U.S. patent application number 16/550967 was filed with the patent office on 2019-12-19 for hypdh inhibitors and methods of use for the treatment of kidney stones.
The applicant listed for this patent is UAB Research Foundation, Wake Forest University Health Sciences. Invention is credited to Ross P. Holmes, W. Todd Lowther, Daniel Yohannes.
Application Number | 20190382336 16/550967 |
Document ID | / |
Family ID | 56544199 |
Filed Date | 2019-12-19 |
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United States Patent
Application |
20190382336 |
Kind Code |
A1 |
Lowther; W. Todd ; et
al. |
December 19, 2019 |
HYPDH Inhibitors and Methods of Use for the Treatment of Kidney
Stones
Abstract
Provided herein are compounds of Formula I, Formula II, and
Formula III, and compositions comprising the same, as well as
methods of use thereof for controlling or inhibiting the formation
of calcium oxalate kidney stones, inhibiting the production of
glyoxylate and/or oxalate, and/or inhibiting hydroxyproline
dehydrogenase (HYPDH).
Inventors: |
Lowther; W. Todd;
(Pfafftown, NC) ; Holmes; Ross P.; (Birmingham,
AL) ; Yohannes; Daniel; (Winston-Salem, NC) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Wake Forest University Health Sciences
UAB Research Foundation |
Winston-Salem
Birmingham |
NC
AL |
US
US |
|
|
Family ID: |
56544199 |
Appl. No.: |
16/550967 |
Filed: |
August 26, 2019 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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15545818 |
Jul 24, 2017 |
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PCT/US2016/014707 |
Jan 25, 2016 |
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16550967 |
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62107701 |
Jan 26, 2015 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C07D 307/24 20130101;
C07C 62/24 20130101; C07C 59/11 20130101; C07C 229/48 20130101;
C07C 2601/08 20170501; C07D 333/38 20130101; C07D 307/68 20130101;
C07C 62/02 20130101; C07D 233/90 20130101 |
International
Class: |
C07C 229/48 20060101
C07C229/48; C07C 59/11 20060101 C07C059/11; C07C 62/24 20060101
C07C062/24; C07C 62/02 20060101 C07C062/02; C07D 307/68 20060101
C07D307/68; C07D 307/24 20060101 C07D307/24; C07D 233/90 20060101
C07D233/90; C07D 333/38 20060101 C07D333/38 |
Goverment Interests
GOVERNMENT FUNDING
[0002] This invention was made with government support under grant
numbers DK083527 and DK073732 awarded by National Institutes of
Health. The United States government has certain rights in the
invention.
Claims
1. A compound of Formula I, a compound of Formula II, or a compound
of Formula III: ##STR00014## wherein: X is O, S, NH, NMe or
CR.sup.xR.sup.y, wherein R.sup.xR.sup.y are each independently
selected from H, alkyl and halo; n is 0, 1, 2, 3, 4, 5 or 6; m is
0, 1, 2, or 3; R.sup.1 is selected from the group consisting of:
alkyl, alkenyl, alkynyl, aryl, halo, hydroxy, amine and carboxy;
R.sup.2 is selected from the group consisting of: H, alkyl,
hydroxy, amine, and .dbd.O; or R.sup.2 is R.sup.2aR.sup.2b, wherein
R.sup.2a and R.sup.2b are each independently selected from alkyl
and hydroxy; R.sup.3 is selected from the group consisting of: H,
hydroxy, amine, and .dbd.O; or R.sup.3 is R.sup.3aR.sup.3b, wherein
R.sup.3a and R.sup.3b are each independently selected from alkyl
and hydroxy; R.sup.4 is selected from the group consisting of: H,
alkyl, and hydroxy; or R.sup.4 is R.sup.4aR.sup.4b wherein R.sup.4a
and R.sup.4b are each independently selected from alkyl, hydroxy,
and halo, wherein said alkyl may be unsubstituted or substituted 1,
2 or 3 times with hydroxy; and each R.sup.5 is independently
selected from the group consisting of: H, alkyl, hydroxy, amine,
and .dbd.O; or R.sup.5 is R.sup.5aR.sup.5b wherein R.sup.5a and
R.sup.5b are each independently selected from alkyl and hydroxy; or
R.sup.2 and an adjacent R.sup.5 are taken together to form an aryl
or heteroaryl, or a pharmaceutically acceptable salt or prodrug
thereof.
2. The compound of claim 1, wherein said compound is a compound of
Formula I: ##STR00015## wherein: X is O, S, or CR.sup.xR.sup.y,
wherein R.sup.xR.sup.y are each H; and when X is S, the compound is
a compound of Formula I(A): ##STR00016## n is 0 or 1, and n is 1
when X is CR.sup.xR.sup.y and R.sup.3 is hydroxy; R.sup.1 is
carboxy; R.sup.2 is H; R.sup.3 is selected from the group
consisting of: hydroxy, amine, and .dbd.O; or R.sup.3 is
R.sup.3aR.sup.3b, wherein R.sup.3a and R.sup.3b are each
independently selected from alkyl and hydroxy; and R.sup.4 is H, or
a pharmaceutically acceptable salt thereof.
3. The compound of claim 1, wherein said compound is a compound of
Formula I: ##STR00017## wherein: X is CR.sup.xR.sup.y, wherein
R.sup.xR.sup.y are each H; n is 0 or 1, and n is 1 when R.sup.3 is
hydroxy; R.sup.1 is carboxy; R.sup.2 is H; R.sup.3 is selected from
the group consisting of: hydroxy, amine, and .dbd.O; or R.sup.3 is
R.sup.3aR.sup.3b, wherein R.sup.3a and R.sup.3b are each
independently selected from alkyl and hydroxy; and R.sup.4 is H, or
a pharmaceutically acceptable salt thereof.
4. The compound of claim 1, wherein said compound is a compound of
Formula I: ##STR00018## wherein: X is S; n is 0; R.sup.1 is
selected from the group consisting of: alkyl, alkenyl, alkynyl,
aryl, halo, hydroxy, amine and carboxy; R.sup.2 is selected from
the group consisting of: H and lower alkyl; R.sup.3 is selected
from the group consisting of: H, hydroxy, amine, and .dbd.O; or
R.sup.3 is R.sup.3aR.sup.3b, wherein R.sup.3a and R.sup.3b are each
independently selected from alkyl and hydroxy; R.sup.4 is selected
from the group consisting of: H and lower alkyl; or a
pharmaceutically acceptable salt or prodrug thereof.
5. The compound of claim 1, wherein said compound is a compound of
Formula I(A): ##STR00019## wherein: R.sup.1 is selected from the
group consisting of: alkyl, alkenyl, alkynyl, aryl, halo, hydroxy,
amine and carboxy; and R.sup.3 is selected from the group
consisting of: H, hydroxy, amine, and .dbd.O; or R.sup.3 is
R.sup.3aR.sup.3b, wherein R.sup.3a and R.sup.3b are each
independently selected from alkyl and hydroxy; or a
pharmaceutically acceptable salt or prodrug thereof.
6. The compound of claim 1, wherein said compound is a compound of
Formula I(A): ##STR00020## wherein: R.sup.1 is carboxy; and R.sup.3
is selected from the group consisting of: H, hydroxy, amine, and
.dbd.O; or R.sup.3 is R.sup.3aR.sup.3b, wherein R.sup.3a and
R.sup.3b are each independently selected from alkyl and hydroxy; or
a pharmaceutically acceptable salt thereof.
7. The compound of claim 1, wherein said compound is a compound of
Formula I(A): ##STR00021## wherein: R.sup.1 is carboxy; and R.sup.3
is selected from the group consisting of: H, hydroxyl and amine; or
R.sup.3 is R.sup.3aR.sup.3b, wherein R.sup.3a and R.sup.3b are each
independently selected from alkyl and hydroxy; or a
pharmaceutically acceptable salt thereof.
8. The compound of claim 7, wherein R.sup.3 is hydroxy or
R.sup.3aR.sup.3b, wherein R.sup.3aR.sup.3b are each hydroxy, or a
pharmaceutically acceptable salt thereof.
9. The compound of claim 1, wherein said compound is: ##STR00022##
or a pharmaceutically acceptable salt thereof.
10. The compound of claim 1, wherein said compound is selected from
the group consisting of: ##STR00023## and pharmaceutically
acceptable salts thereof.
11. The compound of claim 1, wherein said compound is selected from
the group consisting of: ##STR00024## and pharmaceutically
acceptable salts thereof.
12. A pharmaceutical composition comprising a compound,
pharmaceutically acceptable salt or prodrug of claim 1.
13. The composition of claim 12, wherein said composition is
formulated for oral administration.
14. The composition of claim 12, wherein said composition is a food
product formulation.
15. The composition of claim 12, wherein said composition is a
capsule, cachet, lozenge, or tablet.
16. The composition of claim 12, wherein said formulation is
provided in unit dosage form of from 1 mg to 10 grams of the
compound, pharmaceutically acceptable salt or prodrug.
Description
RELATED APPLICATIONS
[0001] This application is a divisional of U.S. patent application
Ser. No. 15/545,818, filed Jul. 24, 2017, which claims priority to
35 U.S.C. .sctn. 371 national phase entry of PCT Application
PCT/US2016/014707, filed Jan. 25, 2016, and published in English on
Aug. 4, 2016, as International Publication No. WO 2016/123012, and
claims the benefit of U.S. Provisional Patent Application Ser. No.
62/107,701, filed Jan. 26, 2015, the disclosure of each of which is
incorporated by reference herein in its entirety.
BACKGROUND
[0003] Kidney stones affect approximately 1 in 11 individuals in
the United States. The 2012 National Health and Nutrition and
Examination Survey (NHANES), part of the Urological Diseases in
America Project, reported that the overall prevalence of kidney
stones was 8.8% (10.6% and 7.1% for men and women, respectively)
(Jiang et al., Am J Physiol Gastrointest Liver Physiol 302,
G637-643, 2012). This study and others attest to the significant
increase in stone cases in general, but especially in individuals
with obesity, diabetes, and following bariatric surgery (Jiang et
al., supra; Knight et al., Am J Nephrol 25, 171-175, 2005). The
direct and indirect costs associated with kidney stone treatment
(i.e., nephrocalcinosis) are significant (Knight et al., Kidney Int
70, 1929-1934, 2006).
[0004] Individuals with Primary Hyperoxaluria (PH) have mutations
in a variety of genes involved in glyoxylate and hydroxyproline
(Hyp) metabolism that result in a significant increase in oxalate
production and deposition of calcium oxalate stones, the most
common type of stones for all stone formers. The treatments for
these individuals range from a combined kidney-liver transplant to
a life-long use of potassium citrate, increased fluid intake and
dietary restriction of oxalate (Riedel et al., PLoS One 6, e26021,
2011; Knight et al., Am J Physiol-Renal 302, F688-693, 2012).
Treatments for the removal of stones currently include shock-wave
lithotripsy, ureteroscopic stone removal, and percutaneous
nephrolithotomy (Riedel et al., supra). However, the recurrence of
stones following the available procedures is over 50%.
[0005] Kidney stones are also a significant problem in veterinary
medicine. Pets such as dogs and cats can develop stones that lead
to painful urination and/or a life-threatening blockage.
[0006] Considering that the current treatments only address
symptoms, novel treatments to prevent or control the formation of
stones in PH and other idiopathic stone formers are greatly
needed.
SUMMARY
[0007] Provided herein according to some embodiments is a method of
inhibiting the formation of oxalate kidney stones, comprising:
administering to a subject in need thereof a therapeutically
effective amount of a compound of Formula I, a compound of Formula
II, or a compound of Formula III:
##STR00001##
wherein:
[0008] X is O, S, NH, NMe or CR.sup.xR.sup.y, wherein R.sup.x and
R.sup.y are each independently selected from H, alkyl and halo;
[0009] n is 0, 1, 2, 3, 4, 5 or 6;
[0010] m is 0, 1, 2, or 3;
[0011] R.sup.1 is selected from the group consisting of: alkyl,
alkenyl, alkynyl, aryl, halo, hydroxy, amine and carboxy;
[0012] R.sup.2 is selected from the group consisting of: H, alkyl
(e.g., lower alkyl), hydroxy, amine, and .dbd.O; or R.sup.2 is
R.sup.2aR.sup.2b, wherein R.sup.2a and R.sup.2b are each
independently selected from alkyl (e.g., lower alkyl) and
hydroxy;
[0013] R.sup.3 is selected from the group consisting of: H,
hydroxy, amine, and .dbd.O; or R.sup.3 is R.sup.3aR.sup.3b, wherein
R.sup.3a and R.sup.3b are each independently selected from alkyl
(e.g., lower alkyl) and hydroxy;
[0014] R.sup.4 is selected from the group consisting of: H, alkyl
(e.g., lower alkyl), and hydroxy; or R.sup.4 is R.sup.4aR.sup.4b
wherein R.sup.4a and R.sup.4b are each independently selected from
alkyl (e.g., lower alkyl), hydroxy, and halo, wherein said alkyl
may be unsubstituted or substituted 1, 2 or 3 times with hydroxy;
and
[0015] each R.sup.5 is independently selected from the group
consisting of: H, alkyl (e.g., lower alkyl), hydroxy, amine, and
.dbd.O; or R.sup.5 is R.sup.5aR.sup.5b wherein R.sup.5a and
R.sup.5b are each independently selected from alkyl (e.g., lower
alkyl) and hydroxy; or R.sup.2 and an adjacent R.sup.5 are taken
together to form an aryl or heteroaryl, or a pharmaceutically
acceptable salt or prodrug thereof.
[0016] In some embodiments, the compound is a compound of Formula
I:
##STR00002##
wherein:
[0017] X is S;
[0018] n is 0;
[0019] R.sup.1 is selected from the group consisting of: alkyl,
alkenyl, alkynyl, aryl, halo, hydroxy, amine and carboxy;
[0020] R.sup.2 is selected from the group consisting of: H and
lower alkyl;
[0021] R.sup.3 is selected from the group consisting of: hydroxy,
amine, and .dbd.O; or R.sup.3 is R.sup.3aR.sup.3b, wherein R.sup.3a
and R.sup.3b are each independently hydroxy; and
[0022] R.sup.4 is selected from the group consisting of: H and
lower alkyl;
[0023] or a pharmaceutically acceptable salt or prodrug
thereof.
[0024] Also provided is a method of inhibiting the production of
glyoxylate and/or oxalate in a subject in need thereof, comprising:
administering to the subject a therapeutically effective amount of
a compound of Formula I, a compound of Formula II, or a compound of
Formula III:
##STR00003##
wherein:
[0025] X is O, S, NH, NMe or CR.sup.xR.sup.y, wherein R.sup.x and
R.sup.y are each independently selected from H, alkyl and halo;
[0026] n is 0, 1, 2, 3, 4, 5 or 6;
[0027] m is 0, 1, 2, or 3;
[0028] R.sup.1 is selected from the group consisting of: alkyl,
alkenyl, alkynyl, aryl, halo, hydroxy, amine and carboxy;
[0029] R.sup.2 is selected from the group consisting of: H, alkyl
(e.g., lower alkyl), hydroxy, amine, and .dbd.O; or R.sup.2 is
R.sup.2aR.sup.2b, wherein R.sup.2a and R.sup.2b are each
independently selected from alkyl (e.g., lower alkyl) and
hydroxy;
[0030] R.sup.3 is selected from the group consisting of: H,
hydroxy, amine, and .dbd.O; or R.sup.3 is R.sup.3aR.sup.3b, wherein
R.sup.3a and R.sup.3b are each independently selected from alkyl
(e.g., lower alkyl) and hydroxy;
[0031] R.sup.4 is selected from the group consisting of: H, alkyl
(e.g., lower alkyl), and hydroxy; or R.sup.4 is R.sup.4aR.sup.4b
wherein R.sup.4a and R.sup.4b are each independently selected from
alkyl (e.g., lower alkyl), hydroxy, and halo, wherein said alkyl
may be unsubstituted or substituted 1, 2 or 3 times with hydroxy;
and
[0032] each R.sup.5 is independently selected from the group
consisting of: H, alkyl (e.g., lower alkyl), hydroxy, amine, and
.dbd.O; or R.sup.5 is R.sup.5aR.sup.5b wherein R.sup.5a and
R.sup.5b are each independently selected from alkyl (e.g., lower
alkyl) and hydroxy; or R.sup.2 and an adjacent R.sup.5 are taken
together to form an aryl or heteroaryl,
[0033] or a pharmaceutically acceptable salt or prodrug
thereof.
[0034] In some embodiments, the compound is a compound of Formula
I:
##STR00004##
wherein:
[0035] X is S;
[0036] n is 0;
[0037] R.sup.1 is selected from the group consisting of: alkyl,
alkenyl, alkynyl, aryl, halo, hydroxy, amine and carboxy;
[0038] R.sup.2 is selected from the group consisting of: H and
lower alkyl;
[0039] R.sup.3 is selected from the group consisting of: hydroxy,
amine, and .dbd.O; or R.sup.3 is R.sup.3aR.sup.3b, wherein R.sup.3a
and R.sup.3b are each independently hydroxy; and
[0040] R.sup.4 is selected from the group consisting of: H and
lower alkyl;
[0041] or a pharmaceutically acceptable salt or prodrug
thereof.
[0042] Further provided is a method of inhibiting hydroxyproline
dehydrogenase (HYPDH) in a subject in need thereof, comprising:
administering to the subject a therapeutically effective amount of
a compound of Formula I, a compound of Formula II, or a compound of
Formula III:
##STR00005##
wherein:
[0043] X is O, S, NH, NMe or CR.sup.xR.sup.y, wherein R.sup.x and
R.sup.y are each independently selected from H, alkyl and halo;
[0044] n is 0, 1, 2, 3, 4, 5 or 6;
[0045] m is 0, 1, 2, or 3;
[0046] R.sup.1 is selected from the group consisting of: alkyl,
alkenyl, alkynyl, aryl, halo, hydroxy, amine and carboxy;
[0047] R.sup.2 is selected from the group consisting of: H, alkyl
(e.g., lower alkyl), hydroxy, amine, and .dbd.O; or R.sup.2 is
R.sup.2aR.sup.2b, wherein R.sup.2a and R.sup.2b are each
independently selected from alkyl (e.g., lower alkyl) and
hydroxy;
[0048] R.sup.3 is selected from the group consisting of: H,
hydroxy, amine, and .dbd.O; or R.sup.3 is R.sup.3aR.sup.3b, wherein
R.sup.3a and R.sup.3b are each independently selected from alkyl
(e.g., lower alkyl) and hydroxy;
[0049] R.sup.4 is selected from the group consisting of: H, alkyl
(e.g., lower alkyl), and hydroxy; or R.sup.4 is R.sup.4aR.sup.4b
wherein R.sup.4a and R.sup.4b are each independently selected from
alkyl (e.g., lower alkyl), hydroxy, and halo, wherein said alkyl
may be unsubstituted or substituted 1, 2 or 3 times with hydroxy;
and
[0050] each R.sup.5 is independently selected from the group
consisting of: H, alkyl (e.g., lower alkyl), hydroxy, amine, and
.dbd.O; or R.sup.5 is R.sup.5aR.sup.5b wherein R.sup.5a and
R.sup.5b are each independently selected from alkyl (e.g., lower
alkyl) and hydroxy; or R.sup.2 and an adjacent R.sup.5 are taken
together to form an aryl or heteroaryl,
[0051] or a pharmaceutically acceptable salt or prodrug
thereof.
[0052] In some embodiments, the compound is a compound of Formula
I:
##STR00006##
wherein:
[0053] X is S;
[0054] n is 0;
[0055] R.sup.1 is selected from the group consisting of: alkyl,
alkenyl, alkynyl, aryl, halo, hydroxy, amine and carboxy;
[0056] R.sup.2 is selected from the group consisting of: H and
lower alkyl;
[0057] R.sup.3 is selected from the group consisting of: hydroxy,
amine, and .dbd.O; or R.sup.3 is R.sup.3aR.sup.3b, wherein R.sup.3a
and R.sup.3b are each independently hydroxy; and
[0058] R.sup.4 is selected from the group consisting of: H and
lower alkyl;
[0059] or a pharmaceutically acceptable salt or prodrug
thereof.
[0060] Also provided is a compound of Formula I, a compound of
Formula II, or a compound of Formula III, or pharmaceutically
acceptable salt or prodrug thereof, as well as pharmaceutical
compositions comprising the same.
[0061] Further provided is the use of a compound of Formula I, a
compound of Formula II, or a compound of Formula III, or a
pharmaceutically acceptable salt or prodrug thereof, for
controlling or inhibiting the formation of calcium oxalate kidney
stones, inhibiting the production of glyoxylate and/or oxalate,
and/or inhibiting hydroxyproline dehydrogenase (HYPDH).
[0062] Also provided is the use of a compound of Formula I, a
compound of Formula II, or a compound of Formula III, or a
pharmaceutically acceptable salt or prodrug thereof, in the
preparation of a medicament for controlling or inhibiting the
formation of calcium oxalate kidney stones, inhibiting the
production of glyoxylate and/or oxalate, and/or inhibiting
hydroxyproline dehydrogenase (HYPDH).
BRIEF DESCRIPTION OF THE DRAWINGS
[0063] FIG. 1 presents a schematic of the metabolism of
4-hydroxyproline and glyoxylate within a hepatocyte. Four
mitochondrial enzymes are responsible for Hyp breakdown:
hydroxyproline dehydrogenase (HYPDH),
.DELTA..sup.1-pyrroline-5-carboxylate dehydrogenase (1P5CDH),
aspartate aminotransferase (AspAT), and 4-hydroxy-2-oxoglutarate
aldolase (HOGA). A variety of enzymes, including alanine-glyoxylate
aminotransferase (AGT), D-amino acid oxidase (DAO), glyoxylate
reductase (GR), and lactate dehydrogenase (LDH), can act on
glyoxylate produced from HOG cleavage. AGT, GR, and HOGA are
mutated within primary hyperoxaluria patients (type 1, 2, and 3,
respectively).
[0064] FIG. 2 presents the structures of Hyp analogs, of which some
have been tested for HYPDH inhibition.
DETAILED DESCRIPTION
[0065] Provided herein are methods of treatment for controlling or
inhibiting the formation of kidney stones comprising administering
to a subject in need thereof an inhibitor of hydroxyproline
dehydrogenase (HYPDH), as well as compounds and compositions useful
for the same.
[0066] The disclosures of all patent references cited herein are
hereby incorporated by reference to the extent they are consistent
with the disclosure set forth herein. As used herein in the
description of the invention and the appended claims, the singular
forms "a," "an" and "the" are intended to include the plural forms
as well, unless the context clearly indicates otherwise.
[0067] "Subject" or "patient" as used herein are generally
mammalian subjects, including both human subjects and non-human
mammalian subjects (e.g., dog, cat, horse, etc.) for research or
veterinary purposes. Subjects may be male or female and may be of
any suitable age, including neonate, infant, juvenile, adolescent,
adult, and geriatric subjects.
[0068] "Treat" as used herein refers to any type of treatment that
imparts a benefit to a subject, particularly slowing or inhibiting
the formation of glyoxylate and/or oxalate, slowing or inhibiting
the formation of calcium oxalate stones in the kidneys and/or
urinary tract (kidneys, ureters, bladder, and urethra), and/or the
deposition of calcium oxalate in other tissues such as the heart.
For example, the treatment may reduce the size of and/or decrease
the number of such stones, inhibit or slow the growth of such
stones or calcium oxalate deposition in tissues such as the heart,
alleviate symptoms of such stones or deposition, etc. Treatment
also includes prophylactic treatment of a subject deemed to be at
risk of kidney stone formation (e.g., after bariatric surgery).
[0069] "Kidney stones" are hard deposits of minerals that form a
stone or crystal aggregation, which may result in damage or failure
of the kidney and/or urinary tract function. Most kidney stones are
calcium stones, usually in the form of calcium oxalate.
[0070] "Oxalate" or "oxalic acid" is a dianion of the formula
C.sub.2O.sub.4.sup.2- produced by the body and also commonly
ingested in the diet. Oxalate can combine with calcium in the
kidneys or urinary tract to form calcium oxalate, which is the main
component of most kidney stones.
[0071] "Glyoxylate" is a precursor of oxalate, as shown in FIG.
1.
[0072] "Primary hyperoxaluria" is a condition characterized by the
overproduction of oxalate and/or defective production or function
of one or more enzymes that regulate the levels of oxalate in the
body. Sufferers of Type 1 primary hyperoxaluria have a defect or
shortage of the alanine:glyoxylate aminotransferase enzyme (AGT).
Type 2 primary hyperoxaluria sufferers have a defect or shortage of
the glyoxylate reductase enzyme (GR). Type 3 primary hyperoxaluria
sufferers have a defect or shortage of the 4-hydroxy-2-oxoglutarate
aldolase (HOGA).
[0073] "Hydroxyproline" or "Hyp" has the structure:
##STR00007##
Hydroxyproline is produced in the body primarily from endogenous
collagen turnover (Miyata et al., Proc Natl Acad Sci USA 111,
14406-14411, 2014). Using a unique metabolic tracer,
.sup.13C.sub.5,.sup.15N-Hyp (all five carbons isotope and nitrogen
atom labeled), it was determined that the level of Hyp turnover
could be as high as 6-7 g/day (Riedel et al., Biochim Biophys Acta
1822, 1544-1552, 2012). Less than 5 mg of free Hyp is excreted in
urine each day, indicating that most of the Hyp is metabolized
(Belostotsky et al., J Mol Med (Berl) 90, 1497-1504, 2012). This
significant metabolic load could contribute up to 25% of the
endogenous oxalate produced (Phang et al., (2001) Disorders of
proline and hydroxyproline metabolism. in The Metabolic and
Molecular Bases of Inherited Disease (Scriver, C. R., Beaudet, A.
L., Sly, W. S., Vallee, D., Childs, B., Kinzler, K. W., and
Vogelstein, B. eds.), McGraw-Hill, New York. pp 1821-1838). The
biological reason why Hyp metabolism occurs is not clear, although
it does enable some pyruvate to feed back into other pathways.
[0074] Hyp is metabolized primarily in the mitochondria of the
liver and renal cortical tissue (Kivirikko, Int Rev Connect Tissue
Res 5, 93-163, 1970; Atlante et al., Biochem Biophys Res Commun
202, 58-64, 1994; Monico et al., Clin J Am Soc Nepthrol 6,
2289-2295, 2011; Wold et al., J Food Sc 64, 377-383, 1999). Diet
can also be a source of collagen. For example, a quarter pound
hamburger rich in gristle could contain as much as 6 grams of
collagen, yielding 780 mg of Hyp (Khan et al., J Urol 184,
1189-1196, 2010). In fact, dietary Hyp can significantly increase
oxalate production in humans and lead to hyperoxaluria in mouse and
rat models (Khan et al., Kidney Int 70, 914-923, 2006; Valle et
al., J Clin Invest 64, 1365-1370, 1979; Adams et al., Annu Rev
Biochem 49, 1005-1061, 1980).
[0075] FIG. 1 presents the Hyp catabolic pathway, which involves
four enzymatic reactions (Miyata et al., Proc Natl Acad Sci USA
111, 14406-14411, 2014; Efron et al., New Engl J Med 272,
1299-1309, 1965; Pelkonen et al., New Engl J Med 283, 451-456,
1970). The first step of the pathway is the flavin FAD+-dependent
oxidation of Hyp to .DELTA..sup.1-pyrroline-3-hydroxy-5-carboxylate
(3-OH--P5C) by HYPDH. The 3-OH--P5C intermediate is converted to
4-hydroxy-glutamate (4-OH-Glu) by 1P5C dehydrogenase (1P5CDH), an
NAD+-dependent enzyme shared with the proline degradation pathway
(Efron et al., supra). Aspartate aminotransferase (AspAT) utilizes
oxaloacetate to convert 4-OH-Glu to 4-hydroxy-2-oxoglutarate (HOG).
HOG is then cleaved by the unique HOG aldolase (HOGA) into two
fragments, glyoxylate and pyruvate. The glyoxylate can then be
converted to glycolate and glycine via glyoxylate reductase (GR)
and alanine:glyoxylate aminotransferase (AGT), respectively.
[0076] AGT, GR, and HOGA are mutated within primary hyperoxaluria
patients (PH type 1, 2, and 3, respectively). For PH1 and PH2
patients, the glyoxylate produced from Hyp could exacerbate the
already high levels of glyoxylate, and increase oxalate production
via the lactate dehydrogenase (LDH). For PH3 patients, HOGA is
inactivated, leading to a buildup of HOG (Riedel et al., Biochim
Biophys Acta 1822, 1544-1552, 2012; Belostotsky et al., J Mol Med
(Berl) 90, 1497-1504, 2012). Recent studies identified that HOG can
inhibit GR, potentially leading to a PH2-like phenotype (Riedel et
al., Biochim Biophys Acta 1822, 1544-1552, 2012).
[0077] In contrast, hydroxyprolinemia, caused by deficiencies in
HYPDH, is not associated with any overt consequences, and Hyp is
safely excreted without being degraded (Curhan et al., Kidney Int
73, 489-496, 2008; Roy et al., Nature Protoc 5, 725-738, 2010).
[0078] Thus, and without wishing to be bound by theory, inhibition
of HYPDH by a small molecule inhibitor is not expected to lead to
any adverse side effects, and will block the formation of
glyoxylate and oxalate from Hyp for all PH patient types and the
buildup of HOG, 4-OH-Glu and dihydroxy-glutarate for PH3
patients.
[0079] Inhibition of HYPDH is also expected to help idiopathic
stone formers and other individuals with high urinary oxalate
levels, such as those that have undergone gastric bypass surgery.
For the latter, there is a significant increase in stone formation
that may benefit from prophylactic treatment post surgery. While
the exact origins of the oxalate in these patients has not been
determined, inhibition of HYPDH will decrease glyoxylate and
oxalate levels, which will ultimately reduce the glyoxylate and
oxalate burden in them.
1. Active Compounds.
[0080] As used herein in the accompanying chemical structures, "H"
refers to a hydrogen atom. "C" refers to a carbon atom. "N" refers
to a nitrogen atom. "O" refers to an oxygen atom. "Me" refers to a
methyl group.
[0081] The term "hydroxy," as used herein, refers to a group
--OH.
[0082] "Carbonyl" is a group having a carbon atom double-bonded to
an oxygen atom (C.dbd.O).
[0083] "Carboxy" as used herein refers to a group --COOH.
[0084] "Amine" or "amino" refers to a group --NH.sub.2.
[0085] "Halo" is a halogen group selected from the group consisting
of fluoro (--F), choro (--Cl), bromo (--Br), and iodo (--I).
[0086] "Alkyl," as used herein, refers to a saturated straight or
branched chain, or cyclic hydrocarbon containing from 1 to 10
carbon atoms. Representative examples of alkyl include, but are not
limited to, methyl, ethyl, n-propyl, iso-propyl, n-butyl,
sec-butyl, iso-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl,
n-hexyl, 3-methylhexyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl,
n-heptyl, n-octyl, n-nonyl, n-decyl, cyclopropyl, cyclobutyl,
cyclopentyl, cyclohexyl, and the like. "Lower alkyl" as used
herein, is a subset of alkyl and refers to a straight or branched
chain hydrocarbon group containing from 1 to 4 carbon atoms.
Representative examples of lower alkyl include, but are not limited
to, methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl,
tert-butyl, cyclopropyl, cyclobutyl, and the like.
[0087] "Alkenyl," as used herein, refers to a straight or branched
chain hydrocarbon containing from 2 to 10 carbons and containing at
least one carbon-carbon double bond formed by the removal of two
hydrogens. Representative examples of "alkenyl" include, but are
not limited to, ethenyl, 2-propenyl, 2-methyl-2-propenyl,
3-butenyl, 4-pentenyl, 5-hexenyl, 2-heptenyl, 2-methyl-1-heptenyl,
3-decenyl and the like. "Lower alkenyl" as used herein, is a subset
of alkenyl and refers to a straight or branched chain hydrocarbon
group containing from 2 to 4 carbon atoms and at least one
carbon-carbon double bond.
[0088] "Alkynyl," as used herein, refers to a straight or branched
chain hydrocarbon group containing from 2 to 10 carbon atoms and
containing at least one carbon-carbon triple bond. Representative
examples of alkynyl include, but are not limited, to acetylenyl,
1-propynyl, 2-propynyl, 3-butynyl, 2-pentynyl, 1-butynyl and the
like. "Lower alkynyl" as used herein, is a subset of alkynyl and
refers to a straight or branched chain hydrocarbon group containing
from 2 to 4 carbon atoms at least one carbon-carbon triple
bond.
[0089] "Aryl," as used herein, refers to a monocyclic carbocyclic
ring system or a bicyclic carbocyclic fused ring system having one
or more aromatic rings. Representative examples of aryl include the
monovalent species azulenyl, indanyl, indenyl, naphthyl, phenyl,
tetrahydronaphthyl, and the like, or divalent species thereof if
formed from R.sup.2 and an adjacent R.sup.5 taken together in
Formula II as taught herein.
[0090] "Heteroaryl," as used herein, refers to a monovalent
aromatic group having a single ring or two fused rings and
containing in the ring(s) at least one heteroatom (typically 1 to
3) selected from nitrogen, oxygen or sulfur. Representative
heteroaryl groups include, by way of example, monovalent species of
pyrrole, imidazole, thiazole, oxazole, furan, thiophene, triazole,
pyrazole, isoxazole, isothiazole, pyridine, pyrazine, pyridazine,
pyrimidine, triazine, indole, benzofuran, benzothiophene,
benzoimidazole, benzthiazole, quinoline, isoquinoline, quinazoline,
quinoxaline and the like, or divalent species thereof if formed
from R.sup.2 and an adjacent R.sup.5 taken together in Formula II
as taught herein.
[0091] As used herein, a specified group may be either
unsubstituted, or substituted by one or more suitable groups in
place of a hydrogen atom on the parent chain or cycle of an organic
molecule. For example, the alkyl, alkenyl, alkynyl, aryl and
heteroaryl groups of the invention may be unsubstituted or
substituted (e.g., 1, 2 or 3 times) with alkyl, hydroxy or
halo.
[0092] A "pharmaceutically acceptable salt" is a salt that retains
the biological effectiveness of the free acids or bases of a
specified compound and that is not biologically or otherwise
undesirable. Examples of pharmaceutically acceptable salts include
sulfates, pyrosulfates, bisulfates, sulfites, bisulfites,
phosphates, monohydrogenphosphates, dihydrogenphosphates,
metaphosphates, pyrophosphates, chlorides, bromides, iodides,
acetates, propionates, decanoates, caprylates, acrylates, formates,
isobutyrates, caproates, heptanoates, propiolates, oxalates,
malonates, succinates, suberates, sebacates, fumarates, maleates,
butyne-1,4-dioates, hexyne-1,6-dioates, benzoates, chlorobenzoates,
methylbenzoates, dinitrobenzoates, hydroxybenzoates,
methoxybenzoates, phthalates, sulfonates, xylenesulfonates,
phenylacetates, phenylpropionates, phenylbutyrates, citrates,
lactates, gamma-hydroxybutyrates, glycollates, tartrates,
methane-sulfonates, propanesulfonates, naphthalene-1-sulfonates,
naphthalene-2-sulfonates, and mandelates.
[0093] A "prodrug" is a compound that is converted under
physiological conditions or by solvolysis or metabolically to a
compound that is pharmaceutically active. A thorough discussion is
provided in T. Higuchi and V. Stella, Prodrugs as Novel delivery
Systems, Vol. 14 of the A.C.S. Symposium Series and in Edward B.
Roche, ed., Bioreversible Carriers in Drug Design, American
Pharmaceutical Association and Pergamon Press, 1987, both of which
are incorporated by reference herein in their entirety. See also
Huttunen et al., "Prodrugs--from Serendipity to Rational Design,"
Pharmacological Reviews 63(3):750-771 (2011), which is incorporated
by reference herein. Example prodrugs include, but are not limited
to, the addition of/conversion to phosphate(s), amino acid esters,
amino acid amides, sugar derivatives, alkyl or aryl esters, etc.,
at an --OH, --SH, --NH or --COOH group of the parent active
compound.
[0094] Provided herein as active compounds according to some
embodiments are compounds of Formula I:
##STR00008##
wherein:
[0095] X is O, S, NH, NMe or CR.sup.xR.sup.y, wherein R.sup.x and
R.sup.y are each independently selected from H, alkyl and halo;
[0096] n is 0, 1, 2, 3, 4, 5 or 6;
[0097] R.sup.1 is selected from the group consisting of: alkyl,
alkenyl, alkynyl, aryl, halo, hydroxy, amine and carboxy;
[0098] R.sup.2 is selected from the group consisting of: H, alkyl
(e.g., lower alkyl), hydroxy, amine, and .dbd.O; or R.sup.2 is
R.sup.2aR.sup.2b, wherein R.sup.2a and R.sup.2b are each
independently selected from alkyl (e.g., lower alkyl) and
hydroxy;
[0099] R.sup.3 is selected from the group consisting of: hydroxy,
amine, and .dbd.O; or R.sup.3 is R.sup.3aR.sup.3b, wherein R.sup.3a
and R.sup.3b are each independently hydroxy; and
[0100] R.sup.4 is selected from the group consisting of: H, alkyl
(e.g., lower alkyl), and hydroxy; or R.sup.4 is R.sup.4aR.sup.4b
wherein R.sup.4a and R.sup.4b are each independently selected from
alkyl (e.g., lower alkyl), hydroxy, and halo, wherein said alkyl
may be unsubstituted or substituted 1, 2 or 3 times with
hydroxy,
[0101] or a pharmaceutically acceptable salt or prodrug
thereof.
[0102] In some embodiments of Formula I, X is O, S, or
CR.sup.xR.sup.y.
[0103] In some embodiments of Formula I, n is 0 and/or R.sup.1 is
hydroxy.
[0104] In some embodiments of Formula I, R.sup.2 and/or R.sup.4 is
selected from the group consisting of: H and lower alkyl.
[0105] In some embodiments of Formula I, R.sup.3 is hydroxy.
[0106] In some embodiments, of Formula I, the compound is a
compound of Formula I(A):
##STR00009##
wherein:
[0107] R.sup.1 is selected from the group consisting of: alkyl,
alkenyl, alkynyl, aryl, halo, hydroxy, amine and carboxy; and
[0108] R.sup.3 is selected from the group consisting of: H,
hydroxy, amine, and .dbd.O; or R.sup.3 is R.sup.3aR.sup.3b, wherein
R.sup.3a and R.sup.3b are each independently selected from alkyl
(e.g., lower alkyl) and hydroxy; or a pharmaceutically acceptable
salt or prodrug thereof.
[0109] In some embodiments of Formula I(A), R.sup.1 is carboxy
and/or R.sup.3 is hydroxy or R.sup.3aR.sup.3b, wherein R.sup.3a and
R.sup.3b are each hydroxy.
[0110] Also provided herein are compounds of Formula II:
##STR00010##
wherein:
[0111] X is O, S, NH, NMe or CR.sup.xR.sup.y, wherein R.sup.x and
R.sup.y are each independently selected from H, alkyl and halo;
[0112] n is 0, 1, 2, 3, 4, 5 or 6;
[0113] m is 0, 1, 2, or 3;
[0114] R.sup.1 is selected from the group consisting of: alkyl,
alkenyl, alkynyl, aryl, halo, hydroxy, amine and carboxy;
[0115] R.sup.2 is selected from the group consisting of: H, alkyl
(e.g., lower alkyl), hydroxy, amine, and .dbd.O; or R.sup.2 is
R.sup.2aR.sup.2b, wherein R.sup.2a and R.sup.2b are each
independently selected from alkyl (e.g., lower alkyl) and
hydroxy;
[0116] R.sup.3 is selected from the group consisting of: H,
hydroxy, amine, and .dbd.O; or R.sup.3 is R.sup.3aR.sup.3b, wherein
R.sup.3a and R.sup.3b are each independently selected from alkyl
(e.g., lower alkyl) and hydroxy;
[0117] R.sup.4 is selected from the group consisting of: H, alkyl
(e.g., lower alkyl), and hydroxy; or R.sup.4 is R.sup.4aR.sup.4b
wherein R.sup.4a and R.sup.4b are each independently selected from
alkyl (e.g., lower alkyl), hydroxy, and halo, wherein said alkyl
may be unsubstituted or substituted 1, 2 or 3 times with hydroxy;
and
[0118] each R.sup.5 is independently selected from the group
consisting of: H, alkyl (e.g., lower alkyl), hydroxy, amine, and
.dbd.O; or R.sup.5a is R.sup.5aR.sup.5b wherein R.sup.5a and
R.sup.5b are each independently selected from alkyl (e.g., lower
alkyl) and hydroxy; or R.sup.2 and an adjacent R.sup.5 are taken
together to form an aryl or heteroaryl,
[0119] or a pharmaceutically acceptable salt or prodrug
thereof.
[0120] In some embodiments of Formula II, X is O, NH, NMe or
CR.sup.xR.sup.y.
[0121] In some embodiments of Formula II, n is 0 and/or R.sup.1 is
hydroxy.
[0122] In some embodiments of Formula II, R.sup.2 and/or R.sup.1 is
selected from the group consisting of: H and lower alkyl.
[0123] In some embodiments of Formula II, R.sup.3 is hydroxy.
[0124] In some embodiments of Formula II, R.sup.2 is selected from
the group consisting of: H, hydroxy, and lower alkyl.
[0125] Further provided herein are compounds of Formula II:
##STR00011##
wherein:
[0126] X is O, S, NH, NMe or CR.sup.xR.sup.y, wherein R.sup.x and
R.sup.y are each independently selected from H, alkyl and halo;
[0127] n is 0, 1, 2, 3, 4, 5 or 6;
[0128] R.sup.1 is selected from the group consisting of: alkyl,
alkenyl, alkynyl, aryl, halo, hydroxy, amine and carboxy;
[0129] R.sup.2 is selected from the group consisting of: H, alkyl
(e.g., lower alkyl), hydroxy, amine, and .dbd.O; or R.sup.2 is
R.sup.2aR.sup.2b, wherein R.sup.2a and R.sup.2b are each
independently selected from alkyl (e.g., lower alkyl) and
hydroxy;
[0130] R.sup.3 is selected from the group consisting of: H,
hydroxy, amine, and .dbd.O; or R.sup.3 is R.sup.3aR.sup.3b, wherein
R.sup.3a and R.sup.3b are each independently selected from alkyl
(e.g., lower alkyl) and hydroxy;
[0131] R.sup.4 is selected from the group consisting of: H, alkyl
(e.g., lower alkyl), and hydroxy; or R.sup.4 is R.sup.4aR.sup.4b
wherein R.sup.4a and R.sup.4b are each independently selected from
alkyl (e.g., lower alkyl), hydroxy, and halo, wherein said alkyl
may be unsubstituted or substituted 1, 2 or 3 times with hydroxy;
and
[0132] R.sup.5 is independently selected from the group consisting
of: H, alkyl (e.g., lower alkyl), hydroxy, amine, and .dbd.O; or
R.sup.5 is R.sup.5aR.sup.5b wherein R.sup.5a and R.sup.5b are each
independently selected from alkyl (e.g., lower alkyl) and
hydroxy,
[0133] or a pharmaceutically acceptable salt or prodrug
thereof.
[0134] In some embodiments of Formula III, X is NH.
[0135] In some embodiments of Formula III, n is 0 and/or R.sup.1 is
hydroxy.
[0136] In some embodiments of Formula I, R.sup.2 and/or R.sup.4 is
selected from the group consisting of: H and lower alkyl.
[0137] In some embodiments of Formula III, R.sup.3 is hydroxy.
2. Formulations.
[0138] The active compounds described herein may be formulated for
administration in a pharmaceutical carrier in accordance with known
techniques. See, e.g., Remington, The Science And Practice of
Pharmacy (9.sup.th Ed. 1995). In the manufacture of a
pharmaceutical formulation according to the invention, the active
compound (including the physiologically acceptable salts or
prodrugs thereof) is typically admixed with, inter alia, an
acceptable carrier. The carrier must, of course, be acceptable in
the sense of being compatible with any other ingredients in the
formulation and must not be deleterious to the patient. The carrier
may be a solid or a liquid, or both, and is preferably formulated
with the compound as a unit-dose formulation, for example, a
tablet, which may contain from 0.01 or 0.5% to 95% or 99% by weight
of the active agent. One or more active agents may be incorporated
in the formulations of the invention, which may be prepared by any
of the well-known techniques of pharmacy comprising admixing the
components, optionally including one or more accessory
ingredients.
[0139] The pharmaceutical compositions may also contain other
additives, such as pH-adjusting additives. In particular, useful
pH-adjusting agents include acids, such as hydrochloric acid, bases
and/or buffers, such as sodium lactate, sodium acetate, sodium
phosphate, sodium citrate, sodium borate, or sodium gluconate.
Further, the compositions may contain preservatives. Useful
preservatives include methylparaben, propylparaben, benzoic acid
and benzyl alcohol.
[0140] Formulations of the invention include those suitable for
oral, buccal (sub-lingual), parenteral (e.g., subcutaneous,
intramuscular, intradermal, or intravenous), topical (i.e., both
skin and mucosal surfaces, including airway surfaces) and
transdermal administration, although the most suitable route in any
given case will depend on the nature and severity of the condition
being treated and on the nature of the particular active compound
being used.
[0141] Formulations suitable for oral administration may be
presented in discrete units, such as capsules, cachets, lozenges,
or tablets, each containing a predetermined amount of the active
compound(s); as a powder or granules; as a solution or a suspension
in an aqueous or non-aqueous liquid; or as an oil-in-water or
water-in-oil emulsion. Such formulations may be prepared by any
suitable method of pharmacy which includes the step of bringing
into association the active compound and a suitable carrier (which
may contain one or more accessory ingredients as noted above). In
general, the formulations of the invention are prepared by
uniformly and intimately admixing the active compound with a liquid
or finely divided solid carrier, or both, and then, if necessary,
shaping the resulting mixture. For example, a tablet may be
prepared by compressing or molding a powder or granules containing
the active compound, optionally with one or more accessory
ingredients. Compressed tablets may be prepared by compressing, in
a suitable machine, the compound in a free-flowing form, such as a
powder or granules optionally mixed with a binder, lubricant, inert
diluent, and/or surface active/dispersing agent(s). Molded tablets
may be made by molding, in a suitable machine, the powdered
compound moistened with an inert liquid binder.
[0142] Formulations suitable for oral administration also include
food product formulations, such as a nutritional bar or an animal
feed (e.g., pet food such as dog or cat food). Food product
formulations may include one or more of carbohydrates such as
wheat, corn rice, barley or oats, dairy products such as milk, oils
such as canola oil or soybean oil, flavorants such as sugar or
syrup, coloring, chocolate, preservatives, etc. Pet food
formulations, in particular, may include meat, poultry, fish or
other animal-derived components such as eggs.
[0143] Formulations suitable for buccal (sub-lingual)
administration include lozenges comprising the active compound in a
flavored base, usually sucrose and acacia or tragacanth; and
pastilles comprising the compound in an inert base such as gelatin
and glycerin or sucrose and acacia.
[0144] Formulations of the present invention suitable for
parenteral administration comprise sterile aqueous and non-aqueous
injection solutions, which preparations are preferably isotonic
with the blood of the intended recipient. These preparations may
contain anti-oxidants, buffers, bacteriostats and solutes which
render the formulation isotonic with the blood of the intended
recipient. Aqueous and non-aqueous sterile suspensions may include
suspending agents and thickening agents. The formulations may be
presented in unit\dose or multi-dose containers, for example sealed
ampoules and vials, and may be stored in a freeze-dried
(lyophilized) condition requiring only the addition of the sterile
liquid carrier, for example, saline or water-for-injection
immediately prior to use. Extemporaneous injection solutions and
suspensions may be prepared from sterile powders, granules and
tablets of the kind previously described. For example, in one
aspect of the present invention, there is provided an injectable,
stable, sterile composition comprising an active compound(s) in a
unit dosage form in a sealed container. The active compound(s) may
be provided in the form of a lyophilizate which is capable of being
reconstituted with a suitable pharmaceutically acceptable carrier
to form a liquid composition suitable for injection thereof into a
subject.
[0145] When the active compound(s) is substantially
water-insoluble, a sufficient amount of emulsifying agent which is
physiologically acceptable may be employed in sufficient quantity
to emulsify the compound or salt in an aqueous carrier. One such
useful emulsifying agent is phosphatidyl choline.
[0146] Formulations suitable for topical application to the skin
preferably take the form of an ointment, cream, lotion, paste, gel,
spray, aerosol, or oil. Carriers which may be used include
petroleum jelly, lanoline, polyethylene glycols, alcohols,
transdermal enhancers, and combinations of two or more thereof.
[0147] Formulations suitable for transdermal administration may be
presented as discrete patches adapted to remain in intimate contact
with the epidermis of the recipient for a prolonged period of time.
Formulations suitable for transdermal administration may also be
delivered by iontophoresis (see, for example, Pharmaceutical
Research 3 (6):318 (1986)) and typically take the form of an
optionally buffered aqueous solution of the active compound.
Suitable formulations comprise citrate or bis\tris buffer (pH 6) or
ethanol/water and contain from 0.1 to 0.2M active ingredient.
[0148] The unit dosage form typically comprises from about 1 mg, 5
mg, 10 mg, 100 mg, 250 mg, 500 mg, 1 gram, 5 grams, 10 grams, or
any ranges therein, of the active compound(s), depending on the
subject being treated (e.g., human or non-human mammalian subject).
In some embodiments, the unit dosage form is in the range of 500 mg
to 10 grams, keeping in mind that a good portion of the active
compound(s) may not be absorbed upon administration (e.g., oral
administration).
[0149] The present invention is explained in greater detail in the
following non-limiting examples.
EXAMPLES
Example 1. Measurement of Hydroxyproline Metabolism
[0150] Patients with PH1, PH2, and PH3 and normal subjects were
placed on a 3-day controlled diet and infused in the fasted state
with .sup.15N--.sup.13C.sub.5-Hyp at a constant rate (750
nmol/kg/h) for 6 h. Urine and plasma samples were collected hourly
for analysis: total .sup.13C-labelled Hyp and glycine by GC/MS;
oxalate and glycolate by IC and IC/MS. The tracer has proven to
work effectively and safely. The tracer did not change the
pre-infusion and post-infusion total urinary oxalate excretion
(e.g., 13.+-.3 versus 9.+-.4 mg/g creat/h for normal; 60.+-.50
versus 40.+-.29 mg/g creat/h for PH1; similar values for PH2 and
PH3 samples).
[0151] A preliminary comparison of the enrichment of the tracer in
plasma Hyp, urine oxalate, and urine glycolate reveals intriguing
patterns and highlights the degree to which Hyp metabolism in PH1-3
patients is different. The preliminary data indicate that the
plasma levels of .sup.15N--.sup.13C.sub.5-Hyp in PH1 and PH3
patients is enriched over controls (.about.2-fold). This may
suggest that Hyp turnover is slower in these patients; however, the
range of values for the patients tested is quite wide, and overlaps
with the control values (hence, the need to know which PH1
mutations are present and the treatment regimen). There is also the
possibility that collagen breakdown by collagenases may be yielding
a spectrum of peptides that may be metabolized more quickly and
partition differently in plasma than free Hyp.
[0152] Notably, the proportion of the label in urine oxalate is
significantly increased in all PH patient groups (2- to 8-fold),
with PH3 being the highest. This observation supports that HOG is
being broken down in PH3 patients by another pathway to
glyoxylate/oxalate.
[0153] Altogether, these observations suggest that Hyp contributes
up to 25% of urinary oxalate.
[0154] An increase in the level of urine oxalate, on the order of
3-5 mg/day, can have up to a 2-fold increase in stone disease risk
(Zhang, BMC Bioinformatics 9, 40, 2008). Therefore, blocking HYPDH
activity has the potential to decrease the amount of glyoxylate and
oxalate produced endogenously by all three types of PH patients,
and to markedly reduce their risk for stone formation and disease.
Similarly, HYPDH inhibition may also benefit idiopathic stone
formers and other individuals with high urinary oxalate levels,
such as those that have undergone gastric bypass surgery. For the
latter, there is a significant increase in stone formation that may
benefit from prophylactic treatment post surgery. While the exact
origins of the oxalate in these patients has not been determined,
inhibition of HYPDH will decrease glyoxylate and oxalate levels,
which will ultimately reduce the glyoxylate and oxalate burden in
them.
Example 2. Development of Recombinant, Human HYPDH and Activity
Assay
[0155] Despite the identification of the Hyp pathway in rat and
bovine kidneys and livers over 50 years ago, very little is known
about human HYPDH (also known as PRODH2 and hydroxyproline oxidase,
HPOX, in the literature) (Miyata et al., Proc Natl Acad Sci USA
111, 14406-14411, 2014; Efron et al., New Engl J Med 272,
1299-1309; Pelkonen et al., New Engl J Med 283, 451-456, 1970). In
an effort to biochemically and structurally characterize human
HYPDH, we have evaluated numerous expression constructs (>15) in
Escherichia coli with N- and C-terminal truncations. These
constructs exhibit different levels of protein production,
solubility (i.e., inclusion body formation), FAD+ cofactor loading,
and enzymatic activity. Only the constructs containing the residues
147-515 and 156-515 were >96% loaded with FAD+ and active.
[0156] Recombinant HYPDH: (1) displays typical FAD spectra upon
oxidation and reduction, (2) exhibits kinetic parameters for the
turnover of Hyp consistent with homologs (Zhang, BMC Bioinformatics
9, 40, 2008; Moxley et al., Biochemistry 51, 511-520, 2012; Moxley
et al., Arch Biochem Biophys 516, 113-120, 2011; Srivastava et al.,
Proc Natl Acad Sci USA 107, 2878-2883, 2010), (3) is selective for
Hyp and not Pro, (4) readily uses a variety of CoQ10 analogs as an
electron acceptor during catalysis, and (5) binds Hyp with a KD
value of 125 .mu.M, using an anaerobic titration of the FAD+
spectrum. These data represent the first biochemical data available
for human HYPDH by any laboratory.
Example 3. Identification and Testing of Inhibitors of HYPDH
[0157] Tested compounds are shown in FIG. 2. Some compounds were
commercially available, and non-commercial compounds were
synthesized on a fee-for-service basis contract with Jasco
Pharmaceuticals (Woburn, Mass.). Compound 3 is not yet tested, and
compound 4 is not yet synthesized.
[0158] Each inhibitor was pre-incubated with HYPDH for 5 min, and
the reaction started by the addition of 600 mM Hyp. A range of
concentrations was tested in order to determine the IC50 value.
Table 1 lists the potency of the compounds.
TABLE-US-00001 TABLE 1 Cmpd IC.sub.50 (mM) 1 2.9 .+-. 0.1 5 2.1
.+-. 0.1 2 1.5 .+-. 0.1 3 ND.sup.a 4 ND.sup.b 9 0.63 .+-. 0.01 6
>10 7 0.60 .+-. 0.01 10 0.38 .+-. 0.01 11 0.37 .+-. 0.01 8 0.48
.+-. 0.01 13 0.32 .+-. 0.01 12 <1 15 0.37 .+-. 0.01 14 0.33 .+-.
0.01 16 0.29 .+-. 0.01 .sup.aNot soluble in buffer .sup.bSynthesis
planned
[0159] Inhibitors of HYPDH were identified as compounds in which
the nitrogen atom of the Hyp ring is changed to oxygen, carbon or
sulfur. This substitution prevents ring oxidation and cleavage by
HYPDH. The data indicate that the most potent compounds belong to
the reduced thiophene class, closely followed by the cyclopentane
analogs.
Example 4. Further Testing of Inhibitors of HYPDH
[0160] Additional compounds are obtained, and tested in the same
manner as in Example 3 above. These additional compounds may
include those in Scheme 1:
##STR00012##
Additional compounds may also include those in Scheme 2:
##STR00013##
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[0194] The foregoing is illustrative of the present invention, and
is not to be construed as limiting thereof. The invention is
defined by the following claims, with equivalents of the claims to
be included therein.
* * * * *