR 5b In other certain embodiments, R a1 is hydrogen.

[0481] The variable R a2 is hydrogen, C 1-6 aliphatic, C 1-6 fluoroaliphatic, -(CH2) m -CH2-R B , -(CH2) m -CH2-NHC(=NR 4 )NH-Y, -(CH2) m -CH2-CON(R 4 )2, -(CH2) m -CH2-N(R 4 )CON(R 4 )2, -(CH2) m -CH(R 6 )N(R 4 )2, -(CH2) m -CH(R 5a )-OR 5b , or -(CH2) m -CH(R 5 )-SR 5 wherein the variables R 4 , R 5 , and R 6 have the values described above and the variables R B , R 5a , R 5b , Y, and m have the values described below.

[0482] In certain embodiments, Ra2 hydrogen, C 1-6 aliphatic, C 1-6 fluoroaliphatic, -(CH2) m -CH2-R B or -(CH2) m -CH(R 5a )-OR 5b In certain other embodiments, R a2 is isobutyl, 1-naphtylmethyl, 2-naphtylmethyl, benzyl, 4-fluorobenzyl, 4-hydroxybenzyl, 4-(benzyloxy)benzyl, benzyl naphthylmethyl, or phenethyl.

[0483] each R B is independently a substituted or unsubstituted monocyclic or bicyclic ring system. In certain embodiments, each R B is independently a substituted or unsubstituted phenyl, pyridyl, indolyl, benzimidazolyl, naphthyl, quinolyl, quinoxalyl, or isoquinolyl ring. In certain embodiments, R B is a substituted or unsubstituted phenyl ring.

[0484] The variable Y is hydrogen, -CN, or -NO2. In certain embodiments, Y is -NO2.

[0485] The variable R 5a is hydrogen or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl. In certain embodiments, R 5a is hydrogen or a substituted or unsubstituted aliphatic group. In certain other embodiments, R 5a is hydrogen or C 1-6 aliphatic. In such embodiments, R 5a is selected from the group consisting of hydrogen, methyl, ethyl, isopropyl, and isobutyl. In certain such embodiments, R 5a is methyl.

[0486] The variable R 5b is hydrogen or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl. In certain embodiments, R 5b is hydrogen or a substituted or unsubstituted aliphatic group. In certain other embodiments, R 5b is hydrogen or C 1-6 aliphatic. In such embodiments, R 5b is selected from the group consisting of hydrogen, methyl, ethyl, isopropyl, and isobutyl. In certain such embodiments, R 5b is hydrogen.

[0487] The variable m is 0, 1, or 2. In certain embodiments, m is 0 or 1. In certain embodiments, m is 0. In other certain embodiments, m is 1.

[0488] Variable A is 0, 1, or 2. In certain embodiments, A is 0 or 1. In certain embodiments, A is 0.

[0489] Variable n is 0 or 1. In certain embodiments, n is 0. In other certain embodiments, n is 1.

[0490] In certain embodiments, A is 0; R a is hydrogen, C 1-6 aliphatic, C 1-6 fluoroaliphatic, or -(CH2) m -CH2-R B ; R a1 is hydrogen, C 1-6 aliphatic, C 1-6 fluoroaliphatic, -(CH2) m -CH2-R B , or -(CH2) m -CH(R 5a )-OR 5b ; P is R c -C(O)-, or R c -S(O)2-; R c is -R D ; and m is 0 or 1.

[0491] In certain other embodiments, A is 0; R a is C 1-6 aliphatic, or -(CH2) m -CH2-R B ; R a1 is hydrogen, -(CH2) m -CH2-R B , or -(CH2) m -CH(R 5a )-OR 5b ; P is R c -C(O)-, or R c -S(O)2-; R c is -R D ; and m is 0 or 1.

[0492] In certain other embodiments, A is 0; R a is C 1-6 aliphatic; R a1 is hydrogen, -(CH2) m -CH2-R B , or -(CH2) m -CH(R 5a )-OR 5b ; P is R c -C(O); R c is -RD ; and m is 0 or 1.

[0493] In certain other embodiments, A is 0; R a is isobutyl; R a1 is hydrogen, C 1-6 aliphatic, C 1-6 fluoroaliphatic, -(CH2) m -CH2-R B or -(CH2) m -CH(R 5a )-OR 5b ; P is R c -C(O)-; R c is -R D ; and m is 0 or 1.

[0494] In certain other embodiments, A is 0; R a is isobutyl; R a1 is hydrogen, C 1-6 aliphatic, -(CH2) m -CH2-R B or -(CH2) m -CH(R 5a )-OR 5b ; P is R c -C(O)-; R c is -R D ; and m is 0 or 1.

[0495] In certain other embodiments, A is 0; R a is isobutyl; R a1 is hydrogen, -(CH2) m -CH2-R B or -(CH2) m -CH(R 5a )-OR 5b ; P is R c -C(O)-; R c is -R D ; and m is 0 or 1.

[0496] In certain embodiments, A is 0; R a is isobutyl; R a1 is -CH2-R B , and R B is phenyl; P is R c -C(O)-; R c is -R D ; and R D is 2-pyrazinyl.

[0497] In certain other embodiments, A is 0; Ra is isobutyl; R a1 is hydrogen; P is R c -C(O)-; R c is -R D ; and R D is 2,5-dichlorophenyl.

[0498] In certain other embodiments, A is 0; R a is isobutyl; R a1 is -CH(R 5a )-OR 5b ; R 5a is C 1-6 aliphatic; R 5b is hydrogen; P is R c -C(O)-; R c is -R D ; and R D is 6-phenyl-2-pyridyl-.

[0499] In certain embodiments, the compound of Formula (I) or a crystalline form thereof is characterized by Formula (I-l):

[0500]

[0501] In certain other embodiments, the compound of Formula (I) or a crystalline form thereof is characterized by Formula (I-15):

[0502]

[0503] In certain other embodiments, the compound of Formula (I) or a crystalline form thereof is characterized by Formula (I-18):

[0504]

[0505] General Synthetic Methodology

[0506] Compounds of Formula (I) can be prepared by esterification of the corresponding boronic acid. The boronic acid compounds can be prepared by methods known to one of ordinary skill in the art. See, e.g., Adams et al., U.S. Patent No. 5,780,454; Pickersgill et al., International Patent Publication WO 2005 / 097809. An exemplary synthetic pathway is illustrated in Scheme 1 below.

[0507] Scheme 1:

[0508]

[0509] Compound i is coupled with N-protected amino acid ii, followed by N-terminal deprotection, to give compound iii or a salt thereof. Examples of suitable protecting groups (PG) include, but are not limited to, acyl protecting groups such as formyl, acetyl (Ac), succinyl (Suc), and methoxysuccinyl; and carbamate protecting groups such as t-butoxy carbonyl (Boc), benzyloxycarbonyl (Cbz), and fluorenylmethoxycarbonyl (Fmoc). Optionally, PG is hydrogen and deprotection is not required. The peptide coupling reaction can be carried out by pre-conversion of the carboxylic acid moiety of compound ii to an activated ester or acyl halide (e.g., O-(N-hydroxysuccinimidyl) ester), followed by treatment with compound i. Alternatively, the activated ester can be prepared in situ by contacting the carboxylic acid with a peptide coupling reagent. Examples of suitable peptide coupling reagents include, but are not limited to, carbodiimide reagents such as dicyclohexylcarbodiimide (DCC) or l-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC); phosphonium reagents such as (benzotriazol-l-yloxy)tris(dimethylamino)phosphonium hexafluorophosphate (BOP); and urea salt reagents such as O-(lH-benzotriazol-l-yl)-N,N,N',N'-tetramethyluronium tetrafluoroborate (TBTU).

[0510] Compound iii is then coupled with an amino-capping moiety to give compound iv. The peptide coupling conditions described above for the coupling of compounds i and ii are also suitable for the coupling of compound iii with the amino-capping moiety. Deprotection of the boronic acid moiety then gives compound v. The deprotection step is preferably carried out by treatment with a boronic acid acceptor in a hydrocarbon solvent and aqueous mineral acid. Examples of suitable boronic acid acceptors include, but are not limited to, BCl3, lithium aluminum hydride, and NaIO4. 5-8 The transesterification is effected in a biphasic mixture of a hydrocarbon solvent and aqueous mineral acid. Other reagents that can be used for the deprotection of the boronic acid moiety include, but are not limited to, BCl3, lithium aluminum hydride, and NaIO4.

[0511] Scheme 2:

[0512]

[0513] Alternatively, as shown in Scheme 2, the order of the coupling reactions can be reversed. Thus, O-protected amino acid vi is first coupled with the amino-capping moiety, followed by ester hydrolysis to form compound vii. Optionally, PG' is H and ester hydrolysis is not required, resulting directly in compound vii. Subsequent coupling with compound i and boronic acid deprotection to give compound v are then effected as described above for Scheme 1.

[0514] As shown in Scheme 3, compound v is reacted with a suitable α-hydroxy carboxylic acid or β-hydroxy carboxylic acid to give a compound of formula (I).

[0515] Scheme 3:

[0516]

[0517] The conversion of a compound of formula v to a compound of formula (I) can be achieved using about one molar equivalent of an a-hydroxy carboxylic acid or a β-hydroxy carboxylic acid, in a solvent such as ethyl acetate, at a temperature between about 40 °C and about 80 °C, under esterification conditions. The conversion of a compound of formula v to a compound of formula (I) can also be achieved using a molar excess of an a-hydroxy carboxylic acid or a β-hydroxy carboxylic acid as described above. Examples of other suitable solvents for this conversion include, but are not limited to, methyl isobutyl ketone, acetone, acetonitrile, 2-methyltetrahydrofuran, anisole, isopropyl acetate, dimethoxyethane, tetrahydrofuran, dioxane, dichloromethane, toluene, heptane, methyl-cyclohexane, t-butyl methyl ether, and mixtures thereof. The choice of solvent depends in part on the solubility of the a-hydroxy carboxylic acid or β-hydroxy carboxylic acid used. The temperature chosen for the conversion of a compound of formula v to a compound of formula (I) depends in part on the boiling point of the solvent or solvent mixture used.

[0518] The conversion of a compound of formula v to a compound of formula (I) can be catalyzed by an organic amine base such as, but not limited to, triethylamine, triethylenediamine, pyridine, collidine, 2,6-lutidine, 4-dimethylaminopyridine, di-tert-butylpyridine, N-methylmorpholine, N-methylpiperidine, tetramethylguanidine, diazabicyclo[5.4.0]undec-7-ene (DBU), 1,4-diazabicyclo[2.2.2]octane, 1,5-diazabicyclo[4.3.0]non-5-ene, N,N'-diisopropylethylamine, or mixtures thereof.

[0519] The compound of formula v and the a-hydroxy carboxylic acid or β-hydroxy carboxylic acid are heated together in the selected solvent for a period of time. After this period of time, the reaction mixture is allowed to cool for a period of time and the compound of formula (I) that precipitates upon cooling is collected by filtration. The cooling can be uncontrolled or it can be controlled by the use of a cooling device. The reaction mixture can be stirred during this cooling period. Alternatively, the compound of formula (I) can also be isolated from the reaction mixture by cooling followed by evaporation of the solvent. The reaction mixture can be seeded with crystals of the compound of formula (I) in order to effect precipitation.

[0520] A co-solvent such as, but not limited to, heptane, methylcyclohexane, toluene, t-butyl methyl ether, ethyl acetate, or mixtures thereof can be added during the cooling period. After the addition of the co-solvent, the reaction mixture can be further cooled, causing the compound of formula (I) to precipitate. Alternatively, once the co-solvent is added, the reaction mixture can be subsequently reheated to produce a homogeneous solution, which is then cooled, causing the compound of formula (I) to precipitate. The reaction mixture can be seeded with crystals of the compound of formula (I) in order to effect precipitation.

[0521] In other embodiments, the compound of formula (I) is isolated in substantially pure form. In such embodiments, the purity is about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or about 99.5%.

[0522] In certain embodiments, the compound of formula (I) is isolated in crystalline form. In certain embodiments, the compound of formula (I) is isolated in substantially crystalline form. In certain other embodiments, the compound of formula (I) is isolated in amorphous form.

[0523] The compound of formula (I) can also be co-lyophilized by co-lyophilizing compound v with an alpha-hydroxy carboxylic acid or a beta-hydroxy carboxylic acid. This is accomplished by subjecting an aqueous solution comprising the compound of formula v and a molar excess of an alpha-hydroxy carboxylic acid or a beta-hydroxy carboxylic acid to a lyophilization procedure. In certain embodiments, the aqueous solution additionally comprises a water-miscible co-solvent. Examples of suitable co-solvents include, but are not limited to, t-butanol, methanol, ethanol, and mixtures thereof. Co-lyophilization results in a composition containing the compound of formula (I) and the excess alpha-hydroxy carboxylic acid or beta-hydroxy carboxylic acid.

[0524] Uses, formulations, and administration

[0525] The present application provides compounds that are effective inhibitors of proteasomes. The ability of a compound to inhibit proteasome-mediated hydrolysis of peptides or degradation of proteins can be assayed in vitro or in vivo.

[0526] Thus, in another aspect, the present application provides a method of inhibiting one or more peptidase activities of a proteasome in a cell, comprising contacting the cell in which proteasome inhibition is desired with a compound described herein, or a pharmaceutically acceptable salt, borate, or boronic anhydride thereof.

[0527] The present application also provides a method of inhibiting cell proliferation, comprising contacting a cell in which such inhibition is desired with a compound described herein. The phrase "inhibiting cell proliferation" is used to mean the ability of a compound of the present application to inhibit the number of cells or the growth of cells in a contacted cell as compared to a cell that has not been contacted with an inhibitor. Cell proliferation can be assessed by counting cells using a cell counter or by a cell viability assay, such as an MTT or WST assay. In the case where cells are in a solid growth, such as a solid tumor or an organ, such assessment of cell proliferation can be performed by, for example, measuring growth with a caliper and comparing the size of growth of contacted cells to uncontacted cells.

[0528] Preferably, the growth of the cells contacted with the inhibitor is retarded by at least about 50% as compared to the growth of uncontacted cells. In various embodiments, the cell proliferation of the contacted cells is inhibited by at least about 75%, at least about 90%, or at least about 95% as compared to uncontacted cells. In certain embodiments, the phrase "inhibits cell proliferation" includes a reduction in the number of cells contacted as compared to uncontacted cells. Thus, a proteasome inhibitor that inhibits cell proliferation of contacted cells can induce the contacted cells to undergo growth retardation, to undergo growth arrest, to undergo progressive cell death (i.e., apoptosis), or to undergo necrotic cell death.

[0529] In another aspect, the present application provides a pharmaceutical composition comprising a compound of Formula (I), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0530] In certain embodiments, the composition further comprises a free alpha-hydroxy carboxylic acid or salt thereof or beta-hydroxy carboxylic acid or salt thereof. In such embodiments, the alpha-hydroxy carboxylic acid or salt thereof or beta-hydroxy carboxylic acid or salt thereof is present in a molar ratio to the compound of Formula (I) in the range of about 2: 1 to about 200: 1. In various embodiments, the alpha-hydroxy carboxylic acid or salt thereof or beta-hydroxy carboxylic acid or salt thereof is present in a molar ratio to the compound of Formula (I) in the range of about 2: 1 to about 200: 1, about 15: 1 to about 80: 1, or about 20: 1 to about 40: 1.

[0531] If pharmaceutically acceptable salts of the compounds of the present application are used in these compositions, the salts are preferably derived from inorganic or organic acids or bases. For a review on suitable salts, see, e.g., Berge et al., J. Pharm. Sci. 66:1-19 (1977) and Remington: The Science and Practice of Pharmacy, 20thEdition, Edited by A. Gennaro, Lippincott Williams & Wilkins, 2000.

[0532] Non-limiting examples of suitable acid addition salts include the following: acetate, adipate, alginate, aspartate, benzoate, besylate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptanoate, glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, oxalate, pamoate, pectinate, persulfate, 3-phenylpropionate, picrate, pivalate, propionate, succinate, tartrate, thiocyanate, tosylate, and undecanoate.

[0533] Suitable base addition salts include, but are not limited to, ammonium salts; alkali metal salts, e.g., lithium, sodium and potassium salts; alkaline earth metal salts, e.g., calcium and magnesium salts; other polyvalent metal salts, e.g., zinc salts; salts with organic bases, e.g., dicyclohexylamine, N-methyl-D-glucamine, t-butylamine, ethylenediamine, ethanolamine and choline; and salts with amino acids, e.g., arginine, lysine, and the like.

[0534] The term "pharmaceutically acceptable carrier" is used herein to refer to a material that is compatible with the individual subject (preferably a mammal, more preferably a human) to whom the active agent is being delivered and is suitable for delivery of the active agent to the target site without terminating the activity of the agent. Toxicity or adverse effects associated with the carrier, if any, are preferably commensurate with a reasonable risk / benefit ratio for the intended use of the active agent.

[0535] The terms "carrier," "excipient," or "vehicle" are used interchangeably herein and include any and all solvents, diluents, and other liquid vehicles, dispersion or suspension aids, surface active agents, pH adjusting agents, isotonic agents, thickening or emulsifying agents, preservatives, solid binders, lubricants, and the like, as suited to the particular dosage form desired. Remington: The Science and Practice of Pharmacy, 20th Edition, A. Gennaro, Lippincott Williams & Wilkins, 2000 discloses various carriers used in formulating pharmaceutically acceptable compositions and known techniques for the preparation thereof. Strickley, Pharmaceutical Research, 21(2) 201-230 (2004) reviews pharmaceutically acceptable excipients useful for dissolving compounds for oral or parenteral administration in commercial products. Except insofar as any conventional carrier medium is incompatible with a compound of the application, such as by producing an undesirable biological effect or otherwise interacting in a deleterious manner with any other component(s) of the pharmaceutically acceptable composition, its use is contemplated to be within the scope of this application. Certain examples of materials that can serve as pharmaceutically acceptable carriers include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins (such as human serum albumin), buffer substances (such as phosphates, carbonates, magnesium trisilicate and aluminum hydroxide), glycine, sorbic acid or sorbitan, partially glyceride mixtures of saturated vegetable fatty acids, water, physiologically saline, protamine sulfate, dactylamine phosphate and zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, polyacrylates, waxes, polyethylene-polyoxypropylene- block polymers, wool fat, sugars (such as lactose, glucose, sucrose, and mannitol), starches (such as corn starch and potato starch), cellulose and its derivatives (such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate), powdered tragacanth, malt, gelatin, talc, excipients (such as cocoa butter and suppository waxes), oils (such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil), glycols (such as propylene glycol and polyethylene glycol), esters (such as ethyl oleate and ethyl laurate), agar, alginic acid, isotonic saline, Ringer's solution, alcohols (such as ethyl alcohol, isopropyl alcohol, cetyl alcohol and glycerol), cyclodextrins (such as hydroxypropyl beta-cyclodextrin and sulfobutyl ether beta-cyclodextrin), lubricants (such as sodium lauryl sulfate and magnesium stearate), petroleum hydrocarbons (such as mineral oil and petrolatum). Coloring agents, releasing agents, coating agents, sweetening agents, flavoring agents and perfuming agents, preservatives and antioxidants can also be present in the composition, according to the judgment of the formulator.

[0536] The pharmaceutical compositions of the present application can be manufactured in a manner appropriate to the type of composition desired, for example, by conventional methods of granulation, mixing, dissolving, encapsulation, lyophilization, or emulsification. The composition can take the form of granules, powders, or micro particles, a powder (including a freeze-dried, spray-dried or spray freeze-dried powder, an amorphous powder), tablets, capsules, syrups, suppositories, injections, emulsions, elixirs, suspensions or solutions.

[0537] According to one preferred embodiment, the compositions of the present application are formulated for pharmaceutical administration to mammals, preferably humans. The pharmaceutical compositions of the present application can be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally, or via an implanted reservoir. The term "parenterally" 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, intravenously, or subcutaneously. The formulations of the present application can be designed to be short-acting, fast-releasing, or long-acting. In addition, the compounds can be administered in a local rather than systemic fashion, for example, via injection into a tumor (e.g., by intralesional injection).

[0538] Oral liquid dosage forms include, but are not limited to, pharmaceutically-acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to the active compounds, the liquid dosage forms can contain inert diluents commonly used in the art, such as water or other solvents, solubilizing agents and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3- butylene glycol, cyclodextrins, dimethylformamide, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof. Besides inert diluents, the oral compositions can also include adjuvants, such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0539] Injectable preparations (e.g., aqueous or oleaginous sterile injectable suspensions) can be formulated according to known techniques using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation can also be a sterile injectable solution, suspension or emulsion 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 can be employed are water, Ringer's solution, U.S.P. and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil can be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables. The injectable formulations can be sterilized, for example, by filtration through a bacterial-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use. Compositions for parenteral administration can be injected by bolus injection or by continuous infusion, or can be administered by continuous infusion.

[0540] Oral solid dosage forms include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound is admixed with at least one inert, pharmaceutically-acceptable excipient or carrier, such as sodium citrate or dicalcium phosphate and / or a) fillers or extenders, such as starches, lactose, sucrose, cellulose, dextrose, mannitol, and silicic acid; b) binders, such as carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and acacia; c) humectants, such as glycerol; d) disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, croscarmellose sodium, corn starch, sodium starch glycolate, and sodium carbonate; e) solution retarders, such as paraffin; f) absorption accelerators, such as quaternary ammonium compounds; g) wetting agents, such as cetyl alcohol and glycerol monostearate; h) absorbents, such as kaolin and bentonite clay; and i) lubricants, such as talc, calcium stearate, magnesium stearate, sodium stearate, zinc stearate, stearic acid, solid polyethylene glycols, sodium lauryl sulfate, glyceryl monostearate, and mixtures thereof. In the case of capsules, tablets, and pills, the dosage forms can also comprise buffering agents, such as a phosphate or citrate.

[0541] Solid compositions of a similar type can also be employed as fillers in soft and hard filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols, and the like. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical formulating art. They can optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. Solid compositions of a similar type can also be employed as fillers in soft and hard filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polethylene glycols, and the like.

[0542] The active compounds can also be in micro-encapsulated form, with one or more excipients as noted above. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings, release control coatings and other coatings well known in the pharmaceutical formulating art. In such solid dosage forms the active compound can be admixed with at least one inert diluent such as sucrose, lactose or starch. Such dosage forms can also comprise, as is normal practice, additional substances other than inert diluents, e.g., tableting lubricants and other tableting aids, such a magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and pills, dosage forms can also comprise buffering agents. They can optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. In certain embodiments, excipients or carriers can include, but are not limited to, sodium stearyl fumarate, carboxymethylcellulose, magnesium stearate, cross-linked polyvinyl pyrrolidone, ethyl cellulose, talc, and silicified microcrystalline cellulose.

[0543] Dosage forms for topical or transdermal administration of a compound of this application include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants, or patches. The active component is admixed under sterile conditions with a pharmaceutically acceptable carrier and any needed preservatives or buffers as can be required. Ophthalmic formulation, eardrops, and eye drops are also contemplated as being within the scope of this application. Additionally, the present application contemplates the use of transdermal patches, which have the added advantage of providing controlled delivery of a compound to the body. Such dosage forms can be made by dissolving or dispensing the compound in the proper medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate of such flux can be controlled by either providing a rate controlling membrane or dispersing into polymer matrices or gels.

[0544] In certain embodiments, the present application provides a pharmaceutical composition comprising a compound of Formula (I) and additional excipients described herein. In certain other embodiments, the present application provides a pharmaceutical composition comprising a compound of Formula (II) and additional excipients described herein. In certain other embodiments, the present application provides a pharmaceutical composition comprising a compound of Formula (III) or (IV) and additional excipients described herein.

[0545] In other embodiments, the present application provides a pharmaceutical composition comprising a citric acid ester of compound (VIII-1) and additional excipients described herein. In other embodiments, the present application provides a pharmaceutical composition comprising a citric acid ester of compound (VIII-15) and additional excipients described herein. In other embodiments, the present application provides a pharmaceutical composition comprising a citric acid ester of compound (VIII-18) and additional excipients described herein.

[0546]

[0547] In other embodiments, the present application provides a pharmaceutical composition comprising a compound of Formula (I-1) or a crystalline form thereof. In other embodiments, the present application provides a pharmaceutical composition comprising a compound of Formula (I-15) or a crystalline form thereof. In certain other embodiments, the present application provides a pharmaceutical composition comprising a compound of Formula (I-18) or a crystalline form thereof.

[0548] The following description of pharmaceutical compositions and methods of making the pharmaceutical compositions apply to compounds of Formula (I), (II), (III), (Ilia), (IV), or (IVa), and various embodiments of these formulas, as described herein. The following description of pharmaceutical compositions and methods of making the pharmaceutical compositions also apply to compounds (I-1), (I-15), or (I-18).

[0549] In one embodiment, the pharmaceutical composition comprises a compound of Formula (I), wherein the compound of Formula (I) is substantially crystalline. In another embodiment, the compound of Formula (I) in the pharmaceutical composition is at least about 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9% crystalline. In another embodiment, the compound of Formula (I) in the pharmaceutical composition is crystalline.

[0550] In certain embodiments, the pharmaceutical formulations of the present application provide stable solid oral dosage forms of the active compound by using excipients with low water or low moisture content and using dry or anhydrous formulation methods of manufacture.

[0551] In one embodiment, the pharmaceutical composition is an oral pharmaceutical dosage form selected from the group consisting of capsules, tablets, pills, powders, and granules. In another embodiment, the oral pharmaceutical dosage form is a capsule, wherein the capsule is a polymer-based capsule selected from the group consisting of gelatin, hydroxypropyl methylcellulose (HPMC), fish gelatin, and pullulan. In another embodiment, the polymer-based capsule is selected from the group consisting of gelatin and hydroxypropyl methylcellulose. In another embodiment, the polymer-based capsule is a hard gelatin capsule.

[0552] In one embodiment, the pharmaceutical composition comprises the compound of Formula (I) or a crystalline form thereof, a filler, and optionally a lubricant. In another embodiment, the pharmaceutical composition comprises about 0.2% to about 3% of the compound of Formula (I) or a crystalline form thereof; about 97% to about 99.8% of a filler; and optionally up to about 1.5% of a lubricant. In another embodiment, the pharmaceutical composition comprises about 0.25% to about 2% of the compound of Formula (I) or a crystalline form thereof; and about 98% to about 99.75% of a filler.

[0553] In another embodiment, the pharmaceutical composition further comprises an optional glidant and an optional buffer. In another embodiment, the pharmaceutical composition comprises, in weight percent of total weight, about 0.2% to about 3% of the compound of Formula (I) or a crystalline form thereof, about 86.5% to about 99.8% of a filler, optionally up to about 1.5% of a lubricant, optionally up to about 5% of a glidant, and optionally up to about 5% of a buffer.

[0554] In another embodiment, the pharmaceutical composition comprises, in weight percent of total weight, about 0.2% to about 12% of the compound of Formula (I) or a crystalline form thereof, about 76.5% to about 99.8% of a filler, optionally up to about 1.5% of a lubricant, optionally up to about 5% of a glidant, and optionally up to about 5% of a buffer.

[0555] In certain embodiments, the compound of Formula (I) or a crystalline form thereof is present in the pharmaceutical composition in an amount of about 0.2% to about 3% by weight percent of total weight. In certain other embodiments, the compound of Formula (I) or a crystalline form thereof is present in the pharmaceutical composition in an amount of about 0.25% to about 2% by weight percent of total weight.

[0556] Suitable fillers include, but are not limited to, powdered cellulose, microcrystalline cellulose, silicified microcrystalline cellulose, high-density microcrystalline cellulose, low moisture microcrystalline cellulose, pregelatinized starch, sodium starch glycolate, and mixtures thereof. In certain other embodiments, the filler is selected from the group consisting of powdered cellulose, microcrystalline cellulose, silicified microcrystalline cellulose, low moisture microcrystalline cellulose, and mixtures thereof. In certain other embodiments, the filler is low moisture microcrystalline cellulose. In certain other embodiments, the filler is selected from the group consisting of low moisture microcrystalline cellulose, sodium starch glycolate, pregelatinized starch, and mixtures thereof.

[0557] In other embodiments, the filler is present in an amount of about 97% to about 99.8% by weight based on total weight. In certain other embodiments, the filler is present in an amount of about 98% to about 99.75% by weight based on total weight. In certain other embodiments, when a lubricant is present, the filler is reduced in an amount corresponding to the percentage of lubricant present. In certain other embodiments, the filler is present in an amount of about 86.5% to about 99.8% by weight based on total weight.

[0558] In certain embodiments, the filler comprises a first filler and a second filler. As long as the total amount of filler is no more than about 99.8%, the first filler is present in an amount of 0% to about 99.8% by weight based on total weight, and the second filler is present in an amount of 0% to about 99.8% by weight based on total weight. In certain embodiments, as long as the total amount of filler is no more than about 99.8% by weight based on total weight, the first filler is present in an amount of about 40% to about 60% by weight based on total weight, and the second filler is present in an amount of about 40% to about 60% by weight based on total weight.

[0559] In certain embodiments, the first filler is selected from the group consisting of low moisture microcrystalline cellulose, sodium starch glycolate, pregelatinized starch, and mixtures thereof. In certain embodiments, the second filler is selected from the group consisting of low moisture microcrystalline cellulose, sodium starch glycolate, pregelatinized starch, and mixtures thereof.

[0560] Suitable lubricants include, but are not limited to, magnesium stearate, glyceryl behenate, hydrogenated vegetable oil, talc, zinc stearate, calcium stearate, sucrose stearate, sodium stearyl fumarate, and mixtures thereof. In certain embodiments, the lubricant is magnesium stearate. In other embodiments, the lubricant is present in an amount of up to about 1.5% by weight based on total weight. In certain other embodiments, the lubricant is present in an amount of about 1% by weight based on total weight.

[0561] Suitable glidants include, but are not limited to, silicon dioxide, talc, and mixtures thereof. In certain embodiments, the glidant is talc. In other embodiments, the glidant is present in an amount of up to about 5% by weight of the total weight. In certain other embodiments, the glidant is present in an amount of about 1% by weight of the total weight. In certain other embodiments, the glidant is present in an amount of about 2% by weight of the total weight.

[0562] Suitable buffers include, but are not limited to, sodium citrate, citric acid, and mixtures thereof. In certain embodiments, the buffer is sodium citrate. In certain other embodiments, the buffer is present in an amount of up to about 5% by weight of the total weight. In certain other embodiments, the buffer is present in an amount of about 2% by weight of the total weight.

[0563] In certain embodiments, the pharmaceutical composition comprises a compound of Formula (I) or a crystalline form thereof, a filler, and optionally a lubricant; wherein:

[0564] the a-hydroxy carboxylic acid or the β-hydroxy carboxylic acid is citric acid;

[0565] A is 0;

[0566] R a is isobutyl;

[0567] R a1 is hydrogen, C 1-6 aliphatic, -(CH2) m -CH2-R B or -(CH2) m -CH(R 5a )-OR 5b ;

[0568] P is R c -C(O)-;

[0569] R c is -R D ;

[0570] m is 0 or 1;

[0571] the filler is selected from the group consisting of low moisture microcrystalline cellulose, sodium starch glycolate, pregelatinized starch, and mixtures thereof; and

[0572] the lubricant, when present, is magnesium stearate.

[0573] In certain embodiments, the pharmaceutical composition comprises a compound of Formula (I) or a crystalline form thereof, a filler, and optionally a lubricant; wherein:

[0574] the compound of Formula (I) is (I-1), (I-15), or (I-18);

[0575] The filler is selected from the group consisting of low moisture microcrystalline cellulose, sodium starch glycolate, pregelatinized starch, and mixtures thereof; and

[0576] The lubricant, when present, is magnesium stearate.

[0577] In certain embodiments, the pharmaceutical composition comprises about 0.25% to about 2% of the compound of Formula (I) or a crystalline form thereof; and about 98% to about 99.75% of a filler; wherein:

[0578] The compound of Formula (I) is (I-1), (I-15), or (I-18); and

[0579] The filler is selected from the group consisting of low moisture microcrystalline cellulose, sodium starch glycolate, pregelatinized starch, and mixtures thereof.

[0580] In certain embodiments, the pharmaceutical composition comprises the compound of Formula (I) or a crystalline form thereof, a filler, optionally a lubricant, optionally a glidant, and optionally a buffer; wherein:

[0581] The a-hydroxycarboxylic acid or β-hydroxycarboxylic acid is citric acid;

[0582] A is 0;

[0583] R a is isobutyl;

[0584] R a1 is hydrogen, C 1-6 aliphatic, -(CH2) m -CH2-R B or -(CH2) m -CH(R 5a )-OR 5b ;

[0585] P is R c -C(O)-;

[0586] R c is -R D ;

[0587] m is 0 or 1;

[0588] The filler is selected from the group consisting of low moisture microcrystalline cellulose, sodium starch glycolate, pregelatinized starch, and mixtures thereof.

[0589] The lubricant, when present, is magnesium stearate;

[0590] The glidant, when present, is talc; and

[0591] The buffer, when present, is sodium citrate.

[0592] In some embodiments, the pharmaceutical composition comprises a compound of formula (I) or its crystalline form, a filler, an optional lubricant, an optional flow aid, and an optional buffer, wherein:

[0593] The compounds of formula (I) are (I-1), (I-15), or (I-18);

[0594] The filler is selected from the group consisting of low-moisture microcrystalline cellulose, sodium glycolate starch, pregelatinized starch and mixtures thereof;

[0595] The lubricant, when present, is magnesium stearate;

[0596] The gliding agent is talc when present; and

[0597] The buffer, when present, is sodium citrate.

[0598] In some embodiments, the pharmaceutical composition comprises, by weight percentage of the total weight, about 0.2% to about 3% of a compound of formula (I) or its crystalline form, about 86.5% to about 99.8% of a filler, optionally up to about 1.5% of a lubricant, optionally up to about 5% of a flow aid, and optionally up to about 5% of a buffer, wherein:

[0599] The compounds of formula (I) are (I-1), (I-15), or (I-18);

[0600] The filler is selected from the group consisting of low-moisture microcrystalline cellulose, sodium glycolate starch, pregelatinized starch and mixtures thereof;

[0601] The lubricant, when present, is magnesium stearate;

[0602] The gliding agent is talc when present; and

[0603] The buffer, when present, is sodium citrate.

[0604] In some embodiments, the pharmaceutical composition comprises a compound of formula (I) or its crystalline form, a filler, and optionally a lubricant; wherein the compound of formula (I) is formula (I-1). In some other embodiments, the pharmaceutical composition comprises a compound of formula (I) or its crystalline form, a filler, and optionally a lubricant; wherein the compound of formula (I) is formula (I-1); the filler is selected from the group consisting of low-moisture microcrystalline cellulose, sodium glycolate starch, pregelatinized starch, and mixtures thereof; and the lubricant, when present, is magnesium stearate.

[0605] In some embodiments, the pharmaceutical composition comprises a compound of formula (I) or its crystalline form; wherein the compound of formula (I) is (I-1); and the crystalline form is form 2.

[0606] In certain embodiments, the pharmaceutical composition comprises Compound (I-1) Form 2 and low moisture microcrystalline cellulose. In certain other embodiments, the pharmaceutical composition comprises Compound (I-1) Form 2 and siliconized microcrystalline cellulose. In certain other embodiments, the pharmaceutical composition comprises Compound (I-1) Form 2, low moisture microcrystalline cellulose, and magnesium stearate. In certain other embodiments, the pharmaceutical composition comprises Compound (I-1) Form 2, microcrystalline cellulose, and magnesium stearate.

[0607] In certain embodiments, the pharmaceutical composition comprises Compound (I-1) Form 2, low moisture microcrystalline cellulose, and talc. In certain other embodiments, the pharmaceutical composition comprises Compound (I-1) Form 2 and pregelatinized starch. In certain other embodiments, the pharmaceutical composition comprises Compound (I-1) Form 2, pregelatinized starch, talc, and magnesium stearate. In certain other embodiments, the pharmaceutical composition comprises Compound (I-1) Form 2, low moisture microcrystalline cellulose, talc, and magnesium stearate. In certain other embodiments, the pharmaceutical composition comprises Compound (I-1) Form 2, low moisture microcrystalline cellulose, talc, magnesium stearate, and sodium citrate. In certain other embodiments, the pharmaceutical composition comprises Compound (I-1) Form 2, low moisture microcrystalline cellulose, talc, magnesium stearate, and pregelatinized starch. In certain other embodiments, the pharmaceutical composition comprises Compound (I-1) Form 2, low moisture microcrystalline cellulose, talc, magnesium stearate, and sodium starch glycolate.

[0608] When a compound of Formula (I) is subjected to hydrolytic conditions, the ester moiety of the molecule hydrolyzes to give a compound of Formula (VIII) in a 1 : 1 molar ratio.

[0609]

[0610] The amount of compound of Formula (VIII) present in the sample is measured using an analytical method that involves hydrolytic conditions on the sample preparation by comparison to a reference standard of known purity (see, e.g., Analytical Test Method 1 below). The amount of compound of Formula (VIII) present in a sample of a compound of Formula (I) is measured using an analytical method that does not subject the sample to hydrolytic conditions by comparison to a reference standard of known purity (see, e.g., Analytical Test Method 2 below). Thus, the amount of compound of Formula (VIII) measured in Analytical Test Method 1 minus the amount of compound of Formula (VIII) measured in Analytical Test Method 2 gives the amount of compound of Formula (VIII) in the sample that has been derived via hydrolysis of a compound of Formula (I). The amount of compound of Formula (I) present in the sample is obtained based on a 1 : 1 molar ratio conversion of a compound of Formula (I) to a compound of Formula (VIII) with a molecular weight conversion.

[0611] It will be appreciated that the analytical methods described directly above and in the Experimental Section below apply in a similar manner to any of the compounds of Formula (I), (II), (III), (Ilia), (IV), or (IVa), and the various embodiments of these formulas, as described herein. The analytical methods described directly above and in the Experimental Section below apply in a similar manner to compounds (I-1), (I-15), or (I-18).

[0612] In certain embodiments, the amount of the compound of Formula (VIII) present in the pharmaceutical composition is determined by measuring the amount of the compound of Formula (VIII) present after subjecting a test sample to conditions that hydrolyze the compound of Formula (I) to the compound of Formula (VIII).

[0613] In certain embodiments, the amount of the compound of Formula (I-1), or a crystalline form thereof, present in the pharmaceutical composition is expressed in an amount equivalent to the molar weight of the compound of Formula (VIII-1).

[0614] In certain embodiments, the present application relates to a unit dose pharmaceutical composition comprising a compound of Formula (I-1), or a crystalline form thereof.

[0615] In certain other embodiments, the unit dose pharmaceutical composition comprises a compound of Formula (I-1), or a crystalline form thereof, wherein the compound of Formula (I-1) is present in an amount equivalent to about 0.1 mg to about 3.0 mg of the molar weight of the compound of Formula (VIII-1). In certain other embodiments, the unit dose pharmaceutical composition comprises a compound of Formula (I-1), or a crystalline form thereof, wherein the compound of Formula (I-1) is present in an amount equivalent to about 0.15 mg to about 2.2 mg of the molar weight of the compound of Formula (VIII-1). In certain other embodiments, the unit dose pharmaceutical composition comprises a compound of Formula (I-1), or a crystalline form thereof, wherein the compound of Formula (I-1) is present in an amount equivalent to about 0.18 mg to about 0.22 mg of the molar weight of the compound of Formula (VIII-1). In certain other embodiments, the unit dose pharmaceutical composition comprises a compound of Formula (I-1), or a crystalline form thereof, wherein the compound of Formula (I-1) is present in an amount equivalent to about 0.46 mg to about 0.54 mg of the molar weight of the compound of Formula (VIII-1). In certain other embodiments, the unit dose pharmaceutical composition comprises a compound of Formula (I-1), or a crystalline form thereof, wherein the compound of Formula (I-1) is present in an amount equivalent to about 1.80 mg to about 2.20 mg of the molar weight of the compound of Formula (VIII-1).

[0616] In certain embodiments, the amount of the compound of Formula (I-1), or a crystalline form thereof, present in the pharmaceutical composition is expressed in an equivalent amount of the compound of Formula (VIII-1), based on the relative molecular weights of the compound of Formula (I-1) and the compound of Formula (VIII-1).

[0617] In certain embodiments, the unit dose pharmaceutical composition comprises about 0.143 mg to about 4.3 mg of the compound of formula (I-1), or a crystalline form thereof, based on a weight ratio, measured as about 0.1 mg to about 3.0 mg of the compound of formula (VIII-1).

[0618] In certain other embodiments, the unit dose pharmaceutical composition comprises about 0.214 mg to about 3.15 mg of the compound of formula (I-1), or a crystalline form thereof, based on a weight ratio, measured as about 0.15 mg to about 2.2 mg of the compound of formula (VIII-1).

[0619] In certain other embodiments, the unit dose pharmaceutical composition comprises about 0.258 mg to about 0.315 mg of the compound of formula (I-1), or a crystalline form thereof, based on a weight ratio, measured as about 0.18 mg to about 0.22 mg of the compound of formula (VIII-1).

[0620] In certain other embodiments, the unit dose pharmaceutical composition comprises about 0.659 mg to about 0.773 mg of the compound of formula (I-1), or a crystalline form thereof, based on a weight ratio, measured as about 0.46 mg to about 0.54 mg of the compound of formula (VIII-1).

[0621] In certain other embodiments, the unit dose pharmaceutical composition comprises about 2.58 mg to about 3.15 mg of the compound of formula (I-1), or a crystalline form thereof, based on a weight ratio, measured as about 1.80 mg to about 2.20 mg of the compound of formula (VIII-1).

[0622] In certain embodiments, the present application provides a method of preparing an oral pharmaceutical dosage form of a compound of formula (I), or a crystalline form thereof, wherein the oral pharmaceutical dosage form is a capsule, the method comprising the steps of:

[0623] (a-1) mixing together a sieved filler and a sieved compound of formula (I), or a crystalline form thereof, in a bag;

[0624] (a-2) passing the mixture resulting from step (a-1) through a screen followed by blending;

[0625] (a-3) sieving additional filler through the same screen, passing it through the same bag, and blending in the same blending device;

[0626] (a-4) repeating step (a-3) up to two times;

[0627] (a-5) obtaining the mixture resulting from step (a-4) and encapsulating it using a capsule filling system; and

[0628] (a-6) weight picking the capsules resulting from step (a-5).

[0629] In certain embodiments, step (a-3) can be repeated three or more times.

[0630] When a lubricant is present in the pharmaceutical composition, the present application provides a method of preparing an oral pharmaceutical dosage form of a compound of formula (I) or a crystalline form thereof, wherein the oral pharmaceutical dosage form is a capsule, the method comprising the steps of:

[0631] (b-1) mixing together the sieved filler and the sieved compound of formula (I) or a crystalline form thereof in a bag;

[0632] (b-2) passing the mixture resulting from step (b-1) through a screen followed by blending;

[0633] (b-3) sieving additional filler through the same screen, passing it through the same bag, and blending in the same blending device;

[0634] (b-4) repeating step (b-3) up to two times;

[0635] (b-5) blending together the mixture from step (b-4) and the sieved lubricant;

[0636] (b-6) obtaining the mixture resulting from step (b-5) and encapsulating it using a capsule filling system; and

[0637] (b-7) weight picking the capsules resulting from step (b-6).

[0638] In certain embodiments, step (b-3) can be repeated three or more times. When additional components (e.g. a buffer, a second filler, or a glidant) are present in the pharmaceutical composition, they can be added in either of steps (b-1) or (b-3). The total amount of each component in the pharmaceutical composition can be added in one step or can be divided into several amounts, which can or can not be equal, and added in the individually occurring steps (b-1) or (b-3).

[0639] In certain embodiments, the present application provides a method of preparing an oral pharmaceutical dosage form of a compound of formula (I) or a crystalline form thereof, wherein the oral pharmaceutical dosage form is a capsule, the method comprising the steps of:

[0640] (c-1) passing the filler through a screen followed by placing it in a high shear mixing device;

[0641] (c-2) passing the compound of formula (I) or a crystalline form thereof through a screen followed by placing it in the same high shear mixing device;

[0642] (c-3) passing the filler through a screen followed by placing it in the same high shear mixing device;

[0643] (c-4) mixing using the same high shear mixing device for less than 10 minutes;

[0644] (c-5) obtaining the mixture from step (c-4) and encapsulating it using a capsule filling system; and

[0645] (c-6) sieving the capsules from step (c-5) by weight.

[0646] In certain embodiments, when a high shear mixing device is used, additional components present in the pharmaceutical composition can be added by repeating step (c-1) or step (c-3).

[0647] In certain embodiments, the compound of formula (I) used in the above described method of preparing a solid oral dosage form is selected from the group consisting of (I-1), (I-15), and (I-18). In certain embodiments, the compound of formula (I) used in the above described method of preparing a solid oral dosage form is (I-1).

[0648] The above described method steps can be performed using conventional devices and equipment. For a review, see, e.g., Remington: The Science and Practice of Pharmacy, 21stEd., Lippincott Williams & Wilkins, 2005.

[0649] The above described blending steps can be performed in any conventional blending device. In certain embodiments, the blending time for each individual blending step is between about 1 minute and about 45 minutes. In certain other embodiments, the blending time for each individual blending step is between about 1 minute and about 20 minutes. In certain other embodiments, the blending time for each individual blending step is between about 2 minutes and about 15 minutes.

[0650] The above described mixing steps can be performed in any conventional polyethylene bag. In certain embodiments, the mixing step is performed for between about 30 seconds and 5 minutes. In certain embodiments, the above described mixing steps can be performed in a stainless steel container.

[0651] The mixing step using a high shear mixing device can be performed in any conventional high shear mixing device. One example of such a high shear mixing device is sold as a Lab High Shear Granulator (Key International, Inc., Englishtown, NJ). In certain embodiments, the mixing is performed for less than about 10 minutes. In certain other embodiments, the mixing is performed for less than about 5 minutes.

[0652] The capsule filling step described above can be performed in any conventional filling system or device. In certain embodiments, the capsule filling system is a semi-automated filling system and can handle small batches. One example of such a capsule filling system is sold by In-Cap (Isopak Limited, Lincolnshire, Stamford, United Kingdom). In certain embodiments, the capsule filling system is a manual filling system. One example of such a capsule filling device is sold by ProFill 100 (torpac, Inc., Fairfield, NJ, USA).

[0653] In certain embodiments, the capsule is a hard gelatin capsule sold by Capsugel (Capsugel, Peapack, NJ). One skilled in the art can select the appropriate capsule size and color. In certain embodiments, the capsule has a fill weight of 85 mg, 120 mg, or 150 mg.

[0654] The weight sorting step described above can be performed using any conventional weight sorting device or machine. One example of a weight sorting device or machine is sold by SADE SP Bench Top Tablet and Capsule Weight Sorter (AC Compacting LLC, North Brunswick, NJ, USA).

[0655] In certain embodiments, the capsules are packaged in a bottle, a foil pouch, or a blister pack. In certain other embodiments, the capsules are packaged in a heat- induced sealed high-density polyethylene (HDPE) bottle. In another embodiment, the capsules are packaged in an air-tight sealed foil pouch. In another embodiment, the capsules are packaged in a foil-foil blister pack. In certain other embodiments, the capsules are packaged with a desiccant.

[0656] The physical and chemical stability of the oral pharmaceutical dosage form can be tested in a conventional manner, for example, by measuring the extent of dissolution, the time to disintegration, and determining the degradation products of the compound of Formula (I) after storage at different temperatures for different durations.

[0657] In certain other embodiments, the present application provides a pharmaceutical composition for non-parenteral use. In certain other embodiments, the present application provides a liquid pharmaceutical composition for non-parenteral or oral use.

[0658] In certain embodiments, the compound of formula (I) is formulated as a lyophilized powder in a manner similar to that described in WO 02 / 059131 by Plamondon et al., which reference is incorporated herein by reference in its entirety. In such embodiments, an aqueous mixture comprising an a-hydroxycarboxylic acid or a β-hydroxycarboxylic acid is lyophilized to form the compound of formula (I).

[0659] In certain embodiments, the lyophilized powder further comprises a free a- hydroxycarboxylic acid or a β-hydroxycarboxylic acid. Preferably, the free a- hydroxycarboxylic acid or β-hydroxycarboxylic acid compound is present in the mixture in a molar ratio ranging from about 0.5: 1 to about 100: 1, more preferably about 5: 1 to about 100: 1, with the compound of formula (I). In various embodiments wherein the a-hydroxycarboxylic acid or β-hydroxycarboxylic acid compound is citric acid, the lyophilized powder comprises a molar ratio of free citric acid to the corresponding borate ranging from about 10: 1 to about 100: 1, about 20: 1 to about 100: 1, or about 40: 1 to about 100: 1.

[0660] In certain embodiments, the lyophilized powder comprises citric acid and the compound of formula (I), substantially free of other components. However, the composition can further comprise one or more other pharmaceutically acceptable excipients, carriers, diluents, fillers, salts, buffers, bulking agents, stabilizers, solubilizers, and other substances well known in the art. The preparation of pharmaceutically acceptable formulations containing these substances is described, for example, in Remington: The Science and Practice of Pharmacy, 20thedition, Editor A. Gennaro, Lippincott Williams & Wilkins, 2000 or more recent edition, and in Strickley, Pharmaceutical Research, 21(2) 201-230 (2004).

[0661] Upon dissolution in an aqueous medium, an equilibrium is established between the borate ester compound of formula (I) and the corresponding free boronic acid compound. In certain embodiments, the equilibrium is rapidly reached, for example within 1-15 minutes after addition of the aqueous medium. The relative concentrations of borate ester, boronic acid, and any intermediates present at equilibrium depend on parameters such as the pH of the solution, the temperature, the nature of the a-hydroxycarboxylic acid or β-hydroxycarboxylic acid, and the ratio of a-hydroxycarboxylic acid or β-hydroxycarboxylic acid to the borate ester compound of formula (I) present in the lyophilized powder.

[0662] In certain embodiments, the pharmaceutical composition comprises the compound of Formula (I), a bulking agent, and a buffer. In certain other embodiments, the pharmaceutical composition comprises the compound of Formula (I), a bulking agent, and a buffer in a lyophilized powder.

[0663] In certain embodiments, the compound of Formula (I) is preformed. In certain other embodiments, the compound of Formula (I) is formed on-site from the corresponding boronic acid of Formula (VIII). In certain other embodiments, the compound (I-1) is preformed. In certain other embodiments, the compound (I-15) is formed on-site from the compound (VIII-15).

[0664] Suitable bulking agents include glycine. In certain embodiments, the bulking agent is present in an amount of about 1% weight by volume (w / v) to about 5% w / v. In certain other embodiments, the bulking agent is present in an amount of about 3% w / v.

[0665] Suitable buffers include sodium citrate, citric acid, and mixtures thereof. In certain embodiments, the buffer is sodium citrate and citric acid.

[0666] In certain embodiments, the buffer is present in a concentration of about 45 mM to about 65 mM. In certain other embodiments, the buffer is present in a concentration of about 50 mM to about 60 mM.

[0667] In certain embodiments, the ratio of buffer to compound of Formula (I) is about 50: 1 to about 10: 1. In certain other embodiments, the ratio of buffer to compound of Formula (I) is about 30: 1 to about 10: 1. In certain other embodiments, the ratio of buffer to compound of Formula (I) is about 20: 1.

[0668] In certain embodiments, the pH of the pharmaceutical composition is between about pH 4.7 and pH 6.1. The pH of the pharmaceutical composition can be adjusted using any suitable inorganic or organic acid.

[0669] In certain embodiments, the pharmaceutical composition comprises the compound of Formula (I), a bulking agent, and a buffer; wherein:

[0670] the a-hydroxy carboxylic acid or the β-hydroxy carboxylic acid is citric acid;

[0671] A is 0;

[0672] R a is isobutyl;

[0673] R a1 is hydrogen, C 1-6 aliphatic, -(CH2) m -CH2-R B or -(CH2) m -CH(R 5a )-OR5b ;

[0674] P is R c -C(O)-;

[0675] R c is -R D ;

[0676] m is 0 or 1 ;

[0677] the bulking agent is glycine; and

[0678] the buffering agent is sodium citrate and citric acid.

[0679] In certain embodiments, the pharmaceutical composition comprises a compound of Formula (I), a bulking agent, and a buffering agent; wherein:

[0680] the compound of Formula (I) is represented by compound (I-1), (I-15), or (I-18);

[0681] the bulking agent is glycine; and

[0682] the buffering agent is sodium citrate and citric acid.

[0683] In certain embodiments, the pharmaceutical composition comprises a compound of Formula (I), a bulking agent, and a buffering agent in a lyophilized powder; wherein:

[0684] the a-hydroxy carboxylic acid or β-hydroxy carboxylic acid is citric acid;

[0685] A is 0;

[0686] R a is isobutyl;

[0687] R a1 is hydrogen, C 1-6 aliphatic, -(CH2) m -CH2-R B or -(CH2) m -CH(R 5a )-OR 5b ;

[0688] P is R c -C(O)-;

[0689] R c is -R D ;

[0690] m is 0 or 1 ;

[0691] the bulking agent is glycine; and

[0692] the buffering agent is sodium citrate and citric acid.

[0693] In certain embodiments, the pharmaceutical composition comprises the compound of Formula (I), an expanding agent, and a buffering agent in a lyophilized powder; wherein:

[0694] The compound of Formula (I) is represented by compound (I-1), (I-15), or (I-18);

[0695] The expanding agent is glycine; and

[0696] The buffering agent is sodium citrate and citric acid.

[0697] In certain embodiments, the pharmaceutical composition comprises compound (I-1) in a lyophilized powder. In certain other embodiments, the pharmaceutical composition comprises compound (I-1), glycine, sodium citrate, and citric acid in a lyophilized powder. In certain other embodiments, the pharmaceutical composition comprises compound (I-15) in a lyophilized powder. In certain other embodiments, the pharmaceutical composition comprises compound (I-15), glycine, sodium citrate, and citric acid in a lyophilized powder.

[0698] In certain embodiments, the present application provides a unit dose pharmaceutical composition comprising a compound of Formula (I-1), an expanding agent, and a buffering agent in a lyophilized powder. In certain embodiments, the unit dose pharmaceutical composition comprises a compound of Formula (I-1), glycine, sodium citrate, and citric acid in a lyophilized powder.

[0699] In certain embodiments, the compound of Formula (I-1) is present in the unit dose pharmaceutical composition in an amount that is about 1 mg to about 10 mg of the equivalent molar weight of the compound of Formula (VIII-1). In certain embodiments, the compound of Formula (I-1) is present in the unit dose pharmaceutical composition in an amount that is about 1 mg to about 5 mg of the equivalent molar weight of the compound of Formula (VIII-1). In certain embodiments, the compound of Formula (I-1) is present in the unit dose pharmaceutical composition in an amount that is about 1.0 mg, about 1.5 mg, about 2.0 mg, about 2.5 mg, about 3.5 mg, about 4.0 mg, about 4.5 mg, or about 5 mg of the equivalent molar weight of the compound of Formula (VIII-1). In certain embodiments, the compound of Formula (I-1) is present in the unit dose pharmaceutical composition in an amount that is about 3.5 mg of the equivalent molar weight of the compound of Formula (VIII-1).

[0700] In certain embodiments, the amount of glycine present in the unit dose pharmaceutical composition is about 0.01 g to about 0.50 g. In certain embodiments, the amount of glycine present in the unit dose pharmaceutical composition is about 0.03 g to about 0.250 g. In certain embodiments, the amount of glycine present in the unit dose pharmaceutical composition is about 0.06 g to about 0.125 g.

[0701] In certain embodiments, sodium citrate and citric acid are present in the unit dose pharmaceutical composition in an amount equivalent to about 0.005 g to about 0.250 g of citrate ion. In certain embodiments, sodium citrate and citric acid are present in the unit dose pharmaceutical composition in an amount equivalent to about 0.025 g to about 0.125 g of citrate ion.

[0702] In certain embodiments, the present application provides a unit dose pharmaceutical composition comprising a compound of Formula (I-15), an expanding agent, and a buffering agent in a lyophilized powder. In certain embodiments, the unit dose pharmaceutical composition comprises a compound of Formula (I-15), glycine, sodium citrate, and citric acid in a lyophilized powder.

[0703] In certain embodiments, the compound of Formula (I-15) is present in the unit dose pharmaceutical composition in an amount equivalent to about 1 mg to about 10 mg of the equivalent molar weight of the compound of Formula (VIII-15). In certain embodiments, the compound of Formula (I-15) is present in the unit dose pharmaceutical composition in an amount equivalent to about 1 mg to about 5 mg of the equivalent molar weight of the compound of Formula (VIII-15). In certain embodiments, the compound of Formula (I-15) is present in the unit dose pharmaceutical composition in an amount equivalent to about 1.0 mg, about 1.5 mg, about 2.0 mg, about 2.5 mg, about 3.5 mg, about 4.0 mg, about 4.5 mg, or about 5 mg of the equivalent molar weight of the compound of Formula (VIII-15). In certain embodiments, the compound of Formula (I-15) is present in the unit dose pharmaceutical composition in an amount equivalent to about 3.5 mg of the equivalent molar weight of the compound of Formula (VIII-15).

[0704] In certain embodiments, the amount of glycine present in the unit dose pharmaceutical composition is about 0.01 g to about 0.50 g. In certain embodiments, the amount of glycine present in the unit dose pharmaceutical composition is about 0.03 g to about 0.250 g. In certain embodiments, the amount of glycine present in the unit dose pharmaceutical composition is about 0.06 g to about 0.125 g.

[0705] In certain embodiments, sodium citrate and citric acid are present in the unit dose pharmaceutical composition in an amount equivalent to about 0.005 g to about 0.250 g of citrate ion. In certain embodiments, sodium citrate and citric acid are present in the unit dose pharmaceutical composition in an amount equivalent to about 0.025 g to about 0.125 g of citrate ion.

[0706] In another aspect, the present application provides a method of preparing a compound of Formula (I) as a lyophilized powder; the method comprising the steps of:

[0707] (d-1) combining the following to form a mixture:

[0708] i. an aqueous solvent mixture;

[0709] ii. a compound of Formula (I);

[0710] iii. an expanding agent; and

[0711] iv. a buffering agent; and

[0712] (d-2) lyophilizing the mixture.

[0713] In certain embodiments, the compound of Formula (I) is formed on-site from the corresponding compound of Formula (VIII). Accordingly, the present application also provides a method of preparing a compound of Formula (I) as a lyophilized powder; the method comprising the steps of:

[0714] (e-1) combining the following to form a mixture:

[0715] i. an aqueous solvent mixture;

[0716] ii. a compound of Formula (VIII);

[0717] iii. an expanding agent; and

[0718] iv. an alpha-hydroxy carboxylic acid or salt thereof, or a beta-hydroxy carboxylic acid or salt thereof, or a combination thereof; and

[0719] (e-2) lyophilizing the mixture.

[0720] In certain embodiments, the aqueous solvent mixture comprises one or more co-solvents in addition to water. In certain embodiments, the co-solvent is miscible with water. In certain other embodiments, the co-solvent is an alcohol, including but not limited to ethanol, t-butanol, and mixtures thereof. In certain other embodiments, the co-solvent is t-butanol.

[0721] In certain embodiments, the aqueous solvent mixture comprises about 1% v / v to about 40% v / v alcohol. In certain other embodiments, the aqueous solvent mixture comprises about 3% v / v to about 10% v / v alcohol. In certain other embodiments, the aqueous solvent mixture comprises about 3% v / v to about 6% v / v alcohol. In certain other embodiments, the solvent mixture comprises about 3% v / v to about 6% t-butanol. In certain other embodiments, the solvent mixture comprises about 5% v / v t-butanol.

[0722] In certain embodiments, a method of preparing Compound (I-1) as a lyophilized powder is provided; the method comprising the steps of:

[0723] (f-1) combining the following to form a mixture:

[0724] i. water;

[0725] ii. Compound (I-1);

[0726] iii. glycine;

[0727] iv. sodium citrate; and

[0728] v. citric acid; and

[0729] (f-2) lyophilizing the mixture.

[0730] In certain embodiments, there is provided a method of preparing compound (I-15) as a lyophilized powder, the method comprising the steps of:

[0731] (g-1) combining the following to form a mixture:

[0732] i. an aqueous solvent mixture comprising water and tert-butanol;

[0733] ii. compound (VIII-15);

[0734] iii. glycine;

[0735] iv. sodium citrate; and

[0736] v. citric acid; and

[0737] (g-2) lyophilizing the mixture.

[0738] In certain other embodiments, for the methods described directly above, the amount of tert-butanol present in the aqueous solvent mixture is from about 3% v / v to about 6% v / v.

[0739] Lyophilization or freeze-drying can be performed using any conventional lyophilizer or freeze-drier. In certain embodiments, lyophilization comprises the steps of: (i) loading the liquid mixture as prepared above, and freezing; (ii) primary drying; (iii) secondary freezing cycle; (iv) drying under vacuum; and (v) secondary drying. The temperature and time for each step depends on the lyophilizer or freeze-drier used.

[0740] In certain embodiments, the resulting lyophilized powder has a residual moisture content of less than about 2%. In certain other embodiments, the resulting lyophilized powder has a residual moisture content of less than about 1%.

[0741] In another aspect, the present application provides a method of preparing a pharmaceutical composition of a compound of Formula (I) as a liquid pharmaceutical dosage form, the method comprising the step of reconstituting a lyophilized powder of a compound of Formula (I) with an aqueous solvent suitable for pharmaceutical administration. Suitable reconstitution solvents include, but are not limited to, water, normal saline, phosphate buffered saline (PBS), and mixtures thereof. In certain embodiments, the reconstitution solvent is water, water for injection, normal saline, and mixtures thereof. In certain other embodiments, the reconstitution solvent is water for injection. After reconstitution, the liquid pharmaceutical dosage form can contain the compound of Formula (I) in a concentration as described herein.

[0742] In certain embodiments, a method of preparing a pharmaceutical composition of Compound (I-1) as a liquid pharmaceutical dosage form is provided, the method comprising the step of reconstituting a lyophilized powder of Compound (I-1) as described herein with an aqueous solvent suitable for pharmaceutical administration. In certain embodiments, a method of preparing a pharmaceutical composition of Compound (I-1) as a liquid pharmaceutical dosage form is provided, the method comprising the step of reconstituting a lyophilized powder of Compound (I-1) as described herein with water for injection or normal saline. In certain embodiments, a method of preparing a pharmaceutical composition of Compound (I-1) as a liquid pharmaceutical dosage form is provided, the method comprising the step of reconstituting a lyophilized powder of Compound (I-1) as described herein with water for injection.

[0743] In certain embodiments, a method of preparing a pharmaceutical composition of Compound (I-15) as a liquid pharmaceutical dosage form is provided, the method comprising the step of reconstituting a lyophilized powder of Compound (I-15) as described herein with an aqueous solvent suitable for pharmaceutical administration. In certain embodiments, a method of preparing a pharmaceutical composition of Compound (I-15) as a liquid pharmaceutical dosage form is provided, the method comprising the step of reconstituting a lyophilized powder of Compound (I-15) as described herein with water for injection or normal saline. In certain embodiments, a method of preparing a pharmaceutical composition of Compound (I-15) as a liquid pharmaceutical dosage form is provided, the method comprising the step of reconstituting a lyophilized powder of Compound (I-15) as described herein with water for injection.

[0744] Upon reconstitution in the reconstitution solvent, an equilibrium is established between the compound of Formula (I) and the corresponding boronic acid of Formula (VIII). Typically, equilibrium is rapidly achieved within about 10-15 minutes after addition of the reconstitution solvent. The relative concentrations of boronate and boronic acid present at equilibrium depend on the pH of the solution, the temperature, and the ratio of the a-hydroxy or β-hydroxy acid compound to the boronic acid compound.

[0745] In another aspect, the present application provides a liquid pharmaceutical composition comprising a compound of Formula (I) and additional excipients described herein. In certain embodiments, the liquid pharmaceutical composition is suitable for parenteral use. In other certain embodiments, the liquid pharmaceutical composition is suitable for oral use.

[0746] In the described embodiments, the liquid pharmaceutical composition comprises the compound of Formula (I), a buffer, and optionally a tonicity adjusting agent.

[0747] In certain embodiments, the ratio of buffer to compound of Formula (I) is about 50: 1 to about 10: 1. In certain other embodiments, the ratio of buffer to compound of Formula (I) is about 30: 1 to about 10: 1. In certain other embodiments, the ratio of buffer to compound of Formula (I) is about 20: 1.

[0748] In certain embodiments, the buffer is present at a concentration of about 45 mM to about 65 mM. In certain other embodiments, the buffer is present at a concentration of about 50 mM to about 60 mM.

[0749] Suitable buffers include sodium citrate, citric acid, and mixtures thereof. In certain embodiments, the buffer is sodium citrate and citric acid.

[0750] Suitable tonicity adjusting agents include, but are not limited to, amino acids, such as arginine, histidine, and glycine; salts, such as sodium chloride, potassium chloride, sodium citrate; propylene glycol; and mixtures thereof. In certain embodiments, the tonicity adjusting agent is propylene glycol. In certain other embodiments, the tonicity adjusting agent is sodium chloride.

[0751] Upon dissolution in the aqueous solvent mixture, an equilibrium is established between the compound of Formula (I) and the corresponding boronic acid of Formula (VIII). Thus, either the compound of Formula (I) or the compound of Formula (VIII) can be used to prepare the liquid pharmaceutical composition. Typically, equilibrium is rapidly reached within about 10-15 minutes after addition of the aqueous solvent mixture. The relative concentrations of boronate ester and boronic acid present at equilibrium depend on the pH of the solution, the temperature, and the ratio of the a-hydroxy or β-hydroxy acid compound to the boronic acid compound. In certain embodiments, an excess of the a-hydroxy or β-hydroxy acid can act as a stabilizer, which drives the equilibrium toward the boronate ester. In certain embodiments, a tonicity adjusting agent can also act as a stabilizer.

[0752] In certain embodiments, the liquid pharmaceutical composition optionally further comprises a preservative.

[0753] In certain embodiments, the liquid pharmaceutical composition comprises the compound of Formula (I), a buffer, and optionally a tonicity adjusting agent; wherein:

[0754] the a-hydroxy carboxylic acid or β-hydroxy carboxylic acid is citric acid;

[0755] A is 0;

[0756] R a is isobutyl;

[0757] R a1 is hydrogen, C1-6 aliphatic, -(CH2) m -CH2-R B or -(CH2) m -CH(R 5a )-OR 5b ;

[0758] P is R c -C(O)-;

[0759] R c is -R D ;

[0760] m is 0 or 1 ;

[0761] the buffering agent is sodium citrate and citric acid; and

[0762] the tonicity adjusting agent, when present, is sodium chloride.

[0763] In certain embodiments, the liquid pharmaceutical composition comprises a compound of Formula (I), a buffering agent, and, optionally, a tonicity adjusting agent; wherein:

[0764] the compound of Formula (I) is represented by compound (I-1), (I-15), or (I-18);

[0765] the buffering agent is sodium citrate and citric acid; and

[0766] the tonicity adjusting agent, when present, is sodium chloride.

[0767] In certain embodiments wherein the a-hydroxy carboxylic acid or β-hydroxy acid is citric acid, the liquid pharmaceutical composition of a compound of Formula (I) comprises a compound of Formula (I), water, citric acid, sodium citrate, and sodium chloride. In certain other embodiments wherein the a-hydroxy carboxylic acid or β-hydroxy carboxylic acid is citric acid, the liquid pharmaceutical composition comprises a compound of Formula (I), water, citric acid, and propylene glycol. In certain other embodiments, the liquid pharmaceutical composition comprises a compound of Formula (I) (wherein the compound of Formula (I) is compound (I-1)), water, citric acid, sodium citrate, and sodium chloride.

[0768] In the described embodiments wherein the a-hydroxy carboxylic acid or β-hydroxy acid is citric acid, the liquid pharmaceutical dosage form of a compound of Formula (I) has a pH value between about pH 3 and about pH 7. In certain of the described embodiments, the pH value is between about pH 4.9 and about pH 6.7. In other certain of the described embodiments, the pH value is between about pH 5.5 and about pH 6.5.

[0769] In certain embodiments, the liquid pharmaceutical composition of the compound of Formula (I) is prepared extemporaneously from a stock vehicle solution and the compound of Formula (VIII). In certain embodiments, the stock vehicle solution comprises water, citric acid, sodium citrate, and propylene glycol. In such embodiments, the resulting solution can be further diluted with the stock vehicle solution or with a sodium chloride solution to produce the liquid pharmaceutical composition of the compound of Formula (I) at the desired concentration.

[0770] In another aspect, the present application provides a unit dose liquid pharmaceutical composition comprising a compound of Formula (I), a buffer, and optionally a tonicity adjusting agent. In certain embodiments, the unit dose liquid pharmaceutical composition comprises a compound of Formula (I), a buffer, and optionally a tonicity adjusting agent, wherein the compound of Formula (I) is compound (I-1). In certain embodiments, the compound of Formula (I) is present in the unit dose liquid pharmaceutical composition at a concentration of about 0.5 mg / ml to about 3 mg / ml of the compound of Formula (VIII). In certain other embodiments, the compound of Formula (I) is present in the unit dose liquid pharmaceutical composition at a concentration of about 1 mg / ml of the compound of Formula (VIII). In certain other embodiments, wherein the compound of Formula (I) is compound (I-1), compound (I-1) is present in the unit dose liquid pharmaceutical composition at a concentration of about 0.5 mg / ml to about 3 mg / ml of the compound of Formula (VIII-1). In certain other embodiments, wherein the compound of Formula (I) is compound (I-1), compound (I-1) is present in the unit dose liquid pharmaceutical composition at a concentration of about 1 mg / ml of the compound of Formula (VIII-1). In certain other embodiments, wherein the compound of Formula (I) is compound (I-15), compound (I-15) is present in the unit dose liquid pharmaceutical composition at a concentration of about 1 mg / ml of the compound of Formula (VIII-15).

[0771] In certain embodiments, the sodium citrate and citric acid are present in the unit dose liquid pharmaceutical composition in an amount equivalent to about 0.005 g to about 0.250 g of citrate ion. In certain embodiments, the sodium citrate and citric acid are present in the unit dose liquid pharmaceutical composition in an amount equivalent to about 0.025 g to about 0.125 g of citrate ion.

[0772] In certain embodiments, the sodium chloride is present in the unit dose liquid pharmaceutical composition in an amount of about 0.0045 g to about 0.09 g. In certain embodiments, the sodium chloride is present in the unit dose liquid pharmaceutical composition in an amount of about 0.01 g to about 0.04 g.

[0773] In certain embodiments of the unit dose liquid pharmaceutical composition, the pharmaceutical composition is stored frozen until use.

[0774] In another aspect, the present application provides a method of preparing a compound of Formula (I) as a unit dose liquid pharmaceutical composition, the method comprising the steps of:

[0775] (h-1) dissolving a buffer in an aqueous solvent;

[0776] (h-2) dissolving a compound of Formula (I) or a crystalline form thereof in the mixture resulting from step (h-1);

[0777] (h-3) dissolving a tonicity adjusting agent in the mixture resulting from step (h-2);

[0778] (h-4) adding additional aqueous solvent to the desired batch volume; and

[0779] (h-5) filling vials with an amount of the mixture resulting from step (h-4).

[0780] In certain embodiments, the vials are capped after step (h-5). In certain other embodiments, nitrogen gas is bubbled through the mixture prior to step (h-5). In certain other embodiments, after step (h-5), the liquid in the vials can be covered with nitrogen gas prior to capping.

[0781] In certain embodiments, the compound of Formula (I) is formed on-site from a compound of Formula (VIII). In such embodiments, in step (h-2), a compound of Formula (VIII) or a crystalline form thereof is added to the mixture. In certain embodiments, an alpha-hydroxy acid or a beta-hydroxy acid is added in step (h-2). In certain other embodiments, the alpha-hydroxy acid or the beta-hydroxy acid is present as the buffer in step (h-1).

[0782] The pharmaceutical compositions of the present application are preferably formulated so as to be administered to a patient suffering from a proteasome-mediated disorder or at risk of developing or experiencing a recurrence of a proteasome-mediated disorder. As used herein, the term "patient" means an animal, preferably a mammal, more preferably a human. Preferred pharmaceutical compositions of the present application are those that are formulated for oral, intravenous, or subcutaneous administration. However, any of the above-mentioned dosage forms containing a therapeutically effective amount of a compound of the present application are well within the scope of routine experimentation and are therefore well within the scope of the present application. In certain embodiments, the pharmaceutical compositions of the present application can further comprise other therapeutic agents. In certain embodiments, the other therapeutic agents are therapeutic agents that are typically administered to a patient suffering from the disease or condition being treated.

[0783] "Therapeutically effective amount" means an amount sufficient to effect a detectable reduction in the severity of proteasome activity or a proteasome-mediated disorder. The amount of proteasome inhibitor needed will depend on the effectiveness of the inhibitor for a given cell type and the duration of treatment required for the disorder. It will also be appreciated that the specific dose and treatment regimen for any particular patient will depend on a variety of factors, including the activity of the specific compound employed, the age, body weight, general health, sex, and diet of the patient, the time of administration, rate of excretion, drug combination, the judgment of the treating physician, and the severity of the particular disease being treated. The amount of additional therapeutic agent present in the compositions of this application will generally not exceed the amounts normally administered in compositions comprising that therapeutic agent as the sole active agent. Preferably, the amount of additional therapeutic agent is in the range of about 50% to about 100% of the amount normally present in compositions comprising that agent as the sole therapeutically active agent.

[0784] In another aspect, the present application provides methods of treating a patient suffering from a proteasome-mediated disorder or at risk of developing or experiencing a recurrence of a proteasome-mediated disorder. As used herein, the term "proteasome-mediated disorder" includes any disorder, disease, or condition caused by or characterized by an increase in proteasome expression or activity or requiring proteasome activity. The term "proteasome-mediated disorder" also includes any disorder, disease, or condition in which inhibition of proteasome activity is beneficial.

[0785] For example, the compounds and pharmaceutical compositions of the present application are useful in the treatment of disorders mediated by proteins regulated by proteasome activity, such as NFKB, p27 Kip , p21 WAF / CIP1 , p53. Related disorders include inflammatory disorders (e.g., rheumatoid arthritis, inflammatory bowel disease, asthma, chronic obstructive pulmonary disease (COPD), osteoarthritis, skin disorders (e.g., atopic dermatitis, psoriasis)), vascular proliferative disorders (e.g., atherosclerosis, restenosis), proliferative ocular disorders (e.g., diabetic retinopathy), benign proliferative disorders (e.g., hemangioma), autoimmune diseases (e.g., multiple sclerosis, tissue and organ rejection), and inflammation associated with infection (e.g., immune response), neurodegenerative disorders (e.g., Alzheimer's disease, Parkinson's disease, motor neuron disease, neuropathic pain, trinucleotide repeat disorders, astrocytomas, and neurodegeneration due to alcoholic liver disease), ischemic injury (e.g., stroke), and cachexia (e.g., accelerated muscle proteolysis accompanying various physiological and pathological conditions such as nerve injury, fasting, fever, acidosis, HIV infection, cancer cachexia, and certain endocrine diseases).

[0786] The compounds and pharmaceutical compositions of the present invention are particularly useful for the treatment of cancer. As used herein, the term "cancer" refers to a cellular disorder characterized by uncontrolled or abnormal regulation of cell proliferation, decreased cell differentiation, inappropriate ability of cells to invade surrounding tissues, and / or ability to establish new growths at ectopic sites. The term "cancer" includes, but is not limited to, solid tumors and blood-borne tumors. The term "cancer" encompasses diseases of the skin, tissues, organs, bone, cartilage, blood, and blood vessels. The term "cancer" further encompasses primary and metastatic cancers.

[0787] Non-limiting examples of solid tumors that can be treated with the disclosed proteasome inhibitors or pharmaceutical compositions include pancreatic cancer; bladder cancer; colorectal cancer; breast cancer, including metastatic breast cancer; prostate cancer, including androgen-dependent and androgen-independent prostate cancer; kidney cancer, including, for example, metastatic renal cell carcinoma; hepatocellular carcinoma; lung cancer, including, for example, non-small cell lung cancer (NSCLC), bronchioloalveolar carcinoma (BAC), and lung adenocarcinoma; ovarian cancer, including, for example, progressive epithelial or primary peritoneal cancer; cervical cancer; gastric cancer; esophageal cancer; head and neck cancer, including, for example, squamous cell carcinoma of the head and neck; melanoma; neuroendocrine cancer, including metastatic neuroendocrine tumor; brain tumors, including, for example, glioma, anaplastic oligodendroglioma, adult

[0788] Non-limiting examples of hematologic malignancies that can be treated with the disclosed proteasome inhibitors or pharmaceutical compositions include acute myelogenous leukemia (AML); chronic myelogenous leukemia (CML), including accelerated CML and CML blast phase (CML-BP); acute lymphoblastic leukemia (ALL); chronic lymphocytic leukemia (CLL); Hodgkin's disease (HD); non-Hodgkin's lymphoma (NHL), including follicular lymphoma and mantle cell lymphoma; B-cell lymphoma; T-cell lymphoma; multiple myeloma (MM); Waldenstrom's macroglobulinemia; myelodysplastic syndrome (MDS), including refractory anemia (RA), refractory anemia with ringed siderblasts (RARS), refractory anemia with excess blasts (RAEB), and RAEB in transformation (RAEB-T); and myeloproliferative syndromes.

[0789] In certain embodiments, the compounds or pharmaceutical compositions of the present invention are used to treat a patient having cancer or at risk of developing or experiencing a recurrence of cancer selected from the group consisting of multiple myeloma and mantle cell lymphoma.

[0790] In certain embodiments, a proteasome inhibitor or pharmaceutical composition of the present application is administered in conjunction with other therapeutic agents. The other therapeutic agents can also inhibit proteasomes, or can operate by a different mechanism. In certain embodiments, the other therapeutic agents are therapeutic agents that are typically administered to a patient having the disease or condition being treated. The proteasome inhibitors of the present application can be administered with the other therapeutic agents in a single dosage form or in separate dosage forms. When administered in separate dosage forms, the other therapeutic agents can be administered prior to, simultaneously with, or following administration of the proteasome inhibitors of the present application.

[0791] In certain embodiments, a proteasome inhibitor of Formula (I) or a pharmaceutical composition of a compound of Formula (I) is administered in conjunction with an anti-cancer agent. As used herein, the term "anti-cancer agent" refers to any agent that is administered to an individual having a cancer for the purpose of treating the cancer.

[0792] Non-limiting examples of DNA damaging chemotherapeutic agents include topoisomerase I inhibitors (e.g., irinotecan, topotecan, camptothecin and analogs or metabolites thereof, and doxorubicin); topoisomerase II inhibitors (e.g., etoposide, teniposide, and daunorubicin); alkylating agents (e.g., melphalan, chlorambucil, busulfan, thiotepa, ifosfamide, carmustine, lomustine, semustine, streptozocin, decarbazine, methotrexate, mitomycin C, and cyclophosphamide); DNA intercalators (e.g., cisplatin, oxaliplatin, and carboplatin); DNA intercalators and free radical generators (e.g., bleomycin); and nucleoside mimetics (e.g., 5-fluorouracil, capecitibine, gemcitabine, fludarabine, cytarabine, mercaptopurine, thioguanine, pentostatin, and hydroxyurea).

[0793] Chemotherapeutic agents that interfere with cell replication include: paclitaxel, docetaxel, and related analogs; vincristine, vinblastin, and related analogs; thalidomide, lenalidomide, and related analogs (e.g., CC-5013 and CC-4047); protein tyrosine kinase inhibitors (e.g., imatinib mesylate and gefitinib); proteasome inhibitors (e.g., bortezomib); NF-κB inhibitors, including IκB kinase inhibitors; antibodies that bind to proteins that are overexpressed in cancer and thereby down-regulate cell replication (e.g., trastuzumab, rituximab, cetuximab, and bevacizumab); and other inhibitors of proteins or enzymes that are known to be up-regulated, overexpressed, or activated in cancer, the inhibition of which down-regulates cell replication.

[0794] In order that the present application can be more fully understood, the following preparation and testing examples are set forth. These examples illustrate methods of making or testing particular compounds and are not to be construed as limiting the scope of the application in any way.

[0795] Example

[0796] Abbreviations

[0797] DCM dichloromethane

[0798] DIPEA N,N'-diisopropylethylamine

[0799] DMF N,N'-dimethylformamide

[0800] EDCI N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride

[0801] EtOAc ethyl acetate

[0802] h hour(s)

[0803] HPLC high performance liquid chromatography

[0804] MIBK methyl isobutyl ketone

[0805] PES polyethersulfone

[0806] TBTU O-benzotriazol-1-yl-N,N,N',N'-tetramethyluronium tetrafluoroborate

[0807] TFA trifluoroacetic acid

[0808] THF tetrahydrofuran

[0809] HOBt 1-hydroxybenzotriazole

[0810] LCMS liquid chromatography mass spectrometry

[0811] min minutes

[0812] General Methods

[0813] 1 H NMR: Spectra were obtained at ambient temperature via a JOEL ECX-400 NMR spectrometer operating at 400 MHz. The resulting FID was transferred to a PC and processed using NUTS NMR processing software by Acorn NMR Inc. Chemical shifts were referenced to the DMSO solvent (2.50 ppm). A solvent blank was prepared by adding approximately 0.75 mL of DMSO-d6 to an NMR tube. The solvent blank was obtained at ambient temperature and was used as a reference for all samples. The solvent blank was obtained at ambient temperature and was used as a reference for all samples. 1 After the H spectrum, the sample was added and allowed to fully dissolve.

[0814] Mass Spectrometry: Mass spectrometric studies were performed via a Thermo-Finnigan LCQ Deca-XP ion trap mass spectrometer. The electrospray ion source was used in both positive and negative modes at a high voltage of 5 kv, a sheath gas flow rate of 35 arb, a capillary temperature of 275 °C, a capillary voltage of 9 V, and a tube lens compensation voltage of 35 V. The analyte was dissolved in acetonitrile to yield a 0.5 mg / ml solution. The LC-mass spectrometry flow analysis was performed using an Agilent 1100 HPLC system. The pump flow rate was 1.0 milliliter per minute. Ten μl of each sample solution was injected from an autosampler into a T-junction. Approximately 2% of the solution was input from the T-junction into the mass spectrometer.

[0815] X-ray powder diffraction (XRPD): X-ray powder diffraction patterns were obtained via either:

[0816] i) a Bruker AXS D8 Advance diffractometer. Data were collected in continuous scan mode using a step size of 0.05° 2Θ and a step time of 2 seconds over a 2.9° to 29.6° 2Θ angular range. The sample was operated at ambient conditions and prepared as a flat plate sample using the as-received, ungrounded powder; or

[0817] ii) PANalytical X'Pert Pro diffractometer. Each sample was analyzed using Cu radiation generated with Optix long fine focus source. An elliptically tilted multilayer mirror was used to focus the Cu Ka X-rays of the source through the sample and onto the detector. The sample was clamped between 3-micron thick films, transmission geometry was analyzed, and rotated to optimize orientation statistics. A beam stop was used to minimize background from air scatter. No helium and anti-diffraction extension were used. Soller slits were used for the incident and diffracted beams to minimize axial divergence. A scan position with minimized axial divergence was used to collect the diffraction pattern. A scan position sensitive detector (X'Celerator) positioned 240 mm from the sample was used to collect the diffraction pattern. Prior to analysis, a silicon sample (NIST Standard Reference Material 640c) was analyzed to verify the position of the silicon 111 peak.

[0818] Differential Scanning Calorimetry (DSC): Differential Scanning Calorimetry (DSC) data was collected via either:

[0819] i) TA Instruments Q100 Differential Scanning Calorimeter equipped with a 50 position auto-sampler. The energy and temperature calibration standards were indium. The sample was heated at a rate of 10 °C per minute between 25 °C and 300 °C. Purified nitrogen was flowed at 50 mL per minute over the sample during the scan. Between 1 mg and 3 mg of sample was analyzed. All samples were crimped in an aluminum pan which was sealed in a hermetic fashion with a pinhole to relieve pressure built up by solvent vapors; or

[0820] ii) TA Instruments Differential Scanning Calorimeter 2920. The sample was placed in an aluminum DSC pan and the weight was recorded accurately. The open pan was covered with a lid and then crimped. The sample chamber was equilibrated at 25 °C and heated at a rate of 10 °C / minute under nitrogen purge. Indium metal was used as a calibration standard.

[0821] Thermogravimetric Analysis (TGA): Thermogravimetric Analysis (TGA) data was collected via a TA Instruments Q500 Thermogravimetric Analyzer, calibrated with Nickel / Alumel and operated at a scan rate of 10 °C per minute. Purified nitrogen was flowed at 60 mL per minute over the sample during the measurement. Typically, 5 mg to 15 mg of sample was loaded onto a pre-tared platinum crucible.

[0822] Example 1: Synthesis of 4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5- dichlorobenzamido)acetylamino)-3-methylbutyl)-6-oxo-1,3,2-dioxaborolane-4- carboxylic acid (1-1)

[0823]

[0824] Step 1: 2,5-[(Dichlorobenzoyl)amino]acetic acid

[0825] To a mixture of NaOH (12 g, 300 mmol) and glycine (18 g, 239 mmol) in water (120 mL) was added a solution of 2,5-dichlorobenzoyl chloride (10 g, 48 mmol) in THF (15 mL) dropwise over 45 min, keeping the internal temperature below about 25 °C. After 1 h, the mixture was acidified with 2.0 M HC1 (125 mL), keeping the internal temperature below about 5 °C. The resulting precipitate was collected by vacuum filtration. The crude product was recrystallized from water to give 2,5-[(dichlorobenzoyl)amino]acetic acid as a white crystalline solid (6.1 g, 52%). mp 173.3 °C. 1 H NMR (300 MHz, DMSO-d6, δ): 12.72 (bs, 1H), 8.89 (t, J = 6.0 Hz, 1H), 7.54 (m, 2H), 7.48 (m, 1H), 3.93 (d, J = 6.0 Hz). 13 C NMR (75 MHz, DMSO-d6, δ): 41.6, 129.3, 129.6, 131.4, 132.2, 138.2, 171.4, 165.9. MS (m / z): [M+H] calcd for C9H8Cl2NO3 248.0; found 248.0; [M+Na] calcd for C9H7Cl2NNaO3 270.0; found 270.2.

[0826] 2,5-[(Dichlorobenzoyl)amino]acetic acid was also prepared via the following procedure: To a mixture of glycine (21.5 g, 286 mmol) in water (437 mL) was added 2.0 M NaOH (130 mL) and the resulting solution was cooled to 0 °C. A solution of 2,5-dichlorobenzoyl chloride (50.0 g, 239 mmol) in THF (75 mL) was added dropwise at such a rate that the internal temperature was maintained at 0 ± 1 °C. During the addition, the pH was controlled at 11.0 ± 0.2 by titration with 2.0 M NaOH using a pH controller. After the addition was complete, the mixture was stirred at 0 ± 1 °C for an additional 2 h. The mixture was then acidified with 2.0 M HC1 (176 mL) to a final pH of 2.5. The resulting precipitate was collected by filtration, washed with cold water (125 mL), and dried in a vacuum oven at 45 °C to give 2,5-[(dichlorobenzoyl)amino]acetic acid as a white solid (57.6 g, 97.3%).

[0827] Step 2: 2,5-Dichloro-N-[2-({(1R)-3-methyl-1-[(3aS,4S,6S,7aR)-3a,5,5- trimethylhexahydro-4,6-methano-1,3,2-benzodioxol-2-yl]butyl}amino)-2- oxoethyl]benzamide

[0828] To a solution of 2,5-[(dichlorobenzamido]acetic acid (6.10 g, 24.6 mmol) and TBTU (8.34 g, 26.0 mmol) in DMF (40 mL) at an internal temperature below about 5 °C was added (1R)-3-methyl-1-[(3aS,4S,6S,7aR)-3a,5,5-trimethylhexahydro-4,6-methano-1,3,2- benzodioxol-2-yl]butan-1-amine·TFA (9.35 g, 24.7 mmol). DIPEA (13 mL, 75 mmol) was then added dropwise over 2 h, maintaining an internal temperature below about 5 °C. After 40 min, the mixture was diluted with EtOAc (90 mL), washed with 5% NaCl (150 mL), washed twice with 10% NaCl (2 x 40 mL), washed once with 2% K2CO3 (1 x 40 mL), washed once with 1% H3PO4 (1 x 40 mL), and washed once with 10% NaCl (1 x 40 mL). The resulting organic layer was concentrated to a thick oil, diluted with heptane (40 mL), and evaporated to yield 2,5-dichloro-N-[2-({(1R)-3-methyl-1-[(3aS,4S,6S,7aR)-3a,5,5-trimethylhexahydro-4,6-methano-1,3,2-benzodioxol-2-yl]butyl}amino)-2-oxoethyl]benzamide as a white solid, which was used without purification in the next step.

[0829] Step 3: N,N',N”-{Cyclotriboroxane-2,4,6-triyl tris[[(1R)-3-methylbutane-1,1- diyl]imino(2-oxoethane-2,1-diyl)]}tris(2,5-dichlorobenzamide)

[0830] To a solution of 2,5-dichloro-N-[2-({(1R)-3-methyl-1-[(3aS,4S,6S,7aR)- 3a,5,5-trimethylhexahydro-4,6-methano-1,3,2-benzodioxol-2-yl]butyl}amino)-2- oxoethyl]benzamide (12.2 g, 24.6 mmol) in methanol / hexanes (1:1) (250 mL) was added 1 N HC1 (30 mL, 30 mmol) and (2-methylpropyl)boronic acid (6.5 g, 64 mmol). The reaction mixture was stirred overnight. The phases were separated and the methanol layer was washed twice with additional hexanes (2 x 55 mL). The resulting organic layer was concentrated to about 10 mL and partitioned between 2.0 M NaOH (30 mL) and DCM (25 mL). The DCM layer was washed once with additional 2.0 M NaOH (5 mL). The basic aqueous layers were then combined, washed with DCM (2 x 25 mL), and acidified with 1 M HC1 (60 mL). The resulting mixture was diluted with DCM (40 mL), the layers were separated, and the resulting aqueous layer was washed three times with DCM (3 x 10 mL). The combined DCM extracts were dried over MgS04(25 g) and evaporated to a thick oil. The product was precipitated with hexanes (50 mL) and collected by filtration to yield N,N',N''-{boroxin-2,4,6-trisyltris[[(1R)-3-methylbutane-1,1- diyl]imino(2-oxoethane-2,1-diyl)]}tris(2,5-dichlorobenzamide) as a white solid (6.6 g, 74%). 1 HNMR (300 MHz, DMSO-d6, δ): 8.93 (t, J = 6.0 Hz, 1H), 8.68 (bs, 1H), 7.63 (m, 1H), 7.52 (m, 2H), 4.00 (d, J = 6.0 Hz, 2H), 2.62 (m, 1H), 1.59 (m, 1H), 1.33 (m, 1H), 1.24 (m, 1H), 0.81 (d, J = 5.9 Hz, 6H). 13 C NMR (125 MHz, DMSO-d6, δ): 23.2, 25.8, 40.1, 40.7, 43.0, 129.0, 130.0, 131.0, 137.5, 165.0, 172.5. MS (m / z) in CH3CN: [M+H] C 42 H 52 B3Cl6N6O9Calcd 1027.2; Found 1027.3; [M+Na] C 42 H 51 B3Cl6N6NaO9Calcd 1049.2; Found 1049.5.

[0831] Step 4: 4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetylamino)-3- methylbutyl)-6-oxo-1,3,2-dioxaborolane-4-carboxylic acid (1-1)

[0832] Form 1: To a solution of citric acid (2.75 g, 14.3 mmol) in EtOAC (85 mL) at an internal temperature of about 74 °C was added N,N',N''-{boroxin-2,4,6-trisyl tris[[(1R)-3- methylbutane-1,1-diyl]imino(2-oxoethane-2,1-diyl)]}tris(2,5-dichlorobenzamide) (5.00 g, 4.87 mmol) as a solid. The solution was allowed to cool freely until the internal temperature was about 25 °C and the mixture was stirred overnight. The resulting precipitate was collected by filtration to yield 2,2'-{2-[(1R)-1-({[(2,5-dichlorobenzoyl)amino]acetyl}amino)-3- methylbutyl]-5-oxo-1,3,2-dioxaborolan-4,4-diyl}diacetic acid Form 1 (6.65 g, 88%) as a crystalline solid. 1 H NMR (500 MHz, DMSO-d6, δ 110 °C): 10.08 (s, 1H), 8.69 (s, 1H), 7.61 (s, 1H), 7.52 (d, J = 1.3 Hz, 2H), 4.26 (d, J = 5.5 Hz, 2H), 2.70 (q, J = 14.5 Hz, 4H), 2.70 (bs, 1H), 1.72 (sept, J = 6.5 Hz, 1H), 1.42 (ddd, J = 5.2 Hz, J = 8.6 Hz, J = 13.9 Hz, 1H), 1.28 (ddd, J = 5.3, J = 9.4 Hz, J = 14.3 Hz, 1H), 0.91 (dd, J = 3.3 Hz, J = 6.6 Hz, 6H). MS (m / z): [M+Na] C 20 H 23 Calculated for BCl2N2NaO9 539.1; found 539.1.

[0833] XRPD data for I-1 Form 1 is shown in Figure 1 and Table 1.

[0834] 2 theta angle (°) Intensity % 6.441 100 8.304 29.5 10.35 19 11.619 5.1 12.695 13.6 15.077 28.2 16.352 28.7 17.504 16.3 18.231 6 19.086 21.4 20.405 11.7 21.231 7.6 21.916 7.6 25.371 15.2 27.588 6.2

[0835] Table 1: XRPD data for I-1 Form 1

[0836] Differential scanning calorimetry (DSC) data for I-1 Form 1 is shown in Figure 2The plot is characteristic of an endothermic transition with an onset temperature of 191.8 °C and a melting temperature of 198.8 °C. A second endothermic transition corresponding to decomposition has an onset temperature of 225 °C. These temperatures are accurate to ± 5 °C.

[0837] Thermogravimetric analysis (TGA) data for Form I-1 is shown in Figure 2 The plot depicts the percent weight loss of the sample versus temperature at a rate of change of about 10 °C / min. The weight loss represents a loss of about 0.72% of the sample weight when the temperature is changed from 50 °C to 200 °C. These temperatures are accurate to ± 5 °C.

[0838] Form 2: To a solution of citric acid (10.1 g, 52.6 mmol) in EtOAC (300 mL) at an internal temperature of about 74 °C was added a solution of N,N',N"-{boroxin-2,4,6-trisyl tris[[(1R)-3-methylbutane-1,1-diyl]imino(2- oxoethane-2,1-diyl)]} tri(2,5-dichlorobenzamide) (20.0 g, 19.5 mmol) in EtOAC (60 mL). The solution was slowly cooled (at a rate of about 0.33 °C / min) until the internal temperature was about 60 °C and the mixture was stirred for 3 hours. The resulting slurry was slowly cooled (at a rate of about 0.12 °C / min) until the internal temperature was about 25 °C and the mixture was stirred overnight. The resulting precipitate was collected by filtration to yield 4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5- dichlorobenzamido)acetamido)-3-methylbutyl)-6-oxo-1,3,2-dioxaborolane-4-carboxylic acid Form 2 (26.7 g, 98%) as a crystalline solid. 1 H NMR (500 MHz, DMSO-d6, δ 110 °C): 10.08 (s, 1H), 8.69 (s, 1H), 7.61 (s, 1H), 7.52 (d, J = 1.3 Hz, 2H), 4.26 (d, J = 5.5 Hz, 2H), 2.70 (q, J = 14.5 Hz, 4H), 2.70 (bs, 1H), 1.72 (sept, J = 6.5 Hz, 1H), 1.42 (ddd, J = 5.2 Hz, J = 8.6 Hz, J = 13.9 Hz, 1H), 1.28 (ddd, J = 5.3, J = 9.4 Hz, J = 14.3 Hz, 1H), 0.91 (dd, J = 3.3 Hz, J = 6.6 Hz, 6H). 13C NMR (100 MHz, DMSO-d6, δ 100 °C): 21.65, 23.34, 25.09, 38.39, 38.98, 42.07, 76.25, 128.97, 129.14, 130.94, 131.48, 131.73, 137.05, 165.44, 170.23, 175.74, 177.43. MS (m / z): [M+Na] C 20 H 23 BC12N2Na09Calcd 539.1 ; Found 539.1.

[0839] 4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetylamino)-3- methylbutyl)-6-oxo-1,3,2-dioxaborolane-4-carboxylic acid Form 2 was also prepared by adding a solution of citric acid (21 g, 0.11 mmol) in THF (80 mL) to a solution of N,N',N''-{dioxan-2,4,6-triyl tri[[(1 R)-3-methylbutane-1,1 - diyl]imino(2-oxoethane-2,1 -diyl)]} tri(2,5-dichlorobenzamide) (40 g, 0.11 mmol) in THF (80 mL) at 60 °C. The solution was then seeded with Form 2 crystals (400 mg). After stirring at 60 °C for 30 minutes, EtOAC (400 mL) was added over 9 hours. After the addition of EtOAc was complete, the temperature was reduced to 20 °C over 5 hours. The resulting suspension was filtered to collect 4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetylamino)-3-methylbutyl)-6-oxo-1,3,2-dioxaborolane-4-carboxylic acid Form 2 (40 g, 70%) as a crystalline solid.

[0840] 4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetylamino)-3- methylbutyl)-6-oxo-1,3,2-dioxaborolane-4-carboxylic acid Form 2 was also prepared in the same general manner using the conditions described in Table 2.

[0841] Solvent Onset temperature Inoculation temperature Isolation yield of Form 2 of I-1 Acetonitrile 80℃ Uninoculated 77％ MIBK 80℃ Uninoculated 80％ 2-methyltetrahydrofuran 80℃ 60℃ 72％

[0842] Table 2: Additional conditions for preparing Form 2 of 1-1

[0843] 4-(R,S)-(carboxymethyl)-2-((R)-l-(2-(2,5-dichlorobenzamido)acetylamino)-3- methylbutyl)-6-oxo-l,3,2-dioxaborolane-4-carboxylic acid Form 2 was also prepared by dissolving in acetone followed by the addition of EtOAc as an antisolvent.

[0844] XRPD data for I-l Form 2 is shown in Figure 3 and Table 3.

[0845] 2 theta angle (°) Intensity % 5.817 100 7.614 93.4 11.575 71.1 11.896 67.1 12.571 24.3 14.43 32.2 16.689 65.8 17.362 17.8 18.232 53.9 19.596 77.6 19.959 63.8 20.376 36.2 20.998 32.2 21.5 40.1 21.764 43.4 22.407 77.6 23.12 33.6 23.901 26.3 24.402 20.4 24.882 19.7 25.764 19.1 26.464 39.5 27.347 21.7 27.65 17.1 27.979 16.4 29.41 20.4

[0846] Table 3: XRPD data for I-l Form 2

[0847] Differential scanning calorimetry (DSC) data for I-l Form 2 is shown in Figure 4 The plot is characterized by an endothermic transition with an onset temperature of 206.5 °C and a melting temperature of 219.9 °C. A second endothermic transition corresponding to decomposition has an onset temperature of 225 °C. These temperatures are in error by ± 5 °C.

[0848] Thermogravimetric analysis (TGA) data for I-l Form 2 is shown in Figure 4 The plot depicts the percent weight loss of the sample versus temperature at a rate of change of temperature of about 10 °C / minute. The weight loss represents a loss of about 1.1% of the weight of the sample as the temperature is changed from 50 °C to 200 °C. These temperatures are in error by ± 5 °C.

[0849] Example 1A: Alternative synthesis of 4-(R,S)-(carboxymethyl)-2-((R)-l-(2-(2,5- dichlorobenzamido)acetylamino)-3-methylbutyl)-6-oxo-l,3,2-dioxaborolane-4- carboxylic acid (I-l) Form 2

[0850] A 50 L glass reactor, equipped with a mechanical stirrer, dropping funnel, temperature indicator, and heating / cooling control unit, was charged with 1.2 micron filtered EtOAC (18.9 kg) and anhydrous citric acid (0.561 kg, 2.9 mol) under nitrogen. The mixture was heated to 71 °C and a solution was generated. N,N',N"-{cycloboronane-2,4,6-trisyl tris[[(1R)-3-methylbutane-1,1-diyl]imino(2- oxoethane-2,1-diyl)]}tris(2,5-dichlorobenzamide) (1.109 kg, 3.1 mol) dissolved in EtOAC (4.0 kg) was clarified using an in-line filter (1.2 micron) and the solution was added to the reaction mixture over 20 minutes with stirring (193 rpm) while maintaining a temperature of 73-75 °C. The stirring was reduced to 96 rpm and the mixture was cooled as follows: (1) the mixture was held at 73-75 °C for 25 minutes; (2) the mixture was gradually cooled to 40 °C at a rate of about 5 °C / 30 min; (3) the mixture was allowed to cool freely to ambient temperature with stirring overnight. The product was then isolated by filtration, washed on the filter with 1.2 micron filtered EtOAC (2 x 1.2 kg), and dried under vacuum at 40-41 °C overnight (22 hours) to give 1.458 kg (92%) of the title compound. 1 H NMR (400 MHz, DMSO-d6, δ): 12.13 (s, 2H), 10.69 (s, 1H), 9.11 (t, J = 5.6 Hz, 1H), 7.66 (t, J = 1.2 Hz, 1H), 7.56 (d, J = 1.2 Hz, 2H), 4.27 (bs, 2H), 2.9-2.55 (m, 5H), 1.67 (bs, 1H), 1.4-1.15 (bs, 2H), 0.86 (d, J = 6.4 Hz, 6H).

[0851] XRPD data for Compound (I-1) Form 2 is shown in Figure 7 and Table 6.

[0852]

[0853]

[0854] Table 6

[0855] Differential scanning calorimetry (DSC) data for Compound (I-1) Form 2 is shown in Figure 8 The plot is characterized by two endothermic transitions; the first has a melting temperature of about 231.3 °C and the second has a melting temperature of about 239.9 °C. These temperatures are in error by ± 5 °C.

[0856] Example 2: Synthesis of 2,5-dichloro-N-(2-{[(1R)-3-methyl-1-(4-oxo-1,3,2- dioxaborolan-2-yl)butyl]amino}-2-oxoethyl)benzamide (1-2)

[0857] To a solution of glycolic acid (0.041 g, 0.54 mmol) in EtOAC (2.0 mL) at an internal temperature of about 60 °C was added a solution of N,N',N”-{boroxin-2,4,6- triyl tris[[(1R)-3-methylbutane-1,1-diyl]imino(2-oxoethane-2,1-diyl)]}tris(2,5- dichlorobenzamide) (0.199 g, 0.19 mmol) in EtOAC (1.0 mL). The solution was allowed to cool freely until the internal temperature was about 25 °C and the solvent was removed by evaporation to yield 2,5-dichloro-N-(2-{[(1R)-3-methyl-1-(4-oxo-1,3,2- dioxaborolan-2-yl)butyl]amino}-2-oxoethyl)benzamide (0.215 g, 95%) as a white solid. MS (m / z): [M+Et3N+H] C 22 H 35 Calculated for BCl2N3O5 502.2; found 502.0. MS (m / z): [M-H] C 16 H 18 Calculated for BCl2N2O5 399.1; found 399.0.

[0858] Example 3: Synthesis of {(4S)-2-[(1R)-1-({[(2,5-dichlorobenzoyl)amino]-acetyl}amino)-3- methylbutyl]-5-oxo-1,3,2-dioxaborolan-4-yl}acetic acid (1-3)

[0859] To a solution of L-malic acid (0.0958 g, 0.714 mmol) in EtOAC (2.0 mL) at an internal temperature of about 60 °C was added a solution of N,N',N”-{boroxin-2,4,6- triyl tris[[(1R)-3-methylbutane-1,1-diyl]imino(2-oxoethane-2,1-diyl)]}tris(2,5- dichlorobenzamide) (0.239 g, 0.233 mmol) in EtOAC (1.0 mL). The solution was allowed to cool freely until the internal temperature was about 25 °C and the solvent was removed by evaporation to yield {(4S)-2-[(1R)-1-({[(2,5-dichlorobenzoyl)amino]- acetyl}amino)-3-methylbutyl]-5-oxo-1,3,2-dioxaborolan-4-yl}acetic acid (0.307 g, 96%) as a white solid. MS (m / z): [M+Et3N+H] C24 H 37 BC12N3O7Calcd 560.1 ; Found 560.1. MS (m / z) in CH3CN: [M-H] C 18 H 20 BC12N2O7Calcd 457.1 ; Found 457.1.

[0860] Example 4: Synthesis of 2,5-dichloro-N-[2-({(1R)-1-[(4S)-4-cyclohexyl-5-oxo-1,3,2- dioxaborolan-2-yl]-3-methylbutyl}amino)-2-oxoethyl]benzamide (1-4)

[0861] To a solution of (S)-hexahydromandelic acid (0.0881 g, 0.557 mmol) in EtOAC (2.0 mL) at an internal temperature of about 60 °C was added a solution of N,N',N”-{cycloboroxane-2,4,6- triyl tris[[(1R)-3-methylbutane-1,1-diyl]imino(2-oxoethane-2,1-diyl)]} tri(2,5-dichlorobenzamide) (0.200 g, 0.195 mmol) in EtOAC (1.0 mL). The solution was allowed to cool freely until the internal temperature was about 25 °C and the solvent was removed by evaporation to yield 2,5-dichloro-N-[2-({(1R)-1-[(4S)-4-cyclohexyl-5-oxo-1,3,2-dioxaborolan-2-yl]-3- methylbutyl}amino)-2-oxoethyl]benzamide (0.251 g, 93%) as a white solid. MS (m / z) in CH3CN: [M+Et3N+H] C 28 H 45 BC12N3O5Calcd 584.3; Found 584.1. MS (m / z) in CH3CN: [M-H] C 22 H 28 BC12N2O5Calcd 481.1 ; Found 481.1.

[0862] Example 5: Synthesis of 2,5-dichloro-N-(2-{[(1R)-1-(4,4-dimethyl-5-oxo-1,3,2-dioxaborolan-2- yl)-3-methylbutyl]amino}-2-oxoethyl)benzamide (1-5)

[0863] To a solution of 2-hydroxyisobutyric acid (0.0567 g, 0.545 mmol) in EtOAC (2.0 mL) at an internal temperature of about 60 °C was added a solution of N,N',N”-{boroxin-2,4,6-trisyltris[[(1R)-3-methylbutane-1,1-diyl]imino(2- oxoethane-2,1-diyl)]}tris(2,5-dichlorobenzamide) (0.200 g, 0.195 mmol) in EtOAC (1.0 mL). The solution was allowed to cool freely until the internal temperature was about 25 °C and the solvent was removed by evaporation to yield 2,5-dichloro-N-(2-{[(1R)-1-(4,4-dimethyl-5-oxo-1,3,2- dioxaborolan-2-yl)-3-methylbutyl]amino}-2-oxoethyl)benzamide (0.255 g, 96%) as a white solid. MS (m / z): [M+Et3N+H] in CH3CN 24 H 39 BC12N3O5 Calc. 530.2; Found 530.0. MS (m / z): [M-H] in CH3CN 18 H 22 BC12N2O5 Calc. 427.1; Found 427.0.

[0864] Example 6: Synthesis of 2,5-dichloro-N-[2-({(1R)-3-methyl-1-[(5R)-4-oxo-5-phenyl-1,3,2- dioxaborolan-2-yl]butyl}amino)-2-oxoethyl]benzamide (I-6)

[0865] To a solution of (R)-mandelic acid (0.168 g, 1.10 mmol) in EtOAC (2.0 mL) at an internal temperature of about 60 °C was added a solution of N,N',N”-{boroxin-2,4,6-trisyltris[[(1R)-3-methylbutane-1,1-diyl]imino(2- oxoethane-2,1-diyl)]}tris(2,5-dichlorobenzamide) (0.382 g, 0.37 mmol) in EtOAC (1.0 mL). The solution was allowed to cool freely until the internal temperature was about 25 °C and the resulting precipitate was collected by filtration to yield 2,5-dichloro-N-[2-({(1R)-3-methyl-1-[(5R)-4-oxo-5-phenyl-1,3,2- dioxaborolan-2-yl]butyl}amino)-2-oxoethyl]benzamide (0.343 g, 65%) as a white solid. 1H NMR (300 MHz, DMSO-d6, δ): 10.88 (s, 1H), 9.22 (m, 1H), 7.68-7.27 (m, 8H), 5.15 (s, 1H), 4.33 (d, J = 6.0 Hz, 2H), 2.8-2.76 (m, 1H), 1.71-1.62 (m, 1H), 1.50-1.28 (m, 2H), 0.89 (m, 6H). MS (m / z) in CH3CN: [M+Et3N+H] C 28 H 39 BC12N3O5Calculated 578.2; found 578.1. MS (m / z) in CH3CN: [M-H] C 22 H 22 BC12N2O5Calculated 475.1; found 475.1.

[0866] Example 7: Synthesis of 2,5-dichloro-N-[2-({(1R)-3-methyl-1-[(4S)-4-methyl-5-oxo-1,3,2- dioxaborolan-2-yl]butyl}amino)-2-oxoethyl]benzamide (I-7)

[0867] To a solution of L-lactic acid (0.675 g, 7.34 mmol) in EtOAC (3.0 mL) at an internal temperature of about 70 °C was added a solution of N,N',N''-{boroxin-2,4,6-trisyl tris[[(1R)-3-methylbutane-1,1- diyl]imino(2-oxoethane-2,1-diyl)]}tris(2,5-dichlorobenzamide) (2.50 g, 2.43 mmol) in EtOAC (7.5 mL). The solution was allowed to cool freely until the internal temperature was about 60 °C. After 30 minutes, heptane (11.5 mL) was added until the solution became cloudy. The suspension was heated until the internal temperature was 70 °C or about 70 °C, at which time a homogenous solution resulted. The solution was cooled at a rate of 0.17 °C / min until the internal temperature was about 30 °C, then allowed to cool freely until the internal temperature was about 0 °C. The resulting precipitate was collected by filtration to yield 2,5-dichloro-N-[2-({(1R)-3-methyl-1-[(4S)-4-methyl-5-oxo-1,3,2-dioxaborolan-2-yl]butyl}amino)-2-oxoethyl]benzamide (2.32 g, 81%) as a white crystalline solid. MS (m / z) in CH3CN: [M+Et3N+H] C 23 H 37 BC12N3O5Calculated 578.2; found 578.1. MS (m / z) in CH3CN: [M-H] C 17 H 20Calculated for BCl2N2O5 413.1 ; found 413.0.

[0868] XRPD data for I-7 is shown in Figure 5

[0869]

[0870]

[0871] Table 4: XRPD data for I-7

[0872] Example 8: Synthesis of 2,5-dichloro-N-[2-({(1R)-3-methyl-1-[(4S)-4-methyl-6- oxo-1,3,2-dioxaborinan-2-yl]butyl}amino)-2-oxoethyl]benzamide (I-8)

[0873] To a solution of (S)-3-hydroxybutanoic acid (0.0598 g, 0.566 mmol) in EtOAC (2.0 mL) at an internal temperature of about 60 °C was added a solution of N,N',N''-{boroxin-2,4,6- triyl tris[[(1R)-3-methylbutane-1,1 -diyl]imino(2-oxoethane-2,1 - diyl)]}tris(2,5-dichlorobenzamide) (0.200 g, 0.195 mmol) in EtOAC (1.0 mL). The solution was allowed to cool freely until the internal temperature was about 25 °C and the solvent was removed by evaporation to yield 2,5-dichloro-N-[2-({(1R)-3-methyl-1-[(4S)-4-methyl-6-oxo-1,3,2-dioxaborinan-2- yl]butyl}amino)-2-oxoethyl]benzamide (0.225 g, 95%) as a white solid. 1 H NMR (300 MHz, DMSO-d6, δ): 10.45 (s, 1H), 9.11 (t, J = 6.0 Hz, 1H), 7.65 (m, 1H), 7.55 (m, 2H), 4.21 (d, J = 6.0 Hz, 2H), 3.98-3.90 (m, 1H), 2.51 (m, 1H), 2.33 (dd, J1= 19.2 Hz, J = 2.7 Hz, 1H), 2.24-2.21 (m, 1H), 1.61-1.52 (m, 1H), 1.33-1.19 (m, 2H), 1.07-1.04 (m, 3H), 0.84 (m, 6H). MS (m / z) in CH3CN: [M+Et3N+H] C 24 Calculated for H39BCl2N3O5 530.2; found 530.0. MS (m / z) in CH3CN: [M-H] C 18 H 22 ​Calculated for BCl2N2O5 441.1 ; found 441.0.

[0874] Example 9: Synthesis of 2,5-dichloro-N-(2-{[(1R)-1-(4,4-dimethyl-6-oxo-1,3,2- dioxaborolan-2-yl)-3-methylbutyl]amino}-2-oxoethyl)benzamide (1-9)

[0875] To a solution of β-hydroxyisovaleric acid (0.0841 g, 0.712 mmol) in EtOAC (2.0 mL) at an internal temperature of about 60 °C was added a solution of N,N',N”-{cycloboronoxane-2,4,6-triyl tris[[(1R)-3-methylbutane-1,1-diyl]imino(2- oxoethane-2,1-diyl)]}tris(2,5-dichlorobenzamide) (0.260 g, 0.253 mmol) in EtOAC (1.0 mL). The solution was allowed to cool freely until the internal temperature was about 26 °C and the solvent was removed by evaporation to yield 2,5-dichloro-N-(2-{[(1R)-1-(4,4-dimethyl-6-oxo-1,3,2-dioxaborolan-2-yl)-3- methylbutyl]amino}-2-oxoethyl)benzamide (0.266 g, 95%) as a white solid. MS (m / z): [M+Et3N+H] C 25 H 41 Calculated for BCl2N3O5 544.3; found 544.0. MS (m / z): [M-H] C 19 H 24 Calculated for BCl2N2O5 441.1 ; found 441.0.

[0876] Example 10: Synthesis of 2,5-dichloro-N-[2-({(1R)-1-[(4S)-4-tert-butyl-5-oxo-1,3,2- dioxaborolan-2-yl]-3-methylbutyl}amino)-2-oxoethyl]-2,5-dichlorobenzamide (1-10)

[0877] To a solution of (S)-2-hydroxy-3,3-dimethylbutanoic acid (0.0712 g, 0.553 mmol) in EtOAC (2.0 mL) at an internal temperature of about 60 °C was added a solution of N,N',N''-{cycloboronoxane-2,4,6-trisyl tris[[(1R)-3-methylbutane-1,1-diyl]imino(2- oxoethane-2,1-diyl)]}tris(2,5-dichlorobenzamide) (0.200 g, 0.195 mmol) in EtOAC (1.0 mL). The solution was allowed to cool freely until the internal temperature was about 25 °C and the solvent was removed by evaporation to yield 2,5-dichloro-N-[2-({(1R)-1-[(4S)-4-tert-butyl-5-oxo-1,3,2- dioxaborinan-2-yl]-3-methylbutyl}amino)-2-oxoethyl]-2,5-dichlorobenzamide (0.245 g, 97%) as a white solid. MS (m / z): [M+Et3N+H] in CH3CN C 26 H 43 Calculated for BCl2N3O5 558.3; found 558.0. MS (m / z): [M-H] in CH3CN C 20 H 26 Calculated for BCl2N2O5 455.1; found 455.0.

[0878] Example 11: Synthesis of 2,5-dichloro-N-[2-({(1R)-1-[(4S)-4-isopropyl-5-oxo-1,3,2- dioxaborinan-2-yl]-3-methylbutyl}amino)-2-oxoethyl]benzamide (1-11)

[0879] To a solution of (S)-2-hydroxy-3-methylbutanoic acid (0.0659 g, 0.558 mmol) in EtOAC (2.0 mL) at an internal temperature of about 60 °C was added a solution of N,N',N''-{cycloboronoxane-2,4,6-trisyl tris[[(1R)-3-methylbutane-1,1-diyl]imino(2- oxoethane-2,1-diyl)]}tris(2,5-dichlorobenzamide) (0.200 g, 0.195 mmol) in EtOAC (1.0 mL). The solution was allowed to cool freely until the internal temperature was about 25 °C and the solvent was removed by evaporation to yield 2,5-dichloro-N-[2-({(1R)-1-[(4S)-4-isopropyl-5-oxo-1,3,2- dioxaborinan-2-yl]-3-methylbutyl}amino)-2-oxoethyl]benzamide (0.246 g, 99%) as a white solid. MS (m / z): [M+Na] in CH3CN C 19 H 25BC12N2Na05found 465.1 ; experimental 465.1. MS (m / z): [M-H] C 19 H 24 BC12N2O5found 441.1 ; experimental 441.0.

[0880] Example 12: Synthesis of 2,5-dichloro-N-[2-({(1R)-1-[(4S)-4-isobutyl-5-oxo-1,3,2- dioxaborolan-2-yl]-3-methylbutyl}amino)-2-oxoethyl]benzamide (1-12)

[0881] To a solution of 2-hydroxyisocaproic acid (0.0752 g, 0.569 mmol) in EtOAC (2.0 mL) at an internal temperature of about 60 °C was added a solution of N,N',N''-{boroxin-2,4,6- triyl tris[[(1R)-3-methylbutane-1,1-diyl]imino(2-oxoethane-2,1-diyl)]} tri(2,5-dichlorobenzamide) (0.200 g, 0.195 mmol) in EtOAC (1.0 mL). The solution was allowed to cool freely until the internal temperature was about 25 °C and the solvent was removed by evaporation to yield 2,5-dichloro-N-[2-({(1R)-1-[(4S)-4-isobutyl-5-oxo-1,3,2-dioxaborolan-2-yl]-3- methylbutyl}amino)-2-oxoethyl]benzamide (0.253 g, 95%) as a white solid. MS (m / z): [M+Na] C 20 H 27 BC12N2Na05found 479.1 ; experimental 479.1. MS (m / z): [M-H] C 20 H 26 BC12N2O5found 455.1 ; experimental 455.1.

[0882] Example 13: Synthesis of 2,5-dichloro-N-(2-{[(1R)-3-methyl-1-(4-oxo-4H-1,3,2- benzenedioxaborolan-2-yl)butyl]amino}-2-oxoethyl)benzamide (1-13)

[0883] To a solution of salicylic acid (0.0758 g, 0.549 mmol) in EtOAC (2.0 mL) at an internal temperature of about 60 °C was added a solution of N,N',N”-{cycloboronoxane-2,4,6-trisyl tris[[(1R)-3-methylbutane-1,1-diyl]imino(2- oxoethane-2,1-diyl)]} tri(2,5-dichlorobenzamide) (0.200 g, 0.195 mmol) in EtOAC (1.0 mL). The solution was allowed to cool freely until the internal temperature was about 25 °C and the resulting precipitate was collected by filtration to yield 2,5-dichloro-N-(2-{[(1R)-3-methyl-1-(4-oxo-4H-1,3,2- benzodioxol-2-yl)butyl]amino}-2-oxoethyl)benzamide (0.198 g, 78%) as a white solid. MS (m / z): [M+Na] C 21 H 21 BC12N2Na05 Found 485.1; 485.1. MS (m / z): [M-H] C 21 H 20 BC12N2O5 Found 461.1; 461.0.

[0884] XRPD data for I-13 is shown in Figure 6 and Table 5.

[0885]

[0886]

[0887] Table 5: XRPD data for I-13

[0888] Example 14: Synthesis of 2,5-dichloro-N-(2-{[(1R)-3-methyl-1-(5-oxo-4,4- diphenyl-1,3,2-dioxaborolan-2-yl)butyl]amino}-2-oxoethyl)benzamide (I-14)

[0889] To a solution of diphenyl glycolic acid (0.126 g, 0.552 mmol) in EtOAC (2.0 mL) at an internal temperature of about 60 °C was added a solution of N,N',N”-{boroxin-2,4,6-trisyl tris[[(1R)-3-methylbutane-1,1-diyl]imino(2- oxoethane-2,1-diyl)]}tris(2,5-dichlorobenzamide) (0.200 g, 0.195 mmol) in EtOAC (1.0 mL). The solution was allowed to cool freely until the internal temperature was about 25 °C and the solvent was removed by evaporation to yield 2,5-dichloro-N-(2-{[(1R)-3-methyl-1-(5-oxo-4,4- diphenyl-1,3,2-dioxaborolan-2-yl)butyl]amino}-2-oxoethyl)benzamide (0.291 g, 95%) as a white solid. MS (m / z): [M+Na] C 28 H 27 Calculated for BCl2N2NaO5 575.1; found 575.2. MS (m / z): [M-H] C 28 H 26 Calculated for BCl2N2O5 551.1; found 551.1.

[0890] Example 15: Synthesis of 2,2'-{2-[(1R)-3-methyl-1-({(2S)-3-phenyl-2- [(pyrazine-2-carbonyl)amino]propanoyl}amino)butyl]-5-oxo-1,3,2-dioxaborinan-4,4- diyl}diacetic acid (I-15)

[0891] To a solution of citric acid (0.257 g, 1.34 mmol) in EtOAC (7.4 mL) at an internal temperature of about 74 °C was added N,N',N”-(boroxin-2,4,6-trisyl tris{[(1R)-3- methylbutane-1,1-diyl]imino[(2S)-1-oxo-3-phenylpropane-1,2-diyl]})tripyrazine-2- carboxamide (0.500 g, 0.455 mmol) as a solid. The resulting solution was allowed to cool freely until the internal temperature was about 25 °C and evaporated to yield 2,2'-{2-[(1R)-3- methyl-1-({(2S)-3-phenyl-2-[(pyrazine-2-carbonyl)amino]propanoyl}amino)butyl]-5-oxo-1,3,2- dioxaborinan-4,4-diyl}diacetic acid (0.730 g, 99%) as a white solid. MS (m / z): [M+Et3N+H] C 31 H 45 Calculated for BN5O9 642.3; found 642.2. MS (m / z): [M-H] C 25H 28 Calculated for BN4O9 539.2; found 539.2.

[0892] Example 16: Synthesis of N-[(1S)-1-benzyl-2-({(1R)-3-methyl-1-[(5R)-4-oxo-5-phenyl- 1,3,2-dioxaborolan-2-yl]butyl}amino)-2-oxoethyl]pyrazine-2-carboxamide (1-16)

[0893] To a solution of (R)-mandelic acid (0.0738 g, 0.485 mmol) in EtOAC (2.0 mL) at an internal temperature of about 60 °C was added N,N',N"-(cycloboroxane-2,4,6- triyl tris{[(1R)-3-methylbutane-1,1-diyl]imino[(2S)-1-oxo-3-phenylpropane-1,2-diyl]}) tripyrazine-2-carboxamide (0.178 g, 0.162 mmol) as a solid. The solution was allowed to cool freely until the internal temperature was about 25 °C and the resulting precipitate was collected by filtration to yield N-[(1S)-1-benzyl-2-({(1R)-3-methyl-1-[(5R)-4-oxo-5-phenyl-1,3,2-dioxaborolan-2-yl]butyl}amino)-2-oxoethyl]pyrazine-2-carboxamide (0.195 g, 80%) as a white solid. MS (m / z): [M+Na] in CH3CN C 27 H 29 Calculated for BN4NaO5 523.2; found 523.2. MS (m / z): [M-H] in CH3CN C 27 H 28 Calculated for BN4O5 499.2; found 499.2.

[0894] Example 17: Synthesis of N-[(1S)-1-benzyl-2-({(1R)-3-methyl-1-[(5R)-4-oxo-5-phenyl- 1,3,2-dioxaborolan-2-yl]butyl}amino)-2-oxoethyl]pyrazine-2-carboxamide (1-17)

[0895] Add solid N,N',N”-(cycloboroxane-2,4,6-triyltri{[(1R)-3-methylbutane-1,1-diyl]imino[(2S)-1-oxo-3-phenylpropane-1,2-diyl]})tripyrazine-2-carboxamide (0.179 g, 0.163 mmol) to a solution of (S)-3-hydroxybutyric acid (0.0509 g, 0.489 mmol) in EtOAC (2.0 mL) at an internal temperature of approximately 60 °C. mmol). The solution was allowed to cool freely until the internal temperature reached approximately 25°C, and the solvent was removed by evaporation to produce N-[(1S)-1-benzyl-2-({(1R)-3-methyl-1-[(4S)-4-methyl-6-oxo-1,3,2-dioxaborane-2-yl]butyl}amino)-2-oxoethyl]pyrazine-2-carboxamide (0.213 g, 96%) as a white solid. MS (m / z) in CH3CN: [M+Na]C 23 H 29 The calculated value of BN4NaO5 is 475.2; the experimental value is also 475.2. MS (m / z) in CH3CN: [MH]C 23 H 28 The calculated value of BN4O5 is 451.2; the experimental value is 451.1.

[0896] Example 18: Preparation of formulations of 4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzoylamino)acetamido)-3-methylbutyl)-6-oxo-1,3,2-dioxaborane-4-carboxylic acid (I-1) for non-enteral or oral administration.

[0897] Formula A: 90 mL of water and 0.08 g of citric acid monohydrate were placed in a container, and 1.5 g of sodium citrate dihydrate was added and stirred until dissolved. 0.142 g of 4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzoylamino)acetamido)-3-methylbutyl)-6-oxo-1,3,2-dioxaborane-4-carboxylic acid (I-1) form 2 was added to this solution, and the mixture was stirred until a solution was obtained. 0.45 g of sodium chloride was added to this solution, and the pH was adjusted to 5.45 using 2N HCl. The final volume of the resulting solution was adjusted to 100 mL with water and filtered through a 0.2 μm PES membrane to produce Formula A, which was stored at -20°C.

[0898] Prepare formulation B as in formulation A, except that the pH is adjusted to pH 6.2 using 2N NaOH.

[0899] Formulation C: A vessel was charged with 90 mL of water and citric acid monohydrate (0.08 g), sodium citrate dihydrate (1.5 g) and propylene glycol (1.0 g) was added and stirred until dissolved. To this solution, 4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetylamino)-3- methylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid (1-1) Form 2 (0.142 g) was added and the mixture was stirred until a solution was obtained. The pH was adjusted to 6.2 using 2N NaOH and the final volume of the resulting solution was adjusted to 100 mL with water and filtered through a 0.2 pm PES membrane to yield Formulation C, which was stored at -20 °C.

[0900] Example 19: Formulation of 4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5- dichlorobenzamido)acetylamino)-3-methylbutyl)-6-oxo-1,3,2-dioxaborinane-4- carboxylic acid (1-1) prepared extemporaneously for parenteral or oral administration

[0901] Stock Formulation Vehicle: A vessel was charged with about 160 mL of water and citric acid monohydrate (0.714 g) and sodium citrate dihydrate (2.24 g) was added and stirred until dissolved. To this solution, propylene glycol (2.0 g) was added and the mixture was stirred until a homogeneous solution was obtained. The final pH was pH 5.14. The final weight of the resulting solution was adjusted to 200 g with water (assuming a density of 1 g / mL) and filtered through a 0.2 pm PES membrane filter unit and stored at a temperature between about 2 °C and about 8 °C.

[0902] Stock Formulation (1 mg / mL): To a vessel was added 0.105 grams (about 95.4% purity) of N,N',N''-{cycloboronane-2,4,6-trisyltris[[(1R)-3-methylbutane-1,1-diyl]imino(2- oxoethane-2,1-diyl)]}tris(2,5-dichlorobenzamide). To this was added about 90 g of Stock Formulation Vehicle and the resulting mixture was stirred protected from light for 48 hours. The final pH was pH 5.12. The final weight of the resulting solution was adjusted to 100 g with Stock Formulation Vehicle (assuming a density of 1 g / mL) and filtered through a 0.2 pm PES membrane filter unit and stored at a temperature between about 2 °C and about 8 °C protected from light.

[0903] Formulation D: The Stock Formulation was diluted with Stock Formulation Vehicle to concentrations of 0.05 mg / mL and 0.1 mg / mL prior to use.

[0904] Formulation E: The Stock Formulation was diluted with 0.9% sodium chloride solution to concentrations of 0.05 mg / mL and 0.1 mg / mL prior to use.

[0905] Example 20: 20S Proteasome Assay

[0906] In 384-well black microtiter plates, 25 μL of assay buffer containing human PA28 activator (Boston Biochem, final 12 nM) with Ac-WLA-AMC (β5 selective substrate) (final 15 μM) was added to 1 μL of test compound dissolved in DMSO at 37°C, followed by 25 μL of assay buffer containing human 20S proteasome (Boston Biochem, final 0.25 nM) at 37°C. Assay buffer consisted of 20 mM HEPES, 0.5 mM EDTA, and 0.01% BSA (pH 7.4). Reactions were followed by a BMG Galaxy microplate reader (37°C, excitation 380 nm, emission 460 nm, gain 20). Percent inhibition was calculated relative to 0% inhibition (dMSO) and 100% inhibition (10 μM bortezomib) controls.

[0907] Example 21: Anti-proliferative Assay

[0908] HCT-116 (1000) or other tumor cells in 100 μL of appropriate cell culture medium (McCoy's 5A for HCT-116, Invitrogen) supplemented with 10% fetal bovine serum (Invitrogen) were seeded in wells of a 96-well cell culture plate and incubated at 37°C overnight. Test compounds were added to the wells and the plate was incubated at 37°C for 96 hours. MTT or WST reagent (10 μL, Roche) was added to each well as described by the manufacturer and incubated at 37°C for 4 hours. For MTT, the metabolized dye was solubilized overnight according to the manufacturer's instructions (Roche). The optical density of each well was read using a spectrophotometer (Molecular Devices) at 595 nm (primary) and 690 nm (reference) for MTT and at 450 nm for WST. For MTT, the reference optical density value was subtracted from the primary wavelength value. Percent inhibition was calculated using the DMSO control set to 100% of the value.

[0909] Example 22: In vivo Tumor Efficacy Model

[0910] Freshly isolated HCT-116 (2-5 x 10 6) or other tumor cells are injected subcutaneously into the right flank of female CD-1 nude mice (5-8 weeks old, Charles River) under aseptic conditions. Alternatively, certain xenograft models require serial passage of tumor fragments. In these cases, small fragments of tumor tissue (approximately 1 mm 3 ) are implanted subcutaneously into the right flank of anesthetized (3-5% isoflurane / oxygen mixture) C.B-17 / SCID mice (5-8 weeks old, Charles River). Beginning on day 7 post-inoculation, tumors are measured twice weekly using vernier calipers. Tumor volume is calculated using standard procedures (0.5 x (length x width 2 )2). When tumors reach a volume of approximately 200 mm 3 , mice are randomized into treatment groups and drug therapy is initiated. Dosing and schedule for each experiment are determined based on previous results from pharmacokinetic / pharmacodynamic and maximum tolerated dose studies. Control groups receive vehicle without any drug. Test compounds (100-200 μL) are typically administered via intravenous (27 gauge needle), oral (20 gauge gavage needle), or subcutaneous (27 gauge needle) at various doses and schedules. Tumor size and body weight are measured twice weekly and studies are terminated when control tumors reach a volume of approximately 2000 mm 3 .

[0911] Example 23: Synthesis of N-((S)-1-((R)-3-methyl-1-(4-oxo-4H-benzo[d][1,3,2]dioxaborolan-2-yl)butylamino)-1-oxo-3-phenylpropan-2-yl)pyrazine-2-carboxamide (1-19)

[0912] A mixture of N,N',N"-(cycloboroxane-2,4,6-trisyltris{[(1R)-3-methylbutane-1,1- diyl]imino[(2S)-1-oxo-3-phenylpropane-1,2-diyl]})tripyrazine-2-carboxamide (0.250 g, 0.228 mmol) and salicylic acid (269.6 mg, 0.68 mmol) was mixed in EtOAC (10 mL). The mixture was heated to form a solution. The solution was allowed to cool freely until the internal temperature was about 25 °C. Heptane (16 mL) was added. A white solid precipitated and the resulting slurry was stirred at ambient temperature for 3 hours. The slurry was filtered to collect the solid N-((S)-1-((R)-3-methyl-1-(4-oxo-4H-benzo[d][1,3,2]dioxaborolan-2-yl)butylamino)-1-oxo-3-phenylpropan-2-yl)pyrazine-2-carboxamide (0.249 g, 75%). MS (m / z): [M+H] C 26 H 28Calculated for BN4O5 487.2153; found 487.3.

[0913] Example 24: Synthesis of 2-((S)-2-((R)-3-methyl-l-((S)-3-phenyl-2- (pyrazine-2-carboxamido)propanamido)butyl)-5-oxo-l,3,2-dioxaborolan-4- yl)acetic acid (1-20)

[0914] A mixture of N,N',N"-(cycloboroxane-2,4,6-trisyl tris{[(1R)-3-methylbutane-1,1- diyl]imino[(2S)-1-oxo-3-phenylpropane-1,2-diyl]})tripyrazine-2-carboxamide (0.500 g, 0.455 mmol) and L-malic acid (213.6 mg, 0.55 mmol) was mixed in THF (5 mL). The mixture was heated to form a solution. The solution was allowed to cool freely until the internal temperature was about 25 °C. A white solid precipitated out and the resulting slurry was stirred at ambient temperature for 1 hour. The slurry was filtered to collect the solid 2-((S)-2-((R)-3-methyl-l-((S)-3-phenyl-2- (pyrazine-2-carboxamido)propanamido)butyl)-5-oxo-l,3,2-dioxaborolan-4- yl)acetic acid (0.625 g, 95%). MS (m / z): [M+H] C 23 H 28 Calculated for BN4O7 483.2051; found 483.2.

[0915] Example 25: Synthesis of 2-((R)-2-((R)-3-methyl-l-((S)-3-phenyl-2- (pyrazine-2-carboxamido)propanamido)butyl)-5-oxo-l,3,2-dioxaborolan-4- yl)acetic acid (1-21)

[0916] A mixture of N,N',N"-(cycloboroxane-2,4,6-trisyl tris{[(1R)-3-methylbutane-1,1- diyl]imino[(2S)-1-oxo-3-phenylpropane-1,2-diyl]})tripyrazine-2-carboxamide (0.305 g, 0.278 mmol) and D-malic acid (130.3 mg, 0.33 mmol) was mixed in acetone (3 mL). The mixture was heated to form a solution. The solution was allowed to cool freely until the internal temperature was about 25 °C. A white solid precipitated out and the resulting slurry was stirred at ambient temperature for 3 hours. The slurry was filtered to collect the solid 2-((R)-2-((R)-3-methyl-l-((S)-3-phenyl-2- (pyrazine-2-carboxamido)propanamido)butyl)-5-oxo-l,3,2-dioxaborolan-4- yl)acetic acid (0.410 g, 100%). [M+H] C 23 H28 Calculated for BN4O7 483.2051 ; found 483.2.

[0917] Example 26: Synthesis of (R)-2-hydroxy-2-((R)-2-((R)-3-methyl-l-((S)-3- phenyl-2-(pyrazine-2-carboxamido)propanamido)butyl)-5-oxo-l,3,2-dioxaborolan-4- yl)acetic acid (1-22)

[0918] A mixture of N,N',N"-(cycloboroxane-2,4,6-trisyl tris{[(lR)-3-methylbutane-l,l- diyl]imino[(2S)-l-oxo-3-phenylpropane-l,2-diyl]})tripyrazine-2-carboxamide (0.270 g, 0.246 mmol) and L-tartaric acid (149.5 mg, 0.33 mmol) was mixed in acetone (3 mL). The mixture was heated to form a solution. The solution was allowed to cool freely until the internal temperature was about 25 °C. Heptane (2.5 mL) was added. A white solid precipitated and the resulting slurry was stirred at ambient temperature for 1.5 hours. The slurry was filtered to collect the solid (R)-2-hydroxy-2-((R)-2-((R)-3-methyl-l-((S)-3-phenyl-2- (pyrazine-2-carboxamido)propanamido)butyl)-5-oxo-l,3,2-dioxaborolan-4-yl)acetic acid (0.388 g) which also contained dimeric material. MS (m / z): [M+H] C 23 H 28 Calculated for BN4O8 499.2000; found 499.2.

[0919] Example 27: Synthesis of (S)-2-hydroxy-2-((S)-2-((R)-3-methyl-l-((S)-3- phenyl-2-(pyrazine-2-carboxamido)propanamido)butyl)-5-oxo-l,3,2-dioxaborolan-4- yl)acetic acid (1-23)

[0920] A mixture of N,N',N"-(cycloboroxane-2,4,6-triyl tri{[(1R)-3-methylbutane-1,1- diyl]imino[(2S)-1-oxo-3-phenylpropane-1,2-diyl]})tripyrazine-2-carboxamide (0.180 g, 0.164 mmol) and D-tartaric acid (147.5 mg, 0.33 mmol) was mixed in acetone (4 mL). The mixture was heated to form a solution. The solution was allowed to cool freely until the internal temperature was about 25 °C. Heptane (8 mL) was added. The mixture was evaporated to yield (S)-2-hydroxy-2-((S)-2-((R)-3-methyl-1-((S)-3-phenyl-2- (pyrazine-2-carboxamido)propanamido)butyl)-5-oxo-1,3,2-dioxaborolan-4-yl)acetic acid (0.447 g) which also contained dimeric material. MS (m / z): [M+H] C 23 H 28 Calculated for BN4O8 499.2000; found 499.2.

[0921] Example 28: Pharmaceutical Composition 1

[0922] Capsule compositions are shown in Table 7 below.

[0923] Component Function mg / capsule Form 2 of compound of formula (I-1) 0.29 Microcrystalline cellulose (low moisture) Filler 89.71 Total capsule content weight, mg 90.00 White opaque gelatin capsules, size 4

[0924] Table 7: Capsule compositions

[0925] Example 29: Pharmaceutical Composition 2

[0926] Capsule compositions are shown in Table 8 below.

[0927] Component Function mg / capsule Form 2 of compound of formula (I-1) 0.29 Silicified microcrystalline cellulose Filler 109.71 Total capsule content weight, mg 110.00 White opaque gelatin capsules, size 4

[0928] Table 8: Capsule compositions

[0929] Example 30: Pharmaceutical Composition 3

[0930] Capsule compositions are shown in Table 9 below.

[0931] Component Function mg / capsule Form 2 of compound of formula (I-1) 0.29 Microcrystalline cellulose (low moisture) Filler 88.81 Magnesium stearate Lubricant 0.90 Total capsule content weight, mg 90.00 White opaque gelatin capsules, size 4

[0932] Table 9: Capsule compositions

[0933] Example 31: Pharmaceutical Composition 4

[0934] Capsule compositions are shown in Table 10 below.

[0935] Component Function mg / capsule Form 2 of compound of formula (I-1) 0.29 Microcrystalline cellulose Filler 78.91 Magnesium stearate Lubricant 0.80 Total capsule content weight, mg 80.00 White opaque gelatin capsules, size 4

[0936] Table 10: Capsule compositions

[0937] Example 32: Pharmaceutical Composition 5

[0938] Capsule composition is shown in Table 11 below.

[0939] Component Function mg / capsule Form 2 of compound of formula (I-1) 0.29 Microcrystalline cellulose (low moisture) Filler 84.71 Total capsule content weight, mg 85.00 White opaque gelatin capsules, size 4

[0940] Table 11: Capsule composition

[0941] Example 33: Pharmaceutical composition 6

[0942] Capsule composition is shown in Table 12 below.

[0943] Component Function mg / capsule Form 2 of compound of formula (I-1) 0.72 Microcrystalline cellulose (low moisture) Filler 119.28 Total capsule content weight, mg 120.00 White opaque gelatin capsules, size 4

[0944] Table 12: Capsule composition

[0945] Example 34: Pharmaceutical composition 7

[0946] Capsule composition is shown in Table 13 below.

[0947] Component Function mg / capsule Form 2 of compound of formula (I-1) 2.89 Microcrystalline cellulose (low moisture) Filler 147.11 Total capsule content weight, mg 150.00 Dark green gelatin capsules, size 3

[0948] Table 13: Capsule composition

[0949] Example 35: Pharmaceutical composition 8

[0950] Composition is shown in Table 14 below.

[0951]

[0952]

[0953] Table 14: Batch composition

[0954] The batch was prepared according to the following method:

[0955] 1) Sieve microcrystalline cellulose (NF) (Item 1) through a 40-micron sieve. XLM90; low moisture) (Item 2).

[0956] 2) Add the sieved material from Step 1 to a PK blender and blend for 2 minutes.

[0957] 3) Weigh the Form 2 of the compound of Formula (I-1) (Item 1) that has been sieved through a 60-micron sieve.

[0958] 4) Combine Form 2 of the compound of Formula (I-1) from Step 3 and microcrystalline cellulose (NF) (Item 3) in a polyethylene bag and shake the polyethylene bag; then pass the contents of the polyethylene bag through the same 40-micron sieve as used in Step 1.

[0959] ​5) Add the material from step 4) to the PK blender and blend for 15 minutes.

[0960] 6) Sieve the microcrystalline cellulose (NF) (Item 3) through the same 40 micron screen, XLM90; low moisture) (Item 4) into the same polyethylene bag used in steps 4) and 6) and shake in the polyethylene bag.

[0961] 7) Add the material from step 6) to the PK blender which still contains the material from step 5) and blend for 10 minutes.

[0962] 8) Sieve the microcrystalline cellulose (NF) (Item 3) through the same 40 micron screen, XLM90; low moisture) (Item 5) into the same polyethylene bag used in steps 4), 6) and 8) and shake in the polyethylene bag.

[0963] 9) Add the material from step 8) to the PK blender which still contains the material from steps 5) and 7) and blend for 10 minutes.

[0964] 10) Sieve the microcrystalline cellulose (NF) (Item 3) through the same 40 micron screen, XLM90; low moisture) (Item 6) into the same polyethylene bag used in steps 4), 6) and 8) and shake in the polyethylene bag.

[0965] 11) Add the material from step 10) to the PK blender which still contains the material from steps 5), 7) and 9) and blend for 10 minutes.

[0966] 12) Encapsulate the material from the blender in size 4 white opaque gelatin capsules using the In-Cap system.

[0967] 13) Dust the capsules and weight pick.

[0968] Example 36: Pharmaceutical Composition 9

[0969] The capsule composition is shown in Table 15 below.

[0970] Component Function mg / capsule Form 2 of compound of formula (I-1) 0.3 Microcrystalline cellulose (low moisture) Filler 122.825 Total capsule content weight, mg Orange gelatin capsules, size 2, Swedish type 1.25 Component Function 0.625 mg / capsule 125.00 Form 2 of compound of formula (I-1)

[0971] Table 15: Capsule Composition

[0972] Example 37: Pharmaceutical Composition 10

[0973] The capsule composition is shown in Table 16 below.

[0974] Pre-gelatinized starch (starch 1500) Filler Talc Glidant 0.3 Magnesium stearate Lubricant 124.7 Total capsule content weight, mg 125.00 White opaque gelatin capsules, size 4

[0975] Table 16: Capsule Composition

[0976] Example 38: Pharmaceutical composition 11

[0977] Capsule compositions are shown in Table 17 below.

[0978]

[0979] Table 17: Capsule compositions

[0980] Example 39: Pharmaceutical composition 12

[0981] Capsule compositions are shown in Table 18 below.

[0982]

[0983] Table 18: Capsule compositions

[0984] Example 40: Pharmaceutical composition 13

[0985] Capsule compositions are shown in Table 19 below.

[0986]

[0987] Table 19: Capsule compositions

[0988] Example 41: Pharmaceutical composition 14

[0989] Capsule compositions are shown in Table 20 below.

[0990]

[0991] Table 20: Capsule compositions

[0992] Example 42: Pharmaceutical composition 15

[0993] Capsule compositions are shown in Table 21 below.

[0994]

[0995] Table 21: Capsule compositions

[0996] Example 43: Pharmaceutical composition 16

[0997] Batch compositions are shown in Table 22 below.

[0998]

[0999] Table 22: Batch compositions

[1000] The batches were prepared according to the following method:

[1001] 1) Sieve microcrystalline cellulose (NF) (Item 1) through a 40-micron screen. XLM 90; low moisture) (Item 2).

[1002] 2) Add the sieved material from step 1 to the PK mixer and mix for 2 minutes.

[1003] 3) Weigh the compound form 2 (item 1) of formula (I-1) that has been sieved through a 60-micron sieve.

[1004] 4) Compound form 2 of formula (I-1) from step 3 and microcrystalline cellulose (NF) XLM90 (low moisture) (item 3) is incorporated into a polyethylene bag and the polyethylene bag is shaken; then the contents of the polyethylene bag are passed through the same 40-micron sieve as used in step 1.

[1005] 5) Add the material from step 4) to the PK mixer and mix for 15 minutes.

[1006] 6) Microcrystalline cellulose (NF) is sieved through the same 40-micron sieve. XLM90 (low moisture) (item 4), transfer to the same polyethylene bag used in step 4) and shake in the polyethylene bag.

[1007] 7) Talc (item 7) and sodium citrate (item 8) were separated by sieving through a 40-micron sieve.

[1008] 8) Add the substances from steps 6) and 7) to the PK mixer that still contains the substances from step 5) and mix for 10 minutes.

[1009] 9) Microcrystalline cellulose (NF) is sieved through the same 40-micron sieve. XLM90 (low moisture) (item 5), transfer to the same polyethylene bag used in steps 4) and 6) and shake in the polyethylene bag.

[1010] 10) Add the substance from step 9) to the PK mixer that still contains the substances from steps 5) and 8) and mix for 10 minutes.

[1011] 11) Microcrystalline cellulose (NF) is sieved through the same 40-micron sieve. XLM90 (low moisture) (item 6), transfer to the same polyethylene bag used in steps 4), 6) and 9) and shake in the polyethylene bag.

[1012] 12) Add the substance from step 11) to the PK mixer that still contains the substances from steps 5), 8) and 10) and mix for 10 minutes.

[1013] 13) Magnesium stearate was sieved through the same 40-micron sieve (item 9).

[1014] 14) Add the substance from step 13) to the PK mixer that still contains the substances from steps 5), 8), 10) and 12) and mix for 5 minutes.

[1015] 15) Use the Profill system to encapsulate the material from the blender into No. 4 white opaque gelatin capsules.

[1016] 16) Remove dust from the capsules and sort them by weight.

[1017] Example 44: Pharmaceutical Composition 17

[1018] The composition of the batch is shown in Table 23 below.

[1019]

[1020] Table 23: Composition of Batch Materials

[1021] The batch material is prepared according to the following method:

[1022] 1) Microcrystalline cellulose (NF) is sieved through a 40-micron sieve. XLM90; Low Moisture) (Item 2).

[1023] 2) Add the sieved material from step 1 to the small PK mixer and mix for 2 minutes.

[1024] 3) Weigh the compound form 2 (item 1) of formula (I-1) that has been sieved through a 60-micron sieve.

[1025] 4) Compound form 2 of formula (I-1) from step 3 is combined with microcrystalline cellulose (NF) ( XLM90 (low moisture) (item 3) is combined and then passed through the same 40-micron sieve as used in step 1).

[1026] 5) Add the material from step 4) to the small PK mixer and mix for 30 minutes.

[1027] 6) Microcrystalline cellulose (NF) is sieved through the same 40-micron sieve. XLM90 (low moisture) (item 4) and talc (item 5).

[1028] 7) Add the substance from step 6) to the small PK mixer that still contains the substance from step 5) and mix for 15 minutes.

[1029] 8) Microcrystalline cellulose (NF) is sieved through the same 40-micron sieve. XLM90 (low moisture) (item 6), transfer to a second, larger PK mixer and mix for 2 minutes.

[1030] 9) The contents from the small PK blender in steps 5) and 7) are injected into a polyethylene bag and then transferred to the larger PK blender in step 8).

[1031] 10) Talc (item 7) and microcrystalline cellulose (NF) are separated by sieving through the same 40-micron sieve. XLM90; Low Moisture) (Item 8).

[1032] 11) Add half of the material from step 10) to the small PK mixer from steps 5) and 7), mix for 3 minutes, transfer to the same polyethylene bag used in step 9), and shake in the polyethylene bag.

[1033] 12) Add the material from step 11) to a larger PK mixer that still contains the material from steps 8) and 9).

[1034] 13) Add the other half of the material from step 10) to the small PK mixer from steps 5), 7) and 11), mix for 3 minutes, transfer to the same polyethylene bag used in steps 9) and 11), and shake in the polyethylene bag.

[1035] 14) Add the material from step 13) to a larger PK mixer that still contains the materials from steps 8), 9) and 12) and mix for 10 minutes.

[1036] 15) Microcrystalline cellulose (NF) is sieved through the same 40-micron sieve. XLM90 (low moisture) (item 9), transfer to the same polyethylene bag used in steps 9), 11) and 13) and shake in the polyethylene bag.

[1037] 16) Add the material from step 15) to the same larger PK mixer that still contains the materials from steps 8), 9), 12) and 14) and mix for 10 minutes.

[1038] 17) Microcrystalline cellulose (NF) is sieved through the same 40-micron sieve. XLM90; Low Moisture) (Item 10).

[1039] 18) Add the material from step 17) to the same larger PK mixer that still contains the materials from steps 8), 9), 12), 14) and 16) and mix for 10 minutes.

[1040] 19) Microcrystalline cellulose (NF) is sieved through the same 40-micron sieve. XLM90; Low Moisture) (Item 11).

[1041] 20) Add the material from step 19) to the same larger PK mixer that still contains the material from steps 8), 9), 12), 14), 16) and 18) and mix for 10 minutes.

[1042] 21) Magnesium stearate was sieved through the same 40-micron sieve (item 12).

[1043] 22) Add the material from step 21) to the same larger PK mixer that still contains the material from steps 8), 9), 12), 14), 16), 18) and 20) and mix for 5 minutes.

[1044] 23) The material from the blender was encapsulated in a white, opaque gelatin capsule (No. 4) using the Incap system.

[1045] 24) Remove dust from the capsules and sort them by weight.

[1046] Example 45: Pharmaceutical Composition 18

[1047] The composition of the batch is shown in Table 24 below.

[1048]

[1049] Table 24: Composition of Batch Materials

[1050] The batch material is prepared according to the following method:

[1051] 1) Microcrystalline cellulose (NF) is sieved through a 40-micron sieve. XLM90 (low moisture) (item 2) and add to the high shear mixer.

[1052] 2) Sieve compound form 2 (I-1) through a 60-micron sieve and weigh it (item 1) and add it to the same high-shear mixer from step 1).

[1053] 3) Microcrystalline cellulose (NF) is sieved through the same 40-micron sieve. XLM90 (low moisture) (item 3) and added to the same high-shear mixer from steps 1) and 2).

[1054] 4) Run the high-shear mixer from steps 1), 2), and 3) for 4 minutes.

[1055] 5) Use the InCap system to encapsulate the material from the high-shear mixer into No. 4 white opaque gelatin capsules.

[1056] 6) Remove dust from the capsules and sort them by weight.

[1057] Example 46: Lyophilized Powder 1

[1058] In a clean container, prepare a 40% t-butanol / 60% water for injection solution by warming the required amount of t-butanol to 35°C and adding water for injection. Cool the solution to 15-30°C. Add a portion of the required amount of the t-butanol / water solution (60% of the total batch size) to a premix container. Reserve about 40% of the solution for rinsing. With agitation, add citric acid (30% of the batch size) to the premix container. Rinse the container with the reserved t-butanol / water solution and add the rinse to the premix container. Stir the mixture until the citric acid is completely dissolved. With agitation, add sodium citrate (30% of the batch size) to the premix container. Rinse the container with the reserved t-butanol / water solution and add the rinse to the premix container. Stir the mixture until the sodium citrate is completely dissolved. With agitation, add N-(2-pyrazinyl)carbonyl-L-phenyl-L-leucine boronic acid (VIII-15) to the premix container. Rinse the container with the reserved t-butanol / water solution and add the rinse to the premix container. Stir the mixture until the boronic acid is completely dissolved. Transfer the citric acid, sodium citrate and boronic acid mixture from the premix container to the main mix container. Rinse the premix container with water for injection and add the rinse to the main mix container. With agitation, add citric acid (70% of the batch size) to the main container. Rinse the container with water and add the rinse to the main container. Stir the mixture until the citric acid is completely dissolved. With agitation, add sodium citrate (70% of the batch size) to the main container. Rinse the container with water and add the rinse to the premix container. Stir the mixture until the sodium citrate is completely dissolved. Add glycine to the main container and rinse the residual glycine with water and add the rinse to the main container. Stir the mixture until the glycine is completely dissolved. Add sufficient water to reduce the total alcohol content to 4.7% v / v. Filter the mixture through a 0.22 μιη filter. Place an aliquot of the filtered solution in a vial. Seal the vial with a lyophilization vial stopper and place on a lyophilization chamber shelf maintained at 20°C. Cool the lyophilization chamber shelf to -45°C using an appropriate temperature ramp rate and maintain at this temperature for 200 minutes. Warm the shelf to -20°C using an appropriate temperature ramp rate and maintain at this temperature for 480 minutes. Cool the shelf again to -45°C using an appropriate temperature ramp rate and maintain at this temperature. After 200 minutes, evacuate the lyophilization chamber and adjust the chamber pressure to 150 microns with nitrogen gas. Warm the shelf to -25°C using an appropriate temperature ramp rate and maintain at this temperature for 3000 minutes. After each product thermocouple reading is -25°C or above, warm the shelf to 27°C and maintain at this temperature for 600 minutes. At the end of the final drying stage, restore the chamber pressure using nitrogen gas and remove the vial. The pre-lyophilized solution contains: 52 mM citrate, 3% glycine, 4.7% t-butanol (as shown in Table 25 below).

[1059] Component Function mg / capsule Form 2 of compound of formula (I-1) Pre-gelatinized starch (starch 1500) Filler 1. Total capsule content weight, mg 0.001g 2.6 0.300g White opaque gelatin capsules, size 4 2. Number 0.00382g 18.2 1.147g Component 3. Amount / mL 0.00994g 33.8 2.982g mM 4. Batch 0.03g 399.6 9.0g Amount per vial 5. Compound (VIII-15) 3.5 mg Citric acid monohydrate, USP / EP 13.37 mg Sodium citrate dihydrate, USP / EP 6. 34.79 mg Glycine, USP / EP 105 mg Tert-butanol, ACS grade Not applicable 7. Not applicable 14.1 mL 0.1645 mL Water for injection, USP / EP Not applicable 8. Not applicable Fill to batch volume Not applicable 5.08 Total volume

[1060] Table 25: Batch Composition

[1061] Example 47: Lyophilized Powder 2

[1062] Prepared as described in Example 46. The pre-lyophilization solution contained: 52 mM citrate; 3% glycine; and 4.7% t-butanol (as shown in Table 26 below).

[1063] Not applicable Not applicable 300 mL 3.5 mL Final measured pH Not applicable 1. Not applicable 0.001g 2.6 0.300g Not applicable 2. Number 0.00168g 8.0 0.504g Component 3. Amount / mL 0.0129g 44.0 3.882g mM 4. Batch 0.03g 399.6 9.0g Amount per vial 5. Compound (VIII-15) 3.5 mg Citric acid monohydrate, USP / EP 5.88 mg Sodium citrate dihydrate, USP / EP 6. 45.15 mg Glycine, USP / EP 105 mg Tert-butanol, ACS grade Not applicable 7. Not applicable 14.1 mL 0.1645 mL Water for injection, USP / EP Not applicable 8. Not applicable Fill to batch volume Not applicable 5.84 Total volume

[1064] Table 26: Batch Composition

[1065] Example 48: Lyophilized Powder 3

[1066] The formulation was prepared as described in Example 46 except the lyophilization cycle was changed. The vials were sealed with lyophilization stoppers and placed on a lyophilization chamber shelf maintained at 20°C. The lyophilization chamber shelf was cooled to -45°C using an appropriate temperature ramp rate and held at this temperature for 200 minutes. The shelf was warmed to -20°C using an appropriate temperature ramp rate and maintained at this temperature for 480 minutes. The shelf was again cooled to -45°C using an appropriate temperature ramp rate and held at this temperature. After 200 minutes, the lyophilization chamber was evacuated and the chamber pressure was adjusted to 150 microns with nitrogen gas. The chamber shelf was warmed to -15°C using an appropriate temperature ramp rate and held at this temperature for 2700 minutes. After each product thermocouple reading was -15°C or above, the shelf was warmed to 37°C and maintained at this temperature for 300 minutes. At the end of the final drying stage, the chamber pressure was restored using nitrogen gas and the vials were sealed and removed. The pre-lyophilization solution contained: 52 mM citrate; 3% glycine; and 4.7% t-butanol (as shown in Table 27 below).

[1067] Not applicable Not applicable 300 mL 3.5 mL Final measured pH Not applicable Not applicable Not applicable Number Component Amount / mL mM Batch Amount per vial mM Batch Amount per vial 1. Compound (VIII-15) 0.001g 2.6 1.0g 3.5 mg 2. Citric acid monohydrate, USP / EP 0.00382g 18.2 3.82g 13.37 mg 3. Sodium citrate dihydrate, USP / EP 0.00994g 33.8 9.94g 34.79 mg 4. Glycine, USP / EP 0.03g 399.6 30.0g 105 mg 5. Tert-butanol, ACS grade Not applicable Not applicable 47.0 mL 0.1645 mL 6. Water for injection, USP / EP Not applicable Not applicable Fill to batch volume Not applicable 7. Total volume Not applicable Not applicable 1000 mL 3.5 mL 8. Final measured pH Not applicable Not applicable 5.05 Not applicable

[1068] Table 27: Batch Composition

[1069] Example 49: Lyophilized Powder 4

[1070] Water for injection is charged to a clean container. Citric acid and sodium citrate are added and stirred until dissolved. To this solution, N-(2-pyrazinyl)carbonyl-L-phenyl-L-leucine borate (VIII-15) is added and stirred until dissolved. Glycine is added to the container and the residual glycine is rinsed with water and the rinse is added to the main container. The mixture is stirred until the glycine is completely dissolved. Enough water is added to bring the batch volume. The mixture is filtered through a 0.22 μιη filter. An aliquot of the filtered solution is placed in a vial. The vial is sealed with a lyophilization vial stopper and placed on a lyophilization chamber shelf maintained at 20 °C. The lyophilization chamber shelf is cooled to -45 °C using a suitable temperature ramp rate and maintained at this temperature for 200 minutes. The shelf is warmed to -20 °C using a suitable temperature ramp rate and maintained at this temperature for 480 minutes. The shelf is again cooled to -45 °C using a suitable temperature ramp rate and maintained at this temperature. After 200 minutes, the lyophilization chamber is evacuated and the chamber pressure is adjusted to 150 microns with nitrogen gas. The chamber shelf is warmed to -25 °C using a suitable temperature ramp rate and maintained at this temperature for 3000 minutes. After each product thermocouple reading is -25 °C or above, the shelf is warmed to 27 °C and maintained at this temperature for 600 minutes. At the end of the final drying stage, the chamber pressure is restored using nitrogen gas and the vial is sealed and removed. The pre-lyophilized solution contains: 52 mM citrate and 3% glycine (as shown in Table 28 below).

[1071] Component Amount / milliliter mM Batch Amount per vial 1. Compound (VIII-15) 0.001g 2.6 0.30g 3.5 mg 2. Citric acid monohydrate, USP / EP 0.004097g 19.5 1.229g 14.34 mg 3. Sodium citrate dihydrate, USP / EP 0.009557g 32.5 2.867g 33.45 mg 4. Glycine, USP / EP 0.03g 399.6 9.0g 105 mg 5. Water for injection, USP / EP Not applicable Not applicable Fill to batch volume Not applicable 6. Total volume Not applicable Not applicable 300 mL 3.5 mL 7. Final measured pH Not applicable Not applicable 4.90 Not applicable

[1072] Table 28: Batch Composition

[1073] Example 50: Lyophilized Powder 5

[1074] Prepared as described in Example 49. The pre-lyophilized solution contains: 52 mM citrate and 3% glycine (as shown in Table 29 below). In this example, the pH of the pre-lyophilized solution is adjusted to a final measured pH value by the addition of 2N HC1.

[1075]

[1076] Table 29: Batch Composition

[1077] Example 50: Lyophilized Powder 6

[1078] Water for injection is charged to a clean container. Citric acid and sodium citrate are added and stirred until dissolved. To this solution, 4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5- dichlorobenzamido)acetylamino)-3-methylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid (1-1) is added and stirred until dissolved. Glycine is added to the container and the residual glycine is rinsed in with water and the rinse added to the main container. The mixture is stirred until the glycine is completely dissolved. Enough water is added to bring the batch volume. The mixture is filtered through a 0.22 μιη filter. An aliquot of the filtered solution is placed in a sterile vial. The vial is sealed with a lyophilization stopper and placed on a lyophilization chamber shelf maintained at 20°C. The lyophilization chamber shelf is cooled to -45°C using an appropriate temperature ramp rate and then held at this temperature for 200 minutes. The shelf is warmed to -20°C using an appropriate temperature ramp rate and then maintained at this temperature for 480 minutes. The shelf is again cooled to -45°C using an appropriate temperature ramp rate and maintained at this temperature. After 200 minutes, the lyophilization chamber is evacuated and the chamber pressure is adjusted to 150 microns with nitrogen gas. The chamber shelf is warmed to -25°C using an appropriate temperature ramp rate and held at this temperature for 3000 minutes. After each product thermocouple reading is -25°C or above, the shelf is warmed to 27°C and maintained at this temperature for 600 minutes.

[1079] At the end of the final drying phase, the chamber pressure is restored using nitrogen gas, and the vial is sealed and removed. The composition of the pre-lyophilized solution is 55 mM citrate; and 3% glycine (as shown in Table 30 below).

[1080]

[1081] Table 30: Batch Composition

[1082] Example 51: Reconstitution of Lyophilized Powder

[1083] The cake structure, cake stability, residual solvent, and residual moisture of a lyophilized powder (e.g., as prepared in Examples 46-50) is analyzed using XRPD, DSC, gas chromatography, and Karl Fisher, respectively. The lyophilized powder is reconstituted with an appropriate amount of sterile water for injection or sterile 0.9% sodium chloride solution for injection. The reconstituted solution is analyzed for purity and ester percentage using HPLC and NMR.

[1084] Example 52: Formulation of 4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5- dichlorobenzamido)acetylamino)-3-methylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid (1-1) Form 2 for Parenteral or Oral Administration

[1085] Water was charged to a vessel and citric acid monohydrate and sodium citrate dihydrate were added and stirred until dissolved. To this solution was added 4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetylamino)-3-methylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid (1-1) Form 2 and the mixture was stirred until a solution was obtained. To this solution was added sodium chloride and stirred until dissolved. Enough water was added to the batch volume and the solution was filtered through a 0.2 pm PES membrane. An aliquot of the filtered solution was placed in a vial. The vial was sealed with a crimp seal and stored at -20 °C. The batch composition and vial composition are described in Table 31 below.

[1086]

[1087] Table 31: Batch Composition

[1088] Example 53: Analytical Test Method 1

[1089] Reversed phase HPLC using a C8 column: 25 °C, ultraviolet (UV) detection at 225 nm.

[1090] Mobile phase: Gradient system starting with 85% mobile phase A (water with 0.01% trifluoroacetic acid) and 15% mobile phase B (acetonitrile with 0.01% trifluoroacetic acid) and ending with 75% mobile phase B after 40 minutes.

[1091] Samples were prepared by dissolving the contents of a capsule in diluent which was 15:85 (v / v) acetonitrile: 20 mM citrate buffer. Under these aqueous conditions, the compound of formula (1-1) completely hydrolyzes the citrate ester moiety of the molecule to give the compound of formula (VIII-1) in a 1:1 molecular ratio. The presence of the compound of formula (VIII-1) in the sample was confirmed by comparing the sample retention time to the retention time of a reference standard. The amount of compound of formula (VIII-1) present in the sample was calculated from the peak area and the peak area of the reference standard based on a weight comparison (including molecular weight conversion). The reference standard used was a known amount of the compound of formula (1-1) of known purity prepared under the same hydrolysis conditions as the sample. The limit of quantitation for the method was 0.05% and the calculated limit of detection was 0.02%.

[1092] Example 54: Analytical Test Method 2

[1093] Normal phase HPLC using isocratic elution: 40 / 60 / 0.1 (v / v / v) THF / n-hexane / TFA mobile phase on a cyano HPLC column eluted for 8 minutes at 25 °C, UV detection at 230 nm.

[1094] A sample was prepared by dissolving the contents of the capsule in 40 / 60 (v / v) THF / n-hexane. Under the conditions, the compound of formula (I-l) does not hydrolyze to the compound of formula (VIII-l). The amount of compound of formula (VIII-l) present in the sample was calculated based on weight comparison, via the area under the peak and the area under the peak of a standard. The standard used was a known amount of compound of formula (VIII-l) of known purity prepared under the same conditions as the sample. The limit of quantitation for the compound of formula (I-l) was 0.2%.

[1095] To calculate the amount of compound of formula (I-l) present in the sample, both Analytical Test Method 1 and Analytical Test Method 2 were used. Analytical Test Method 1 was used to calculate the amount of compound of formula (VIII-l) by weight present in a sample containing compound of formula (I-l). Analytical Test Method 2 was also used to calculate the amount of compound of formula (VIII-l) present in a sample of compound of formula (I-l) obtained without induced hydrolysis.

[1096] The amount of compound of formula (VIII-l) obtained via Analytical Test Method 1 was subtracted from the amount of compound of formula (VIII-l) obtained via Analytical Test Method 2 to give the measured amount of compound of formula (VIII-l) produced by induced hydrolysis of compound of formula (I-l) present in the sample. Molecular weight calculations were made on a 1 : 1 molecular ratio to give the amount of compound of formula (I-l) present in the sample.

[1097] While the foregoing application has been described in some detail for purposes of clarity and understanding, it will be appreciated that certain changes and modifications can be practiced within the scope of the appended claims and that the particular embodiments described herein are not to be taken as limiting with regard to the application. All literature and scientific articles cited herein are expressly incorporated by reference.

[1098] The patents and scientific articles cited herein are hereby expressly incorporated by reference. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The citation of references herein does not constitute an admission that the references are prior art or that their contents are in any way relevant to the patentability of the application. The disclosure as set forth herein can encompass variations as come within the scope of the claims. Moreover, any one or more features of a claim can be has out in multiple embodiments. Therefore, the application is not limited by the disclosure herein, but instead its scope is to be determined entirely by the following claims, which address and accomplish the features, advantages, and object of the application without regard to their recitation in the summary or description.

Claims

1. A compound, said compound being:

2. A compound, said compound being:

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