U.S. patent application number 16/788824 was filed with the patent office on 2020-06-11 for boronate ester compounds and pharmaceutical compositions thereof.
The applicant listed for this patent is Millennium Pharmaceuticals, Inc.. Invention is credited to Eric L. Elliott, Abu J. Ferdous, Michael J. Kaufman, Sonja A. Komar-Lay, Debra L. Mazaik, Quentin J. Mccubbin, Phuong M. Nguyen, Vaithianathan Palaniappan, Raymond D. Skwierczynski, Nobel T. Truong, Csanad M. Varga, Peter N. Zawaneh.
Application Number | 20200181176 16/788824 |
Document ID | / |
Family ID | 41061278 |
Filed Date | 2020-06-11 |
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United States Patent
Application |
20200181176 |
Kind Code |
A1 |
Elliott; Eric L. ; et
al. |
June 11, 2020 |
BORONATE ESTER COMPOUNDS AND PHARMACEUTICAL COMPOSITIONS
THEREOF
Abstract
The present invention provides novel compounds useful as
proteasome inhibitors. The invention also provides pharmaceutical
compositions comprising the compounds of the invention and methods
of using the compositions in the treatment of various diseases.
Inventors: |
Elliott; Eric L.; (Watetown,
MA) ; Ferdous; Abu J.; (Nottingham, MD) ;
Kaufman; Michael J.; (Lexington, MA) ; Komar-Lay;
Sonja A.; (Belmont, MA) ; Mazaik; Debra L.;
(Holliston, MA) ; Mccubbin; Quentin J.; (Belmont,
MA) ; Nguyen; Phuong M.; (Cambridge, MA) ;
Palaniappan; Vaithianathan; (Andover, MA) ;
Skwierczynski; Raymond D.; (Andover, MA) ; Truong;
Nobel T.; (Milford, MA) ; Varga; Csanad M.;
(Newton, MA) ; Zawaneh; Peter N.; (Brookline,
MA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Millennium Pharmaceuticals, Inc. |
Cambridge |
MA |
US |
|
|
Family ID: |
41061278 |
Appl. No.: |
16/788824 |
Filed: |
February 12, 2020 |
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16788824 |
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16406159 |
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15928139 |
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16163694 |
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15701791 |
Sep 12, 2017 |
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15928139 |
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14880776 |
Oct 12, 2015 |
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15701791 |
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14449328 |
Aug 1, 2014 |
9175018 |
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14880776 |
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13667164 |
Nov 2, 2012 |
8859504 |
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14449328 |
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12485344 |
Jun 16, 2009 |
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13667164 |
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61211499 |
Mar 31, 2009 |
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61132244 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P 31/00 20180101;
A61K 31/454 20130101; C07F 5/04 20130101; A61P 19/02 20180101; A61P
29/00 20180101; C07F 5/025 20130101; A61P 11/00 20180101; A61P
17/02 20180101; C07K 5/06139 20130101; A61P 1/04 20180101; A61P
9/10 20180101; A61P 1/00 20180101; A61P 7/06 20180101; A61P 1/16
20180101; A61P 35/00 20180101; A61P 17/06 20180101; A61P 27/02
20180101; A61P 25/00 20180101; A61P 25/28 20180101; A61P 35/04
20180101; A61P 37/06 20180101; A61P 9/00 20180101; A61P 11/06
20180101; A61K 31/198 20130101; A61P 17/00 20180101; C07C 233/83
20130101; A61P 25/14 20180101; A61P 35/02 20180101; A61P 25/16
20180101; C07K 5/06191 20130101; A61P 25/04 20180101; A61P 37/02
20180101; A61P 43/00 20180101; A61K 31/69 20130101; A61P 31/18
20180101 |
International
Class: |
C07F 5/04 20060101
C07F005/04; A61K 31/69 20060101 A61K031/69; A61K 31/454 20060101
A61K031/454; A61K 31/198 20060101 A61K031/198; C07C 233/83 20060101
C07C233/83; C07K 5/06 20060101 C07K005/06; C07F 5/02 20060101
C07F005/02; C07K 5/078 20060101 C07K005/078 |
Claims
1. A process for generating a compound of Formula (II) ##STR00018##
or a pharmaceutically acceptable salt thereof, wherein A is 0, 1,
or 2; P is R.sup.c--C(O)--; R.sup.c is R.sup.D; R.sup.D is
substituted or unsubstituted mono- or bicyclic ring system selected
from the group consisting of phenyl, pyridinyl, pyrimidinyl,
naphthyl, benzimidazolyl, quinolinyl, isoquinolinyl, quinoxalinyl,
tetrahydroquinolinyl, tetrahydroisoquinolinyl,
tetrahydroquinoxalinyl, and dihydrobenzoxazinyl; R.sup.a is
hydrogen, C.sub.1-6 aliphatic, C.sub.1-6 fluoroaliphatic,
--(CH.sub.2).sub.mCH.sub.2--R.sup.B,
--(CH.sub.2).sub.mCH.sub.2--NHC(.dbd.NR.sup.4)NH--Y,
--(CH.sub.2).sub.mCH.sub.2--CON(R.sup.4).sub.2,
--(CH.sub.2).sub.mCH.sub.2--N(R.sup.4)CON(R.sup.4).sub.2,
--(CH.sub.2).sub.mCH(R.sup.6)N(R.sup.4).sub.2,
--(CH.sub.2).sub.mCH(R.sup.5a)--OR.sup.5b, or
--(CH.sub.2).sub.mCH(R.sup.5)--SR.sup.S; R.sup.a1 is hydrogen,
C.sub.1-6 aliphatic, C.sub.1-6 fluoroaliphatic,
--(CH.sub.2).sub.mCH.sub.2--R.sup.B,
--(CH.sub.2).sub.mCH.sub.2--NHC(.dbd.NR.sup.4)NH--Y,
--(CH.sub.2).sub.mCH.sub.2--CON(R.sup.4).sub.2,
--(CH.sub.2).sub.mCH.sub.2--N(R.sup.4)CON(R.sup.4).sub.2,
--(CH.sub.2).sub.mCH(R.sup.6)N(R.sup.4).sub.2,
--(CH.sub.2).sub.mCH(R.sup.5a)--OR.sup.5b, or
--(CH.sub.2).sub.mCH(R.sup.5)--SR.sup.5; each R independently is
hydrogen, C.sub.1-6 aliphatic, C.sub.1-6 fluoroaliphatic,
--(CH.sub.2).sub.mCH.sub.2--R.sup.B,
--(CH.sub.2).sub.mCH.sub.2--NHC(.dbd.NR.sup.4)NH--Y,
--(CH.sub.2).sub.mCH.sub.2--CON(R.sup.4).sub.2,
--(CH.sub.2).sub.mCH.sub.2--N(R.sup.4)CON(R.sup.4).sub.2,
--(CH.sub.2).sub.mCH(R.sup.6)N(R.sup.4).sub.2,
--(CH.sub.2).sub.mCH(R.sup.5a)--OR.sup.5b, or
--(CH.sub.2).sub.mCH(R.sup.5)--SR.sup.5; each R.sup.B independently
is a substituted or unsubstituted mono- or bicyclic ring system;
each R.sup.4 independently is hydrogen or a substituted or
unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl group;
or two R.sup.4 on the same nitrogen atom, taken 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 N, O, and S; each
R.sup.5 independently is hydrogen or a substituted or unsubstituted
aliphatic, aryl, heteroaryl, or heterocyclyl group; each R.sup.5a
independently is hydrogen or a substituted or unsubstituted
aliphatic, aryl, heteroaryl, or heterocyclyl group; each R.sup.5b
independently is hydrogen or a substituted or unsubstituted
aliphatic, aryl, heteroaryl, or heterocyclyl group; each R.sup.6
independently is a substituted or unsubstituted aliphatic, aryl, or
heteroaryl group; Y is hydrogen, --CN, or --NO.sub.2; m is 0, 1, or
2; each of R.sup.b1 and R.sup.b2 independently is hydrogen,
--CO.sub.2H, --OH, or a substituted or unsubstituted aliphatic,
aryl, heteroaryl, or heterocyclyl group; each of R.sup.b3 and
R.sup.b4 independently is hydrogen, --CO.sub.2H, or a substituted
or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl
group; or R.sup.b2 and R.sup.b4 are each independently hydrogen,
and R.sup.b1 and R.sup.b3, taken together with the carbon atoms to
which they are attached, form an unsubstituted or substituted fused
4- to 8-membered non-aromatic ring having 0-3 ring heteroatoms
selected from the group consisting of O, N, and S, wherein said
ring may be optionally fused to an unsubstituted or substituted 4-
to 8-membered non-aromatic ring, or 5- to 6-membered aromatic ring
having 0-3 ring heteroatoms selected from the group consisting of
O, N, and S; or R.sup.b2 and R.sup.b4 are absent, and R.sup.b1 and
R.sup.b3, taken together with the carbon atoms to which they are
attached, form an unsubstituted or substituted fused 5- to
6-membered aromatic ring having 0-3 ring heteroatoms selected from
the group consisting of O, N, and S, wherein said ring may be
optionally fused to an unsubstituted or substituted 4- to
8-membered non-aromatic ring, or 5- to 6-membered aromatic ring
having 0-3 ring heteroatoms selected from the group consisting of
O, N, and S; and n is 0 or 1; the process comprising: (1) coupling
a compound of Formula (1) with a compound of Formula (ii) to form a
compound of Formula (iii) ##STR00019## wherein X'' is a counter
anion; and PG is hydrogen or a protecting group; (2) deprotecting
the compound of Formula (iii) to form a compound of Formula (iiia)
##STR00020## (3) coupling the compound of Formula (iiia) with a
compound, P-LG, to form a compound of Formula (iv) ##STR00021##
wherein LG is a leaving group; (4) deprotecting the compound of
Formula (iv) to form the compound of Formula (v) ##STR00022## and
(5) reacting the compound of Formula (v) with an alpha hydroxy
carboxylic acid or a beta hydroxy carboxylic acid to form a
compound of Formula (II).
2. The process of claim 1, wherein the reaction of step (1) or step
(3) or both is conducted in the presence of a peptide coupling
reagent.
3. The process of claim 2, wherein the peptide coupling reagent is
selected from the group consisting of a carbodiimide reagent,
phosphonium reagent, and uronium reagent.
4. The process of claim 3, wherein the peptide coupling reagent is
selected from one or more of the group consisting of
dicyclohexylcarbodiimide (DCC),
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC),
benzotriazol-1-yloxytris(dimethylamino)phosphonium
hexafluorophosphate (BOP), and
O-(1H-benzotriazol-1-yl)-N,N',N'-tetramethyluronium
tetrafluoroborate (TBTU).
5. The process of claim 1, further comprising converting the
carboxylic acid moiety of compound (ii) to an activated ester or
acid halide prior to the reaction of step (1).
6. The process of claim 5, wherein said activated ester or acid
halide is an O--(N-hydroxysuccinnimide) ester.
7. The process of claim 1, wherein the reaction of step (3) is
conducted in the presence of a solvent.
8. The process of claim 7, wherein the solvent is
tetrahydrofuran.
9. The process of claim 1, wherein the reaction of step (4) is
conducted in the presence of an aqueous mineral acid.
10. The process of claim 9, wherein the mineral acid is
hydrochloric acid.
11. The process of claim 1, wherein the reaction of step (4) is
conducted in the presence of an organic boronic acid acceptor.
12. The process of claim 11, wherein the organic boronic acid
acceptor is i-BuB(OH).sub.2.
13. The process of claim 1, wherein the reaction of step (5) is
conducted in the presence of a solvent selected from the group
consisting of ethyl acetate, methyl isobutyl ketone, acetone,
acetonitrile, 2-methyltetrahydrofuran, anisole, isopropyl acetate,
dimethoxyethane, tetrahydrofuran, dioxane, dichloromethane,
toluene, heptane, methyl-cyclohexane, tert-butylmethyl ether, and
any combination thereof.
14. The process of claim 1, wherein the reaction of step (5) is
conducted in the presence of a catalyst, wherein the catalyst is an
organic amine base.
15. The process of claim 14, wherein the catalyst is selected from
the group consisting of 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-diazabicycle[4.3.0]non-5-ene,
N,N'diisopropylethylamine, or any combination thereof.
16. The process of claim 1, wherein step (5) further comprises
cooling the solution.
17. The process of claim 16, wherein the cooling comprises cooling
the solution uncontrolled until the internal temperature is about
25.degree. C.
18. The process of claim 1, wherein the reaction of step (5) is
conducted in the presence of a co-solvent.
19. The process of claim 18, wherein the co-solvent is selected
from the group consisting of heptane, methylcyclohexane, toluene,
tert-butylmethyl ether, ethyl acetate, or any combination
thereof.
20. The process of claim 1, further comprising (6) isolating the
compound of Formula (II) as a crystalline solid.
21. The process of claim 1, wherein PG is a protecting group
selected from the group consisting of an acyl protecting group and
a urethane protecting group.
22. The process of claim 21, wherein PG is a protecting group
selected from the group consisting of formyl, acetyl, succinyl,
methoxysuccinyl, tert-butoxycarbonyl (Boc), benzyloxycarbonyl
(Cbz), and fluorenylmethoxycarbonyl (Fmoc).
23. The process of claim 1, wherein LG is --Cl.
24. The process of claim 1, wherein X.sup.- is
CF.sub.3CO.sub.2.sup.-.
25. The process of claim 1, wherein R.sup.D is
2,5-dichlorophenyl.
26. The process of claim 1, wherein A is 0; R.sup.a is C.sub.1-6
aliphatic; R.sup.a1 is hydrogen, C.sub.1-6 aliphatic,
--(CH.sub.2).sub.mCH.sub.2--R.sup.B, or
--(CH.sub.2).sub.mCH(R.sup.5a)--OR.sup.5b; each of R.sup.b1 and
R.sup.b2 independently is hydrogen, --CO.sub.2H, --OH, or a
substituted or unsubstituted aliphatic; each of R.sup.b3 and
Rb.sup.4 independently is hydrogen, C.sub.1-6 aliphatic, or
--(CH.sub.2).sub.pCO.sub.2H; and p is 0 or 1.
27. The process of claim 1, wherein A is 0; R.sup.D is
2,5-dichlorophenyl; R.sup.a is isobutyl; R.sup.a1 is hydrogen; each
R.sup.b1 and R.sup.b2 independently is hydrogen; each R.sup.b3 and
R.sup.b4 independently is --(CH.sub.2).sub.p--CO.sub.2H; p is 0 or
1; and n is 0 or 1.
28. The process of claim 1, wherein the alpha hydroxy carboxylic
acid or a beta hydroxy carboxylic acid is citric acid.
29. The process of claim 1, wherein the compound of Formula (II) is
4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-m-
ethylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid.
30. The process of claim 1, wherein the compound of Formula (II) is
2,2'-{2-[(1R)-1-({[(2,5-dichlorobenzoyl)amino]acetyl}amino)-3-methylbutyl-
]-5-oxo-1,3,2-dioxaborolane-4,4-diyl}diacetic acid.
31. A process for generating a compound of Formula (II)
##STR00023## or a pharmaceutically acceptable salt thereof, wherein
A is 0, 1, or 2; P is R.sup.c--C(O)--; R.sup.c is R.sup.D; R.sup.D
is substituted or unsubstituted mono- or bicyclic ring system
selected from the group consisting of phenyl, pyridinyl,
pyrimidinyl, naphthyl, benzimidazolyl, quinolinyl, isoquinolinyl,
quinoxalinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl,
tetrahydroquinoxalinyl, and dihydrobenzoxazinyl; R.sup.a is
hydrogen, C.sub.1-6 aliphatic, C.sub.1-6 fluoroaliphatic,
--(CH.sub.2).sub.mCH.sub.2R.sup.B,
--(CH.sub.2).sub.mCH.sub.2--NHC(.dbd.NR.sup.4)NH--Y,
--(CH.sub.2).sub.mCH.sub.2--CON(R.sup.4).sub.2,
--(CH.sub.2).sub.mCH.sub.2--N(R.sup.4)CON(R.sup.4).sub.2,
--(CH.sub.2).sub.mCH(R.sup.6)N(R.sup.4).sub.2,
--(CH.sub.2).sub.mCH(R.sup.5a)--OR.sup.5b, or
--(CH.sub.2).sub.mCH(R.sup.5)--SR.sup.5; R.sup.a1 is hydrogen,
C.sub.1-6 aliphatic, C.sub.1-6 fluoroaliphatic,
--(CH.sub.2).sub.mCH.sub.2R.sup.B,
--(CH.sub.2).sub.mCH.sub.2--NHC(.dbd.NR.sup.4)NH--Y,
--(CH.sub.2).sub.mCH.sub.2--CON(R.sup.4).sub.2,
--(CH.sub.2).sub.mCH.sub.2--N(R.sup.4)CON(R.sup.4).sub.2,
--(CH.sub.2).sub.mCH(R.sup.6)N(R.sup.4).sub.2,
--(CH.sub.2).sub.mCH(R.sup.5a)--OR.sup.5b, or
--(CH.sub.2).sub.mCH(R.sup.5)--SR.sup.5; each R independently is
hydrogen, C.sub.1-6 aliphatic, C.sub.1-6 fluoroaliphatic,
--(CH.sub.2).sub.mCH.sub.2R.sup.B,
--(CH.sub.2).sub.mCH.sub.2--NHC(.dbd.NR.sup.4)NH--Y,
--(CH.sub.2).sub.mCH.sub.2.gtoreq.CON(R.sup.4).sub.2,
--(CH.sub.2).sub.mCH.sub.2--N(R.sup.4)CON(R.sup.4).sub.2,
--(CH.sub.2).sub.mCH(R.sup.6)N(R.sup.4).sub.2,
--(CH.sub.2).sub.mCH(R.sup.5a)--OR.sup.5b, or
--(CH.sub.2).sub.mCH(R.sup.5)--SR.sup.5; each R.sup.B independently
is a substituted or unsubstituted mono- or bicyclic ring system;
each R.sup.4 independently is hydrogen or a substituted or
unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl group;
or two R.sup.4 on the same nitrogen atom, taken 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 N, O, and S; each
R.sup.5 independently is hydrogen or a substituted or unsubstituted
aliphatic, aryl, heteroaryl, or heterocyclyl group; each R.sup.5a
independently is hydrogen or a substituted or unsubstituted
aliphatic, aryl, heteroaryl, or heterocyclyl group; each R.sup.5b
independently is hydrogen or a substituted or unsubstituted
aliphatic, aryl, heteroaryl, or heterocyclyl group; each R.sup.6
independently is a substituted or unsubstituted aliphatic, aryl, or
heteroaryl group; Y is hydrogen, --CN, or --NO.sub.2; m is 0, 1, or
2; each of R.sup.b1 and R.sup.b2 independently is hydrogen,
--CO.sub.2H, --OH, or a substituted or unsubstituted aliphatic,
aryl, heteroaryl, or heterocyclyl group; each of R.sup.b3 and
R.sup.b4 independently is hydrogen, --CO.sub.2H, or a substituted
or unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl
group; or R.sup.b2 and R.sup.b4 are each independently hydrogen,
and R.sup.b1 and R.sup.b3, taken together with the carbon atoms to
which they are attached, form an unsubstituted or substituted fused
4- to 8-membered non-aromatic ring having 0-3 ring heteroatoms
selected from the group consisting of O, N, and S, wherein said
ring may be optionally fused to an unsubstituted or substituted 4-
to 8-membered non-aromatic ring, or 5- to 6-membered aromatic ring
having 0-3 ring heteroatoms selected from the group consisting of
O, N, and S; or R.sup.b2 and R.sup.b4 are absent, and R.sup.b1 and
R.sup.b3, taken together with the carbon atoms to which they are
attached, form an unsubstituted or substituted fused 5- to
6-membered aromatic ring having 0-3 ring heteroatoms selected from
the group consisting of O, N, and S, wherein said ring may be
optionally fused to an unsubstituted or substituted 4- to
8-membered non-aromatic ring, or 5- to 6-membered aromatic ring
having 0-3 ring heteroatoms selected from the group consisting of
O, N, and S; and n is 0 or 1; the process comprising: (1a) coupling
a compound of Formula (vi) with a compound, P-LG, to form a
compound of Formula (vii) ##STR00024## wherein PG is hydrogen or a
protecting group; and LG is a leaving group; (2a) deprotecting the
compound of Formula (vii) to form a compound of Formula (viia)
##STR00025## (3a) coupling the compound of Formula (viia) with a
compound of Formula (i) to form a compound of Formula (iv)
##STR00026## wherein X.sup.- is a counter anion; (4) deprotecting
the compound of Formula (iv) to form a compound of Formula (v)
##STR00027## and (5) reacting the compound of Formula (v) with an
alpha hydroxy carboxylic acid or a beta hydroxy carboxylic acid to
form a compound of Formula (II).
Description
PRIORITY
[0001] This application is a divisional of U.S. Utility patent
application Ser. No. 14/449,328 filed Aug. 1, 2014, which is a
divisional of U.S. Utility patent application Ser. No. 13/667,164,
filed Nov. 2, 2012, which is a continuation of U.S. utility patent
application Ser. No. 12/485,344, filed Jun. 16, 2009, which claims
priority from U.S. Provisional Patent Application Ser. No.
61/132,244, filed Jun. 17, 2008, and U.S. Provisional Patent
Application Ser. No. 61/211,499, filed Mar. 31, 2009; both which
are hereby incorporated by reference in their entirety.
FIELD OF THE INVENTION
[0002] The present invention relates to boronate ester compounds
useful as proteasome inhibitors. The invention also provides
pharmaceutical compositions comprising the compounds of the
invention, and methods of using the compositions in the treatment
of various diseases.
BACKGROUND OF THE INVENTION
[0003] Boronic acid and boronate ester compounds display a variety
of pharmaceutically useful biological activities. Shenvi et al.,
U.S. Pat. No. 4,499,082 (1985), discloses that peptide boronic
acids are inhibitors of certain proteolytic enzymes. Kenner and
Shenvi, U.S. Pat. No. 5,187,157 (1993), U.S. Pat. No. 5,242,904
(1993), and U.S. Pat. No. 5,250,720 (1993), describe a class of
peptide boronic acids that inhibit trypsin-like proteases. Kleeman
et al., U.S. Pat. No. 5,169,841 (1992), discloses N-terminally
modified peptide boronic acids that inhibit the action of renin.
Kinder et al., U.S. Pat. No. 5,106,948 (1992), discloses that
certain boronic acid compounds inhibit the growth of cancer cells.
Magde et al., WO 04/022070 discloses peptide boronic acid compounds
that inhibit thrombin. Boucher, U.S. Patent Application Pub. No.
2006/0084592 discloses various basic addition salts of peptide
boronic acid compounds. Bachovchin et al., WO 07/005991, discloses
peptide boronic acid compounds that inhibit fibroblast activating
protein.
[0004] Boronic acid and ester compounds hold particular promise as
inhibitors of the proteasome, a multicatalytic protease responsible
for the majority of intracellular protein turnover. Adams et al.,
U.S. Pat. No. 5,780,454 (1998), describes peptide boronic ester and
acid compounds useful as proteasome inhibitors. The reference also
describes the use of boronic ester and acid compounds to reduce the
rate of muscle protein degradation, to reduce the activity of
NF-.kappa.B in a cell, to reduce the rate of degradation of p53
protein in a cell, to inhibit cyclin degradation in a cell, to
inhibit the growth of a cancer cell, and to inhibit NF-.kappa.B
dependent cell adhesion. Furet et al., WO 02/096933, Chatterjee et
al., WO 05/016859, and Bernadini et al, WO 05/021558 and WO
06/08660, disclose additional boronic ester and acid compounds that
are reported to have proteasome inhibitory activity.
[0005] Ciechanover, Cell, 79:13-21 (1994), discloses that the
proteasome is the proteolytic component of the ubiquitin-proteasome
pathway, in which proteins are targeted for degradation by
conjugation to multiple molecules of ubiquitin. Ciechanover also
discloses that the ubiquitin-proteasome pathway plays a key role in
a variety of important physiological processes. Rivett et al.,
Biochem. 1 291:1 (1993) discloses that the proteasome displays
tryptic-, chymotryptic-, and peptidylglutamyl-peptidase activities.
Constituting the catalytic core of the 26S proteasome is the 20S
proteasome. McCormack et al., Biochemistry 37:7792 (1998), teaches
that a variety of peptide substrates, including
Suc-Leu-Leu-Val-Tyr-AMC, Z-Leu-Leu-Arg-AMC, and Z-Leu-Leu-Glu-2NA,
wherein Suc is N-succinyl, AMC is 7-amino-4-methylcoumarin, and 2NA
is 2-naphthylamine, are cleaved by the 20S proteasome.
[0006] Proteasome inhibition represents an important new strategy
in cancer treatment. King et al., Science 274:1652-1659 (1996),
describes an essential role for the ubiquitin-proteasome pathway in
regulating cell cycle, neoplastic growth and metastasis. The
authors teach that a number of key regulatory proteins, including,
cyclins, and the cyclin-dependent kinases p21 and p27.sup.KIP1, are
temporally degraded during the cell cycle by the
ubiquitin-proteasome pathway. The ordered degradation of these
proteins is required for the cell to progress through the cell
cycle and to undergo mitosis.
[0007] Furthermore, the ubiquitin-proteasome pathway is required
for transcriptional regulation. Palombella et al., Cell, 78:773
(1994), teaches that the activation of the transcription factor
NF-.kappa.B is regulated by proteasome-mediated degradation of the
inhibitor protein IKB. In turn, NF-.kappa.B plays a central role in
the regulation of genes involved in the immune and inflammatory
responses. Read et al., Immunity 2:493-506 (1995), teaches that the
ubiquitin-proteasome pathway is required for 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 extravastation
of tumor cells to and from the vasculature to distant tissue sites
within the body. Moreover, Beg and Baltimore, Science 274:782
(1996), teaches that NF-.kappa.B is an anti-apoptotic controlling
factor, and inhibition of NF-.kappa.B activation makes cells more
sensitive to environmental stress and cytotoxic agents.
[0008] The proteasome inhibitor VELCADE.RTM. (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), reviews the
clinical studies leading to the approval of bortezomib for the
treatment of multiple myeloma patients who have received at least
one prior therapy. Fisher et al., J. Clin. Oncol., 30:4867 (2006),
describes an international multi-center Phase II study 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 activities of bortezomib.
[0009] Structural analysis reported by Voges et al., Annu. Rev.
Biochem., 68:1015 (1999) reveals that the 20S proteasome comprises
28 subunits, with the catalytic subunits .beta.1, .beta.2, and
.beta.5 being responsible for peptidylglutamyl, tryptic, and
chymotryptic peptidase activity, respectively. Rivett et al., Curr.
Protein Pept. Sci., 5:153 (2004) discloses that when the proteasome
is exposed to certain cytokines, including IFN-.gamma. and
TNF-.alpha., the .beta.1, .beta.2, and .beta.5 subunits are
replaced with alternate catalytic subunits, .beta.1i, .beta.2i, and
.beta.5i, to form a variant form of the proteasome known as the
immunoproteasome.
[0010] Orlowski, Hematology (Am. Soc. Hematol. Educ. Program) 220
(2005), discloses that the immunoproteasome also is expressed
constitutively in some cells derived from hematopoietic precursors.
The author suggests that inhibitors specific for the
immunoproteasome may allow for targeted therapy against cancers
arising from hematologic origins, thereby potentially sparing
normal tissues, such as gastrointestinal and neurological 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 generate 1-butanol and boric acid. These
difficulties limit the pharmaceutical utility of boronic acid
compounds, complicating the characterization of pharmaceutical
agents comprising boronic acid compounds and limiting their
shelf-life.
[0012] Plamondon et al., WO 02/059131 discloses stable,
pharmaceutically acceptable compositions prepared from boronic acid
compounds and sugars. There remains a need for additional stable
formulations of boronic acid compounds.
BRIEF DESCRIPTION OF THE DRAWINGS
[0013] FIG. 1 is a powder X-ray diffractogram of
4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-m-
ethylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid (I-1) Form
1.
[0014] FIG. 2 is a differential scanning calorimetry (DSC)/thermal
gravimetric analysis (TGA) profile for 4-(R,
S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-methyl-
butyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid (I-1) Form
1.
[0015] FIG. 3 is a powder X-ray diffractogram of
4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-m-
ethylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid (I-1) Form
2.
[0016] FIG. 4 is a differential scanning calorimetry (DSC)/thermal
gravimetric analysis (TGA) profile for
4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-m-
ethylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid (I-1) Form
2.
[0017] FIG. 5 is a powder X-ray diffractogram of
2,5-dichloro-N-[2-({(1R)-3-methyl-1-[(4S)-4-methyl-5-oxo-1,3,2-dioxaborol-
an-2-yl]butyl.BECAUSE.amino)-2-oxoethyl]benzamide (I-7).
[0018] FIG. 6 is a powder X-ray diffractogram of
2,5-dichloro-N-(2-{[(1R)-3-methyl-1-(4-oxo-4H-1,3,2-benzodioxaborinin-2-y-
l)butyl}amino)-2-oxoethyl)benzamide (I-13).
[0019] FIG. 7 is a powder X-ray diffractogram of
4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-m-
ethylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid (I-1) Form
2.
[0020] FIG. 8 is a differential scanning calorimetry (DSC) profile
of
4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-m-
ethylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid (I-1) Form
2.
DESCRIPTION OF THE INVENTION
[0021] The present invention provides novel boronate ester
compounds and stable pharmaceutically acceptable compositions
comprising them. These compounds and compositions are useful for
inhibiting proteasome activity in vitro and in vivo, and are
especially useful for the treatment of various cell proliferative
diseases.
[0022] In one aspect, the invention provides compounds of of the
general formula (I):
##STR00001##
[0023] or a pharmaceutically acceptable salt or thereof, wherein:
[0024] A is 0, 1, or 2; [0025] P is hydrogen or an
amino-group-blocking moiety; [0026] R.sup.a is hydrogen, C.sub.1-6
aliphatic, C.sub.1-6 fluoroaliphatic,
--(CH.sub.2).sub.m--CH.sub.2--R.sup.B,
--(CH.sub.2).sub.m--CH.sub.2--NHC(.dbd.NR.sup.4)NH--Y,
--(CH.sub.2).sub.m--CH.sub.2--CON(R.sup.4).sub.2,
--(CH.sub.2).sub.m--CH.sub.2--N(R.sup.4)CON(R.sup.4).sub.2,
--(CH.sub.2).sub.m--CH(R.sup.6)N(R.sup.4).sub.2,
--(CH.sub.2).sub.m--CH(R.sup.5a)--OR.sup.5b, or
--(CH.sub.2).sub.m--CH(R.sup.5)--SR.sup.5; [0027] R.sup.a1 is
hydrogen, C.sub.1-6 aliphatic, C.sub.1-6 fluoroaliphatic,
--(CH.sub.2).sub.m--CH.sub.2--R.sup.B,
--(CH.sub.2).sub.m--CH.sub.2--NHC(.dbd.NR.sup.4)NH--Y,
--(CH.sub.2).sub.m--CH.sub.2--CON(R.sup.4).sub.2,
--(CH.sub.2).sub.m--CH.sub.2--N(R.sup.4)CON(R.sup.4).sub.2,
--(CH.sub.2).sub.m--CH(R.sup.6)N(R.sup.4).sub.2,
--(CH.sub.2).sub.m--CH(R.sup.5a)--OR.sup.5b, or
--(CH.sub.2).sub.m--CH(R.sup.5)--SR.sup.5; [0028] each R.sup.a2
independently is hydrogen, C.sub.1-6 aliphatic, C.sub.1-6
fluoroaliphatic, --(CH.sub.2).sub.m--CH.sub.2--R.sup.B,
--(CH.sub.2).sub.m--CH.sub.2--NHC(.dbd.NR.sup.4)NH--Y,
--(CH.sub.2).sub.m--CH.sub.2--CON(R.sup.4).sub.2,
--(CH.sub.2).sub.m--CH.sub.2--N(R.sup.4)CON(R.sup.4).sub.2,
--(CH.sub.2).sub.m--CH(R.sup.6)N(R.sup.4).sub.2,
--(CH.sub.2).sub.m--CH(R.sup.5a)--OR.sup.5B, or
--(CH.sub.2).sub.m--CH(R.sup.5)--SR.sup.5; [0029] each R.sup.B
independently is a substituted or unsubstituted mono- or bicyclic
ring system; [0030] each R.sup.4 independently is hydrogen or a
substituted or unsubstituted aliphatic, aryl, heteroaryl, or
heterocyclyl group; or two R.sup.4 on the same nitrogen atom, taken
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; [0031] each
R.sup.5 independently is hydrogen or a substituted or unsubstituted
aliphatic, aryl, heteroaryl, or heterocyclyl group; [0032] each
R.sup.5a independently is hydrogen or a substituted or
unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl group;
[0033] each R.sup.5b independently is hydrogen or a substituted or
unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl group;
[0034] each R.sup.6 independently is a substituted or unsubstituted
aliphatic, aryl, or heteroaryl group; [0035] Y is hydrogen, --CN,
or --NO.sub.2; [0036] m is 0, 1, or 2; and [0037] Z.sup.1 and
Z.sup.2 together form a moiety derived from an alpha-hydroxy
carboxylic acid, wherein the atom attached to boron in each case is
an oxygen atom; or Z.sup.1 and Z.sup.2 together form a moiety
derived from a beta-hydroxy carboxylic acid, wherein the atom
attached to boron in each case is an oxygen atom.
[0038] In another aspect, the present invention provides
pharmaceutical compositions comprising the compound of formula (I),
or a crystalline form thereof, and additional excipients described
herein, suitable for the production of an oral pharmaceutical
dosage form.
[0039] In another aspect, the invention provides a pharmaceutical
composition comprising the compound of formula (I), or a
crystalline form thereof, and additional excipients described
herein, suitable for the production of a lyophilized powder
pharmaceutical dosage form.
[0040] In another aspect, the invention provides a pharmaceutical
composition comprising the compound of formula (I), or a
crystalline form thereof, and additional excipients described
herein, suitable for the production of a liquid pharmaceutical
dosage form.
[0041] In another aspect, the invention provides a pharmaceutical
composition, comprising the compound of formula (I), or a
crystalline form thereof, a filler, and optionally a lubricant.
[0042] In another aspect, the invention provides a pharmaceutical
composition, comprising the compound of formula (I), or a
crystalline form thereof, a filler, optionally a lubricant,
optionally a flow-aid, and optionally a buffer.
[0043] In another aspect, the invention provides a pharmaceutical
composition, comprising the compound of formula (I), or a
crystalline form thereof, a bulking agent, and a buffer.
[0044] In another aspect, the invention provides processes for the
production of the pharmaceutical compositions of the invention.
[0045] In another aspect, the invention provides methods for the
use of the pharmaceutical compositions of the invention, for
treating a patient having, or at risk of developing or experiencing
a recurrence of, a proteasome-mediated disorder.
[0046] In another aspect, the invention provides methods for the
use of the pharmaceutical compositions of the invention for the
treatment of cancer.
Definitions
[0047] Unless otherwise explicitly stated, the term "proteasome" is
intended to refer to both constitutive proteasome and
immunoproteasome.
[0048] The term "aliphatic" or "aliphatic group", as used herein,
means a substituted or unsubstituted straight-chain, branched, or
cyclic C.sub.1-12 hydrocarbon, which is completely saturated or
which contains one or more units of unsaturation, but which is not
aromatic. For example, suitable aliphatic groups include
substituted or unsubstituted linear, branched or cyclic alkyl,
alkenyl, or alkynyl groups and hybrids thereof, such as
(cycloalkyl)alkyl, (cycloalkenyl)alkyl or (cycloalkyl)alkenyl. In
various embodiments, the aliphatic group has 1 to 12, 1 to 8, 1 to
6, 1 to 4, or 1 to 3 carbons.
[0049] The terms "alkyl", "alkenyl", and "alkynyl", used alone or
as part of a larger moiety, refer to a straight or branched chain
aliphatic group having from 1 to 12 carbon atoms. For purposes of
the present invention, the term "alkyl" will be used when the
carbon atom attaching the aliphatic group to the rest of the
molecule is a saturated carbon atom. However, an alkyl group may
include unsaturation at other carbon atoms. Thus, alkyl groups
include, without limitation, methyl, ethyl, propyl, allyl,
propargyl, butyl, pentyl, and hexyl.
[0050] For purposes of the present invention, the term "alkenyl"
will 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, without limitation, vinyl, 1-propenyl,
1-butenyl, 1-pentenyl, and 1-hexenyl.
[0051] For purposes of the present invention, the term "alkynyl"
will 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, without limitation, ethynyl, 1-propynyl,
1-butynyl, 1-pentynyl, and 1-hexynyl.
[0052] The term "cycloaliphatic", used alone or as part of a larger
moiety, refers to a saturated or partially unsaturated cyclic
aliphatic ring system having from 3 to about 14 members, wherein
the aliphatic ring system is optionally substituted. In some
embodiments, the cycloaliphatic is a monocyclic hydrocarbon having
3-8 or 3-6 ring carbon atoms. Nonlimiting examples include
cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl,
cyclohexenyl, cycloheptyl, cycloheptenyl, cyclooctyl, cyclooctenyl,
and cyclooctadienyl. In some embodiments, the cycloaliphatic is a
bridged or fused bicyclic hydrocarbon having 6-12, 6-10, or 6-8
ring carbon atoms, wherein any individual ring in the bicyclic ring
system has 3-8 members.
[0053] In some embodiments, two adjacent substituents on the
cycloaliphatic ring, taken together with the intervening ring
atoms, form an optionally substituted fused 5- to 6-membered
aromatic or 3- to 8-membered non-aromatic ring having 0-3 ring
heteroatoms selected from the group consisting of O, N, and S.
Thus, the term "cycloaliphatic" includes aliphatic rings that are
fused to one or more aryl, heteroaryl, or heterocyclyl rings.
Nonlimiting examples include indanyl,
5,6,7,8-tetrahydroquinoxalinyl, decahydronaphthyl, or
tetrahydronaphthyl, where the radical or point of attachment is on
the aliphatic ring.
[0054] The terms "aryl" and "ar-", used alone or as part of a
larger moiety, e.g., "aralkyl", "aralkoxy", or "aryloxyalkyl",
refer to a C.sub.6 to C.sub.1-4 aromatic hydrocarbon, comprising
one to three rings, each of which is optionally substituted.
Preferably, the aryl group is a C.sub.6-10 aryl group. Aryl groups
include, without limitation, phenyl, naphthyl, and anthracenyl. In
some embodiments, two adjacent substituents on the aryl ring, taken
together with the intervening ring atoms, form an optionally
substituted fused 5- to 6-membered aromatic or 4- to 8-membered
non-aromatic ring having 0-3 ring heteroatoms selected from the
group consisting of O, N, and S. Thus, the term "aryl", as used
herein, includes groups in which an aryl ring is fused to one or
more heteroaryl, cycloaliphatic, or heterocyclyl rings, where the
radical or point of attachment is on the aromatic ring. Nonlimiting
examples of such fused ring systems include indolyl, isoindolyl,
benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl,
benzimidazolyl, benzthiazolyl, quinolyl, isoquinolyl, cinnolinyl,
phthalazinyl, quinazolinyl, quinoxalinyl, carbazolyl, acridinyl,
phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl,
tetrahydroisoquinolinyl, fluorenyl, indanyl, phenanthridinyl,
tetrahydronaphthyl, indolinyl, phenoxazinyl, benzodioxanyl, and
benzodioxolyl. An aryl group may be mono-, bi-, tri-, or
polycyclic, preferably mono-, bi-, or tricyclic, more preferably
mono- or bicyclic. The term "aryl" may be used interchangeably with
the terms "aryl group", "aryl moiety", and "aryl ring".
[0055] An "aralkyl" or "arylalkyl" group comprises an aryl group
covalently attached to an alkyl group, either of which
independently is optionally substituted. Preferably, the aralkyl
group is C.sub.6-10 aryl(C.sub.1-6)-alkyl, C.sub.6-10
aryl(C.sub.1-4)alkyl, or C.sub.6-10 aryl(C.sub.1-3)alkyl,
including, without limitation, benzyl, phenethyl, and
naphthylmethyl.
[0056] 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; having 6, 10, or 14 .pi. electrons shared in a
cyclic array; and having, in addition to carbon atoms, from one to
four heteroatoms. The term "heteroatom" refers to nitrogen, oxygen,
or sulfur, and includes any oxidized form of nitrogen or sulfur,
and any quaternized form of a basic nitrogen. Thus, when used in
reference to a ring atom of a heteroaryl, the term "nitrogen"
includes an oxidized nitrogen (as in pyridine N-oxide). Certain
nitrogen atoms of 5-membered heteroaryl groups also are
substitutable, as further defined below. Heteroaryl groups include,
without limitation, 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" means a monovalent radical
produced by removal of a hydrogen radical from the parent
heteroaromatic ring system. The radical (i.e., the point of
attachment of the heteroaryl to the rest of the molecule) may be
created at any substitutable position on any ring of the parent
heteroaryl ring system.
[0057] In some embodiments, two adjacent substituents on the
heteroaryl, taken together with the intervening ring atoms, form an
optionally substituted fused 5- to 6-membered aromatic or 4- to
8-membered non-aromatic ring having 0-3 ring heteroatoms selected
from the group consisting of O, N, and S. Thus, the terms
"heteroaryl" and "heteroar-", 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. Nonlimiting examples
include indolyl, isoindolyl, benzothienyl, benzofuranyl,
dibenzofuranyl, indazolyl, benzimidazolyl, benzthiazolyl,
benzoxazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl,
quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl,
phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl,
tetrahydroisoquinolinyl, and pyrido[2,3-b]-1,4-oxazin-3(4H)-one. A
heteroaryl group may be mono-, bi-, tri-, or polycyclic, preferably
mono-, bi-, or tricyclic, more preferably mono- or bicyclic. The
term "heteroaryl" may be used interchangeably with the terms
"heteroaryl ring", or "heteroaryl group", any of which terms
include rings that are optionally substituted. The term
"heteroaralkyl" refers to an alkyl group substituted by a
heteroaryl, wherein the alkyl and heteroaryl portions independently
are optionally substituted.
[0058] As used herein, the terms "aromatic ring" and "aromatic ring
system" refer to an optionally substituted mono-, bi-, or tricyclic
group having 0-6, preferably 0-4 ring heteroatoms, and having 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.
[0059] As used herein, the terms "heterocycle", "heterocyclyl",
"heterocyclic radical", and "heterocyclic ring" are used
interchangeably and refer to a stable 3- to 7-membered monocyclic,
or to a fused 7- to 10-membered or bridged 6- to 10-membered
bicyclic heterocyclic moiety that is either saturated or partially
unsaturated, and having, in addition to carbon atoms, one or more,
preferably one to four, heteroatoms, as defined above. When used in
reference to a ring atom of a heterocycle, the term "nitrogen"
includes a substituted nitrogen. As an example, in a heterocyclyl
ring having 1-3 heteroatoms selected from the group consisting of
oxygen, sulfur or nitrogen, the nitrogen may be N (as in
3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl), or .sup.-NR (as
in N-substituted pyrrolidinyl). A heterocyclic ring can be attached
to its pendant group at any heteroatom or carbon atom that results
in a stable structure, and any of the ring atoms can be optionally
substituted. Examples of such saturated or partially unsaturated
heterocyclic radicals include, without limitation,
tetrahydrofuranyl, tetrahydrothienyl, pyrrolidinyl, pyrrolidonyl,
piperidinyl, pyrrolinyl, tetrahydroquinolinyl,
tetrahydroisoquinolinyl, decahydroquinolinyl, oxazolidinyl,
piperazinyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl,
thiazepinyl, morpholinyl, and quinuclidinyl.
[0060] In some embodiments, two adjacent substituents on a
heterocyclic ring, taken together with the intervening ring atoms,
form an optionally substituted fused 5- to 6-membered aromatic or
3- to 8-membered non-aromatic ring having 0-3 ring heteroatoms
selected from the group consisting of O, N, and S. Thus, the terms
"heterocycle", "heterocyclyl", "heterocyclyl ring", "heterocyclic
group", "heterocyclic moiety", and "heterocyclic radical", are used
interchangeably herein, and include groups in which a heterocyclyl
ring is fused to one or more aryl, heteroaryl, or cycloaliphatic
rings, such as indolinyl, 3H-indolyl, chromanyl, phenanthridinyl,
or tetrahydroquinolinyl, where the radical or point of attachment
is on the heterocyclyl ring. A heterocyclyl group may be mono-,
bi-, tri-, or polycyclic, preferably mono-, bi-, or tricyclic, more
preferably mono- or bicyclic. The term "heterocyclylalkyl" refers
to an alkyl group substituted by a heterocyclyl, wherein the alkyl
and heterocyclyl portions independently are optionally
substituted.
[0061] As used herein, the term "partially unsaturated" refers to a
ring moiety that includes at least one double or triple bond
between ring atoms. The term "partially unsaturated" is intended to
encompass rings having multiple sites of unsaturation, but is not
intended to include aryl or heteroaryl moieties, as herein
defined.
[0062] The terms "haloaliphatic", "haloalkyl", "haloalkenyl" and
"haloalkoxy" refer to an aliphatic, alkyl, alkenyl or alkoxy group,
as the case may be, which is substituted with one or more halogen
atoms. As used herein, the term "halogen" or "halo" means F, Cl,
Br, or I. The term "fluoroaliphatic" refers to a haloaliphatic
wherein the halogen is fluoro, including perfluorinated aliphatic
groups. Examples of fluoroaliphatic groups include, without
limitation, fluoromethyl, difluoromethyl, trifluoromethyl,
2-fluoroethyl, 2,2,2-trifluoroethyl, 1,1,2-trifluoroethyl,
1,2,2-trifluoroethyl, and pentafluoroethyl.
[0063] The term "linker group" or "linker" means an organic moiety
that connects two parts of a compound. Linkers typically comprise
an atom such as oxygen or sulfur, a unit such as --NH--,
--CH.sub.2--, --C(O)--, --C(O)NH--, or a chain of atoms, such as an
alkylene chain. The molecular mass of a linker is typically in the
range of about 14 to 200, preferably in the range of 14 to 96 with
a length of up to about six atoms. In some embodiments, the linker
is a C.sub.1-6 alkylene chain.
[0064] The term "alkylene" refers to a bivalent alkyl group. An
"alkylene chain" is a polymethylene group, i.e.,
--(CH.sub.2).sub.y-, wherein y is a positive integer, preferably
from 1 to 6, from 1 to 4, from 1 to 3, from 1 to 2, or from 2 to 3.
A substituted alkylene chain is a polymethylene group in which one
or more methylene hydrogen atoms is replaced with a substituent.
Suitable substituents include those described below for a
substituted aliphatic group. An alkylene chain also may be
substituted at one or more positions with an aliphatic group or a
substituted aliphatic group.
[0065] An alkylene chain also can be optionally interrupted by a
functional group. An alkylene chain is "interrupted" by a
functional group when an internal methylene unit is replaced with
the functional group. Examples of suitable "interrupting functional
groups" include --C(R*).dbd.C(R*)--, --C.ident.C--, --O--, --S--,
--S(O)--, --S(O).sub.2--, --S(O).sub.2N(R.sup.+)--, --N(R*)--,
--N(R.sup.+)CO--, --N(R.sup.+)C(O)N(R.sup.+)--,
--N(R.sup.+)C(.dbd.NR.sup.+)--N(R.sup.+)--,
--N(R.sup.+)--C(.dbd.NR.sup.+)--, --N(R.sup.+)CO.sub.2--,
--N(R.sup.+)SO.sub.2--, --N(R.sup.+)SO.sub.2N(R.sup.+)--,
--OC(O)--, --OC(O)O--, --OC(O)N(R.sup.+)--, --C(O)--, --CO.sub.2--,
--C(O)N(R.sup.+)--, --C(O)--C(O)--,
--C(.dbd.NR.sup.+)--N(R.sup.+)--, --C(NR.sup.+).dbd.N--,
--C(.dbd.NR.sup.+)--O--, --C(OR*).dbd.N--,
--C(R.sup.o)--.dbd.N--O--, or --N(R.sup.+)--N(R.sup.+)--. Each
R.sup.+ independently is hydrogen or an optionally substituted
aliphatic, aryl, heteroaryl, or heterocyclyl group, or two R.sup.+
on the same nitrogen atom, taken 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* independently is
hydrogen or an optionally substituted aliphatic, aryl, heteroaryl,
or heterocyclyl group. Each R.sup.o independently is an optionally
substituted aliphatic, aryl, or heteroaryl group.
[0066] Examples of C.sub.3-6 alkylene chains that have been
"interrupted" with --O-- include --CH.sub.2OCH.sub.2--,
--CH.sub.2O(CH.sub.2).sub.2--, --CH.sub.2O(CH.sub.2).sub.3--,
--CH.sub.2O(CH.sub.2).sub.4--, --(CH.sub.2).sub.2OCH.sub.2--,
--(CH.sub.2).sub.2O(CH.sub.2).sub.2--, --(CH.sub.2).sub.2O
(CH.sub.2).sub.3--, --(CH.sub.2).sub.3O(CH.sub.2)--,
--(CH.sub.2).sub.3O(CH.sub.2).sub.2--, and
--(CH.sub.2).sub.4O(CH.sub.2)--. Other examples of alkylene chains
that are "interrupted" with functional groups include
--CH.sub.2Z*CH.sub.2--, --CH.sub.2Z*(CH.sub.2).sub.2--,
--CH.sub.2Z*(CH.sub.2).sub.3--, --CH.sub.2Z*(CH.sub.2).sub.4--,
--(CH.sub.2).sub.2Z*CH.sub.2--,
--(CH.sub.2).sub.2Z*(CH.sub.2).sub.2--,
--(CH.sub.2).sub.2Z*(CH.sub.2).sub.3--,
--(CH.sub.2).sub.3Z*(CH.sub.2)--,
--(CH.sub.2).sub.3Z*(CH.sub.2)--.sub.2--, and
--(CH.sub.2).sub.4Z*(CH.sub.2)--, wherein Z* is one of the
"interrupting" functional groups listed above.
[0067] One of ordinary skill in the art will recognize that when an
alkylene chain having an interruption is attached to a functional
group, certain combinations would 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 which maintains its integrity
long enough to be useful for therapeutic or prophylactic
administration to a patient. Preferably, the chemical structure is
not substantially altered when kept at a temperature below
-70.degree. C., below -50.degree. C., below -20.degree. C., below
0.degree. C., or below 20.degree. C., in the absence of moisture or
other chemically reactive conditions for at least a week.
[0068] The term "substituted", as used herein, means that a
hydrogen radical of the designated 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 designated atom, means
that attached to the atom is a hydrogen radical, which can be
replaced with the radical of a suitable substituent.
[0069] The phrase "one or more substituents", as used herein,
refers to a number of substituents that equals from one to the
maximum number of substituents possible based on the number of
available bonding sites, provided that the above conditions of
stability and chemical feasibility are met. Unless otherwise
indicated, an optionally substituted group may have a substituent
at each substitutable position of the group, and the substituents
may be either the same or different.
[0070] As used herein, the terms "independently" or "independently
selected" means that the same or different values may be selected
for multiple instances of a given variable in a single
compound.
[0071] An aryl (including the aryl moiety in aralkyl, aralkoxy,
aryloxyalkyl and the like) or heteroaryl (including the heteroaryl
moiety in heteroaralkyl and heteroaralkoxy and the like) group may
contain one or more substituents. Examples of suitable substituents
on the unsaturated carbon atom of an aryl or heteroaryl group
include -halo, --NO.sub.2,--CN, --R*, --C(R*).dbd.C(R*).sub.2,
--C--R*, --OR*, --SR.sup.o, --S(O)R.sup.o, --SO.sub.2R.sup.o,
--SO.sub.3R*, --SO.sub.2N(R.sup.+).sub.2, --N(R.sup.+).sub.2,
--NR.sup.+C(O)R*, --NR.sup.+C(O)N(R.sup.+).sub.2,
--N(R.sup.+)C(.dbd.NR.sup.+)--N(R.sup.+).sub.2,
--N(R.sup.+)C(.dbd.NR.sup.+)--R.sup.o, --NR.sup.+CO.sub.2R.sup.o,
--NR.sup.+SO.sub.2R.sup.o, --NR.sup.+SO.sub.2N(R.sup.+).sub.2,
--O--C(O)R*, --O--CO.sub.2R*, --OC(O)N(R.sup.+).sub.2, --C(O)R*,
--CO.sub.2R*, --C(O)--C(O)R*, --C(O)N(R.sup.+).sub.2,
--C(O)N(R.sup.+)--OR*,
--C(O)N(R.sup.+)C(.dbd.NR.sup.+)--N(R.sup.+).sub.2,
--N(R.sup.+)C(.dbd.NR.sup.+)--N(R.sup.+)--C(O)R*,
--C(.dbd.NR.sup.+)--N(R.sup.+).sub.2, --C(.dbd.NR.sup.+)--OR*,
--N(R.sup.+)--N(R.sup.+).sub.2,
--C(.dbd.NR.sup.+)--N(R.sup.+)--OR*, --C(R.sup.o).dbd.N--OR*,
--P(O)(R*).sub.2, --P(O)(OR*).sub.2, --O--P(O)--OR*, and
--P(O)(NR.sup.+)--N(R.sup.+).sub.2, wherein R.sup.o, R.sup.+, and
R* are as defined above, or two adjacent substituents, taken
together with their 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.
[0072] An aliphatic group or a non-aromatic heterocyclic ring may
be substituted with one or more substituents. Examples of suitable
substituents on the saturated carbon of an aliphatic group or of a
non-aromatic heterocyclic ring include, without limitation, those
listed above for the unsaturated carbon of an aryl or heteroaryl
group and the following: .dbd.O, .dbd.S, .dbd.C(R*).sub.2,
.dbd.N--N(R*).sub.2, .dbd.N--OR*, .dbd.N--NHC(O)R*,
.dbd.N--NHCO.sub.2R.sup.o, .dbd.N--NHSO.sub.2R.sup.o, or
.dbd.N--R*, where each R* and R.sup.o is as defined above.
[0073] Suitable substituents on a substitutable nitrogen atom of a
heteroaryl or non-aromatic heterocyclic ring include --R*,
--N(R*).sub.2, --C(O)R*, --CO.sub.2R*,
--C(O)--C(O)R*--C(O)CH.sub.2C(O)R*, --SO.sub.2R*,
--SO.sub.2N(R*).sub.2, --C(.dbd.S)N(R*).sub.2,
--C(.dbd.NH)--N(R*).sub.2, and --NR*SO.sub.2R*; wherein each R* is
as defined above. A ring nitrogen atom of a heteroaryl or
non-aromatic heterocyclic ring also may be oxidized to form the
corresponding N-hydroxy or N-oxide compound. A nonlimiting example
of such a heteroaryl having an oxidized ring nitrogen atom is
N-oxidopyridyl.
[0074] The term "about" is used herein to mean approximately, in
the region of, roughly, or around. When the term "about" is used in
conjunction with a numerical range, it modifies that range by
extending the boundaries above and below the numerical values set
forth. In general, the term "about" is used herein to modify a
numerical value above and below the stated value by a variance of
10%.
[0075] As used herein, the term "comprises" means "includes, but is
not limited to."
[0076] It will be apparent to one skilled in the art that certain
compounds of this invention may exist in tautomeric forms, all such
tautomeric forms of the compounds being within the scope of the
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, as well as all stereochemical forms of the
structure; i.e., the R and S configurations for each asymmetric
center. Therefore, single stereochemical isomers as well as
enantiomeric and diastereomeric mixtures of the present compounds
are within the scope of the invention. When a mixture is enriched
in one stereoisomer relative to another stereoisomer, the mixture
may contain, for example, an enantiomeric excess of at least 50%,
75%, 90%, 99%, or 99.5%.
[0077] Unless otherwise stated, structures depicted herein are also
meant to include compounds which differ only in the presence of one
or more isotopically enriched atoms. For example, compounds having
the present structure except for the replacement of a hydrogen atom
by a deuterium or tritium, or the replacement of a carbon atom by a
.sup.13C- or .sup.14C-enriched carbon are within the scope of the
invention.
[0078] As used herein, the term "seeding" is used to refer to the
addition of a crystalline material to initiate crystallization or
recrystallization.
[0079] When a compound crystallizes from a solution or slurry, it
may crystallize with different spatial lattice arrangements, a
property referred to as "polymorphism." Each of the crystal forms
is a "polymorph." While polymorphs of a given substance have the
same chemical composition, 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, flow properties, and/or solid state
stability.
[0080] As used herein, the term "solvate or solvated" means a
physical association of a compound with one or more solvent
molecules. This physical association includes hydrogen bonding. In
certain instances the solvate will be capable of isolation, for
example when one or more solvent molecules are incorporated in the
crystal lattice of the crystalline solid. "Solvate or solvated"
encompasses both solution-phase and isolable solvates.
Representative solvates include, for example, hydrates,
ethanolates, or methanolates. The physical properties of a solvate
typically 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 "pseudo-polymorphs".
[0081] As used herein, the term "hydrate" is a solvate wherein the
solvent molecule is H.sub.2O that is present in a defined
stoichiometric amount, and may, for example, include hemihydrate,
monohydrate, dihydrate, or trihydrate. As used herein, the term
"anhydrate" is a compound of the invention that contains no
H.sub.2O incorporated in its crystal lattice.
[0082] As used herein, "crystalline" refers to a solid having a
highly regular chemical structure. In particular, a crystalline
compound may be produced as one or more single crystalline forms of
the compound. For the purposes of this application, the terms
"single crystalline form" or "crystalline form" are used
interchangeably and distinguish between crystals that have
different properties (e.g., different XRPD patterns, different DSC
scan results). Thus, each distinct polymorph and pseudopolymorph of
a compound is considered to be a distinct single crystalline form
herein.
[0083] "Substantially crystalline" refers to a compound that may be
at least a particular weight percent crystalline. Particular weight
percentages are 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 compounds that are
at least 70% crystalline. In other embodiments, substantially
crystalline refers to compounds that are at least 90%
crystalline.
[0084] "Substantially pure" refers to a compound that may be at
least a particular weight percent of the compound. Particular
weight percentages are 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%.
[0085] Unless otherwise explicitly stated, structures depicted
herein are meant to include all hydrates, anhydrates, solvates and
polymorphs thereof
[0086] 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" are used
interchangeably, and include all crystalline forms. Both terms
refer to the compounds produced in Example 1 and Example 1A in the
Examples below including both Form 1 and Form 2.
[0087] 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"
are used interchangeably. Both terms refer to the crystalline form
2 produced in Example 1 Form 2 and Example 1A in the Examples
below.
[0088] As used herein, the terms "compound of formula (VIII-1)",
and "(R)-1-((2,5-dichlorobenzamido)acetamido)-3-methylbutylboronic
acid" are used interchangeably. The compound of formula (VIII-1) is
disclosed in U.S. Pat. No. 7,442,830 and WO 09/020448.
[0089] As used herein, the terms "compound of formula (I-15)",
"compound (I-15)" and "(I-15)" are used interchangeably and are
used to refer to the citrate ester of the compound (VIII-15), and
the compound produced in Example 15 of the Examples below.
[0090] As used herein, the term "anhydride" used in reference to a
boronic acid such as the compound of formula (VIII), refers to a
chemical compound formed by combination of two or more molecules of
a boronic acid compound, with loss of one or more water molecules.
When mixed with water, the boronic acid anhydride compound is
hydrated to release the free boronic acid compound. In various
embodiments, the boronic acid anhydride can comprise two, three,
four, or more boronic acid units, and can have a cyclic or linear
configuration. Non-limiting examples of oligomeric boronic acid
anhydrides of peptide boronic acids compound of the invention are
illustrated below:
##STR00002##
[0091] In formula (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 comprises a cyclic trimer
("boroxine") of formula (2), wherein nn is 1. The variable W has
the formula (3):
##STR00003##
wherein P, R.sup.a2, A, R.sup.a1 and R.sup.a are as defined
herein.
[0092] As used herein, the total weight of a single oral
pharmaceutical dosage form is determined by adding all the weights
of the components in the oral pharmaceutical dosage form, and does
not include the weight of any coatings which may be optionally
applied to the oral pharmaceutical dosage form after it is formed.
The total weight of a single oral pharmaceutical dosage form is
used as the basis for calculating the weight percentage of each of
the components that comprise the oral pharmaceutical dosage
form.
[0093] As used herein, "low-moisture" used in reference to an
excipient such as a filler, refers to an excipient that has a water
content of about 0.5% to about 4%. The term "low-moisture" may be
used interchangeably with the term "low-water".
[0094] As used herein, the term "lyophilized powder", "cake", or
"lyophilized cake" refers to any solid material obtained by
lyophilization of an aqueous mixture.
[0095] As used herein, the term "tonicity modifier" refers to
agents which contribute to the osmolality of a liquid or
solution.
[0096] As used herein, the terms "boronate ester" and "boronic
ester" are used interchangeably and refer to a chemical compound
containing a --B(Z.sup.1)(Z.sup.2) moiety, wherein Z.sup.1 and
Z.sup.2 together form a moiety where the atom attached to boron in
each case is an oxygen atom.
[0097] In some embodiments, the boronate ester moiety is a
5-membered ring. In some other embodiments, the boronate ester
moiety is a 6-membered ring. In some other embodiments, the
boronate ester moiety is a mixture of a 5-membered ring and a
6-membered ring.
[0098] As used herein, the term "alpha-hydroxy carboxylic acid"
refers to a compound that contains a hydroxyl group directly
attached to a carbon atom in an alpha position relative to a
carboxylic acid group. As used herein, the term "alpha-hydroxy
carboxylic acid" is not intended to be limited to compounds having
only one hydroxyl group and one carboxylic acid group.
[0099] As used herein, the term "beta-hydroxy carboxylic acid"
refers to a compound that contains a hydroxyl group directly
attached to a carbon atom in a beta position relative to a
carboxylic acid group. As used herein, the term "beta-hydroxy
carboxylic acid" is not intended to be limited to compounds having
only one hydroxyl group and one carboxylic acid group.
[0100] As used herein, the term "moiety derived from an
alpha-hydroxy carboxylic acid" refers to a moiety formed by
removing a hydrogen atom from a carboxylic acid within an
alpha-hydroxy carboxylic acid and by removing a hydrogen atom from
a hydroxyl group directly attached to a carbon atom in an alpha
position relative to the carboxylic acid group. As used herein, the
term "moiety derived from a beta-hydroxy carboxylic acid" refers to
a moiety formed by removing a hydrogen atom from a carboxylic acid
within a beta-hydroxy carboxylic acid and by removing a hydrogen
atom from a hydroxyl group directly attached to a carbon atom in a
beta position relative to the carboxylic acid group.
DETAILED DESCRIPTION OF THE INVENTION
[0101] In some embodiments the alpha-hydroxy acid is characterized
by formula (V):
##STR00004## [0102] wherein each of R.sup.b3 and R.sup.b4
independently is hydrogen, --CO.sub.2H, or a substituted or
unsubstituted aliphatic, aryl, heteroaryl or heterocyclyl
group.
[0103] In some embodiments, each of R.sup.b3 and R.sup.b4
independently is hydrogen, C.sub.1-6 aliphatic, or
--(CH.sub.2).sub.p--CO.sub.2H, and p is 0, 1 or 2. In some
embodiments, each of R.sup.b3 and R.sup.b4 independently is
hydrogen or C.sub.1-6 aliphatic. In certain such embodiments, each
of R.sup.b3 and R.sup.b4 independently is selected from the group
consisting of hydrogen, methyl, ethyl, isopropyl, isobutyl,
tert-butyl, and cyclohexyl. In some other embodiments, each of
R.sup.b3 and R.sup.b4 independently is hydrogen or
--(CH.sub.2).sub.p--CO.sub.2H. In some such embodiments, p is 1. In
certain other embodiments, each of R.sup.b3 and R.sup.b4
independently is --(CH.sub.2).sub.p--CO.sub.2H. In certain such
embodiments, p is 1.
[0104] In some embodiments, the alpha-hydroxy carboxylic 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 alpha-hydroxy
carboxylic 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
certain embodiments, the alpha-hydroxy carboxylic acid is citric
acid. Some other non-limiting examples of alpha-hydroxy carboxylic
acids include glucoheptonic acid, maltonic acid, lactobionic acid,
and galactaric acid.
[0105] In some embodiments the beta-hydroxy acid is characterized
by formula (VI):
##STR00005## [0106] wherein each of R.sup.b1 and R.sup.b2
independently is hydrogen, --CO.sub.2H, --OH, or a substituted or
unsubstituted aliphatic, aryl, heteroaryl or heterocyclyl group;
each of R.sup.b3 and R.sup.b4 independently is hydrogen,
--CO.sub.2H, or a substituted or unsubstituted aliphatic, aryl,
heteroaryl or heterocyclyl group; [0107] or R.sup.b2 and R.sup.b4
are each independently hydrogen, and R.sup.b1 and R.sup.b3, taken
together with the carbon atoms to which they are attached, form an
unsubstituted or substituted 4- to 8-membered non-aromatic ring
having 0-3 ring heteroatoms selected from the group consisting of
O, N, and S, wherein said ring may optionally be fused to an
unsubstituted or substituted 4- to 8-membered non-aromatic ring or
5- to 6-membered aromatic ring having 0-3 ring heteroatoms selected
from the group consisting of O, N, and S; [0108] or R.sup.b2 and
R.sup.b4 are absent, and R.sup.b1 and R.sup.b3, taken together with
the carbon atoms to which they are attached, form an unsubstituted
or substituted 5- to 6-membered aromatic ring having 0-3 ring
heteroatoms selected from the group consisting of O, N, and S,
wherein said ring may optionally be fused to an unsubstituted or
substituted 4- to 8-membered non-aromatic ring, or 5- to 6-membered
aromatic ring having 0-3 ring heteroatoms selected from the group
consisting of O, N, and S.
[0109] In some embodiments each of R.sup.b1 and R.sup.b2
independently is hydrogen, C.sub.1-6 aliphatic,
--(CH.sub.2).sub.p--OH, or --(CH.sub.2).sub.p--CO.sub.2H, and p is
0, 1 or 2. In some such embodiments, each of R.sup.b1 and R.sup.b2
is hydrogen. In some other such embodiments, R.sup.b1 is --OH and
R.sup.b2 is hydrogen.
[0110] In some embodiments each of R.sup.b3 and R.sup.b4
independently is hydrogen, C.sub.1-6 aliphatic, or
--(CH.sub.2).sub.p--CO.sub.2H, and p is 0, 1 or 2. In some
embodiments, each of R.sup.b3 and R.sup.b4 independently is
hydrogen or C.sub.1-6 aliphatic. In certain such embodiments, each
of R.sup.b3 and R.sup.b4 independently is selected from the group
consisting of hydrogen, methyl, ethyl, isopropyl, isobutyl,
tert-butyl, and cyclohexyl. In certain other embodiments, each of
R.sup.b3 and R.sup.b4 independently is
--(CH.sub.2).sub.p--CO.sub.2H, and p is 0 or 1.
[0111] The variable p is 0, 1, or 2. In some embodiments, p is 0 or
1. In certain embodiments, p is 0. In other certain embodiments, p
is 1.
[0112] In some embodiments, R.sup.b2 and R.sup.b4 are absent and
R.sup.b1 and R.sup.b3 taken together with the carbon atoms to which
they are attached, form a substituted or unsubstituted phenyl
ring.
[0113] In some embodiments, the beta-hydroxy carboxylic acid is
selected from the group consisting of malic acid, citric acid,
3-hydroxybutyric acid, beta-hydroxyisovaleric acid, and salicylic
acid. In some other embodiments, the beta-hydroxy carboxylic acid
is selected from the group consisting of malic acid, citric acid,
3-hydroxybutyric acid, beta-hydroxyisovaleric acid, tartaric acid,
and salicylic acid. In certain embodiments, the beta-hydroxy
carboxylic acid is citric acid. Some other non-limiting examples of
beta-hydroxy carboxylic acids include glucoheptonic acid, maltonic
acid, lactobionic acid, and galactaric acid. Some other
non-limiting examples of beta-hydroxy carboxylic acids include
embonic acid, 1-hydroxy-2-naphthoic acid and 3-hydroxy-2-naphthoic
acid.
[0114] In some embodiments, the alpha-hydroxy acid or beta-hydroxy
acid is selected from the group consisting of glycolic acid, malic
acid, hexahydromandelic acid, 2-hydroxyisobutyric acid, citric
acid, mandelic acid, lactic acid, 3-hydroxybutyric acid,
beta-hydroxyisovaleric acid, 2-hydroxy-3,3-dimethylbutyric acid,
2-hydroxy-3-methylbutyric acid, 2-hydroxyisocaproic acid, tartaric
acid, salicylic acid, and benzilic acid.
[0115] In some embodiments, compounds of general formula (I) are
characterized by formula (II):
##STR00006## [0116] wherein: [0117] the variables P, A, R.sup.a,
R.sup.a1, R.sup.a2, and n have the values described below and the
variables R.sup.b1, R.sup.b2, R.sup.b3 and R.sup.b4 have the values
described above.
[0118] In some embodiments, any one of R.sup.b1, R.sup.b2, R.sup.b3
and R.sup.b4 may contain a functional group that can form a further
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.
[0119] In some embodiments, wherein the alpha-hydroxy carboxylic
acid or beta-hydroxy carboxylic acid is citric acid, the compound
of general formula (I) is characterized by formula (III) or
(IV):
##STR00007##
[0120] or a mixture thereof, wherein the variables P, A, R.sup.a,
R.sup.a1, and R.sup.a2 have the values described below.
[0121] In some other embodiments, wherein the alpha-hydroxy
carboxylic acid or beta-hydroxy carboxylic acid is citric acid, a
further bond can be formed between the carboxylic acid in formula
(III) or (IV) and the boron atom. Without being limited by any
chemical bonding theory, in such embodiments, the compound of
general formula (I) may be represented by formula (IIIa) or
(IVa):
##STR00008##
[0122] or a mixture thereof, wherein the variables P, A, R.sup.a,
R.sup.a1, and R.sup.a2 have the values described below.
[0123] It is recognized that, without being limited by any chemical
bonding theory, there are other representations that could be used
to depict this further bonding of the carboxylic acid with the
boron atom in formulas (IIIa) and (IVa).
[0124] The following values are described for the variables in any
of formulas (I), (II), (III), (IIIa), (IV), or (IVa).
[0125] The variable P is hydrogen or an amino-group-blocking
moiety. Non-limiting examples of amino-group-blocking moieties can
be found in P. G. M. Wuts and T. W. Greene, Greene's Protective
Groups in Organic Synthesis (4.sup.th ed.), John Wiley & Sons,
NJ (2007), and include, e.g., acyl, sulfonyl, oxyacyl, and
aminoacyl groups.
[0126] In some embodiments, P is R.sup.c--C(O)--,
R.sup.c--O--C(O)--, R.sup.c13 N(R.sup.4c)--C(O)--,
R.sup.c--S(O).sub.2--, or R.sup.c--N(R.sup.4c)--S(O).sub.2--, where
R.sup.c is selected from the group consisting of C.sub.1-6
aliphatic, C.sub.1-6 fluoroaliphatic, --R.sup.D, -T.sup.1-R.sup.D,
and, -T.sup.1-R.sup.2c, and the variables T.sup.1, R.sup.D,
R.sup.2c and R.sup.4c, have the values described below.
[0127] The variable R.sup.4c is hydrogen, C.sub.1-4 alkyl,
C.sub.1-4 fluoroalkyl, or C.sub.6-10 ar(C.sub.1-4 alkyl), the aryl
portion of which is substituted or unsubstituted. In some
embodiments, R.sup.4c is hydrogen or C.sub.1-4 alkyl. In certain
embodiments, R.sup.4c is hydrogen.
[0128] The variable T.sup.1 is a C.sub.1-6 alkylene chain
substituted with 0-2 independently selected R.sup.3a or R.sup.3b,
wherein the alkylene chain optionally is interrupted by
--C(R.sup.5).dbd.C(R.sup.5)--, --C.quadrature.C--, or --O--. Each
R.sup.3a independently is selected from the group consisting of
--F, --OH, --O(C.sub.1-4 alkyl), --CN, --N(R.sup.4).sub.2,
--C(O)--(C.sub.1-4 alkyl), --CO.sub.2H, --CO.sub.2(C.sub.1-4
alkyl), --C(O)NH.sub.2, and --C(O)--NH(C.sub.1-4 alkyl). Each
R.sup.3b independently is a C.sub.1-3 aliphatic optionally
substituted with R.sup.3a or R.sup.7; or two substituents R.sup.3b
on the same carbon atom, taken together with the carbon atom to
which they are attached, form a 3- to 6-membered cycloaliphatic
ring. Each R.sup.7 is a substituted or unsubstituted aromatic
group. In some embodiments, T.sup.1 is a C.sub.1-4 alkylene
chain.
[0129] The variable R.sup.2c is halo, --OR.sup.5, --SR.sup.6,
--S(O)R.sup.6, --SO.sub.2R.sup.6, --SO.sub.2N(R.sup.4).sub.2,
--N(R.sup.4).sub.2, --NR.sup.4C(O)R.sup.5,
--NR.sup.4C(O)N(R.sup.4).sub.2, --NR.sup.4CO.sub.2R.sup.6,
--N(R.sup.4)SO.sub.2R.sup.6, --N(R.sup.4)SO.sub.2N(R.sup.4).sub.2,
--O--C(O)R.sup.5, --OC(O)N(R.sup.4).sub.2, --C(O)R.sup.5,
--CO.sub.2R.sup.5, or --C(O)N(R.sup.4).sub.2, where:
[0130] each R.sup.4 independently is hydrogen or an optionally
substituted aliphatic, aryl, heteroaryl, or heterocyclyl group; or
two R.sup.4 on the same nitrogen atom, taken together with the
nitrogen atom, form an optionally substituted 4- to 8-membered
heterocyclyl ring having, in addition to the nitrogen atom, 0-2
ring heteroatoms independently selected from the group consisting
of N, O, and S;
[0131] each R.sup.5 independently is hydrogen or an optionally
substituted aliphatic, aryl, heteroaryl, or heterocyclyl group;
and
[0132] each R.sup.6 independently is an optionally substituted
aliphatic, aryl, or heteroaryl group.
[0133] The variable R.sup.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.sup.D
is substituted on substitutable ring carbon atoms with 0-2 R.sup.d
and 0-2 R.sup.8d, and each substitutable ring nitrogen atom in
R.sup.D is unsubstituted or is substituted with --C(O)R.sup.5,
--C(O)N(R.sup.4).sub.2, --CO.sub.2R.sup.6, --SO.sub.2R.sup.6,
--SO.sub.2N(R.sup.4).sub.2, C.sub.1-4 aliphatic, a substituted or
unsubstituted C.sub.6-10 aryl, or a C.sub.6-10 ar(C.sub.1-4alkyl,
the aryl portion of which is substituted or unsubstituted. The
variables R.sup.4, R.sup.5, and R.sup.6 have the values described
above. Each R.sup.d independently is selected from the group
consisting of C.sub.1-6 aliphatic, C.sub.1-6 fluoroaliphatic, halo,
--R.sup.1d, --R.sup.2d, -T.sup.2-R.sup.1d, and -T.sup.2-R.sup.2d,
where the variables T.sup.2, R.sup.1d, R.sup.2d, and R.sup.8d have
the values described below. In some embodiments, each R.sup.d
independently is selected from the group consisting of C.sub.1-6
aliphatic, C.sub.1-6 fluoroaliphatic and halo.
[0134] T.sup.2 is a C.sub.1-6 alkylene chain substituted with 0-2
independently selected R.sup.3a or R.sup.3b, wherein the alkylene
chain optionally is interrupted by --C(R.sup.5).dbd.C(R.sup.5)--,
--C.ident.C--, or --O---. The variables R.sup.3a, R.sup.3b, and
R.sup.5 have the values described above.
[0135] Each R.sup.1d independently is a substituted or
unsubstituted aryl, heteroaryl, heterocyclyl, or cycloaliphatic
ring.
[0136] Each R.sup.2d independently is --NO.sub.2, --CN,
--C(R.sup.5).dbd.C(R.sup.5).sub.2, --OR.sup.5, --SR.sup.6,
--S(O)R.sup.6, --SO.sub.2R.sup.6, --SO.sub.2N(R.sup.4).sub.2,
--N(R.sup.4).sub.2, --NR.sup.4C(O)R.sup.5,
--NR.sup.4C(O)N(R.sup.4).sub.2,
--N(R.sup.4)C(.dbd.NR.sup.4)--N(R.sup.4).sub.2,
--N(R.sup.4)C(.dbd.NR.sup.4)--R.sup.6, --NR.sup.4CO.sub.2R.sup.6,
--N(R.sup.4)SO.sub.2R.sup.6, --N(R.sup.4)SO.sub.2N(R.sup.4).sub.2,
--O--C(O)R.sup.5, --OC(O)N(R.sup.4).sub.2, --C(O)R.sup.5,
--CO.sub.2R.sup.5, --C(O)N(R.sup.4).sub.2,
--C(O)N(R.sup.4)--OR.sup.5,
--C(O)N(R.sup.4)C(.dbd.NR.sup.4)--N(R.sup.4).sub.2,
--N(R.sup.4)C(.dbd.NR.sup.4)--N(R.sup.4)--C(O)R.sup.5, or
--C(.dbd.NR.sup.4)--N(R.sup.4).sub.2. The variables R.sup.4,
R.sup.5, and R.sup.6 have the values described above.
[0137] Each R.sup.8d independently is selected from the group
consisting of C.sub.1-4 aliphatic, C.sub.1-4 fluoroaliphatic, halo,
--OH, --O(C.sub.1-4 aliphatic), --NH.sub.2, --NH(C.sub.1-4
aliphatic), and --N(C.sub.1-4 aliphatic).sub.2. In some
embodiments, each R.sup.8d independently is C.sub.1-4 aliphatic,
C.sub.1-4 fluoroaliphatic or halo.
[0138] In some embodiments, R.sup.D is a substituted or
unsubstituted mono- or bicyclic ring system. In some embodiments
R.sup.D is a substituted or unsubstituted mono- 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. In some embodiments, R.sup.D is a substituted
or unsubstituted mono- or bicyclic ring system selected from the
group consisting of phenyl, pyridinyl, pyrimidinyl, pyrazinyl,
naphthyl, benzimidazolyl, quinolinyl, isoquinolinyl, quinoxalinyl,
tetrahydroquinolinyl, tetrahydroisoquinolinyl,
tetrahydroquinoxalinyl, and dihydrobenzoxazinyl.
[0139] In some embodiments, the substitutable ring carbon atoms in
R.sup.D are substituted on substitutable carbon atoms with 0-1
R.sup.d and 0-2 R.sup.8d; wherein:
[0140] each R.sup.d independently is C.sub.1-6 aliphatic, C.sub.1-6
fluoroaliphatic or halo; and
[0141] each R.sup.8d independently is C.sub.1-4 aliphatic,
C.sub.1-4 fluoroaliphatic or halo.
[0142] In some embodiments, the substitutable ring carbon atoms in
R.sup.D are substituted with 0-1 R.sup.d and 0-2 R.sup.8d,
wherein:
[0143] T.sup.1 is a C.sub.1-3 alkylene chain that is unsubstituted
or is substituted with R.sup.3a or R.sup.3b;
[0144] each R.sup.1d independently is a substituted or
unsubstituted aryl, heteroaryl, heterocyclyl, or cycloaliphatic
ring; and
[0145] each R.sup.2d independently is --OR.sup.5, --SR.sup.6,
--S(O)R.sup.6, --SO.sub.2R.sup.6, --SO.sub.2N(R.sup.4).sub.2,
--N(R.sup.4).sub.2, --NR.sup.4C(O)R.sup.5,
--NR.sup.4C(O)N(R.sup.4).sub.2, --O--C(O)R.sup.5,
--OC(O)N(R.sup.4).sub.2, --C(O)R.sup.5, --CO.sub.2R.sup.5, or
--C(O)N(R.sup.4).sub.2. The variables R.sup.4, R.sup.5, and R.sup.6
have the values described above.
[0146] In some embodiments, the variable R.sup.d has the formula
-Q--R.sup.E, where Q is --O--, --NH--, or --CH.sub.2--, and R.sup.E
is a substituted or unsubstituted aryl, heteroaryl, heterocyclyl,
or cycloaliphatic ring. In some embodiments, R.sup.E is a
substituted or unsubstituted phenyl, pyridinyl, pyrimidinyl,
pyrazinyl, piperidinyl, piperazinyl, or morpholinyl ring.
[0147] In some embodiments, P has the formula R.sup.c--C(O)--,
where R.sup.c is C.sub.1-4 alkyl, C.sub.1-4 fluoroalkyl, or
C.sub.6-10 ar(C.sub.1-4) alkyl, the aryl portion of which is
substituted or unsubstituted. In certain such embodiments, P is
selected from the group consisting of acetyl, trifluoroacetyl, and
phenylacetyl.
[0148] In some other embodiments, P has the formula
R.sup.D--C(O)--, where R.sup.D is a substituted or unsubstituted
phenyl, pyridinyl, pyrazinyl, pyrimidinyl, quinolinyl, or
quinoxalinyl. In yet some other embodiments, P has the formula
R.sup.D--C(O), where R.sup.D is a phenyl, pyridinyl, pyrazinyl,
pyrimidinyl, naphthyl, quinolinyl, quinoxalinyl, benzimidazolyl, or
dihydrobenzoxazinyl substituted with 0-1 R.sup.d and 0-2
R.sup.8d.
[0149] In certain embodiments, P has the formula R.sup.D--C(O)--,
where R.sup.D is 2-pyrazinyl. In other certain embodiments, P has
the formula R.sup.D--C(O)--, where R.sup.D is 2,5-dichlorophenyl.
In yet other certain embodiments, P has the formula
R.sup.D--C(O)--, where R.sup.D is 6-phenyl-2-pyridinyl.
[0150] In some other embodiments, P has the formula
R.sup.c--SO.sub.2--, where R.sup.c is --R.sup.D or
-T.sup.1-R.sup.D, where T.sup.1 is C.sub.1-4 alkylene and R.sup.D
is a phenyl, pyridinyl, pyrazinyl, pyrimidinyl, naphthyl,
quinolinyl, quinoxalinyl, benzimidazolyl, or dihydrobenzoxazinyl
substituted with 0-1 R.sup.d and 0-2 R.sup.8d.
[0151] The variable R.sup.a is hydrogen, C.sub.1-6 aliphatic,
C.sub.1-6 fluoroaliphatic, --(CH.sub.2).sub.m--CH.sub.2--R.sup.B,
--(CH.sub.2).sub.m--CH.sub.2--NHC(.dbd.NR.sup.4)NH--Y,
--(CH.sub.2).sub.m--CH.sub.2--CON(R.sup.4).sub.2,
--(CH.sub.2).sub.m--CH.sub.2--N(R.sup.4)CON(R.sup.4).sub.2,
--(CH.sub.2).sub.m--CH(R.sup.6)N(R.sup.4).sub.2,
--(CH.sub.2).sub.m--CH(R.sup.5a)--OR.sup.5b, or
--(CH.sub.2).sub.m--CH(R.sup.5)--SR.sup.5, where the variables
R.sup.4, R.sup.5, and R.sup.6 have the values described above, and
the variables R.sup.5a, R.sup.5b, R.sup.B, Y, and m have the values
described below.
[0152] In some embodiments, R.sup.a is hydrogen, C.sub.1-6
aliphatic, C.sub.1-6 fluoroaliphatic, or
--(CH.sub.2).sub.m--CH.sub.2--R.sup.B. In some other embodiments,
R.sup.a is C.sub.1-6 aliphatic, or
--(CH.sub.2).sub.m--CH.sub.2--R.sup.B. In some further embodiments,
R.sup.a is C.sub.1-6 aliphatic. In yet other further embodiments,
R.sup.a is isobutyl, 1-naphthylmethyl, 2-napththylmethyl, benzyl,
4-fluorobenzyl, 4-hydroxybenzyl, 4-(benzyloxy)benzyl,
benzylnapththylmethyl or phenethyl. In certain embodiments, R.sup.a
is isobutyl.
[0153] The variable R.sup.a1 is hydrogen, C.sub.1-6 aliphatic,
C.sub.1-6 fluoroaliphatic, --(CH.sub.2).sub.m--CH.sub.2--R.sup.B,
--(CH.sub.2).sub.m--CH.sub.2--NHC(.dbd.NR.sup.4)NH--Y,
--(CH.sub.2).sub.m--CH.sub.2--CON(R.sup.4).sub.2,
--(CH.sub.2).sub.m--CH.sub.2--N(R.sup.4)CON(R.sup.4).sub.2,
--(CH.sub.2).sub.m--CH(R.sup.6)N(R.sup.4).sub.2,
--(CH.sub.2).sub.m--CH(R.sup.5a)--OR.sup.5b, or
--(CH.sub.2).sub.m--CH(R.sup.5)--SR.sup.5, where the variables
R.sup.4, R.sup.5, and R.sup.6 have the values described above, and
the variables R.sup.5a, R.sup.5b, R.sup.B, Y, and m have the values
described below.
[0154] In some embodiments, R.sup.a1 is hydrogen, C.sub.1-6
aliphatic, C.sub.1-6 fluoroaliphatic,
--(CH.sub.2).sub.m--CH.sub.2--R.sup.B, or
--(CH.sub.2).sub.m--CH(R.sup.5a)--OR.sup.5b. In some other
embodiments, R.sup.a1 is hydrogen,
--(CH.sub.2).sub.m--CH.sub.2--R.sup.B, or
--(CH.sub.2).sub.m--CH(R.sup.5a)--OR.sup.5b. In yet some other
embodiments, R.sup.a1 is isobutyl, 1-naphthylmethyl,
2-naphthylmethyl, benzyl, 4-fluorobenzyl, 4-hydroxybenzyl,
4-(benzyloxy)benzyl, benzylnaphthylmethyl or phenethyl.
[0155] In certain embodiments, R.sup.a1 is --CH.sub.2--R.sup.B. In
other certain embodiments, R.sup.a1 is --CH(R.sup.5a)--OR.sup.5b.
In yet other certain embodiments, R.sup.a1 is hydrogen.
[0156] The variable R.sup.a2 is hydrogen, C.sub.1-6 aliphatic,
C.sub.1-6 fluoroaliphatic, --(CH.sub.2).sub.m--CH.sub.2--R.sup.B,
--(CH.sub.2).sub.m--CH.sub.2--NHC(.dbd.NR.sup.4)NH--Y,
--(CH.sub.2).sub.m--CH.sub.2--CON(R.sup.4).sub.2,
--(CH.sub.2).sub.m--CH.sub.2--N(R.sup.4)CON(R.sup.4).sub.2,
--(CH.sub.2).sub.m--CH(R.sup.6)N(R.sup.4).sub.2,
--(CH.sub.2).sub.m--CH(R.sup.5a)--OR.sup.5b, or
--(CH.sub.2).sub.m--CH(R.sup.5)--SR.sup.S, where the variables
R.sup.4, R.sup.5, and R.sup.6 have the values described above, and
the variables R.sup.B, R.sup.5a, R.sup.5b, Y, and m have the values
described below.
[0157] In some embodiments, R.sup.a2 is hydrogen, C.sub.1-6
aliphatic, C.sub.1-6 fluoroaliphatic,
--(CH.sub.2).sub.m--CH.sub.2--R.sup.B, or
--(CH.sub.2).sub.m--CH(R.sup.5a)--OR.sup.5b. In some other
embodiments, R.sup.a2 is isobutyl, 1-naphthylmethyl,
2-naphthylmethyl, benzyl, 4-fluorobenzyl, 4-hydroxybenzyl,
4-(benzyloxy)benzyl, benzylnaphthylmethyl or phenethyl.
[0158] Each R.sup.B, independently, is a substituted or
unsubstituted mono- or bicyclic ring system. In some embodiments,
each R.sup.B independently is a substituted or unsubstituted
phenyl, pyridyl, indolyl, benzimidazolyl, naphthyl, quinolinyl,
quinoxalinyl, or isoquinolinyl ring. In certain embodiments,
R.sup.B is a substituted or unsubstituted phenyl ring.
[0159] The variable Y is hydrogen, --CN, or --NO.sub.2. In some
embodiments, Y is --NO.sub.2.
[0160] The variable R.sup.5a is hydrogen or a substituted or
unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl group.
In some embodiments, R.sup.5a is hydrogen or a substituted or
unsubstituted aliphatic group. In some other embodiments, R.sup.5a
is hydrogen or C.sub.1-6 aliphatic. In such embodiments, R.sup.5a
is selected from the group consisting of hydrogen, methyl, ethyl,
isopropyl and isobutyl. In certain such embodiments, R.sup.5a is
methyl.
[0161] The variable R.sup.5b is hydrogen or a substituted or
unsubstituted aliphatic, aryl, heteroaryl, or heterocyclyl group.
In some embodiments, R.sup.5b is hydrogen or a substituted or
unsubstituted aliphatic group. In some other embodiments, R.sup.5b
is hydrogen or C.sub.1-6 aliphatic. In such embodiments, R.sup.5b
is selected from the group consisting of hydrogen, methyl, ethyl,
isopropyl and isobutyl. In certain such embodiments, R.sup.5b is
hydrogen.
[0162] 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.
[0163] The variable A is 0, 1, or 2. In some embodiments, A is 0 or
1. In certain embodiments, A is 0.
[0164] The variable n is 0, or 1. In certain embodiments, n is 0.
In other certain embodiments, n is 1.
[0165] In some embodiments, A is 0; R.sup.a is hydrogen, C.sub.1-6
aliphatic, C.sub.1-6 fluoroaliphatic, or
--(CH.sub.2).sub.m--CH.sub.2--R.sup.B; R.sup.a1 is hydrogen,
C.sub.1-6 aliphatic, C.sub.1-6 fluoroaliphatic,
--(CH.sub.2).sub.m--CH.sub.2--R.sup.B, or
--(CH.sub.2).sub.m--CH(R.sup.5a)--OR.sup.5b; P is R.sup.c--C(O)--
or R.sup.c--S(O).sub.2--; R.sup.c is --R.sup.D; and m is 0 or
1.
[0166] In some other embodiments, A is 0; R.sup.a is C.sub.1-6
aliphatic or --(CH.sub.2).sub.m--CH.sub.2--R.sup.B; R.sup.a1 is
hydrogen, --(CH.sub.2).sub.m--CH.sub.2--R.sup.B, or
--(CH.sub.2).sub.m--CH(R.sup.5a)--OR.sup.5b; P is R.sup.c--C(O)--
or R.sup.c--S(O).sub.2--; R.sup.c is --R.sup.D; and m is 0 or
1.
[0167] In some other embodiments, A is 0; R.sup.a is C.sub.1-6
aliphatic; R.sup.a1 is hydrogen,
--(CH.sub.2).sub.m--CH.sub.2--R.sup.B, or
--(CH.sub.2).sub.m--CH(R.sup.5a)--OR.sup.5b; P is R.sup.c--C(O);
R.sup.c is --R.sup.D; and m is 0 or 1.
[0168] In some other embodiments, A is 0; R.sup.a is isobutyl;
R.sup.a1 is hydrogen, C.sub.1-6 aliphatic, C.sub.1-6
fluoroaliphatic, --(CH.sub.2).sub.m--CH.sub.2--R.sup.B, or
--(CH.sub.2).sub.m--CH(R.sup.5a)--OR.sup.5b; P is R.sup.c--C(O)--;
R.sup.c is --R.sup.D; and m is 0 or 1.
[0169] In yet some other embodiments, A is 0; R.sup.a is isobutyl;
R.sup.a1 is hydrogen, C.sub.1-6 aliphatic,
--(CH.sub.2).sub.m--CH.sub.2--R.sup.B, or
--(CH.sub.2).sub.m--CH(R.sup.5a)--OR.sup.5b; P is R.sup.c--C(O)--;
R.sup.c is --R.sup.D; and m is 0 or 1.
[0170] In still yet some other embodiments, A is 0; R.sup.a is
isobutyl; R.sup.a1 is hydrogen,
--(CH.sub.2).sub.m--CH.sub.2--R.sup.B, or
--(CH.sub.2).sub.m--CH(R.sup.5a)--OR.sup.5b; P is R.sup.c--C(O)--;
R.sup.c is --R.sup.D; and m is 0 or 1.
[0171] In certain embodiments, A is 0; R.sup.a is isobutyl;
R.sup.a1 is --CH.sub.2--R.sup.B, and R.sup.B is phenyl; P is
R.sup.c--C(O)--; R.sup.c is --R.sup.D; and R.sup.D is
2-pyrazinyl.
[0172] In other certain embodiments, A is 0; R.sup.a is isobutyl;
R.sup.a1 is hydrogen; P is R.sup.c--C(O)--; R.sup.c is --R.sup.D;
and R.sup.D is 2,5-dichlorophenyl.
[0173] In yet other certain embodiments, A is 0; R.sup.a is
isobutyl; R.sup.a1 is --CH(R.sup.5a)--OR.sup.5b; R.sup.5a is
C.sub.1-6 aliphatic; R.sup.5b is hydrogen; P is R.sup.c--C(O)--; W
is --R.sup.D; and R.sup.D is 6-phenyl-2-pyridinyl-.
[0174] In some embodiments, the compound of formula (I) is
characterized by formula (I-1):
##STR00009##
or a crystalline form thereof.
[0175] In some other embodiments, the compound of formula (I) is
characterized by formula (I-15):
##STR00010##
or a crystalline form thereof.
[0176] In yet some other embodiments, the compound of formula (I)
is characterized by formula (I-18):
##STR00011##
or a crystalline form thereof.
General Synthetic Methodology
[0177] The compounds of formula (I) can be prepared by
esterification of corresponding boronic acids. Such boronic acid
compounds can be prepared from methods known to one of ordinary
skill in the art. See, e.g., Adams et. al., U.S. Pat. No.
5,780,454; Pickersgill et al., International Patent Publication WO
2005/097809. An exemplary synthetic route is set forth in Scheme 1
below.
##STR00012##
[0178] 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,
without limitation, acyl protecting groups, e.g., formyl, acetyl
(Ac), succinyl (Suc), and methoxysuccinyl; and urethane protecting
groups, e.g., tert-butoxycarbonyl (Boc), benzyloxycarbonyl (Cbz),
and fluorenylmethoxycarbonyl (Fmoc). Optionally, PG is hydrogen and
deprotection is not necessary. The peptide coupling reaction can be
conducted by prior conversion of the carboxylic acid moiety of
compound ii to an activated ester or acid halide, e.g., an
O--(N-hydroxysuccinnimide) 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,
without limitation, carbodiimide reagents, e.g.,
dicyclohexylcarbodiimide (DCC) or
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC); phosphonium
reagents, e.g.,
(benzotriazol-1-yloxy)tris(dimethylamino)phosphonium
hexafluorophosphate (BOP); and uronium reagents, e.g.,
O-(1H-benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
tetrafluoroborate (TBTU).
[0179] Compound iii is then coupled with an amino-group blocking
moiety, to afford compound iv. The peptide coupling conditions
described above for the coupling of compounds i and ii are also
suitable for coupling compound iii with the amino-group blocking
moiety. Deprotection of the boronic acid moiety then affords
compound v. The deprotection step preferably is accomplished by
transesterification in a biphasic mixture comprising the boronic
ester compound iv, an organic boronic acid acceptor, a lower
alkanol, a C.sub.5-8 hydrocarbon solvent, and aqueous mineral acid.
Other reagents that can be used for deprotection of the boronic
acid moiety include, without limitation, BCl.sub.3, lithium
aluminium hydride and NaIO.sub.4.
##STR00013##
[0180] Alternatively, the order of coupling reactions can be
reversed, as shown in Scheme 2. Thus, an O-protected amino acid vi
is first coupled with an amino-group blocking moiety, followed by
ester hydrolysis, to form compound vii. Optionally, PG' is H and
ester hydrolysis is not needed, leading directly to compound vii.
Coupling with compound i and boronic acid deprotection are then
accomplished as described above for Scheme 1 to afford compound
v.
[0181] Compound v is reacted with the appropriate alpha-hydroxy
carboxylic acid or beta-hydroxy carboxylic acid to afford the
compound of formula (I) as shown in Scheme 3.
##STR00014##
[0182] The conversion of v to the compound of formula (I) can be
accomplished under esterification conditions employing
approximately a molar equivalent of the alpha-hydroxy carboxylic
acid or beta-hydroxy carboxylic acid in a solvent such as ethyl
acetate at a temperature of between about 40.degree. C. and about
80.degree. C. The conversion of v to the compound of formula (I)
can also be accomplished as described above employing a molar
excess of the alpha-hydroxy carboxylic acid or beta-hydroxy
carboxylic acid. Examples of other suitable solvents for this
conversion include, but are not limited to, methyl isobutyl ketone,
acetone, acetonitrile, 2-methyltetrahydrofuran, anisole, isopropyl
acetate, dimethoxyethane, tetrahydrofuran, dioxane,
dichloromethane, toluene, heptane, methyl-cyclohexane,
tert-butylmethyl ether, and mixtures thereof The choice of the
solvent will depend partly on the solubility of the alpha-hydroxy
carboxylic acid or beta-hydroxy carboxylic acid used. The
temperature selected for the conversion of v to the compound of
formula (I) will depend partly on the boiling point of the solvent
or solvent mixture used.
[0183] The conversion of v to the compound of formula (I) may 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-diazabicycle[4.3.0]non-5-ene,
N,N'diisopropylethylamine, or a mixture thereof.
[0184] The compound of formula v and the alpha-hydroxy carboxylic
acid or beta-hydroxy carboxylic acid are heated together in the
solvent of choice for a period of time. Following this period of
time, 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 may be uncontrolled or may be
controlled by the use of a cooling apparatus. The reaction mixture
may be stirred during this cooling period. Alternatively, the
compound of formula (I) can also be isolated from the reaction
mixture by cooling followed by evaporation of the solvent. The
reaction mixture may be seeded with crystals of the compound of
formula (I) in order to effect precipitation.
[0185] A co-solvent such as, but not limited to, heptane,
methylcyclohexane, toluene, tert-butylmethyl ether, ethyl acetate,
or a mixture thereof, may be added during the cooling period.
Following the addition of the co-solvent, the reaction mixture can
be cooled further leading to the precipitation of the compound of
formula (I). Alternatively, once the co-solvent is added, the
reaction mixture can then be heated again to generate a homogenous
solution, which is then cooled leading to the precipitation of the
compound of formula (I). The reaction mixture may be seeded with
crystals of the compound of formula (I) in order to effect
precipitation.
[0186] In other embodiments, the compound of formula (I) is
isolated in substantially pure form. In such embodiments, the
purity is about 80%, about 85%, about 90%, about 91%, about 92%,
about 93%, about 94%, about 95%, about 96%, about 97%, about 98%,
about 99%, or about 99.5%.
[0187] 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.
[0188] The compound of formula (I) can also be generated by the
co-lyophilization of compound v and the alpha-hydroxy carboxylic
acid or beta-hydroxy carboxylic acid. This is accomplished by
subjecting an aqueous solution comprising the compound of formula v
and a molar excess of the alpha-hydroxy carboxylic acid or
beta-hydroxy carboxylic acid to a lyophilization procedure. In some
embodiments, the aqueous solution additionally comprises a
water-miscible co-solvent. Examples of suitable co-solvents
include, but are not limited to, tert-butyl alcohol, methanol,
ethanol, and mixtures thereof The co-lyophilization results in a
composition that contains the compound of formula (I) and the
excess alpha-hydroxy carboxylic acid or beta-hydroxy carboxylic
acid.
Uses, Formulation, and Administration
[0189] The present invention provides compounds that are potent
inhibitors of the proteasome. The compounds can be assayed in vitro
or in vivo for their ability to inhibit proteasome-mediated peptide
hydrolysis or protein degradation.
[0190] In another aspect, therefore, the invention provides a
method for inhibiting one or more peptidase activities of a
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
acid anhydride thereof.
[0191] The invention also provides a method for inhibiting cell
proliferation, comprising contacting a cell in which such
inhibition is desired with a compound described herein. The phrase
"inhibiting cell proliferation" is used to denote the ability of a
compound of the invention to inhibit cell number or cell growth in
contacted cells as compared to cells not contacted with the
inhibitor. An assessment of cell proliferation can be made by
counting cells using a cell counter or by an assay of cell
viability, e.g., an MTT or WST assay. Where the cells are in a
solid growth (e.g., a solid tumor or organ), such an assessment of
cell proliferation can be made by measuring the growth, e.g., with
calipers, and comparing the size of the growth of contacted cells
with non-contacted cells.
[0192] Preferably, the growth of cells contacted with the inhibitor
is retarded by at least about 50% as compared to growth of
non-contacted cells. In various embodiments, cell proliferation of
contacted cells is inhibited by at least about 75%, at least about
90%, or at least about 95% as compared to non-contacted cells. In
some embodiments, the phrase "inhibiting cell proliferation"
includes a reduction in the number of contacted cells, as compared
to non-contacted cells. Thus, a proteasome inhibitor that inhibits
cell proliferation in a contacted cell may induce the contacted
cell to undergo growth retardation, to undergo growth arrest, to
undergo programmed cell death (i.e., apoptosis), or to undergo
necrotic cell death.
[0193] In another aspect, the invention provides a pharmaceutical
composition comprising a compound of formula (I), or a
pharmaceutically acceptable salt thereof, and a pharmaceutically
acceptable carrier.
[0194] In some embodiments, the composition also comprises the free
alpha-hydroxy carboxylic acid or a salt thereof or the beta-hydroxy
carboxylic acid or a salt thereof. In such embodiments, the
alpha-hydroxy carboxylic acid or a salt thereof or beta-hydroxy
carboxylic acid or a salt thereof 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 alpha hydroxy carboxylic acid or a salt
thereof or beta-hydroxy carboxylic acid or a 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.
[0195] If a pharmaceutically acceptable salt of the compound of the
invention is utilized in these compositions, the salt preferably is
derived from an inorganic or organic acid or base. For reviews of
suitable salts, see, e.g., 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.
[0196] Nonlimiting examples of suitable acid addition salts include
the following: acetate, adipate, alginate, aspartate, benzoate,
benzene sulfonate, bisulfate, butyrate, citrate, camphorate,
camphor sulfonate, cyclopentanepropionate, digluconate,
dodecylsulfate, 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 and undecanoate.
[0197] Suitable base addition salts include, without limitation,
ammonium salts, alkali metal salts, such as lithium, sodium and
potassium salts; alkaline earth metal salts, such as calcium and
magnesium salts; other multivalent metal salts, such as zinc salts;
salts with organic bases, such as dicyclohexylamine,
N-methyl-D-glucamine, t-butylamine, ethylene diamine, ethanolamine,
and choline; and salts with amino acids such as arginine, lysine,
and so forth.
[0198] The term "pharmaceutically acceptable carrier" is used
herein to refer to a material that is compatible with a recipient
subject, preferably a mammal, more preferably a human, and is
suitable for delivering an active agent to the target site without
terminating the activity of the agent. The toxicity or adverse
effects, if any, associated with the carrier preferably are
commensurate with a reasonable risk/benefit ratio for the intended
use of the active agent.
[0199] The terms "carrier", "excipient" or "vehicle" are used
interchangeably herein, and include any and all solvents, diluents,
and other liquid vehicles, dispersion or suspension aids, surface
active agents, pH modifiers, isotonic agents, thickening or
emulsifying agents, preservatives, solid binders, lubricants and
the like, as suited to the particular dosage form desired.
Remington: The Science and Practice of Pharmacy, 20th Ed., ed. A.
Gennaro, Lippincott Williams & Wilkins, 2000 discloses various
carriers used in formulating pharmaceutically acceptable
compositions and known techniques for the preparation thereof.
Strickley, Pharmaceutical Research, 21(2) 201-230 (2004) reviews
pharmaceutically acceptable excipients 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 invention, such as by producing any
undesirable biological effect or otherwise interacting in a
deleterious manner with any other component(s) of the
pharmaceutically acceptable composition, its use is contemplated to
be within the scope of this invention. Some examples of materials
which can serve as pharmaceutically acceptable carriers include,
but are not limited to, ion exchangers, alumina, aluminum stearate,
lecithin, serum proteins, such as human serum albumin, buffer
substances such as phosphates, carbonates, magnesium hydroxide and
aluminum hydroxide, glycine, sorbic acid, or potassium sorbate,
partial glyceride mixtures of saturated vegetable fatty acids,
water, pyrogen-free water, salts or electrolytes such as protamine
sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate,
sodium chloride, and zinc salts, colloidal silica, magnesium
trisilicate, polyvinyl pyrrolidone, polyacrylates, waxes,
polyethylene-polyoxypropylene-block polymers, wool fat, sugars such
as lactose, glucose, sucrose, and mannitol, starches such as corn
starch and potato starch, cellulose and its derivatives such as
sodium carboxymethyl cellulose, ethyl cellulose and cellulose
acetate, powdered tragacanth; malt, gelatin, talc, excipients such
as cocoa butter and suppository waxes, oils such as peanut oil,
cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and
soybean oil, glycols such as propylene glycol and polyethylene
glycol, esters such as ethyl oleate and ethyl laurate, agar,
alginic acid, isotonic saline, Ringer's solution, alcohols such as
ethanol, isopropyl alcohol, hexadecyl alcohol, and glycerol,
cyclodextrins such as hydroxypropyl .beta.-cyclodextrin and
sulfobutylether .beta.-cyclodextrin, lubricants such as sodium
lauryl sulfate and magnesium stearate, petroleum hydrocarbons such
as mineral oil and petrolatum. Coloring agents, releasing agents,
coating agents, sweetening, flavoring and perfuming agents,
preservatives and antioxidants can also be present in the
composition, according to the judgment of the formulator.
[0200] The pharmaceutical compositions of the invention can be
manufactured by methods well known in the art such as conventional
granulating, mixing, dissolving, encapsulating, lyophilizing, or
emulsifying processes, among others. Compositions may be produced
in various forms, including granules, precipitates, or
particulates, powders, including freeze dried, rotary dried or
spray dried powders, amorphous powders, tablets, capsules, syrup,
suppositories, injections, emulsions, elixirs, suspensions or
solutions.
[0201] According to a preferred embodiment, the compositions of
this invention are formulated for pharmaceutical administration to
a mammal, preferably a human being. Such pharmaceutical
compositions of the present invention may be administered orally,
parenterally, by inhalation spray, topically, rectally, nasally,
buccally, vaginally or via an implanted reservoir. The term
"parenteral" as used herein includes subcutaneous, intravenous,
intramuscular, intra-articular, intra-synovial, intrasternal,
intrathecal, intrahepatic, intralesional and intracranial injection
or infusion techniques. Preferably, the compositions are
administered orally, intravenously, or subcutaneously. The
formulations of the invention may be designed to be short-acting,
fast-releasing, or long-acting. Still further, compounds can be
administered in a local rather than systemic means, such as
administration (e.g., by injection) at a tumor site.
[0202] Liquid dosage forms for oral administration include, but are
not limited to, pharmaceutically acceptable emulsions,
microemulsions, solutions, suspensions, syrups and elixirs. In
addition to the active compounds, the liquid dosage forms may
contain inert diluents commonly used in the art such as, for
example, water or other solvents, solubilizing agents and
emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl
carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate,
propylene glycol, 1,3-butylene glycol, cyclodextrins,
dimethylformamide, oils (in particular, cottonseed, groundnut,
corn, germ, olive, castor, and sesame oils), glycerol,
tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid
esters of sorbitan, and mixtures thereof. Besides inert diluents,
the oral compositions can also include adjuvants such as wetting
agents, emulsifying and suspending agents, sweetening, flavoring,
and perfuming agents.
[0203] Injectable preparations, for example, sterile injectable
aqueous or oleaginous suspensions may be formulated according to
the known art using suitable dispersing or wetting agents and
suspending agents. The sterile injectable preparation may also be a
sterile injectable solution, suspension or emulsion in a
parenterally acceptable diluent or solvent, for example, as a
solution in 1,3-butanediol. Among the acceptable vehicles and
solvents that may be employed are water, Ringer's solution, U.S.P.
and isotonic sodium chloride solution. In addition, sterile, fixed
oils are conventionally employed as a solvent or suspending medium.
For this purpose any bland fixed oil can be employed including
synthetic mono- or diglycerides. In addition, fatty acids such as
oleic acid are used in the preparation of injectables. The
injectable formulations can be sterilized, for example, by
filtration through a bacterial-retaining filter, or by
incorporating sterilizing agents in the form of sterile solid
compositions which can be dissolved or dispersed in sterile water
or other sterile injectable medium prior to use. Compositions
formulated for parenteral administration may be injected by bolus
injection or by timed push, or may be administered by continuous
infusion.
[0204] Solid dosage forms for oral administration include capsules,
tablets, pills, powders, and granules. In such solid dosage forms,
the active compound is mixed with at least one inert,
pharmaceutically acceptable excipient or carrier such as sodium
citrate or dicalcium phosphate and/or a) fillers or extenders such
as starches, lactose, cellulose, sucrose, glucose, mannitol, and
silicic acid, b) binders such as, for example,
carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,
sucrose, and acacia, c) humectants such as glycerol, d)
disintegrating agents such as agar-agar, calcium carbonate, potato
or tapioca starch, alginic acid, certain silicates, crospovidone,
cellulose, croscaramellose sodium, sodium starch glycolate, and
sodium carbonate, e) solution retarding agents such as paraffin, f)
absorption accelerators such as quaternary ammonium compounds, g)
wetting agents such as, for example, cetyl alcohol and glycerol
monostearate, h) absorbents such as kaolin and bentonite clay, and
i) lubricants such as talc, calcium stearate, magnesium stearate,
sodium stearyl fumarate, stearic acid, solid polyethylene glycols,
sodium lauryl sulfate, glyceryl behenate, and mixtures thereof In
the case of capsules, tablets and pills, the dosage form may also
comprise buffering agents such as phosphates or carbonates.
[0205] Solid compositions of a similar type may also be employed as
fillers in soft and hard-filled gelatin capsules using such
excipients as lactose or milk sugar as well as high molecular
weight polyethylene glycols and the like. The solid dosage forms of
tablets, dragees, capsules, pills, and granules can be prepared
with coatings and shells such as enteric coatings and other
coatings well known in the pharmaceutical formulating art. They may
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 may also be employed as fillers in
soft and hard-filled gelatin capsules using such excipients as
lactose or milk sugar as well as high molecular weight polethylene
glycols and the like.
[0206] The active compounds can also be in micro-encapsulated form
with one or more excipients as noted above. The solid dosage forms
of tablets, dragees, capsules, pills, and granules can be prepared
with coatings and shells such as enteric coatings, release
controlling coatings and other coatings well known in the
pharmaceutical formulating art. In such solid dosage forms the
active compound may be admixed with at least one inert diluent such
as sucrose, lactose or starch. Such dosage forms may also comprise,
as is normal practice, additional substances other than inert
diluents, e.g., tableting lubricants and other tableting aids such
as magnesium stearate and microcrystalline cellulose. In the case
of capsules, tablets and pills, the dosage forms may also comprise
buffering agents. They may optionally contain opacifying agents and
can also be of a composition that they release the active
ingredient(s) only, or preferentially, in a certain part of the
intestinal tract, optionally, in a delayed manner. Examples of
embedding compositions that can be used include polymeric
substances and waxes. In some embodiments, the excipients or
carriers may include, but are not limited to sodium stearyl
fumarate, carboxymethylcellulose, magnesium stearate, crospovidone,
ethylcellulose, talc, and silicified microcrystalline
cellulose.
[0207] Dosage forms for topical or transdermal administration of a
compound of this invention include ointments, pastes, creams,
lotions, gels, powders, solutions, sprays, inhalants or patches.
The active component is admixed under sterile conditions with a
pharmaceutically acceptable carrier and any needed preservatives or
buffers as may be required. Ophthalmic formulation, ear drops, and
eye drops are also contemplated as being within the scope of this
invention. Additionally, the present invention contemplates the use
of transdermal patches, which have the added advantage of providing
controlled delivery of a compound to the body. Such dosage forms
can be made by dissolving or dispensing the compound in the proper
medium. Absorption enhancers can also be used to increase the flux
of the compound across the skin. The rate can be controlled by
either providing a rate controlling membrane or by dispersing the
compound in a polymer matrix or gel.
[0208] In some embodiments, the present invention provides
pharmaceutical compositions comprising the compound of formula (I),
and additional excipients described herein. In some other
embodiments, the present invention provides pharmaceutical
compositions comprising the compound of formula (II), and
additional excipients described herein. In yet some other
embodiments, the present invention provides pharmaceutical
compositions comprising the compound of formula (III) or (IV), and
additional excipients described herein.
[0209] In further embodiments, the present invention provides
pharmaceutical compositions comprising the citrate ester of
compound (VIII-1), and additional excipients described herein. In
other further embodiments, the present invention provides
pharmaceutical compositions comprising the citrate ester of
compound (VIII-15), and additional excipients described herein. In
further embodiments, the present invention provides pharmaceutical
compositions comprising the citrate ester of (VIII-18) and
additional excipients described herein.
##STR00015##
[0210] In further embodiments, the present invention provides
pharmaceutical compositions comprising the compound (I-1), or a
crystalline form thereof. In still yet further embodiments, the
present invention provides pharmaceutical compositions comprising
the compound (I-15), or a crystalline form thereof In still yet
some further embodiments, the present invention provides
pharmaceutical compositions comprising the compound (I-18), or a
crystalline form thereof.
[0211] The following description of pharmaceutical compositions and
methods for preparation of said pharmaceutical compositions are
applicable to the compounds of formulas (I), (II), (III), (IIIa),
(IV), or (IVa) and various embodiments of these formulas as
described herein. The following description of pharmaceutical
compositions and methods for preparation of said pharmaceutical
compositions are also applicable to the compounds (I-1), (I-15), or
(I-18).
[0212] In one embodiment, the pharmaceutical composition comprises
the 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% of a crystalline form. In yet
another embodiment, the compound of formula (I) in the
pharmaceutical composition is a crystalline form.
[0213] In some embodiments, the pharmaceutical formulations of the
invention provide stable solid oral dosage forms of active
compound, by using excipients having low water or low-moisture
content, and manufactured using dry or non-aqueous formulation
processes.
[0214] In one embodiment, the pharmaceutical composition is an oral
pharmaceutical dosage form, selected from the group consisting of
capsules, tablets, pills, powders, and granules. In another
embodiment, the oral pharmaceutical dosage form is a capsule,
wherein the capsule is a polymer-based capsule selected from the
group consisting of gelatin, hydroxypropylmethyl cellulose (HPMC),
fish gelatin, and pullulan. In yet another embodiment, the
polymer-based capsule is selected from the group consisting of
gelatin, and hydroxypropylmethyl cellulose. In still yet another
embodiment, the polymer-based capsule is a hard gelatin
capsule.
[0215] 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.
[0216] In another embodiment, the pharmaceutical composition
further comprises an optional flow-aid, and an optional buffer. In
yet another embodiment, the pharmaceutical composition comprises
about 0.2% to about 3% of the compound of formula (I), or a
crystalline form thereof, about 86.5% to about 99.8% of a filler,
optionally up to about 1.5% of a lubricant, optionally up to about
5% of a flow-aid, and optionally up to about 5% of a buffer, by
weight as a percentage of total weight.
[0217] In another embodiment, the pharmaceutical composition
comprises about 0.2% to about 12% of the compound of formula (I),
or a crystalline form thereof, about 76.5% to about 99.8% of a
filler, optionally up to about 1.5% of a lubricant, optionally up
to about 5% of a flow-aid, and optionally up to about 5% of a
buffer, by weight as a percentage of total weight.
[0218] 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. In 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.
[0219] Suitable fillers include, but are not limited to, powdered
cellulose, microcrystalline cellulose, silicified microcrystalline
cellulose, high density microcrystalline cellulose, low-moisture
microcrystalline cellulose, pregeletanized 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, pregeletanized
starch, and mixtures thereof.
[0220] 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 from about 98% to about 99.75%, by weight as a percentage of
total weight. In yet some other embodiments, when a lubricant is
present, the amount of filler is reduced by the corresponding
percent amount of 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.
[0221] In some embodiments, the filler comprises a first filler and
a second filler. The first filler is present in an amount of 0% to
about 99.8%, by weight as a percentage of total weight, and the
second filler is present in an amount of 0% to about 99.8% by
weight as a percentage of total weight, as long as the total amount
of filler is no greater than about 99.8%. 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, as long as the total amount of filler
is not greater than about 99.8%, by weight as a percentage of total
weight.
[0222] In some embodiments, the first filler is selected from the
group consisting of low-moisture microcrystalline cellulose, sodium
starch glycolate, pregeletanized starch, and mixtures thereof. In
some embodiments, the second filler is selected from the group
consisting of low-moisture microcrystalline cellulose, sodium
starch glycolate, pregeletanized starch, and mixtures thereof
[0223] Suitable lubricants include, but are not limited to,
magnesium stearate, glyceryl behenate, hydrogenated vegetable oil,
talc, zinc stearate, calcium stearate, sucrose stearate, sodium
stearyl fumarate, and mixtures thereof. In some embodiments, the
lubricant is magnesium stearate. In other embodiments, the
lubricant is present in an amount of up to about 1.5%, by weight as
a percentage of total weight. In yet some other embodiments, the
lubricant is present in an amount of about 1%, by weight as a
percentage of total weight.
[0224] Suitable flow-aids include, but are not limited to silicion
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 up to about 5%, by weight as a percentage of total
weight. In some other embodiments, the flow-aid is present in an
amount of about 1%, by weight as a percentage of total weight. In
yet some other embodiments, the flow-aid is present in an amount of
about 2%, by weight as a percentage of total weight.
[0225] Suitable buffers include sodium citrate, citric acid, and
mixtures thereof. In some embodiments, the buffer is sodium
citrate. In some other embodiments, the buffer is present in an
amount of up to about 5%, by weight as a percentage of total
weight. In yet some other embodiments, the buffer is present in an
amount of about 2%, by weight as a percentage of total weight.
[0226] In some embodiments, the pharmaceutical composition
comprises the compound of formula (I), or a crystalline form
thereof, a filler, and optionally a lubricant; wherein: [0227] the
alpha-hydroxy carboxylic acid or beta-hydroxy carboxylic acid is
citric acid; [0228] A is 0; [0229] R.sup.a is isobutyl; [0230]
R.sup.a1 is hydrogen, C.sub.1-6 aliphatic,
--(CH.sub.2).sub.m--CH.sub.2--R.sup.B, or
--(CH.sub.2).sub.m--CH(R.sup.5a)--OR.sup.5b; [0231] P is
R.sup.c--C(O)--; [0232] R.sup.c is --R.sup.D; [0233] m is 0 or 1;
[0234] the filler is selected from the group consisting of
low-moisture microcrystalline cellulose, sodium starch glycolate,
pregeletanized starch, and mixtures thereof; and [0235] the
lubricant, when present, is magnesium stearate.
[0236] In some embodiments, the pharmaceutical composition
comprises the compound of formula (I), or a crystalline form
thereof, a filler, and optionally a lubricant; wherein: [0237] the
compound of formula (I) is (I-1), (I-15) or (I-18); [0238] the
filler is selected from the group consisting of low-moisture
microcrystalline cellulose, sodium starch glycolate, pregeletanized
starch, and mixtures thereof; and [0239] the lubricant, when
present, is magnesium stearate.
[0240] In some embodiments, the pharmaceutical composition
comprises about 0.25% to about 2% of the compound of formula (I),
or a crystalline form thereof; and about 98% to about 99.75% of a
filler; wherein: [0241] the compound of formula (I) is (I-1),
(I-15) or (I-18); and [0242] the filler is selected from the group
consisting of low-moisture microcrystalline cellulose, sodium
starch glycolate, pregeletanized starch, and mixtures thereof.
[0243] In some embodiments, the pharmaceutical composition
comprises the compound of formula (I), or a crystalline form
thereof, a filler, optionally a lubricant; optionally a flow-aid;
and optionally a buffer; wherein: [0244] the alpha-hydroxy
carboxylic acid or beta-hydroxy carboxylic acid is citric acid;
[0245] A is 0; [0246] R.sup.a is isobutyl; [0247] R.sup.a1 is
hydrogen, C1-6 aliphatic, --(CH.sub.2).sub.m--CH.sub.2--R.sup.B, or
--(CH.sub.2).sub.m--CH(R.sup.5a)-OR.sup.5b; [0248] P is
R.sup.c--C(O)--; [0249] R.sup.c is --R.sup.D; [0250] m is 0 or 1;
[0251] the filler is selected from the group consisting of
low-moisture microcrystalline cellulose, sodium starch glycolate,
pregeletanized starch, and mixtures thereof; [0252] the lubricant,
when present, is magnesium stearate; [0253] the flow-aid, when
present, is talc; and [0254] the buffer, when present, is sodium
citrate.
[0255] In some embodiments, the pharmaceutical composition
comprises the compound of formula (I), or a crystalline form
thereof, a filler, optionally a lubricant; optionally a flow-aid;
and optionally a buffer, wherein: [0256] the compound of formula
(I) is (I-1), (I-15) or (I-18); [0257] the filler is selected from
the group consisting of low-moisture microcrystalline cellulose,
sodium starch glycolate, pregeletanized starch, and mixtures
thereof; [0258] the lubricant, when present, is magnesium stearate;
[0259] the flow-aid, when present, is talc; and [0260] the buffer,
when present, is sodium citrate.
[0261] In some embodiments, the pharmaceutical composition
comprises about 0.2% to about 3% of the compound of formula (I), or
a crystalline form thereof, about 86.5% to about 99.8% of a filler,
optionally up to about 1.5% of a lubricant, optionally up to about
5% of a flow-aid, and optionally up to about 5% of a buffer, by
weight as a percentage of total weight, wherein: [0262] the
compound of formula (I) is (I-1), (I-15) or (I-18); [0263] the
filler is selected from the group consisting of low-moisture
microcrystalline cellulose, sodium starch glycolate, pregeletanized
starch, and mixtures thereof; [0264] the lubricant, when present,
is magnesium stearate; [0265] the flow-aid, when present, is talc;
and [0266] the buffer, when present, is sodium citrate.
[0267] In some embodiments, the pharmaceutical composition
comprises the 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, the
pharmaceutical composition comprises the 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, pregeletanized
starch, and mixtures thereof; and the lubricant, when present, is
magnesium stearate.
[0268] In some embodiments, the pharmaceutical composition
comprises the 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.
[0269] In some embodiments, the pharmaceutical composition
comprises the compound of formula (I-1) Form 2, and low-moisture
microcrystalline cellulose. 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.
[0270] 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 pregeletanized starch. In yet some other
embodiments, the pharmaceutical composition comprises the compound
of formula (I-1) Form 2, pregeletanized 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 other further embodiments, the
pharmaceutical composition comprises the compound of formula (I-1)
Form 2, low-moisture microcrystalline cellulose, talc, magnesium
stearate, and pregeletanized 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.
[0271] When the compound of formula (I) is subjected to hydrolytic
conditions, the ester portion of the molecule is hydrolyzed to give
the compound of formula (VIII) in a 1:1 molecular ratio.
##STR00016##
[0272] Using an analytical method that involves hydrolytic
conditions for sample preparation, the amount of the compound of
formula (VIII) present in a test sample is measured (see e.g.
Analytical Test Method 1, below), by comparison to a reference
standard of known purity. Using an analytical method that does not
subject the sample to hydrolytic conditions, the amount of the
compound of formula (VIII) present in the sample of the compound of
formula (I) is measured by comparison to a reference standard of
known purity (see e.g. Analytical Test Method 2 below). Therefore,
the amount of the compound of formula (VIII) measured in Analytical
Test Method 1 minus the amount of the compound of formula (VIII)
measured in Analytical Test Method 2, gives the amount of the
compound of formula (VIII) in the sample that is derived from
hydrolysis of the compound of formula (I). Based on a 1:1 molecular
ratio for the conversion of the compound of formula (I) to the
compound of formula (VIII), a molecular weight conversion gives the
amount of compound of formula (I) present in the test sample.
[0273] It will be recognized that such analytical methods as
described directly above, and in the Experimental section below are
applicable in a similar manner to any of the compounds of formulas
(I), (II), (III), (IIIa), (IV), or (IVa) and various embodiments of
these formula as described herein. Such analytical methods as
described directly above and in the Experimental section below are
applicable in a similar manner to the compounds (I-1), (I-15), or
(I-18).
[0274] In some embodiments, the amount of the compound of formula
(VIII) present in a pharmaceutical composition is determined by
measuring the amount of the compound of formula (VIII) that is
present after subjecting to the sample to conditions under which
the compound of formula (I) is hydrolyzed to the compound of
formula (VIII).
[0275] In some embodiments, the amount of the compound of formula
(I-1), or a crystalline form thereof, present in a pharmaceutical
composition is expressed as the equivalent amount on a molar weight
basis of the compound of formula (VIII-1).
[0276] In some embodiments, the invention relates to a unit dose
pharmaceutical composition comprising the compound of formula
(I-1), or a crystalline form thereof.
[0277] In some other embodiments, the unit dose pharmaceutical
composition comprises the compound of formula (I-1), or a
crystalline form thereof, wherein the compound of formula (I-1) is
present in an amount equivalent to a molar weight basis of about
0.1 mg to about 3.0 mg of the compound of formula (VIII-1). In yet
some other embodiments, the unit dose pharmaceutical composition
comprises the compound of formula (I-1), or a crystalline form
thereof, wherein the compound of formula (I-1) is present in an
amount equivalent to a molar weight basis of about 0.15 mg to about
2.2 mg of the compound of formula (VIII-1). In still yet some other
embodiments, the unit dose pharmaceutical composition comprises the
compound of formula (I-1), or a crystalline form thereof, wherein
the compound of formula (I-1) is present in an amount equivalent to
a molar weight basis of about 0.18 mg to about 0.22 mg of the
compound of formula (VIII-1). In some further embodiments, the unit
dose pharmaceutical composition comprises the compound of formula
(I-1), or a crystalline form thereof, wherein the compound of
formula (I-1) is present in an amount equivalent to a molar weight
basis of about 0.46 mg to about 0.54 mg of the compound of formula
(VIII-1). In still some further embodiments, the unit dose
pharmaceutical composition comprises the compound of formula (I-1),
or a crystalline form thereof, wherein the compound of formula
(I-1) is present in an amount equivalent to a molar weight basis of
about 1.80 mg to about 2.20 mg of the compound of formula
(VIII-1).
[0278] In some embodiments, the amount of the compound of formula
(I-1), or a crystalline form thereof, present in a pharmaceutical
composition is expressed as the 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).
[0279] In some embodiments, the unit dose pharmaceutical
composition comprises about 0.143 mg to about 4.3 mg of the
compound of formula (I-1), or a crystalline form thereof, measured
as about 0.1 mg to about 3.0 mg of the compound of formula
(VIII-1), on a weight for weight basis.
[0280] In some other embodiments, the unit dose pharmaceutical
composition comprises about 0.214 mg to about 3.15 mg of the
compound of formula (I-1), or a crystalline form thereof, measured
as about 0.15 mg to about 2.2 mg of the compound of formula
(VIII-1), on a weight for weight basis.
[0281] In yet some other embodiments, the unit dose pharmaceutical
composition comprises about 0.258 mg to about 0.315 mg of the
compound of formula (I-1), or a crystalline form thereof, measured
as about 0.18 mg to about 0.22 mg of the compound of formula
(VIII-1), on a weight for weight basis.
[0282] In still yet some other embodiments, the unit dose
pharmaceutical composition comprises about 0.659 mg to about 0.773
mg of the compound of formula (I-1), or a crystalline form thereof,
measured as about 0.46 mg to about 0.54 mg of the compound of
formula (VIII-1), on a weight for weight basis.
[0283] In some further embodiments, the unit dose pharmaceutical
composition comprises about 2.58 mg to about 3.15 mg of the
compound of formula (I-1), or a crystalline form thereof, measured
as about 1.80 mg to about 2.20 mg of the compound of formula
(VIII-1), on a weight for weight basis.
[0284] In some embodiments, the invention provides a process for
the production of 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, comprising the steps
of: [0285] (a-1) mixing together screened filler and screened
compound of formula (I), or a crystalline form thereof, in a bag;
[0286] (a-2) passing the resulting mixture from step (a-1) through
a screen, then blending; [0287] (a-3) screening additional filler
through the same screen, passing it through the same bag, and
blending in the same blending apparatus; [0288] (a-4) repeating
step (a-3) up to two times; [0289] (a-5) taking the resulting
mixture from step (a-4), and encapsulating it using a
capsule-filling system; and [0290] (a-6) weight-sorting the
resulting capsules from step (a-5).
[0291] In some embodiments, step (a-3) may be repeated three or
more times.
[0292] When a lubricant is present in the pharmaceutical
composition, the invention provides a process for the production of
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, comprising the steps of: [0293] (b-1)
mixing together screened filler, and screened compound of formula
(I), or a crystalline form thereof, in a bag; [0294] (b-2) passing
the resulting mixture from step (b-1) through a screen, then
blending; [0295] (b-3) screening additional filler through the same
screen, passing it through the same bag, and blending in the same
blending apparatus; [0296] (b-4) repeating step (b-3) up to two
times; [0297] (b-5) blending together the mixture from step (b-4),
and screened lubricant; [0298] (b-6) taking the resulting mixture
from step (b-5), and encapsulating it using a capsule-filling
system; and [0299] (b-7) weight-sorting the resulting capsules from
step (b-6).
[0300] In some embodiments, step (b-3) may be repeated three or
more times. When additional components are present in the
pharmaceutical composition, such as buffer, second filler, or
flow-aid, they may be added in any of steps (b-1) or (b-3). The
total amount of each component of the pharmaceutical composition
may be added in one step or may be broken into several amounts,
which may or may not be of equal weight, and added in individual
occurrences of steps (b-1) or (b-3).
[0301] In some embodiments, the invention provides a process for
the production of 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, comprising the steps
of: [0302] (c-1) passing filler through a screen, then placing in a
high shear mixing apparatus; [0303] (c-2) passing the compound of
formula (I), or a crystalline form thereof, through a screen, then
placing in the same high shear mixing apparatus; [0304] (c-3)
passing filler through a screen, then placing in the same high
shear mixing apparatus; [0305] (c-4) mixing using the same high
shear mixing apparatus for less than 10 minutes; [0306] (c-5)
taking the resulting mixture from step (c-4), and encapsulating it
using a capsule-filling system; and [0307] (c-6) weight-sorting the
resulting capsules from step (c-5).
[0308] In some embodiments, when using the high shear mixing
apparatus, additional components that are present in the
pharmaceutical composition may be added by repeating either step
(c-1) or step (c-3).
[0309] In some embodiments, the compound of formula (I) used in the
processes for preparation of solid oral dosage forms 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 processes for preparation of solid oral dosage forms described
above is (I-1).
[0310] The process steps outlined above can take place using
conventional apparatus and equipment. For a review, see e.g.
Remington: The Science and Practice of Pharmacy, 21.sup.st Ed.,
Lippincott Williams & Wilkins, 2005.
[0311] The blending steps outlined above can take place in any
conventional blending apparatus. In some embodiments, the blending
time for each individual blending step is between about 1 minute
and about 45 minutes. In some other embodiments, the blending time
for each individual blending step is between about 1 minute and
about 20 minutes. In yet some other embodiments, the blending time
for each individual blending step is between about 2 minutes and
about 15 minutes.
[0312] The mixing step outlined above can take place in any
conventional polyethylene bag. In some embodiments, the mixing step
takes between about 30 seconds and 5 minutes. In some embodiments,
the mixing step outlined above can take place in a stainless steel
container.
[0313] The mixing step using the high shear mixing apparatus can
take place in any conventional high shear mixing apparatus. An
example of such a high shear mixing apparatus is sold as Lab High
Shear Granulator (Key International, Inc., Englishtown, N.J.). In
some embodiments, the mixing is performed for less than about 10
minutes. In some other embodiments, the mixing is performed for
less than about 5 minutes.
[0314] The capsule filling step outlined above can take place in
any conventional capsule filling system or apparatus. In some
embodiments, the capsule filling system is semi-automated, and can
handle 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 apparatus is sold as
ProFill 100 (Torpac, Inc., Fairfield, N.J., USA).
[0315] In some embodiments, the capsules are hard gelatin capsules,
sold as Coni-Snap.RTM. (Capsugel, Peapack, N.J.). One of skill in
the art will be able to select the appropriate capsule size and
color. In some embodiments, the capsules have a fill weight of 85
mg, 120 mg, or 150 mg.
[0316] The weight-sorting step outlined above can take place using
any conventional weight-sorting apparatus or machine. An example of
a weight-sorting apparatus or machine is sold as the SADE SP Bench
Top Tablet and Capsule Weight Sorter (AC Compacting LLC, North
Brunswick, N.J., USA).
[0317] In some embodiments, the capsules are packaged in bottles,
foil pouches or blister packs. In some other embodiments, the
capsules are packaged in heat-induction sealed high-density
polyethylene (HDPE) bottles. In another embodiment, the capsules
are packaged in air-tight sealed foil pouches. In yet another
embodiment, the capsules are packaged in foil-foil blister packs.
In some other embodiments, the capsules are packaged with a
desiccant.
[0318] The physical and chemical stability of the oral
pharmaceutical dosage form may be tested in a conventional manner,
for example, the measurement of dissolution, disintegration time,
assay for the compound of formula (I) degradation products, after
storage at different temperatures for different lengths of
time.
[0319] In some other embodiments, the invention provides
pharmaceutical compositions for parenteral use. In yet some other
embodiments, the invention provides liquid pharmaceutical
compositions for parenteral or oral use.
[0320] In some embodiments, the compound of formula (I) is
formulated as a lyophilized powder, in a manner analogous to that
described in Plamondon et al., WO 02/059131, hereby incorporated by
reference in its entirety. In such embodiments, an aqueous mixture
comprising a alpha-hydroxy carboxylic acid or a beta-hydroxy
carboxylic acid is lyophilized to form the compound of formula
(I).
[0321] In some embodiments, the lyophilized powder also comprises
free alpha-hydroxy carboxylic acid, or beta hydroxy carboxylic
acid. Preferably, the free alpha-hydroxy carboxylic acid or beta
hydroxy carboxylic 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 wherein the alpha-hydroxy carboxylic
acid or beta-hydroxy carboxylic acid compound is citric acid, the
lyophilized powder comprises free citric acid and the corresponding
boronate 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.
[0322] In some embodiments, the lyophilized powder comprises citric
acid and a compound of formula (I), substantially free of other
components. However, the composition can further comprise one or
more other pharmaceutically acceptable excipients, carriers,
diluents, fillers, salts, buffers, bulking agents, stabilizers,
solubilizers, and other materials well known in the art. The
preparation of pharmaceutically acceptable formulations containing
these materials is described in, e.g., Remington: The Science and
Practice of Pharmacy, 20th Ed., ed. A. Gennaro, Lippincott Williams
& Wilkins, 2000, or latest edition, and Strickley,
Pharmaceutical Research, 21(2) 201-230 (2004).
[0323] Upon dissolution in aqueous medium, equilibrium is
established between the boronate ester compound of formula (I) and
the corresponding free boronic acid compound. In some embodiments,
equilibrium is reached quickly, e.g., within 1-15 minutes, after
the addition of aqueous medium. The relative concentrations of
boronate ester, boronic acid, and any intermediate species present
at equilibrium is dependent upon parameters such as, e.g., the pH
of the solution, temperature, the nature of the alpha-hydroxy
carboxylic acid or beta-hydroxy carboxylic acid, and the ratio of
the alpha-hydroxy carboxylic acid or beta-hydroxy carboxylic acid
to boronate ester compound of formula (I) present in the
lyophilized powder.
[0324] In some embodiments, the pharmaceutical composition
comprises the compound of formula (I), a bulking agent, and a
buffer. In some other embodiments, the pharmaceutical composition
comprises the compound of formula (I), a bulking agent, and a
buffer in a lyophilized powder.
[0325] In some embodiments, the compound of formula (I) is
pre-formed. 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, the compound (I-1) is
pre-formed. In still yet some other embodiments, the compound
(I-15) is formed in situ from compound (VIII-15).
[0326] Suitable bulking agents include glycine. In some
embodiments, the amount of the bulking agent present is about 1%
weight/volume (w/v) to about 5% w/v. In some other embodiments, the
amount of the bulking agent present is about 3% w/v.
[0327] Suitable buffers include sodium citrate, citric acid, and
mixtures thereof. In some embodiments, the buffer is sodium
citrate, and citric acid.
[0328] In some embodiments, the buffer is present in a
concentration of about 45 mM to about 65 mM. In some other
embodiments, the buffer is present in a concentration of about 50
mM to about 60 mM.
[0329] In some embodiments, the ratio of the buffer to the compound
of formula (I) is from about 50:1 to about 10:1. In some other
embodiments, the ratio of the buffer to the compound of formula (I)
is from about 30:1 to about 10:1. In yet some other embodiments,
the ratio of the buffer to the compound of formula (I) is about
20:1.
[0330] In some embodiments, the pH of the pharmaceutical
composition is between about pH 4.7 and pH 6.1. The pH of the
pharmaceutical composition can be adjusted using any suitable
inorganic acid or organic acid.
[0331] In some embodiments, the pharmaceutical composition
comprises the compound of formula (I), a bulking agent, and a
buffer; wherein: [0332] the alpha-hydroxy carboxylic acid or
beta-hydroxy carboxylic acid is citric acid; [0333] A is 0; [0334]
R.sup.a is isobutyl; [0335] R.sup.a1 is hydrogen, C.sub.1-6
aliphatic, --(CH.sub.2).sub.m--CH.sub.2--R.sup.D, or
--(CH.sub.2).sub.m--CH(R.sup.5a)--OR.sup.5b; [0336] P is
R.sup.c--C(O)--; [0337] R.sup.c is --R.sup.D; [0338] m is 0 or 1;
[0339] the bulking agent is glycine; and [0340] the buffer is
sodium citrate, and citric acid.
[0341] In some embodiments, the pharmaceutical composition
comprises the compound of formula (I), a bulking agent, and a
buffer; wherein: [0342] the compound of formula (I) is represented
by compounds (I-1), (I-15) or (I-18); [0343] the bulking agent is
glycine; and [0344] the buffer is sodium citrate, and citric
acid.
[0345] In some embodiments, the pharmaceutical composition
comprises the compound of formula (I), a bulking agent, and a
buffer in a lyophilized powder; wherein: [0346] the alpha-hydroxy
carboxylic acid or beta-hydroxy carboxylic acid is citric acid;
[0347] A is 0; [0348] R.sup.a is isobutyl; [0349] R.sup.a1 is
hydrogen, C.sub.1-6 aliphatic,
--(CH.sub.2).sub.m--CH.sub.2--R.sup.B, or
--(CH.sub.2).sub.m--CH(R.sup.5a)--OR.sup.5b; [0350] P is
R.sup.c--C(O)--; [0351] R.sup.c is --R.sup.D; [0352] m is 0 or 1;
[0353] the bulking agent is glycine; and [0354] the buffer is
sodium citrate and citric acid.
[0355] In some embodiments, the pharmaceutical composition
comprises the compound of formula (I), a bulking agent, and a
buffer in a lyophilized powder; wherein: [0356] the compound of
formula (I) is represented by compounds (I-1), (I-15) or (I-18);
[0357] the bulking agent is glycine; and [0358] the buffer is
sodium citrate and citric acid.
[0359] In some embodiments, the pharmaceutical composition
comprises the compound (I-1) in a lyophilized powder. In some other
embodiments, the pharmaceutical composition comprises the compound
(I-1), glycine, sodium citrate, and citric acid in a lyophilized
powder. In yet some other embodiments, the pharmaceutical
composition comprises the compound (I-15) in a lyophilized powder.
In still yet some other embodiments, the pharmaceutical composition
comprises the compound (I-15), glycine, sodium citrate, and citric
acid in a lyophilized powder.
[0360] In some embodiments, the invention provides a unit dose
pharmaceutical composition comprising the compound of formula
(I-1), a bulking agent, and a buffer in a lyophilized powder. In
some embodiments, the unit dose pharmaceutical composition
comprises the compound of formula (I-1), glycine, sodium citrate,
and citric acid in a lyophilized powder.
[0361] In some embodiments, the compound of formula (I-1) is
present in the unit dose pharmaceutical composition in an amount
equivalent on a molar weight basis of about 1 mg to about 10 mg of
the compound of formula (VIII-1). In some embodiments, the compound
of formula (I-1) is present in the unit dose pharmaceutical
composition in an amount equivalent on a molar weight basis of
about 1 mg to about 5 mg of the compound of formula (VIII-1). In
some embodiments, the compound of formula (I-1) is present in the
unit dose pharmaceutical composition in an amount equivalent on a
molar weight basis of 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 4.5mg, or about 5.0
mg of the compound of formula (VIII-1). In some embodiments, the
compound of formula (I-1) is present in the unit dose
pharmaceutical composition in an amount equivalent on a molar
weight basis of about 3.5 mg of the compound of formula
(VIII-1).
[0362] In some embodiments, the amount of glycine present in the
unit dose pharmaceutical composition is about 0.01g to about 0.50
g. In some embodiments, the amount of glycine present in the unit
dose pharmaceutical composition is about 0.03 g to about 0.250 g.
In some embodiments, the amount of glycine present in the unit dose
pharmaceutical composition is about 0.06 g to about 0.125 g.
[0363] In some embodiments, the sodium citrate and citric acid is
present in the unit dose pharmaceutical composition in an amount
equivalent to about 0.005 g to about 0.250 g of citrate ion. In
some embodiments, the sodium citrate and citric acid is present in
the unit dose pharmaceutical composition in an amount equivalent to
about 0.025 g to about 0.125 g of citrate ion.
[0364] In some embodiments, the invention provides a unit dose
pharmaceutical composition comprising the compound of formula
(I-15), a bulking agent, and a buffer in a lyophilized powder. In
some embodiments, the unit dose pharmaceutical composition
comprises the compound of formula (I-15), glycine, sodium citrate,
and citric acid in a lyophilized powder.
[0365] In some embodiments, the compound of formula (I-15) is
present in the unit dose pharmaceutical composition in an amount
equivalent on a molar weight basis of about 1 mg to about 10 mg of
the compound of formula (VIII-15). In some embodiments, the
compound of formula (I-15) is present in the unit dose
pharmaceutical composition in an amount equivalent on a molar
weight basis of about 1 mg to about 5 mg of the compound of formula
(VIII-15). In some embodiments, the compound of formula (I-15) is
present in the unit dose pharmaceutical composition in an amount
equivalent on a molar weight basis of 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
4.5mg, or about 5.0 mg of the compound of formula (VIII-15). In
some embodiments, the compound of formula (I-15) is present in the
unit dose pharmaceutical composition in an amount equivalent on a
molar weight basis of about 3.5 mg of the compound of formula
(VIII-15).
[0366] In some embodiments, the amount of glycine present in the
unit dose pharmaceutical composition is about 0.01g to about 0.50
g. In some embodiments, the amount of glycine present in the unit
dose pharmaceutical composition is about 0.03 g to about 0.250 g.
In some embodiments, the amount of glycine present in the unit dose
pharmaceutical composition is about 0.06 g to about 0.125 g.
[0367] In some embodiments, the sodium citrate and citric acid is
present in the unit dose pharmaceutical composition in an amount
equivalent to about 0.005 g to about 0.250 g of citrate ion. In
some embodiments, the sodium citrate and citric acid is present in
the unit dose pharmaceutical composition in an amount equivalent to
about 0.025 g to about 0.125 g of citrate ion.
[0368] In another aspect, the invention provides a method for
preparing the compound of formula (I) as a lyophilized powder; the
method comprising the steps: [0369] (d-1) combining: [0370] i. an
aqueous solvent mixture; [0371] ii. the compound of formula (I);
[0372] iii. a bulking agent; and [0373] iv. a buffer; to form a
mixture; and [0374] (d-2) lyophilizing the mixture.
[0375] In some embodiments, the compound of formula (I) is formed
in situ from the corresponding compound of formula (VIII). Thus,
the invention also provides a method for preparing the compound of
formula (I) as a lyophilized powder; the method comprising the
steps: [0376] (e-1) combining: [0377] i. an aqueous solvent
mixture; [0378] ii. the compound of formula (VIII); [0379] iii. a
bulking agent; and [0380] iv. an alpha-hydroxy carboxylic acid, or
a salt thereof or a beta-hydroxy carboxylic acid, or a salt thereof
or a combination thereof to form a mixture; and [0381] (e-2)
lyophilizing the mixture.
[0382] 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.
[0383] In some embodiments, the aqueous solvent mixture comprises
about 1% v/v to about 40% v/v alcohol. In some other embodiments,
the aqueous solvent mixture comprises about 3% v/v to about 10% v/v
alcohol. In some other embodiments, the aqueous solvent mixture
comprises about 3% v/v to about 6% v/v alcohol. In yet some other
embodiments, the solvent mixture comprises about 3% v/v to about 6%
v/v tert-butyl alcohol. In still yet some other embodiments, the
solvent mixture comprises about 5% v/v tert-butyl alcohol.
[0384] In some embodiments, a method is provided for preparing the
compound (I-1) as a lyophilized powder, the method comprising the
steps: [0385] (f-1) combining: [0386] i. water; [0387] ii. the
compound (I-1); [0388] iii. glycine; [0389] iv. sodium citrate; and
[0390] v. citric acid; to form a mixture; and [0391] (f-2)
lyophilizing the mixture.
[0392] In some embodiments, a method is provided for preparing the
compound (I-15) as a lyophilized powder, the method comprising the
steps: [0393] (g-1) combining: [0394] i. an aqueous solvent mixture
comprising water and tert-butyl alcohol; [0395] ii. the compound
(VIII-15); [0396] iii. glycine; [0397] iv. sodium citrate; and
[0398] v. citric acid; to form a mixture; and [0399] (g-2)
lyophilizing the mixture.
[0400] In some other embodiments, for the method described directly
above, the amount of tert-butyl alcohol present in the aqueous
solvent mixture is about 3% v/v to about 6% v/v.
[0401] The lyophilization or freeze-drying can be performed using
any conventional lyophilizers or freeze-dryers. In some
embodiments, the lyophilization comprises the steps: (i) loading
the liquid mixture as prepared above, and freezing; (ii) annealing;
(iii) second freeze cycle; (iv) drying under vacuum; and (v)
secondary drying. The temperatures and times for each step will
depend on the lyophilizer or freeze-dryer that is employed.
[0402] 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%.
[0403] In another aspect, the invention provides a method for the
preparation of a pharmaceutical composition of the compound of
formula (I) as a liquid pharmaceutical dosage form, said method
comprising the step of reconstituting a lyophilized powder of the
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. Following reconstitution, the
liquid pharmaceutical dosage form can contain concentrations of the
compound of formula (I) as described herein.
[0404] In some embodiments, a method is provided for the
preparation of a pharmaceutical composition of the compound (I-1)
as a liquid pharmaceutical dosage form, said method comprising the
step of reconstituting a lyophilized powder of the compound (I-1)
as described herein with an aqueous solvent suitable for
pharmaceutical administration. In some embodiments, a method is
provided for the preparation of a pharmaceutical composition of the
compound (I-1) as a liquid pharmaceutical dosage form, said method
comprising the step of reconstituting a lyophilized powder of the
compound (I-1) as described herein with water for injection, or
normal saline. In some embodiments, a method is provided for the
preparation of a pharmaceutical composition of the compound (I-1)
as a liquid pharmaceutical dosage form, said method comprising the
step of reconstituting a lyophilized powder of the compound (I-1)
as described herein with water for injection.
[0405] In some embodiments, a method is provided for the
preparation of a pharmaceutical composition of the compound (I-15)
as a liquid pharmaceutical dosage form, said method comprising the
step of reconstituting a lyophilized powder of the compound (I-15)
as described herein with an aqueous solvent suitable for
pharmaceutical administration. In some embodiments, a method is
provided for the preparation of a pharmaceutical composition of the
compound (I-15) as a liquid pharmaceutical dosage form, said method
comprising the step of reconstituting a lyophilized powder of the
compound (I-15) as described herein with water for injection, or
normal saline. In some embodiments, a method is provided for the
preparation of a pharmaceutical composition of the compound (I-15)
as a liquid pharmaceutical dosage form, said method comprising the
step of reconstituting a lyophilized powder of the compound (I-15)
as described herein with water for injection.
[0406] Upon reconstitution in a reconstitution solvent, an
equilibrium is established between the compound of formula (I) and
the corresponding boronic acid of formula (VIII). Typically,
equilibrium is reached quickly within about 10-15 minutes after the
addition of the reconstitution solvent. The relative concentrations
of the boronate ester and boronic acid present at equilibrium is
dependent upon the pH of the solution, temperature, and the ratio
of the alpha-hydroxy or beta-hydroxy acid compound to boronic acid
compound.
[0407] In another aspect, the invention provides a liquid
pharmaceutical composition comprising the compound of formula (I),
and additional excipients described herein. In some embodiments,
the liquid pharmaceutical composition is suitable for parenteral
use. In some other embodiments, the liquid pharmaceutical
composition is suitable for oral use.
[0408] In such embodiments, the liquid pharmaceutical composition
comprises the compound of formula (I), a buffer, and optionally a
tonicity modifier.
[0409] In some embodiments, the ratio of the buffer to the compound
of formula (I) is from about 50:1 to about 10:1. In some other
embodiments, the ratio of the buffer to the compound of formula (I)
is from about 30:1 to about 10:1. In yet some other embodiments,
the ratio of the buffer to the compound of formula (I) is about
20:1.
[0410] In some embodiments, the buffer is present in a
concentration of about 45 mM to about 65 mM. In some other
embodiments, the buffer is present in a concentration of about 50
mM to about 60 mM.
[0411] Suitable buffers include sodium citrate, citric acid, and
mixtures thereof. In some embodiments, the buffer is sodium
citrate, and citric acid.
[0412] Suitable tonicity modifiers 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
modifier is propylene glycol. In some other embodiments, the
tonicity modifier is sodium chloride.
[0413] Upon dissolution in a aqueous solvent mixture, an
equilibrium is established between the compound of formula (I) and
the corresponding boronic acid of formula (VIII). Thus, either a
compound of formula (I) or a compound of formula (VIII) may be used
in the preparation of the liquid pharmaceutical composition.
Typically, equilibrium is reached quickly within about 10-15
minutes after the addition of the aqueous solvent mixture. The
relative concentrations of the boronate ester and boronic acid
present at equilibrium is dependent upon the pH of the solution,
temperature, and the ratio of the alpha-hydroxy or beta-hydroxy
acid compound to boronic acid compound. In some embodiments, the
excess alpha-hydroxy or beta-hydroxy acid may act as a stabilizer,
which pushes the equilibrium towards the boronate ester. In some
embodiments, the tonicity modifier may also act as a
stabilizer.
[0414] In some embodiments, the liquid pharmaceutical composition
optionally further comprises a preservative.
[0415] In some embodiments, the liquid pharmaceutical composition
comprises the compound of formula (I), a buffer, and optionally a
tonicity modifier; wherein: [0416] the alpha-hydroxy carboxylic
acid or beta-hydroxy carboxylic acid is citric acid; [0417] A is 0;
[0418] R.sup.a is isobutyl; [0419] R.sup.a1 is hydrogen, C.sub.1-6
aliphatic, --(CH.sub.2).sub.m--CH.sub.2--R.sup.B, or
--(CH.sub.2).sub.m--CH(R.sup.5a)--OR.sup.b; [0420] P is
R.sup.c--C(O)--; [0421] R.sup.c is --R.sup.D; [0422] m is 0 or 1;
[0423] the buffer is sodium citrate and citric acid; and [0424] the
tonicity modifier, when present, is sodium chloride.
[0425] In some embodiments, the liquid pharmaceutical composition
comprises the compound of formula (I), a buffer, and optionally a
tonicity modifier; wherein: [0426] the compound of formula (I) is
represented by compounds (I-1), (I-15) or (I-18); [0427] the buffer
is sodium citrate and citric acid; and [0428] the tonicity
modifier, when present, is sodium chloride.
[0429] In some embodiments, wherein the alpha-hydroxy carboxylic
acid or beta-hydroxy carboxylic acid is citric acid, 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, wherein the
alpha-hydroxy carboxylic acid or beta-hydroxy carboxylic acid is
citric acid, 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), wherein the
compound of formula (I) is the compound (I-1), water, citric acid,
sodium citrate and sodium chloride.
[0430] In such embodiments, wherein the alpha-hydroxy carboxylic
acid or beta-hydroxy carboxylic acid is citric acid, the liquid
pharmaceutical dosage form of the compound of formula (I) has a pH
of between about pH 3 and about pH 7. In certain such embodiments,
the pH is between about pH 4.9 and about pH 6.7. In other certain
such embodiments, the pH is between about pH 5.5 and about pH
6.5.
[0431] In some embodiments, wherein the alpha-hydroxy carboxylic
acid or beta-hydroxy carboxylic acid is citric acid, the liquid
pharmaceutical composition of the compound of formula (I) is
prepared in situ from a stock vehicle solution and the compound of
formula (VIII). In some embodiments, the stock vehicle solution
comprises water, citric acid, sodium citrate and propylene glycol.
In such embodiments, the resulting solution can be further diluted
with stock vehicle solution or with a sodium chloride solution to
generate liquid pharmaceutical compositions of the compound of
formula (I) of desired concentrations.
[0432] In another aspect the invention provides a unit dose liquid
pharmaceutical composition, comprising the compound of formula (I),
a buffer, and optionally a tonicity modifier. In some embodiments,
the unit dose liquid pharmaceutical composition comprises the
compound of formula (I), a buffer, and optionally a tonicity
modifier, wherein the compound of formula (I) is the 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, wherein the compound of
formula (I) is compound (I-1), the compound (I-1) in the unit dose
liquid pharmaceutical composition is present at a concentration of
about 0.5 mg/ml to about 3 mg/ml of the compound of formula
(VIII-1). In yet some other embodiments, wherein the compound of
formula (I) is compound (I-1), the compound (I-1) in the unit dose
liquid pharmaceutical composition is present at a concentration of
about 1 mg/ml of the compound of formula (VIII-1). In still yet
some other embodiments, wherein the compound of formula (I) is the
compound (I-15), the compound (I-15) in the unit dose liquid
pharmaceutical composition is present at a concentration of about 1
mg/ml of the compound of formula (VIII-15).
[0433] In some embodiments in the unit dose liquid pharmaceutical
composition, 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.
In some embodiments in the unit dose liquid pharmaceutical
composition, the sodium citrate and citric acid are present in an
amount equivalent to about 0.025 g to about 0.125 g of citrate
ion.
[0434] In some embodiments in the unit dose liquid pharmaceutical
composition, the sodium chloride is present in an amount of about
0.0045 g to about 0.09 g. In some embodiments in the unit dose
liquid pharmaceutical composition, the sodium chloride is present
in an amount of about 0.01 g to about 0.04 g.
[0435] In some embodiments in the unit dose liquid pharmaceutical
composition, the pharmaceutical composition is stored frozen until
use.
[0436] In another aspect the invention provides a method for
preparing the compound of formula (I), as a unit dose liquid
pharmaceutical composition; the method comprising the steps: [0437]
(h-1) dissolving the buffer in an aqueous solvent; [0438] (h-2)
dissolving the compound of formula (I), or a crystalline form
thereof, in the mixture obtained in step (h-1); [0439] (h-3)
dissolving the toncity modifier in the mixture obtained in step
(h-2); [0440] (h-4) adding further aqueous solvent to the required
batch volume; and [0441] (h-5) filling vials with an amount of the
mixture obtained in step (h-4).
[0442] In some embodiments, the vials are capped after step (h-5).
In some other embodiments, nitrogen is bubbled through the mixture
prior to step (h-5). In yet some other embodiments, after step
(h-5), the liquid in the vials can be overlayed with nitrogen prior
to capping.
[0443] 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, the
alpha-hydroxy acid or beta-hydroxy acid is added in step (h-2). In
some other embodiments, the alpha-hydroxy acid or beta-hydroxy acid
is present in step (h-1) as the buffer.
[0444] The pharmaceutical compositions of the invention preferably
are formulated for administration to a patient having, or at risk
of developing or experiencing a recurrence of, a
proteasome-mediated disorder. The term "patient", as used herein,
means an animal, preferably a mammal, more preferably a human.
Preferred pharmaceutical compositions of the invention are those
formulated for oral, intravenous, or subcutaneous administration.
However, any of the above dosage forms containing a therapeutically
effective amount of a compound of the invention are well within the
bounds of routine experimentation and therefore, well within the
scope of the instant invention. In some embodiments, the
pharmaceutical composition of the 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 being treated.
[0445] By "therapeutically effective amount" is meant an amount
sufficient to cause a detectable decrease in proteasome activity or
the severity of a proteasome-mediated disorder. The amount of
proteasome inhibitor needed will depend on the effectiveness of the
inhibitor for the given cell type and the length of time required
to treat the disorder. It should also be understood that a specific
dosage and treatment regimen for any particular patient will depend
upon a variety of factors, including the activity of the specific
compound employed, the age, body weight, general health, sex, and
diet of the patient, time of administration, rate of excretion,
drug combinations, the judgment of the treating physician, and the
severity of the particular disease being treated. The amount of
additional therapeutic agent present in a composition of this
invention typically will be no more than the amount that would
normally be administered in a composition comprising that
therapeutic agent as the only active agent. Preferably, the amount
of additional therapeutic agent will range from about 50% to about
100% of the amount normally present in a composition comprising
that agent as the only therapeutically active agent.
[0446] In another aspect, the invention provides a method for
treating a patient having, or at risk of developing or experiencing
a recurrence of, a proteasome-mediated disorder. As used herein,
the term "proteasome-mediated disorder" includes any disorder,
disease or condition which is caused or characterized by an
increase in proteasome expression or activity, or which requires
proteasome activity. The term "proteasome-mediated disorder" also
includes any disorder, disease or condition in which inhibition of
proteasome activity is beneficial.
[0447] For example, compounds and pharmaceutical compositions of
the invention are useful in treatment of disorders mediated via
proteins (e.g., NF.kappa.B, p27.sup.Kip, p21.sup.WAF/CIP1, p53)
.sub.whi.sub.ch .sub.are regul.sub.ated b.sub.y proteasome
activity. Relevant disorders include inflammatory disorders (e.g.,
rheumatoid arthritis, inflammatory bowel disease, asthma, chronic
obstructive pulmonary disease (COPD), osteoarthritis, dermatosis
(e.g., atopic dermatitis, psoriasis)), vascular proliferative
disorders (e.g., atherosclerosis, restenosis), proliferative ocular
disorders (e.g., diabetic retinopathy), benign proliferative
disorders (e.g., hemangiomas), autoimmune diseases (e.g., multiple
sclerosis, tissue and organ rejection), as well as inflammation
associated with infection (e.g., immune responses),
neurodegenerative disorders (e.g., Alzheimer's disease, Parkinson's
disease, motor neurone disease, neuropathic pain, triplet repeat
disorders, astrocytoma, and neurodegeneration as result of
alcoholic liver disease), ischemic injury (e.g., stroke), and
cachexia (e.g., accelerated muscle protein breakdown that
accompanies various physiological and pathological states, (e.g.,
nerve injury, fasting, fever, acidosis, HIV infection, cancer
affliction, and certain endocrinopathies)).
[0448] The compounds and pharmaceutical compositions of the
invention are particularly useful for the treatment of cancer. As
used herein, the term "cancer" refers to a cellular disorder
characterized by uncontrolled or disregulated cell proliferation,
decreased cellular differentiation, inappropriate ability to invade
surrounding tissue, and/or ability to establish new growth at
ectopic sites. The term "cancer" includes, but is not limited to,
solid tumors and bloodborne tumors. The term "cancer" encompasses
diseases of skin, tissues, organs, bone, cartilage, blood, and
vessels. The term "cancer" further encompasses primary and
metastatic cancers.
[0449] Non-limiting examples of solid tumors that can be treated
with the disclosed proteasome inhibitors or pharmaceutical
compositions include pancreatic cancer; bladder cancer; colorectal
cancer; breast cancer, including metastatic breast cancer; prostate
cancer, including androgen-dependent and androgen-independent
prostate cancer; renal cancer, including, e.g., metastatic renal
cell carcinoma; hepatocellular cancer; lung cancer, including,
e.g., non-small cell lung cancer (NSCLC), bronchioloalveolar
carcinoma (BAC), and adenocarcinoma of the lung; ovarian cancer,
including, e.g., progressive epithelial or primary peritoneal
cancer; cervical cancer; gastric cancer; esophageal cancer; head
and neck cancer, including, e.g., squamous cell carcinoma of the
head and neck; melanoma; neuroendocrine cancer, including
metastatic neuroendocrine tumors; brain tumors, including, e.g.,
glioma, anaplastic oligodendroglioma, adult glioblastoma
multiforme, and adult anaplastic astrocytoma; bone cancer; and soft
tissue sarcoma.
[0450] Non-limiting examples of hematologic malignancies that can
be treated with the disclosed proteasome inhibitors or
pharmaceutical compositions include acute myeloid leukemia (AML);
chronic myelogenous leukemia (CML), including accelerated CML and
CML blast phase (CML-BP); acute lymphoblastic leukemia (ALL);
chronic lymphocytic leukemia (CLL); Hodgkin's disease (HD);
non-Hodgkin's lymphoma (NHL), including follicular lymphoma and
mantle cell lymphoma; B-cell lymphoma; T-cell lymphoma; multiple
myeloma (MM); Waldenstrom's macroglobulinemia; myelodysplastic
syndromes (MDS), including refractory anemia (RA), refractory
anemia with ringed siderblasts (RARS), (refractory anemia with
excess blasts (RAEB), and RAEB in transformation (RAEB-T); and
myeloproliferative syndromes.
[0451] In some embodiments, the compound or pharmaceutical
compositions of the invention are used to treat a patient having or
at risk of developing or experiencing a recurrence in a cancer
selected from the group consisting of multiple myeloma and mantle
cell lymphoma.
[0452] In some embodiments, the proteasome inhibitor or
pharmaceutical compositions of the invention is administered in
conjunction with another therapeutic agent. The other therapeutic
agent may also inhibit the proteasome, or may operate by a
different mechanism. In some embodiments, the other therapeutic
agent is one that is normally administered to patients with the
disease or condition being treated. The proteasome inhibitor of the
invention may be administered 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 prior to, at the same time as, or following
administration of the proteasome inhibitor of the invention.
[0453] In some embodiments, a proteasome inhibitor of formula (I),
or pharmaceutical composition of the compound of formula (I) is
administered in conjunction with an anticancer agent. As used
herein, the term "anticancer agent" refers to any agent that is
administered to a subject with cancer for purposes of treating the
cancer.
[0454] Non-limiting examples of DNA damaging chemotherapeutic
agents include topoisomerase I inhibitors (e.g., irinotecan,
topotecan, camptothecin and analogs or metabolites thereof, and
doxorubicin); topoisomerase II inhibitors (e.g., etoposide,
teniposide, and daunorubicin); alkylating agents (e.g., melphalan,
chlorambucil, busulfan, thiotepa, ifosfamide, carmustine,
lomustine, semustine, streptozocin, decarbazine, methotrexate,
mitomycin C, and cyclophosphamide); DNA intercalators (e.g.,
cisplatin, oxaliplatin, and carboplatin); DNA intercalators and
free radical generators such as bleomycin; and nucleoside mimetics
(e.g., 5-fluorouracil, capecitibine, gemcitabine, fludarabine,
cytarabine, mercaptopurine, thioguanine, pentostatin, and
hydroxyurea).
[0455] Chemotherapeutic agents that disrupt cell replication
include: paclitaxel, docetaxel, and related analogs; vincristine,
vinblastin, and related analogs; thalidomide, lenalidomide, and
related analogs (e.g., CC-5013 and CC-4047); protein tyrosine
kinase inhibitors (e.g., imatinib mesylate and gefitinib);
proteasome inhibitors (e.g., bortezomib); NF-.kappa.B inhibitors,
including inhibitors of I.kappa.B kinase; antibodies which bind to
proteins overexpressed in cancers and thereby downregulate cell
replication (e.g., trastuzumab, rituximab, cetuximab, and
bevacizumab); and other inhibitors of proteins or enzymes known to
be upregulated, over-expressed or activated in cancers, the
inhibition of which downregulates cell replication.
[0456] In order that this invention be more fully understood, the
following preparative and testing examples are set forth. These
examples illustrate how to make or test specific compounds, and are
not to be construed as limiting the scope of the invention in any
way.
EXAMPLES
Abbreviations
[0457] DCM methylene chloride [0458] DIPEA N,N'-diisopropylethyl
amine [0459] DMF N,N'-dimethylformamide [0460] EDCI N-(3-dime
thylaminopropyl)-N'-ethylcarbodiimide hydrochloride [0461] EtOAc
ethyl acetate [0462] h hours [0463] HPLC high performance liquid
chromatography [0464] MIBK methyl isobutyl ketone [0465] PES
polyethersulfone [0466] TBTU
O-benzotriazol-1-yl-N,N,N',N'-tetramethyluronium tetrafluoroborate
[0467] TFA trifluoroacetic acid [0468] THF tetrahydrofuran [0469]
HOBt 1-hydroxybenztriazole [0470] LCMS liquid chromatography mass
spectrum [0471] min minutes
General Methods
[0472] .sup.1H NMR: The spectra are acquired at ambient temperature
on a JOEL ECX-400 NMR spectrometer operating at 400 MHz for .sup.1H
NMR. The resulting FID's are transferred to a PC and processed
using NUTS NMR processing software from Acorn NMR Inc. Chemical
shifts are referenced to DMSO solvent, 2.50 ppm. A solvent blank is
prepared by adding .about.0.75 mL of DMSO-d6 to the NMR tube. After
a .sup.1H spectrum is acquired on the solvent blank, the sample is
added and completely dissolved.
[0473] Mass Spectrometry: Mass spectrometry studies are run 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, sheath gas flow rate of 35 arb,
capillary temperature of 275.degree. C., capillary voltage of 9 V
and tube lens offset of 35 V. An analyte was dissolved in
acetonitrile to generate a 0.5 mg/ml solution. An Agilent 1100 HPLC
system was used for LC-Mass spectrometry flow analysis. The pump
flow rate was 1.0 ml/minute. 10 .mu.l of each sample solution was
injected from the autosampler into a T-joint. About 2% of the
solution from the T-joint was infused into the mass
spectrometer.
[0474] X-Ray Powder Diffractometry (XRPD): X-ray powder diffraction
patterns are acquired on either: [0475] i) a Bruker AXS D8Advance
diffractometer. The data is collected over an angular range of
2.9.degree. to 29.6.degree. 2.theta. in continuous scan mode using
a step size of 0.05.degree. 2.theta. and a step time of 2 seconds.
The sample is run under ambient conditions and prepared as a flat
plate specimen using powder as received without grinding; or [0476]
ii) a PANalytical X'Pert Pro diffractometer. Each specimen is
analyzed using Cu radiation produced using an Optix long fine-focus
source. An elliptically graded multilayer mirror is used to focus
the Cu K .alpha. X-rays of the source through the specimen and onto
the detector. The specimen is sandwiched between 3-micron thick
films, analyzed in transmission geometry, and rotated to optimize
orientation statistics. A beam-stop is used to minimize the
background generated by air scattering. Helium and the anti-scatter
extension are not used. Soller slits are used for the incident and
diffracted beams to minimize axial divergence. The diffraction
patterns are collected using a scanning position-minimize axial
divergence. The diffraction patterns are collected using a scanning
position-sensitive detector (X'Celerator) located 240 mm from the
specimen. Prior to the analyses, a silicon specimen (NIST standard
reference material 640c) is analyzed to verify the position of the
silicon 111 peak.
[0477] Differential Scanning calorimetry (DSC): Differential
scanning calorimetry (DSC) data are collected on either: [0478] i)
a TA Instruments Q100 differential scanning calorimeter equipped
with a 50 position auto-sampler. The energy and temperature
calibration standard is indium. Samples are heated at a rate of
10.degree. C. per minute between 25.degree. C. and 300.degree. C. A
nitrogen purge flowing at 50 mL per minute is maintained over the
sample during a scan. Between 1 mg and 3 mg of sample is analyzed.
All samples are crimped in a hermetically sealed aluminum pan with
a pinhole to alleviate the pressure accumulated from the solvent
vapor; or [0479] ii) a TA Instruments differential scanning
calorimeter 2920. The sample is placed into 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.degree.
C., and heated under a nitrogen purge at a rate of 10.degree.
C./min. Indium metal was used as the calibration standard.
[0480] Thermal Gravimetric Analysis (TGA): Thermal gravimetric
analysis (TGA) data are collected on a TA Instruments Q500 thermal
gravimetric analyzer, calibrated with Nickel/Alumel and running at
a scan rate of 10.degree. C. per minute. A nitrogen purge flowing
at 60 mL per minute is maintained over the sample during
measurements. Typically 5 mg to 15 mg of sample is loaded onto a
pre-tared platinum crucible.
Example 1
Synthesis of
4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-m-
ethylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid (I-1)
##STR00017##
[0481] Step 1: 2,5-[(dichlorobenzoyl)amino]acetic acid
[0482] To a mixture of NaOH (12 g, 300 mmol) and glycine (18 g, 239
mmol) in water (120 mL) was added dropwise over 45 min a solution
of 2,5-dichlorobenzoyl chloride (10 g, 48 mmol) in THF (15 mL)
keeping the internal temperature below about 25.degree. C. After 1
h, the mixture was acidified with 2.0 M HCl (125 mL) keeping the
internal temperature below about 5.degree. C. The resulting
precipitate was collected by vacuum filtration. The crude product
was recrystallized from water to give
2,5-[(dichlorobenzoyl)amino]acetic acid as a white, crystalline
solid (6.1 g, 52%). mp 173.3.degree. C. .sup.1H NMR (300 MHz,
DMSO-d.sub.6, .delta.): 12.72 (bs, 1H), 8.89 (t, J=6.0 Hz, 1H),
7.54 (m, 2H), 7.48 (m, 1H), 3.93 (d, J=6.0 Hz). .sup.13C NMR (75
MHz, DMSO-d.sub.6, .delta.): 41.6, 129.3, 129.6, 131.4, 132.2,
138.2, 171.4, 165.9. MS (m/z): [M+H] calculated for
C.sub.9H.sub.8Cl.sub.2NO.sub.3, 248.0; found, 248.0; [M+Na]
calculated for C.sub.9H.sub.7C.sub.2NNaO.sub.3, 270.0; found
270.2.
[0483] 2,5-[(dichlorobenzoyl)amino]acetic acid was also be prepared
via the following procedure: To a mixture of glycine (21.5 g, 286
mmol) in water (437 mL), was added 2.0 M NaOH (130 mL) and the
resulting solution was cooled to 0.degree. C. A solution of
2,5-dichlorobenzoyl chloride (50.0 g, 239 mmol) in THF (75 mL) was
added dropwise at such a rate that the internal temperature was
maintained at 0.+-.1.degree. C. During the addition, the pH was
controlled at 11.0.+-.0.2 using a pH controller titrated with 2.0 M
NaOH. After complete addition, the mixture was stirred at
0.+-.1.degree. C. for an additional 2 h. 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 at 45.degree. C. in a vacuum oven to
afford 2,5-[(dichlorobenzoyl)amino]acetic acid as a white solid
(57.6 g, 97.3%).
Step 2:
2,5-dichloro-N-[2-({(1R)-3-methyl-1-[(3aS,4S,6S,7aR)-3a,5,5-trimet-
hylhexahydro-4,6-methano-1,32-benzodioxaborol-2-yl]butyl}amino)-2-oxoethyl-
]benzamide
[0484] 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.degree. C. was added
(1R)-3-methyl-1-[(3aS,4S,6S,7aR)-3a,5,5-trimethylhexahydro-4,6-methano-1,-
3,2-benodioxaborol-2-yl]butan-1-amine.circle-solid.TFA (9.35 g,
24.7 mmol). DIPEA (13 mL, 75 mmol) was then added dropwise over 2 h
keeping the internal temperature below about 5.degree. C. After 40
min, the mixture was diluted with EtOAc (90 mL), washed with 5%
NaCl (150 mL), twice with 10% NaCl (2.times.40 mL), once with 2%
K.sub.2CO.sub.3 (1.times.40 mL), once with 1% H.sub.3PO.sub.4
(1.times.40 mL), and once with 10% NaCl (1.times.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-trim-
ethylhexahydro-4,6-methano-1,3,2-benzodioxaborol-2-yl]butyl}amino)-2-oxoet-
hyl]benzamide as a white solid which was used in the next step
without purification.
Step 3:
N,N',N''-{boroxin-2,4,6-triyltris[[(1R)-3-methylbutane-1,1-diyl]im-
ino(2-oxoethane-2,1-diyl)]}tris(2,5-dichlorobenzamide)
[0485] To a solution of
2,5-dichloro-N-[2-({(1R)-3-methyl-1-[(3aS,4S,6S,7aR)-3a,5,5-trimethylhexa-
hydro-4,6-methano-1,3,2-benzodioxaborol-2-yl]butyl}amino)-2-oxoethyl]benza-
mide (12.2 g, 24.6 mmol) in methanol/hexane (1:1) (250 mL) were
added 1N 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.times.55 mL). The resulting organic
layer was concentrated to about 10 mL and partitioned between 2.0M
NaOH (30 mL) and DCM (25 mL). The DCM layer was washed once with
additional 2.0M NaOH (5 mL). The basic aqueous layers were then
combined, washed twice with DCM (2.times.25 mL) and acidified with
1M 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.times.10 mL). The combined DCM
extracts were dried over MgSO.sub.4 (25 g) and evaporated to a
thick oil. The product was precipitated with heptane (50 mL) and
collected by filtration to yield
N,N',N''-{boroxin-2,4,6-triyltris[[(1R)-3-methylbutane-1,1-diyl]-
imino(2-oxoethane-2,1-diyl)11tris(2,5-dichlorobenzamide) as a white
solid (6.6 g, 74%). .sup.1H NMR (300 MHz, DMSO-d.sub.6, .delta.):
8.93 (t, J=6.0 Hz, 1H), 8.68 (bs, 1H), 7.63 (m, 1H), 7.52 (m, 2H),
4.00 (d, J=6.0 Hz, 2H), 2.62 (m, 1H), 1.59 (m, 1H), 1.33 (m, 1H),
1.24 (m, 1H), 0.81 (d, J=5.9 Hz, 6H). .sup.13C NMR (125 MHz,
DMSO-d.sub.6, .delta.): 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 CH.sub.3CN: [M+H]
calculated for C.sub.42H.sub.52B.sub.3Cl.sub.6N.sub.6O.sub.9,
1027.2; found, 1027.3; [M+Na] calculated for
C.sub.42H.sub.51B.sub.3Cl.sub.6N.sub.6NaO.sub.9, 1049.2; found
1049.5.
Step 4:
4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetami-
do)-3-methylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid
(I-1)
[0486] Form 1: To a solution of citric acid (2.75 g, 14.3 mmol) in
EtOAc (85 mL) with an internal temperature of about 74.degree. C.
was added
N,N',N''-{boroxin-2,4,6-triyltris[[(1R)-3-methylbutane-1,1-diyl]imino(2-o-
xoethane-2,1-diyl)1}tris(2,5-dichlorobenzamide) (5.00 g, 4.87 mmol)
as a solid. The solution was cooled uncontrolled until the internal
temperature was about 25.degree. C. and the mixture was stirred
overnight. The resulting precipitate was collected by filtration to
yield
2,2'-{2-[(1R)-1-({[(2,5-dichlorobenzoyl)amino]acetyl}amino)-3-methylbutyl-
]-5-oxo-1,3,2-dioxaborolane-4,4-diyl}diacetic acid Form 1 as a
crystalline solid (6.65 g, 88%). .sup.1H NMR (500 MHz,
DMSO-d.sub.6, .delta.110.degree. C.): 10.08 (s, 1H), 8.69 (s, 1H),
7.61 (s, 1H), 7.52 (d, J=1.3 Hz, 2H), 4.26 (d, J=5.5 Hz, 2H), 2.70
(q, J=14.5 Hz, 4H), 2.70 (bs, 1H), 1.72 (sept, J-6.5 Hz, 1H), 1.42
(ddd, J=5.2 Hz, J=8.6 Hz, J=13.9 Hz, 1H), 1.28 (ddd, J=5.3, J=9.4
Hz, J=14.3 Hz, 1H), 0.91 (dd, J=3.3 Hz, J=6.6 Hz, 6H). MS (m/z) in
CH.sub.3CN: [M+Na] calculated for
C.sub.20H.sub.23BCl.sub.2N.sub.2NaO.sub.9, 539.1; found, 539.1.
[0487] The XRPD data for I-1 Form 1 is shown in FIG. 1 and in Table
1.
TABLE-US-00001 TABLE 1 XRPD Data I-1 Form 1 Angle 2.theta. .degree.
Intensity % 6.441 100 8.304 29.5 10.35 19 11.619 5.1 12.695 13.6
15.077 28.2 16.352 28.7 17.504 16.3 18.231 6 19.086 21.4 20.405
11.7 21.231 7.6 21.916 7.6 25.371 15.2 27.588 6.2
[0488] The Differential Scanning calorimetry (DSC) data for I-1
Form 1 is shown in FIG. 2. The profile is characterized by an
endothermic transition with an onset temperature of 191.8.degree.
C. with a melt of 198.8.degree. C. A second endothermic transition
corresponding to decomposition has an onset temperature of
225.degree. C. These temperatures have an error of.+-.5.degree.
C.
[0489] The Thermal Gravimetric Analysis (TGA) data for I-1 Form 1
is shown in FIG. 2. The profile graphs the percent loss of weight
of the sample as a function of temperature, the temperature rate
change being about 10.degree. C/min. The weight loss represents a
loss of about 0.72% of the weight of the sample as the temperature
is changed from 50.degree. C. to 200.degree. C. These temperatures
have an error of .+-.5.degree. C.
[0490] Form 2: To a solution of citric acid (10.1 g, 52.6 mmol) in
EtOAc (300 mL) with an internal temperature of about 74.degree. C.
was added a solution of
N,N,N''-{boroxin-2,4,6-triyltris[[(1R)-3-methylbutane-1,1-diyl]imino(2-ox-
oethane-2,1-diyl)1}tris(2,5-dichlorobenzamide) (20.0 g, 19.5 mmol)
in EtOAc (60 mL). The solution was cooled slowly (about
0.33.degree. C./min) until the internal temperature was about
60.degree. C. and the mixture was stirred for 3 h. The resulting
slurry was cooled slowly (rate of about 0.12.degree. C./min) until
the internal temperature was about 25.degree. C. and the mixture
was stirred overnight. The resulting precipitate was collected by
filtration to yield
4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-m-
ethylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid Form 2 as a
crystalline solid (26.7g, 98%). .sup.1HNMR (500 MHz, DMSO-d.sub.6,
.delta. 110.degree. C.): 10.08 (s, 1H), 8.69 (s, 1H), 7.61 (s, 1H),
7.52 (d, J=1.3 Hz, 2H), 4.26 (d, J=5.5 Hz, 2H), 2.70 (q, J=14.5 Hz,
4H), 2.70 (bs, 1H), 1.72 (sept, J-6.5 Hz, 1H), 1.42 (ddd, J=5.2 Hz,
J=8.6 Hz, J=13.9 Hz, 1H), 1.28 (ddd, J=5.3, J=9.4 Hz, J=14.3 Hz,
1H), 0.91 (dd, J=3.3 Hz, J=6.6 Hz, 6H). .sup.13C NMR (100 MHz,
DMSO-d.sub.6, .delta. 100.degree. 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 CH.sub.3CN:
[M+Na] calculated for C.sub.20H.sub.23BCl.sub.2N.sub.2NaO.sub.9,
539.1; found, 539.1.
[0491]
4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamid-
o)-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''-{boroxin-2,4,6-triyltris[[(1R)-3-methylbutane-1,1-diyl]imino(2-o-
xoethane-2,1-diyl)1}tris(2,5-dichlorobenzamide) (40 g, 0.11 mmol)
in THF (80 mL) at 60.degree. C. The solution was then seeded with
Form 2 crystals (400 mg). After stirring for 30 min at 60.degree.
C., EtOAc (400 mL) was added over a period of 9 h. After complete
addition of the EtOAc, the temperature was lowered to 20.degree. C.
over 5 h. The resulting suspension was filtered to collect
4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-m-
ethylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid Form 2 as a
crystalline solid (40 g, 70%).
[0492]
4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamid-
o)-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.
TABLE-US-00002 TABLE 2 Additional conditions for preparation of I-1
Form 2 Initial Seed Isolated Yield Solvent Temperature Temperature
I-1 Form 2 acetonitrile 80.degree. C. No seeding 77% MIBK
80.degree. C. No seeding 80% 2-methyltetrahydrofuran 80.degree. C.
60.degree. C. 72%
[0493] 4-(R,
S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-methyl-
butyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid Form 2 was also
prepared by dissolving in acetone followed by addition of EtOAc as
an antisolvent.
[0494] The XRPD data for I-1 Form 2 is shown in FIG. 3 and in Table
3.
TABLE-US-00003 TABLE 3 XRPD Data I-1 Form 2 Angle 2-.theta.
.degree. Intensity % 5.817 100 7.614 93.4 11.575 71.1 11.896 67.1
12.571 24.3 14.43 32.2 16.689 65.8 17.362 17.8 18.232 53.9 19.596
77.6 19.959 63.8 20.376 36.2 20.998 32.2 21.5 40.1 21.764 43.4
22.407 77.6 23.12 33.6 23.901 26.3 24.402 20.4 24.882 19.7 25.764
19.1 26.464 39.5 27.347 21.7 27.65 17.1 27.979 16.4 29.41 20.4
[0495] The Differential Scanning calorimetry (DSC) data for I-1
Form 2 is shown in FIG. 4. The profile is characterized by an
endothermic transition with an onset temperature of 206.5.degree.
C. with a melt of 219.9.degree. C. A second endothermic transition
corresponding to decomposition has an onset temperature of
225.degree. C. These temperatures have an error of .+-.5.degree.
C.
[0496] The Thermal Gravimetric Analysis (TGA) data for I-1 Form 2
is shown in FIG. 4. The profile graphs the percent loss of weight
of the sample as a function of temperature, the temperature rate
change being about 10.degree. C./min. The weight loss represents a
loss of about 1.1% of the weight of the sample as the temperature
is changed from 50.degree. C. to 200.degree. C. These temperatures
have an error of .+-.5.degree. C.
Example 1A
Alternate synthesis of
4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-m-
ethylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid (I-1) Form
2
[0497] A 50-L glass reactor equipped with mechanical stirrer,
dropping funnel, temperature indicator, and heating/cooling control
unit (under nitrogen) 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.degree. C. and a solution resulted.
N,N',N''-{boroxin-2,4,6-triyltris[[(1R)-3-methylbutane-1,1-diyl]imino(2-o-
xoethane-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
under stirring (193 rpm) over a period of 20 min while maintaining
a temperature of 73.degree. C. to 75.degree. C. The stirring was
reduced to 96 rpm and the mixture was cooled as follows: (I) The
mixture was kept at 73.degree. C.-75.degree. C. for 25 min; (2) The
mixture was stepwise cooled to 40.degree. C. at the rate of
approximately 5.degree. C./30 min; (3) The mixture was allowed to
cool uncontrolled overnight to ambient temperature with stirring.
The product was then isolated by filtration, washed on the filter
with 1.2 micron filtered EtOAc (2.times.1.2 kg), and dried under
vacuum at 40-41.degree. C. overnight (22 hours) to give 1.458 kg
(92%) of the title compound. .sup.1H NMR (400 MHz, DMSO-d.sub.6,
.delta.): 12.13 (s, 2H), 10.69 (s, 1H), 9.11 (t, J=5.6 Hz, 1H),
7.66 (t, J=1.2 Hz, 1H), 7.56 (d, J=1.2 Hz, 2H), 4.27 (bs, 2H),
2.9-2.55 (m, 5H), 1.67 (bs, 1H), 1.4-1.15 (bs, 2H), 0.86 (d, J=6.4
Hz, 6H).
[0498] The XRPD data for the compound (I-1) Form 2 is shown in FIG.
7 and in Table 6.
TABLE-US-00004 TABLE 6 Angle 2-.theta. .degree. Intensity % 5.69
100 7.64 66 9.66 4 11.22 23 11.42 51 11.79 37 12.41 15 14.23 15
15.60 6 16.53 32 17.15 4 18.07 31 19.39 55 19.79 41 20.24 21 20.79
15 21.36 20 21.61 22 22.23 63 22.55 14 22.97 20 23.22 7 23.67 10
23.90 7 24.19 10 24.74 7 24.97 3 25.64 8 26.31 24 26.64 10 27.21 7
27.40 7 27.88 5 28.25 4 29.27 11 29.72 10
[0499] The Differential Scanning calorimetry (DSC) data for the
compound (I-1) Form 2 is shown in FIG. 8. The profile is
characterized by two endothermic transitions; the first with a melt
at about 231.3.degree. C., and the second with a melt at about
239.9.degree. C. These temperatures have an error of .+-.5.degree.
C.
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)
[0500] To a solution of glycolic acid (0.041 g, 0.54 mmol) in EtOAc
(2.0 mL) with an internal temperature of about 60.degree. C. was
added a solution of
N,N,N''-{boroxin-2,4,6-triyltris[[(1R)-3-methylbutane-1,1-diyl]imino(2-ox-
oethane-2,1-diyl)]}tris(2,5-dichlorobenzamide) (0.199 g, 0.19 mmol)
in EtOAc (1.0 mL). The solution was cooled uncontrolled until the
internal temperature was about 25.degree. C. and the solvent was
removed by evaporation to yield
2,5-dichloro-N-(2-{[(1R)-3-methyl-1-(4-oxo-1,3,2-dioxaborolan-2-yl)butyl]-
amino}-2-oxoethyl)benzamide as a white solid (0.215 g, 95%). MS
(m/z) in CH.sub.3CN: [M+Et.sub.3N+H] calculated for
C.sub.22H.sub.35BCl.sub.2N.sub.3O.sub.5, 502.2; found, 502.0. MS
(m/z) in CH.sub.3CN: [M-H] calculated for
C.sub.16H.sub.18BCl.sub.2N.sub.2O.sub.5, 399.1; found, 399.0.
Example 3
Synthesis of
{(4S)-2-[(1R)-1-({[(2,5-dichlorobenzoyl)amino]-acetyl}amino)-3-methylbuty-
l]-5-oxo-1,3,2-dioxaborolan-4-yl}acetic acid (I-3)
[0501] To a solution of L-malic acid (0.0958 g, 0.714 mmol) in
EtOAc (2.0 mL) with an internal temperature of about 60.degree. C.
was added a solution of
N,N',N''-{boroxin-2,4,6-triyltris[[(1R)-3-methylbutane-1,1-diyl]imino(2-o-
xoethane-2,1-diyl)]}tris(2,5-dichlorobenzamide) (0.239 g, 0.233
mmol) in EtOAc (1.0 mL). The solution was cooled uncontrolled until
the internal temperature was about 25.degree. C. and the solvent
was removed by evaporation to yield
{(4S)-2-[(1R)-1-({[(2,5-dichlorobenzoyl)amino]-acetyl}amino)-3-methylbuty-
l]-5-oxo-1,3,2-dioxaborolan-4-yl}acetic acid as a white solid
(0.307 g, 96%). MS (m/z) in CH.sub.3CN: [M+Et.sub.3N+H] calculated
for C.sub.24H.sub.37BCl.sub.2N.sub.3O.sub.7, 560.1; found, 560.1.
MS (m/z) in CH.sub.3CN: [M-H] calculated for
C.sub.18H.sub.20BCl.sub.2N.sub.2O.sub.7, 457.1; found, 457.1.
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)
[0502] To a solution of (S)-hexahydromandelic acid (0.0881 g, 0.557
mmol) in EtOAc (2.0 mL) with an internal temperature of about
60.degree. C. was added a solution of
N,N',N''-{boroxin-2,4,6-triyltris[[(1R)-3-methylbutane-1,1-diyl]imino(2-o-
xoethane-2,1-diyl)]}tris(2,5-dichlorobenzamide) (0.200 g, 0.195
mmol) in EtOAc (1.0 mL). The solution was cooled uncontrolled until
the internal temperature was about 25.degree. C. and the solvent
was removed by evaporation to yield
2,5-dichloro-N-[2-({(1R)-1-[(4S)-4-cyclohexyl-5-oxo-1,3,2-dioxaborolan-2--
yl]-3-methylbutyl}amino)-2-oxoethyl]benzamide as a white solid
(0.251 g, 93%). MS (m/z) in CH.sub.3CN: [M+Et.sub.3N+H] calculated
for C.sub.28H.sub.45BCl.sub.2N.sub.3O.sub.5, 584.3; found, 584.1.
MS (m/z) in CH.sub.3CN: [M-H] calculated for
C.sub.22H.sub.28BCl.sub.2N.sub.2O.sub.5, 481.1; found 481.1.
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)
[0503] To a solution of 2-hydroxyisobutyric acid (0.0567 g, 0.545
mmol) in EtOAc (2.0 mL) with an internal temperature of about
60.degree. C. was added a solution of
N,N',N''-{boroxin-2,4,6-triyltris[[(1R)-3-methylbutane-1,1-diyl]imino(2-o-
xoethane-2,1-diyl)]}tris(2,5-dichlorobenzamide) (0.200 g, 0.195
mmol) in EtOAc (1.0 mL). The solution was cooled uncontrolled until
the internal temperature was about 25.degree. C. and the solvent
was removed by evaporation to yield
2,5-dichloro-N-(2-{[(1R)-1-(4,4-dimethyl-5-oxo-1,3,2-dioxaborolan-2-yl)-3-
-methylbutyl]amino}-2-oxoethyl)benzamide as a white solid (0.225 g,
96%). MS (m/z) in CH.sub.3CN: [M+Et.sub.3N+H] calculated for
C.sub.24H.sub.39BCl.sub.2N.sub.3O.sub.5, 530.2; found, 530.0. MS
(m/z) in CH.sub.3CN: [M-H] calculated for
C.sub.18H.sub.22BCl.sub.2N.sub.2O.sub.5, 427.1; found, 427.0.
Example 6
Synthesis of
2,5-dichloro-N-[2-({(1R)-3-methyl-1-[(5R)-4-oxo-5-phenyl-1,3,2-dioxaborol-
an-2-yl]butyl}amino)-2-oxoethyl]benzamide (I-6)
[0504] To a solution of (R)-mandelic acid (0.168 g, 1.10 mmol) in
EtOAc (2.0 mL) with an internal temperature of about 60.degree. C.
was added a solution of
N,N,N''-{boroxin-2,4,6-triyltris[[(1R)-3-methylbutane-1,1-diyl]imino(2-ox-
oethane-2,1-diyl)]}tris(2,5-dichlorobenzamide) (0.382 g, 0.37 mmol)
in EtOAc (1.0 mL). The solution was cooled uncontrolled until the
internal temperature was about 25.degree. C. and the resulting
precipitate was collected by filtration to yield
2,5-dichloro-N-[2-({(1R)-3-methyl-1-[(5R)-4-oxo-5-phenyl-1,3,2-dioxaborol-
an-2-yl]butyl}amino)-2-oxoethyl]benzamide as a white solid (0.343
g, 65%). .sup.1H NMR (300 MHz, DMSO-d.sub.6, .delta.): 10.88 (s,
1H), 9.22 (m, 1H), 7.68-7.27 (m, 8H), 5.15 (s, 1H), 4.33 (d, J=6.0
Hz, 2H), 2.8-2.76 (m, 1H), 1.71-1.62 (m, 1H), 1.50-1.28 (m, 2H),
0.89 (m, 6H). MS (m/z) in CH.sub.3CN: [M+Et.sub.3N+H] calculated
for C.sub.28H.sub.39BC.sub.2N.sub.3O.sub.5B, 578.2; found, 578.1.
MS (m/z) in CH.sub.3CN: [M-H] calculated for
C.sub.22H.sub.22BCl.sub.2N.sub.2O.sub.5, 475.1; found 475.1.
Example 7
Synthesis of
2,5-dichloro-N-[2-({(1R)-3-methyl-1-[(4S)-4-methyl-5-oxo-1,3,2-dioxaborol-
an-2-yl]butyl}amino)-2-oxoethyl]benzamide (I-7)
[0505] To a solution of L-lactic acid (0.675 g, 7.34 mmol) in EtOAc
(3.0 mL) with an internal temperature of about 70.degree. C. was
added a solution of
N,N',N''-{boroxin-2,4,6-triyltris[[(1R)-3-methylbutane-1,1-diyl]imino(2-o-
xoethane-2,1-diyl)]}tris(2,5-dichlorobenzamide) (2.50 g, 2.43 mmol)
in EtOAc (7.5 mL). The solution was cooled uncontrolled until the
internal temperature was about 60.degree. C. After 30 min, heptane
(11.5 mL) was added until the solution became turbid. The
suspension was heated until the internal temperature was at or
about 70.degree. C., at which point a homogenous solution resulted.
The solution was cooled at a rate of 0.17.degree. C./min until the
internal temperature was about 30.degree. C., then cooled
uncontrolled until until the internal temperature was about
0.degree. C. The resulting precipitate was collected by filtration
to yield yield
2,5-dichloro-N-[2-({(1R)-3-methyl-1-[(4S)-4-methyl-5-oxo-1,3,2-dioxaborol-
an-2-yl]butyl}amino)-2-oxoethyl]benzamide as a white, crystalline
solid (2.32 g, 81%). MS (m/z) in CH.sub.3CN: [M+Et.sub.3N+H]
calculated for C.sub.23H.sub.37BCl.sub.2N.sub.3O.sub.5, 515.9;
found, 516.0. MS (m/z) in CH.sub.3CN: [M-H] calculated for
C.sub.17H.sub.20BCl.sub.2N.sub.2O.sub.5, 413.1; found 413.0.
[0506] The XRPD data for I-7 is shown in FIG. 5 and in Table 4.
TABLE-US-00005 TABLE 4 XRP DATA I-7 Angle 2-.theta. .degree.
Intensity % 7.404 46 8.783 63.5 9.402 16.1 11.9 20.6 12.195 100
13.71 7.3 14.594 26.5 15.302 8.3 15.772 31 17.299 26.8 17.859 25.8
18.549 22.7 19.943 55.5 20.214 33.9 20.606 50 21.48 15.6 21.887 23
22.75 30.1 23.028 53.1 23.334 28.9 24.243 18.2 25.2 13.3 25.566
37.7 27.221 10 29.103 9.2 29.383 12.6
Example 8
Synthesis of
2,5-dichloro-N-[2-({(1R)-3-methyl-1-[(4S)-4-methyl-6-oxo-1,3,2-dioxaborin-
an-2-yl]butyl}amino)-2-oxoethyl]benzamide (I-8)
[0507] To a solution of (S)-3-hydroxybutyric acid (0.0598 g, 0.566
mmol) in EtOAc (2.0 mL) with an internal temperature of about
60.degree. C. was added a solution of
N,N',N''-{boroxin-2,4,6-triyltris[[(1R)-3-methylbutane-1,1-diyl]imino(2-o-
xoethane-2,1-diyl)]}tris(2,5-dichlorobenzamide) (0.200 g, 0.195
mmol) in EtOAc (1.0 mL). The solution was cooled uncontrolled until
the internal temperature was about 25.degree. C. and the solvent
was removed by evaporation to yield
2,5-dichloro-N-[2-({(1R)-3-methyl-1-[(4S)-4-methyl-6-oxo-1,3,2-dioxaborin-
an-2-yl]butyl}amino)-2-oxoethyl]benzamide as a white solid (0.225
g, 95%). .sup.1H NMR (300 MHz, DMSO-d.sub.6, .delta.): 10.45 (s,
1H), 9.11 (t, J =6.0 Hz, 1H), 7.65 (m, 1H), 7.55 (m, 2H), 4.21 (d,
J =6.0 Hz, 2H), 3.98-3.90 (m, 1H), 2.51 (m, 1H), 2.33 (dd,
J.sub.1=19.2 Hz, J=2.7 Hz, 1H), 2.24-2.21 (m, 1H), 1.61-1.52 (m,
1H), 1.33-1.19 (m, 2H), 1.07-1.04 (m, 3H), 0.84 (m, 6H). MS (m/z)
in CH.sub.3CN: [M+Et.sub.3N+H] calculated for
C.sub.24H.sub.39BCl.sub.2N.sub.3O.sub.5, 530.2; found, 530.0. MS
(m/z) in CH.sub.3CN: [M-H] calculated for
C.sub.18H.sub.22BCl.sub.2N.sub.2O.sub.5, 427.1; found, 427.1.
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)
[0508] To a solution of .beta.-hydroxyisovaleric acid (0.0841 g,
0.712 mmol) in EtOAc (2.0 mL) with an internal temperature of about
60.degree. C. was added a solution of
N,N',N''-{boroxin-2,4,6-triyltris[[(1R)-3-methylbutane-1,1-diyl]imino(2-o-
xoethane-2,1-diyl)1}tris(2,5-dichlorobenzamide) (0.260 g, 0.253
mmol) in EtOAc (1.0 mL). The solution was cooled uncontrolled until
the internal temperature was about 25.degree. C. and the solvent
was removed by evaporation to yield
2,5-dichloro-N-(2-{[(1R)-1-(4,4-dimethyl-6-oxo-1,3,2-dioxaborinan-2-yl)-3-
-methylbutyllamino}-2-oxoethyl)benzamide as a white solid (0.296 g,
95%). MS (m/z) in CH.sub.3CN: [M+Et.sub.3N+H] calculated for
C.sub.25H.sub.41BCl.sub.2N.sub.3O.sub.5, 544.3; found, 544.0. MS
(m/z) in CH.sub.3CN: [M-H] calculated for
C.sub.19H.sub.24BCl.sub.2N.sub.2O.sub.5, 441.1; found, 441.0.
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)
[0509] To a solution of (S)-2-hydroxy-3,3-dimethylbutyric acid
(0.0712 g, 0.553 mmol) in EtOAc (2.0 mL) with an internal
temperature of about 60.degree. C. was added a solution of
N,N',N''-{boroxin-2,4,6-triyltris[[(1R)-3-methylbutane-1,1-diyl]imino(2-o-
xoethane-2,1-diyl)]}tris(2,5-dichlorobenzamide) (0.200 g, 0.195
mmol) in EtOAc (1.0 mL). The solution was cooled uncontrolled until
the internal temperature was about 25.degree. C. and the solvent
was removed by evaporation to yield
2,5-dichloro-N-[2-({(1R)-1-[(4S)-4-tert-butyl-5-oxo-1,3,2-dioxaborolan-2--
yl]-3-methylbutyl}amino)-2-oxoethyl]-2,5-dichlorobenzamide as a
white solid (0.245 g, 97%). MS (m/z) in CH.sub.3CN: [M+Et.sub.3N+H]
calculated for C.sub.26H.sub.43BCl.sub.2N.sub.3O.sub.5, 558.3;
found, 558.0. MS (m/z) in CH.sub.3CN: [M-H] calculated for
C.sub.20H.sub.26BCl.sub.2N.sub.2O.sub.5, 455.1; found, 455.0.
Example 11
Synthesis of
2,5-dichloro-N-[2-({(1R)-1-[(4S)-4-isopropyl-5-oxo-1,3,2-dioxaborolan-2-y-
l]-3-methylbutyl}amino)-2-oxoethyl]benzamide (I-11)
[0510] To a solution of (S)-2-hydroxy-3-methylbutyric acid (0.0659
g, 0.558 mmol) in EtOAc (2.0 mL) with an internal temperature of
about 60.degree. C. was added a solution of
N,N',N''-{boroxin-2,4,6-triyltris[[(1R)-3-methylbutane-1,1-diyl]imino(2-o-
xoethane-2,1-diyl)]}tris(2,5-dichlorobenzamide) (0.200 g, 0.195
mmol) in EtOAc (1.0 mL). The solution was cooled uncontrolled until
the internal temperature was about 25.degree. C. and the solvent
was removed by evaporation to yield
2,5-dichloro-N-[2-({(1R)-1-[(4S)-4-isopropyl-5-oxo-1,3,2-dioxaborolan-2-y-
l]-3-methylbutyl}amino)-2-oxoethyl]benzamide as a white solid
(0.246 g, 99%). MS (m/z) in CH.sub.3CN: [M+Na] calculated for
C.sub.19H.sub.25BCl.sub.2N.sub.2NaO.sub.5, 465.1; found, 465.1. MS
(m/z) in CH.sub.3CN: [M-H] calculated for
C.sub.19H.sub.24BCl.sub.2N.sub.2O.sub.5, 441.1; found, 441.0.
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)
[0511] To a solution of 2-hydroxyisocaproic acid (0.0752 g, 0.569
mmol) in EtOAc (2.0 mL) with an internal temperature of about
60.degree. C. was added a solution of
N,N',N''-{boroxin-2,4,6-triyltris[[(1R)-3-methylbutane-1,1-diyl]imino(2-o-
xoethane-2,1-diyl)]}tris(2,5-dichlorobenzamide) (0.200 g, 0.195
mmol) in EtOAc (1.0 mL). The solution was cooled uncontrolled until
the internal temperature was about 25.degree. C. and the solvent
was removed by evaporation to yield
2,5-dichloro-N-[2-({(1R)-1-[(4S)-4-isobutyl-5-oxo-1,3,2-dioxaborolan-2-yl-
]-3-methylbutyl}amino)-2-oxoethyl]benzamide as a white solid (0.253
g, 95%). MS (m/z) in CH.sub.3CN: [M+Na] calculated for
C.sub.20H.sub.27BCl.sub.2N.sub.2Na.sub.1O.sub.5, 479.1; found,
479.1. MS (m/z) in CH.sub.3CN: [M-H] calculated for
C.sub.20H.sub.26BCl.sub.2N.sub.2O.sub.5, 455.1; found 455.1.
Example 13
Synthesis of
2,5-dichloro-N-(2-{[(1R)-3-methyl-1-(4-oxo-4H-1,3,2-benzodioxaborinin-2-y-
l)butyl]amino}-2-oxoethyl)benzamide (I-13)
[0512] To a solution of salicylic acid (0.0758 g, 0.549 mmol) in
EtOAc (2.0 mL) with an internal temperature of about 60.degree. C.
was added a solution of
N,N',N''-{boroxin-2,4,6-triyltris[[(1R)-3-methylbutane-1,1-diyl]imino(2-o-
xoethane-2,1-diyl)]}tris(2,5-dichlorobenzamide) (0.200 g, 0.195
mmol) in EtOAc (1.0 mL). The solution was cooled uncontrolled until
the internal temperature was about 25.degree. C. and the resulting
precipitate was collected by filtration to yield
2,5-dichloro-N-(2-{[(1R)-3-methyl-1-(4-oxo-4H-1,3,2-benzodioxaborinin-2-y-
l)butyl]amino}-2-oxoethyl)benzamide as a white solid (0.198 g,
78%). MS (m/z) in CH.sub.3CN: [M+Na] calculated for
C.sub.21H.sub.21BCl.sub.2N.sub.2NaO.sub.5, 485.1; found, 485.1. MS
(m/z) in CH.sub.3CN: [M-H] calculated for
C.sub.21H.sub.20BCl.sub.2N.sub.2O.sub.5, 461.1; found, 461.0.
[0513] The XRPD data for I-13 is shown in FIG. 6 and in Table
5.
TABLE-US-00006 TABLE 5 XRP DATA I-13 Angle 2-.theta. .degree.
Intensity % 6.784 88.1 8.372 100 11.855 66.6 13.18 85.2 14.118 7.7
14.546 19.3 15.614 9.6 16.123 19.3 16.417 14.1 16.738 7.7 17.29
43.7 19.05 17.4 19.28 28.9 19.726 52.1 20.401 60.8 20.591 37.6
21.233 43.7 21.658 16.7 22.029 18.6 22.718 30.9 23.557 41.5 24.236
22.2 24.717 62.1 25.309 26 25.648 13.5 26.186 69.1 26.653 17.4
26.995 36.3 27.956 25.4 28.898 8.4 29.47 8.7
Example 14
Synthesis of
2,5-dichloro-N-(2-{[(1R)-3-methyl-1-(5-oxo-4,4-diphenyl-1,3,2-dioxaborola-
n-2-yl)butyl]amino}-2-oxoethyl)benzamide (I-14)
[0514] To a solution of benzilic acid (0.126 g, 0.552 mmol) in
EtOAc (2.0 mL) with an internal temperature of about 60.degree. C.
was added a solution of
N,N',N''-{boroxin-2,4,6-triyltris[[(1R)-3-methylbutane-1,1-diyl]imino(2-o-
xoethane-2,1-diyl)]}tris(2,5-dichlorobenzamide) (0.200 g, 0.195
mmol) in EtOAc (1.0 mL). The solution was cooled uncontrolled until
the internal temperature was about 25.degree. C. and the solvent
was removed by evaporation to yield
2,5-dichloro-N-(2-{[(1R)-3-methyl-1-(5-oxo-4,4-diphenyl-1,3,2-dioxaborola-
n-2-yl)butyl]amino}-2-oxoethyl)benzamide as a white solid (0.291 g,
95%). MS (m/z) in CH.sub.3CN: [M+Na] calculated for
C.sub.28H.sub.27BCl.sub.2N.sub.2NaO.sub.5, 575.1; found, 575.2. MS
(m/z) in CH.sub.3CN: [M-H] calculated for
C.sub.28H.sub.26BCl.sub.2N.sub.2O.sub.5, 551.1; found, 551.1.
Example 15
Synthesis of
2,2'-{2-[(1R)-3-methyl-1-({(2S)-3-phenyl-2-[(pyrazin-2-ylcarbonyl)amino]p-
ropanoyl}amino)butyl]-5-oxo-1,3,2-dioxaborolane-4,4-diyl}diacetic
acid (I-15)
[0515] To a solution of citric acid (0.257 g, 1.34 mmol) in EtOAc
(7.4 mL) with an internal temperature of about 74.degree. C. was
added
N,N',N''-(boroxin-2,4,6-triyltris{[(1R)-3-methylbutane-1,1-diyl]iminoR2S)-
-1-oxo-3-phenylpropane-1,2-diyl]})tripyrazine-2-carboxamide (0.500
g, 0.455 mmol) as a solid. The resulting solution was cooled
uncontrolled until the internal temperature was about 25.degree. C.
and was evaporated to yield
2,2'-{2-[(1R)-3-methyl-1-({(2S)-3-phenyl-2-[(pyrazin-2-ylcarbony-
l)amino]propanoyl}amino)butyl}-5-oxo-1,3,2-dioxaborolane-4,4-diyl}diacetic
acid as a white solid (0.730 g, 99%). MS (m/z) in CH.sub.3CN:
[M+Et.sub.3N+H] calcd for C.sub.31H.sub.45BN.sub.5O.sub.9, 642.3;
found, 642.2. MS (m/z) in CH.sub.3CN: [M-H] calcd for
C.sub.25H.sub.28BN.sub.4O.sub.9, 539.2; found, 539.2.
Example 16
Synthesis of
N-[(1S)-1-benzyl-2-({(1R)-3-methyl-1-[(5R)-4-oxo-5-phenyl-1,3,2-dioxaboro-
lan-2-yl]butyl}amino)-2-oxoethyl]pyrazine-2-carboxamide (I-16)
[0516] To a solution of (R)-mandelic acid (0.0738 g, 0.485 mmol) in
EtOAc (2.0 mL) with an internal temperature of about 60.degree. C.
was added
N,N',N''-(boroxin-2,4,6-triyltris{[(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 cooled uncontrolled
until the internal temperature was about 25.degree. C. and the
resulting precipitate was collected by filtration to yield
N-[(1S)-1-benzyl-2-((1R)-3-methyl-1-[(5R)-4-oxo-5-phenyl-1,3,2-dioxaborol-
an-2-yl]butyl}amino)-2-oxoethyl]pyrazine-2-carboxamide as a white
solid (0.195 g, 80%). MS (m/z) in CH.sub.3CN: [M+Na] calculated for
C.sub.27H.sub.29BN.sub.4NaO.sub.5, 523.2; found, 523.2. MS (m/z) in
CH.sub.3CN: [M-H] calculated for C.sub.27H.sub.28BN.sub.4O.sub.5,
499.2; found, 499.2.
Example 17
Synthesis of
N-[(1S)-1-benzyl-2-({(1R)-3-methyl-1-[(5R)-4-oxo-5-phenyl-1,3,2-dioxaboro-
lan-2-yl]butyl}amino)-2-oxoethyl]pyrazine-2-carboxamide (I-17)
[0517] To a solution of (S)-3-hydroxybutyric acid (0.0509 g, 0.489
mmol) in EtOAc (2.0 mL) with an internal temperature of about
60.degree. C. was added
N,N',N''-(boroxin-2,4,6-triyltris{[(1R)-3-methylbutane-1,1-diyl]imi-
no R2 S)-1-oxo-3-phenylpropane-1,2-diyl]})tripyrazine-2-carboxamide
(0.179 g, 0.163 mmol) as a solid. The solution was cooled
uncontrolled until the internal temperature was about 25.degree. C.
and the solvent was removed by evaporation to yield
N-[(1S)-1-benzyl-2-({(1R)-3-methyl-1-[(4S)-4-methyl-6-oxo-1,3,2-dioxabori-
nan-2-yl]butyl}amino)-2-oxoethyl]pyrazine-2-carboxamide as a white
solid (0.213 g, 96%). MS (m/z) in CH.sub.3CN: [M+Na] calculated for
C.sub.23H.sub.29BN.sub.4NaO.sub.5, 475.2; found, 475.2. MS (m/z) in
CH.sub.3CN: [M-H] calculated for C.sub.23H.sub.28BN.sub.4O.sub.5,
451.2; found, 451.1.
Example 18
Preparation of formulations of
4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-m-
ethylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid (I-1) for
parenteral or oral administration
[0518] Formulation A: A vessel was charged with 90 mL water and
citric acid monohydrate (0.08 g) and sodium citrate dihydrate (1.5
g) were added and stirred until dissolved. To this solution,
4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-m-
ethylbutyl)-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. To this solution, sodium chloride (0.45 g) was added,
and the pH was adjusted to pH 5.45 using 2N HCl. The final volume
of the resulting solution was adjusted to 100 mL with water and
filtered through a 0.2 .mu.m PES membrane to yield Formulation A
which was stored at -20.degree. C.
[0519] Formulation B was prepared as for Formulation A, except that
the pH was adjusted to pH 6.2 using 2N NaOH.
[0520] Formulation C: A vessel was charged with 90 mL 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. To this solution,
4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acet-
amido)-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. The pH was adjusted to 6.2 using 2N NaOH,
and the final volume of the resulting solution was adjusted to 100
mL with water and filtered through a 0.2 .mu.m PES membrane to
yield Formulation C which was stored at -20.degree. C.
Example 19
In situ preparation of formulations of
4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-m-
ethylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid (I-1) for
parenteral or oral administration
[0521] Stock Formulation Vehicle: A vessel was charged with
approximately 160 mL 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 final pH was pH 5.14. The final weight of the
resulting solution was adjusted to 200 g (assuming density of 1
g/mL) with water and filtered through a 0.2 .mu.m PES membrane
filter unit and stored at a temperature between about 2.degree. C.
and about 8.degree. C.
[0522] Formulation Stock (1 mg/mL): To a vessel, 0.105 grams
(approximately 95.4% purity) of N,N',N''-{boroxin-2,4,6-triyltris
[[(1R)-3-methylbutane-1,1-diyl]imino(2-oxoethane-2,1-diyl)]}tris(2,5-dich-
lorobenzamide) was added. To this was added approximately 90 g of
the Stock Formulation Vehicle, and the resulting mixture was
stirred for 48 hours protected from light. The final pH was pH
5.12. The final weight of the resulting solution was adjusted to
100 g (assuming density of 1 g/mL) with Stock Formulation Vehicle
and filtered through a 0.2 .mu.m PES membrane filter unit and
stored protected from light at a temperature between about
2.degree. C. and about 8.degree. C.
[0523] Formulation D: The Formulation Stock was diluted to
concentrations of 0.05 mg/mL and 0.1 mg/mL with Stock Formulation
Vehicle prior to use.
[0524] Formulation E: The Formulation Stock was diluted to
concentrations of 0.05 mg/mL and 0.1 mg/mL with a 0.9% sodium
chloride solution prior to use.
Example 20
20S Proteasome Assay
[0525] To 1 .mu.L of test compound dissolved in DMSO in a 384-well
black microtiter plate is added 25 .mu.L of assay buffer at
37.degree. C. containing human PA28 activator (Boston Biochem, 12
nM final) with Ac-WLA-AMC (.beta.5 selective substrate) (15 .mu.M
final), followed by 25 .mu.L of assay buffer at 37.degree. C.
containing human 20S proteasome (Boston Biochem, 0.25 nM final).
Assay buffer is composed of 20 mM HEPES, 0.5 mM EDTA and 0.01% BSA,
pH7.4. The reaction is followed on a BMG Galaxy plate reader
(37.degree. C., excitation 380 nm, emission 460 nm, gain 20).
Percent inhibition is calculated relative to 0% inhibition (DMSO)
and 100% inhibition (10 .mu.M bortezomib) controls.
Example 21
Antiproliferation Assay
[0526] HCT-116 (1000) or other tumor cells in 100 .mu.L of
appropriate cell culture medium (McCoy's 5A for HCT-116,
Invitrogen) supplemented with 10% fetal bovine serum (Invitrogen)
are seeded in wells of a 96-well cell culture plate and incubated
overnight at 37.degree. C. Test compounds are added to the wells
and the plates are incubated for 96 hours at 37.degree. C. MTT or
WST reagent (10 .mu.L, Roche) are added to each well and incubated
for 4 hours at 37.degree. C. as described by the manufacturer. For
MTT the metabolized dye is solubilized overnight according to
manufacturer's instructions (Roche). The optical density for each
well is read at 595 nm (primary) and 690 nm (reference) for the MTT
and 450 nm for the WST using a spectrophotometer (Molecular
Devices). For the MTT the reference optical density values are
subtracted from the values of the primary wavelength. Percent
inhibition is calculated using the values from a DMSO control set
to 100%.
Example 22
In vivo Tumor Efficacy Model
[0527] Freshly dissociated HCT-116 (2-5.times.10.sup.6) or other
tumor cells in 100 .mu.L of RPMI-1640 media (Sigma-Aldrich) are
aseptically injected into the subcutaneous space in the right
dorsal flank of female CD-1 nude mice (age 5-8 weeks, Charles
River) using a 1 mL 26 3/8-ga needle (Becton Dickinson Ref
#309625). Alternatively, some xenograft models require the serial
passaging of tumor fragments. In these cases, small fragments of
tumor tissue (approximately 1 mm.sup.3) are implanted
subcutaneously in the right dorsal flank of anesthetized (3-5%
isoflourane/oxygen mixture) C.B-17/SCID mice (age 5-8 weeks,
Charles River) via a 13-ga trocar (Popper & Sons 7927).
Beginning at day 7 after inoculation tumors are measured twice
weekly using a vernier caliper. Tumor volumes are calculated using
standard procedures (0.5.times.(length.times.width.sup.2)). When
the tumors reach a volume of approximately 200 mm.sup.3 mice are
randomized into treatment groups and begin receiving drug
treatment. Dosing and schedules are determined for each experiment
based on previous results obtained from
pharmacokinetic/pharmacodynamic and maximum tolerated dose studies.
The control group will receive vehicle without any drug. Typically,
test compound (100-200 .mu.L) is administered via intravenous
(27-ga needle), oral (20-ga gavage needle) or subcutaneous (27-ga
needle) routes at various doses and schedules. Tumor size and body
weight are measured twice a week and the study is terminated when
the control tumors reach approximately 2000 mm.sup.3.
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)pyrazin-
e-2-carboxamide (I-19)
[0528] A mixture of N,N',N''-(boroxin-2,4,6-triyltris
[(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 salicyclic acid (269.6 mg, 0.68 mmol)
were mixed in EtOAc (10 mL). The mixture was heated to form a
solution. The solution was cooled uncontrolled until the internal
temperature was about 25.degree. C. Heptane (16 mL) was added.
White solid precipitated out and the resultant slurry was agitated
at ambient temperature for 3 h. The slurry was filtered to collect
solid
N--((S)-1-((R)-3-methyl-1-(4-oxo-4H-benzo[d][1,3,2]dioxaborinin-2-yl)buty-
lamino)-1-oxo-3-phenylpropan-2-yl)pyrazine-2-carboxamide (0.249 g,
75%). MS (m/z) in CH.sub.3CN: [M+H] calculated for
C.sub.26H.sub.28BN.sub.4O.sub.5, 487.2153; found, 487.3.
Example 24
Synthesis of
2-((S)-2-((R)-3-methyl-1-((S)-3-phenyl-2-(pyrazine-2-carboxamido)propanam-
ido)butyl)-5-oxo-1,3,2-dioxaborolan-4-yl)acetic acid (I-20)
[0529] A mixture of
N,N',N''-(boroxin-2,4,6-triyltris{[(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) were mixed in
THF (5 mL). The mixture was heated to form a solution. The solution
was cooled uncontrolled until the internal temperature was about
25.degree. C. White solid precipitated out and the resultant slurry
was agitated at ambient temperature for 1 h. The slurry was
filtered to collect solid
2-((S)-2-((R)-3-methyl-1-((S)-3-phenyl-2-(pyrazine-2-carboxamido)propanam-
ido)butyl)-5-oxo-1,3,2-dioxaborolan-4-yl)acetic acid (0.625 g,
95%). MS (m/z) in CH.sub.3CN: [M+H] calculated for
C.sub.23H.sub.28BN.sub.4O.sub.7, 483.2051; found, 483.2.
Example 25
Synthesis of
2-((R)-2-((R)-3-methyl-1-((S)-3-phenyl-2-(pyrazine-2-carboxamido)propanam-
ido)butyl)-5-oxo-1,3,2-dioxaborolan-4-yl)acetic acid (I-21)
[0530] A mixture of N,N',N''-(boroxin-2,4,6-triyltris
[(1R)-3-methylbutane-1,1-diyl]imino[(2S)-1-oxo-3-phenylpropane-1,2-diyl]}-
)tripyrazine-2-carboxamide (0.305 g, 0.278 mmol) and D-malic acid
(130.3 mg, 0.33 mmol) were mixed in acetone (3 mL). The mixture was
heated to form a solution. The solution was cooled uncontrolled
until the internal temperature was about 25.degree. C. White solid
precipitated out and the resultant slurry was agitated at ambient
temperature for 3 h. The slurry was filtered to collect solid
2-((R)-2-((R)-3-methyl-1-((S)-3-phenyl-2-(pyrazine-2-carboxamido)propanam-
ido)butyl)-5-oxo-1,3,2-dioxaborolan-4-yl)acetic acid (0.410 g,
100%). [M+H] calculated for C.sub.23H.sub.28BN.sub.4O.sub.7,
483.2051; found, 483.2.
Example 26
Synthesis of
(R)-2-hydroxy-2-((R)-2-((R)-3-methyl-1-((S)-3-phenyl-2-(pyrazine-2-carbox-
amido)propanamido)butyl)-5-oxo-1,3,2-dioxaborolan-4-yl)acetic acid
(I-22)
[0531] A mixture of
N,N',N''-(boroxin-2,4,6-triyltris{[(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) were mixed
in acetone (3 mL). The mixture was heated to form a solution. The
solution was cooled uncontrolled until the internal temperature was
about 25.degree. C. Heptane (2.5 mL) was added.
[0532] White solid precipitated out and the resultant slurry was
agitated at ambient temperature for 1.5 h. The slurry was filtered
to collect 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) which also contained a dimeric species. MS (m/z) in
CH.sub.3CN: [M+H] calculated for C.sub.23H.sub.28BN.sub.4O.sub.8,
499.2000; found, 499.2.
Example 27
Synthesis of
(S)-2-hydroxy-2-((S)-2-((R)-3-methyl-1-((S)-3-phenyl-2-(pyrazine-2-carbox-
amido)propanamido)butyl)-5-oxo-1,3,2-dioxaborolan-4-yl)acetic acid
(I-23)
[0533] A mixture of
N,N',N''-(boroxin-2,4,6-triyltris{[(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) were mixed
in acetone (4 mL). The mixture was heated to form a solution. The
solution was cooled uncontrolled until the internal temperature was
about 25.degree. C. Heptane (8 mL) was added. The mixture was
evaporated to yield
(S)-2-hydroxy-2-((S)-2-((R)-3-methyl-1-((S)-3-phenyl-2-(pyrazine-2--
carboxamido)propanamido)butyl)-5-oxo-1,3,2-dioxaborolan-4-yl)acetic
acid (0.447 g) which also contained a dimeric species. MS (m/z) in
CH.sub.3CN: [M+H] calculated for C.sub.23H.sub.28BN.sub.4O.sub.8,
499.2000; found, 499.2.
Example 28
Pharmaceutical Composition 1
[0534] The composition of the capsule is shown in Table 7
below.
TABLE-US-00007 TABLE 7 Capsule composition mg/ Component Function
Capsule Compound of formula (I-1) Fonn 2 0.29 Microcrystalline
cellulose (low moisture) Filler 89.71 Total capsule content weight,
mg 90.00 Size 4 white opaque gelatin capsules
Example 29
Pharmaceutical Composition 2
[0535] The composition of the capsule is shown in Table 8
below.
TABLE-US-00008 TABLE 8 Capsule composition mg/ Component Function
Capsule Compound of formula (I-1) Fonn 2 0.29 Silicified
microcrystalline cellulose Filler 109.71 Total capsule content
weight, mg 110.00 Size 4 white opaque gelatin capsules
Example 30
Pharmaceutical Composition 3
[0536] The composition of the capsule is shown in Table 9
below.
TABLE-US-00009 TABLE 9 Capsule composition mg/ Component Function
Capsule Compound of formula (I-1) Form 2 0.29 Microcrystalline
cellulose (low moisture) Filler 88.81 Magnesium stearate Lubricant
0.90 Total capsule content weight, mg 90.00 Size 4 white opaque
gelatin capsule
Example 31
Pharmaceutical Composition 4
[0537] The composition of the capsule is shown in Table 10
below.
TABLE-US-00010 TABLE 10 Capsule composition mg/ Component Function
Capsule Compound of formula (I-1) Form 2 0.29 Microcrystalline
cellulose Filler 78.91 Magnesium stearate Lubricant 0.80 Total
capsule content weight, mg 80.00 Size 4 white opaque gelatin
capsule
Example 32
Pharmaceutical Composition 5
[0538] The composition of the capsule is shown in Table 11
below.
TABLE-US-00011 TABLE 11 Capsule composition mg/ Component Function
Capsule Compound of formula (I-1) Form 2 0.29 Microcrystalline
cellulose (low moisture) Filler 84.71 Total capsule contents
weight, mg 85.00 Size 4 white opaque gelatin capsule
Example 33
Pharmaceutical Composition 6
[0539] The composition of the capsule is shown in Table 12
below.
TABLE-US-00012 TABLE 12 Capsule composition mg/ Component Function
Capsule Compound of formula (I-1) Form 2 0.72 Microcrystalline
cellulose (low moisture) Filler 119.28 Total capsule content
weight, mg 120.00 Size 3 dark green gelatin capsule
Example 34
Pharmaceutical Composition 7
[0540] The composition of the capsule is shown in Table 13
below.
TABLE-US-00013 TABLE 13 Capsule composition mg/ Component Function
Capsule Compound of formula (I-1) Form 2 2.89 Microcrystalline
cellulose (low moisture) Filler 147.11 Total capsule content
weight, mg 150.00 Size 2 Swedish orange gelatin capsule
Example 35
Pharmaceutical Composition 8
[0541] The composition is shown in Table 14 below.
TABLE-US-00014 TABLE 14 Batch composition Item g/ mg/ Number
Component Batch Capsule 1 Compound of formula (I-1) Form 2 7.06
0.30 2 Microcrystalline cellulose, NF 100 4.25 (Emcocel .RTM.
XLM90; low moisture) 3 Microcrystalline cellulose, NF 192.9 8.20
(Emcocel .RTM. XLM90; low moisture) 4 Microcrystalline cellulose,
NF 300 12.75 (Emcocel .RTM. XLM90; low moisture) 5 Microcrystalline
cellulose, NF 500 21.25 (Emcocel .RTM. XLM90; low moisture) 6
Microcrystalline cellulose, NF 900 38.25 (Emcocel .RTM. XLM90; low
moisture) Total weight 2000.0 85.00 Size 4 white opaque gelatin
capsules
[0542] The batch was prepared according to the following process:
[0543] 1) Microcrystalline cellulose, NF (Emcocel.RTM. XLM90; low
moisture) (Item #2) was screened through a 40 micron mesh screen.
[0544] 2) The screened material from step 1) was added to the PK
blender and was blended for 2 minutes. [0545] 3) Compound of
formula (I-1) Form 2 that had been screened through a 60 micron
mesh screen, was weighed out (Item #1). [0546] 4) The compound of
formula (I-1) Form 2 from step 3), and the microcrystalline
cellulose, NF (Emcocel.RTM. XLM90; low moisture) (Item #3) were
combined in a polyethylene bag, and the polyethylene bag was
shaken; then the contents of the polyethylene bag were passed
through the same 40 micron screen as was used in step 1). [0547] 5)
The material from step 4) was added to the PK blender and blended
for 15 minutes. [0548] 6) Microcrystalline cellulose, NF
(Emcocel.RTM. XLM90; low moisture) (Item #4) was screened through
the same 40 micron mesh screen, transferred to the same
polyethylene bag used in step 4) and shaken in the polyethylene
bag. [0549] 7) The material from step 6) was added to the PK
blender, which still contained the material from step 5) and
blended for 10 minutes. [0550] 8) Microcrystalline cellulose, NF
(Emcocel.RTM. XLM90; low moisture) (Item #5) was screened through
the same 40 micron mesh screen, transferred to the same
polyethylene bag used in steps 4) and 6), and shaken in the
polyethylene bag. [0551] 9) The material from step 8) was added to
the PK blender, which still contained the material from steps 5)
and 7), and blended for 10 minutes. [0552] 10) Microcrystalline
cellulose, NF (Emcocel.RTM. XLM90; low moisture) (Item #6) was
screened through the same 40 micron mesh screen, transferred to the
same polyethylene bag used in steps 4), 6), and 8), and shaken in
the polyethylene bag. [0553] 11) The material from step 10) was
added to the PK blender, which still contained the material from
steps 5), 7), and 9), and blended for 10 minutes. [0554] 12) The
material from the blender was encapsulated in size 4 white opaque
gelatin capsules using the In-Cap system. [0555] 13) The capsules
were de-dusted, and weight-sorted.
Example 36
Pharmaceutical Composition 9
[0556] The composition of the capsules is shown in Table 15
below.
TABLE-US-00015 TABLE 15 Capsule composition mg/ Component Function
Capsule Compound of formula (I-1) Form 2 0.3 Pregeletanized Starch
(Starch 1500) Filler 122.825 Talc Flow-aid 1.25 Magnesium Stearate
Lubricant 0.625 Total capsule content weight, mg 125.00 Size 4
white opaque gelatin capsule
Example 37
Pharmaceutical Composition 10
[0557] The composition of the capsules is shown in Table 16
below.
TABLE-US-00016 TABLE 16 Capsule composition mg/ Component Function
Capsule Compound of formula (I-1) Form 2 0.3 Pregeletanized Starch
(Starch 1500) Filler 124.7 Total capsule content weight, mg 125.00
Size 4 white opaque gelatin capsule
Example 38
Pharmaceutical Composition 11
[0558] The composition of the capsule is shown in Table 17
below.
TABLE-US-00017 TABLE 17 Capsule composition mg/ Component Function
Capsule Compound of formula (I-1) Form 2 0.3 Microcrystalline
cellulose Filler 124.7 (Emcocel .RTM. XLM90; low moisture) Talc
Flow-aid 1.25 Total capsule content weight, mg 125.00 Size 4 white
opaque gelatin capsule
Example 39
Pharmaceutical Composition 12
[0559] The composition of the capsule is shown in Table 18
below.
TABLE-US-00018 TABLE 18 Capsule composition mg/ Component Function
Capsule Compound of formula (I-1) Form 2 0.3 Microcrystalline
cellulose Filler 89.25 (Emcocel .RTM. XLM90; low moisture)
Magnesium Stearate Lubricant 0.45 Total capsule content weight, mg
90.00 Size 4 white opaque gelatin capsule
Example 40
Pharmaceutical Composition 13
[0560] The composition of the capsule is shown in Table 19
below.
TABLE-US-00019 TABLE 19 Capsule composition mg/ Component Function
Capsule Compound of formula (I-1) Form 2 0.3 Microcrystalline
cellulose Filler 88.35 (Emcocel .RTM. XLM90; low moisture) Talc
Flow-aid 0.9 Magnesium Stearate Lubricant 0.45 Total capsule
content weight, mg 90.00 Size 4 white opaque gelatin capsule
Example 41
Pharmaceutical Composition 14
[0561] The composition of the capsule is shown in Table 20
below.
TABLE-US-00020 TABLE 20 Capsule composition mg/ Component Function
Capsule Compound of formula (I-1) Form 2 0.3 Microcrystalline
cellulose Filler 51.15 (Emcocel .RTM. XLM90; low moisture) Talc
Flow-aid 0.98 Magnesium Stearate Lubricant 0.49 Pregeletanized
Starch (Starcap) 45.08 Total capsule content weight, mg 98.00 Size
4 white opaque gelatin capsule
Example 42
Pharmaceutical Composition 15
[0562] The composition of the capsule is shown in Table 21
below.
TABLE-US-00021 TABLE 21 Capsule composition mg/ Component Function
Capsule Compound of formula (I-1) Form 2 0.3 Microcrystalline
cellulose Filler 61.65 (Emcocel .RTM. XLM90; low moisture) Talc
Flow-aid 1.18 Magnesium Stearate Lubricant 0.59 Sodium Starch
Glycolate (Explotab) 54.28 Total capsule content weight, mg 118.00
Size 4 white opaque gelatin capsule
Example 43
Pharmaceutical Composition 16
[0563] The composition of the batch is shown in Table 22 below.
TABLE-US-00022 TABLE 22 Batch composition Item g/ mg/ Number
Component Batch Capsule 1 Compound of formula (I-1) Form 2 0.33
0.30 2 Microcrystalline cellulose, NF 5.00 4.50 (Emcocel .RTM.
XLM90; low moisture) 3 Microcrystalline cellulose, NF 8.17 7.35
(Emcocel .RTM. XLM90; low moisture) 4 Microcrystalline cellulose,
NF 14.00 12.60 (Emcocel .RTM. XLM90; low moisture) 5
Microcrystalline cellulose. NF 25.00 22.50 (Emcocel .RTM. XLM90;
low moisture) 6 Microcrystalline cellulose, NF 44.00 39.60 (Emcocel
.RTM. XLM90; low moisture) 7 Talc 1.00 0.90 8 Sodium Citrate 2.00
1.80 9 Magnesium Stearate 0.50 0.45 Total weight 100.00 90.00 Size
4 white opaque gelatin capsules
[0564] The batch was prepared according to the following process:
[0565] 1) Microcrystalline cellulose, NF (Emcocel.RTM. XLM90; low
moisture) (Item #2) was screened through a 40 micron mesh screen.
[0566] 2) The screened material from step 1) was added to the PK
blender and was blended for 2 minutes. [0567] 3) Compound of
formula (I-1) Form 2 that had been screened through a 60 micron
mesh screen, was weighed out (Item #1). [0568] 4) The compound of
formula (I-1) Form 2 from step 3), and the microcrystalline
cellulose, NF (Emcocel.RTM. XLM90; low moisture) (Item #3) were
combined in a polyethylene bag, and the polyethylene bag was
shaken; then the contents of the polyethylene bag were passed
through the same 40 micron screen as was used in step 1). [0569] 5)
The material from step 4) was added to the PK blender and blended
for 15 minutes. [0570] 6) Microcrystalline cellulose, NF
(Emcocel.RTM. XLM90; low moisture) (Item #4) was screened through
the same 40 micron mesh screen, transferred to the same
polyethylene bag used in step 4) and shaken in the polyethylene
bag. [0571] 7) Talc (Item #7), and Sodium Citrate (Item #8) were
screened through the same 40 micron mesh screen. [0572] 8) The
materials from step 6) and 7) were added to the PK blender, which
still contained the material from step 5) and blended for 10
minutes. [0573] 9) Microcrystalline cellulose, NF (Emcocel.RTM.
XLM90; low moisture) (Item #5) was screened through the same 40
micron mesh screen, transferred to the same polyethylene bag used
in steps 4) and 6), and shaken in the polyethylene bag. [0574] 10)
The material from step 9) was added to the PK blender, which still
contained the material from steps 5) and 8), and blended for 10
minutes. [0575] 11) Microcrystalline cellulose, NF (Emcocel.RTM.
XLM90; low moisture) (Item #6) was screened through the same 40
micron mesh screen, transferred to the same polyethylene bag used
in steps 4), 6), and 9), and shaken in the polyethylene bag. [0576]
12) The material from step 11) was added to the PK blender, which
still contained the material from steps 5), 8), and 10), and
blended for 10 minutes. [0577] 13) Magnesium Stearate (Item #9) was
screened through the same 40 micron mesh screen. [0578] 14) The
material from step 13) was added to the PK blender, which still
contained the material from steps 5), 8), 10), and 12) and blended
for 5 minutes. [0579] 15) The material from the blender was
encapsulated in size 4 white opaque gelatin capsules using the
Profill system. [0580] 16) The capsules were de-dusted, and
weight-sorted.
Example 44
Pharmaceutical Composition 17
[0581] The composition of the batch is shown in Table 23 below.
TABLE-US-00023 TABLE 23 Batch composition Item g/ mg/ Number
Component Batch Capsule 1 Compound of formula (I-1) Form 2 7.06
0.30 2 Microcrystalline cellulose, NF 4.94 0.21 (Emcocel .RTM.
XLM90; low moisture) 3 Microcrystalline cellulose, NF 25.00 1.06
(Emcocel .RTM. XLM90; low moisture) 4 Microcrystalline cellulose,
NF 53.00 2.25 (Emcocel .RTM. XLM90; low moisture) 5 Talc 10.00 0.43
6 Microcrystalline cellulose, NF 90 3.83 (Emcocel .RTM. XLM90; low
moisture) 7 Talc 30 1.28 8 Microcrystalline cellulose, 170 7.23 NF
(Emcocel .RTM. XLM90; low moisture) 9 Microcrystalline cellulose,
300 12.75 NF (Emcocel .RTM. XLM90; low moisture) 10
Microcrystalline cellulose, 500 21.25 NF (Emcocel .RTM. XLM90; low
moisture) 11 Microcrystalline cellulose, NF 800 34 (Emcocel .RTM.
XLM90; low moisture) 12 Magnesium Stearate 10 0.43 Total weight
2000 85 Size 4 white opaque gelatin capsules
[0582] The batch was prepared according to the following process:
[0583] 1) Microcrystalline cellulose, NF (Emcocel.RTM. XLM90; low
moisture) (Item #2) was screened through a 40 micron mesh screen.
[0584] 2) The screened material from step 1) was added to the small
PK blender and blended for 2 minutes. [0585] 3) Compound of formula
(I-1) Form 2 that had been screened through a 60 micron mesh screen
was weighed out (Item #1). [0586] 4) The compound of formula (I-1)
Form 2 from step 3), and the microcrystalline cellulose, NF
(Emcocel.RTM. XLM90; low moisture) (Item #3) were combined, and
then passed through the same 40 micron screen as was used in step
1). [0587] 5) The material from step 4) was added to the small PK
blender and blended for 30 minutes. [0588] 6) Microcrystalline
cellulose, NF (Emcocel.RTM. XLM90; low moisture) (Item #4) and Talc
(Item #5) were screened through the same 40 micron mesh screen.
[0589] 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. [0590] 8) Microcrystalline cellulose, NF
(Emcocel.RTM. XLM90; low moisture) (Item #6) was screened through
the same 40 micron mesh screen, transferred to a second larger PK
blender, and blended for 2 minutes. [0591] 9) The contents of the
small PK blender from steps 5) and 7) were emptied into a
polyethylene bag, and then transferred to the larger PK blender
from step 8). [0592] 10) Talc (Item #7) and Microcrystalline
cellulose, NF (Emcocel.RTM. XLM90; low moisture) (Item #8) were
screened through the same 40 micron mesh screen. [0593] 11) Half
the material from step 10) was added to the small PK blender 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. [0594] 12) The material from step 11) was added to the larger
PK blender, which still contained the material from steps 8) and
9). [0595] 13) The second half of material from step 10) was added
to the small PK blender from steps 5) and 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. [0596] 14) The
material from step 13) was added to the larger PK blender, which
still contained the material from steps 8), 9), and 12), and
blended for 10 minutes. [0597] 15) Microcrystalline cellulose, NF
(Emcocel.RTM. XLM90; low moisture) (Item #9) was screened through
the same 40 micron mesh screen, transferred to the same
polyethylene bag used in steps 9), 11), and 13), and shaken in the
polyethylene bag. [0598] 16) The material from step 15) was added
to the same larger PK blender, which still contained material from
steps 8), 9), 12), and 14), and blended for 10 minutes. [0599] 17)
Microcrystalline cellulose, NF (Emcocel.RTM. XLM90; low moisture)
(Item #10) was screened through the same 40 micron mesh screen.
[0600] 18) The material from step 17) was added to the same larger
PK blender, which still contained material from steps 8), 9), 12),
14), and 16), and blended for 10 minutes. [0601] 19)
Microcrystalline cellulose, NF (Emcocel.RTM. XLM90; low moisture)
(Item #11) was screened through the same 40 micron mesh screen.
[0602] 20) The material from step 19) was added to the same larger
PK blender, which still contained material from steps 8), 9), 12),
14), 16), and 18), and blended for 10 minutes. [0603] 21) Magnesium
Stearate (Item #12) was screened through the same 40 micron mesh
screen. [0604] 22) The material from step 21) was added to the same
larger PK blender, which still contained material from steps 8),
9), 12), 14), 16), 18), and 20), and blended for 5 minutes. [0605]
23) The material from the blender was encapsulated in size 4 white
opaque gelatin capsules using the Incap system. [0606] 24) The
capsules were de-dusted, and weight-sorted.
Example 45
Pharmaceutical Composition 18
[0607] The composition of the batch is shown in Table 24 below.
TABLE-US-00024 TABLE 24 Batch composition Item g/ mg/ Number
Component Batch Capsule 1 Compound of formula (I-1) Form 2 3.53
0.30 2 Microcrystalline cellulose, NF 496.5 4.50 (Emcocel .RTM.
XLM90; low moisture) 3 Microcrystalline cellulose, NF 500 7.35
(Emcocel .RTM. XLM90; low moisture) Total weight 1000 85 Size 4
white opaque gelatin capsules
[0608] The batch is prepared according to the following process:
[0609] 1) Microcrystalline cellulose, NF (Emcocel.RTM. XLM90; low
moisture) (Item #2) is screened through a 40 micron mesh screen and
added to a high shear mixer. [0610] 2) Compound of formula (I-1)
Form 2 is screened through a 60 micron mesh screen and weighed out
(Item #1) and added to the same high shear mixer from step 1).
[0611] 3) Microcrystalline cellulose, NF (Emcocel.RTM. XLM90; low
moisture) (Item #3) is screened through the same 40 micron mesh
screen, and added to the same high shear mixer from steps 1) and
2). [0612] 4) The high shear mixer from steps 1), 2), and 3) is run
for 4 minutes. [0613] 5) The material from the high shear mixer is
encapsulated in size 4 white opaque gelatin capsules using the
Incap system. [0614] 6) The capsules are de-dusted, and
weight-sorted.
Example 46
Lyophilized Powder 1
[0615] In a clean container, a solution of 40% tert-butyl
alcohol/60% water for injection was prepared by warming the
required amount of tert-butyl alcohol to 35.degree. C. and adding
water for injection. The solution was cooled to 15-30.degree. C. A
portion of the required amount (60% of the total batch) of
tert-butyl alcohol/water solution was added to a pre-compounding
container. Approximately 40% of the solution was reserved for use
in rinsing. Citric acid (30% of the batch amount) was added to the
pre-compounding container with stirring. The container was rinsed
with the reserved tert-butyl alcohol/water solution, and the rinses
were added to the pre-compounding container. The mixture was
stirred until the citric acid was completely dissolved. Sodium
citrate (30% of the batch amount) was added to the pre-compounding
container with stirring. The container was rinsed with the reserved
tert-butyl alcohol/water solution, and the rinses were added to the
pre-compounding container. The mixture was stirred until the sodium
citrate was completely dissolved.
N-(2-pyrazine)carbonyl-L-phenyl-L-leucine boronic acid (VIII-15)
was added to the pre-compouding container with stirring. The
container was rinsed with the reserved tert-butyl alcohol/water
solution, and the rinses were added to the pre-compounding
container. The mixture was stirred until the boronic acid was
completely dissolved. The citric acid, sodium citrate, and boronic
acid mixture from the pre-compounding container was transferred to
the main compounding vessel. The pre-compounding container was
rinsed with water for injection and the rinses were added to the
main compounding vessel. Citric acid (70% of the batch amount) was
added to the main vessel with stirring. The container was rinsed
with the water, and the rinses were added to the main vessel. The
mixture was stirred until the citric acid was completely dissolved.
Sodium citrate (70% of the batch amount) was added to the main
vessel with stirring. The container was rinsed with water, and the
rinses were added to the pre-compounding container. The mixture was
stirred until the sodium citrate was completely dissolved. Glycine
was added to the main vessel and the residual glycine was rinsed
with water, and the rinses were added to the main vessel. 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 thrugh a 0.22 .mu.m filter.
Aliquots of the filtered solution were placed into vials. The vials
were sealed with lyophilization stoppers and were placed on
lyophilizer chamber shelves maintained at 20.degree. C. The
lyophilization chamber shelves were cooled to -45.degree. C. using
the appropriate ramp rate and held at that temperature for 200
minutes. The shelf was warmed to -20.degree. C. using an
appropriate ramp rate and maintained at that temperature for 480
minutes. The shelf was re-cooled to -45.degree. C. using an
appropriate ramp rate and maintained at that temperature. After 200
minutes, the lyophililization chamber was evacuated, and the
chamber pressure was adjusted to 150 microns with nitrogen. The
chamber shelves were warmed up to -25.degree. C. using an
appropriate ramp rate, and held at that temperature for 3000
minutes. After each of the product thermocouples read -25.degree.
C. or warmer, the shelf was warmed to 27.degree. C. and maintained
at that temperature for 600 minutes. At the end of the terminal
drying phase, the chamber pressure was restored using nitrogen, and
vials were sealed and removed. Pre lyophilized solution contained:
52 mM citrate, 3% glycine, 4.7% tert-butyl alcohol (as shown in
Table 25 below).
TABLE-US-00025 TABLE 25 Batch composition Amount/ Amount No.
Component mL mM Batch per vial 1. Compound 0.001 g 2.6 0.300 g 3.5
mg (VIII-15) 2. Citric Acid 0.00382 g 18.2 1.147 g 13.37 mg
Monohydrate, USP/EP 3. Sodium Citrate 0.00994 g 33.8 2.982 g 34.79
mg Dihydrate, USP/EP 4. Glycine, 0.03 g 399.6 9.0 g 105 mg USP/EP
5. Tert-butyl n/a n/a 14.1 mL 0.1645 mL alcohol, ACS grade 6. Water
for n/a n/a Fill to batch n/a Injection, volume USP/EP 7. Total
volume n/a n/a 300 mL 3.5 mL 8. Final measured n/a n/a 5.08 n/a
pH
Example 47
Lyophilized Powder 2
[0616] Prepared as described in Example 46. Pre-lyophilized
solution contained: 52 mM citrate; 3% glycine; and 4.7% tert-butyl
alcohol (as shown in Table 26 below).
TABLE-US-00026 TABLE 26 Batch composition Amount/ Amount No.
Component mL mM Batch per vial 1. Compound 0.001 g 2.6 0.300 g 3.5
mg (VIII-15) 2. Citric Acid 0.00168 g 8.0 0.504 g 5.88 mg
Monohydrate, USP/EP 3. Sodium Citrate 0.0129 g 44.0 3.882 g 45.15
mg Dihydrate, USP/EP 4. Glycine, 0.03 g 399.6 9.0 g 105 mg USP/EP
5. Tert-butyl n/a n/a 14.1 mL 0.1645 mL alcohol, ACS grade 6. Water
for n/a n/a Fill to batch n/a Injection, volume USP/EP 7. Total
volume n/a n/a 300 mL 3.5 mL 8. Final measured n/a n/a 5.84 n/a
pH
Example 48
Lyophilized Powder 3
[0617] The formulation was prepared as described in Example 46,
except that the lyophilization cycle was modified. The vials were
sealed with lyophilization stoppers and placed on lyophilizer
chamber shelves maintained at 20.degree. C. The lyophilization
chamber shelves were cooled to -45.degree. C. using the appropriate
ramp rate and held at that temperature for 200 minutes. The shelf
was warmed to -20.degree. C. using an appropriate ramp rate and
maintained at that temperature for 480 minutes. The shelf was
re-cooled to -45.degree. 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 with nitrogen. The chamber shelves was
warmed up to -15.degree. C. using an appropriate ramp rate, and
held at that temperature for 2700 minutes. After each of the
product thermocouples read -15.degree. C. or warmer, the shelf was
warmed to 37.degree. C. and maintained at that temperature for 300
minutes. At the end of the terminal drying phase, the chamber
pressure was restored using nitrogen, and the vials were sealed and
removed. Pre-lyophilized solution contained: 52 mM citrate; 3%
glycine; and 4.7% tert-butyl alcohol (as shown in Table 27
below).
TABLE-US-00027 TABLE 27 Batch composition Amount/ Amount No.
Component mL mM Batch per vial 1. Compound 0.001 g 2.6 1.0 g 3.5 mg
(VIII-15) 2. Citric Acid 0.00382 g 18.2 3.82 g 13.37 mg
Monohydrate, USP/EP 3. Sodium Citrate 0.00994 g 33.8 9.94 g 34.79
mg Dihydrate, USP/EP 4. Glycine, 0.03 g 399.6 30.0 g 105 mg USP/EP
5. Tert-butyl n/a n/a 47.0 mL 0.1645 mL alcohol, ACS grade 6. Water
for n/a n/a Fill to batch n/a Injection, volume USP/EP 7. Total
volume n/a n/a 1000 mL 3.5 mL 8. Final measured n/a n/a 5.05 n/a
pH
Example 49
Lyophilized Powder 4
[0618] A clean vessel was charged 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-leucine boronic
acid (VIII-15) was added and stirred until dissolved. Glycine was
added to the vessel and the residual glycine was rinsed with water,
and the rinses were added to the main vessel. The mixture was
stirred until the glycine was completely dissolved. Sufficient
water was added to batch volume. The mixture was filtered through a
0.22 um filter. Aliquots of the filtered solution were placed into
vials. The vials were sealed with lyophilization stoppers and were
placed on lyophilizer chamber shelves maintained at 20.degree. C.
The lyophilization chamber shelves were cooled to -45.degree. C.
using the appropriate ramp rate and held at that temperature for
200 minutes. The shelf was warmed to -20.degree. C. using an
appropriate ramp rate and maintained at that temperature for 480
minutes. The shelf was re-cooled to -45.degree. C. using an
appropriate ramp rate and maintained at that temperature. After 200
minutes, the lyophilization chamber was evacuated with and the
chamber pressure was adjusted to 150 microns with nitrogen. The
chamber shelves were warmed up to -25.degree. C. using an
appropriate ramp rate, and held at that temperature for 3000
minutes. After each of the product thermocouples read -25.degree.
C. or warmer, the shelf was warmed to 27.degree. C. and maintained
at that temperature for 600 minutes. At the end of the terminal
drying phase, the chamber pressure was restored using nitrogen, and
vials were sealed and removed. Pre-lyophilized solution contained:
52 mM citrate; and 3% glycine (as shown in Table 28 below).
TABLE-US-00028 TABLE 28 Batch composition Amount/ Amount Component
mL mM Batch per vial 1. Compound 0.001 g 2.6 0.30 g 3.5 mg
(VIII-15) 2. Citric Acid 0.004097 g 19.5 1.229 g 14.34 mg
Monohydrate, USP/EP 3. Sodium Citrate 0.009557 g 32.5 2.867 g 33.45
mg Dihydrate, USP/EP 4. Glycine, USP/EP 0.03 g 399.6 9.0 g 105 mg
5. Water for Injection, n/a n/a Fill to batch n/a USP/EP volume 6.
Total volume n/a n/a 300 mL 3.5 mL 7. Final measured n/a n/a 4.90
n/a pH
Example 50
Lyophilized Powder 5
[0619] Prepared as described in Example 49. Pre-lyophilized
solution contained: 52 mM citrate; and 3% glycine (as shown in
Table 29 below). In this example, the pH of the pre-lyophilized
solution was adjusted to the final measured pH by the addition of
2N HCl.
TABLE-US-00029 TABLE 29 Batch composition Amount/ Amount No.
Component mL mM Batch per vial 1. Compound 0.001 g 2.6 0.30 g 3.5
mg (VIII-15) 2. Citric Acid 0.00168 g 8.0 0.504 g 5.88 mg
Monohydrate, USP/EP 3. Sodium Citrate 0.01294 g 44.0 3.882 g 45.29
mg Dihydrate, USP/EP 4. Glycine, 0.03 g 399.6 9.0 g 105 mg USP/EP
5. Water for Injection, n/a n/a Fill to batch n/a USP/EP volume 6.
Total batch volume n/a n/a 300 mL 3.5 mL 7. Final measured n/a n/a
5.84 n/a pH
Example 50
Lyophilized Powder 6
[0620] A clean vessel is charged with water for injection. Citric
acid and sodium citrate are added and stirred until dissolved. To
this solution,
4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-m-
ethylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid (I-1) is
added and stirred until dissolved. Glycine is added to the vessel
and the residual glycine is rinsed with water, and the rinses are
added to the main vessel. The mixture is stirred until the glycine
is completely dissolved. Sufficient water is added to batch volume.
The mixture is filtered through a 0.22 .mu.m filter. Aliquots of
the filtered solution are placed into sterilized vials. The vials
are sealed with lyophilization stoppers and are placed on
lyophilizer chamber shelves maintained at 20.degree. C. The
lyophilization chamber shelves are cooled to -45.degree. C. using
an appropriate ramp rate, and then are held at that temperature for
200 minutes. The shelf is warmed to -20.degree. C. using an
appropriate ramp rate and then is maintained at that temperature
for 480 minutes. The shelf is re-cooled to -45.degree. C. using an
appropriate ramp rate and is maintained at that temperature. After
200 minutes, the lyophilization chamber is evacuated, and the
chamber pressure is adjusted to 150 microns with nitrogen. The
chamber shelves are warmed up to -25.degree. C. using an
appropriate ramp rate, and held at that temperature for 3000
minutes. After each of the product thermocouples reads -25.degree.
C. or warmer, the shelf is warmed to 27.degree. C. and is
maintained at that temperature for 600 minutes. At the end of the
terminal drying phase, the chamber pressure is restored using
nitrogen, and vials are sealed and removed. The composition of the
pre-lyophilized solution is 55 mM citrate; and 3% glycine (as shown
in Table 30 below).
TABLE-US-00030 TABLE 30 Batch composition Amount/ Amount No.
Component mL mM Batch per vial 1. Compound 0.001 g 2.75 0.50 g 3.5
mg (I-1) [expressed as amount of compound (VIII-1)] 2. Citric Acid
0.0012 g 5.5 0.578 g 4.2 mg Monohydrate, USP/EP 3. Sodium Citrate
0.0147 g 49.5 7.279 g 51.45 mg Dihydrate, USP/EP 4. Glycine, 0.03 g
399.6 15.0 g 105 mg USP/EP 5. Water for n/a n/a Fill to batch n/a
Injection, volume USP/EP 6. Total volume n/a n/a 500 mL 3.5 mL
Example 51
Reconstitution of Lyophilized Powders
[0621] The lyophilized powders (e.g. as prepared in Examples 46-50)
are analyzed using XRPD, DSC, gas chromatography, and Karl Fisher
for cake structure, cake stability, residual solvent, and residual
moisture, respectively. The lyophilized powders are reconstituted
with the appropriate amount of sterile water for injection or with
sterile 0.9% sodium chloride solution for injection. The
reconstituted solutions are analyzed using HPLC, and NMR, for
purity, and percentage ester.
Example 52
Preparation of formulation of
4-(R,S)-(carboxymethyl)-2-((R)-1-(2-(2,5-dichlorobenzamido)acetamido)-3-m-
ethylbutyl)-6-oxo-1,3,2-dioxaborinane-4-carboxylic acid (I-1) Form
2 for parenteral or oral administration
[0622] A vessel was charged 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)acet-
amido)-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 batch
volume and the solution was filtered through a 0.2 .mu.m PES
membrane. Aliquots of the filtered solution were placed into vials.
The vials were sealed with stoppers and stored at -20.degree. C.
The batch and vial composition is as described below in Table
31.
TABLE-US-00031 TABLE 31 Batch composition Amount/ Amount No.
Component mL mM Batch per vial 1. Compound 0.001 g 2.75 20 g 3.3 mg
(I-1) [expressed as amount of compound (VIII-1)] 2. Citric Acid
0.0012 g 5.5 23.198 g 3.282 mg Monohydrate, USP/EP 3. Sodium
Citrate 0.0147 g 49.5 291.183 g 48.05 mg Dihydrate, USP/EP 4.
Sodium 0.0045 g 77 89.991 g 14.85 mg Chloride, USP/EP 5. Water for
n/a n/a Fill to batch n/a Injection, volume USP/EP 6. Total volume
n/a n/a 20 L 3.3 mL 7. Final measured n/a n/a 5.72 n/a pH
Example 53
Analytical Test Method 1
[0623] Reversed-phase HPLC using a C8 column at 25.degree. C. with
ultraviolet (UV) detection at 225 nm.
[0624] Mobile phase: The gradient system starts at 85% mobile phase
A (0.01% trifluoroacetic acid in water) and 15% mobile phase B
(0.01% trifluoroacetic acid in acetonitrile) and ends at 75% mobile
phase B after 40 minutes.
[0625] The test sample is prepared by dissolving the content of the
capsules in diluent which is 15:85 (v/v) acetonitrile:20mM citrate
buffer. Under these aqueous conditions, the compound of formula
(I-1) completely hydrolyzes the citrate ester portion of the
molecule to give the compound of formula (VIII-1) in a 1:1
molecular ratio. The presence of the compound of formula (VIII-I)
in the test sample is confirmed by comparison of the sample
retention time to that of the reference standard. The amount of the
compound of formula (VIII-1) present in a sample is calculated from
the area under the peak, on a weight-to-weight comparison including
molecular weight conversion, with the area under the peak of the
reference standard. The reference standard employed is a known
amount of the compound of formula (I-1), of known purity, which is
prepared under the same hydrolyzing conditions as the test sample.
The limit of quantitation for the method is 0.05% and the
calculated limit of detection is 0.02%.
Example 54
Analytical Test Method 2
[0626] 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
at 25.degree. C. for 8 minutes, with UV detection at 230 nm.
[0627] The test sample is prepared by dissolving the content of the
capsules in 40/60 (v/v) THF/n-Hexane. 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, on a weight to weight comparison, with the area under the
peak of the reference standard. The reference standard employed is
a known amount of the compound of formula (VIII-1), of known
purity, which is prepared under the same conditions as the test
sample. The limit of quantitation for detection of the compound of
formula (I-1) is 0.2%.
[0628] To calculate the amount of compound of formula (I-1) present
in a test sample, both Analytical Test Method 1 and Analytical Test
Method 2 are used. Analytical Test Method 1 is used to calculate
the amount on a weight basis of the compound of formula (VIII-1)
that is present in a test sample, containing the compound of
formula (I-1). Analytical Test Method 2 is also used to calculate
the amount of the compound of formula (VIII-1) present in the
sample of the compound of formula (I-1) obtained without induced
hydrolysis.
[0629] The amount of the compound of formula (VIII-1) obtained from
Analytical Test Method 1 minus the amount of the compound of
formula (VIII-1) obtained from Analytical Test Method 2 gives the
measured amount of the compound of formula (VIII-1) that is
produced by the induced hydrolysis of the compound of formula (I-1)
present in the test sample. Based on a 1:1 molecular ratio, a
molecular weight calculation gives the amount of the compound of
formula (I-1) present in the test sample.
[0630] While the foregoing invention has been described in some
detail for purposes of clarity and understanding, these particular
embodiments are to be considered as illustrative and not
restrictive. It will be appreciated by one skilled in the art from
a reading of this disclosure that various changes in form and
detail can 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.
[0631] The patent and scientific literature referred to herein
establishes knowledge that is available to those with skill in the
art. Unless otherwise defined, all technical and scientific terms
used herein have the same meaning as commonly understood by one of
ordinary skill in the art to which this invention belongs. The
issued patents, applications, and references that are cited herein
are hereby incorporated by reference to the same extent as if each
was specifically and individually indicated to be incorporated by
reference. In the case of inconsistencies, the present disclosure,
including definitions, will control.
* * * * *