Boronate ester compounds and pharmaceutical compositions thereof

Abstract

To provide a boronate ester compound useful as a proteasome inhibitor.SOLUTION: Provided is a compound of Formula (I) or a pharmaceutically acceptable salt thereof. (In the formula, A is 0, 1, or 2, P is hydrogen or an amino-blocking moiety, Ra, Ra1, and Ra2 are hydrogen, C1-6 aliphatic, C1-6 fluoroaliphatic, etc., and Z1 and Z2 together form a moiety derived from an α-hydroxycarboxylic acid, and in any case an atom bonded to boron is an oxygen atom, or Z1 and Z2 together form a moiety derived from a β-hydroxycarboxylic acid).SELECTED DRAWING: None

Description

[Technical Field]

[0001] Priority This application claims priority from 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, both of which are incorporated herein by reference in their entireties.

[0002] The present invention relates to boronic 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 in the treatment of various diseases. [Background technology]

[0003] Boronic acid and boronate ester compounds exhibit various biological activities that are useful for pharmaceutical purposes. U.S. Patent No. 4,499,082 (1985) to Shenvi et al. discloses that peptide boronic acids are inhibitors of certain proteolytic enzymes. U.S. Patent No. 5,187,157 (1993), U.S. Patent No. 5,242,904 (1993), and U.S. Patent No. 5,250,720 (1993) to Kettner and Shenvi describe a class of peptide boronic acids that inhibit trypsin-like proteases. 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 that certain boronic acid compounds inhibit the proliferation 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.

[0004] Boronic acid and ester compounds show particular promise as inhibitors of proteases, the multicatalytic proteinases responsible for most intracellular protein turnover. Adams et al., U.S. Patent No. 5,780,454 (1998), describes peptide boronate ester and acid compounds useful as protease inhibitors. The reference also describes that the use of boronate ester and acid compounds reduces the rate of muscle protein degradation, reduces NF-κB activity in cells, reduces the rate of p53 protein degradation in cells, inhibits cyclin degradation in cells, inhibits cancer cell proliferation, and inhibits 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 additional boronic ester and acid compounds that are reported to have protease inhibitor activity.

[0005] Ciechanover, Cell, 79:13-21 (1994) discloses that the proteasome is a proteolytic component of the ubiquitin-proteasome pathway, in which proteins are targeted for degradation by conjugation with multiple molecules of ubiquitin. Ciechanover also discloses that the ubiquitin-proteasome pathway plays an important role in a variety of important physiological processes. Rivett et al., Biochem. J. 291:1 (1993) discloses that the proteasome exhibits trypsin, chymotrypsin, and peptidylglutamyl peptidase activities. The 20S proteasome constitutes the catalytic core of the 26S proteasome. McCormack et al., Biochemistry 37:7792 (1998) teach that various peptide substrates, including Suc-Leu-Leu-Val-Tyr-AMC, Z-Leu-Leu-Arg-AMC, and Z-Leu-Leu-Glu-2NA (Suc is N-succinyl, AMC is 7-amino-4-methylcoumarin, and 2NA is 2-naphthylamine), are cleaved by the 20S proteasome.

[0006] Proteasome inhibitors represent an important new strategy in cancer therapy. King et al., Science 274:1652-1659 (1996) describe that the ubiquitin-proteasome pathway plays an essential role in regulating the cell cycle, neoplastic growth, and metastasis. The authors report that cyclins and cyclin-dependent kinases p21 and p27 KIP1 They teach that a number of key regulatory proteins, including β-glucanase (β-glucanase), are transiently degraded during the cell cycle by the ubiquitin-proteasome pathway. The ordered degradation of these proteins is necessary for cells to progress through the cell cycle and undergo mitosis.

[0007] Furthermore, the ubiquitin-proteasome pathway is necessary for transcriptional regulation. Palombella et al., Cell, 78:773 (1994) teaches that proteasome-mediated degradation of the inhibitory protein IκB regulates activation of the transcription factor NF-κB. NF-κB, in turn, plays a central role in regulating genes involved in immune and inflammatory responses. Read et al., Immunity, 2:493-506 (1995) teaches that the ubiquitin-proteasome pathway is necessary 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 and extravasation of tumor cells to and from the vasculature to distant tissue sites within the body. Also, Beg and Baltimore, Science 274:782 (1996) teach that NF-κB is an anti-apoptotic factor and that inhibition of NF-κB activation renders cells more sensitive to environmental stresses and cytotoxic drugs.

[0008] The proteasome inhibitor VELCADE® (bortezomib, N-2-pyrazinecarbonyl-L-phenylalanine-L-leucineboronic acid) is the first proteasome inhibitor to achieve regulatory approval. Mitsiades et al., Current Drug Targets, 7:1341 (2006) outline the clinical studies leading to the approval of bortezomib for the treatment of patients with multiple myeloma who had received at least one prior art therapy. Fisher et al., J. Clin. Oncol., 30:4867 (2006) describe an international, multicenter, phase II trial confirming 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 a number of molecular mechanisms that may contribute to the antitumor activity of bortezomib.

[0009] Structural analysis reported by Voges et al., Annu. Rev. Biochem., 68:1015 (1999) indicates that the 20S proteasome contains 28 subunits, with catalytic subunits β1, β2, and β5 responsible for peptidylglutamyl, trypsin, and chymotrypsin peptidase activities, respectively. Rivett et al., Curr. Protein Pept. Sci., 5:15 3 (2004) discloses that when the proteasome is exposed to certain cytokines, including IFN-γ and TNF-α, the β1, β2, and β5 subunits are replaced by alternative catalytic subunits β1i, β2i, and β5i, forming a variant of the proteasome known as the immunoproteasome.

[0010] Orlowski, Hematology (Am. Soc. Hematol. Educ. Program) 220 (2005), also discloses that the immunoproteasome is constitutively expressed in some cells derived from hematopoietic progenitor cells. The authors suggest that immunoproteasome-specific inhibitors could enable targeted therapy for cancers arising from hematopoietic origins, thereby potentially sparing normal tissues such as gastrointestinal and nervous tissues from side effects.

[0011] 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) teaches that arylboronic acid compounds readily form cyclic trimeric anhydrides under dehydrating conditions. Also, alkylboronic acids and their boroxines are often air-sensitive. Korcek et al., J. Chem. Soc., Perkin Trans. 2 242 (1972) teaches that butylboronic acid is readily oxidized by air to produce 1-butanol and boric acid. These problems limit the pharmaceutical usefulness of boronic acid compounds, characterize pharmaceutical agents containing boronic acid compounds, and limit their shelf life.

[0012] US Patent Application Publication No. 2007 / 0129994 to Plamondon et al. discloses stable, pharmaceutically acceptable compounds prepared from boronic acid compounds and sugars. There is a need for more stable formulations of boronic acid compounds. [Prior art documents] [Patent documents]

[0013] [Patent Document 1] International Publication No. 02 / 059131 Summary of the Invention [Means for solving the problem]

[0014] The present invention provides novel boronic ester compounds and stable, pharmaceutically acceptable compositions comprising them that are useful for inhibiting proteasome activity in vitro and in vivo, and are particularly useful for the treatment of various cell proliferative disorders.

[0015] In one aspect, the present invention provides a compound of general formula (I):

[0016] [ka] or a pharmaceutically acceptable salt thereof, wherein: A is 0, 1, or 2; P is hydrogen or an amino group blocking moiety; 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 and 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 and Each R a2 are 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 and Each R B is independently a substituted or unsubstituted monocyclic or bicyclic ring system; Each R 4 are independently hydrogen, or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl group, or two R on the same nitrogen atom 4 together with the nitrogen atom 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 the group consisting of N, O, and S; and each R 5are independently hydrogen or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl group; Each R 5a are independently hydrogen or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl group; Each R 5b are independently hydrogen or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl group; Each R 6 are independently a substituted or unsubstituted aliphatic, aryl, or heteroaryl group; Y is hydrogen, -CN, or -NO2; m is 0, 1, or 2; Z 1 and Z 2 both form a moiety derived from an α-hydroxycarboxylic acid, and in each case the atom attached to the boron is an oxygen atom or Z 1 and Z 2 both form moieties derived from β-hydroxycarboxylic acids, and in both cases the atom attached to the boron is an oxygen atom.

[0017] In another aspect, the present invention provides a pharmaceutical composition comprising a compound of formula (I), or a crystalline form thereof, and additional excipients as described herein, which is suitable for preparing an oral pharmaceutical dosage form.

[0018] In another aspect, the present invention provides a pharmaceutical composition comprising a compound of formula (I), or a crystalline form thereof, and additional excipients as described herein, which is suitable for producing a lyophilized powder pharmaceutical dosage form.

[0019] In another aspect, the present invention provides a pharmaceutical composition comprising a compound of formula (I), or a crystalline form thereof, and additional excipients as described herein, which is suitable for preparing a liquid pharmaceutical dosage form.

[0020] In another aspect, the present invention provides a pharmaceutical composition comprising a compound of formula (I), or a crystalline form thereof, a filler, and optionally a lubricant.

[0021] In another aspect, the present invention provides a pharmaceutical composition comprising a compound of formula (I), or a crystalline form thereof, a filler, optionally a lubricant, optionally a flow aid, and optionally a buffer.

[0022] In another aspect, the present invention provides a pharmaceutical composition comprising a compound of formula (I), or a crystalline form thereof, a bulking agent, and a buffering agent.

[0023] In another aspect, the present invention provides a process for producing the pharmaceutical composition of the present invention.

[0024] In another aspect, the present invention provides methods of using the pharmaceutical compositions of the present invention to treat a patient suffering from or at risk of developing or experiencing a recurrence of a proteasome-mediated disease.

[0025] In another aspect, the present invention provides methods of using the pharmaceutical compositions of the present invention for the treatment of cancer. The present invention provides, for example, the following items. (Item 1) Compounds of formula (I) [ka] or a pharmaceutically acceptable salt thereof, wherein: A is 0, 1, or 2; P is hydrogen or an amino group blocking moiety; 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 and 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 and Each R a2 are 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 and Each R B is independently a substituted or unsubstituted monocyclic or bicyclic ring system; Each R 4 are independently hydrogen, or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl group, or two R on the same nitrogen atom 4 together with the nitrogen atom form a substituted or unsubstituted 4- to 8-membered heterocyclyl ring which, in addition to the nitrogen atom, has 0 to 2 ring heteroatoms independently selected from N, O, and S; Each R 5 are independently hydrogen or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl group; Each R 5a are independently hydrogen or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl group; Each R 5b are independently hydrogen or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl group; Each R 6 are independently a substituted or unsubstituted aliphatic, aryl, or heteroaryl group; Y is hydrogen, -CN, or -NO2; m is 0, 1, or 2; Z 1 and Z 2 both form a moiety derived from an α-hydroxycarboxylic acid, and in each case the atom bonded to the boron is an oxygen atom or Z 1 and Z 2 and R 1 and R 2 together form a moiety derived from a β-hydroxycarboxylic acid, and in each case the atom bonded to the boron is an oxygen atom, or a pharmaceutically acceptable salt thereof. (Item 2) Formula (II) [ka] wherein: R b1 and R b2 each is independently hydrogen, —COH, —OH, or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl group; R b3 and R b4 each is independently hydrogen, —COH, or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl group; Or, R b2 and R b4 are each independently hydrogen, and R b1 and R b3 together with the carbon atoms to which they are attached form an unsubstituted or substituted fused 4-8 membered non-aromatic ring having 0-3 ring heteroatoms selected from the group consisting of O, N, and S, wherein said ring can be optionally fused to an unsubstituted or substituted 4-8 membered non-aromatic ring or a 5-6 membered aromatic ring having 0-3 ring heteroatoms selected from the group consisting of O, N, and S; or R b2 and R b4 does not exist, R b1 and R b3 together with the carbon atoms to which they are attached form an unsubstituted or substituted fused 5-6 membered aromatic ring having 0-3 ring heteroatoms selected from the group consisting of O, N, and S, wherein said ring can be optionally fused to an unsubstituted or substituted 4-8 membered non-aromatic ring or a 5-6 membered aromatic ring having 0-3 ring heteroatoms selected from the group consisting of O, N, and S; The compound according to item 1, wherein n is 0 or 1, or a pharmaceutically acceptable salt thereof. (Item 3) R b3 and R b4 each independently represents hydrogen, C 1-6 Aliphatic, or -(CH2) p -CO2H, 3. The compound according to item 2, wherein p is 0, 1, or 2. (Item 4) 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-, R c is C 1-6 aliphatic, C 1-6 Fluoroaliphatic, -R D , -T 1 -R D , and -T 1 -R 2c is selected from the group consisting of T 1 is 0 to 2 independently selected R 3a or R 3b Replaced by C 1-6 an alkylene chain, the alkylene chain optionally being —C(R 5 )=C(R 5 )-, -C≡C-, or -O- interrupted, R D is a substituted or unsubstituted monocyclic or bicyclic ring system, R 2c , 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(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)-(R 4 )2, -C(O)-R5 , -CO2R 5 , or -C(O)N(R 4 )2, Each R 3a are independently -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 are independent, R 3a or R 7 Substituted or unsubstituted C 1-3 aliphatic or two substituents R on the same carbon atom 3b together with the carbon atoms to which they are attached form a 3- to 6-membered cycloaliphatic ring, Each R 7 is a substituted or unsubstituted aromatic group, R 4c is hydrogen, C 1-4 Alkyl, C 1-4 Fluoroalkyl, or C 6-10 Ara (C 1-4 ) alkyl, and the aryl portion thereof is substituted or unsubstituted. (Item 5) A is 0, R a1 is hydrogen, -(CH2) m -CH2-R B , or -(CH2) m -CH(R 5a )-OR 5b and 4. The compound according to item 3, wherein m is 0. (Item 6) P is R c -C(O)- or R c -S(O)2- and R c -R D Item 4. The compound according to item 3, wherein (Item 7) R Dis a substituted or unsubstituted monocyclic or bicyclic ring system selected from the group consisting of phenyl, pyridinyl, pyrimidinyl, pyrazinyl, naphthyl, benzimidazolyl, quinolinyl, isoquinolinyl, quinoxalinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, tetrahydroquinoxalinyl, and dihydrobenzoxazinyl. (Item 8) R D is 0 to 1 R d and 0 to 2 R 8d is substituted on a substitutable carbon atom with Each R d independently, C 1-6 aliphatic, C 1-6 fluoroaliphatic, or halo; Each R 8d independently, C 1-4 aliphatic, C 1-4 8. The compound according to item 7, which is fluoroaliphatic or halo. (Item 9) A is 0, R a is C 1-6 Aliphatic, or -(CH2) m -CH2-R B and R a1 is hydrogen, -(CH2) m -CH2-R B , or -(CH2) m -CH(R 5a )-OR 5b and P is R c -C(O)- or R c -S(O)2-, R c -R D and 5. The compound according to item 4, wherein m is 0 or 1. (Item 10) R a is C 1-6 10. The compound according to item 9, which is aliphatic. (Item 11) R a1 is -CH2-R B and R B is phenyl, R D Item 11. The compound according to item 10, wherein is 2-pyrazinyl. (Item 12) R a1 is -CH(R 5a )-OR 5b and R 5a is C 1-6 is aliphatic, R 5b is hydrogen, R D 11. The compound according to item 10, wherein is 6-phenyl-2-pyridinyl-. (Item 13) R a1 is hydrogen, R D Item 11. The compound according to item 10, wherein is 2,5-dichlorophenyl. (Item 14) Z 1 and Z 2 and together form a moiety derived from citric acid. (Item 15) The compound according to item 14, which is in a substantially crystalline form. (Item 16) Formulas (III), (IIIa), (IV), (IVa) [ka] [ka] Item 15. The compound according to item 14, characterized in that: (Item 17) R a is C 1-6 17. The compound according to item 16, which is aliphatic. (Item 18) A is 0 and R a1 is hydrogen, -(CH2) m -CH2-R B , or -(CH2) m -CH(R 5 )-OR 5b and P is R c -C(O)- and R c -R D Item 17. The compound according to item 16, wherein (Item 19) R D is a substituted or unsubstituted monocyclic or bicyclic ring system selected from the group consisting of phenyl, pyridinyl, pyrimidinyl, pyrazinyl, naphthyl, benzimidazolyl, quinolinyl, isoquinolinyl, quinoxalinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, tetrahydroquinoxalinyl, and dihydrobenzoxazinyl. (Item 20) R a Item 19. The compound according to item 18, wherein is isobutyl. (Item 21) R a1 is -CH2-R B and R B is phenyl, R D 21. The compound according to item 20, wherein is 2-pyrazinyl. (Item 22) R a1 is hydrogen, R D 21. The compound according to item 20, wherein is 2,5-dichlorophenyl. (Item 23) R a1 is -CH(R 5a )-OR 5b and R 5a is C 1-6 is aliphatic, R 5b is hydrogen, R D 21. The compound according to item 20, wherein is 6-phenyl-2-pyridinyl-. (Item 24) Compound of formula (I) [ka] or a crystalline form thereof, a filler, and optionally a lubricant, During the ceremony, A is 0, 1, or 2; P is hydrogen or an amino group blocking moiety; 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 and 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 and Each R a2 are 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(R4 )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 and Each R B is independently a substituted or unsubstituted monocyclic or bicyclic ring system; Each R 4 are independently hydrogen, or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl group, or two R on the same nitrogen atom 4 together with the nitrogen atom form a substituted or unsubstituted 4- to 8-membered heterocyclyl ring which, in addition to the nitrogen atom, has 0 to 2 ring heteroatoms independently selected from N, O, and S; Each R 5 are independently hydrogen or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl group; Each R 5a are independently hydrogen or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl group; Each R 5b are independently hydrogen or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl group; Each R 6 are independently a substituted or unsubstituted aliphatic, aryl, or heteroaryl group; Y is hydrogen, -CN, or -NO2; m is 0, 1, or 2; Z 1 and Z 2 both form a moiety derived from an α-hydroxycarboxylic acid, and in each case the atom attached to the boron is an oxygen atom or Z 1and Z 2 and (b) form a moiety derived from a β-hydroxycarboxylic acid, and in each case the atom attached to the boron is an oxygen atom. (Item 25) The pharmaceutical composition according to Item 24, optionally further comprising a flow aid and optionally further comprising a buffer. (Item 26) The pharmaceutical composition comprises, as a percentage of the total weight, about 0.2% by weight to about 3% by weight of the compound of formula (I) or a crystalline form thereof, about 86.5% by weight to about 99.8% by weight of a filler 26. The pharmaceutical composition of item 25, optionally comprising up to about 1.5% by weight of a lubricant, optionally up to about 5% by weight of a flow aid, and optionally up to about 5% by weight of a buffer. (Item 27) The pharmaceutical composition according to Item 25, wherein the pharmaceutical composition comprises, as a percentage of total weight, about 0.2% to about 3% by weight of the compound of formula (I) or a crystalline form thereof, about 97% to about 99.8% by weight of a filler, and optionally up to about 1.5% by weight of a lubricant. (Item 28) The pharmaceutical composition according to Item 25, wherein the pharmaceutical composition comprises, as a percentage of total weight, about 0.25% by weight to about 2% by weight of the compound of formula (I) or a crystalline form thereof, and about 98% by weight to about 99.75% by weight of a filler. (Item 29) The pharmaceutical composition according to Item 25, wherein the pharmaceutical composition is an oral pharmaceutical dosage form. (Item 30) The pharmaceutical composition according to Item 29, wherein the oral pharmaceutical dosage form is a capsule. (Item 31) The pharmaceutical composition according to 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 as a percentage of the total weight. (Item 32) The pharmaceutical composition according to Item 25, wherein the compound of formula (I), or a crystalline form thereof, is present in an amount of about 0.25% by weight to about 2% by weight as a percentage of the total weight. (Item 33) The pharmaceutical composition according to Item 25, wherein the filler is present in an amount of about 97% to about 99.8% by weight as a percentage of the total weight. (Item 34) The pharmaceutical composition according to Item 25, wherein the filler is present in an amount of about 98% by weight to about 99.75% by weight as a percentage of the total weight. (Item 35) The pharmaceutical composition according to Item 25, wherein the filler is present in an amount of about 86.5% by weight to about 99.8% by weight as a percentage of the total weight. (Item 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. (Item 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. (Item 38) The pharmaceutical composition of Item 25, wherein the lubricant, if present, is present in an amount of up to about 1.5% by weight as a percentage of the total weight. (Item 39) The pharmaceutical composition according to Item 25, wherein the lubricant, if present, is present in an amount of about 1% by weight as a percentage of the total weight. (Item 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 stearate fumarate, and mixtures thereof. (Item 41) The pharmaceutical composition according to Item 25, wherein the lubricant is magnesium stearate. (Item 42) The pharmaceutical composition of Item 25, wherein the flow aid, when present, is present in an amount of up to about 5% by weight as a percentage of the total weight. (Item 43) The pharmaceutical composition of Item 25, wherein the flow aid, if present, is present in an amount of up to about 1% by weight as a percentage of the total weight. (Item 44) The pharmaceutical composition of Item 25, wherein the flow aid, if present, is talc. (Item 45) The pharmaceutical composition according to Item 25, wherein the buffering agent, if present, is present in an amount of up to about 5% by weight as a percentage of the total weight. (Item 46) The pharmaceutical composition according to Item 25, wherein the buffering agent, if present, is present in an amount of up to about 2% by weight as a percentage of the total weight. (Item 47) The method according to Item 25, wherein the buffer, if present, is sodium citrate. The pharmaceutical composition described above. (Item 48) The α-hydroxycarboxylic acid or β-hydroxycarboxylic acid is citric acid, 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 and P is R c -C(O)-, R c -R D and m is 0 or 1, the filler is selected from the group consisting of low moisture microcrystalline cellulose, sodium starch glycolate, pregelatinized starch, and mixtures thereof; 26. The pharmaceutical composition according to item 25, wherein the lubricant, if present, is magnesium stearate. (Item 49) The compound of formula (I) is represented by compound (I-1), (I-15), or (I-18), [ka] [ka] the filler is selected from the group consisting of low moisture microcrystalline cellulose, sodium starch glycolate, pregelatinized starch, and mixtures thereof; 26. The pharmaceutical composition according to item 25, wherein the lubricant, if present, is magnesium stearate. (Item 50) The α-hydroxycarboxylic acid or β-hydroxycarboxylic acid is citric acid; 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 and P is R c -C(O)-, R c -R D and m is 0 or 1, the filler is selected from the group consisting of low moisture microcrystalline cellulose, sodium starch glycolate, pregelatinized starch, and mixtures thereof; the lubricant, if present, is magnesium stearate; the flow aid, if present, is talc; 26. The pharmaceutical composition of item 25, wherein the buffering agent, if present, is sodium citrate. (Item 51) The compound of formula (I) is represented by compound (I-1), (I-15), or (I-18), [ka] the filler is selected from the group consisting of low moisture microcrystalline cellulose, sodium starch glycolate, pregelatinized starch, and mixtures thereof; the lubricant, if present, is magnesium stearate; the flow aid, if present, is talc; 26. The pharmaceutical composition of item 25, wherein the buffering agent, if present, is sodium citrate. (Item 52) Compound of formula (I-1) [ka] or a crystalline form thereof, a filler, and optionally a lubricant. 53. The filler is selected from the group consisting of low-moisture microcrystalline cellulose, sodium starch glycolate, pregelatinized starch, and mixtures thereof; 53. The pharmaceutical composition of item 52, wherein the lubricant, if present, is magnesium stearate. (Item 54) The pharmaceutical composition according to Item 52, wherein the crystalline form is Form 2. (Item 55) The pharmaceutical composition according to Item 52, optionally further comprising a flow aid and optionally further comprising a buffer. 56. The filler is selected from the group consisting of low-moisture microcrystalline cellulose, sodium starch glycolate, pregelatinized starch, and mixtures thereof; the lubricant, if present, is magnesium stearate; the flow aid, if present, is talc; 56. The pharmaceutical composition of item 55, wherein the buffering agent, if present, is sodium citrate. (Item 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 equivalent to about 0.1 mg to about 3.0 mg of the compound of formula (VIII-1) on a molar weight basis. (Item 58) A unit dose pharmaceutical composition comprising about 0.143 mg to about 4.3 mg of the 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), on a weight / weight basis. (Item 59) Compound of formula (I) [ka] or a crystalline form thereof, a bulking agent, and a buffering agent, During the ceremony, A is 0, 1, or 2; P is hydrogen or an amino group blocking moiety; Ra 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 and 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 ,Ma TAHA-(CH2) m -CH(R 5 )-SR 5 and Each R a2 are 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 and Each R B is independently a substituted or unsubstituted monocyclic or bicyclic ring system; Each R 4 are independently hydrogen, or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl group, or two R on the same nitrogen atom 4 together with the nitrogen atom form a substituted or unsubstituted 4- to 8-membered heterocyclyl ring which, in addition to the nitrogen atom, has 0 to 2 ring heteroatoms independently selected from N, O, and S; Each R 5 are independently hydrogen or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl group; Each R 5a are independently hydrogen or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl group; Each R 5b are independently hydrogen or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl group; Each R 6 are independently a substituted or unsubstituted aliphatic, aryl, or heteroaryl group; Y is hydrogen, -CN, or -NO2; m is 0, 1, or 2; Z 1 and Z 2both form a moiety derived from an α-hydroxycarboxylic acid, and in each case the atom attached to the boron is an oxygen atom or Z 1 and Z 2 and (b) form a moiety derived from a β-hydroxycarboxylic acid, and in each case the atom attached to the boron is an oxygen atom. (Item 60) The pharmaceutical composition according to Item 59, wherein the bulking agent is present in an amount of about 1% w / v to about 5% w / v. (Item 61) The pharmaceutical composition of Item 59, wherein the bulking agent is present in an amount of about 3% w / v. (Item 62) The pharmaceutical composition of Item 59, wherein the bulking agent is glycine. (Item 63) The pharmaceutical composition of Item 59, wherein the buffering agent is sodium citrate and citric acid. (Item 64) The α-hydroxycarboxylic acid or β-hydroxycarboxylic acid is citric acid, 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 and P is R c -C(O)-, R c -R D and m is 0 or 1, the bulking agent is glycine; 60. The pharmaceutical composition of claim 59, wherein the buffering agent is sodium citrate and citric acid. (Item 65) The compound of formula (I) is represented by compound (I-1), (I-15), or (I-18), [ka] the bulking agent is glycine; 60. The pharmaceutical composition of claim 59, wherein the buffering agent is sodium citrate and citric acid. (Item 66) A pharmaceutical composition comprising a lyophilized powder of a compound of formula (I), [ka] During the ceremony, A is 0, 1, or 2; P is hydrogen or an amino group blocking moiety; 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 and 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 ,Ma TAHA-(CH2) m -CH(R 5 )-SR 5 and Each R a2 are 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 and Each R B is independently a substituted or unsubstituted monocyclic or bicyclic ring system; Each R 4 are independently hydrogen, or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl group, or two R on the same nitrogen atom 4 together with the nitrogen atom form a substituted or unsubstituted 4- to 8-membered heterocyclyl ring which, in addition to the nitrogen atom, has 0 to 2 ring heteroatoms independently selected from N, O, and S; Each R 5 are independently hydrogen or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl group; Each R 5a are independently hydrogen or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl group; Each R 5b are independently hydrogen or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl group; Each R 6 are independently a substituted or unsubstituted aliphatic, aryl, or heteroaryl group; Y is hydrogen, -CN, or -NO2; m is 0, 1, or 2; Z 1 and Z 2 both form a moiety derived from an α-hydroxycarboxylic acid, and in each case the atom attached to the boron is an oxygen atom or Z 1 and Z 2 and (b) form a moiety derived from a β-hydroxycarboxylic acid, and in each case the atom attached to the boron is an oxygen atom. (Item 67) The pharmaceutical composition according to Item 66, further comprising a bulking agent and a buffering agent. (Item 68) The pharmaceutical composition of Item 67, wherein the bulking agent is glycine. (Item 69) The pharmaceutical composition according to Item 67, wherein the buffering agent is sodium citrate and citric acid. (Item 70) The α-hydroxycarboxylic acid or β-hydroxycarboxylic acid is citric acid; 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 and P is R c -C(O)-, R c -R D and m is 0 or 1, the bulking agent is glycine; 68. The pharmaceutical composition of claim 67, wherein the buffering agent is sodium citrate and citric acid. (Item 71) The compound of formula (I) is represented by compound (I-1), (I-15), or (I-18), [ka] the bulking agent is glycine; 68. The pharmaceutical composition of claim 67, wherein the buffering agent is sodium citrate and citric acid. (Item 72) A unit dose pharmaceutical composition comprising a lyophilized powder of the compound of formula (I-1), [ka] During the ceremony, A unit dose pharmaceutical composition, wherein the compound of formula (I-1) is present in an amount equivalent to about 1 mg to about 5 mg of the compound of formula (VIII-1) on a molar weight basis. (Item 73) The unit dose pharmaceutical composition according to Item 72, wherein the compound of formula (I-1) is present in an amount equivalent to about 3.5 mg of the compound of formula (VIII-1) on a molar weight basis. (Item 74) The unit dose pharmaceutical composition of Item 72, further comprising glycine, sodium citrate, and citric acid. (Item 75) The amount of glycine is about 0.01 g to about 0.50 g. A unit dose pharmaceutical composition according to claim 1. (Item 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. (Item 77) A unit dose pharmaceutical composition comprising a lyophilized powder of the compound of formula (I-15), [ka] During the ceremony, A unit dose pharmaceutical composition, wherein 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) on a molar weight basis. (Item 78) The unit dose pharmaceutical composition according to 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) on a molar weight basis. (Item 79) The unit dose pharmaceutical composition of Item 78, further comprising glycine, sodium citrate, and citric acid. (Item 80) The unit dose pharmaceutical composition of Item 78, wherein the amount of glycine is present in an amount of about 0.01 g to about 0.50 g. (Item 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. (Item 82) A method for preparing a pharmaceutical composition of compound (I-1) as a lyophilized powder, comprising: (f-1) i. Water, ii. the compound (I-1), iii. glycine, iv. sodium citrate, and v. citric acid to form a mixture; (f-2) freeze-drying the mixture. (Item 83) A method for preparing a pharmaceutical composition of compound (I-15) as a lyophilized powder, comprising: (g-1) i. an aqueous solvent mixture comprising water and tert-butyl alcohol; ii. The compound (VIII-15), iii. glycine, iv. sodium citrate, and v. citric acid to form a mixture; (g-2) freeze-drying the mixture. (Item 84) The method according to Item 83, wherein the tert-butyl alcohol in the aqueous solvent mixture is present in an amount of about 3% v / v to about 6% v / v of tert-butyl alcohol. (Item 85) A method for preparing a pharmaceutical composition of compound (I-1) as a liquid pharmaceutical dosage form, the process comprising reconstituting a lyophilized powder of compound (I-1) according to Item 72 with water for injection. (Item 86) For preparing a pharmaceutical composition of compound (I-15) as a liquid pharmaceutical dosage form, 78. A method for the preparation of a lyophilized powder of compound (I-15) according to item 77, comprising reconstituting the lyophilized powder of compound (I-15) according to item 77 with water for injection. (Item 87) Compound of formula (I) [ka] 1. A liquid pharmaceutical composition comprising a buffer, and optionally a tonicity adjusting agent, During the ceremony, A is 0, 1, or 2; P is hydrogen or an amino group blocking moiety; 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 and R a1 is hydrogen, C 1-6aliphatic, 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 and Each R a2 are 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 and Each R B is independently a substituted or unsubstituted monocyclic or bicyclic ring system; Each R 4 are independently hydrogen, or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl group, or two R on the same nitrogen atom4 together with the nitrogen atom form a substituted or unsubstituted 4- to 8-membered heterocyclyl ring which, in addition to the nitrogen atom, has 0 to 2 ring heteroatoms independently selected from N, O, and S; Each R 5 are independently hydrogen or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl group; Each R 5a are independently hydrogen or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl group; Each R 5b are independently hydrogen or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl group; Each R 6 are independently a substituted or unsubstituted aliphatic, aryl, or heteroaryl group; Y is hydrogen, -CN, or -NO2; m is 0, 1, or 2; Z 1 and Z 2 both form a moiety derived from an α-hydroxycarboxylic acid, and in each case the atom attached to the boron is an oxygen atom or Z 1 and Z 2 and (b) form a moiety derived from a β-hydroxycarboxylic acid, and in each case the atom attached to the boron is an oxygen atom. (Item 88) The tonicity modifier, if present, 88. The liquid pharmaceutical composition according to item 87, wherein the active ingredient is sodium chloride. (Item 89) The liquid pharmaceutical composition according to Item 87, wherein the buffer is sodium citrate and citric acid. (Item 90) The α-hydroxycarboxylic acid or β-hydroxycarboxylic acid is citric acid, A is 0, R a is isobutyl, R a1 is hydrogen, C 1-6Aliphatic, -(CH2) m -CH2-R B , or -(CH2) m -CH(R 5a )-OR 5b and P is R c -C(O)-, R c -R D and m is 0 or 1, the buffering agent is sodium citrate and citric acid; 88. The liquid pharmaceutical composition of item 87, wherein the tonicity adjusting agent, if present, is sodium chloride. (Item 91) A unit dose liquid pharmaceutical composition comprising a compound of formula (I-1), [ka] During the ceremony, A unit dose liquid pharmaceutical composition, wherein the compound of formula (I-1) is present in a concentration of the compound of formula (VIII-1) from about 0.5 mg / mL to about 3 mg / mL. (Item 92) The unit-dose liquid pharmaceutical composition of Item 91, further comprising sodium citrate, citric acid, and sodium chloride. (Item 93) A method for treating cancer, comprising administering a therapeutically effective amount of the pharmaceutical composition described in any one of Items 24, 52, 57, 58, 59, 72, 77, and 87. [Brief explanation of the drawings]

[0026] [Figure 1] FIG. 1 is a powder X-ray diffraction diagram of 4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-methylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid (I-1) Form 1. [Figure 2]FIG. 1 is a differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) profile of 4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-methylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid (I-1) Form 1. [Figure 3] Powder X-ray diffraction of 4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-methylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid (I-1) Form 2. [Figure 4] FIG. 1 is a differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) profile of 4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-methylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid (I-1) Form 2. [Figure 5] Powder X-ray diffraction 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). [Figure 6] Powder X-ray diffraction of 2,5-dichloro-N-(2-{[(1R)-3-dichloro-1-(4-oxo-4H-1,3,2-benzodioxaborinin-2-yl)butyl]amino}-2-oxoethyl)benzamide (I-13). [Figure 7] Powder X-ray diffraction of 4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-methylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid (I-1) Form 2. [Figure 8] FIG. 1 is a differential scanning calorimetry (DSC) profile of 4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-methylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid (I-1) Form 2. DETAILED DESCRIPTION OF THE INVENTION

[0027] definition Unless expressly stated otherwise, the term "proteasome" is intended to refer to both constitutive and immunoproteasomes.

[0028] The term "aliphatic" or "aliphatic group," as used herein, refers to a substituted or unsubstituted straight-chain, branched-chain, or cyclic C 1-12 "Aliphatic" refers to a hydrocarbon that is either fully saturated or contains one or more units of unsaturation, but is not aromatic. For example, suitable aliphatic groups include substituted or unsubstituted linear, branched, or cyclic alkyl, alkenyl, or alkynyl groups, such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl, or (cycloalkyl)alkenyl, and mixtures thereof. In various embodiments, the aliphatic group has 1 to 12, 1 to 8, 1 to 6, 1 to 4, or 1 to 3 carbons.

[0029] The terms "alkyl," "alkenyl," and "alkynyl," used alone or as part of a larger moiety, refer to straight- or branched-chain aliphatic groups having 1 to 12 carbon atoms. For purposes of the present invention, the term "alkyl" may be used when the carbon atom attaching the aliphatic group to the rest of the molecule is a saturated carbon atom. However, alkyl groups may contain unsaturation at other carbon atoms. Thus, alkyl groups include, but are not limited to, methyl, ethyl, propyl, allyl, propargyl, butyl, pentyl, and hexyl.

[0030] For purposes of the present invention, the term "alkenyl" may be used when the carbon atom attaching the aliphatic group to the rest of the molecule forms part of a carbon-carbon double bond. Alkenyl groups include, but are not limited to, vinyl, 1-propenyl, 1-butenyl, 1-pentenyl, and 1-hexenyl.

[0031] For purposes of the present invention, the term "alkynyl" may be used when the carbon atom attaching the aliphatic group to the rest of the molecule forms 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.

[0032] The term "cycloalkyl," used alone or as part of a larger moiety, refers to a saturated or partially unsaturated cyclic aliphatic ring system having 3 to about 14 members, which aliphatic ring systems are optionally substituted. In some embodiments, the cycloaliphatic is a monocyclic hydrocarbon having 3 to 8 or 3 to 6 ring carbon atoms. Non-limiting examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclopropyl ... Examples include cyclohexenyl, cycloheptyl, cycloheptenyl, cyclooctyl, cyclooctenyl, and cyclooctadienyl. In some embodiments, the cycloaliphatic is a bridged or fused bicyclic ring having 6 to 12, 6 to 10, or 6 to 8 ring carbon atoms, and any individual ring in the bicyclic ring system has 3 to 8 members.

[0033] In some embodiments, two adjacent substituents on a cycloaliphatic ring, together with the intervening ring atoms, form an optionally substituted fused 5-6-membered aromatic or 3-8-membered non-aromatic ring having 0-3 ring heteroatoms selected from the group consisting of O, N, and S. Thus, the term "cycloaliphatic" includes an aliphatic ring fused to one or more aryl, heteroaryl, or heterocyclyl rings. Non-limiting examples include indanyl, 5,6,7,8-tetrahydroquinoxalinyl, decahydronaphthyl, or tetrahydronaphthyl, where the radical or point of attachment is on the aliphatic ring.

[0034] The terms "aryl" and "ar-", used alone or as part of a larger moiety (e.g., "aralkyl," "aralkoxy," or "aryloxyalkyl"), refer to alkyl groups containing 1 to 3 rings, C 6-14 Aryl groups refer to aromatic hydrocarbons, each of which is optionally substituted. Preferably, an aryl group is C 6-10An aryl group. Aryl groups include, but are not limited to, phenyl, naphthyl, and anthracenyl. In some embodiments, two adjacent substituents on an aryl ring, together with the intervening ring atoms, form an optionally substituted fused 5-6-membered aromatic or 4-8-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, cycloaliphatic, or heterocyclyl rings, where the radical or point of attachment is on the aromatic ring. Non-limiting examples of such fused ring systems include indolyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzthiazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, carbazonyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, fluorenyl, indanyl, phenanthridinyl, tetrahydronaphthyl, indolinyl, phenoxazinyl, benzodioxanyl, and benzodioxolyl. Aryl groups can be monocyclic, bicyclic, tricyclic, or polycyclic, preferably monocyclic, bicyclic, or tricyclic, more preferably monocyclic or bicyclic. The term "aryl" can be used interchangeably with the terms "aryl group," "aryl moiety," and "aryl ring."

[0035] An "aralkyl" or "arylalkyl" group includes an aryl group covalently bonded to an alkyl group, either of which independently is optionally substituted. Preferably, the aralkyl group is 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.

[0036] The terms "heteroaryl" and "heteroar-," used alone or as part of a larger moiety (e.g., heteroaralkyl or "heteroaralkoxy"), refer to groups having 5 to 14 ring atoms, preferably 5, 6, 9, or 10 ring atoms, groups having 6, 10, or 14 pi electrons shared in a cyclic array, and groups having 1 to 4 heteroatoms in addition to carbon atoms. The term "heteroatom" refers to nitrogen, oxygen, or sulfur, and includes any oxygen atom of nitrogen or sulfur.

[0023] The term "nitrogen" includes oxidized forms, and any quaternized form of a basic nitrogen. Thus, when used in reference to a heteroaryl ring atom, the term "nitrogen" includes oxidized nitrogen (as in pyridine N-oxide). Certain nitrogen atoms of five-membered heteroaryl groups can also be substituted, as further defined below. Heteroaryl groups include, but are not limited to, radicals derived from thiophene, furan, pyrrole, imidazole, pyrazole, triazole, tetrazole, oxazole, isoxazole, oxadiazole, thiazole, isothiazole, thiadiazole, pyridine, pyridazine, pyrimidine, pyrazine, indolizine, naphthyridine, pteridine, pyrrolopyridine, imidazopyridine, oxazolopyridine, thiazolopyridine, triazolopyridine, pyrrolopyrimidine, purine, and triazolopyrimidine. As used herein, the phrase "radical derived from" refers to a monovalent radical formed by removing a hydrogen radical from a parent heteroaromatic ring system. The radical (i.e., the point of attachment of the heteroaryl to the rest of the molecule) can be formed at any substitutable position on any ring of the parent heteroaryl ring system.

[0037] In some embodiments, two adjacent substituents on a heteroaryl, together with the intervening ring atoms, form an optionally substituted fused 5-6 membered aromatic or 4-8 membered non-aromatic ring having 0-3 ring heteroatoms selected from the group consisting of O, N, and S. Thus, the terms "heteroaryl" and "heteroa-," as used herein, also include groups in which a heteroaromatic ring is fused to one or more aryl, cycloaliphatic, or heterocyclyl rings, where the radical or point of attachment is on the heteroaromatic ring. Non-limiting examples include indolyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzthiazolyl, benzoxazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazonyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and pyrido[2,3-b]-1,4-oxazin-3(4H)-one. Heteroaryl groups can be monocyclic, bicyclic, tricyclic, or polycyclic, preferably monocyclic, bicyclic, or tricyclic, more preferably monocyclic or bicyclic. The term "heteroaryl" can be used interchangeably with the terms "heteroaryl ring" or "heteroaryl group," each of which includes rings that are optionally substituted. The term "heteroaralkyl" refers to an alkyl group substituted by a heteroaryl, where the alkyl and heteroaryl portions independently are optionally substituted.

[0038] As used herein, the terms "aromatic ring" and "aromatic ring system" refer to an optionally substituted monocyclic, bicyclic, or tricyclic group having 0 to 6, preferably 0 to 4, ring heteroatoms and 6, 10, or 14 pi-electrons shared in a cyclic array. Thus, the terms "aromatic ring" and "aromatic ring system" encompass both aryl and heteroaryl groups.

[0039] As used herein, the terms "heterocycle," "heterocyclyl," "heterocyclic radical," and "heterocycle" are used interchangeably and refer to a stable 3- to 7-membered monocyclic moiety, or a fused 7- to 10-membered ring, or a bridged 6- to 10-membered bicyclic heterocyclic moiety, which is saturated or partially unsaturated and has, in addition to carbon atoms, one or more, preferably one to four, heteroatoms (as described above). When used in reference to a ring atom of a heterocycle, the term "nitrogen" includes substituted nitrogen. By way of example, in a heterocyclyl ring having one to three heteroatoms selected from the group consisting of oxygen, sulfur, or nitrogen, the nitrogen can be N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl), or + It can be NR (such as in the case of N-substituted pyrrolidinyl). A heterocycle can be attached to its pendant group at any heteroatom or carbon atom that results in a stable structure, and any ring atom can be optionally substituted. Such saturated or Examples of partially unsaturated heterocyclic radicals include, but are not limited to, tetrahydrofuranyl, tetrahydrothienyl, pyrrolidinyl, pyrrolidonyl, piperidinyl, pyrrolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and quinuclidinyl.

[0040] In some embodiments, two adjacent substituents on a heterocycle, together with the intervening ring atoms, form an optionally substituted fused 5-6-membered aromatic or 3-8-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," and "heterocyclic radical" can be used interchangeably herein and include groups in which a heterocyclyl ring is fused to one or more aryl, heteroaryl, or cycloaliphatic rings, such as indolyl, 3H-indolyl, chromanyl, phenanthridinyl, or tetrahydroquinolinyl, where the radical or point of attachment is on the heterocyclyl ring. Heterocyclyl groups 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 by a heterocyclyl, wherein the alkyl and heterocyclyl portions independently are optionally substituted.

[0041] As used herein, the term "partially unsaturated" refers to a ring moiety that includes at least one double or triple bond between ring atoms. As defined herein, the term "partially unsaturated" is intended to encompass rings with multiple sites of unsaturation, but is not intended to include aryl or heteroaryl moieties.

[0042] The terms "haloaliphatic," "haloalkyl," "haloalkenyl," and "haloalkoxy" refer to an aliphatic, alkyl, alkenyl, or alkoxy group, optionally substituted with one or more halogen atoms. As used herein, the term "halogen" or "halo" refers to F, Cl, Br, or I. The term "fluoroaliphatic" refers to a haloaliphatic group, where the halogen is fluoro, including perfluoroaliphatic 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.

[0043] The term "linker group" or "linker" refers to an organic moiety that connects two parts of a compound. Linkers generally include atoms such as oxygen or sulfur, units such as -NH-, -CH2-, -C(O)-, -C(O)NH-, or chains of atoms such as alkylene chains. The molecular weight of a linker is generally in the range of about 14 to 200, preferably about 14 to 96, with a length of up to about 6 atoms. In some embodiments, a linker is a C 1-6 It is an alkylene chain.

[0044] The term "alkylene" refers to a divalent alkyl group. An "alkylene chain" is a polymethylene group, i.e., -(CH2) y -, where y is a natural number, preferably 1 to 6, 1 to 4, 1 to 3, 1 to 2, or 2 to 3. A substituted alkylene chain is a polymethylene group in which one or more methylene hydrogen atoms are replaced with a substituent. Suitable substituents include those described below for a substituted aliphatic group. The alkylene chain may also be substituted at one or more positions with an aliphatic group or a substituted aliphatic group.

[0045] An alkylene chain can also be optionally interrupted by a functional group. An alkylene chain is "interrupted" by a functional group when an internal methylene unit is substituted with a functional group. Examples of "functional groups that can be used" 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 )=NO-, or -N(R + )-N(R + )-. Each R + are independently hydrogen or an optionally substituted aliphatic, aryl, heteroaryl, or heterocyclyl group, or two R on the same nitrogen atom + together with the nitrogen atom form a 5-8 membered aromatic or non-aromatic ring having, in addition to the nitrogen atom, 0-2 ring heteroatoms selected from the group consisting of N, O, and S. Each R * are independently hydrogen or an optionally substituted aliphatic, aryl, heteroaryl, or heterocyclyl group. Each Ro is independently an optionally substituted aliphatic, The group is an aromatic, aryl, or heteroaryl group.

[0046] C "divided" by -O- 3-6 Examples of alkylene chains include CHOCH-, -CHO(CH)-, -CHO(CH)-, -CHO(CH)-, -(CH)OCH-, -(CH)O(CH)-, -(CH)O(CH)-, -(CH)O(CH)-, -(CH)O(CH)-, -(CH)O(CH)-, -(CH)O(CH)-, and -(CH)O(CH)-. Other examples of alkylene chains "interrupted" by functional groups include -CHZ * 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 "interrupted" functional groups described above.

[0047] Those skilled in the art will recognize that when an alkylene chain having a break is attached to a functional group, certain combinations may not be sufficiently stable for pharmaceutical use. Only stable or chemically feasible compounds are within the scope of the present invention. A stable or chemically feasible compound is one that maintains its integrity long enough to be useful for therapeutic or prophylactic administration to a patient. Preferably, the chemical structure remains substantially unchanged when maintained at temperatures below -70°C, below -50°C, below -20°C, below 0°C, or below 20°C for at least one week in the absence of moisture or other chemically reactive conditions.

[0048] The term "substituted," as used herein, means that a hydrogen radical of the specified moiety is replaced with the radical of a specified substituent, provided that the substitution results in a stable or chemically feasible compound. The term "substitutable," when used in reference to a specified atom, means that the bonded atom is a hydrogen radical and can be replaced with the radical of a suitable substituent.

[0049] The phrase "one or more substituents," as used herein, refers to a plurality of substituents ranging from one to the maximum number of possible substituents, based on the number of available bonding sites, provided that the stability and chemical feasibility conditions set forth above are met. Unless otherwise specified, an optionally substituted group may have a substituent at each substitutable position of the group, and the substituents may be the same or different.

[0050] As used herein, the terms "independently" or "independently selected" mean that the same or different values ​​may be selected for a given variable at multiple instances in a single compound.

[0051] An aryl (including the aryl portion of aralkyl, aralkoxy, aryloxyalkyl, etc.) or heteroaryl (including the heteroaryl portion of heteroaralkyl and heteroaralkoxy, etc.) group can contain one or more substituents. Examples of suitable substituents on the unsaturated carbon atom of an aryl or heteroaryl group include -halo, -NO, -CN, -R * , -C(R * )=C(R * )2, -C≡CR * , -OR * , -SR°, -S(O)R°, -SO2R°, -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°, -NR + CO2R°, -NR + SO2R°, -NR + SO2N(R + )2, -OC(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°)=N-OR * , -P(O)(R * )2, -P(O)(OR * )2, -OP(O)-OR * , and -P(O)(NR + )-N(R + )2, where R°, R + and R * is as defined above, or two adjacent substituents, together with intervening atoms, form a 5-6 membered unsaturated or partially unsaturated ring having 0-3 ring atoms selected from the group consisting of N, O, and S.

[0052] The aliphatic group or non-aromatic heterocyclic ring may be substituted with one or more substituents. Examples of suitable substituents on the saturated carbon of the aliphatic group or non-aromatic heterocyclic ring include those listed above for the unsaturated carbon of the aryl or heteroaryl group, as well as ═O, ═S, ═C(R * )2, =NN(R * )2, =N-OR * , =N-NHC(O)R * , =N-NHCO2Ro, = N-NHSO2Ro, or =NR * These include, but are not limited to, R * Oh Each of R and R° is as defined above.

[0053] Preferred substituents on a substitutable nitrogen atom of a heteroaryl or non-aromatic heterocycle are -R * , -N(R * )2, -C(O)R * , -CO2R * , -C(O)-C(O)R * -C(O)CHC(O)R * , -SO2R * , -SO2N(R * )2, -C(=S)N(R * )2, -C(=NH)-N(R * )2, and -NR * SO2R * Contains R * Each of the is as defined above. The ring nitrogen atom of a heteroaryl or non-aromatic heterocycle can also be oxidized to form the corresponding N-hydroxy or N-oxide compound. A non-limiting example of such a heteroaryl having an oxidized ring nitrogen atom is N-oxidopyridyl.

[0054] The term "about" is used herein to mean approximately, in the region of, roughly, or approximately. When the term "about" is used in conjunction with a numerical range, it modifies that range by extending the boundaries above and below the stated numerical values. In general, the term "about" is used herein to modify numerical values ​​above and below the stated value with a variance of 10%.

[0055] As used herein, the term "comprises" means "including, but not limited to."

[0056] It will be apparent to one of ordinary skill in the art that certain compounds of the present invention may exist in tautomeric forms, and all such tautomeric forms of the compounds are within the scope of the present invention. Unless otherwise stated, structures depicted herein are also meant to include all geometric (or conformational) isomers, i.e., (Z) and (E) double bond isomers and (Z) and (E) conformational isomers, in addition to all stereochemical forms of the structure, i.e., R and S configurations about each asymmetric center. Thus, enantiomeric and diastereomeric mixtures of the present compounds, as well as single stereochemical isomers, are within the scope of the invention. When the mixture is enriched in one stereoisomer over another, the mixture can contain, for example, an enantiomeric excess of at least 50%, 75%, 90%, 99%, or 99.5%.

[0057] Unless otherwise stated, structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the present structure include compounds having the replacement of a hydrogen atom with deuterium or tritium, or 13 C- or 14 Except for the replacement of a carbon atom with a C-rich carbon, it is within the scope of the present invention.

[0058] As used herein, the term "seeding" is used to refer to the addition of crystalline material to initiate crystallization or recrystallization.

[0059] When a compound crystallizes from a solution or slurry, it may crystallize in different spatial lattice arrangements, a phenomenon known as "polymorphism." Each crystalline form is a "polymorph." Polymorphs of a given substance have the same chemical composition, but they may differ from each other with respect to one or more physical properties, such as solubility and dissociation, true density, melting point, crystal shape, compaction behavior, flowability, and / or solid-state stability.

[0060] As used herein, the term "solvate" or "solvated" refers to a physical association of a compound with one or more solvent molecules. This physical association includes hydrogen bonding. In certain instances, a solvate is isolable, for example, when one or more solvent molecules are incorporated into the crystal lattice of a crystalline solid. "Solvate" encompasses both solution-phase and isolable solvates. Representative solvates include, for example, hydrates, ethanolates, or methanolates. The physical properties of solvates generally differ from other solvates and from unsolvated forms of the compound. Because the chemical composition also differs between solvates, these forms are referred to as "pseudopolymorphs."

[0061] As used herein, the term "hydrate" refers to a solvate in which the solvent molecule is present in a defined stoichiometric amount and may include, for example, a hemihydrate, monohydrate, dihydrate, or trihydrate. As used herein, the term "anhydrate" refers to a compound of the invention that does not contain HO incorporated into its crystal lattice.

[0062] As used herein, "crystal" refers to a solid having a highly general chemical structure. Specifically, a crystalline compound can be produced as one or more single crystalline forms of the compound. For purposes of this application, the terms "single crystalline form" or "crystalline form" can be used interchangeably to distinguish between crystals with different properties (e.g., different XRPD patterns, different DSC scan results). Thus, each distinct polymorph and pseudopolymorph of a compound is considered herein to be a distinct single crystalline form.

[0063] "Substantially crystalline" refers to a compound that can be at least a specified weight percent crystalline. The specified weight percent can be 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 percentage between 10% and 100%. In some 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.

[0064] "Substantially pure" refers to a compound that can be at least a specified weight percent of the compound. The specified weight percent can be 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%.

[0065] Unless otherwise expressly stated, structures depicted herein are meant to include all hydrates, anhydrates, solvates, and polymorphs thereof.

[0066] As used herein, the terms "Compound (I-1)" and "4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-methylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid" can be used interchangeably and include all crystalline forms. Both terms refer to the compound produced in Examples 1 and 1A of the following Examples, including both Form 1 and Form 2.

[0067] As used herein, the terms "Compound (I-1) Form 2" and "4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-methylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid Form 2" can be used interchangeably. Both terms refer to crystalline Form 2, as produced in Examples 1 and 1A of the Examples below.

[0068] As used herein, the terms "compound of formula (VIII-1)" and "(R)-1-((2,5-dichlorobenzamido)acetamido)-3-methylbutylboronic acid" can be used interchangeably. The compound of formula (VIII-1) is disclosed in U.S. Pat. No. 7,442,830 and WO 09 / 020448.

[0069] As used herein, the terms "compound of formula (I-15)," "compound (I-15)," and "(I-15)" can be used interchangeably and are used to refer to the citrate ester of compound (VIII-15), the compound produced in Example 15 of the Examples below.

[0070] As used herein, the term "anhydride," when used with respect to boronic acids, such as compounds of Formula (VIII), refers to a chemical 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 acid anhydride compound becomes hydrated, releasing the free boronic acid compound. In various embodiments, the boronic acid anhydride can contain two, three, four, or more boronic acid units and can have a cyclic or linear structure. Non-limiting examples of oligomeric boronic acids of peptide boronic acid compounds of the present invention are illustrated below.

[0071] [ka] In formulas (1) and (2), the variable nn is an integer from 0 to about 10, preferably 0, 1, 2, 3, or 4. In some embodiments, the boronic acid anhydride compound is a boronic acid anhydride having a ring structure of formula (2). The variable W has the formula (3):

[0072] [ka] In the formula, P, R a2 , A, R a1 , and R a is as defined herein.

[0073] As used herein, the total weight of a single oral pharmaceutical dosage form is determined by adding the weights of all of the ingredients in the oral pharmaceutical dosage form and does not include the weight of any coating that may optionally be applied to the oral pharmaceutical dosage form after it is formed. The total weight of the single oral pharmaceutical dosage form is used as the basis for calculating the weight percentage of each ingredient that comprises the oral pharmaceutical dosage form.

[0074] As used herein, "low moisture," when used with respect to excipients such as fillers, refers to excipients having a moisture content of about 0.5% to about 4%. The term "low moisture" may be used interchangeably with the term "low water."

[0075] As used herein, the terms "lyophilized powder," "cake," or "lyophilized cake" refer to any solid material obtained by freeze-drying an aqueous mixture.

[0076] As used herein, the term "tonicity modifier" refers to an agent that contributes to the osmolality of a fluid or solution.

[0077] As used herein, the terms "boronate ester" and "boronic ester" can be used interchangeably and refer to -B(Z1 )(Z 2 ) moiety, Z 1 and Z 2 together form a moiety in which the atom bonded to the boron is, in each case, an oxygen atom.

[0078] In some embodiments, the boronic ester moiety is a 5-membered ring. In some other embodiments, the boronic ester moiety is a 6-membered ring. In some other embodiments, the boronic ester moiety is a mixture of 5- and 6-membered rings.

[0079] As used herein, the term "α-hydroxycarboxylic acid" refers to a compound containing a hydroxy group directly bonded to a carbon atom alpha to the carboxylic acid group. As used herein, the term "α-hydroxycarboxylic acid" is not intended to be limited to compounds having only one hydroxy group and one carboxylic acid group.

[0080] As used herein, the term "β-hydroxycarboxylic acid" refers to a compound containing a hydroxy group attached directly to a carbon atom at the β-position relative to the carboxylic acid group. As used herein, the term "β-hydroxycarboxylic acid" is not intended to be limited to compounds having only one hydroxy group and one carboxylic acid group.

[0081] As used herein, the term "moiety derived from an α-hydroxycarboxylic acid" means It refers to a moiety formed by removing a hydrogen atom from a carboxylic acid in an α-hydroxycarboxylic acid and by removing a hydrogen atom from a hydroxy group that has a carbon atom directly bonded to the carboxylic acid group at the α-position. As used herein, the term "moiety derived from a β-hydroxycarboxylic acid" refers to a moiety formed by removing a hydrogen atom from a carboxylic acid in a β-hydroxycarboxylic acid and by removing a hydrogen atom from a hydroxy group that has a carbon atom directly bonded to the β-position. Detailed Description of the Invention In some embodiments, the alpha-hydroxy acid is characterized by formula (V):

[0082] [ka] In the formula, R b3 and R b4 is independently hydrogen, —COH, or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl group.

[0083] In some embodiments, R b3 and R b4 each independently represents hydrogen, C 1-6 Aliphatic, or -(CH2) p -COH, and p is 0, 1, or 2. In some embodiments, R b3 and R b4 each independently represents hydrogen or C 1-6 In certain such embodiments, R b3 and R b4 are independently selected from the group consisting of hydrogen, methyl, ethyl, isopropyl, isobutyl, tert-butyl, and cyclohexyl. b3 and R b4 each independently represents hydrogen or -(CH2) p In some such embodiments, p is 1. In certain other embodiments, R b3 and R b4 each independently represents -(CH2) p In certain such embodiments, p is 1.

[0084] In some embodiments, the α-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 benzilic acid. In some other embodiments, the α-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 benzilic acid. In particular embodiments, the α-hydroxycarboxylic acid is citric acid. Some other non-limiting examples of α-hydroxycarboxylic acids include glucoheptonic acid, gluconic acid (maltonic acid), lactobionic acid, and galactaric acid.

[0085] In some embodiments, the beta-hydroxy acid is characterized by formula (VI):

[0086] [ka] In the formula, R b1 and R b2 each independently represents hydrogen, -CO2H, -OH, or a substituent. is a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl group, and R b3 and R b4 each independently is hydrogen, —COH, or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl group; Or, R b2 and R b4 are each independently hydrogen, and R b1 and R b3together with the carbon atoms to which they are attached form an unsubstituted or substituted fused 4-8 membered non-aromatic ring having 0-3 ring heteroatoms selected from the group consisting of O, N, and S, which ring can be optionally fused to an unsubstituted or substituted 4-8 membered non-aromatic ring or a 5-6 membered aromatic ring having 0-3 ring heteroatoms selected from the group consisting of O, N, and S; or R b2 and R b4 does not exist, R b1 and R b3 together with the carbon atoms to which they are attached form an unsubstituted or substituted fused 5-6 membered aromatic ring having 0-3 ring heteroatoms selected from the group consisting of O, N, and S, which ring can be optionally fused to an unsubstituted or substituted 4-8 membered non-aromatic ring or a 5-6 membered aromatic ring having 0-3 ring heteroatoms selected from the group consisting of O, N, and S.

[0087] In some embodiments, R b1 and R b2 each independently represents hydrogen, C 1-6 Aliphatic, -(CH2) p -OH, or -(CH2) p -COH, and p is 0, 1, or 2. In some such embodiments, R b1 and R b2 is hydrogen. In some other such embodiments, R b1 is -OH and R b2 is hydrogen.

[0088] In some embodiments, R b3 and R b4 each independently represents hydrogen, C 1-6 Aliphatic, or -(CH2) p -COH, and p is 0, 1, or 2. In some embodiments, R b3 and R b4 each independently represents hydrogen or C 1-6 In certain such embodiments, Rb3 and R b4 is independently selected from the group consisting of hydrogen, methyl, ethyl, isopropyl, isobutyl, tert-butyl, and cyclohexyl. b3 and R b4 each independently represents -(CH2) p -CO2H, and p is 0 or 1.

[0089] The variable p is 0, 1, or 2. In some embodiments, p is 0 or 1. In particular embodiments, p is 0. In other embodiments, p is 1.

[0090] In some embodiments, R b2 and R b4 does not exist, R b1 and R b3 together with the carbon atoms to which they are attached form a substituted or unsubstituted phenyl ring.

[0091] In some 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 some 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 particular embodiments, the β-hydroxycarboxylic acid is citric acid. Some other non-limiting examples of β-hydroxycarboxylic acids include glucoheptonic acid, gluconic acid (maltonic acid), lactobionic acid, and galactaric acid. Some other non-limiting examples of β-hydroxycarboxylic acids include embonic acid, 1-hydroxy-2-naphthoic acid, and 3-hydroxy-2-naphthoic acid.

[0092] In some embodiments, the alpha-hydroxy acid or beta-hydroxy acid is glycolic acid, malic acid, hexahydromandelic acid, 2-hydroxyisobutyric acid, citric acid, mandelic acid, lactic acid, 3-hydroxybutyric acid, beta-hydroxyisovaleric acid, 2-hydroxy-3,3-dimethylaminopropyl methyl ester. The hydroxybenzoic acid is selected from the group consisting of 2-hydroxy-3-methylbutyric acid, 2-hydroxyisocaproic acid, tartaric acid, salicylic acid, and benzilic acid.

[0093] In some embodiments, the compound of general formula (I) is characterized by formula (II):

[0094] [ka] During the ceremony, 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 has the value described above.

[0095] In some embodiments, R b1 , R b2 , R b3 , and R b4 Any one of may contain a functional group capable of forming an additional bond with the boron atom. In certain embodiments, the functional group is a carboxylic acid. In other certain embodiments, the functional group is a hydroxyl group.

[0096] In some embodiments, the α-hydroxycarboxylic acid or β-hydroxycarboxylic acid is citric acid, and the compound of general formula (I) is represented by formula (III) or (IV):

[0097] [ka] or a mixture thereof, characterized by the variables P, A, R a , R a1 , and Ra2 has the values ​​described below.

[0098] In some other embodiments, the α-hydroxycarboxylic acid or β-hydroxycarboxylic acid is citric acid, and an additional bond can be formed between the carboxylic acid of formula (III) or (IV) and the boron atom. Without being bound by any chemical bonding theory, in such embodiments, the compound of general formula (I) can be represented by formula (IIIa) or (IVa):

[0099] [ka] or a mixture thereof, with the variables P, A, R a , R a1 , and R a2 has the values ​​described below.

[0100] Without being bound by any theory of chemical bonding, it is recognized that there are other representations that can be used to indicate this additional bond between the boron atom and the carboxylic acid in formulas (IIIa) and (IVa).

[0101] The following values ​​appear as variables in any of formulas (I), (II), (III), (IIIa), (IV), or (IVa):

[0102] The variable P is hydrogen or an amino-blocking moiety. Non-limiting examples of amino-blocking moieties are described in P.G.W. Buts and T.W. Greene, "Greene's Protective Groups in Organic Synthesis" (4 th ed.), John Wiley & Sons, NJ (2007), and can include, for example, acyl, sulfonyl, oxyacyl, and aminoacyl groups.

[0103] In some embodiments, 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-, and R c is C 1-6 aliphatic, C 1-6 Fluoroaliphatic, -R D , -T 1 -R D , and -T 1 -R 2c and the variable T 1 , R D , R 2c , and R 4c has the values ​​described below.

[0104] Variable R 4c is hydrogen, C 1-4 Alkyl, C 1-4 Fluoroalkyl, or C 6-10 Ara (C 1-4 alkyl), wherein the aryl portion is substituted or unsubstituted. In some embodiments, R 4c is hydrogen or C 1-4 In certain embodiments, R 4c is hydrogen.

[0105] Variable T 1 are 0 to 2 independently selected R 3a or R 3b C replaced by 1-6 is an alkylene chain, and the alkylene chain is optionally —C(R 5 )=C(R 5 )-, -C≡C-, or -O-. 3a are independently -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). 3b are independent, R 3a or R 7 C replaced by1-3 aliphatic or two substituents R on the same carbon atom 3b together with the carbon atoms to which they are attached form a 3- to 6-membered cycloaliphatic ring. 7 is a substituted or unsubstituted aromatic group. In some embodiments, T 1 is C 1-4 Archi It is a Ren chain.

[0106] Variable R 2c , 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(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, Each R 4 are independently hydrogen, or an optionally substituted aliphatic, aryl, heteroaryl, or heterocyclyl group, or two R on the same nitrogen atom 4 together with the nitrogen atom form a substituted or unsubstituted 4- to 8-membered heterocyclyl ring which, in addition to the nitrogen atom, has 0 to 2 ring heteroatoms independently selected from the group consisting of N, O, and S; Each R 5 are independently hydrogen or an optionally substituted aliphatic, aryl, heteroaryl, or heterocyclyl group; Each R 6is independently an optionally substituted aliphatic, aryl, or heteroaryl group.

[0107] Variable R D is a substituted or unsubstituted aromatic, heterocyclyl, or cycloaliphatic ring, any of which is optionally fused to a substituted or unsubstituted aromatic, heterocyclyl, or cycloaliphatic ring. In some embodiments, R D is 0 to 2 R d and 0 to 2 R 8d substituted on a substitutable ring carbon atom with R D Each substitutable ring nitrogen atom of is unsubstituted or is —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 Al (C 1-4 ) alkyl, and the aryl portion of these may be substituted or unsubstituted. 4 , R 5 , and R 6 has the value described above. d independently, C 1-6 aliphatic, C 1-6 Fluoroaliphatic, Halo, -R 1d , -R 2d , -T 2 -R 1d , and -T 2 -R 2d and the variable T 2 , R 1d , R 2d , and R 8d has the value described below. In some embodiments, each R d independently, C 1-6 aliphatic, C 1-6 is selected from the group consisting of fluoroaliphatic, and halo.

[0108] T 2 is 0 to 2 independently selected R3a or R 3b C replaced by 1-6 an alkylene chain, wherein the alkylene chain is optionally —C(R 5 )=C(R 5 )-, -C≡C-, or -O-. Variable R 3a , R 3b , and R 5 has the value described above.

[0109] Each R 1d is independently a substituted or unsubstituted aryl, heteroaryl, heterocyclyl, or cycloaliphatic ring.

[0110] Each R 2d are independently -NO2, -CN, -C(R 5 )=C(R 5 )2, -C≡CR 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, -OC(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. Variable R 4 , R 5 , and R 6 has the values ​​described below.

[0111] Each R 8d independently, 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 In some embodiments, each R 8d independently, C 1-4 aliphatic, C 1-4 fluoroaliphatic, or halo.

[0112] In some embodiments, R D is a substituted or unsubstituted monocyclic or bicyclic ring system. In some embodiments, R D is a substituted or unsubstituted monocyclic or bicyclic ring system selected from the group consisting of furanyl, thienyl, pyrrolyl, isoxazolyl, oxazolyl, thiazolyl, isothiazolyl, imidazolyl, pyrazolyl, oxadiazolyl, thiadiazolyl, phenyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, benzofuranyl, benzothiophenyl, indolyl, benzoxazolyl, benzisoxazolyl, benzimidazolyl, indazolyl, purinyl, naphthyl, quinolinyl, isoquinolinyl, cinnolinyl, quinazolinyl, quinoxalinyl, phthalazinyl, naphthyridinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, tetrahydroquinoxalinyl, and dihydrobenzoxazinyl. Dis a substituted or unsubstituted monocyclic or bicyclic ring system selected from the group consisting of phenyl, pyridinyl, pyrimidinyl, pyrazinyl, naphthyl, benzimidazolyl, quinolinyl, isoquinolinyl, quinoxalinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, tetrahydroquinoxalinyl, and dihydrobenzoxazinyl.

[0113] In some embodiments, R D The substitutable ring carbon atoms of d and 0 to 2 R 8d substituted on a substitutable carbon atom with Each R d independently, C 1-6 aliphatic, C 1-6 fluoroaliphatic, or halo; Each R 8d independently, C 1-4 aliphatic, C 1-4 fluoroaliphatic, or halo.

[0114] In some embodiments, R D The substitutable ring carbon atoms of d and 0 to 2 R 8d and wherein T 1 is unsubstituted or R 3a or R 3b Replaced by C 1-3 is an alkylene chain, Each R 1d is independently a substituted or unsubstituted aryl, heteroaryl, heterocyclyl, or cycloaliphatic ring; Each R 2d are independently -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, -OC(O)R 5 , -OC(O)N(R 4 )2, -C(O)R 5 , -CO2R 5 , or -C(O)N(R 4 )2. Variable R 4 , R 5 , and R 6 has the value described above.

[0115] In some embodiments, the variable R d is the formula -QR E Q is -O-, -NH-, or -CH2-; R E is a substituted or unsubstituted aryl, heteroaryl, heterocyclyl, or cycloaliphatic ring. E is a substituted or unsubstituted phenyl, pyridinyl, pyrimidinyl, pyrazinyl, piperidinyl, piperazinyl, or morpholinyl ring.

[0116] In some embodiments, P has the formula R c -C(O)-, and R c is C 1-4 Alkyl, C 1-4 Fluoroalkyl, or C 6-10 Ara (C 1-4 ) alkyl, these The aryl moiety is substituted or unsubstituted. In certain such embodiments, P is selected from the group consisting of acetyl, trifluoroacetyl, and phenylacetyl.

[0117] In some other embodiments, P has the formula R D -C(O)-, and R D is a substituted or unsubstituted phenyl, pyridinyl, pyrazinyl, pyrimidinyl, quinolinyl, or quinoxalinyl. In yet some other embodiments, P is a group of formula R D -C(O) and R D is 0 to 1 R d and 0 to 2 R 8d phenyl, pyridinyl, pyra, The aryl group is preferably benzoxazinyl, pyrimidinyl, naphthyl, quinolinyl, quinoxalinyl, benzimidazolyl, or dihydrobenzoxazinyl.

[0118] In certain embodiments, P has the formula R D -C(O)-, and R D is 2-pyrazinyl. In other particular embodiments, P is a group of formula R D -C(O)-, and R D is 2,5-dichlorophenyl. In yet other particular embodiments, P is a group of formula R D -C(O)-, and R D is 6-phenyl-2-pyridinyl.

[0119] In some other embodiments, P has the formula R c -SO2-, R c -R D or -T 1 -R D and T 1 is C 1-4 alkylene, and R D is 0 to 1 R d and 0 to 2 R 8d and n is 0 or 1. The group is phenyl, pyridinyl, pyrazinyl, pyrimidinyl, naphthyl, quinolinyl, quinoxalinyl, benzimidazolyl, or dihydrobenzoxazinyl, substituted by

[0120] 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 and the variable R 4 , R 5 , and R 6 has the value given above, and the variable R 5a , R 5b , R B , Y, and m have the values ​​described below.

[0121] In some embodiments, R a is hydrogen, C 1-6 aliphatic, C 1-6 Fluoroaliphatic, or -(CH2) m -CH2-R B In some other embodiments, R a is C 1-6 Aliphatic, or -(CH2) m -CH2-R B In some further embodiments, R a is C 1-6 In still other further embodiments, R a is isobutyl, 1-naphthylmethyl, 2-naphthylmethyl, benzyl, 4-fluorobenzyl, 4-hydroxybenzyl, 4-(benzyloxy)benzyl, benzylnaphthylmethyl, or phenethyl. a is isobutyl.

[0122] 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 and the variable R 4 , R 5 , and R 6 has the value given above, and the variable R 5a , R 5b , R B , Y, and m have the values ​​described below.

[0123] In some 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 some other embodiments, R a1 is hydrogen, -(CH2) m -CH2-R B , or -(CH2) m -CH(R 5a )-OR 5b In some other embodiments, R a1 is isobutyl, 1-naphthylmethyl, 2-naphthylmethyl, benzyl, 4-fluorobenzyl, 4-hydroxybenzyl, 4-(benzyloxy)benzyl, benzylnaphthylmethyl, or phenethyl.

[0124] In certain embodiments, R a1 is -CH2-R B In other particular embodiments, R a1 is -CH(R 5a )-OR 5b In still other particular embodiments, R a1 is hydrogen.

[0125] Variable R a2 is hydrogen, C1-6 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 and the variable R 4 , R 5 , and R 6 has the value given above, and the variable R B , R 5a , R 5b , Y, and m have the values ​​described below.

[0126] In some embodiments, R a2 is hydrogen, C 1-6 aliphatic, C 1-6 Fluoroaliphatic, -(CH2) m -CH2-R B , or -(CH2) m -CH(R 5a )-OR 5b In some other embodiments, R a2 is isobutyl, 1-naphthylmethyl, 2-naphthylmethyl, benzyl, 4-fluorobenzyl, 4-hydroxybenzyl, 4-(benzyloxy)benzyl, benzylnaphthylmethyl, or phenethyl.

[0127] Each R B is independently a substituted or unsubstituted monocyclic or bicyclic ring system. In some embodiments, each R Bis independently a substituted or unsubstituted phenyl, pyridyl, indolyl, benzimidazolyl, naphthyl, quinolinyl, quinoxalinyl, or isoquinolinyl ring. B is a substituted or unsubstituted phenyl ring.

[0128] The variable Y is hydrogen, —CN, or —NO 2 . In some embodiments, Y is —NO 2 .

[0129] Variable R 5a is hydrogen, or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl group. In some embodiments, R 5a is hydrogen or a substituted or unsubstituted aliphatic group. In some other embodiments, R 5a is hydrogen, or C 1-6 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.

[0130] Variable R 5b is hydrogen, or a substituted or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl group. In some embodiments, R 5b is hydrogen or a substituted or unsubstituted aliphatic group. In some other embodiments, R 5b is hydrogen or C 1-6 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.

[0131] The variable m is 0, 1, or 2. In some embodiments, m is 0 or 1. In certain embodiments, m is 0. In other certain embodiments, m is 1.

[0132] The variable A is 0, 1, or 2. In some embodiments, A is 0 or 1. In certain embodiments, A is 0.

[0133] The variable n is 0 or 1. In certain embodiments, n is 0. In other certain embodiments, n is 1.

[0134] In some embodiments, A is 0 and R a is hydrogen, C 1-6 aliphatic, C 1-6 Fluoroaliphatic, or -(CH2) m -CH2-R B and 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 and P is R c -C(O)- or R c -S(O)2- and R c -R D and m is 0 or 1.

[0135] In some other embodiments, A is 0 and R a is C 1-6 Aliphatic or -(CH2) m -CH2-R B and R a1 is hydrogen, -(CH2) m -CH2-R B ,or -(CH2) m -CH(R 5a )-OR 5b and P is R c -C(O)- or R c -S(O)2- and R c -R D and m is 0 or 1.

[0136] In some other embodiments, A is 0 and Ra is C 1-6 It is aliphatic and R a1 is hydrogen, -(CH2) m -CH2-R B , or -(CH2) m -CH(R 5a )-OR 5b and P is R c -C(O) and R c -R D and m is 0 or 1.

[0137] In some other embodiments, A is 0 and R a is isobutyl, and 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 and P is R c -C(O)- and R c -R D and m is 0 or 1.

[0138] In still some other embodiments, A is 0 and R a is isobutyl, and R a1 is hydrogen, C 1-6 Aliphatic, -(CH2) m -CH2-R B , or -(CH2) m -CH(R 5a )-OR 5b and P is R c -C(O)- and R c -R D and m is 0 or 1.

[0139] In still yet some other embodiments, A is 0 and R a is isobutyl, and R a1 is hydrogen, -(CH2) m -CH2-R B , or -(CH2)m -CH(R 5a )-OR 5b and P is R c -C(O)- and R c -R D and m is 0 or 1.

[0140] In certain embodiments, A is 0 and R a is isobutyl, and R a1 is -CH2-R B and R B is phenyl and P is R c -C(O)- and R c -R D and R D is 2-pyrazinyl.

[0141] In other particular embodiments, A is 0 and R a is isobutyl, and R a1 is hydrogen and P is R c -C(O)- and R c -R D and R D is 2,5-dichlorophenyl.

[0142] In yet other particular embodiments, A is 0 and R a is isobutyl, and R a1 is -CH(R 5a )-OR 5b and R 5a is C 1-6 It is aliphatic and R 5b is hydrogen and P is R c -C(O)- and R c -R D and R D is 6-phenyl-2-pyridinyl-.

[0143] In some embodiments, the compound of formula (I) has the formula (I-1):

[0144] [ka] or characterized by its crystalline form.

[0145] In some other embodiments, the compound of Formula (I) has the formula (I-15):

[0146] [ka] or characterized by its crystalline form.

[0147] In still some other embodiments, the compound of formula (I) has formula (I-18):

[0148] [ka] or characterized by its crystalline form.

[0149] General synthetic methodology The compound of formula (I) can be prepared by esterification of corresponding boronic acid. Such boronic acid compounds can be prepared by methods known to those skilled in the art. For example, see U.S. Patent No. 5,780,454 to Adams et al. and International Publication No. WO 2005 / 097809 to Pickersgill et al. An exemplary synthetic route is described in Scheme 1 below.

[0150] Scheme 1:

[0151] [ka] Coupling of compound i with an N-protected amino acid ii, followed by N-terminal deprotection, provides compound iii or a salt thereof. Examples of suitable protecting groups (PG) include acyl protecting groups such as formyl, acetyl (Ac), succinyl (Suc), and methoxysuccinyl. Protecting groups include, but are not limited to, urethane protecting groups such as, for example, tert-butoxycarbonyl (Boc), benzyloxycarbonyl (Cbz), and fluorenylmethoxycarbonyl (Fmoc). Optionally, PG is hydrogen, and deprotection is not required. The peptide coupling reaction can be carried out by prior conversion of the carboxylic acid moiety of compound ii to an activated ester or acid halide, such as, for example, an O-(N-hydroxysuccinimide) ester, followed by treatment with compound i. Alternatively, the activated ester can be generated in situ by contacting the carboxylic acid with a peptide coupling reagent. Examples of suitable peptide coupling reagents include, but are not limited to, carbodiimides such as, for example, dicyclohexylcarbodiimide (DCC) or 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC), phosphonium reagents such as, for example, (benzotriazol-1-yloxy)tris(dimethylamino)phosphonium hexafluorophosphate (BOP), and uronium reagents such as, for example, O-(1H-benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium tetrafluoroborate (TBTU).

[0152] Compound iii is then coupled with an amino-blocking moiety to give compound iv. The peptide coupling conditions described above for coupling compounds i and ii are also suitable for coupling compound iii with an amino-blocking moiety. Deprotection of the boronic acid moiety then gives compound v. The deprotection step preferably involves the coupling of a boronic ester compound iv, an organoboronic acid acceptor, a lower alkanol, C 5-8 This is accomplished by transesterification in a biphasic mixture containing a hydrocarbon solvent and an aqueous mineral acid. Other reagents that can be used to deprotect the boronic acid moiety include, but are not limited to, BCl3, lithium aluminum hydride, and NaIO4.

[0153] Scheme 2:

[0154] [ka] Alternatively, the order of coupling reactions can be reversed, as shown in Scheme 2. Thus, O-protected amino acid vi is first coupled with an amino-blocking moiety, followed by ester hydrolysis to form compound vii. Optionally, PG' is H, which does not require ester hydrolysis and directly leads to compound vii. Coupling with compound i and boronic acid deprotection are accomplished as described above for Scheme 1 to give compound v.

[0155] As shown in Scheme 3, compound v is reacted with an appropriate α-hydroxycarboxylic acid or β-hydroxycarboxylic acid to give compounds of formula (I).

[0156] Scheme 3:

[0157] [ka] Conversion of v to a compound of Formula (I) can be accomplished under esterification conditions employing approximately a molar equivalent of an α-hydroxycarboxylic acid or β-hydroxycarboxylic acid to a solvent such as ethyl acetate at a temperature of about 40°C to about 80°C. Conversion of v to a compound of Formula (I) can also employ a molar excess of the α-hydroxycarboxylic acid or β-hydroxycarboxylic 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, methylcyclohexane, tert-butyl methyl ether, and mixtures thereof. The choice of solvent may depend, in part, on the solubility of the α-hydroxycarboxylic acid or β-hydroxycarboxylic acid used. The temperature selected for the conversion of v to a compound of Formula (I) may depend, in part, on the boiling point of the solvent or solvent mixture used.

[0158] The conversion of 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-tertbutylpyridine, 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.

[0159] The compound of formula (v) and an α-hydroxycarboxylic acid or β-hydroxycarboxylic acid are heated together in a suitable solvent for a period of time. After this period, the reaction mixture is allowed to cool for a period of time, and the compound of formula (I), which precipitates upon cooling, is collected by filtration. The cooling can be uncontrolled or can be controlled by using 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 and then evaporating the solvent. The reaction mixture can be seeded with crystals of the compound of formula (I) to cause precipitation.

[0160] A co-solvent, such as, but not limited to, heptane, methylcyclohexane, toluene, tert-butyl methyl ether, ethyl acetate, or a mixture thereof, can be added during the cooling period. After the addition of the co-solvent, the reaction mixture can be cooled, further resulting in precipitation of the compound of Formula (I). Alternatively, upon addition of the co-solvent, the reaction mixture can then be heated again to produce a homogeneous solution, which is then cooled, resulting in precipitation of the compound of Formula (I). The reaction mixture can be seeded with crystals of the compound of Formula (I) to produce the precipitation.

[0161] 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% or more. , about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or about 99.5%.

[0162] In some embodiments, the compound of Formula (I) is isolated in crystalline form. In some embodiments, the compound of Formula (I) is isolated in substantially crystalline form. In some other embodiments, the compound of Formula (I) is isolated in amorphous form.

[0163] The compound of formula (I) can also be produced by co-lyophilizing compound v and an α-hydroxycarboxylic acid or a β-hydroxycarboxylic acid. This is achieved by subjecting an aqueous solution containing the compound of formula v and a molar excess of an α-hydroxycarboxylic acid or a β-hydroxycarboxylic acid to a lyophilization procedure. In some embodiments, the aqueous solution additionally contains a water-miscible cosolvent. Examples of suitable cosolvents include, but are not limited to, tert-butyl alcohol, methanol, ethanol, and mixtures thereof. Co-lyophilization produces a composition containing the compound of formula (I) and an excess of an α-hydroxycarboxylic acid or a β-hydroxycarboxylic acid.

[0164] Use, Formulation, and Administration The present invention provides compounds that are potent inhibitors of the proteasome. Compounds can be assayed in vitro or in vivo for their ability to inhibit proteasome-mediated peptide hydrolysis or protein degradation.

[0165] In another aspect, therefore, the present invention provides a method for inhibiting one or more peptidase activities of the proteasome in a cell, comprising contacting a cell in which proteasome inhibition is desired with a compound described herein, or a pharmaceutically acceptable salt, boronic ester, or boronic anhydride thereof.

[0166] The present invention also provides a method for inhibiting cell proliferation, which includes contacting a compound described herein with cells in which such inhibition is desired. The phrase "inhibiting cell proliferation" is used to indicate the ability of a compound of the present invention to inhibit cell number or cell proliferation in contacted cells compared to cells not contacted with the inhibitor. Cell proliferation can be assessed using a cell counter or by a cell viability assay, such as an MTT or WST assay. When cells are undergoing solid growth (e.g., solid tumors or organs), such assessment of cell proliferation can be achieved by measuring proliferation using, for example, a caliper and comparing the magnitude of proliferation of contacted cells with that of non-contacted cells.

[0167] Preferably, the proliferation of cells contacted with the inhibitor is slowed by at least about 50% compared to the proliferation of non-contacted cells. In various embodiments, the proliferation of contacted cells is inhibited by at least about 75%, at least about 90%, or at least about 95% compared to non-contacted cells. In some embodiments, the phrase "inhibiting cell proliferation" includes a reduction in the number of contacted cells compared to non-contacted cells. Thus, proteasome inhibitors that inhibit cell proliferation in contacted cells can induce the contacted cells to undergo growth retardation, growth arrest, programmed cell death (i.e., apoptosis), or necrotic cell death.

[0168] In another aspect, the present invention provides a pharmaceutical composition comprising a compound of formula (I), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0169] In some embodiments, the composition also includes a free α-hydroxycarboxylic acid or salt thereof, or a β-hydroxycarboxylic acid or salt thereof. In such embodiments, the α-hydroxycarboxylic acid or salt thereof, or the β-hydroxycarboxylic acid or salt thereof The salt and the compound of Formula (I) are present in a molar ratio ranging from about 2:1 to about 200:1. In various embodiments, the α-hydroxycarboxylic acid or salt thereof, or β-hydroxycarboxylic acid or salt thereof, and the compound of Formula (I) are present in a ratio ranging from about 2:1 to about 200:1, from about 15:1 to about 80:1, or from about 20:1 to about 40:1.

[0170] When pharmaceutically acceptable salts of the compounds of the present invention are utilized in these compositions, the salts are preferably derived from inorganic or organic acids or bases. For reviews of suitable salts, see, for example, Berge et al., J. Pharm. Sci. 66:1-19 (1977), and Remington: The Science and Practice of Pharmacy, 20th Ed., ed. A. Gennaro, Lippincott Williams & Wilkins, 2000.

[0171] Non-limiting examples of suitable acid addition salts include acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, fumarate, lucoheptanoate, glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, oxalate, pamoate, pectinate, persulfate, 3-phenyl-propionate, picrate, pivalate, propionate, succinate, tartrate, thiocyanate, tosylate, undecanoate, and the like.

[0172] 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 such as zinc salts, salts with organic bases (e.g., dicyclohexylamine salts, N-methyl-D-glucamine, t-butylamine, ethylenediamine, ethanolamine, and choline), and salts with amino acids (e.g., arginine, lysine).

[0173] The term "pharmaceutically acceptable carrier" is used herein to refer to a substance that is compatible with a recipient subject, preferably a mammal, more preferably a human, and is suitable for delivering an active agent to a target site without terminating the activity of the agent. Any toxicity or side effects associated with the carrier are preferably commensurate with a reasonable risk / benefit ratio for the intended use of the active agent.

[0174] The terms "carrier," "excipient," or "vehicle" are used interchangeably herein and include any and all of solvents, diluents, and other liquid media, dispersing or suspending aids, surfactants, pH adjusters, isotonicity agents, thickeners or emulsifiers, preservatives, solid binders, lubricants, etc., as appropriate for the specific dosage form desired. Remington: The Science and Practice of Pharmacy, 20th Ed., ed. A. Gennaro, Lippincott Williams & Wilkins, 2000, discloses various carriers used to formulate pharmaceutically acceptable compositions, as well as known techniques for their preparation. Strickley, Pharmaceutical Research, 21(2)201-230(2004), reviews pharmaceutically acceptable excipients used in commercial products to solubilize compounds for oral or parenteral administration. Except insofar as any conventional carrier medium is incompatible with the compounds of the present invention, such as by producing any undesired biological effects or otherwise interacting in a deleterious manner with any other component of the pharmaceutically acceptable composition, its use is contemplated within the scope of the present invention. Some examples of substances that can act as buffers include ion exchangers, alumina, aluminum stearate, lecithin, serum proteins (e.g., human serum albumin), buffer substances (e.g., phosphates, carbonates, magnesium hydroxide and aluminum hydroxide, glycine, sorbic acid, or potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, pyrogen-free water, and the like. water), salts or electrolytes (e.g., protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, and zinc salts), colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, polyacrylates, waxes, polyethylene-polyoxypropylene block polymers, wool fat, sugars (such as lactose, glucose, sucrose, and mannitol), starches (e.g., corn starch and potato starch), cellulose and its derivatives (e.g., sodium carboxymethylcellulose, ethyl cellulose, and cellulose acetate), powdered tragacanth gum, malt, gelatin, talc, excipients (e.g., cocoa butter and suppository wax), oils (e.g., peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil), gluten, Examples of suitable anti-inflammatory agents include, but are not limited to, glycols (e.g., propylene glycol and polyethylene glycol), esters (e.g., ethyl oleate and ethyl laurate), agar, alginic acid, isotonic saline, Ringer's solution, alcohols (e.g., ethanol, isopropyl alcohol, ethyl alcohol, hexadecyl alcohol, and glycerol), cyclodextrins (e.g., hydroxypropyl β-cyclodextrin and sulfobutyl ether β-cyclodextrin), lubricants (e.g., sodium lauryl sulfate and magnesium stearate), and petroleum hydrocarbons (e.g., mineral oil and petrolatum). Coloring agents, release agents, coating agents, sweetening agents, flavoring agents, and fragrances, preservatives, and antioxidants can also be present in the composition, according to the judgment of the formulator.

[0175] The pharmaceutical compositions of the present invention can be manufactured by methods known in the art, such as conventional granulating, mixing, dissolving, encapsulating, lyophilizing, or emulsifying processes. The compositions can be produced in a variety of dosage forms, including granules, precipitates, or granules, powders (including lyophilized, rotary dried, or spray dried powders), amorphous powders, tablets, capsules, syrups, suppositories, injections, emulsions, elixirs, suspensions, or solutions.

[0176] According to a preferred embodiment, the compositions of the present invention are formulated for pharmaceutical administration to mammals, preferably humans. Such pharmaceutical compositions of the present invention can be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, bucally, vaginally, or via an implanted reservoir. As used herein, the term "parenteral" includes injection or infusion techniques such as subcutaneous, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrathecal, intrahepatic, intralesional, and intracranial. Preferably, the compositions are administered orally, parenterally, or subcutaneously. The formulations of the present invention can be designed to be short-acting, fast-releasing, or long-acting. Furthermore, the compounds can be administered locally rather than systemically, for example, at a tumor site (e.g., by injection).

[0177] Liquid dosage forms for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs.In addition to active compounds, liquid dosage forms can contain inert diluents commonly used in the art, such as water or other solvents, solubilizers and emulsifiers (e.g., ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, cyclodextrin, dimethylformamide), oils (especially cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol and sorbitan fatty acid esters, and their mixtures.In addition to inert diluents, oral compositions can also contain wetting agents, emulsifiers and suspensions. Adjuvants, such as flavoring, sweetening, and perfuming agents, may also be included.

[0178] Injectable preparations, for example, sterile injectable aqueous or oily suspensions, can be formulated by conventional methods using suitable dispersing or wetting agents and suspending agents. Sterile injectable preparations can also be sterile injectable solutions, suspensions, or emulsions in parenterally acceptable diluents or solvents, for example, as solutions in 1,3-butanediol. Acceptable vehicles and solvents that can be used include water, Ringer's solution, USP, and isotonic saline. In addition, sterile, fixed oils are conventionally used as solvents or suspending media. For this purpose, any bland fixed oil, including synthetic mono- or diglycerides, can be used. In addition, fatty acids such as oleic acid and its glyceride derivatives are used in the preparation of injectables. Injectable preparations can be sterilized by incorporating a sterilizing agent in the form of a sterile solid composition that can be dissolved or dispersed in sterile water or other sterile injectable media before use, for example, by filtration through a bacterial-retaining filter. Compositions formulated for parenteral administration may be injected by bolus or timed push, or may be administered by continuous infusion.

[0179] Oral solid dosage forms include capsules, tablets, pills, powders, and granules.In these solid dosage forms, the active compound is mixed with at least one inert pharmaceutically acceptable excipient or carrier, such as sodium citrate or dibasic calcium phosphate, and / or a) filler or extender, such as starch, lactose, cellulose, sucrose, glucose, mannitol, and silicic acid; b) binder, such as carboxymethylcellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, and acacia; c) humectant, such as glycerol; d) agar-agar, calcium carbonate, potato or tapioca starch, alginate, certain silicates, crospolis, The pharmaceutical composition may be mixed with disintegrating agents such as vidone, cellulose, croscarmellose sodium, 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 fumarate, stearic acid, solid polyethylene glycol, sodium lauryl sulfate, glycerin borate, and mixtures thereof. In the case of capsules, tablets, and pills, the dosage forms may also contain buffering agents such as phosphates or carbonates.

[0180] Solid compositions of a similar type can also be used as fillers in soft and hard gelatin capsules using excipients such as lactose or milk sugar and high molecular weight polyethylene glycols. Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells, such as enteric coatings and other coatings known in the pharmaceutical arts. 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 used as fillers in soft and hard gelatin capsules using excipients such as lactose or milk sugar and high molecular weight polyethylene glycols.

[0181] Active compound can also be in the form of microencapsulation, with one or more excipients as mentioned above.Solid dosage forms such as tablets, dragees, capsules, pills and granules can be prepared with coatings and shells, such as enteric coatings, release-controlling coatings and other coatings known in the pharmaceutical industry.Such solid dosage forms can be prepared with coatings and shells, such as enteric coatings, release-controlling coatings and other coatings known in the pharmaceutical industry. In dosage forms, the active compound can be mixed with at least one inert diluent, such as sucrose, lactose, or starch. Such dosage forms can also contain additional substances other than inert diluents, such as tableting lubricants and other tableting aids, such as magnesium stearate and microcrystalline cellulose, as is common practice. In the case of capsules, tablets, and pills, dosage forms can also contain buffering agents. They can optionally contain opacifying agents and can be of a composition that releases 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 some embodiments, excipients or carriers include, but are not limited to, sodium stearate fumarate, carboxymethylcellulose, magnesium stearate, crospovidone, ethylcellulose, talc, and silicified microcrystalline cellulose.

[0182] Dosage forms for topical or transdermal administration of the compounds of the present invention include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants, or patches. The active ingredient is mixed under sterile conditions with a pharmaceutically acceptable carrier and any necessary preservatives or buffers that may be required. Ophthalmic formulations, ear drops, and eye drops are also contemplated within the scope of the present invention. Additionally, the present invention contemplates the use of transdermal patches, which have the added advantage of providing controlled delivery of the compound to the body. Such dosage forms can be prepared by dissolving or dispersing the compound in a suitable medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate can be controlled by providing a rate-controlling membrane or by dispersing the compound in a polymer matrix or gel.

[0183] In some embodiments, the present invention provides pharmaceutical compositions comprising a compound of Formula (I) and additional excipients described herein. In some other embodiments, the present invention provides pharmaceutical compositions comprising a compound of Formula (II) and additional excipients described herein. In yet some other embodiments, the present invention provides pharmaceutical compositions comprising a compound of Formula (III) or (IV) and additional excipients described herein.

[0184] In a further embodiment, the present invention provides a pharmaceutical composition comprising the citrate ester of compound (VIII-1) and an additional excipient as described herein. In another further embodiment, the present invention provides a pharmaceutical composition comprising the citrate ester of compound (VIII-15) and an additional excipient as described herein. In a further embodiment, the present invention provides a pharmaceutical composition comprising the citrate ester of (VIII-18) and an additional excipient as described herein.

[0185] [ka] In further embodiments, the present invention provides pharmaceutical compositions comprising compound (I-1), or a crystalline form thereof. In still further embodiments, the present invention provides pharmaceutical compositions comprising compound (I-15), or a crystalline form thereof. In still yet some further embodiments, the present invention provides pharmaceutical compositions comprising compound (I-18), or a crystalline form thereof.

[0186] The following description of pharmaceutical compositions and methods for preparing said pharmaceutical compositions is applicable to compounds of formula (I), (II), (III), (IIIa), (IV), or (IVa) and the various embodiments of these formulas described herein. The following description of pharmaceutical compositions and methods for preparing said pharmaceutical compositions is also applicable to compounds (I-1), (I-15), or (I-18).

[0187] 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 yet another embodiment, the compound of formula (I) in the pharmaceutical composition is in crystalline form.

[0188] In some embodiments, the pharmaceutical formulations of the present invention provide stable, solid oral dosage forms of active compounds by using excipients with low water or low moisture content and manufactured using dry or non-aqueous formulation processes.

[0189] 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 selected from the group consisting of gelatin, hydroxypropyl methylcellulose (HPMC), fish gelatin, and pullulan. In yet another embodiment, the polymeric capsule is selected from the group consisting of gelatin and hydroxypropyl methylcellulose. In yet another embodiment, the polymeric capsule is a hard gelatin capsule.

[0190] 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 yet 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.

[0191] In another embodiment, the pharmaceutical composition further comprises an optional flow aid, and an optional buffering agent. In yet another embodiment, the pharmaceutical composition comprises, as a percentage of total weight, about 0.2% to about 3% by weight of the compound of Formula (I), or a crystalline form thereof, about 86.5% to about 99.8% by weight of a filler, optionally up to about 1.5% by weight of a lubricant, optionally up to about 5% by weight of a flow aid, and optionally up to about 5% by weight of a buffering agent.

[0192] In another embodiment, the pharmaceutical composition comprises, as a percentage of total weight, about 0.2% to about 12% by weight of a compound of formula (I), or a crystalline form thereof, about 76.5% to about 99.8% by weight of a filler, optionally up to about 1.5% by weight of a lubricant, optionally up to about 5% by weight of a flow aid, and optionally up to about 5% by weight of a buffer.

[0193] In some 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 as a percentage of total weight, hi some 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 as a percentage of total weight.

[0194] 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 some 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 yet some other embodiments, the filler is low-moisture microcrystalline cellulose. In some further embodiments, the filler is selected from the group consisting of low-moisture microcrystalline cellulose, sodium starch glycolate, pregelatinized starch, and mixtures thereof.

[0195] In other embodiments, the filler is present in an amount of about 97% to about 99.8% by weight, as a percentage of total weight. In some other embodiments, the filler is present in an amount of about 98% to about 99.75% by weight, as a percentage of total weight. In yet some other embodiments, if a lubricant is present, the amount of filler is reduced by a corresponding percentage of the lubricant present. In some further embodiments, the filler is present in an amount of about 86.5% to about 99.8% by weight, as a percentage of total weight.

[0196] In some embodiments, the filler comprises a first filler and a second filler. The first filler is present in an amount of about 0% to about 99.8% by weight, as a percentage of total weight, and the second filler is present in an amount of about 0% to about 99.8% by weight, as a percentage of total weight, so long as the total weight of the fillers is about 99.8% by weight or less. In some embodiments, the first filler is present in an amount of about 40% to about 60% by weight, as a percentage of total weight, and the second filler is present in an amount of about 40% to about 60% by weight, as a percentage of total weight, so long as the total weight of the fillers is about 99.8% by weight or less.

[0197] In some embodiments, the first filler is low moisture microcrystalline cellulose, starch glycolate, or the like. In some embodiments, the second filler is selected from the group consisting of low-moisture microcrystalline cellulose, sodium starch glycolate, pregelatinized starch, and mixtures thereof.

[0198] Suitable lubricants include, but are not limited to, magnesium stearate, glyceryl behenate, hydrogenated vegetable oil, talc, zinc stearate, calcium stearate, sucrose stearate, sodium stearate fumarate, and mixtures thereof. In some embodiments, the lubricant is magnesium stearate. In other embodiments, the lubricant is present in an amount of about 1.5% by weight as a percentage of the total weight. In yet some other embodiments, the lubricant is present in an amount of about 1% by weight as a percentage of the total weight.

[0199] Suitable flow aids include, but are not limited to, silicon dioxide, talc, and mixtures thereof. In some embodiments, the flow aid is talc. In other embodiments, the flow aid is present in an amount of about 5% by weight as a percentage of the total weight. In some other embodiments, the flow aid is present in an amount of about 1% by weight as a percentage of the total weight. In yet some other embodiments, the flow aid is present in an amount of about 2% by weight as a percentage of the total weight.

[0200] Suitable buffering agents include sodium citrate, citric acid, and mixtures thereof. In some embodiments, the buffering agent is sodium citrate. In some other embodiments, the buffering agent is present in an amount up to about 5% by weight as a percentage of total weight. In yet some other embodiments, the buffering agent is present in an amount up to about 2% by weight as a percentage of total weight.

[0201] In some embodiments, the pharmaceutical composition comprises a compound of formula (I), or a crystalline form thereof, a filler, and optionally a lubricant, wherein: The α-hydroxycarboxylic acid or β-hydroxycarboxylic acid is citric acid, 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 and P is R c -C(O)-, R c -R D and m is 0 or 1, the filler is selected from the group consisting of low moisture microcrystalline cellulose, sodium starch glycolate, pregelatinized starch, and mixtures thereof; The lubricating agent, if present, is magnesium stearate.

[0202] In some embodiments, the pharmaceutical composition comprises a compound of formula (I), or a crystalline form thereof, a filler, and optionally a lubricant, wherein: The compound of formula (I) is (I-1), (I-15), or (I-18), the filler is selected from the group consisting of low moisture microcrystalline cellulose, sodium starch glycolate, pregelatinized starch, and mixtures thereof; The lubricating agent, if present, is magnesium stearate.

[0203] In some embodiments, the pharmaceutical composition comprises about 0.25% to about 2% by weight of a compound of formula (I), or a crystalline form thereof, and about 98% to about 99.75% by weight of a filler, wherein: The compound of formula (I) is (I-1), (I-15), or (I-18), Fillers include low moisture microcrystalline cellulose, sodium starch glycolate, and pregelatinized starches, modified starches, and mixtures thereof.

[0204] In some embodiments, the pharmaceutical composition comprises a compound of formula (I), or a crystalline form thereof, a filler, optionally a lubricant, optionally a flow aid, and optionally a buffer, wherein: The α-hydroxycarboxylic acid or β-hydroxycarboxylic acid is citric acid, A is 0, R ais isobutyl, R a1 is hydrogen, C 1-6 Aliphatic, -(CH2) m -CH2-R B , or -(CH2) m -CH(R 5a )-OR 5b and P is R c -C(O)-, R c -R D and m is 0 or 1, the filler is selected from the group consisting of low moisture microcrystalline cellulose, sodium starch glycolate, pregelatinized starch, and mixtures thereof; The lubricant, if present, is magnesium stearate; the flow aid, when present, is talc; The buffering agent, if present, is sodium citrate.

[0205] In some embodiments, the pharmaceutical composition comprises a compound of formula (I), or a crystalline form thereof, a filler, optionally a lubricant, optionally a flow aid, and optionally a buffer, wherein: The compound of formula (I) is (I-1), (I-15), or (I-18), the filler is selected from the group consisting of low moisture microcrystalline cellulose, sodium starch glycolate, pregelatinized starch, and mixtures thereof; The lubricant, if present, is magnesium stearate; the flow aid, when present, is talc; The buffering agent, if present, is sodium citrate.

[0206] In some embodiments, the pharmaceutical composition comprises, as a percentage of total weight, about 0.2% to about 3% by weight of the compound of Formula (I), or a crystalline form thereof, about 86.5% to about 99.8% by weight of a filler, optionally up to about 1.5% by weight of a lubricant, optionally up to about 5% by weight of a flow aid, and optionally up to about 5% by weight of a buffer, wherein The compound of formula (I) is (I-1), (I-15), or (I-18), the filler is selected from the group consisting of low moisture microcrystalline cellulose, sodium starch glycolate, pregelatinized starch, and mixtures thereof; The lubricant, if present, is magnesium stearate; the flow aid, when present, is talc; The buffering agent, if present, is sodium citrate.

[0207] In some embodiments, a pharmaceutical composition comprises a compound of formula (I), or a crystalline form thereof, a filler, and optionally a lubricant, wherein the compound of formula (I) is (I-1). In some other embodiments, a pharmaceutical composition comprises a compound of formula (I), or a crystalline form thereof, a filler, and optionally a lubricant, wherein the compound of formula (I) is (I-1), the filler is selected from the group consisting of low-moisture microcrystalline cellulose, sodium starch glycolate, pregelatinized starch, and mixtures thereof, and the lubricant, if present, is magnesium stearate.

[0208] In some embodiments, the pharmaceutical composition comprises a compound of formula (I), or a crystalline form thereof, wherein the compound of formula (I) is (I-1) and the crystalline form is Form 2.

[0209] In some embodiments, the pharmaceutical composition comprises the compound of Formula (I-1) Form 2 and a low moisture content. In some other embodiments, the pharmaceutical composition comprises the compound of Formula (I-1) Form 2, and silicified microcrystalline cellulose. In yet some other embodiments, the pharmaceutical composition comprises the compound of Formula (I-1) Form 2, low-moisture microcrystalline cellulose, and magnesium stearate. In still yet some further embodiments, the pharmaceutical composition comprises the compound of Formula (I-1) Form 2, microcrystalline cellulose, and magnesium stearate.

[0210] In some embodiments, the pharmaceutical composition comprises the compound of Formula (I-1) Form 2, low-moisture microcrystalline cellulose, and talc. In some other embodiments, the pharmaceutical composition comprises the compound of Formula (I-1) Form 2 and pregelatinized starch. In still some other embodiments, the pharmaceutical composition comprises the compound of Formula (I-1) Form 2, pregelatinized starch, talc, and magnesium stearate. In still yet some other embodiments, the pharmaceutical composition comprises the compound of Formula (I-1) Form 2, low-moisture microcrystalline cellulose, talc, and magnesium stearate. In some further embodiments, the pharmaceutical composition comprises the compound of Formula (I-1) Form 2, low-moisture microcrystalline cellulose, talc, magnesium stearate, and sodium citrate. In some still further embodiments, the pharmaceutical composition comprises the compound of Formula (I-1) Form 2, low-moisture microcrystalline cellulose, talc, magnesium stearate, and pregelatinized starch. In still yet some further embodiments, the pharmaceutical composition comprises the compound of formula (I-1) Form 2, low-moisture microcrystalline cellulose, talc, magnesium stearate, and sodium starch glycolate.

[0211] When a compound of formula (I) is subjected to hydrolysis conditions, the ester portion of the molecule is hydrolyzed to give a compound of formula (VIII) in a 1:1 molar ratio.

[0212] [ka] Using an analytical method that includes hydrolysis conditions for sample preparation, the amount of compound of Formula (VIII) present in a test sample is determined by comparison to a reference standard of known purity (see, e.g., Analytical Test Method 1 below). Using an analytical method that does not subject the sample to hydrolysis conditions, the amount of compound of Formula (VIII) present in a sample of compound of Formula (I) is determined by comparison to a reference standard of known purity (see, e.g., Analytical Test Method 2 below). Thus, subtracting the amount of compound of Formula (VIII) measured in Analytical Test Method 2 from the amount of compound of Formula (VIII) measured in Analytical Test Method 1 yields the amount of compound of Formula (VIII) in the sample resulting from hydrolysis of compound of Formula (I). Based on a 1:1 molar ratio for the conversion of compound of Formula (I) to compound of Formula (VIII), conversion of molecular weights yields the amount of compound of Formula (I) present in a test sample.

[0213] It will be appreciated that the analytical methods as described immediately above and in the experimental section below can be applied in an analogous manner to any of the compounds of formula (I), (II), (III), (IIIa), (IV), or (IVa), and the various embodiments of these formulas described herein. The analytical methods as described immediately above and in the experimental section below can be applied in an analogous manner to compounds (I-1), (I-15), or (I-18).

[0214] In some embodiments, the amount of the compound of Formula (VIII) present in the pharmaceutical composition is determined by subjecting a sample to conditions under which the compound of Formula (I) is hydrolyzed to the compound of Formula (VIII), and then measuring the amount of the compound of Formula (VIII) present.

[0215] In some embodiments, the amount of the compound of formula (I-1), or a crystalline form thereof, present in the pharmaceutical composition is expressed as an equivalent amount on a molar weight basis of the compound of formula (VIII-1).

[0216] In some embodiments, the present invention relates to a unit dose pharmaceutical composition comprising a compound of formula (I-1), or a crystalline form thereof.

[0217] In some 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 compound of Formula (VIII-1) on a molar weight basis. In still 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 compound of Formula (VIII-1) on a molar weight basis. In still yet 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 compound of Formula (VIII-1) on a molar weight basis. In some further 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 compound of Formula (VIII-1) on a molar weight basis. In yet some further 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 compound of Formula (VIII-1) on a molar weight basis.

[0218] In some embodiments, the amount of the compound of Formula (I-1), or a crystalline form thereof, present in the pharmaceutical composition is expressed as 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).

[0219] In some embodiments, the unit dose pharmaceutical composition contains, on a weight / weight basis, from about 0.143 mg to about 4.3 mg of the compound of Formula (I-1), or a crystalline form thereof, measured as from about 0.1 mg to about 3.0 mg of the compound of Formula (VIII-1).

[0220] In some other embodiments, the unit dose pharmaceutical composition contains, on a weight / weight basis, from about 0.214 mg to about 3.15 mg of the compound of Formula (I-1), or a crystalline form thereof, measured as from about 0.15 mg to about 2.2 mg of the compound of Formula (VIII-1).

[0221] In yet some other embodiments, the unit dose pharmaceutical composition contains, on a weight / weight basis, from about 0.258 mg to about 0.315 mg of the compound of Formula (I-1), or a crystalline form thereof, measured as from about 0.18 mg to about 0.22 mg of the compound of Formula (VIII-1).

[0222] In still yet some other embodiments, the unit dose pharmaceutical composition contains, on a weight / weight basis, from about 0.659 mg to about 0.773 mg of the compound of Formula (I-1), or a crystalline form thereof, measured as from about 0.46 mg to about 0.54 mg of the compound of Formula (VIII-1).

[0223] In some further embodiments, the unit dose pharmaceutical composition comprises, on a weight / weight basis, from about 2.58 mg to about 3.15 mg of the compound of Formula (I-1), or a crystalline form thereof, measured as from about 1.80 mg to about 2.20 mg of the compound of Formula (VIII-1).

[0224] In some embodiments, the present invention provides an oral pharmaceutical preparation of the compound of formula (I), or a crystalline form thereof. A process for manufacturing a dosage form is provided, wherein the oral pharmaceutical dosage form is a capsule; (a-1) mixing together in a bag a sieved filler and a sieved compound of formula (I), or a crystalline form thereof; (a-2) passing the mixture obtained from step (a-1) through a sieve and blending; (a-3) sieving additional filler through the same sieve, passing it through the same bag, and blending it in the same blending device; (a-4) repeating step (a-3) up to two times; (a-5) taking the mixture obtained from step (a-4) and encapsulating it using a capsule filling system; (a-6) weight-classifying the capsules obtained from step (a-5).

[0225] In some embodiments, step (a-3) may be repeated three or more times.

[0226] When a lubricant is present in the pharmaceutical composition, the present invention provides a process for producing an oral pharmaceutical dosage form of the compound of formula (I), or a crystalline form thereof, wherein the oral pharmaceutical dosage form is a capsule; (b-1) mixing together in a bag the sieved filler and the sieved compound of formula (I), or a crystalline form thereof; (b-2) passing the mixture obtained from step (b-1) through a sieve and blending; (b-3) sieving additional filler through the same sieve, passing it through the same bag, and blending it in the same blending device; (b-4) repeating step (b-3) up to two times; (b-5) blending the mixture from step (b-4) with the sieved lubricant; (b-6) taking the mixture obtained from step (b-5) and encapsulating it using a capsule filling system; (b-7) weight-classifying the capsules obtained from step (b-6).

[0227] In some embodiments, step (b-3) may be repeated three or more times. If additional components, such as a buffer, a second bulking agent, or a flow aid, are present in the pharmaceutical composition, they may be added in either step (b-1) or (b-3). The entire amount of each component of the pharmaceutical composition may be added in one step, or may be divided into several amounts, which may or may not be equal amounts, and added to each output of step (b-1) or (b-3).

[0228] In some embodiments, the present invention provides a process for producing an oral pharmaceutical dosage form of the compound of formula (I), or a crystalline form thereof, wherein the oral pharmaceutical dosage form is a capsule; (c-1) passing a filler through a sieve and placing it in a high shear mixer; (c-2) passing the compound of formula (I), or a crystalline form thereof, through a sieve and placing it in the same high shear mixing device; (c-3) passing the filler through a sieve and placing it in the same high shear mixer; (c-4) mixing for less than 10 minutes using the same high shear mixer; (c-5) taking the mixture obtained from step (c-4) and encapsulating it using a capsule filling system; and (c-6) encapsulating the mixture obtained from step (c-5). and weight sorting the capsules.

[0229] In some embodiments, when a high shear mixing device is used, additional ingredients present in the pharmaceutical composition may be added by repeating either step (c-1) or step (c-3).

[0230] In some embodiments, the compound of formula (I) used in the process for preparing the solid oral dosage form described above is selected from the group consisting of (I-1), (I-15), and (I-18). In some embodiments, the compound of formula (I) used in the process for preparing the solid oral dosage form described above is (I-1).

[0231] The process outlined above can be carried out using conventional equipment and facilities. For a review, see, for example, Remington: The Science and Practice of Pharmacy, 21 st Ed., Lippincott Williams & Wilkins, 2005.

[0232] The blending steps outlined above can be performed using any conventional blending equipment. In some embodiments, the blending time for each individual blending step is from about 1 minute to about 45 minutes. In some other embodiments, the blending time for each individual blending step is from about 1 minute to about 20 minutes. In yet some other embodiments, the blending time for each individual blending step is from about 2 minutes to about 15 minutes.

[0233] The mixing step outlined above can be carried out in any conventional polyethylene bag. In some embodiments, the mixing step is for about 30 seconds to 5 minutes. In some embodiments, the mixing step outlined above can be carried out in a stainless steel container.

[0234] The mixing step using a high shear mixer can be performed with any conventional high shear mixer. One example of such a high shear mixer is sold as the Lab High Shear Granulator (Key International, Inc., Englishtown, NJ). In some embodiments, mixing is performed for less than about 10 minutes. In some other embodiments, mixing is performed for less than about 5 minutes.

[0235] The capsule filling steps outlined above can be performed with any conventional capsule filling system or equipment. In some embodiments, the capsule filling step is semi-automated and capable of processing small batch sizes. An example of such a capsule filling system is sold as In-Cap (Isopak Limited, Lincolnshire, Stamford, United Kingdom). In some embodiments, the capsule filling system is manual. An example of such a capsule filling device is sold as ProFill 100 (Torpac, Inc., Fairfield, NJ, USA).

[0236] In some embodiments, the capsules are hard gelatin capsules, sold as Coni-Snap® (Capsugel, Peapack, NJ). One of ordinary skill in the art would be able to select an appropriate capsule size and color. In some embodiments, the capsules have a fill weight of 85 mg, 120 mg, or 150 mg.

[0237] The weight sorting step outlined above can be performed using any conventional weight sorter or machine. One example of a weight sorter or machine is sold as the SADE SP Bench Top Tablet and Capsule Weight Sorter (AC Compacting LLC, North Brunswick, NJ, USA). There are.

[0238] In some embodiments, the capsules are packaged in a bottle, an aluminum foil pouch, or a blister pack. In some other embodiments, the capsules are packaged in a high-density polyethylene (HDPE) bottle sealed with a heat induction seal. In another embodiment, the capsules are packaged in a hermetically sealed aluminum foil pouch. In yet another embodiment, the capsules are packaged in a foil-foil blister pack. In some other embodiments, the capsules are packaged with a desiccant.

[0239] The physical and chemical stability of the oral pharmaceutical dosage form may be tested by conventional methods, for example, measuring dissolution, disintegration time, assaying for degradation products of the compound of formula (I) after storage at different temperatures for different lengths of time.

[0240] In some other embodiments, the present invention provides pharmaceutical compositions for parenteral use. In yet some other embodiments, the present invention provides liquid pharmaceutical compositions for parenteral or oral use.

[0241] In some embodiments, the compound of formula (I) is formulated as a lyophilized powder in a manner similar to that described in Plamondon et al., WO 02 / 059131, incorporated herein by reference in its entirety. In such embodiments, an aqueous mixture comprising an α-hydroxycarboxylic acid or a β-hydroxycarboxylic acid is hydrolyzed to form the compound of formula (I).

[0242] In some embodiments, the lyophilized powder also contains a free α-hydroxycarboxylic acid or β-hydroxycarboxylic acid. Preferably, the free α-hydroxycarboxylic acid or β-hydroxycarboxylic acid compound and the compound of Formula (I) are present in the mixture in a molar ratio ranging from about 0.5:1 to about 100:1, more preferably from about 5:1 to about 100:1. In various embodiments, the α-hydroxycarboxylic acid or β-hydroxycarboxylic acid compound is citric acid, and the lyophilized powder contains the free citric acid and the corresponding boronic acid ester in a molar ratio ranging from about 10:1 to about 100:1, from about 20:1 to about 100:1, or from about 40:1 to about 100:1.

[0243] In some embodiments, the lyophilized powder contains citric acid and a compound of Formula (I), and is substantially free of other ingredients. However, the composition can further contain one or more other pharmaceutically acceptable excipients, carriers, fillers, salts, buffers, bulking agents, stabilizers, solubilizers, and other substances known in the art. Preparation of pharmaceutically acceptable formulations containing these substances is described, for example, in Remington: The Science and Practice of Pharmacy, 20th Ed., ed. A. Gennaro, Lippincott Williams & Wilkins, 2000, or the latest edition, and Strickley, Pharmaceutical Research, 21(2)201-230(2004).

[0244] Upon dissolution in aqueous medium, equilibrium is established between the boronic ester compound of Formula (I) and the corresponding free boronic acid compound. In some embodiments, equilibrium is achieved rapidly, for example, within 1 to 15 minutes after addition of aqueous medium. The relative concentrations of the boronic ester, boronic acid, and any intermediate species present at equilibrium will vary depending on parameters such as solution pH, temperature, the nature of the α-hydroxycarboxylic acid or β-hydroxycarboxylic acid, and the ratio of boronic ester of α-hydroxycarboxylic acid or β-hydroxycarboxylic acid to the compound of Formula (I) present in the lyophilized powder.

[0245] In some embodiments, the pharmaceutical composition comprises a compound of formula (I), a bulking agent, and a buffering agent. In some other embodiments, the pharmaceutical composition comprises a compound of formula (I), a bulking agent, and a buffering agent in a lyophilized powder.

[0246] In some embodiments, the compound of formula (I) is preformed. In some other embodiments, the compound of formula (I) is formed in situ from the corresponding boronic acid of formula (VIII). In yet some other embodiments, compound (I-1) is preformed. In still yet some other embodiments, compound (I-15) is formed in situ from compound (VIII-15).

[0247] A suitable bulking agent includes glycine. In some embodiments, the bulking agent is present in an amount of about 1% weight / volume (w / v) to about 5% w / v. In some other embodiments, the bulking agent is present in an amount of about 3% w / v.

[0248] Suitable buffering agents include sodium citrate, citric acid, and mixtures thereof, hi some embodiments, the buffering agent is sodium citrate and citric acid.

[0249] In some embodiments, the buffering agent is present at a concentration of about 45 mM to about 65 mM, hi some other embodiments, the buffering agent is present at a concentration of about 50 mM to about 60 mM.

[0250] In some embodiments, the ratio of buffering agent to compound of Formula (I) is about 50:1 to about 10:1. In some other embodiments, the ratio of buffering agent to compound of Formula (I) is about 30:1 to about 10:1. In yet some other embodiments, the ratio of buffering agent to compound of Formula (I) is about 20:1.

[0251] In some embodiments, the pH of the pharmaceutical composition is about pH 4.7 to pH 6.1. The pH of the pharmaceutical composition can be adjusted using any suitable inorganic or organic acid.

[0252] In some embodiments, the pharmaceutical composition comprises a compound of formula (I), a bulking agent, and a buffering agent, wherein: The α-hydroxycarboxylic acid or β-hydroxycarboxylic acid is citric acid, A is 0, R ais isobutyl, R a1 is hydrogen, C 1-6 Aliphatic, -(CH2) m -CH2-R B , or -(CH2) m -CH(R 5a )-OR 5b and P is R c -C(O)-, R c -R D and m is 0 or 1, The bulking agent is glycine, The buffering agents are sodium citrate and citric acid.

[0253] In some embodiments, the pharmaceutical composition comprises a compound of formula (I), a bulking agent, and a buffering agent, wherein: The compound of formula (I) is represented by compound (I-1), (I-15), or (I-18), The bulking agent is glycine, The buffering agents are sodium citrate and citric acid.

[0254] In some embodiments, the pharmaceutical composition comprises a lyophilized powder of the compound of formula (I), a bulking agent, and a buffering agent, wherein: The α-hydroxycarboxylic acid or β-hydroxycarboxylic acid is citric acid, 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 and P is R c -C(O)-, R c -R D and m is 0 or 1, The bulking agent is glycine, The buffering agents are sodium citrate and citric acid.

[0255] In some embodiments, the pharmaceutical composition comprises a lyophilized powder of the compound of formula (I), a bulking agent, and a buffering agent, wherein: The compound of formula (I) is represented by compound (I-1), (I-15), or (I-18), The bulking agent is glycine, The buffering agents are sodium citrate and citric acid.

[0256] In some embodiments, the pharmaceutical composition is a lyophilized powder of compound (I-1). In some other embodiments, the pharmaceutical composition comprises a lyophilized powder of compound (I-1), glycine, sodium citrate, and citric acid. In yet some other embodiments, the pharmaceutical composition comprises a lyophilized powder of compound (I-15). In still yet some other embodiments, the pharmaceutical composition comprises a lyophilized powder of compound (I-15), glycine, sodium citrate, and citric acid.

[0257] In some embodiments, the present invention provides a unit dose pharmaceutical composition comprising a lyophilized powder of the compound of formula (I-1), a bulking agent, and a buffering agent. In some embodiments, the unit dose pharmaceutical composition comprises a lyophilized powder of the compound of formula (I-1), glycine, sodium citrate, and citric acid.

[0258] In some embodiments, the compound of Formula (I-1) is present in the unit dose pharmaceutical composition in an amount equivalent to about 1 mg to about 10 mg of the compound of Formula (VIII-1) by molar weight. In some embodiments, the compound of Formula (I-1) is present in the unit dose pharmaceutical composition in an amount equivalent to about 1 mg to about 5 mg of the compound of Formula (VIII-1) by molar weight. In some embodiments, the compound of Formula (I-1) 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 5.0 mg of the compound of Formula (VIII-1) by molar weight. In some embodiments, the compound of Formula (I-1) is present in the unit dose pharmaceutical composition in an amount equivalent to about 3.5 mg of the compound of Formula (VIII-1) by molar weight.

[0259] In some embodiments, the amount of glycine present in a unit dose pharmaceutical composition is about 0.01 g to about 0.50 g. In some embodiments, the amount of glycine present in a unit dose pharmaceutical composition is about 0.03 g to about 0.250 g. In some embodiments, the amount of glycine present in a unit dose pharmaceutical composition is about 0.06 g to about 0.125 g.

[0260] In some embodiments, the 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, hi some embodiments, the 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.

[0261] In some embodiments, the present invention provides a lyophilized powder of a compound of formula (I-15), a bulking agent, and a buffering agent. In some embodiments, the unit dose pharmaceutical composition comprises a lyophilized powder of the compound of formula (I-15), glycine, sodium citrate, and citric acid.

[0262] In some 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 compound of Formula (VIII-15) by molar weight. In some 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 compound of Formula (VIII-15) by molar weight. In some 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 5.0 mg of the compound of Formula (VIII-15) by molar weight. In some 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 compound of Formula (VIII-15) by molar weight.

[0263] In some embodiments, the amount of glycine present in a unit dose pharmaceutical composition is about 0.01 g to about 0.50 g. In some embodiments, the amount of glycine present in a unit dose pharmaceutical composition is about 0.03 g to about 0.250 g. In some embodiments, the amount of glycine present in a unit dose pharmaceutical composition is about 0.06 g to about 0.125 g.

[0264] In some embodiments, the 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, hi some embodiments, the 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.

[0265] In another aspect, the present invention provides a process for preparing a compound of formula (I) as a lyophilized powder, the process comprising: (d-1) i. aqueous solvent mixture, ii. A compound of formula (I), iii. bulking agents, and iv. mixing a buffer to form a mixture; (d-2) freeze-drying the mixture.

[0266] In some embodiments, the compound of formula (I) is formed in situ from the corresponding compound of formula (VIII). Thus, the present invention also provides a method for preparing a compound of formula (I) as a lyophilized powder, the method comprising: (e-1) i. aqueous solvent mixture, ii. A compound of formula (VIII): iii. bulking agents, and iv. mixing an α-hydroxycarboxylic acid or a salt thereof, or a β-hydroxycarboxylic acid or a salt thereof to form a mixture; (e-2) freeze-drying the mixture.

[0267] In some embodiments, the aqueous solvent mixture comprises one or more co-solvents in addition to water. In some embodiments, the co-solvent is miscible with water. In some other embodiments, the co-solvent is an alcohol, including, but not limited to, ethanol, tert-butyl alcohol, and mixtures thereof. In some other embodiments, the co-solvent is tert-butyl alcohol.

[0268] In some embodiments, the aqueous solvent mixture comprises between about 1% v / v and about 40% v / v alcohol. In some other embodiments, the aqueous solvent mixture comprises between about 3% v / v and about 10% v / v alcohol. In some other embodiments, the aqueous solvent mixture comprises between about 3% v / v and about 6% v / v alcohol. In yet some other embodiments, the solvent mixture comprises between about 3% v / v and about 6% v / v tert-butyl alcohol. In still yet some other embodiments, the solvent mixture comprises about 5% v / v tert-butyl alcohol.

[0269] In some embodiments, a method for preparing compound (I-1) as a lyophilized powder is provided, the method comprising: (f-1) i. Water, ii. Compound (I-1), iii. glycine, iv. sodium citrate, and v. citric acid to form a mixture; (f-2) freeze-drying the mixture.

[0270] In some embodiments, a method for preparing compound (I-15) as a lyophilized powder is provided, the method comprising: (g-1) i. an aqueous solvent mixture comprising water and tert-butyl alcohol; ii. Compound (VIII-15), iii. glycine, iv. sodium citrate, and v. citric acid to form a mixture; (g-2) freeze-drying the mixture.

[0271] In some other embodiments, for the method described immediately above, the amount of tert-butyl alcohol present in the aqueous solvent mixture is from about 3% v / v to about 6% v / v.

[0272] Lyophilization or freeze-drying can be carried out using any conventional freeze-drying machine or freeze-dryer. In some embodiments, freeze-drying includes (i) loading and freezing the liquid mixture as prepared above, (ii) annealing, (iii) a second freezing cycle, (iv) drying under vacuum, and (v) secondary drying. The temperature and time of each step can vary depending on the freeze-drying machine or freeze-dryer used.

[0273] In some embodiments, the resulting lyophilized powder has a residual moisture content of less than about 2%. In some other embodiments, the resulting lyophilized powder has a residual moisture content of less than about 1%.

[0274] In another aspect, the present invention provides a method for preparing a pharmaceutical composition of a compound of Formula (I) as a liquid pharmaceutical dosage form, the method comprising 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, saline, phosphate-buffered saline (PBS), and mixtures thereof. In some embodiments, the reconstitution solvent is water, water for injection, saline, and mixtures thereof. In some other embodiments, the reconstitution solvent is water for injection. After reconstitution, the liquid pharmaceutical dosage form can contain the concentration of the compound of Formula (I) described herein.

[0275] In some embodiments, a method for preparing a pharmaceutical composition of Compound (I) as a liquid pharmaceutical dosage form is provided, the method comprising reconstituting a lyophilized powder of Compound (I) described herein with an aqueous solvent suitable for pharmaceutical administration. In some embodiments, a method for preparing a pharmaceutical composition of Compound (I-1) as a liquid pharmaceutical dosage form is provided, the method comprising reconstituting a lyophilized powder of Compound (I-1) described herein with water for injection or saline. In some embodiments, a method for preparing a pharmaceutical composition of Compound (I-1) as a liquid pharmaceutical dosage form is provided, the method comprising reconstituting a lyophilized powder of Compound (I-1) described herein with water for injection.

[0276] In some embodiments, a method for preparing a pharmaceutical composition of compound (I-15) as a liquid pharmaceutical dosage form is provided, the method comprising reconstituting a lyophilized powder of compound (I-15) described herein with an aqueous solvent suitable for pharmaceutical administration. In some embodiments, a method for preparing a pharmaceutical composition of compound (I-15) as a liquid pharmaceutical dosage form is provided, the method comprising reconstituting a lyophilized powder of compound (I-15) described herein with water for injection or saline. In some embodiments, a method for preparing a pharmaceutical composition of compound (I-15) as a liquid pharmaceutical dosage form is provided, the method comprising reconstituting a lyophilized powder of compound (I-15) described herein with water for injection.

[0277] Upon reconstitution in the reconstitution solvent, equilibrium is established between the compound of Formula (I) and the corresponding boronic acid of Formula (VIII). Equilibrium is generally achieved rapidly, within approximately 10-15 minutes after addition of the reconstitution solvent. The relative concentrations of boronic ester and boronic acid present at equilibrium vary depending on the solution's pH, temperature, and the ratio of α-hydroxycarboxylic acid or β-hydroxycarboxylic acid to boronic acid.

[0278] In another aspect, the present invention provides a liquid pharmaceutical composition comprising a compound of Formula (I) and additional excipients as described herein. In some embodiments, the liquid pharmaceutical composition is suitable for parenteral administration. In some other embodiments, the liquid pharmaceutical composition is suitable for oral administration.

[0279] In such embodiments, the liquid pharmaceutical composition comprises a compound of formula (I), a buffering agent, and optionally a tonicity adjusting agent.

[0280] In some embodiments, the ratio of buffering agent to compound of Formula (I) is about 50:1 to about 10:1. In some other embodiments, the ratio of buffering agent to compound of Formula (I) is about 30:1 to about 10:1. In yet some other embodiments, the ratio of buffering agent to compound of Formula (I) is about 20:1.

[0281] In some embodiments, the buffering agent is present at a concentration of about 45 mM to about 65 mM, hi some other embodiments, the buffering agent is present at a concentration of about 50 mM to about 60 mM.

[0282] Suitable buffering agents include sodium citrate, citric acid, and mixtures thereof, hi some embodiments, the buffering agent is sodium citrate and citric acid.

[0283] 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 some embodiments, the tonicity adjusting agent is propylene glycol.In some other embodiments, the tonicity adjusting agent is sodium chloride.

[0284] Upon dissolution in the aqueous solvent mixture, equilibrium is established between the compound of Formula (I) and the corresponding boronic acid of Formula (VIII). Therefore, either the compound of Formula (I) or the compound of Formula (VIII) can be used in the preparation of liquid pharmaceutical compositions. Generally, equilibrium is achieved rapidly, within about 10 to 15 minutes after addition of the aqueous solvent mixture. The relative concentrations of the boronic ester and boronic acid present at equilibrium vary depending on the solution's pH, temperature, and the ratio of α-hydroxycarboxylic acid or β-hydroxycarboxylic acid compound to boronic acid compound. In some embodiments, excess α-hydroxy or β-hydroxy acid can act as a stabilizer, driving the boronic ester toward equilibrium. In some embodiments, a tonicity modifier can also act as a stabilizer.

[0285] In some embodiments, the liquid pharmaceutical composition optionally further comprises a preservative.

[0286] In some embodiments, the liquid pharmaceutical composition comprises a compound of formula (I), a buffering agent, and optionally, a tonicity adjusting agent, wherein: The α-hydroxycarboxylic acid or β-hydroxycarboxylic acid is citric acid, 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 and P is R c -C(O)-, R c -R D and m is 0 or 1, The buffering agents are sodium citrate and citric acid, The tonicity adjusting agent, if present, is sodium chloride.

[0287] In some embodiments, the liquid pharmaceutical composition comprises a compound of formula (I), a buffering agent, and optionally, a tonicity adjusting agent, wherein: The compound of formula (I) is represented by compound (I-1), (I-15), or (I-18), and the buffer is sodium citrate and citric acid; The tonicity adjusting agent, if present, is sodium chloride.

[0288] In some embodiments, the α-hydroxycarboxylic acid or β-hydroxycarboxylic acid is citric acid, and the liquid pharmaceutical composition of the compound of Formula (I) comprises the compound of Formula (I), water, citric acid, sodium citrate, and sodium chloride. In some other embodiments, the α-hydroxycarboxylic acid or β-hydroxycarboxylic acid is citric acid, and the liquid pharmaceutical composition comprises the compound of Formula (I), water, citric acid, and propylene glycol. In yet some other embodiments, the liquid pharmaceutical composition comprises the compound of Formula (I), and the compound of Formula (I) is Compound (I-1), water, citric acid, sodium citrate, and sodium chloride.

[0289] In such embodiments, the α-hydroxycarboxylic acid or β-hydroxycarboxylic acid is citric acid, and the liquid pharmaceutical dosage form of the compound of Formula (I) has a pH of about pH 3 to about pH 7. In certain such embodiments, the pH is about pH 4.9 to about pH 6.7. In other certain such embodiments, the pH is about pH 5.5 to about pH 6.5.

[0290] In some embodiments, the α-hydroxycarboxylic acid or β-hydroxycarboxylic acid is citric acid, and the liquid pharmaceutical composition of the compound of Formula (I) is prepared in situ from a storage vehicle and a compound of Formula (VIII). In some embodiments, the storage vehicle comprises water, citric acid, sodium citrate, and propylene glycol. In such embodiments, the resulting solution is further diluted with the storage vehicle or sodium chloride solution, A liquid pharmaceutical composition of a compound of formula (I) at a desired concentration can be produced.

[0291] In another aspect, the present invention provides a unit-dose liquid pharmaceutical composition comprising a compound of Formula (I), a buffer, and optionally a tonicity adjusting agent. In some 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 some 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 some 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 some other embodiments, the compound of Formula (I) is compound (I-1), and in the unit-dose liquid pharmaceutical composition, compound (I-1) is present in a concentration of about 0.5 mg / mL to about 3 mg / mL of the compound of Formula (VIII-1). In still some other embodiments, the compound of Formula (I) is compound (I-1), and in the unit-dose liquid pharmaceutical composition, compound (I-1) is present in a compound of Formula (VIII-1) at a concentration of about 1 mg / mL. In still still some other embodiments, the compound of Formula (I) is compound (I-15), and in the unit-dose liquid pharmaceutical composition, compound (I-15) is present in a compound of Formula (VIII-15) at a concentration of about 1 mg / mL.

[0292] In some embodiments, in a unit dose liquid pharmaceutical composition, sodium citrate and citric acid are present in an amount equivalent to about 0.005 g to about 0.250 g of citrate ion. In some embodiments, in a unit dose liquid pharmaceutical composition, sodium citrate and citric acid are present in an amount equivalent to about 0.025 g to about 0.125 g of citrate ion.

[0293] In some embodiments, sodium chloride is present in a unit dose liquid pharmaceutical composition in an amount of about 0.0045 g to about 0.09 g. In some embodiments, sodium chloride is present in a unit dose liquid pharmaceutical composition in an amount of about 0.01 g to about 0.04 g.

[0294] In some embodiments of unit dose liquid pharmaceutical compositions, the pharmaceutical composition is stored frozen until use.

[0295] In another aspect, the present invention provides a method for preparing a compound of formula (I) as a unit dose liquid pharmaceutical composition, the method comprising: (h-1) dissolving a buffer in an aqueous solvent; (h-2) dissolving the compound of formula (I), or a crystalline form thereof, in the mixture obtained in step (h-1); (h-3) dissolving a tonicity adjusting agent in the mixture obtained in step (h-2); (h-4) adding additional aqueous solvent to the required batch volume; (h-5) filling a vial with the amount of the mixture obtained in step (h-4).

[0296] In some embodiments, after step (h-5), the vial is stoppered. In some other embodiments, nitrogen is bubbled through the mixture before step (h-5). In yet some other embodiments, after step (h-5), the liquid in the vial can be overlaid with nitrogen.

[0297] In some embodiments, the compound of formula (I) is formed in situ from the compound of formula (VIII). In such embodiments, in step (h-2), the compound of formula (VIII), or a crystalline form thereof, is added to the mixture. In some embodiments, an alpha-hydroxy acid or a beta-hydroxy acid is added in step (h-2). In some other embodiments, In embodiments, an alpha-hydroxy acid or a beta-hydroxy acid is present in step (h-1) as a buffering agent.

[0298] The pharmaceutical compositions of the present invention are preferably formulated for administration to patients suffering from or at risk of developing or experiencing a recurrence of a proteasome-mediated disease. The term "patient," as used herein, refers to an animal, preferably a mammal, and more preferably a human. Preferred pharmaceutical compositions of the present invention are formulated for oral, intravenous, or subcutaneous administration. However, any of the above dosage forms containing a therapeutically effective amount of a compound of the present invention is well within the bounds of routine experimentation and therefore well within the scope of the present invention. In some embodiments, the pharmaceutical compositions of the present invention may further comprise another therapeutic agent. In some embodiments, such other therapeutic agent is one that is normally administered to patients with the disease or condition to be treated.

[0299] A "therapeutically effective amount" refers to an amount sufficient to produce a detectable decrease in proteasome activity or the severity of a proteasome-mediated disease. The amount of proteasome inhibitor required may vary depending on the effectiveness of the inhibitor for a given cell type and the time required to treat the disease. It should also be understood that the specific drug and treatment regimen required for any particular patient will depend on a variety of factors, including the activity of the specific compound used, age, weight, overall health, sex, diet of the patient, time of administration, excretion rate, concomitant medications, 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 the present invention is generally less than the amount normally administered in a composition containing that therapeutic agent as the sole active agent. Preferably, the amount of additional therapeutic agent can range from about 50% to about 100% of the amount normally present in a composition containing that agent as the sole therapeutic active agent.

[0300] In another aspect, the present invention provides methods for treating a patient suffering from or at risk of developing or experiencing a recurrence of a proteasome-mediated disease. As used herein, the term "proteasome-mediated disease" includes any disease, disorder, or condition caused by or characterized by increased proteasome expression or activity, or in which proteasome activity is required. The term "proteasome-mediated disease" also includes any disease, disorder, or condition in which inhibition of proteasome activity is beneficial.

[0301] For example, the compounds and pharmaceutical compositions of the present invention may inhibit the activity of proteins (e.g., NFκB, p27 Kip , p21 WAF / CIP1 , p53). Related diseases include inflammatory diseases (e.g., rheumatoid arthritis, inflammatory bowel disease, asthma, chronic obstructive pulmonary disease (COPD), osteoarthritis, skin diseases (e.g., atopic dermatitis, psoriasis)), vascular proliferative diseases (e.g., atherosclerosis, restenosis), proliferative eye diseases (e.g., diabetic retinopathy), benign proliferative disorders (e.g., hemangiomatosis), autoimmune diseases (e.g., multiple sclerosis, tissue and organ rejection), and inflammation associated with infection (e.g., immune reaction). These include neurodegenerative disorders (e.g., neurodegeneration as a result of Alzheimer's disease, Parkinson's disease, motor neuron disease, neuropathic pain, triplet repeat disease, astrocytoma, and alcoholic liver disease), ischemic injury (e.g., cerebral infarction), and cachexia (e.g., accelerated muscle protein breakdown that accompanies various physiological and pathological conditions, such as neuronal injury, fasting, fever, acidosis, HIV infection, cancer afflictions, and certain endocrine disorders).

[0302] The compounds and pharmaceutical compositions of the present invention are particularly useful for treating cancer. As used herein, the term "cancer" refers to a cancer characterized by uncontrolled or unregulated cell proliferation, decreased cell differentiation, inappropriate ability to invade surrounding tissues, and / or new growth at ectopic sites. "Cancer" refers to a cellular disorder characterized by the ability to form new cells. 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 includes primary and metastatic cancers.

[0303] Non-limiting examples of solid tumors treatable 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, e.g., metastatic renal cell carcinoma), hepatocellular carcinoma, lung cancer (including, e.g., non-small cell lung cancer (NSCLC) and bronchioloalveolar carcinoma (BAC)), ovarian cancer (including, e.g., advanced epithelial carcinoma or primary peritoneal carcinoma), cervical cancer, gastric cancer, esophageal cancer, head and neck cancer (including, e.g., squamous cell carcinoma of the head and neck), melanoma, neuroendocrine carcinoma, including metastatic neuroendocrine tumors, brain tumors (including, e.g., glioma, anaplastic oligodendroglioma, glioblastoma multiforme, and adult anaplastic astrocytoma), osteosarcoma, and sarcoma of soft tissue.

[0304] Non-limiting examples of hematological malignancies treatable with the disclosed proteasome inhibitors or pharmaceutical compositions include acute myeloid leukemia (AML), chronic myeloid leukemia (CML), including CML accelerated phase 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), myelodysplastic syndromes (MDS), including Waldenstrom's macroglobulinemia, refractory anemia (RA), refractory anemia with ringed sideroblasts (RARS), refractory anemia with excess blasts (RAEB), and accelerated RAEB (RAEB-T), and myeloproliferative syndromes.

[0305] In some embodiments, the compounds or pharmaceutical compositions of the present invention are used to treat a patient suffering from, or experiencing the onset or recurrence of, a cancer selected from the group consisting of multiple myeloma and mantle cell lymphoma.

[0306] In some embodiments, the proteasome inhibitor or pharmaceutical composition of the present invention is administered in combination with another therapeutic agent. The other therapeutic agent may also inhibit the proteasome or may act by a different mechanism. In some embodiments, the other therapeutic agent is one that is normally administered to patients with the disease or condition to be treated. The proteasome inhibitor of the present invention may be administered together with the other therapeutic agent in a single dosage form or as a separate dosage form. When administered as a separate dosage form, the other therapeutic agent may be administered before, simultaneously with, or after administering the proteasome inhibitor of the present invention.

[0307] In some embodiments, the proteasome inhibitor of Formula (I) or the pharmaceutical composition of the compound of Formula (I) is administered in combination with an anti-cancer agent. As used herein, the term "anti-cancer agent" refers to any agent administered to a subject with cancer for the purpose of treating the cancer.

[0308] Non-limiting examples of DNA damaging chemotherapeutic agents include topoisomerase I inhibitors (e.g., irinotecan, topotecan, camptothecin, or its analogs, and doxorubicin), topoisomerase II inhibitors (e.g., etoposide, teniposide, and daunorubicin), alkylating agents (e.g., melphalan, chlorambucil, busulfan, thiotepa, ifosfamide, carmustine, lomustine, semustine, streptozocin, dacarbazine, 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, capecitabine, gemcitabine, fludarabine, , cytarabine, mercaptopurine, thioguanine, pentostatin, and hydroxyurea).

[0309] Chemotherapeutic agents that interfere with cell replication include paclitaxel, docetaxel and related analogs, vincristine, vinblastine 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 inhibitors of IκB kinase, antibodies that bind to proteins overexpressed in cancer and thereby downregulate cell replication (e.g., trastuzumab, rituximab, cetuximab, and bevacizumab), and other inhibitors of proteins or enzymes known to be upregulated, overexpressed, or activated in cancer, inhibitors that downregulate cell replication.

[0310] In order that this invention may be more fully understood, the following preparative and experimental examples are set forth which illustrate how to make or test particular compounds and are not to be construed as limiting the scope of the invention in any way. [Example]

[0311] Example

[0312]

number

[0313] Mass spectrometry: Mass spectrometry experiments were performed on a Thermo-Finnigan LCQ Deca-XP ion trap mass spectrometer. The electrospray ion source was used in both positive and negative modes with 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 offset of 35 V. Analytes were dissolved in acetonitrile to produce 0.5 mg / mL solutions. An Agilent 1100 HPLC system was used for liquid chromatography-mass spectrometry flow analysis. The pump flow rate was 1.0 mL / min. 10 μL of each sample solution was injected into a T-joint via an autosampler. Approximately 2% of the solution from the T-joint was injected into the mass spectrometer.

[0314] X-ray Powder Diffraction (XRPD): X-ray powder diffraction patterns are obtained on either: i) a Bruker AXS D8Advance diffractometer. Data are collected over the angular range of 2.9° to 29.6° 2θ in continuous scan mode using a 0.05° 2θ step size and a 2 second step time. Samples are operated under ambient conditions and prepared as flat plate samples using powders if accepted without grinding, or ii) PANalytical X'Pert Pro Diffractometer. Each sample was analyzed using copper radiation generated with a long, fine-focus source manufactured by Optix. An elliptically graded multilayer mirror was used to focus the Cu Kα X-ray source through the sample onto the detector. The sample was analyzed in transmission geometry, sandwiched between 3-micrometer thick films, and rotated to optimize orientation statistics. A beam stop was used to minimize background generated by air scattering. Helium and anti-scatter extensions were not used. Soller slits were used on the incident and diffracted beams to minimize axial divergence. Diffraction patterns were collected with axial divergence minimized for the scan position. Diffraction patterns were collected with a scanning position-sensitive detector (X'Celerator), positioned 240 mm from the sample. Prior to analysis, a silicon sample (NIST Standard Reference Material 640c) was analyzed to verify the position of the silicon-111 peak.

[0315] Differential Scanning Calorimetry (DSC): Differential scanning calorimetry (DSC) data are collected either i) TA Instruments Q100 Differential Scanning Calorimeter equipped with a 50-position autosampler. The energy and temperature calibration standard is indium. Samples are heated from 25°C to 300°C at a rate of 10°C per minute. A nitrogen purge flowing at 50 mL per minute is maintained over the sample during the scan. Samples of 1 mg to 3 mg are analyzed. All samples are crimped into hermetically sealed aluminum pans with pinholes to relieve pressure buildup from solvent vapors or ii) TA Instruments Differential Scanning Calorimeter 2920. The sample is placed in an aluminum DSC pan and the weight is accurately recorded. The open pan is covered with a lid and then crimped. The sample cell is equilibrated at 25°C and heated at a rate of 10°C / min under a nitrogen purge. Indium metal was used as the calibration standard.

[0316] Thermogravimetric Analysis (TGA): Thermogravimetric analysis (TGA) data are collected on a TA Instruments Q500 thermogravimetric analyzer calibrated with nickel / alumel, with runs at a scan rate of 10°C per minute. A nitrogen purge flowing at 60 mL per minute is passed over the sample during the measurements. Typically, 5 mg to 15 mg of sample was placed in a pre-tared platinum crucible.

[0317] Example 1: Synthesis of 4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-methylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid (I-1)

[0318] [ka] Step 1: 2,5-[(dichlorobenzoyl)amino]acetic acid To a mixture of NaOH (12 g, 300 mmol) and glycine (18 g, 239 mmol) in water (120 mL) was added dropwise over 45 minutes a solution of 2,5-dichlorobenzoyl chloride (10 g, 48 mmol) in THF (15 mL), maintaining the internal temperature below approximately 25° C. After 1 hour, the mixture was acidified with 2.0 M HCl (125 mL), maintaining the internal temperature below approximately 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%). Melting point: 173.3° C. 1 H NMR (300MHz, DMSO-d6, δ): 12.72 (bs, 1H), 8.89 (t, J = 6.0 Hz, 1 H), 7.54 (m, 2 H), 7.48 (m, 1 H), 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] calculated for C9H8Cl2NO3 248.0; found 248.0; [M+Na] calculated for C9H7Cl2NNaO3 270.0; found 270.2.

[0319] 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), 2.0 M NaOH (130 mL) was added, 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 a rate such that the internal temperature was maintained at 0±1°C. During the addition, the pH was controlled at 11.0±0.2 using a pH controller titrated with 2.0 M NaOH. After the addition was complete, the mixture was stirred for an additional 2 hours at 0±1°C. The mixture was then acidified with 2.0 M HCl (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 (57.6 g, 97.3%) as a white solid.

[0320] Step 2: 2,5-Dichloro-N-[2-({(1R)-3-methyl-1-[(3a S,4S,6S,7aR)-3a,5,5-trimethylhexahydro-4,6-methano-1,3,2-benzodioxaborol-2-yl]butyl}amino)-2-oxoethyl]benzamide To a solution of 2,5-[(dichlorobenzoyl)amino]acetic acid (6.10 g, 24.6 mmol) and TBTU (8.34 g, 26.0 mmol) in DMF (40 mL) with an internal temperature below about 5° C., (1R)-3-methyl-1-[(3aS,4S,6S,7aR)-3a,5,5-trimethylhexahydro-4,6-methano-1,3,2-benzodioxaborol-2-yl]butan-1-amine·TFA (9.35 g, 24.7 mmol) was added. DIPEA (13 mL, 75 mmol) was then added dropwise over 2 h, maintaining the internal temperature below about 5° C. After 40 min, the mixture was diluted with EtOAc (90 mL) and washed with 5% NaCl (150 mL), twice with 10% NaCl (2 × 40 mL), once with 2% KCO (1 × 40 mL), once with 1% HPO (1 × 40 mL), and once with 10% NaCl (1 × 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-benzodioxaborol-2-yl]butyl}amino)-2-oxoethyl]benzamide as a white solid, which was used in the next step without purification.

[0321] Step 3: N,N',N''-{boroxine-2,4,6-tolyltris[[(1R)-3-methylbutane-1,1-diyl]imino(2-oxoethane-2,1-diyl)]}tris(2,5-dichlorobenzamide) 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-benzodioxaborol-2-yl]butyl}amino)-2-oxoethyl]benzamide (12.2 g, 24.6 mmol) in methanol / hexane (1:1) (250 mL) was added 1 N HCl (30 mL, 30 mmol) and (2-methylpropyl)boronic acid (6.5 g, 64 mmol). The reaction mixture was allowed to stir overnight. The phases were separated, and the methanol layer was washed twice with additional heptane (2 × 55 mL). The resulting organic layer was concentrated to approximately 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 twice with DCM (2 × 25 mL), and acidified with 1 M HCl (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 × 10 mL). The combined DCM extracts were dried over MgSO (25 g) and evaporated to a thick oil. The product was precipitated with heptane (50 mL) and collected by filtration to give N,N',N''-{boroxine-2,4,6-tolyltris[[(1R)-3-methylbutane-1,1-diyl]imino(2-oxoethane-2,1-diyl)]}tris(2,5-dichlorobenzamide) (6.6 g, 74%) as a white solid. 1 H NMR (300MHz, DMSO-d6, δ): 8.93(t,J=6.0Hz,1H), 8.68(bs,1H), 7.63(m,1H), 7.52(m,2H), 4.0 0(d,J=6.0Hz,2H), 2.62(m,1H), 1.59(m,1H), 1.33(m,1H), 1.24(m,1H), 0.81(d,J=5.9Hz,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 52Calculated for B3Cl6N6O9 1027.2; Found 1027.3; [M+Na] C 42 H 51 Calculated for B3Cl6N6NaO9: 1049.2; Found: 1049.5.

[0322] Step 4: 4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2 ,5-Dichlorobenzamido)acetamido)-3-methylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid (I-1) Form 1: To a solution of citric acid (2.75 g, 14.3 mmol) in EtOAc (85 mL) having an internal temperature of about 74° C. was added N,N′,N″-{boroxine-2,4,6-tolyltris[[(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 until the internal temperature was about 25° C., and the mixture was stirred overnight. The resulting precipitate was collected by filtration to give 2,2′-{2-[(1R)-1-({[(2,5-dichlorobenzoyl)amino]acetyl}amino)-3-methylbutyl]-5-oxo-1,3,2-dioxaborolane-4,4-diyl}diacetic acid Form 1 (6.65 g, 88%) as a crystalline solid. 1 H NMR (500MHz, DMSO-d6, δ110℃): 10.08(s,1H), 8.69(s,1H), 7.61(s,1H), 7.5 2(d,J=1.3Hz,2H), 4.26(d,J=5.5Hz,2H), 2.70(q,J=14.5Hz,4H), 2.70(bs,1 H), 1.72(sept,J=6.5Hz,1H), 1.42(ddd,J=5.2Hz,J=8.6Hz,J=13.9Hz,1H), 1.28(ddd,J=5.3,J=9.4Hz,J=14.3Hz,1H),0.91(dd,J=3.3Hz,J=6.6Hz,6H). MS (m / z) in CH3CN: [M+Na] C 20 H 23 Calculated for BCl2N2NaO9 539.1; found 539.1.

[0323] XRPD data for I-1 Form 1 is shown in FIG.

[0324] [Table 1] Differential scanning calorimetry (DSC) data for I-1 Form 1 is shown in Figure 2. The profile is characterized by an endothermic transition with an onset temperature of 191.8°C and a melting point of 198.8°C. A second endothermic transition, corresponding to decomposition, has an onset temperature of 225°C. These temperatures have an error of ±5°C.

[0325] Thermogravimetric analysis (TGA) data for I-1 Form 1 is shown in Figure 2. The profile graphs the weight loss rate of the sample as a function of temperature, where the temperature change rate is approximately 10°C / min. The weight loss indicates a loss of approximately 0.72% of the sample's weight as the temperature changes from 50°C to 200°C. These temperatures have an error of ±5°C.

[0326] Form 2: To a solution of citric acid (10.1 g, 52.6 mmol) in EtOAc (300 mL) having an internal temperature of about 74° C. was added a solution of N,N′,N″-{boroxine-2,4,6-tolyltris[[(1R)-3-methylbutane-1,1-diyl]imino(2-oxoethane-2,1-diyl)]}tris(2,5-dichlorobenzamide) (20.0 g, 19.5 mmol) in EtOAc (60 mL). The solution was cooled slowly (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 cooled slowly (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 give 4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-methylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid Form 2 (26.7 g, 98%) as a crystalline solid. 1H NMR (500MHz, DMSO-d6, δ110℃): 10.08(s,1H), 8.69(s,1H), 7.61(s,1H), 7.5 2(d,J=1.3Hz,2H), 4.26(d,J=5.5Hz,2H), 2.70(q,J=14.5Hz,4H), 2.70(bs,1 H), 1.72(sept,J=6.5Hz,1H), 1.42(ddd,J=5.2Hz,J=8.6Hz,J=13.9Hz,1H), 1.28(ddd,J=5.3,J=9.4Hz,J=14.3Hz,1H),0.91(dd,J=3.3Hz,J=6.6Hz,6H). 13 C 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) in CH3CN: [M+Na] C 20 H 23 Calculated for BCl2N2NaO9 539.1; found 539.1.

[0327] 4-(R,S)-(Carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-methylbutyl)-6-oxo-1,3,2-dioxaborinane-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''-{boroxine-2,4,6-tolyltris[[(1R)-3-methylbutane-1,1-diyl]imino(2-oxoethane-2,1-diyl)]}tris(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 EtOAc addition was complete, the temperature was allowed to decrease to 20° C. over 5 hours. The resulting suspension was filtered to collect 4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-methylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid Form 2 (40 g, 70%) as a crystalline solid.

[0328] 4-(R,S)-(Carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-methylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid Form 2 was also prepared in the same general manner using the conditions described in Table 2.

[0329] [Table 2] 4-(R,S)-(Carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-methylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid Form 2 was also prepared by dissolving in acetone and then adding EtOAc as an anti-solvent.

[0330] XRPD data for Form I-1 is shown in Figure 3 and Table 3.

[0331] [Table 3] Differential scanning calorimetry (DSC) data for I-1 Form 2 is shown in Figure 4. The profile is characterized by an endothermic transition with an onset temperature of 206.5°C and a melting point of 219.9°C. A second endothermic transition, corresponding to decomposition, has an onset temperature of 225°C. These temperatures have an error of ±5°C.

[0332] Thermogravimetric analysis (TGA) data for I-1 Form 2 is shown in Figure 4. The profile graphs the weight loss rate of the sample as a function of temperature, where the temperature change rate is approximately 10°C / min. The weight loss indicates a loss of approximately 1.1% of the sample's weight as the temperature changes from 50°C to 200°C. These temperatures have an error of ±5°C.

[0333] Example 1A: Alternate synthesis of 4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-methylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid (I-1) form 2 Equipped with a mechanical stirrer, dropping funnel, thermometer, and heating / cooling control unit (under nitrogen). A 50 L glass reactor was charged with 1.2 micron filtered EtOAc (18.9 kg) and anhydrous citric acid (0.561 kg, 2.9 mol). The mixture was heated to 71°C to obtain a solution. N,N',N''-{boroxine-2,4,6-tolyltris[[(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 under stirring (193 rpm) while maintaining the temperature at 73°C to 75°C. The stirring was reduced to 96 rpm, and the mixture was cooled as follows: (1) The mixture was maintained at 73-75°C for 25 minutes; (2) The mixture was gradually cooled to 40°C at a rate of approximately 5°C / 30 minutes; (3) The mixture was allowed to cool to ambient temperature overnight with stirring. 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 yield 1.458 kg (92%) of the title compound. 1 H NMR(400MHz,DMSO-d6,δ):12.13(s,2H), 10.69(s,1H), 9.11(t,J=5.6Hz,1H), 7.66(t,J=1.2Hz,1H), 7.56 (d,J=1.2Hz,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=6.4Hz,6H).

[0334] XRPD data for compound (I-1) Form 2 is shown in FIG.

[0335] [Table 6] Differential scanning calorimetry (DSC) data for Compound (I-1) Form 2 is shown in Figure 8. The profile is characterized by two endothermic transitions, a first with a melting point of about 231.3°C and a second with a melting point of about 239.9°C. These temperatures are There is an error of ±5℃.

[0336] 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 (I-2) To a solution of glycolic acid (0.041 g, 0.54 mmol) in EtOAc (2.0 mL) having an internal temperature of about 60° C. was added a solution of N,N′,N″-{boroxine-2,4,6-tolyltris[[(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 until the internal temperature was about 25° C., and the solvent was removed by evaporation to give 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) in CHCN: [M+EtN+H] C 22 H 35 Calculated for BCl2N3O5 502.2; found 502.0. MS (m / z) in CH3CN: [MH]C 16 H 18 Calculated for BCl2N2O5 399.1; found 399.0.

[0337] 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 (I-3) To a solution of L-malic acid (0.0958 g, 0.714 mmol) in EtOAc (2.0 mL) having an internal temperature of about 60° C. was added a solution of N,N′,N″-{boroxine-2,4,6-tolyltris[[(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 to an internal temperature of approximately 25° C. and the solvent was removed by evaporation to give {(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) in CH3CN: [M+Et3N+H] 24 H 37 Calculated for BCl2N3O7 560.1; Found 560.1. MS (m / z) in CH3CN: [MH]C 18 H 20 Calculated for BCl2N2O7 457.1; found 457.1.

[0338] 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 (I-4) To a solution of (S)-hexahydromandelic acid (0.0881 g, 0.557 mmol) in EtOAc (2.0 mL) having an internal temperature of about 60° C. was added a solution of N,N′,N″-{boroxine-2,4,6-tolyltris[[(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 to an internal temperature of approximately 25° C. and the solvent was removed by evaporation to give 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] 28 H45 Calculated for BCl2N3O5 584.3; found 584.1. MS (m / z) in CH3CN: [MH]C 22 H 28 Calculated value for BCl2N2O5: 481.1 ;Actual value: 481.1.

[0339] 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 (I-5) To a solution of 2-hydroxyisobutyric acid (0.0567 g, 0.545 mmol) in EtOAc (2.0 mL) having an internal temperature of about 60° C. was added a solution of N,N′,N″-{boroxine-2,4,6-tolyltris[[(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 to an internal temperature of approximately 25° C. and the solvent was removed by evaporation to give 2,5-dichloro-N-(2-{[(1R)-1-(4,4-dimethyl-5-oxo-1,3,2-dioxaborolan-2-yl)-3-methylbutyl]amino}-2-oxoethyl)benzamide (0.225 g, 96%) as a white solid. MS (m / z) in CH3CN: [M+Et3N+H] 24 H 39 Calculated for BCl2N3O5 530.2; Found 530.0. MS (m / z) in CH3CN: [MH]C 18 H 22 Calculated for BCl2N2O5 427.1; found 427.0.

[0340] 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) To a solution of (R)-mandelic acid (0.168 g, 1.10 mmol) in EtOAc (2.0 mL) having an internal temperature of about 60° C. was added a solution of N,N′,N″-{boroxine-2,4,6-tolyltris[[(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 until the internal temperature was approximately 25° C. and the resulting precipitate was collected by filtration to give 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. 1 H NMR(300MHz,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=6.0Hz,2H), 2.8-2.76(m,1H), 1.71-1.62(m,1H), 1.50-1.28(m,2H), 0.89(m,6H). MS in CH3CN (m / z):[M+Et3N+H] C 28 H 39 Calculated for BCl2N3O5 578.2; found 578.1. MS (m / z) in CH3CN: [MH]C 22 H 22 Calculated for BCl2N2O5 475.1; found 475.1.

[0341] 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) To a solution of L-lactic acid (0.675 g, 7.34 mmol) in EtOAc (3.0 mL) having an internal temperature of approximately 70° C. was added a solution of N,N′,N″-{boroxine-2,4,6-tolyltris[[(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 until the internal temperature was approximately 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 approximately 70° C., at which point a homogeneous solution was obtained. The solution was cooled at a rate of 0.17° C. / min until the internal temperature was approximately 30° C. The mixture was then allowed to cool to an internal temperature of approximately 0° C. The resulting precipitate was collected by filtration to give 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 CHCN: [M+EtN+H] 23 H 37 Calculated for BCl2N3O5 515.9; Found 516.0. MS (m / z) in CH3CN: [MH]C 17 H 20 Calculated for BCl2N2O5 413.1; found 413.0.

[0342] The XRPD data for I-7 is shown in FIG.

[0343] [Table 4] 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) To a solution of (S)-3-hydroxybutyric acid (0.0598 g, 0.566 mmol) in EtOAc (2.0 mL) having an internal temperature of about 60° C. was added a solution of N,N′,N″-{boroxine-2,4,6-tolyltris[[(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 to an internal temperature of approximately 25° C. and the solvent was removed by evaporation to give 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(300MHz,DMSO-d6,δ):10.45(s,1H), 9.11(t,J=6.0Hz,1H), 7.65(m,1H), 7.55(m,2H), 4.21(d,J=6.0Hz,2H), 3.98-3.90(m,1H), 2.5 1(m,1H), 2.33(dd,J1=19.2Hz,J=2.7Hz,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 in CH3CN (m / z):[M+ Et3N+H] C 24 Calculated for H39BCl2N3O5 530.2; found 530.0. MS (m / z) in CH3CN: [MH]C 18 H 22 Calculated for BCl2N2O5 427.1; found 427.1.

[0344] Example 9: Synthesis of 2,5-dichloro-N-(2-{[(1R)-1-(4,4-dimethyl-6-oxo-1,3,2-dioxaborinan-2-yl)-3-methylbutyl]amino}-2-oxoethyl)benzamide (I-9) To a solution of β-hydroxyisovaleric acid (0.0841 g, 0.712 mmol) in EtOAc (2.0 mL) having an internal temperature of about 60° C. was added a solution of N,N′,N″-{boroxine-2,4,6-tolyltris[[(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 to an internal temperature of approximately 25° C. and the solvent was removed by evaporation to give 2,5-dichloro-N-(2-{[(1R)-1-(4,4-dimethyl-6-oxo-1,3,2-dioxaborinan-2-yl)-3-methylbutyl]amino}-2-oxoethyl)benzamide (0.296 g, 95%) as a white solid. MS (m / z) in CH3CN: [M+ Et3N+H] 25 H 41 Calculated for BCl2N3O5 544.3; found 544.0. MS (m / z) in CH3CN: [MH]C 19 H 24 Calculated for BCl2N2O5 441.1; found 441.0.

[0345] 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 (I-10) To a solution of (S)-2-hydroxy-3,3-dimethylbutyric acid (0.0712 g, 0.553 mmol) in EtOAc (2.0 mL) having an internal temperature of about 60° C. was added a solution of N,N′,N″-{boroxine-2,4,6-tolyltris[[(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 to an internal temperature of approximately 25° C. and the solvent was removed by evaporation to give 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 (0.245 g, 97%) as a white solid. MS (m / z) in CH3CN ):[M+ Et3N+H] C 26 H 43 Calculated for BCl2N3O5 558.3; found 558.0. MS (m / z) in CH3CN: [MH]C 20 H 26 Calculated for BCl2N2O5 455.1; found 455.0.

[0346] Example 11: Synthesis of 2,5-dichloro-N-[2-({(1R)-1-[(4S)-4-isopropyl-5-oxo-1,3,2-dioxaborolan-2-yl]-3-methylbutyl}amino)-2-oxoethyl]benzamide (I-11) To a solution of (S)-2-hydroxy-3-methylbutyric acid (0.0659 g, 0.558 mmol) in EtOAc (2.0 mL) having an internal temperature of about 60° C. was added a solution of N,N′,N″-{boroxine-2,4,6-tolyltris[[(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 to an internal temperature of approximately 25° C. and the solvent was removed by evaporation to give 2,5-dichloro-N-[2-({(1R)-1-[(4S)-4-isopropyl-5-oxo-1,3,2-dioxaborolan-2-yl]-3-methylbutyl}amino)-2-oxoethyl]benzamide (0.246 g, 99%) as a white solid. MS (m / z) in CH3CN: [M+Na] C 19 H 25 Calculated for BClNNaO 465.1; found 465.1. MS (m / z) in CHCN: [MH]C 19 H 24 Calculated for BCl2N2O5 441.1; found 441.0.

[0347] 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 (I-12) To a solution of 2-hydroxyisocaproic acid (0.0752 g, 0.569 mmol) in EtOAc (2.0 mL) having an internal temperature of about 60° C. was added a solution of N,N′,N″-{boroxine-2,4,6-tolyltris[[(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 to an internal temperature of approximately 25° C. and the solvent was removed by evaporation to give 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) in CH3CN: [M+Na] C 20 H 27 Calculated for BClNNaO 479.1; found 479.1. MS (m / z) in CHCN: [MH] C 20 H 26 Calculated for BCl2N2O5 455.1; found 455.1.

[0348] Example 13: Synthesis of 2,5-dichloro-N-(2-{[(1R)-3-methyl-1-(4-oxo-4H-1,3,2-benzodioxaborinin-2-yl)butyl]amino}-2-oxoethyl)benzamide (I-13) To a solution of salicylic acid (0.0758 g, 0.549 mmol) in EtOAc (2.0 mL) having an internal temperature of about 60° C. was added a solution of N,N′,N″-{boroxine-2,4,6-tolyltris[[(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 to an internal temperature of approximately 25° C. and the resulting precipitate was collected by filtration to give 2,5-dichloro-N-(2-{[(1R)-3-methyl-1-(4-oxo-4H-1,3,2-benzodioxaborinin-2-yl)butyl]amino}-2-oxoethyl)benzamide (0.198 g, 78%) as a white solid. S(m / z):[M+Na] C 21 H21 Calculated for BClNNaO 485.1; found 485.1. MS (m / z) in CHCN: [MH]C 21 H 20 Calculated for BCl2N2O5 461.1; found 461.0.

[0349] XRPD data for I-13 is shown in FIG.

[0350] [Table 5] Example 14: 2,5-Dichloro-N-(2-{[(1R)-3-methyl-1-(5-oxo-4,4-diphenyl-1,3,2-dioxaborolan-2-yl)butyl]amino Synthesis of}-2-oxoethyl)benzamide (I-14) To a solution of benzilic acid (0.126 g, 0.552 mmol) in EtOAc (2.0 mL) having an internal temperature of about 60° C. was added a solution of N,N′,N″-{boroxine-2,4,6-tolyltris[[(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 to an internal temperature of approximately 25° C. and the solvent was removed by evaporation to give 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) in CH3CN: [M+Na] C 28 H 27 Calculated for BClNNaO 5 575.1; found 575.2. MS (m / z) in CHCN: [MH] C 28 H 26 Calculated for BCl2N2O5 551.1; found 551.1.

[0351] Example 15: Synthesis of 2,2'-{2-[(1R)-3-methyl-1-({(2S)-3-phenyl-2-[(pyrazin-2-ylcarbonyl)amino]propanoyl}amino)butyl]-5-oxo-1,3,2-dioxaborolane-4,4-diyl}diacetic acid (I-15) To a solution of citric acid (0.257 g, 1.34 mmol) in EtOAc (7.4 mL) having an internal temperature of about 74° C. was added N,N′,N″-(boroxine-2,4,6-tolyltris{[(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 to an internal temperature of approximately 25° C. and evaporated to give 2,2′-{2-[(1R)-3-methyl-1-({(2S)-3-phenyl-2-[(pyrazin-2-ylcarbonyl)amino]propanoyl}amino)butyl]-5-oxo-1,3,2-dioxaborolane-4,4-diyl}diacetic acid (0.730 g, 99%) as a white solid. MS (m / z) in CH3CN: [M+Et3N+H] 31 H 45 Calculated for BN5O9: 642.3; Found: 642.2. MS (m / z) in CH3CN: [MH] C 25 H 28 Calculated for BN4O9 539.2; Found 539.2.

[0352] 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 (I-16) To a solution of (R)-mandelic acid (0.0738 g, 0.485 mmol) in EtOAc (2.0 mL) having an internal temperature of about 60° C. was added N,N′,N″-(boroxine-2,4,6-tolyltris{[(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 until the internal temperature was approximately 25° C. and the resulting precipitate was collected by filtration to give 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) in CHCN: [M+Na] C 27 H29 Calculated for BN4NaO5 523.2; Found 523.2. MS (m / z) in CH3CN: [MH]C 27 H 28 Calculated for BN4O5 499.2; Found 499.2.

[0353] Example 17: N-[(1S)-1-benzyl-2-({(1R)-3-methyl-1-[(5R)-4-oxo-5-phenyl-1,3,2-dioxaborolan-2-yl]butyl] Synthesis of ((2-(2-methyl-2-oxoethyl)amino)pyrazine-2-carboxamide (I-17) To a solution of (S)-3-hydroxybutyric acid (0.0509 g, 0.489 mmol) in EtOAc (2.0 mL) having an internal temperature of about 60° C. was added N,N′,N″-(boroxine-2,4,6-tolyltris{[(1R)-3-methylbutane-1,1-diyl]imino[(2S)-1-oxo-3-phenylpropane-1,2-diyl]})tripyrazine-2-carboxamide (0.179 g, 0.163 mmol) as a solid. The solution was allowed to cool to an internal temperature of approximately 25° C. and the solvent was removed by evaporation to give N-[(1S)-1-benzyl-2-({(1R)-3-methyl-1-[(4S)-4-methyl-6-oxo-1,3,2-dioxaborinan-2-yl]butyl}amino)-2-oxoethyl]pyrazine-2-carboxamide (0.213 g, 96%) as a white solid. MS (m / z) in CHCN: [M+Na] C 23 H 29 Calculated for BN4NaO5 475.2; Found 475.2. MS (m / z) in CH3CN: [MH]C 23 H 28 Calculated for BN4O5: 451.2; Found: 451.1.

[0354] Example 18: Preparation of a formulation of 4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-methylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid (I-1) for parenteral or oral administration Formulation A: A bottle was filled with 90 mL of water, and citric acid monohydrate (0.08 g) and sodium citrate dihydrate (1.5 g) were added and stirred until dissolved. 4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-methylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid (I-1) Form 2 (0.142 g) was added, and the mixture was stirred until a solution was obtained. Sodium chloride (0.45 g) was added to the solution, and the pH was adjusted to 5.45 with 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 obtain Formulation A, which was stored at −20° C.

[0355] Formulation B was prepared as Formulation A, except that the pH was adjusted to pH 6.2 with 2N NaOH.

[0356] Formulation C: A bottle was filled with 90 mL of water, and citric acid monohydrate (0.08 g), sodium citrate dihydrate (1.5 g), and propylene glycol (1.0 g) were added and stirred until dissolved. 4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-methylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid (I-1) Form 2 (0.142 g) was added to the solution, and the mixture was stirred until a solution was obtained. The pH was adjusted to 6.2 with 2 N NaOH, and the final volume of the resulting solution was adjusted to 100 mL with water and filtered through a 0.2 μm PES filter to obtain Formulation C, which was stored at −20° C.

[0357] Example 19: In situ preparation of formulations of 4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-methylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid (I-1) for parenteral or oral administration Preservative formulation vehicle: A bottle was filled with approximately 160 mL of water, and citric acid monohydrate (0.714 g) and sodium citrate dihydrate (2.24 g) were 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 resulting solution was stirred until the solution was dissolved. The final pH was pH 5.14. The final volume of the resulting solution was adjusted to 200 g (assuming a concentration of 1 g / mL) with water, filtered through a 0.2 μm PES membrane filter unit, and stored at a temperature of about 2°C to about 8°C.

[0358] Formulation preservative (1 mg / mL): To a bottle was added 0.105 grams (approximately 95.4% purity) of N,N',N''-{boroxine-2,4,6-tolyltris[[(1R)-3-methylbutane-1,1-diyl]imino(2-oxoethane-2,1-diyl)]}tris(2,5-dichlorobenzamide). To this was added approximately 90 g of preservative formulation vehicle, and the resulting mixture was stirred for 48 hours while protected from light. The final pH was pH 5.12. The final volume of the resulting solution was adjusted to 100 g (assuming a concentration of 1 g / mL) with the preservative formulation vehicle, filtered through a 0.2 μm PES membrane filter unit, and stored at a temperature of about 2°C to about 8°C while protected from light.

[0359] Formulation D: The formulation preservative was diluted to a concentration of 0.05 mg / mL to 0.1 mg / mL with the preservative formulation vehicle prior to use.

[0360] Formulation E: The formulation preservative was diluted with 0.9% sodium chloride solution to a concentration of 0.05 mg / mL to 0.1 mg / mL before use.

[0361] Example 20: 20S proteasome assay In a 384-well black microtiter plate, 1 μL of test compound dissolved in DMSO was added to 25 μL of assay buffer containing human PA28 activator (Boston Biochem, 12 nM final) along with Ac-WLA-AMC (β5-selective substrate) (15 μM final) at 37°C, followed by 25 μL of assay buffer containing human 20S proteasome (Boston Biochem, 0.25 nM final) at 37°C. The assay buffer consisted of 20 mM HEPES, 0.5 mM EDTA, and 0.01% BSA, pH 7.4. Reactions were run on a BMG Galaxy plate reader (37°C, excitation 380 nm, emission 460 nm, amplification 20). 0% Percent inhibition is calculated relative to controls of no inhibition (DMSO) and 100% inhibition (10 μM bortezomib).

[0362] Example 21: Antiproliferative assay 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) are seeded into wells of a 96-well cell culture plate and incubated overnight at 37°C. Test compounds are added to the wells, and the plate is incubated at 37°C for 96 hours. MTT or WST reagent (10 μL, Roche) is added to each well as described by the manufacturer and incubated at 37°C for 4 hours. The MTT metabolic stain is solubilized overnight according to the manufacturer's instructions (Roche). The optical density for each well is read using a spectrophotometer (Molecular Devices) at 595 nm (initial) and 690 nm (reference) for MTT, and 450 nm for WST. The reference optical density value for MTT is subtracted from the initial wavelength value. Percent inhibition is calculated using the values ​​from the DMSO control set relative to 100%.

[0363] Example 22: In vivo tumor efficacy model Freshly dissociated HCT-116 (2-5 × 10) cells in 100 μL of RPMI-1640 medium (Sigma-Aldrich) were cultured. 6 ) or other tumor cells were injected into the subcutaneous space in the right dorsal flank of female CD-1 nude mice (5-8 weeks old, Charles River) using a 1 mL 26 3 / 8 gauge needle (Becton Dickinson Ref#309625). Alternatively, some 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 dorsal flank of anesthetized (3-5% isoflorane / oxygen mixture) CB-17 / SCID mice (5-8 weeks old, Charles River) via a 13-gauge trocar (Popper & Sons 7927). Beginning on day 7 post-inoculation, tumors are measured twice weekly using calipers. Tumor volume is calculated using standard procedures as (0.5 x (length x width)). 2 The tumor was about 200 mm 3 When the tumor volume reaches 1000 mm, mice are randomized into treatment groups and begin receiving drug therapy. Dosing and schedule are determined for each experiment based on previous results from pharmacokinetic / pharmacodynamic and maximum tolerated dose studies. Control groups may receive vehicle without any drug. Generally, test compounds (100-200 μL) are administered via intravenous (27-gauge needle), oral (20-gauge gavage needle), or subcutaneous (27-gauge needle) routes at various doses and schedules. Tumor size and body weight are measured twice weekly, and the study is completed when control tumors reach approximately 2000 mm. 3 It is terminated when it reaches

[0364] Example 23: Synthesis of N-((S)-1-((R)-3-methyl-1-(4-oxo-4H-benzo[d][1,3,2]dioxaborinin-2-yl)butylamino)-1-oxo-3-phenylpropan-2-yl)pyrazine-2-carboxamide (I-19) A mixture of N,N',N''-(boroxine-2,4,6-tolyltris{[(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 until the internal temperature was approximately 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]dioxaborinin-2-yl)butylamino)-1-oxo-3-phenylpropan-2-yl)pyrazine-2-carboxamide (0.249 g, 75%). MS (m / z) in CHCN: [M+H] C 26 H 28 Calculated value for BN4O5: 487.2153; Found: 487.3.

[0365] Example 24: Synthesis of 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 (I-20) A mixture of N,N',N''-(boroxine-2,4,6-tolyltris{[(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 until the internal temperature was approximately 25°C. A white solid precipitated, 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-1-((S)-3-phenyl-2-(pyrazine-2-carboxamido)propanamido)butyl)-5-oxo-1,3,2-dioxaborolan-4-yl)acetic acid (0.625 g, 95%). MS (m / z) in CHCN: [M+H] C 23 H 28 Calculated value for BN4O7: 483.2051; found value: 483.2.

[0366] Example 25: Synthesis of 2-((R)-2-((R)-3-methyl-1-((S)-3-phenyl-2-(pyrazine-2-carboxamido)propanamido)butyl)-5-oxo-1,3,2-dioxaborolan-4-yl)acetic acid (I-21) N,N',N''-(boroxine-2,4,6-tolyltris{[(1R)-3-methyl A mixture of [(R)-2-((R)-3-methyl-1-((S)-3-phenyl-2-(pyrazine-2-carboxamido)propanamido)butyl]-5-oxo-1,3,2-dioxaborolan-4-yl)acetic acid (0.410 g, 100%) was mixed in acetone (3 mL). The mixture was heated to form a solution. The solution was allowed to cool until the internal temperature reached approximately 25°C. A white solid precipitated, 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-1-((S)-3-phenyl-2-(pyrazine-2-carboxamido)propanamido)butyl)-5-oxo-1,3,2-dioxaborolan-4-yl)acetic acid (0.410 g, 100%). [M+H] C 23 H 28 Calculated value for BN4O7: 483.2051; found value: 483.2.

[0367] Example 26: Synthesis of (R)-2-hydroxy-2-((R)-2-((R)-3-methyl-1-((S)-3-phenyl-2-(pyrazine-2-carboxamido)propanamido)butyl)-5-oxo-1,3,2-dioxaborolan-4-yl)acetic acid (I-22) A mixture of N,N',N''-(boroxine-2,4,6-tolyltris{[(1R)-3-methylbutane-1,1-diyl]imino[(2S)-1-oxo-3-phenylpropane-1,2-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 until the internal temperature reached approximately 25°C. Butane (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, and solid (R)-2-hydroxy-2-((R)-2-((R)-3-methyl-1-((S)-3-phenyl-2-(pyrazine-2-carboxamido)propanamido)butyl)-5-oxo-1,3,2-dioxaborolan-4-yl)acetic acid (0.388 g) was collected, which also contained dimeric species. MS (m / z) in CH3CN: [M+H] C 23 H 28 Calculated for BN4O8: 499.2000; Found: 499.2.

[0368] Example 27: Synthesis of (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 (I-23) A mixture of N,N',N''-(boroxine-2,4,6-tolyltris{[(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 until the internal temperature was approximately 25°C. Heptane (8 mL) was added. The mixture was evaporated to give (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 species. MS (m / z) in CHCN: [M+H] C 23 H 28Calculated for BN4O8: 499.2000; Found: 499.2.

[0369] Example 28: Pharmaceutical Composition 1 The composition of the capsules is shown in Table 7 below.

[0370] [Table 7] Example 29: Pharmaceutical Composition 2 The composition of the capsules is shown in Table 8 below.

[0371] [Table 8] Example 30: Pharmaceutical Composition 3 The composition of the capsules is shown in Table 9 below.

[0372] [Table 9] Example 31: Pharmaceutical Composition 4 The composition of the capsules is shown in Table 10 below.

[0373] [Table 10] Example 32: Pharmaceutical Composition 5 The composition of the capsules is shown in Table 11 below.

[0374] [Table 11] Example 33: Pharmaceutical Composition 6 The composition of the capsules is shown in Table 12 below.

[0375] [Table 12] Example 34: Pharmaceutical Composition 7 The composition of the capsules is shown in Table 13 below.

[0376] [Table 13] Example 35: Pharmaceutical Composition 8 The compositions are shown in Table 14 below.

[0377] [Table 14] The batches were prepared according to the following process. 1) Microcrystalline cellulose, NF (Emcocel® XLM90, low moisture) (item number #2) was sieved through a 40 micrometer mesh screen. 2) The sieved material from step 1) was added to a PK mixer and blended for 2 minutes. 3) Compound of formula (I-1) Form 2, sieved through a 60 micrometer mesh sieve, was weighed out (item number #1). 4) The compound of formula (I-1) Form 2 from step 3) and microcrystalline cellulose, NF (Emcocel® XLM90, low moisture) (item no. #3) are mixed in a polyethylene terephthalate (PE) emulsion. The mixture was mixed in the bag, the polyethylene bag was shaken, and the contents of the polyethylene bag were then passed through the same 40 micrometer sieve as used in step 1). 5) The material from step 4) was added to the PK mixer and blended for 15 minutes. 6) Microcrystalline cellulose, NF (Emcocel® XLM90, low moisture) (item number #4) was sieved through the same 40 micrometer mesh sieve, transferred to the same polyethylene bag used in step 4), and shaken in the polyethylene bag. 7) The material from step 6) was added to the PK mixer, which still contained the material from step 5), and blended for 10 minutes. 8) Microcrystalline cellulose, NF (Emcocel® XLM90, low moisture) (item number #5) was sieved through the same 40 micrometer mesh sieve, transferred to the same polyethylene bag used in steps 4) and 6), and shaken in the polyethylene bag. 9) The material from step 8) was added to the PK mixer, which still contained the material from steps 5) and 7), and blended for 10 minutes. 10) Microcrystalline cellulose, NF (Emcocel® XLM90, low moisture) (item number #6) was sieved through the same 40 micrometer mesh sieve and transferred to the same polyethylene bag used in steps 4), 6), and 8) and shaken in the polyethylene bag. 11) The material from step 10) was added to the PK mixer, which still contained the material from steps 5) and 7), and 9) and blended for 10 minutes. 12) The material from the mixer was encapsulated into No. 4 milky white gelatin capsules using the In-Cap system. 13) The capsules were de-dusted and sorted by weight.

[0378] Example 36: Pharmaceutical Composition 9 The composition of the capsules is shown in Table 15 below.

[0379] [Table 15] Example 37: Pharmaceutical Composition 10 The composition of the capsules is shown in Table 16 below.

[0380] [Table 16] Example 38: Pharmaceutical Composition 11 The composition of the capsules is shown in Table 17 below.

[0381] [Table 17] Example 39: Pharmaceutical Composition 12 The composition of the capsules is shown in Table 18 below.

[0382] [Table 18] Example 40: Pharmaceutical Composition 13 The composition of the capsules is shown in Table 19 below.

[0383] [Table 19] Example 41: Pharmaceutical Composition 14 The composition of the capsules is shown in Table 20 below.

[0384] [Table 20] Example 42: Pharmaceutical Composition 15 The composition of the capsules is shown in Table 21 below.

[0385] [Table 21] Example 43: Pharmaceutical Composition 16 The composition of the capsules is shown in Table 22 below.

[0386] [Table 22] The batches were prepared according to the following process. 1) Microcrystalline cellulose, NF (Emcocel® XLM90, low moisture) (item number #2) was sieved through a 40 micrometer mesh sieve. 2) The sieved material from step 1) was added to the PK mixer and blended for 2 minutes. 3) Compound of formula (I-1) Form 2 was sieved through a 60 micrometer mesh sieve and weighed (item no. #1). 4) The compound of formula (I-1) Form 2 from step 3) and microcrystalline cellulose, NF (Emcocel® XLM90, low moisture) (item number #3) were mixed in a polyethylene bag, the polyethylene bag was shaken, and the contents of the polyethylene bag were then passed through the same 40 micrometer sieve as used in step 1). 5) The materials from step 4) were added to a PK mixer and blended for 15 minutes. 6) Microcrystalline cellulose, NF (Emcocel® XLM90, low moisture) (item number #4) was sieved through the same 40 micrometer mesh sieve, transferred to the same polyethylene bag used in step 4), and shaken in the polyethylene bag. 7) Talc (item no. #7) and sodium citrate (item no. #8) were sieved through an identical 40 micrometer mesh sieve. 8) The materials from steps 6) and 7) were added to the PK mixer, which still contained the material from step 5), and blended for 10 minutes. 9) Microcrystalline cellulose, NF (Emcocel® XLM90, low moisture) (item number #5) was sieved through the same 40 micrometer mesh sieve, transferred to the same polyethylene bag used in steps 4) and 6), and shaken in the polyethylene bag. 10) The material from step 9) was added to the PK mixer, which still contained the material from steps 5) and 8), and blended for 10 minutes. 11) Microcrystalline cellulose, NF (Emcocel® XLM90, low moisture) (item number #6) was sieved through the same 40 micrometer mesh sieve, transferred to the same polyethylene bag used in steps 4), 6), and 9), and shaken in the polyethylene bag. 12) The material from step 11) was added to the PK mixer, which still contained the material from steps 5) and 8), and 10) and blended for 10 minutes. 13) Magnesium stearate (item #9) was sieved through the same 40 micrometer mesh sieve. 14) The material from step 13) was added to the PK mixer, which still contained the material from steps 5) and 8), 10), and 12) and blended for 5 minutes. 15) The material from the mixer was encapsulated into No. 4 milky white gelatin capsules using a Profill system. 16) The capsules were de-dusted and sorted by weight.

[0387] Example 44: Pharmaceutical Composition 17 The composition of the capsules is shown in Table 23 below.

[0388] [Table 23] The batches were prepared according to the following process. 1) Microcrystalline Cellulose, NF (Emcocel® XLM90, low moisture) (commercially available) Item #2) was sieved through a 40 micrometer mesh screen. 2) The sieved material from step 1) was added to a small PK mixer and blended for 2 minutes. 3) Compound of formula (I-1) Form 2 was sieved through a 60 micrometer mesh sieve and weighed (item no. #1). 4) The compound of formula (I-1) Form 2 from step 3) and microcrystalline cellulose, NF (Emcocel® XLM90, low moisture) (item no. #3) were mixed and then passed through the same 40 micrometer sieve as used in step 1). 5) The materials from step 4) were added to a small PK blender and blended for 30 minutes. 6) Microcrystalline cellulose, NF (Emcocel® XLM90, low moisture) (Item #4) and talc (Item #5) were sieved through an identical 40 micrometer mesh sieve. 7) The material from step 6) was added to the small PK blender, which still contained the material from step 5), and blended for 15 minutes. 8) Microcrystalline Cellulose, NF (Emcocel® XLM90, low moisture) (Item #6) was sieved through the same 40 micrometer mesh screen and transferred to a second large PK blender and blended for 2 minutes. 9) The contents of the small PK mixer from steps 5) and 7) were emptied into a polyethylene bag and then transferred to the large PK mixer from step 8). 10) Talc (Item #7) and microcrystalline cellulose, NF (Emcocel® XLM90, low moisture) (Item #8) were sieved through an identical 40 micrometer mesh sieve. 11) Half of the material from step 10) was added to the small PK mixer from steps 5) and 7), blended for 3 minutes, transferred to the same polyethylene bag used in step 9) and shaken in the polyethylene bag. 12) The material from step 11) was added to a large PK mixer, which still contained the material from steps 8) and 9). 13) The remaining half of the material from step 10) was added to the small PK mixer from steps 5), 7), and 11), blended for 3 minutes, transferred to the same polyethylene bag used in steps 9) and 11), and shaken in the polyethylene bag. 14) The material from step 13) was added to the large PK blender, which still contained the material from steps 8), 9), and 12) and blended for 10 minutes. 15) Microcrystalline cellulose, NF (Emcocel® XLM90, low moisture) (item no. #9) was sieved through the same 40 micrometer mesh sieve and transferred to the same polyethylene bag used in steps 9), 11), and 13) and shaken in the polyethylene bag. 16) The material from step 15) was added to the same large PK blender, which still contained the material from steps 8), 9), 12), and 14) and blended for 10 minutes. 17) Microcrystalline cellulose, NF (Emcocel® XLM90, low moisture) (item no. #10) was sieved through an identical 40 micrometer mesh sieve. 18) The material from step 17) was added to the same large PK blender, which still contained the material from steps 8), 9), 12), 14), and 16) and blended for 10 minutes. 19) Microcrystalline cellulose, NF (Emcocel® XLM90, low moisture) (item no. #11) was sieved through an identical 40 micrometer mesh sieve. 20) Add the material from step 19) to the same large PK mixer, which still contains the material from steps 8), 9), 12), 14), 16), and 18) and mix for 10 minutes. It was mixed. 21) Magnesium stearate (item no. #12) was sieved through the same 40 micrometer mesh sieve. 22) The material from step 21) was added to the same large PK blender, which still contained the material from steps 8), 9), 12), 14), 16), 18), and 20), and blended for 5 minutes. 23) The material from the mixer was encapsulated into No. 4 milky white gelatin capsules using the In-cap system. 24) The capsules were de-dusted and sorted by weight.

[0389] Example 45: Pharmaceutical Composition 18 The composition of the capsules is shown in Table 24 below.

[0390] [Table 24] The batches were prepared according to the following process. 1) Microcrystalline Cellulose, NF (Emcocel® XLM90, low moisture) (Item #2) is sieved through a 40 micrometer mesh sieve and added to a high shear mixer. 2) Sift the compound of formula (I-1) Form 2 through a 60 micrometer mesh sieve, weigh (Item #1) and add to the same high shear mixer from step 1). 3) Microcrystalline Cellulose, NF (Emcocel® XLM90, low moisture) (Item #3) is sieved through the same 40 micrometer mesh sieve and added to the same high shear mixer from steps 1) and 2). 4) The high shear mixer from steps 1), 2), and 3) is run for 4 minutes. 5) The material from the high shear mixer is encapsulated into No. 4 milky white gelatin capsules using an In-cap system. 6) Dust the capsules and classify them by weight.

[0391] Example 46: Lyophilized powder 1 In a clean container, a 40% tert-butyl alcohol / 60% water for injection solution was prepared by warming the required amount of tert-butyl alcohol to 35°C. The solution was cooled to 15-30°C. A portion of the required amount (60% of the total batch) of the tert-butyl alcohol / water solution was added to the pre-compound container. Approximately 40% of the solution was reserved for rinsing. Citric acid (30% of the batch amount) was added to the pre-compound container with stirring. The container was rinsed with the prepared tert-butyl alcohol / water solution, and the rinse was added to the pre-compound container. The mixture was stirred until the citric acid was completely dissolved. Sodium citrate (30% of the batch amount) was added to the pre-compound container with stirring. The container was rinsed with the prepared tert-butyl alcohol / water solution, and the rinse was added to the pre-compound container. The mixture was stirred until the sodium citrate was completely dissolved. N-(2-pyrazine)carbonyl-L-phenyl-L-leucineboronic acid (VIII-15) was added to the pre-compounding vessel with stirring. The vessel was rinsed with the prepared tert-butyl alcohol / water solution, and the rinse was added to the pre-compounding vessel. The mixture was stirred until the boronic acid was completely dissolved. The mixture of citric acid, sodium citrate, and boronic acid from the pre-compounding vessel was transferred to the main compounding bottle. The pre-compounding vessel was rinsed with water for injection, and the rinse was added to the main compounding bottle. Citric acid (70% of the batch amount) was added to the main compounding bottle with stirring. The vessel was rinsed with water, and the rinse was added to the main compounding bottle. The mixture was stirred until the citric acid was completely dissolved. Sodium citrate (70% of the batch amount) was added to the main compounding bottle with stirring. The vessel was rinsed with water, and the rinse was added to the pre-compounding vessel. The mixture was stirred until the sodium citrate was completely dissolved. Glycine was added to the main bottle, the residual glycine was rinsed with water, and the rinse was added to the main bottle. The mixture was stirred until the glycine was completely dissolved. Sufficient water was added to reduce the total alcohol content to 4.7% v / v. The mixture was filtered through a 0.22 μm filter. Aliquots of the filtered solution were placed in vials.The vials were sealed with lyophilization stoppers and placed on shelves in the lyophilizer chamber, maintained at 20°C. The shelves in the lyophilizer chamber were cooled to -45°C using an appropriate ramp rate and held at that temperature for 200 minutes. The shelves were warmed to -20°C using an appropriate ramp rate and held at that temperature for 480 minutes. The shelves were cooled again to -45°C using an appropriate ramp rate and held at that temperature. After 200 minutes, the lyophilization chamber was evacuated and the chamber pressure was adjusted to 150 micrometers with nitrogen. The chamber shelves were warmed to a maximum of -25°C using an appropriate ramp rate and held at that temperature for 3000 minutes. After each product thermocouple read above -25°C, the shelves were warmed to 27°C and held at that temperature for 600 minutes. At the end of the final drying phase, the chamber pressure was restored with nitrogen, and the vials were sealed and removed. The pre-lyophilized solution contained 5 mM citrate, 3% glycine, 4.7% tert-butyl alcohol (shown in Table 25 below).

[0392] [Table 25] Example 47: Lyophilized powder 2 Prepared as described in Example 46. The pre-lyophilized solution contained 52 mM citrate, 3% glycine, and 4.7% tert-butyl alcohol (shown in Table 26 below).

[0393] [Table 26] Example 48: Lyophilized powder 3 The formulation was prepared as described in Example 46, except for the lyophilization cycle was modified. Vials were sealed with lyophilization stoppers and placed on the shelves of the lyophilizer chamber, maintained at 20°C. The shelves of the lyophilization chamber were cooled to -45°C using an appropriate ramp rate and held at that temperature for 200 minutes. The shelves were warmed to -20°C using an appropriate ramp rate and held at that temperature for 480 minutes. The shelves were cooled again to -45°C using an appropriate ramp rate and held at that temperature. After 200 minutes, the lyophilization chamber was evacuated and the chamber pressure was adjusted to 150 micrometers with nitrogen. The shelves of the chamber were warmed to a maximum of -15°C using an appropriate ramp rate and held at that temperature for 2700 minutes. After each product thermocouple read above -15°C, the shelves were warmed to 37°C and held at that temperature for 300 minutes. At the end of the final drying phase, the chamber pressure was restored with nitrogen, and the vial was sealed and removed. The pre-lyophilized solution contained 52 mM citrate, 3% glycine, and 4.7% tert-butyl alcohol (shown in Table 27 below).

[0394] [Table 27] Example 49: Lyophilized powder 4 A clean bottle was filled with water for injection. Citric acid and sodium citrate were added and stirred until dissolved. To this solution, N-(2-pyrazine)carbonyl-L-phenyl-L-leucineboronic acid (VIII-15) was added and stirred until dissolved. Glycine was added to the bottle, residual glycine was rinsed with water, and the rinse was added to the main bottle. The mixture was stirred until the glycine was completely dissolved. Sufficient water was added to the batch volume. The mixture was filtered through a 0.22 μm filter. Aliquots of the filtered solution were placed into vials. The vials were sealed with lyophilization stoppers and placed on the shelves of the lyophilizer chamber, maintained at 20°C. The shelves of the lyophilizer chamber were cooled to -45°C using an appropriate ramp rate and held at that temperature for 200 minutes. The shelves were warmed to -20°C using an appropriate ramp rate and held at that temperature for 480 minutes. The shelves were cooled again to -45°C using an appropriate ramp rate and maintained at that temperature. After 200 minutes, the lyophilization chamber was evacuated and the chamber pressure was adjusted to 150 microns using nitrogen. The chamber shelves were warmed to a maximum of -25°C using an appropriate ramp rate and maintained at that temperature for 3000 minutes. After each product thermocouple read above -25°C, the shelves were warmed to 27°C and maintained at that temperature for 600 minutes. At the end of the final drying phase, the chamber pressure was restored using nitrogen, and the vials were sealed and removed. The pre-lyophilized solution contained 52 mM citrate and 3% glycine (shown in Table 28 below).

[0395] [Table 28] Example 50: Lyophilized Powder 5 The pre-lyophilized solution was prepared as described in Example 49. The pre-lyophilized solution contained 52 mM citrate and 3% glycine (shown in Table 29 below). In this example, the pH of the pre-lyophilized solution was adjusted to the final measured pH by adding 2N HCl.

[0396] [Table 29] Example 50: Lyophilized Powder 6 A clean bottle is filled with water for injection. Citric acid and sodium citrate are added and stirred until dissolved. 4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-methylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid (I-1) is added to this solution and stirred until dissolved. Glycine is added to the bottle, residual glycine is rinsed with water, and the rinse is added to the main bottle. The mixture is stirred until the glycine is completely dissolved. Sufficient water is added to the batch volume. The mixture is filtered through a 0.22 μm filter. Aliquots of the filtered solution are placed into sterile vials. The vials are sealed with lyophilization stoppers and placed on the shelves of the lyophilizer chamber, maintained at 20°C. The freeze-dryer chamber shelves are cooled to -45°C using an appropriate ramp rate and held at that temperature for 200 minutes. The shelves are warmed to -20°C using an appropriate ramp rate and maintained at that temperature for 480 minutes. The shelves are cooled again to -45°C using an appropriate ramp rate and maintained at that temperature. After 200 minutes, the freeze-drying chamber is evacuated and the chamber pressure is adjusted to 150 micrometers with nitrogen. The chamber shelves are warmed to a maximum of -25°C using an appropriate ramp rate and held at that temperature for 3000 minutes. After each product thermocouple reads -25°C or higher, the shelves are warmed to 27°C and maintained at that temperature for 600 minutes. At the end of the final drying phase, the chamber pressure is restored with nitrogen, and the vials are sealed and removed. The composition of the pre-lyophilized solution is 55 mM citrate and 3% glycine (shown in Table 30 below).

[0397] [Table 30] Example 51: Reconstitution of lyophilized powder The lyophilized powders (e.g., as prepared in Examples 46-50) are analyzed for cake structure, cake stability, residual solvent, and residual moisture using XRPD, DSC, gas chromatography, and Karl Fischer, respectively. The lyophilized powders are reconstituted with an appropriate amount of sterile water for injection or 0.9% sterile sodium chloride solution for injection. The reconstituted solutions are analyzed for ester purity and yield using HPLC and NMR.

[0398] Example 52: Preparation of a formulation of 4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-methylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid (I-1) Form 2 for parenteral or oral use A bottle was filled with water, and citric acid monohydrate and sodium citrate dihydrate were added and stirred until dissolved. To this solution, 4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-methylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid (I-1) Form 2 was added, and the mixture was stirred until a solution was obtained. To this solution, sodium chloride was added and stirred until dissolved. Sufficient water was added to the batch volume, and the solution was filtered through a 0.2 μm PES membrane. Aliquots of the filtered solution were placed in vials. The vials were stoppered and stored at −20° C. The batch and vial compositions are as set forth in Table 31 below.

[0399] [Table 31] Example 53: Analytical Test Method 1 Reversed-phase HPLC using a C8 column at 25°C with ultraviolet (UV) detection at 225 nm Mobile phase: The gradient system starts with 85% mobile phase A (0.01% trifluoroacetic acid in water) and 15% mobile phase B (0.01% trifluoroacetic acid in acetonitrile) and ends with 75% mobile phase B after 40 minutes.

[0400] Test samples are prepared by dissolving the capsule contents in a diluent consisting of 15:85 (v / v) acetonitrile:20 mM citrate buffer. Under these aqueous conditions, the compound of formula (I-1) completely hydrolyzes the citrate ester moiety to yield the compound of formula (VIII-1) in a 1:1 molecular ratio. The presence of the compound of formula (VIII-I) in the test sample is confirmed by comparing the retention time of the reference standard with that of the sample. The amount of the compound of formula (VIII-1) present in the sample is calculated from the area under the peak using the area under the peak of the reference standard based on a weight / weight comparison, including molecular weight conversion. The reference standard used is a known amount of compound of formula (I-1) of known purity, prepared under the same hydrolysis conditions as the test sample. The limit of quantification of the method is 0.05%, and the calculated limit of detection is 0.02%.

[0401] Example 54: Analytical Test Method 2 Normal-phase HPLC using isocratic elution with a mobile phase of 40 / 60 / 0.1 (v / v / v) THF / n-hexane / TFA on a cyano HPLC column for 8 min at 25°C with UV detection at 230 nm. Test samples were eluted in 40 / 60 (v / v) THF / n-hexane. The test sample is prepared by dissolving the contents of the capsule in HCl. Under these conditions, the compound of formula (I-1) is not hydrolyzed to the compound of formula (VIII-1). The amount of the compound of formula (VIII-1) present in the test sample is calculated from the area under the peak using the area under the peak of the reference standard based on a weight / weight comparison, including molecular weight conversion. The reference standard used is a known amount of the compound of formula (VIII-1) of known purity, which is prepared under the same hydrolysis conditions as the test sample. The quantitation limit for detection of the compound of formula (I-1) is 0.2%.

[0402] Both Analytical Test Method 1 and Analytical Test Method 2 are used to calculate the amount of compound of Formula (I-1) present in a test sample. Analytical Test Method 1 is used to calculate the amount, based on weight, of compound of Formula (VIII-1) present in a test sample containing compound of Formula (I-1). Analytical Test Method 2 is also used to calculate the amount of compound of Formula (VIII-1) present in a sample of compound of Formula (I-1) obtained without induced hydrolysis.

[0403] The amount of compound of Formula (VIII-1) obtained from Analytical Test Method 2 is subtracted from the amount of compound of Formula (VIII-1) obtained from Analytical Test Method 1 to obtain the measured amount of compound of Formula (VIII-1) produced by induced hydrolysis of compound of Formula (I-1) present in the test sample. The amount of compound of Formula (I-1) present in the test sample is obtained by calculating the molecular weight based on a 1:1 molecular ratio.

[0404] While the foregoing invention has been described in detail for purposes of clarity and understanding, these specific embodiments are intended to be illustrative and not restrictive. Those skilled in the art will recognize that various changes in form and detail may be made without departing from the true scope of the invention, which is to be defined by the appended claims rather than by the specific embodiments.

[0405] The patent and scientific literature referred to herein establishes knowledge that is available to those skilled in the art. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this invention belongs. Issued patents, patent applications, and references cited herein are incorporated by reference herein to the same extent as if each were specifically and individually indicated to be incorporated by reference. In the case of conflict, the present disclosure, including definitions, will control.

Claims

[Claim 1] Biological activity as described in the specification.

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