U.S. patent application number 14/349010 was filed with the patent office on 2014-10-16 for methods and compositions comprising a nitrite-reductase promoter for treatment of medical disorders and preservation of blood products.
The applicant listed for this patent is RadioRx, Inc.. Invention is credited to Chad Brouse, Pedro Cabrales, Marcel Fens, William Fitch, Susan Knox, Frans A. Kuypers, Sandra Larkin, Bryan T. Oronsky, Jan Scicinski.
Application Number | 20140308260 14/349010 |
Document ID | / |
Family ID | 48044412 |
Filed Date | 2014-10-16 |
United States Patent
Application |
20140308260 |
Kind Code |
A1 |
Oronsky; Bryan T. ; et
al. |
October 16, 2014 |
METHODS AND COMPOSITIONS COMPRISING A NITRITE-REDUCTASE PROMOTER
FOR TREATMENT OF MEDICAL DISORDERS AND PRESERVATION OF BLOOD
PRODUCTS
Abstract
The invention provides methods, compositions, and medical kits
comprising a nitrite-reductase promoter, such as an allosteric
modulator of hemoglobin, for use in treating medical disorders and
preservation of blood products. In one aspect, the invention
provides methods, compositions, and medical kits comprising an
inorganic nitrite salt and a nitrite-reductase promoter, such as an
allosteric modulator of hemoglobin, for use in treating medical
disorders, such as cancer, cardiovascular disorders, ischemic
conditions, hemolytic conditions, and bacterial infections.
Exemplary inorganic nitrite salts include sodium nitrite and
arginine nitrite. Exemplary allosteric modulators of hemoglobin
described herein include alkyl-substituted and acyl-substituted
di-nitroheterocycles.
Inventors: |
Oronsky; Bryan T.; (Los
Altos Hills, CA) ; Scicinski; Jan; (Saratoga, CA)
; Knox; Susan; (Stanford, CA) ; Fitch;
William; (Palo Alto, CA) ; Kuypers; Frans A.;
(El Cerrito, CA) ; Fens; Marcel; (San Francisco,
CA) ; Larkin; Sandra; (Berkeley, CA) ;
Cabrales; Pedro; (San Diego, CA) ; Brouse; Chad;
(San Diego, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
RadioRx, Inc. |
Mountain View |
CA |
US |
|
|
Family ID: |
48044412 |
Appl. No.: |
14/349010 |
Filed: |
October 5, 2012 |
PCT Filed: |
October 5, 2012 |
PCT NO: |
PCT/US12/58964 |
371 Date: |
April 1, 2014 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
61544375 |
Oct 7, 2011 |
|
|
|
Current U.S.
Class: |
424/93.71 ;
424/718; 424/93.7; 424/93.73; 435/2; 514/13.5; 530/385; 600/1 |
Current CPC
Class: |
A61K 38/1709 20130101;
A61K 31/255 20130101; A61K 31/215 20130101; A61K 31/445 20130101;
A61K 47/6445 20170801; A61K 35/18 20130101; A61K 31/22 20130101;
A61K 33/00 20130101; A61K 31/397 20130101; A61K 31/40 20130101;
A61K 31/12 20130101; A61K 31/198 20130101; A61K 35/14 20130101;
A61K 45/06 20130101; A61N 5/10 20130101; A01N 1/0226 20130101; Y02A
50/411 20180101; Y02A 50/30 20180101; A61K 31/16 20130101; A61K
35/16 20130101; A61K 33/00 20130101; A61K 2300/00 20130101; A61K
31/397 20130101; A61K 2300/00 20130101; A61K 31/40 20130101; A61K
2300/00 20130101; A61K 31/445 20130101; A61K 2300/00 20130101; A61K
31/12 20130101; A61K 2300/00 20130101; A61K 31/22 20130101; A61K
2300/00 20130101; A61K 31/215 20130101; A61K 2300/00 20130101; A61K
31/255 20130101; A61K 2300/00 20130101; A61K 31/16 20130101; A61K
2300/00 20130101; A61K 31/198 20130101; A61K 2300/00 20130101 |
Class at
Publication: |
424/93.71 ;
424/93.7; 424/93.73; 424/718; 514/13.5; 530/385; 435/2; 600/1 |
International
Class: |
A61K 31/397 20060101
A61K031/397; A61K 35/14 20060101 A61K035/14; A61K 35/18 20060101
A61K035/18; A61N 5/10 20060101 A61N005/10; A61K 38/17 20060101
A61K038/17; A61K 45/06 20060101 A61K045/06; A01N 1/02 20060101
A01N001/02; A61K 35/16 20060101 A61K035/16; A61K 33/00 20060101
A61K033/00 |
Claims
1. A method of treating or preventing a disorder selected from the
group consisting of cancer, a cardiovascular disorder, an ischemic
condition, a hemolytic condition, or a bacterial infection,
comprising administering to a patient in need thereof a
therapeutically effective amount of (i) an inorganic nitrite salt,
and (ii) an allosteric modulator of hemoglobin that promotes
nitrite reductase activity.
2. The method of claim 1, wherein the disorder is cancer.
3. The method of claim 2, wherein the cancer is a tumor.
4. The method of claim 2 or 3, further comprising exposing the
patient to a chemotherapeutic agent or radiation.
5. The method of claim 1, wherein the disorder is a cardiovascular
disorder.
6. The method of claim 5, wherein the cardiovascular disorder is
pulmonary hypertension, systemic hypertension, angina, Cardiac
Syndrome X, myocardial infarction, peripheral artery disease, or
Raynaud's disease.
7. The method of claim 1, wherein the disorder is a hemolytic
condition.
8. The method of claim 7, wherein the hemolytic condition is sickle
cell disease.
9. A method of increasing the amount of nitric oxide produced by
hemoglobin in a patient, comprising administering to a patient in
need thereof a therapeutically effective amount of (i) an inorganic
nitrite salt, and (ii) an allosteric modulator of hemoglobin that
promotes nitrite reductase activity.
10. The method of any one of claims 1-9, wherein the inorganic
nitrite salt is an alkali metal nitrite.
11. The method of any one of claims 1-9, wherein the inorganic
nitrite salt is sodium nitrite.
12. The method of any one of claims 1-9, wherein the inorganic
nitrite salt is represented by NO.sub.2--N(R').sub.4, wherein R'
represents independently for each occurrence hydrogen, optionally
substituted alkyl, optionally substituted heteroalkyl, optionally
substituted cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, optionally substituted aralkyl,
optionally substituted heteroaryl, or optionally substituted
heteroaralkyl.
13. The method of any one of claims 1-12, wherein the allosteric
modulator of hemoglobin binds to the beta-cysteine-93 residue of
hemoglobin.
14. The method of any one of claims 1-12, wherein the allosteric
modulator of hemoglobin is a compound of Formula I or II, wherein
Formula I is represented by: ##STR00173## or a pharmaceutically
acceptable salt or solvate thereof, wherein: A.sup.1 is --C(O)-- or
--(C(R.sup.3).sub.2).sub.xC(O)(C(R.sup.3).sub.2).sub.x--; A.sup.2
is N or --C(R.sup.4)--; R.sup.1 is halogen, --OS(O).sub.2R.sup.5,
or --OC(O)CF.sub.3; R.sup.2 is C.sub.1-C.sub.6alkyl; R.sup.3 and
R.sup.4 each represent independently for each occurrence hydrogen
or C.sub.1-C.sub.5alkyl; R.sup.5 is C.sub.1-C.sub.6alkyl,
C.sub.1-C.sub.6haloalkyl, aryl, or aralkyl; m and p are
independently 1, 2, or 3; and n and x each represent independently
for each occurrence 0, 1, 2, or 3; and Formula II is represented
by: ##STR00174## or a pharmaceutically acceptable salt or solvate
thereof: wherein: A.sup.1 is --C(O)-- or
--(C(R.sup.5).sub.2).sub.xC(O)(C(R.sup.5).sub.2).sub.x--; A.sup.2
is --N(R.sup.5)-- or --C(R.sup.2)(R.sup.3)--; R.sup.1 is halogen,
--OS(O).sub.2R.sup.6, or --OC(O)CF.sub.3; R.sup.2 and R.sup.3 each
represent independently for each occurrence hydrogen or
C.sub.1-C.sub.6alkyl; or R.sup.2 and R.sup.3 are taken together
with the carbon atom to which they are attached to form a 3-6
membered, saturated carbocyclic ring; R.sup.4 is hydrogen or
C.sub.1-C.sub.6alkyl; R.sup.5 represents independently for each
occurrence hydrogen or C.sub.1-C.sub.6alkyl; R.sup.6 is
C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6haloalkyl, aryl, or aralkyl; t
is an integer in the range from 1 to 12; and x represents
independently for each occurrence 0, 1, 2, or 3.
15. The method of any one of claims 1-12, wherein the allosteric
modulator of hemoglobin is a compound of Formula I-A: ##STR00175##
or a pharmaceutically acceptable salt or solvate thereof, wherein:
A is N or C(H); R.sup.1 is chloro, bromo,
--OS(O).sub.2--(C.sub.1-C.sub.6alkyl),
--OS(O).sub.2--(C.sub.1-C.sub.6haloalkyl),
--OS(O).sub.2-(para-methylphenyl), or --OC(O)CF.sub.3; R.sup.2
represents independently for each occurrence hydrogen or methyl;
and y represents independently for each occurrence 1 or 2.
16. The method of claim 15, wherein A is N.
17. The method of claim 15 or 16, wherein R.sup.1 is bromo.
18. The method of any one of claims 15-17, wherein y is 1.
19. The method of any one of claims 1-12, wherein the allosteric
modulator of hemoglobin is ##STR00176## or a pharmaceutically
acceptable salt or solvate thereof.
20. The method of any one of claims 1-12, wherein the allosteric
modulator of hemoglobin is selected from the group consisting of
S-nitroso-N-acetylcysteine, S-nitrosocysteinylglycine,
S-nitrosocysteine, S-nitrosohomocysteine, a metal nitrosyl complex,
an S-nitro compound, an S-nitroso compound, a thionitrite, a
diazeniumdiolate, 4-pyridylmethyl chloride, an alkoxyalkylchloride,
dimethoxymethane, N-(hydroxymethyl)acetamide, triphenylmethyl
chloride, acetyl chloride, 2-chloroacetic acid, acetic anhydride, a
haloacetamide, a haloacetate, benzyl chloride, benzoyl chloride,
di-tert-butyl dicarbonate, p-hydroxyphenacyl bromide,
p-acetoxybenzyl chloride, p-methoxybenzyl chloride,
tetrahydropyran, acetamidohydroxymethane, acetone,
bis-carboethoxyethene, tert-butoxycarbonyl chloride, alkyl
isocyanate, alkoxyalkyl isocyanate, a derivatized dextran, a
(polyethylene glycol)-maleimide, 2,4-dinitrophenyl fluoride, and
2,2,2-trichloroethoxycarbonyl.
21. The method of any one of claims 1-20, wherein the inorganic
nitrite salt is administered at a daily dosage of about 0.1
.mu.g/kg to about 10 mg/kg.
22. The method of any one of claims 1-21, wherein the inorganic
nitrite salt is administered orally.
23. The method of any one of claims 1-22, wherein the allosteric
modulator of hemoglobin is administered at a dosage sufficient to
cause a ten percent increase in the rate at which hemoglobin
converts nitrite to nitric oxide in vivo.
24. The method of any one of claims 1-23, wherein the allosteric
modulator is administered within about 1 hour after administration
of the inorganic nitrite salt.
25. A pharmaceutical composition comprising (i) an inorganic
nitrite salt, and (ii) an allosteric modulator of hemoglobin that
promotes nitrite reductase activity.
26. The pharmaceutical composition of claim 25, wherein the
inorganic nitrite salt is an alkali metal nitrite.
27. The pharmaceutical composition of claim 25, wherein the
inorganic nitrite salt is sodium nitrite.
28. The pharmaceutical composition of claim 25, wherein the
inorganic nitrite salt is represented by NO.sub.2--N(R').sub.4,
wherein R' represents independently for each occurrence hydrogen,
optionally substituted alkyl, optionally substituted heteroalkyl,
optionally substituted cycloalkyl, optionally substituted
heterocycloalkyl, optionally substituted aryl, optionally
substituted aralkyl, optionally substituted heteroaryl, or
optionally substituted heteroaralkyl.
29. The pharmaceutical composition of any one of claims 25-28,
wherein the allosteric modulator of hemoglobin binds to the
beta-cysteine-93 residue of hemoglobin.
30. The pharmaceutical composition of any one of claims 25-28,
wherein the allosteric modulator of hemoglobin is a compound of
Formula I or II, wherein Formula I is represented by: ##STR00177##
or a pharmaceutically acceptable salt or solvate thereof, wherein:
A.sup.1 is --C(O)-- or
--(C(R.sup.3).sub.2).sub.xC(O)(C(R.sup.3).sub.2).sub.x--; A.sup.2
is N or --C(R.sup.4)--; R.sup.1 is halogen, --OS(O).sub.2R.sup.5,
or --OC(O)CF.sub.3; R.sup.2 is C.sub.1-C.sub.6alkyl; R.sup.3 and
R.sup.4 each represent independently for each occurrence hydrogen
or C.sub.1-C.sub.5alkyl; R.sup.5 is C.sub.1-C.sub.6alkyl,
C.sub.1-C.sub.6haloalkyl, aryl, or aralkyl; m and p are
independently 1, 2, or 3; and n and x each represent independently
for each occurrence 0, 1, 2, or 3; and Formula II is represented
by: ##STR00178## or a pharmaceutically acceptable salt or solvate
thereof: wherein: A.sup.1 is --C(O)-- or
--(C(R.sup.5).sub.2).sub.xC(O)(C(R.sup.5).sub.2).sub.x--; A.sup.2
is --N(R.sup.5)-- or --C(R.sup.2)(R.sup.3)--; R.sup.1 is halogen,
--OS(O).sub.2R.sup.6, or --OC(O)CF.sub.3; R.sup.2 and R.sup.3 each
represent independently for each occurrence hydrogen or
C.sub.1-C.sub.6alkyl; or R.sup.2 and R.sup.3 are taken together
with the carbon atom to which they are attached to form a 3-6
membered, saturated carbocyclic ring; R.sup.4 is hydrogen or
C.sub.1-C.sub.6alkyl; R.sup.5 represents independently for each
occurrence hydrogen or C.sub.1-C.sub.6alkyl; R.sup.6 is
C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6haloalkyl, aryl, or aralkyl; t
is an integer in the range from 1 to 12; and x represents
independently for each occurrence 0, 1, 2, or 3.
31. The pharmaceutical composition of any one of claims 25-28,
wherein the allosteric modulator of hemoglobin is a compound of
Formula I-A: ##STR00179## or a pharmaceutically acceptable salt or
solvate thereof, wherein: A is N or C(H); R.sup.1 is chloro, bromo,
--OS(O).sub.2--(C.sub.1-C.sub.6alkyl),
--OS(O).sub.2--(C.sub.1-C.sub.6haloalkyl),
--OS(O).sub.2-(para-methylphenyl), or --OC(O)CF.sub.3; R.sup.2
represents independently for each occurrence hydrogen or methyl;
and y represents independently for each occurrence 1 or 2.
32. The pharmaceutical composition of claim 31, wherein A is N.
33. The pharmaceutical composition of claim 31 or 32, wherein
R.sup.1 is bromo.
34. The pharmaceutical composition of any one of claims 31-33,
wherein y is 1.
35. The pharmaceutical composition of any one of claims 25-28,
wherein the allosteric modulator of hemoglobin is ##STR00180## or a
pharmaceutically acceptable salt or solvate thereof.
36. The pharmaceutical composition of any one of claims 25-28,
wherein the allosteric modulator of hemoglobin is
S-nitroso-N-acetylcysteine, S-nitrosocysteinylglycine,
S-nitrosocysteine, S-nitrosohomocysteine, a metal nitrosyl complex,
an S-nitro compound, an S-nitroso compound, a thionitrite, a
diazeniumdiolate, 4-pyridylmethyl chloride, an alkoxyalkylchloride,
dimethoxymethane, N-(hydroxymethyl)acetamide, triphenylmethyl
chloride, acetyl chloride, 2-chloroacetic acid, acetic anhydride, a
haloacetamide, a haloacetate, benzyl chloride, benzoyl chloride,
di-tert-butyl dicarbonate, p-hydroxyphenacyl bromide,
p-acetoxybenzyl chloride, p-methoxybenzyl chloride,
tetrahydropyran, acetamidohydroxymethane, acetone,
bis-carboethoxyethene, tert-butoxycarbonyl chloride, alkyl
isocyanate, alkoxyalkyl isocyanate, a derivatized dextran, a
(polyethylene glycol)-maleimide, 2,4-dinitrophenyl fluoride, and
2,2,2-trichloroethoxycarbonyl.
37. A kit for treating a medical disorder, comprising (i) an
inorganic nitrite salt, (ii) an allosteric modulator of hemoglobin,
and (iii) instructions for using the kit to treat a medical
disorder.
38. A method of treating a patient suffering from reduced blood
volume, comprising administering to a patient in need thereof a
blood product by injection and a therapeutic agent selected from
the group consisting of an organonitro compound of Formula I,
organonitro compound of Formula II, hemoglobin conjugate of Formula
III, hemoglobin conjugate of Formula IV, and an erythrocyte cell
that has been exposed to an organonitro compound of Formula I or
II; wherein Formula I is represented by: ##STR00181## or a
pharmaceutically acceptable salt or solvate thereof, wherein:
A.sup.1 is --C(O)-- or
--(C(R.sup.3).sub.2).sub.xC(O)(C(R.sup.3).sub.2).sub.x--; A.sup.2
is N or --C(R.sup.4)--; R.sup.1 is halogen, --OS(O).sub.2R.sup.5,
or --OC(O)CF.sub.3; R.sup.2 is C.sub.1-C.sub.6alkyl; R.sup.3 and
R.sup.4 each represent independently for each occurrence hydrogen
or C.sub.1-C.sub.5alkyl; R.sup.5 is C.sub.1-C.sub.6alkyl,
C.sub.1-C.sub.6haloalkyl, aryl, or aralkyl; m and p are
independently 1, 2, or 3; and n and x each represent independently
for each occurrence 0, 1, 2, or 3; Formula II is represented by:
##STR00182## or a pharmaceutically acceptable salt or solvate
thereof: wherein: A.sup.1 is --C(O)-- or
--(C(R.sup.5).sub.2).sub.xC(O)(C(R.sup.5).sub.2).sub.x--; A.sup.2
is --N(R.sup.5)-- or --C(R.sup.2)(R.sup.3)--; R.sup.1 is halogen,
--OS(O).sub.2R.sup.6, or --OC(O)CF.sub.3; R.sup.2 and R.sup.3 each
represent independently for each occurrence hydrogen or
C.sub.1-C.sub.6alkyl; or R.sup.2 and R.sup.3 are taken together
with the carbon atom to which they are attached to form a 3-6
membered, saturated carbocyclic ring; R.sup.4 is hydrogen or
C.sub.1-C.sub.6alkyl; R.sup.5 represents independently for each
occurrence hydrogen or C.sub.1-C.sub.6alkyl; R.sup.6 is
C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6haloalkyl, aryl, or aralkyl; t
is an integer in the range from 1 to 12; and x represents
independently for each occurrence 0, 1, 2, or 3; Formula III is
represented by: ##STR00183## or a pharmaceutically acceptable salt
or solvate thereof, wherein: A.sup.1 is --C(O)-- or
--C(O)(C(R.sup.3).sub.2).sub.x--; A.sup.2 is N or --C(R.sup.4)--;
R.sup.2 is C.sub.1-C.sub.6alkyl; R.sup.3 and R.sup.4 each represent
independently for each occurrence hydrogen or C.sub.1-C.sub.5alkyl;
m and p are independently 1, 2, or 3; n is 0, 1, 2, or 3; x is 1,
2, or 3; and z is an integer from 1 to 10; and Formula IV is
represented by: ##STR00184## or a pharmaceutically acceptable salt
or solvate thereof, wherein: A.sup.1 is --C(O)-- or
--C(O)(C(R.sup.5).sub.2).sub.x--; A.sup.2 is --N(R.sup.5)-- or
--C(R.sup.2)(R.sup.3)--; R.sup.2 and R.sup.3 each represent
independently for each occurrence hydrogen or C.sub.1-C.sub.6alkyl;
or R.sup.2 and R.sup.3 are taken together with the carbon atom to
which they are attached to form a 3-6 membered, saturated
carbocyclic ring; R.sup.4 is hydrogen or C.sub.1-C.sub.6alkyl;
R.sup.5 represents independently for each occurrence hydrogen or
C.sub.1-C.sub.6alkyl; t is an integer in the range from 1 to 12; x
is 1, 2, or 3; and z is an integer from 1 to 10.
39. The method of claim 38, wherein the patient suffering from
reduced blood volume is suffering from hemorrhagic shock.
40. A method of performing a blood transfusion to a patient,
comprising administering to a patient in need thereof a blood
product by injection and a therapeutic agent selected from the
group consisting of an organonitro compound of Formula I,
organonitro compound of Formula II, hemoglobin conjugate of Formula
III, hemoglobin conjugate of Formula IV, and an erythrocyte cell
that has been exposed to an organonitro compound of Formula I or
II, wherein Formula I is represented by: ##STR00185## or a
pharmaceutically acceptable salt or solvate thereof, wherein:
A.sup.1 is --C(O)-- or
--(C(R.sup.3).sub.2).sub.xC(O)(C(R.sup.3).sub.2).sub.x--; A.sup.2
is N or --C(R.sup.4)--; R.sup.1 is halogen, --OS(O).sub.2R.sup.5,
or --OC(O)CF.sub.3; R.sup.2 is C.sub.1-C.sub.6alkyl; R.sup.3 and
R.sup.4 each represent independently for each occurrence hydrogen
or C.sub.1-C.sub.5alkyl; R.sup.5 is C.sub.1-C.sub.6alkyl,
C.sub.1-C.sub.6haloalkyl, aryl, or aralkyl; m and p are
independently 1, 2, or 3; and n and x each represent independently
for each occurrence 0, 1, 2, or 3; Formula II is represented by:
##STR00186## or a pharmaceutically acceptable salt or solvate
thereof: wherein: A.sup.1 is --C(O)-- or
--(C(R.sup.5).sub.2).sub.xC(O)(C(R.sup.5).sub.2).sub.x--; A.sup.2
is --N(R.sup.5)-- or --C(R.sup.2)(R.sup.3)--; R.sup.1 is halogen,
--OS(O).sub.2R.sup.6, or --OC(O)CF.sub.3; R.sup.2 and R.sup.3 each
represent independently for each occurrence hydrogen or
C.sub.1-C.sub.6alkyl; or R.sup.2 and R.sup.3 are taken together
with the carbon atom to which they are attached to form a 3-6
membered, saturated carbocyclic ring; R.sup.4 is hydrogen or
C.sub.1-C.sub.6alkyl; R.sup.5 represents independently for each
occurrence hydrogen or C.sub.1-C.sub.6alkyl; R.sup.6 is
C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6haloalkyl, aryl, or aralkyl; t
is an integer in the range from 1 to 12; and x represents
independently for each occurrence 0, 1, 2, or 3; Formula III is
represented by: ##STR00187## or a pharmaceutically acceptable salt
or solvate thereof, wherein: A.sup.1 is --C(O)-- or
--C(O)(C(R.sup.3).sub.2).sub.x--; A.sup.2 is N or --C(R.sup.4)--;
R.sup.2 is C.sub.1-C.sub.6alkyl; R.sup.3 and R.sup.4 each represent
independently for each occurrence hydrogen or C.sub.1-C.sub.5alkyl;
m and p are independently 1, 2, or 3; n is 0, 1, 2, or 3; x is 1,
2, or 3; and z is an integer from 1 to 10; and Formula IV is
represented by: ##STR00188## or a pharmaceutically acceptable salt
or solvate thereof, wherein: A.sup.1 is --C(O)-- or
--C(O)(C(R.sup.5).sub.2).sub.x--; A.sup.2 is --N(R.sup.5)-- or
--C(R.sup.2)(R.sup.3)--; R.sup.2 and R.sup.3 each represent
independently for each occurrence hydrogen or C.sub.1-C.sub.6alkyl;
or R.sup.2 and R.sup.3 are taken together with the carbon atom to
which they are attached to form a 3-6 membered, saturated
carbocyclic ring; R.sup.4 is hydrogen or C.sub.1-C.sub.6alkyl;
R.sup.5 represents independently for each occurrence hydrogen or
C.sub.1-C.sub.6alkyl; t is an integer in the range from 1 to 12; x
is 1, 2, or 3; and z is an integer from 1 to 10.
41. The method of any one of claims 38-40, wherein the blood
product comprises erythrocyte cells.
42. The method of any one of claims 38-41, wherein the blood
product comprises blood plasma.
43. The method of any one of claims 38-42, wherein the blood
product and organonitro compound are administered to the patient
concurrently.
44. The method of any one of claims 38-42, wherein the blood
product is administered to the patient separately from the
therapeutic agent.
45. The method of any one of claims 38-42, wherein the patient
receives, by intravenous injection, a single composition comprising
blood product and the therapeutic agent.
46. The method of any one of claims 38-45, further comprising
administering an alkali metal nitrite to the patient.
47. The method of any one of claims 38-45, further comprising
administering sodium nitrite to the patient.
48. A method of treating a patient suffering from anemia,
comprising administering to a patient in need thereof a therapeutic
agent selected from the group consisting of an organonitro compound
of Formula I, organonitro compound of Formula II, hemoglobin
conjugate of Formula III, hemoglobin conjugate of Formula IV, and
an erythrocyte cell that has been exposed to an organonitro
compound of Formula I or II; wherein Formula I is represented by:
##STR00189## or a pharmaceutically acceptable salt or solvate
thereof, wherein: A.sup.1 is --C(O)-- or
--(C(R.sup.3).sub.2).sub.xC(O)(C(R.sup.3).sub.2).sub.x--; A.sup.2
is N or --C(R.sup.4)--; R.sup.1 is halogen, --OS(O).sub.2R.sup.5,
or --OC(O)CF.sub.3; R.sup.2 is C.sub.1-C.sub.6alkyl; R.sup.3 and
R.sup.4 each represent independently for each occurrence hydrogen
or C.sub.1-C.sub.5alkyl; R.sup.5 is C.sub.1-C.sub.6alkyl,
C.sub.1-C.sub.6haloalkyl, aryl, or aralkyl; m and p are
independently 1, 2, or 3; and n and x each represent independently
for each occurrence 0, 1, 2, or 3; Formula II is represented by:
##STR00190## or a pharmaceutically acceptable salt or solvate
thereof: wherein: A.sup.1 is --C(O)-- or
--(C(R.sup.5).sub.2).sub.xC(O)(C(R.sup.5).sub.2).sub.x--; A.sup.2
is --N(R.sup.5)-- or --C(R.sup.2)(R.sup.3)--; R.sup.1 is halogen,
--OS(O).sub.2R.sup.6, or --OC(O)CF.sub.3; R.sup.2 and R.sup.3 each
represent independently for each occurrence hydrogen or
C.sub.1-C.sub.6alkyl; or R.sup.2 and R.sup.3 are taken together
with the carbon atom to which they are attached to form a 3-6
membered, saturated carbocyclic ring; R.sup.4 is hydrogen or
C.sub.1-C.sub.6alkyl; R.sup.5 represents independently for each
occurrence hydrogen or C.sub.1-C.sub.6alkyl; R.sup.6 is
C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6haloalkyl, aryl, or aralkyl; t
is an integer in the range from 1 to 12; and x represents
independently for each occurrence 0, 1, 2, or 3; Formula III is
represented by: ##STR00191## or a pharmaceutically acceptable salt
or solvate thereof, wherein: A.sup.1 is --C(O)-- or
--C(O)(C(R.sup.3).sub.2).sub.x--; A.sup.2 is N or --C(R.sup.4)--;
R.sup.2 is C.sub.1-C.sub.6alkyl; R.sup.3 and R.sup.4 each represent
independently for each occurrence hydrogen or C.sub.1-C.sub.5alkyl;
m and p are independently 1, 2, or 3; n is 0, 1, 2, or 3; x is 1,
2, or 3; and z is an integer from 1 to 10; and Formula IV is
represented by: ##STR00192## or a pharmaceutically acceptable salt
or solvate thereof, wherein: A.sup.1 is --C(O)-- or
--C(O)(C(R.sup.5).sub.2).sub.x--; A.sup.2 is --N(R.sup.5)-- or
--C(R.sup.2)(R.sup.3)--; R.sup.2 and R.sup.3 each represent
independently for each occurrence hydrogen or C.sub.1-C.sub.6alkyl;
or R.sup.2 and R.sup.3 are taken together with the carbon atom to
which they are attached to form a 3-6 membered, saturated
carbocyclic ring; R.sup.4 is hydrogen or C.sub.1-C.sub.6alkyl;
R.sup.5 represents independently for each occurrence hydrogen or
C.sub.1-C.sub.6alkyl; t is an integer in the range from 1 to 12; x
is 1, 2, or 3; and z is an integer from 1 to 10.
49. The method of claim 48, further comprising administering a
blood product to the patient by injection.
50. The method of claim 49, wherein the blood product comprises
erythrocyte cells.
51. The method of claim 49 or 50, wherein the blood product
comprises blood plasma.
52. The method of any one of claims 49-51, wherein the blood
product and organonitro compound are administered to the patient
concurrently.
53. The method of any one of claims 49-51, wherein the blood
product is administered to the patient separately from the
therapeutic agent.
54. The method of any one of claims 49-51, wherein the patient
receives, by intravenous injection, a single composition comprising
blood product and the therapeutic agent.
55. The method of any one of claims 48-54, further comprising
administering an alkali metal nitrite to the patient.
56. The method of any one of claims 48-54, further comprising
administering sodium nitrite to the patient.
57. The method of any one of claims 38-56, wherein the therapeutic
agent is an organonitro compound of Formula I.
58. The method of any one of claims 38-56, wherein the therapeutic
agent is an erythrocyte cell that has been exposed to an
organonitro compound of Formula I, and said therapeutic agent is
administered by injection.
59. The method of claim 57 or 58, wherein A.sup.1 is --C(O)--, and
A.sup.2 is N.
60. The method of any one of claims 38-59, wherein R.sup.1 is
bromo.
61. The method of any one of claims 38-60, wherein n is 0, and m is
2.
62. The method of any one of claims 38-58, wherein the therapeutic
agent is ##STR00193## or a pharmaceutically acceptable salt
thereof.
63. The method of any one of claims 38-56, wherein the therapeutic
agent is a hemoglobin conjugate of Formula III, and said
therapeutic agent is administered by injection.
64. The method of claim 63, wherein A.sup.1 is --C(O)-- and A.sup.2
is N.
65. The method of claim 63 or 64, wherein n is 0, and m is 2.
66. The method of any one of claims 38-56, wherein the therapeutic
agent is ##STR00194## or a pharmaceutically acceptable salt
thereof, wherein z is an integer from 1 to 10.
67. A method of preserving an isolated blood product, comprising
exposing the isolated blood product to an agent selected from the
group consisting of an organonitro compound of Formula I,
organonitro compound of Formula II, hemoglobin conjugate of Formula
III, hemoglobin conjugate of Formula IV, and an erythrocyte cell
that has been exposed to an organonitro compound of Formula I or
II, wherein Formula I is represented by: ##STR00195## or a
pharmaceutically acceptable salt or solvate thereof, wherein:
A.sup.1 is --C(O)-- or
--(C(R.sup.3).sub.2).sub.xC(O)(C(R.sup.3).sub.2).sub.x--; A.sup.2
is N or --C(R.sup.4)--; R.sup.1 is halogen, --OS(O).sub.2R.sup.5,
or --OC(O)CF.sub.3; R.sup.2 is C.sub.1-C.sub.6alkyl; R.sup.3 and
R.sup.4 each represent independently for each occurrence hydrogen
or C.sub.1-C.sub.5alkyl; R.sup.5 is C.sub.1-C.sub.6alkyl,
C.sub.1-C.sub.6haloalkyl, aryl, or aralkyl; m and p are
independently 1, 2, or 3; and n and x each represent independently
for each occurrence 0, 1, 2, or 3; Formula II is represented by:
##STR00196## or a pharmaceutically acceptable salt or solvate
thereof: wherein: A.sup.1 is --C(O)-- or
--(C(R.sup.5).sub.2).sub.xC(O)(C(R.sup.5).sub.2).sub.x--; A.sup.2
is --N(R.sup.5)-- or --C(R.sup.2)(R.sup.3)--; R.sup.1 is halogen,
--OS(O).sub.2R.sup.6, or --OC(O)CF.sub.3; R.sup.2 and R.sup.3 each
represent independently for each occurrence hydrogen or
C.sub.1-C.sub.6alkyl; or R.sup.2 and R.sup.3 are taken together
with the carbon atom to which they are attached to form a 3-6
membered, saturated carbocyclic ring; R.sup.4 is hydrogen or
C.sub.1-C.sub.6alkyl; R.sup.5 represents independently for each
occurrence hydrogen or C.sub.1-C.sub.6alkyl; R.sup.6 is
C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6haloalkyl, aryl, or aralkyl; t
is an integer in the range from 1 to 12; and x represents
independently for each occurrence 0, 1, 2, or 3; Formula III is
represented by: ##STR00197## or a pharmaceutically acceptable salt
or solvate thereof, wherein: A.sup.1 is --C(O)-- or
--C(O)(C(R.sup.3).sub.2).sub.x--; A.sup.2 is N or --C(R.sup.4)--;
R.sup.2 is C.sub.1-C.sub.6alkyl; R.sup.3 and R.sup.4 each represent
independently for each occurrence hydrogen or C.sub.1-C.sub.5alkyl;
m and p are independently 1, 2, or 3; n is 0, 1, 2, or 3; x is 1,
2, or 3; and z is an integer from 1 to 10; and Formula IV is
represented by: ##STR00198## or a pharmaceutically acceptable salt
or solvate thereof, wherein: A.sup.1 is --C(O)-- or
--C(O)(C(R.sup.5).sub.2).sub.x--; A.sup.2 is --N(R.sup.5)-- or
--C(R.sup.2)(R.sup.3)--; R.sup.2 and R.sup.3 each represent
independently for each occurrence hydrogen or C.sub.1-C.sub.6alkyl;
or R.sup.2 and R.sup.3 are taken together with the carbon atom to
which they are attached to form a 3-6 membered, saturated
carbocyclic ring; R.sup.4 is hydrogen or C.sub.1-C.sub.6alkyl;
R.sup.5 represents independently for each occurrence hydrogen or
C.sub.1-C.sub.6alkyl; t is an integer in the range from 1 to 12; x
is 1, 2, or 3; and z is an integer from 1 to 10.
68. The method of claim 67, wherein the isolated blood product is
whole blood.
69. The method of claim 67, wherein the isolated blood product
comprises erythrocyte cells.
70. The method of claim 67, wherein the isolated blood product is
erythrocyte cells.
71. The method of any one of claims 67-70, further comprising
exposing the isolated blood product to an alkali metal nitrite.
72. The method of any one of claims 67-70, further comprising
exposing the isolated blood product to sodium nitrite.
73. The method of any one of claims 67-72, wherein the agent is an
organonitro compound of Formula I.
74. The method of any one of claims 67-72, wherein the agent is an
erythrocyte cell that has been exposed to an organonitro compound
of Formula I.
75. The method of claim 73 or 74, wherein A.sup.1 is --C(O)--, and
A.sup.2 is N.
76. The method of any one of claims 67-75, wherein R.sup.1 is
bromo.
77. The method of any one of claims 67-76, wherein n is 0, and m is
2.
78. The method of any one of claims 67-72, wherein the agent is
##STR00199## or a pharmaceutically acceptable salt thereof.
79. The method of any one of claims 67-72, wherein the agent is a
hemoglobin conjugate of Formula III.
80. The method of claim 79, wherein A.sup.1 is --C(O)--, and
A.sup.2 is N.
81. The method of claim 79 or 80, wherein n is 0, and m is 2.
82. The method of any one of claims 67-72, wherein the agent is
##STR00200## or a pharmaceutically acceptable salt thereof, wherein
z is an integer from 1 to 10.
83. An isolated blood product composition, comprising (i) a blood
product, and (ii) an agent selected from the group consisting of an
organonitro compound of Formula I, organonitro compound of Formula
II, hemoglobin conjugate of Formula III, hemoglobin conjugate of
Formula IV, and an erythrocyte cell that has been exposed to an
organonitro compound of Formula I or II; wherein Formula I is
represented by: ##STR00201## or a pharmaceutically acceptable salt
or solvate thereof, wherein: A.sup.1 is --C(O)-- or
--(C(R.sup.3).sub.2).sub.xC(O)(C(R.sup.3).sub.2).sub.x--; A.sup.2
is N or --C(R.sup.4)--; R.sup.1 is halogen, --OS(O).sub.2R.sup.5,
or --OC(O)CF.sub.3; R.sup.2 is C.sub.1-C.sub.6alkyl; R.sup.3 and
R.sup.4 each represent independently for each occurrence hydrogen
or C.sub.1-C.sub.5alkyl; R.sup.5 is C.sub.1-C.sub.6alkyl,
C.sub.1-C.sub.6haloalkyl, aryl, or aralkyl; m and p are
independently 1, 2, or 3; and n and x each represent independently
for each occurrence 0, 1, 2, or 3; Formula II is represented by:
##STR00202## or a pharmaceutically acceptable salt or solvate
thereof: wherein: A.sup.1 is --C(O)-- or
--(C(R.sup.5).sub.2).sub.xC(O)(C(R.sup.5).sub.2).sub.x--; A.sup.2
is --N(R.sup.5)-- or --C(R.sup.2)(R.sup.3)--; R.sup.1 is halogen,
--OS(O).sub.2R.sup.6, or --OC(O)CF.sub.3; R.sup.2 and R.sup.3 each
represent independently for each occurrence hydrogen or
C.sub.1-C.sub.6alkyl; or R.sup.2 and R.sup.3 are taken together
with the carbon atom to which they are attached to form a 3-6
membered, saturated carbocyclic ring; R.sup.4 is hydrogen or
C.sub.1-C.sub.6alkyl; R.sup.5 represents independently for each
occurrence hydrogen or C.sub.1-C.sub.6alkyl; R.sup.6 is
C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6haloalkyl, aryl, or aralkyl; t
is an integer in the range from 1 to 12; and x represents
independently for each occurrence 0, 1, 2, or 3; Formula III is
represented by: ##STR00203## or a pharmaceutically acceptable salt
or solvate thereof, wherein: A.sup.1 is --C(O)-- or
--C(O)(C(R.sup.3).sub.2).sub.x--; A.sup.2 is N or --C(R.sup.4)--;
R.sup.2 is C.sub.1-C.sub.6alkyl; R.sup.3 and R.sup.4 each represent
independently for each occurrence hydrogen or C.sub.1-C.sub.5alkyl;
m and p are independently 1, 2, or 3; n is 0, 1, 2, or 3; x is 1,
2, or 3; and z is an integer from 1 to 10; and Formula IV is
represented by: ##STR00204## or a pharmaceutically acceptable salt
or solvate thereof, wherein: A.sup.1 is --C(O)-- or
--C(O)(C(R.sup.5).sub.2).sub.x--; A.sup.2 is --N(R.sup.5)-- or
--C(R.sup.2)(R.sup.3)--; R.sup.2 and R.sup.3 each represent
independently for each occurrence hydrogen or C.sub.1-C.sub.6alkyl;
or R.sup.2 and R.sup.3 are taken together with the carbon atom to
which they are attached to form a 3-6 membered, saturated
carbocyclic ring; R.sup.4 is hydrogen or C.sub.1-C.sub.6alkyl;
R.sup.5 represents independently for each occurrence hydrogen or
C.sub.1-C.sub.6alkyl; t is an integer in the range from 1 to 12; x
is 1, 2, or 3; and z is an integer from 1 to 10.
84. The composition of claim 83, wherein the blood product is whole
blood.
85. The composition of claim 83, wherein the blood product
comprises erythrocyte cells.
86. The composition of claim 83, wherein the blood product
comprises erythrocyte cells and blood plasma.
87. The composition of claim 83, wherein the blood product is
erythrocyte cells.
88. The composition of any one of claims 83-87, further comprising
an alkali metal nitrite.
89. The composition of any one of claims 83-87, further comprising
sodium nitrite.
90. The composition of any one of claims 83-89, wherein the agent
is an organonitro compound of Formula I.
91. The composition of any one of claims 83-89, wherein the agent
is an erythrocyte cell that has been exposed to an organonitro
compound of Formula I.
92. The composition of claim 90 or 91, wherein A.sup.1 is --C(O)--,
and A.sup.2 is N.
93. The composition of any one of claims 83-92, wherein R.sup.1 is
bromo.
94. The composition of any one of claims 83-93, wherein n is 0, and
m is 2.
95. The composition of any one of claims 83-89, wherein the agent
is ##STR00205## a pharmaceutically acceptable salt thereof.
96. The composition of any one of claims 83-89, wherein the agent
is a hemoglobin conjugate of Formula III.
97. The composition of claim 96, wherein A.sup.1 is --C(O)-- and
A.sup.2 is N.
98. The composition of claim 96 or 97, wherein n is 0, and m is
2.
99. The composition of any one of claims 83-89, wherein the agent
is ##STR00206## or a pharmaceutically acceptable salt thereof,
wherein z is an integer from 1 to 10.
100. An isolated hemoglobin conjugate represented by Formula III or
IV: ##STR00207## or a pharmaceutically acceptable salt or solvate
thereof, wherein: A.sup.1 is --C(O)-- or
--C(O)(C(R.sup.3).sub.2).sub.x--; A.sup.2 is N or --C(R.sup.4)--;
R.sup.2 is C.sub.1-C.sub.6alkyl; R.sup.3 and R.sup.4 each represent
independently for each occurrence hydrogen or C.sub.1-C.sub.5alkyl;
m and p are independently 1, 2, or 3; n is 0, 1, 2, or 3; x is 1,
2, or 3; and z is an integer from 1 to 10; and Formula IV is
represented by: ##STR00208## or a pharmaceutically acceptable salt
or solvate thereof, wherein: A.sup.1 is --C(O)-- or
--C(O)(C(R.sup.5).sub.2).sub.x--; A.sup.2 is --N(R.sup.5)-- or
--C(R.sup.2)(R.sup.3)--; R.sup.2 and R.sup.3 each represent
independently for each occurrence hydrogen or C.sub.1-C.sub.6alkyl;
or R.sup.2 and R.sup.3 are taken together with the carbon atom to
which they are attached to form a 3-6 membered, saturated
carbocyclic ring; R.sup.4 is hydrogen or C.sub.1-C.sub.6alkyl;
R.sup.5 represents independently for each occurrence hydrogen or
C.sub.1-C.sub.6alkyl; t is an integer in the range from 1 to 12; x
is 1, 2, or 3; and z is an integer from 1 to 10.
101. The isolated hemoglobin conjugate of claim 100, wherein the
agent is a hemoglobin conjugate of Formula III.
102. The isolated hemoglobin conjugate of claim 100 or 101, wherein
A.sup.1 is --C(O)--, and A.sup.2 is N.
103. The isolated hemoglobin conjugate of any one of claims
100-102, wherein n is 0, and m is 2.
104. The isolated hemoglobin conjugate of claim 100, wherein the
agent is ##STR00209## or a pharmaceutically acceptable salt
thereof, wherein z is an integer from 1 to 10.
105. The isolated hemoglobin conjugate of claim 104, wherein the
bond depicted to the hemoglobin is a thioether bond to the sulfur
atom of the beta-cysteine-93 residue of said hemoglobin.
106. A pharmaceutical composition, comprising a pharmaceutically
acceptable carrier and an isolated hemoglobin conjugate of claim
100.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application claims the benefit of and priority to U.S.
Provisional Patent Application Ser. No. 61/544,375, filed Oct. 7,
2011, the contents of which are hereby incorporated by
reference.
FIELD OF THE INVENTION
[0002] The invention provides methods, compositions, and medical
kits comprising a nitrite-reductase promoter, such as an allosteric
modulator of hemoglobin, for use in treating medical disorders and
preservation of blood products.
BACKGROUND
[0003] Cancer, cardiovascular disorders, ischemic conditions, and
bacterial infections remain a significant health problem for people
in many developed countries. The need for cancer treatments, for
example, has prompted the United States National Cancer Institute
to coordinate large-scale research efforts, impacting over six
hundred universities, hospitals, and cancer centers located in the
United States and over twenty foreign countries. Past research
efforts have lead to significant advances in the detection,
evaluation, and treatment of cancer, cardiovascular disorders,
ischemic conditions, and bacterial infections. However, despite
these developments, these medical conditions remain significant
health problems for many patients.
[0004] According to current statistics, cancer is a leading cause
of death worldwide. Approximately one million people are diagnosed
with cancer each year in the United States, and approximately half
a million cancer patients die annually despite the significant
progress made during the last decade in the diagnosis and treatment
of cancer. Leading types of cancer that affect a substantial number
of patients include colon cancer, breast cancer, prostate cancer,
and skin cancer. The need exists for improved drugs and therapeutic
methods for treating cancer.
[0005] Cardiovascular disorders that affect a significant number of
patients include atherosclerosis, arteriosclerosis, myocardial
infarction, angina pectoris, cardiac failure, embolism, thrombus,
and hypertension. Hypertension is particularly prevalent, with some
estimates suggesting approximately twenty-five percent of the adult
population worldwide being hypertensive. Although dietary and
lifestyle changes may reduce blood pressure, medications are often
necessary to reduce blood pressure to an acceptable level in
hypertensive patients. Examples of anti-hypertensive drugs include
angiotensin-converting enzyme (ACE) inhibitors, alpha blockers,
angiotensin II receptor antagonists, beta blockers, calcium channel
blockers, diuretics, and direct renin inhibitors. Without
treatment, hypertensive patients can have a significantly higher
risk of cardiovascular disorders and a reduced life expectancy. The
need exists for improved drugs and therapeutic methods for treating
cardiovascular disorders.
[0006] Also, current therapies for treating bacterial infections
are insufficient because many prominent, infection-causing
bacterial strains have developed resistance to current antibiotics.
Antibiotic resistance can result in severe adverse outcomes, such
as increased mortality, morbidity, and medical care costs for
patients suffering from common infections. Infections due to
organisms such as methicillin-resistant Staphylococcus aureus
(MRSA) occur with increasing frequency in hospitals and are
becoming more difficult to treat with conventional antibiotics. For
example, a recently discovered strain of Staphylococcus aureus was
resistant to treatment with vancomycin, a drug generally regarded
as a last line of defense against certain infections. Thus,
infection by antibiotic-resistant organisms is a significant health
threat for which new methods and compositions for treatment are
needed.
[0007] Another important medical therapy is blood transfusions.
Blood transfusions are a ubiquitious part of healthcare delivery.
In the United States (US), someone needs blood about every two
seconds and according to the 2009 National Blood Collection and
Utilization Survey Report (NBCUS), a total of 15 million units of
blood were transfused. Currently, blood products can be stored only
for short periods of time. Thus, one unmet medical need is for
compositions and methods capable of extending the storage life of
blood products.
[0008] Accordingly, there is need for new therapeutic methods and
compositions for extending the storage life of blood products,
enhancing the benefits of blood transfusions, and treating
disorders such as cancer, cardiovascular disorders, ischemic
conditions, and bacterial infections. The present invention
addresses these needs and provides other related advantages.
SUMMARY
[0009] The invention provides methods, compositions, and medical
kits comprising a nitrite-reductase promoter, such as an allosteric
modulator of hemoglobin, for use in treating medical disorders and
preservation of blood products. For example, in certain aspects,
the invention provides methods, compositions, and medical kits
comprising an inorganic nitrite salt and a nitrite-reductase
promoter, such as an allosteric modulator of hemoglobin, for use in
treating medical disorders, such as cancer, cardiovascular
disorders, ischemic conditions, hemolytic conditions, and bacterial
infections. In other aspects, the invention provides agents (e.g.,
allosteric modulator of hemoglobin) for treating a patient
suffering from reduced blood volume (e.g., a patient suffering from
hemorrhagic shock), performing a blood transfusion to a patient,
treating a patient suffering from anemia, and preserving an
isolated blood product. Various aspects and embodiments of the
invention are described in further detail below.
[0010] One aspect of the invention provides a method of treating or
preventing a disorder selected from the group consisting of cancer,
a cardiovascular disorder, an ischemic condition, a hemolytic
condition, or a bacterial infection. The method comprises
administering to a patient in need thereof a therapeutically
effective amount of (i) an inorganic nitrite salt, and (ii) an
allosteric modulator of hemoglobin that promotes nitrite reductase
activity. In certain embodiments, the allosteric modulator of
hemoglobin is a compound embraced by Formula I or Formula II,
wherein Formula I is represented by:
##STR00001##
or a pharmaceutically acceptable salt or solvate thereof, wherein
the variables are as defined in the detailed description, and
Formula II is represented by:
##STR00002##
or a pharmaceutically acceptable salt or solvate thereof, wherein
the variables are as defined in the detailed description.
[0011] Another aspect of the invention provides a method of
increasing the amount of nitric oxide produced by hemoglobin in a
patient. The method comprises administering to a patient in need
thereof a therapeutically effective amount of (i) an inorganic
nitrite salt, and (ii) an allosteric modulator of hemoglobin that
promotes nitrite reductase activity.
[0012] Another aspect of the invention provides a pharmaceutical
composition comprising (i) an inorganic nitrite salt, and (ii) an
allosteric modulator of hemoglobin that promotes nitrite reductase
activity. Still another aspect of the invention provides a kit for
treating a medical disorder. The kit comprises (i) an inorganic
nitrite salt, (ii) an allosteric modulator of hemoglobin, and (iii)
instructions for using the kit to treat a medical disorder.
[0013] Another aspect of the invention provides a method of
treating a patient suffering from reduced blood volume. The method
comprises administering to a patient in need thereof a blood
product by injection and a therapeutic agent selected from the
group consisting of an organonitro compound of Formula I,
organonitro compound of Formula II, hemoglobin conjugate of Formula
III, hemoglobin conjugate of Formula IV, and an erythrocyte cell
that has been exposed to an organonitro compound of Formula I or
II; wherein Formula I is represented by:
##STR00003##
or a pharmaceutically acceptable salt or solvate thereof, wherein
the variables are as defined in the detailed description, Formula
II is represented by:
##STR00004##
or a pharmaceutically acceptable salt or solvate thereof, wherein,
the variables are as defined in the detailed description, Formula
III is represented by:
##STR00005##
or a pharmaceutically acceptable salt or solvate thereof, wherein
the variables are as defined in the detailed description, and
Formula IV is represented by:
##STR00006##
or a pharmaceutically acceptable salt or solvate thereof, wherein
the variables are as defined in the detailed description.
[0014] Another aspect of the invention provides a method of
performing a blood transfusion to a patient. The method comprises
administering to a patient in need thereof a blood product by
injection and a therapeutic agent selected from the group
consisting of an organonitro compound of Formula I, organonitro
compound of Formula II, hemoglobin conjugate of Formula III,
hemoglobin conjugate of Formula IV, and an erythrocyte cell that
has been exposed to an organonitro compound of Formula I or II,
wherein Formula I is represented by:
##STR00007##
or a pharmaceutically acceptable salt or solvate thereof, wherein
the variables are as defined in the detailed description, Formula
II is represented by:
##STR00008##
or a pharmaceutically acceptable salt or solvate thereof: wherein
the variables are as defined in the detailed description, Formula
III is represented by:
##STR00009##
or a pharmaceutically acceptable salt or solvate thereof, wherein
the variables are as defined in the detailed description, and
Formula IV is represented by:
##STR00010##
or a pharmaceutically acceptable salt or solvate thereof, wherein
the variables are as defined in the detailed description.
[0015] Another aspect of the invention provides a method of
treating a patient suffering from anemia. The method comprises
administering to a patient in need thereof a therapeutic agent
selected from the group consisting of an organonitro compound of
Formula I, organonitro compound of Formula II, hemoglobin conjugate
of Formula III, hemoglobin conjugate of Formula IV, and an
erythrocyte cell that has been exposed to an organonitro compound
of Formula I or II; wherein Formula I is represented by:
##STR00011##
or a pharmaceutically acceptable salt or solvate thereof, wherein
the variables are as defined in the detailed description, Formula
II is represented by:
##STR00012##
or a pharmaceutically acceptable salt or solvate thereof, wherein
the variables are as defined in the detailed description, Formula
III is represented by:
##STR00013##
or a pharmaceutically acceptable salt or solvate thereof, wherein
the variables are as defined in the detailed description, and
Formula IV is represented by:
##STR00014##
or a pharmaceutically acceptable salt or solvate thereof, wherein
the variables are as defined in the detailed description.
[0016] Another aspect of the invention provides a method of
preserving an isolated blood product. The method comprises exposing
the isolated blood product to an agent selected from the group
consisting of an organonitro compound of Formula I, organonitro
compound of Formula II, hemoglobin conjugate of Formula III,
hemoglobin conjugate of Formula IV, and an erythrocyte cell that
has been exposed to an organonitro compound of Formula I or II,
wherein Formula I is represented by:
##STR00015##
or a pharmaceutically acceptable salt or solvate thereof, wherein
the variables are as defined in the detailed description, Formula
II is represented by:
##STR00016##
or a pharmaceutically acceptable salt or solvate thereof: wherein
the variables are as defined in the detailed description, Formula
III is represented by:
##STR00017##
or a pharmaceutically acceptable salt or solvate thereof, wherein
the variables are as defined in the detailed description, and
Formula IV is represented by:
##STR00018##
or a pharmaceutically acceptable salt or solvate thereof, wherein
the variables are as defined in the detailed description.
[0017] Another aspect of the invention provides an isolated blood
product composition. The composition comprises (i) a blood product,
and (ii) an agent selected from the group consisting of an
organonitro compound of Formula I, organonitro compound of Formula
II, hemoglobin conjugate of Formula III, hemoglobin conjugate of
Formula IV, and an erythrocyte cell that has been exposed to an
organonitro compound of Formula I or II; wherein Formula I is
represented by:
##STR00019##
or a pharmaceutically acceptable salt or solvate thereof, wherein
the variables are as defined in the detailed description, Formula
II is represented by:
##STR00020##
or a pharmaceutically acceptable salt or solvate thereof, wherein
the variables are as defined in the detailed description, Formula
III is represented by:
##STR00021##
or a pharmaceutically acceptable salt or solvate thereof, wherein
the variables are as defined in the detailed description, and
Formula IV is represented by:
##STR00022##
or a pharmaceutically acceptable salt or solvate thereof, wherein
the variables are as defined in the detailed description.
[0018] Another aspect of the invention provides an isolated
hemoglobin conjugate represented by Formula III or IV:
##STR00023##
or a pharmaceutically acceptable salt or solvate thereof, wherein
the variables are as defined in the detailed description, and
Formula IV is represented by:
##STR00024##
or a pharmaceutically acceptable salt or solvate thereof, wherein
the variables are as defined in the detailed description
[0019] Another aspect of the invention provides pharmaceutical
compositions comprising a pharmaceutically acceptable carrier and a
hemoglobin conjugate as defined in the detailed description, such
as a hemoglobin conjugate of Formula III or IV.
BRIEF DESCRIPTION OF THE FIGURES
[0020] FIG. 1 is a graph showing the cumulative amount of nitric
oxide formed from a blood sample over a thirty-minute time period
for multiple experiments (experimental conditions varied include
using air atmosphere, N.sub.2 atmosphere, and/or the presence or
absence of ABDNAZ), as described in Example 1;
[0021] FIG. 2 is a graph showing the cumulative amount of nitric
oxide formed from a blood sample over a thirty-minute time period
under an air atmosphere or N.sub.2 atmosphere (where d0 refers to
the first experiment, and dl refers to the second repetition of the
experiment), as described in Example 1;
[0022] FIG. 3 is a graph showing the cumulative amount of nitric
oxide formed from a blood sample mixed with ABDNAZ, where data is
shown for a thirty-minute time period under an air atmosphere or
N.sub.2 atmosphere (where d0 refers to the first experiment, and dl
refers to the second repetition of the experiment), as described in
Example 1;
[0023] FIG. 4 is a graph showing the cumulative amount of nitric
oxide formed from a blood sample under an atmosphere of air, where
the blood sample is optionally mixed with ABDNAZ (where d0 refers
to the first experiment, and dl refers to the second repetition of
the experiment), as described in Example 1;
[0024] FIG. 5 is a graph showing the cumulative amount of nitric
oxide formed from a blood sample under an atmosphere of N.sub.2,
where the blood sample is optionally mixed with ABDNAZ (where d0
refers to the first experiment, and dl refers to the second
repetition of the experiment), as described in Example 1;
[0025] FIG. 6 is a graph showing the amount of nitric oxide formed
in each three-minute period following the start of experiments for
multiple experiments (experimental conditions varied include using
an air atmosphere, N.sub.2 atmosphere, and/or the presence or
absence of ABDNAZ), as described in Example 1;
[0026] FIG. 7 is a graph showing the amount of nitric oxide formed
in each three-minute period following the start of experiments
where the blood sample is under an atmosphere of air or an
atmosphere of N.sub.2 as described in Example 1; and
[0027] FIG. 8 is a graph showing the amount of nitric oxide formed
in each three-minute period following the start of experiments
where the blood sample is mixed with ABDNAZ and is under an
atmosphere of air or an atmosphere of N.sub.2 as described in
Example 1.
[0028] FIG. 9 is a bar graph showing percent MetHb for the (i)
nitrite, (ii) RRx-001, and (iii) RRx-001+nitrite (RRx-001+N) groups
at 60 and 90 minutes post fluid resuscitation (FR), as further
described in Example 2. It is noted that the percent MetHb level in
normal, healthy individuals is about 1.
[0029] FIG. 10 depicts line graphs showing relative changes in
arteriolar diameter and blood flow during hemorrhagic shock and
resuscitation for all groups tested in Example 2.
[0030] FIG. 11 is a line graph showing relative changes in
functional capillary density (FCD) during hemorrhagic shock and
resuscitation for all groups tested in Example 2. Baseline averages
and standard deviations for each of the groups are: Sham,
106.+-.11; Blood, 107.+-.10; Nitrite, 107.+-.12; RRx-001, 112.+-.9;
RRx-001+Nitrite, 108.+-.9.
[0031] FIG. 12 is a line graph showing calculated vascular
resistance (MAP/blood flow) relative to baseline during hemorrhagic
shock and resuscitation for all groups tested in Example 2.
[0032] FIG. 13 is a bar graph showing the number of apoptotic and
necrotic cells at 8 hours following resuscitation for all groups
tested in Example 2. Data is presented as the average of
fluorescent cells counted in 40 selected visual fields
(210.times.160 .mu.m) for the tissue and the endothelial vessel
wall separately. **P<0.005 for both the number of apoptotic and
necrotic in the RRx-001 and RRx-001+nitrite groups compared to
blood only.
DETAILED DESCRIPTION OF THE INVENTION
[0033] The invention provides methods, compositions, and medical
kits comprising a nitrite-reductase promoter, such as an allosteric
modulator of hemoglobin, for use in treating medical disorders and
preservation of blood products. For example, in certain aspects,
the invention provides methods, compositions, and medical kits
comprising an inorganic nitrite salt and a nitrite-reductase
promoter, such as an allosteric modulator of hemoglobin, for use in
treating medical disorders, such as cancer, cardiovascular
disorders, ischemic conditions, hemolytic conditions, and bacterial
infections. In other aspects, the invention provides agents (e.g.,
allosteric modulator of hemoglobin) for treating a patient
suffering from reduced blood volume (e.g., a patient suffering from
hemorrhagic shock), performing a blood transfusion to a patient,
treating a patient suffering from anemia, and preserving an
isolated blood product. The practice of the present invention
employs, unless otherwise indicated, conventional techniques of
organic chemistry, pharmacology, cell biology, and biochemistry.
Such techniques are explained in the literature, such as in
"Comprehensive Organic Synthesis" (B. M. Trost & I. Fleming,
eds., 1991-1992); "Current protocols in molecular biology" (F. M.
Ausubel et al., eds., 1987, and periodic updates); and "Current
protocols in immunology" (J. E. Coligan et al., eds., 1991), each
of which is herein incorporated by reference in its entirety.
Various aspects of the invention are set forth below in sections;
however, aspects of the invention described in one particular
section are not to be limited to any particular section.
I. DEFINITIONS
[0034] To facilitate an understanding of the present invention, a
number of terms and phrases are defined below.
[0035] The terms "a" and "an" as used herein mean "one or more" and
include the plural unless the context is inappropriate.
[0036] The term "blood product" means (i) whole blood, or (ii)
component(s) isolated from whole.
[0037] The term "alkyl" as used herein refers to a saturated
straight or branched hydrocarbon, such as a straight or branched
group of 1-12, 1-10, or 1-6 carbon atoms, referred to herein as
C.sub.1-C.sub.12alkyl, C.sub.1-C.sub.10alkyl, and
C.sub.1-C.sub.6alkyl, respectively. Exemplary alkyl groups include,
but are not limited to, methyl, ethyl, propyl, isopropyl,
2-methyl-1-propyl, 2-methyl-2-propyl, 2-methyl-1-butyl,
3-methyl-1-butyl, 2-methyl-3-butyl, 2,2-dimethyl-1-propyl,
2-methyl-1-pentyl, 3-methyl-1-pentyl, 4-methyl-1-pentyl,
2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl,
2,2-dimethyl-1-butyl, 3,3-dimethyl-1-butyl, 2-ethyl-1-butyl, butyl,
isobutyl, t-butyl, pentyl, isopentyl, neopentyl, hexyl, heptyl,
octyl, etc.
[0038] The term "haloalkyl" refers to an alkyl group that is
substituted with at least one halogen. For example, --CH.sub.2F,
--CHF.sub.2, --CF.sub.3, --CH.sub.2CF.sub.3, --CF.sub.2CF.sub.3,
and the like.
[0039] The term "aralkyl" refers to an alkyl group substituted with
an aryl group.
[0040] The term "heteroaralkyl" refers to an alkyl group
substituted with a heteroaryl group.
[0041] The term "aryl" is art-recognized and refers to a
carbocyclic aromatic group. Representative aryl groups include
phenyl, naphthyl, anthracenyl, and the like. Unless specified
otherwise, the aromatic ring may be substituted at one or more ring
positions with, for example, halogen, azide, alkyl, aralkyl,
alkenyl, alkynyl, cycloalkyl, hydroxyl, alkoxyl, amino, nitro,
sulfhydryl, imino, amido, carboxylic acid, --C(O)alkyl,
--CO.sub.2alkyl, carbonyl, carboxyl, alkylthio, sulfonyl,
sulfonamido, sulfonamide, ketone, aldehyde, ester, heterocyclyl,
aryl or heteroaryl moieties, --CF.sub.3, --CN, or the like. The
term "aryl" also includes polycyclic ring systems having two or
more carbocyclic rings in which two or more carbons are common to
two adjoining rings (the rings are "fused rings") wherein at least
one of the rings is aromatic and, e.g., the other ring(s) may be
cycloalkyls, cycloalkenyls, cycloalkynyls, and/or aryls. In certain
embodiments, the aryl group is not substituted, i.e., it is
unsubstituted.
[0042] The "heteroaryl" is art-recognized and refers to aromatic
groups that include at least one ring heteroatom. In certain
instances, a heteroaryl group contains 1, 2, 3, or 4 ring
heteroatoms. Representative examples of heteroaryl groups includes
pyrrolyl, furanyl, thiophenyl, imidazolyl, oxazolyl, thiazolyl,
triazolyl, pyrazolyl, pyridinyl, pyrazinyl, pyridazinyl and
pyrimidinyl, and the like. Unless specified otherwise, the
heteroaryl ring may be substituted at one or more ring positions
with, for example, halogen, azide, alkyl, aralkyl, alkenyl,
alkynyl, cycloalkyl, hydroxyl, alkoxyl, amino, nitro, sulfhydryl,
imino, amido, carboxylic acid, --C(O)alkyl, --CO.sub.2alkyl,
carbonyl, carboxyl, alkylthio, sulfonyl, sulfonamido, sulfonamide,
ketone, aldehyde, ester, heterocyclyl, aryl or heteroaryl moieties,
--CF.sub.3, --CN, or the like. The term "heteroaryl" also includes
polycyclic ring systems having two or more rings in which two or
more carbons are common to two adjoining rings (the rings are
"fused rings") wherein at least one of the rings is heteroaromatic
and, e.g., the other ring(s) may be cycloalkyls, cycloalkenyls,
cycloalkynyls, and/or aryls. In certain embodiments, the heteroaryl
is a bicyclic aromatic ring in which both ring of the bicyclic
system are heteroaromatic. In certain embodiments, the heteroaryl
group is not substituted, i.e., it is unsubstituted.
[0043] The terms ortho, meta and para are art-recognized and refer
to 1,2-, 1,3- and 1,4-disubstituted benzenes, respectively. For
example, the names 1,2-dimethylbenzene and ortho-dimethylbenzene
are synonymous.
[0044] As used herein, the term "heterocyclic" represents, for
example, an aromatic or nonaromatic ring containing one or more
heteroatoms. The heteroatoms can be the same or different from each
other. Examples of heteroatoms include, but are not limited to
nitrogen, oxygen and sulfur. Aromatic and nonaromatic heterocyclic
rings are well-known in the art. Some nonlimiting examples of
aromatic heterocyclic rings include pyridine, pyrimidine, indole,
purine, quinoline and isoquinoline. Nonlimiting examples of
nonaromatic heterocyclic compounds include piperidine, piperazine,
morpholine, pyrrolidine and pyrazolidine. Examples of oxygen
containing heterocyclic rings include, but not limited to furan,
oxirane, 2H-pyran, 4H-pyran, 2H-chromene, and benzofuran. Examples
of sulfur-containing heterocyclic rings include, but are not
limited to, thiophene, benzothiophene, and parathiazine. Examples
of nitrogen containing rings include, but not limited to, pyrrole,
pyrrolidine, pyrazole, pyrazolidine, imidazole, imidazoline,
imidazolidine, pyridine, piperidine, pyrazine, piperazine,
pyrimidine, indole, purine, benzimidazole, quinoline, isoquinoline,
triazole, and triazine. Examples of heterocyclic rings containing
two different heteroatoms include, but are not limited to,
phenothiazine, morpholine, parathiazine, oxazine, oxazole,
thiazine, and thiazole. The heterocyclic ring is optionally further
substituted at one or more ring positions with, for example,
halogen, azide, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl,
hydroxyl, alkoxyl, amino, nitro, sulfhydryl, imino, amido,
carboxylic acid, --C(O)alkyl, --CO.sub.2alkyl, carbonyl, carboxyl,
alkylthio, sulfonyl, sulfonamido, sulfonamide, ketone, aldehyde,
ester, heterocyclyl, aryl or heteroaryl moieties, --CF.sub.3, --CN,
or the like.
[0045] The terms "amine" and "amino" are art-recognized and refer
to both unsubstituted and substituted amines, e.g., a moiety
represented by the general formula --N(R.sup.50)(R.sup.51), wherein
R.sup.50 and R.sup.51 each independently represent hydrogen, alkyl,
cycloalkyl, heterocyclyl, alkenyl, aryl, aralkyl, or
--(CH.sub.2).sub.m--R.sup.61; or R.sup.50 and R.sup.51, taken
together with the N atom to which they are attached complete a
heterocycle having from 4 to 8 atoms in the ring structure;
R.sup.61 represents an aryl, a cycloalkyl, a cycloalkenyl, a
heterocycle or a polycycle; and m is zero or an integer in the
range of 1 to 8. In embodiments, R.sup.50 and R.sup.51 each
independently represent hydrogen, alkyl, alkenyl, or
--(CH.sub.2).sub.m--R.sup.61.
[0046] The terms "alkoxyl" or "alkoxy" are art-recognized and refer
to an alkyl group, as defined above, having an oxygen radical
attached thereto. Representative alkoxyl groups include methoxy,
ethoxy, propyloxy, tert-butoxy and the like. An "ether" is two
hydrocarbons covalently linked by an oxygen. Accordingly, the
substituent of an alkyl that renders that alkyl an ether is or
resembles an alkoxyl, such as may be represented by one of
--O-alkyl, --O-alkenyl, --O-alkynyl,
--O--(CH.sub.2).sub.m--R.sub.61, where m and R.sub.61 are described
above.
[0047] The terms "ABDNAZ" and "RRx-001" are used interchangeably
and refer to the compound having the following structure:
##STR00025##
[0048] Certain compounds contained in compositions of the present
invention may exist in particular geometric or stereoisomeric
forms. The present invention contemplates all such compounds,
including cis- and trans-isomers, R- and S-enantiomers,
diastereomers, (D)-isomers, (L)-isomers, the racemic mixtures
thereof, and other mixtures thereof, as falling within the scope of
the invention. Additional asymmetric carbon atoms may be present in
a substituent such as an alkyl group. All such isomers, as well as
mixtures thereof, are intended to be included in this
invention.
[0049] If, for instance, a particular enantiomer of a compound of
the present invention is desired, it may be prepared by asymmetric
synthesis, or by derivation with a chiral auxiliary, where the
resulting diastereomeric mixture is separated and the auxiliary
group cleaved to provide the pure desired enantiomers.
Alternatively, where the molecule contains a basic functional
group, such as amino, or an acidic functional group, such as
carboxyl, diastereomeric salts are formed with an appropriate
optically-active acid or base, followed by resolution of the
diastereomers thus formed by fractional crystallization or
chromatographic means well known in the art, and subsequent
recovery of the pure enantiomers.
[0050] As used herein, the terms "subject" and "patient" refer to
organisms to be treated by the methods of the present invention.
Such organisms are preferably mammals (e.g., murines, simians,
equines, bovines, porcines, canines, felines, and the like), and
more preferably humans. The term "non-anemic patient" refers to a
patient that does not suffer from anemia.
[0051] As used herein, the term "effective amount" refers to the
amount of a compound (e.g., a compound of the present invention)
sufficient to effect beneficial or desired results. An effective
amount can be administered in one or more administrations,
applications or dosages and is not intended to be limited to a
particular formulation or administration route. As used herein, the
term "treating" includes any effect, e.g., lessening, reducing,
modulating, ameliorating or eliminating, that results in the
improvement of the condition, disease, disorder, and the like, or
ameliorating a symptom thereof.
[0052] As used herein, the term "pharmaceutical composition" refers
to the combination of an active agent with a carrier, inert or
active, making the composition especially suitable for diagnostic
or therapeutic use in vivo or ex vivo.
[0053] As used herein, the term "pharmaceutically acceptable
carrier" refers to any of the standard pharmaceutical carriers,
such as a phosphate buffered saline solution, water, emulsions
(e.g., such as an oil/water or water/oil emulsions), and various
types of wetting agents. The compositions also can include
stabilizers and preservatives. For examples of carriers,
stabilizers and adjuvants. (See e.g., Martin, Remington's
Pharmaceutical Sciences, 15th Ed., Mack Publ. Co., Easton, Pa.
[1975]).
[0054] As used herein, the term "pharmaceutically acceptable salt"
refers to any pharmaceutically acceptable salt (e.g., acid or base)
of a compound of the present invention which, upon administration
to a subject, is capable of providing a compound of this invention
or an active metabolite or residue thereof. As is known to those of
skill in the art, "salts" of the compounds of the present invention
may be derived from inorganic or organic acids and bases. Examples
of acids include, but are not limited to, hydrochloric,
hydrobromic, sulfuric, nitric, perchloric, fumaric, maleic,
phosphoric, glycolic, lactic, salicylic, succinic,
toluene-p-sulfonic, tartaric, acetic, citric, methanesulfonic,
ethanesulfonic, formic, benzoic, malonic, naphthalene-2-sulfonic,
benzenesulfonic acid, and the like. Other acids, such as oxalic,
while not in themselves pharmaceutically acceptable, may be
employed in the preparation of salts useful as intermediates in
obtaining the compounds of the invention and their pharmaceutically
acceptable acid addition salts.
[0055] Examples of bases include, but are not limited to, alkali
metals (e.g., sodium) hydroxides, alkaline earth metals (e.g.,
magnesium), hydroxides, ammonia, and compounds of formula
NW.sub.4.sup.+, wherein W is C.sub.14 alkyl, and the like.
[0056] Examples of salts include, but are not limited to: acetate,
adipate, alginate, aspartate, benzoate, benzenesulfonate,
bisulfate, butyrate, citrate, camphorate, camphorsulfonate,
cyclopentanepropionate, digluconate, dodecylsulfate,
ethanesulfonate, fumarate, flucoheptanoate, glycerophosphate,
hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide,
hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate,
methanesulfonate, 2-naphthalenesulfonate, nicotinate, oxalate,
palmoate, pectinate, persulfate, phenylpropionate, picrate,
pivalate, propionate, succinate, tartrate, thiocyanate, tosylate,
undecanoate, and the like. Other examples of salts include anions
of the compounds of the present invention compounded with a
suitable cation such as Na.sup.+, NH.sub.4.sup.+, and
NW.sub.4.sup.+ (wherein W is a C.sub.14 alkyl group), and the
like.
[0057] For therapeutic use, salts of the compounds of the present
invention are contemplated as being pharmaceutically acceptable.
However, salts of acids and bases that are non-pharmaceutically
acceptable may also find use, for example, in the preparation or
purification of a pharmaceutically acceptable compound.
[0058] The term "isolated" refers to material that is removed from
its original environment (e.g., the natural environment if it is
naturally occurring).
[0059] Throughout the description, where compositions and kits are
described as having, including, or comprising specific components,
or where processes and methods are described as having, including,
or comprising specific steps, it is contemplated that,
additionally, there are compositions and kits of the present
invention that consist essentially of, or consist of, the recited
components, and that there are processes and methods according to
the present invention that consist essentially of, or consist of,
the recited processing steps.
[0060] As a general matter, compositions specifying a percentage
are by weight unless otherwise specified. Further, if a variable is
not accompanied by a definition, then the previous definition of
the variable controls.
II. COMBINATION THERAPY OF INORGANIC NITRITE SALT AND
NITRITE-REDUCTASE PROMOTER
[0061] The invention provides combination therapy using an
inorganic nitrite salt in combination with a nitrite-reductase
promoter. Exemplary inorganic nitrite salts and exemplary
nitrite-reductase promoters for use in the combination therapy
methods, pharmaceutical compositions, and medical kits are
described below. In addition, because the combination therapy may
optionally comprise administration of one or more additional
therapeutic agents for treatment of the designated medical
disorder, exemplary additional therapeutic agents for treating
exemplary medical disorders are described below.
[0062] A. Inorganic Nitrite Salts
[0063] The inorganic nitrite salt may be an alkali metal nitrite
salt, an alkaline earth metal nitrite salt, or ammonium nitrite
salt. Exemplary alkali metal nitrite salts include sodium nitrite,
postassium nitrite, lithium nitrite, cesium nitrite, and rubidium
nitrite. Exemplary alkaline earth metal nitrite salts include
magnesium nitrite, calcium nitrite, barium nitrite, and strontium
nitrite. Additional exemplary metal-based inorganic nitrite salts
include silver (I) nitrite (AgNO.sub.2), cobalt(II) nitrite
(Co(NO.sub.2).sub.2), and zinc nitrite (Zn(NO.sub.2).sub.2). The
alkali metal nitrite salt, alkaline earth metal nitrite salt, or
ammonium nitrite salt may be in the form of solvate, such as a
hydrate (e.g., a mono-hydrate or dehydrate). Alternatively, the
alkali metal nitrite salt, alkaline earth metal nitrite salt, or
ammonium nitrite salt may be anhydrous.
[0064] Exemplary ammonium nitrite salts include compounds embraced
by the formula NO.sub.2--N(R').sub.4, wherein R' represents
independently for each occurrence hydrogen, optionally substituted
alkyl, optionally substituted heteroalkyl, optionally substituted
cycloalkyl, optionally substituted heterocycloalkyl, optionally
substituted aryl, optionally substituted aralkyl, optionally
substituted heteroaryl, or optionally substituted heteroaralkyl. In
certain other embodiments, the ammonium nitrite salt is arginine
nitrite, ammonium nitrite (NH.sub.4NO.sub.2), or
tetramethylammonium nitrite.
[0065] B. Nitrite-Reductase Promoters
[0066] The nitrite reductase promoter enhances conversion of
nitrite to nitric oxide in vivo. One exemplary class of
nitrite-reductase promoters is an allosteric modulator of
hemoglobin, such as compounds that bind to the beta-cysteine-93
residue of hemoglobin to enhance the nitrite-reductase activity of
hemoglobin. Another exemplary class of nitrite-reductase promoters
is an agent that modulates the oxygen binding affinity of
hemoglobin and/or erythrocyte cells, such as an agent that
increases oxygen binding affinity of hemoglobin and/or erythrocyte
cells. Co-administration of a nitrite reductase promoter with an
inorganic nitrite salt results in increased levels of nitric oxide
in vivo. One benefit of the combination therapy is that the nitrite
reductase promoter allows for generation of beneficial levels of
nitric oxide in vivo, while minimizing the amount of inorganic
nitrite salt that must be administered to the patient.
[0067] Exemplary allosteric modulators of hemoglobin contemplated
for use in the methods, compositions, and kits include nitrosating
agents such as S-nitroso-N-acetylcysteine,
S-nitrosocysteinylglycine, S-nitrosocysteine,
S-nitrosohomocysteine, metal nitrosyl complexes, S-nitro compounds,
S-nitroso compounds, thionitrites, diazeniumdiolates, and other
related nitrosating agents as described in Feelisch, M. and
Stamler, J. S., "Donors of Nitrogen Oxides" chapter 7, pp. 71-115
in Methods in Nitric Oxide Research (Freelisch, M. and Stamler, J.
S., eds.) John Wiley and Sons, Ltd., Chichester, U.K. (1996), the
contents of which are hereby incorporated by reference in their
entirety. A nitrosating agent can be chosen for minimal oxidation
of the heme iron of hemoglobin, and maximum activity in
nitosylating thiol groups such as found on cysteine. Other
exemplary allosteric modulators of hemoglobin contemplated for use
in the methods, compositions, and kits include 4-pyridylmethyl
chloride, an alkoxyalkylchloride, dimethoxymethane,
N-(hydroxymethyl)acetamide, triphenylmethyl chloride, acetyl
chloride, 2-chloroacetic acid, acetic anhydride, a haloacetamide
(such as, iodoacetamide, bromoacetamide, chloroacetamide, or
fluoroacetamide), a haloacetate (such as iodoacetate, bromoacetate,
chloroacetate, or fluoroacetate), benzyl chloride, benzoyl
chloride, di-tert-butyl dicarbonate, p-hydroxyphenacyl bromide,
p-acetoxybenzyl chloride, p-methoxybenzyl chloride,
2,4-dinitrophenyl fluoride, tetrahydropyran,
acetamidohydroxymethane, acetone, bis-carboethoxyethene,
2,2,2-trichloroethoxycarbonyl chloride, tert-butoxycarbonyl
chloride, an alkyl isocyanate, and an alkoxyalkyl isocyanate. In
certain other embodiments, the allosteric modulator of hemoglobin
is an optionally substituted alkyl-R*, optionally substituted
aralkyl-R*, or optionally substituted heteroaralkyl-R*, wherein R*
is a leaving group, such as halogen, an alkyl sulfonate,
arylsulfonate, alkyl acetate, or haloalkyl acetate. In certain
other embodiments, the allosteric modulator of hemoglobin is an
optionally substituted alkyl-NCO, optionally substituted aryl-NCO,
optionally substituted aralkyl-NCO, optionally substituted
heterocycyl-NCO, optionally substituted heteroaryl-NCO, or
optionally substituted heteroaralkyl-NCO. In certain other
embodiments, the allosteric modulator of hemoglobin is an
optionally substituted alkyl-C(O)X, optionally substituted
aryl-C(O)X, optionally substituted aralkyl-C(O)X, optionally
substituted heterocycyl-C(O)X, optionally substituted
heteroaryl-C(O)X, or optionally substituted heteroaralkyl-C(O)X,
where X is a leaving group, such as halogen or --OC(O)alkyl.
[0068] In certain embodiments, the sulfhydryl of the
.beta.93-cysteine on hemoglobin may be alkylated with an allosteric
modulator of hemoglobin that is a derivatized dextran. For example,
in certain embodiments, the dextran may be derivatized to contain a
free amino group (e.g., using cyanogen bromide and diaminoethane),
and the free amino group may be acylated with an alkylating moiety
(e.g., bromoacetyl bromide) that can alkylate the sulfhydryl of the
.beta.93-cysteine.
[0069] In certain other embodiments, the allosteric modulator of
hemoglobin is a polyalkylene glycol. Polyalkylene glycols
containing a reactive group are contemplated to react with the
.beta.93-cysteine residue of hemoglobin to modulate hemoglobin
activity. In certain embodiments, the polyalkylene glycol contains
a maleimide group, such as (polyethylene glycol)-maleimide. In
certain other embodiments, the polyalkylene glycol contains a
N-hydroxysuccinimide group. The polyethylene glycol may have a
weight average molecular weight of about 200 g/mol to about 100,000
g/mol, about 200 g/mol to about 20,000 g/mol, about 200 g/mol to
about 1,000 g/mol, or about 1,000 g/mol to about 10,000 g/mol.
[0070] In certain other embodiments, the allosteric modulator of
hemoglobin is an organonitro compound embraced by Formula I:
##STR00026##
[0071] or a pharmaceutically acceptable salt or solvate thereof,
wherein:
[0072] A.sup.1 is --C(O)-- or
--(C(R.sup.3).sub.2).sub.xC(O)(C(R.sup.3).sub.2).sub.x--;
[0073] A.sup.2 is N or --C(R.sup.4)--;
[0074] R.sup.1 is halogen, --OS(O).sub.2R.sup.5, or
--OC(O)CF.sub.3;
[0075] R.sup.2 is C.sub.1-C.sub.6alkyl;
[0076] R.sup.3 and R.sup.4 each represent independently for each
occurrence hydrogen or C.sub.1-C.sub.5alkyl;
[0077] R.sup.5 is C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6haloalkyl,
aryl, or aralkyl;
[0078] m and p are independently 1, 2, or 3; and
[0079] n and x each represent independently for each occurrence 0,
1, 2, or 3.
[0080] In certain embodiments, the allosteric modulator of
hemoglobin is an organonitro compound embraced by Formula I as
defined by particular definitions for variables in Formula I, such
as where A.sup.1 is --C(O)--. In certain other embodiments, A.sup.1
is --(C(R.sup.3).sub.2).sub.xC(O)(C(R.sup.3).sub.2).sub.x--. In
certain other embodiments, A.sup.1 is
--C(O)(C(R.sup.3).sub.2).sub.x--.
[0081] In certain embodiments, A.sup.2 is N. In certain other
embodiments, A.sup.2 is --C(R.sup.4)--.
[0082] In certain embodiments, R.sup.1 is halogen,
--OS(O).sub.2R.sup.5, or --OC(O)CF.sub.3. In certain other
embodiments, R.sup.1 is halogen. In certain other embodiments,
R.sup.1 is --OS(O).sub.2R.sup.5. In certain other embodiments,
R.sup.1 is --OC(O)CF.sub.3. In certain other embodiments, R.sup.1
is chloro, bromo, --OS(O).sub.2-(para-methylphenyl),
--OS(O).sub.2CH.sub.3, --OS(O).sub.2CF.sub.3, or --OC(O)CF.sub.3.
In certain embodiments, R.sup.1 is bromo.
[0083] In certain embodiments, m is 2. In certain other
embodiments, m is 1.
[0084] In certain embodiments, n is 0. In certain other
embodiments, n is 1. In certain other embodiments, n is 2.
[0085] In certain embodiments, p is 1. In certain other
embodiments, p is 2. In certain other embodiments, p is 3.
[0086] The description above describes multiple embodiments
relating to compounds of Formula I. The patent application
specifically contemplates all combinations of the embodiments. For
example, the invention contemplates a compound of Formula I wherein
A.sup.1 is --C(O)--, A.sup.2 is N, R.sup.1 is halogen, and n is
0.
[0087] In certain embodiments, the compound is a compound of
Formula I-A:
##STR00027##
[0088] or a pharmaceutically acceptable salt or solvate thereof,
wherein:
[0089] A is N or C(H);
[0090] R.sup.1 is chloro, bromo,
--OS(O).sub.2--(C.sub.1-C.sub.6alkyl),
--OS(O).sub.2--(C.sub.1-C.sub.6haloalkyl),
--OS(O).sub.2-(para-methylphenyl), or --OC(O)CF.sub.3;
[0091] R.sup.2 represents independently for each occurrence
hydrogen or methyl; and
[0092] y represents independently for each occurrence 1 or 2.
[0093] In certain embodiments, the allosteric modulator of
hemoglobin is an organonitro compound embraced by Formula I-A as
defined by particular definitions for variables in Formula I-A,
such as where A is N. In certain other embodiments, A is C(H).
[0094] In certain embodiments, R.sup.1 is chloro or bromo. In
certain embodiments, R.sup.1 is chloro. In certain other
embodiments, R.sup.1 is bromo. In certain embodiments, R.sup.1 is
--OS(O).sub.2--(C.sub.1-C.sub.6alkyl),
--OS(O).sub.2--(C.sub.1-C.sub.6haloalkyl), or
--OS(O).sub.2-(para-methylphenyl). In certain other embodiments,
R.sup.1 is --OS(O).sub.2CH.sub.3, --OS(O).sub.2CF.sub.3, or
--OS(O).sub.2-(para-methylphenyl). In certain other embodiments,
R.sup.1 is --OC(O)CF.sub.3.
[0095] In certain embodiments, R.sup.2 is hydrogen or methyl. In
certain embodiments, R2 is hydrogen.
[0096] In certain embodiments, y is 1. In certain embodiments, one
occurrence of y is 1, and the other occurrence of y is 2. In
certain other embodiments, y is 2.
[0097] The description above describes multiple embodiments
relating to compounds of Formula I-A. The patent application
specifically contemplates all combinations of the embodiments. For
example, the invention contemplates a compound of Formula I-A
wherein A is N, R.sup.1 is chloro or bromo, and R.sup.2 is
hydrogen.
[0098] In certain embodiments, the compound is
##STR00028##
or a pharmaceutically acceptable salt or solvate thereof. In
certain other embodiments, the compound is
##STR00029##
[0099] In certain other embodiments, the allosteric modulator of
hemoglobin is an organonitro compound embraced by Formula II:
##STR00030##
[0100] or a pharmaceutically acceptable salt or solvate thereof:
wherein:
[0101] A.sup.1 is --C(O)-- or
--(C(R.sup.5).sub.2).sub.xC(O)(C(R.sup.5).sub.2).sub.x--;
[0102] A.sup.2 is --N(R.sup.5)-- or --C(R.sup.2)(R.sup.3)--;
[0103] R.sup.1 is halogen, --OS(O).sub.2R.sup.6, or
--OC(O)CF.sub.3;
[0104] R.sup.2 and R.sup.3 each represent independently for each
occurrence hydrogen or C.sub.1-C.sub.6alkyl; or R.sup.2 and R.sup.3
are taken together with the carbon atom to which they are attached
to form a 3-6 membered, saturated carbocyclic ring;
[0105] R.sup.4 is hydrogen or C.sub.1-C.sub.6alkyl;
[0106] R.sup.5 represents independently for each occurrence
hydrogen or C.sub.1-C.sub.6alkyl;
[0107] R.sup.6 is C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6haloalkyl,
aryl, or aralkyl;
[0108] t is an integer in the range from 1 to 12; and
[0109] x represents independently for each occurrence 0, 1, 2, or
3.
[0110] In certain embodiments, the allosteric modulator of
hemoglobin is an organonitro compound embraced by Formula II as
defined by particular definitions for variables in Formula II, such
as where A.sup.1 is --C(O)--. In certain other embodiments, A.sup.1
is --(C(R.sup.5).sub.2).sub.xC(O)(C(R.sup.5).sub.2).sub.x--. In
certain other embodiments, A.sup.1 is
--C(O)(C(R.sup.5).sub.2).sub.x--.
[0111] In certain embodiments, A.sup.2 is --N(R.sup.5)--. In
certain other embodiments, A.sup.2 is --C(R.sup.2)(R.sup.3)--.
[0112] In certain embodiments, R.sup.1 is halogen. In certain other
embodiments, R.sup.1 is --OS(O).sub.2R.sup.6. In certain other
embodiments, R.sup.1 is --OC(O)CF.sub.3. In certain other
embodiments, R.sup.1 is chloro, bromo,
--OS(O).sub.2-(para-methylphenyl), --OS(O).sub.2CH.sub.3,
--OS(O).sub.2CF.sub.3, or --OC(O)CF.sub.3. In certain embodiments,
R.sup.1 is bromo.
[0113] In certain embodiments, R.sup.2 and R.sup.3 each represent
independently for each occurrence hydrogen or C.sub.1-C.sub.6alkyl.
In certain other embodiments, R.sup.2 and R.sup.3 each represent
independently for each occurrence hydrogen, methyl, ethyl, or
propyl. In certain other embodiments, R.sup.2 and R.sup.3 each
represent independently for each occurrence hydrogen or methyl. In
certain embodiments, R.sup.2 and R.sup.3 are hydrogen.
[0114] In certain embodiments, R.sup.4 is hydrogen, methyl, ethyl,
propyl, butyl, or pentyl. In certain other embodiments, R.sup.4 is
methyl, ethyl or propyl. In certain other embodiments, R.sup.4 is
methyl.
[0115] In certain embodiments, R.sup.5 is hydrogen or methyl. In
certain other embodiments, R.sup.5 is hydrogen.
[0116] In certain embodiments, R.sup.6 is C.sub.1-C.sub.6alkyl or
C.sub.1-C.sub.6haloalkyl. In certain other embodiments, R.sup.6 is
methyl, ethyl, or trifluoromethyl. In certain other embodiments,
R.sup.6 is aryl, such as phenyl.
[0117] In certain embodiments, t is 1, 2, 3, 4, 5 or 6. In certain
other embodiments, t is 1, 2, or 3. In certain other embodiments, t
is 1. In certain embodiments, x is 1 or 2.
[0118] The description above describes multiple embodiments
relating to compounds of Formula II. The patent application
specifically contemplates all combinations of the embodiments. For
example, the invention contemplates a compound of Formula II
wherein A.sup.1 is --C(O)--, A.sup.2 is --N(R.sup.5)--, and R.sup.2
and R.sup.3 are hydrogen.
[0119] In certain embodiments, the compound is a compound of
Formula II-A:
##STR00031##
[0120] or a pharmaceutically acceptable salt or solvate thereof:
wherein:
[0121] A is --N(R.sup.5)-- or --C(R.sup.2)(R.sup.3)--;
[0122] R.sup.1 is chloro, bromo,
--OS(O).sub.2--(C.sub.1-C.sub.6alkyl),
--OS(O).sub.2--(C.sub.1-C.sub.6haloalkyl),
--OS(O).sub.2-(para-methylphenyl), or --OC(O)CF.sub.3;
[0123] R.sup.2, R.sup.3, and R.sup.5 each represent independently
for each occurrence hydrogen or methyl;
[0124] R.sup.4 is hydrogen or C.sub.1-C.sub.6alkyl; and
[0125] t is 1, 2, or 3.
[0126] In certain embodiments, the allosteric modulator of
hemoglobin is an organonitro compound embraced by Formula II-A as
defined by particular definitions for variables in Formula II-A,
such as where A is --N(R.sup.5)--. In certain other embodiments, A
is --N(CH.sub.3)--. In certain other embodiments, A is
--C(R.sup.2)(R.sup.3)--. In certain other embodiments, A is
--CH.sub.2--.
[0127] In certain embodiments, R.sup.1 is chloro. In certain other
embodiments, R.sup.1 is bromo. In certain embodiments, R.sup.1 is
--OS(O).sub.2--(C.sub.1-C.sub.6alkyl),
--OS(O).sub.2--(C.sub.1-C.sub.6haloalkyl), or
--OS(O).sub.2-(para-methylphenyl). In certain other embodiments,
R.sup.1 is --OS(O).sub.2CH.sub.3, --OS(O).sub.2CF.sub.3, or
--OS(O).sub.2-(para-methylphenyl). In certain other embodiments,
R.sup.1 is --OC(O)CF.sub.3.
[0128] In certain embodiments, R.sup.2 and R.sup.3 are
hydrogen.
[0129] In certain embodiments, R.sup.4 is hydrogen, methyl, ethyl,
propyl, butyl, or pentyl. In certain other embodiments, R.sup.4 is
methyl, ethyl or propyl. In certain other embodiments, R.sup.4 is
methyl.
[0130] In certain embodiments, R.sup.5 is hydrogen or methyl. In
certain other embodiments, R.sup.5 is hydrogen.
[0131] The description above describes multiple embodiments
relating to compounds of Formula II-A. The patent application
specifically contemplates all combinations of the embodiments. For
example, the invention contemplates a compound of Formula II-A
wherein A is --N(R.sup.5)--, and R.sup.2 and R.sup.3 are
hydrogen.
[0132] In certain other embodiments, the allosteric modulator of
hemoglobin is one of the compounds listed in Tables 1, 2, or 3
below or a pharmaceutically acceptable salt or solvate thereof
TABLE-US-00001 TABLE 1 ##STR00032## Com- pound No. X Y I-1 Br
##STR00033## I-2 Br ##STR00034## I-3 Br ##STR00035## I-4 Br
##STR00036## I-5 Br ##STR00037## I-6 Br ##STR00038## I-7 Br
##STR00039## I-8 Br ##STR00040## I-9 Br ##STR00041## I-10 Br
##STR00042## I-11 Br ##STR00043## I-12 Br ##STR00044## I-13 Br
##STR00045## I-14 Br ##STR00046## I-15 Br ##STR00047## I-16 Br
##STR00048## I-17 Br ##STR00049## I-18 Br ##STR00050## I-19 Cl
##STR00051## I-20 Cl ##STR00052## I-21 Cl ##STR00053## I-22 Cl
##STR00054## I-23 I ##STR00055## I-24 I ##STR00056## I-25 I
##STR00057## I-26 I ##STR00058## I-27 --OS(O).sub.2CH.sub.3
##STR00059## I-28 --OS(O).sub.2CH.sub.3 ##STR00060## I-29
--OS(O).sub.2CF.sub.3 ##STR00061## I-30 --OS(O).sub.2CF.sub.3
##STR00062## I-31 ##STR00063## ##STR00064## I-32 ##STR00065##
##STR00066## I-33 --OC(O)CF.sub.3 ##STR00067## I-34 --OC(O)CF.sub.3
##STR00068## I-35 --OC(O)CF.sub.3 ##STR00069##
TABLE-US-00002 TABLE 2 ##STR00070## Compound No. X A Y II-1 Br
--CH.sub.2C(O)-- ##STR00071## II-2 Br --CH.sub.2C(O)-- ##STR00072##
II-3 Br --CH.sub.2C(O)-- ##STR00073## II-4 Br --CH.sub.2C(O)--
##STR00074## II-5 Br --CH.sub.2C(O)-- ##STR00075## II-6 Br
--CH.sub.2C(O)-- ##STR00076## II-7 Br --CH.sub.2C(O)-- ##STR00077##
II-8 Br --C(O)CH.sub.2CH.sub.2-- ##STR00078## II-9 Br
--C(O)CH.sub.2CH.sub.2-- ##STR00079## II-10 Br
--C(O)CH.sub.2CH.sub.2-- ##STR00080## II-11 Br
--C(O)CH.sub.2CH.sub.2-- ##STR00081## II-12 Br --CH.sub.2--
##STR00082## II-13 Br --CH.sub.2-- ##STR00083## II-14 Br
--CH.sub.2-- ##STR00084## II-15 Br --CH.sub.2-- ##STR00085## II-16
Br --CH.sub.2C(O)-- ##STR00086## II-17 Br --CH.sub.2C(O)--
##STR00087## II-18 Br --C(O)CH.sub.2CH.sub.2-- ##STR00088## II-19
Br --C(O)CH.sub.2CH.sub.2-- ##STR00089## II-20 Br --CH.sub.2--
##STR00090## II-21 Br --CH.sub.2-- ##STR00091## II-22 Br
--CH.sub.2-- ##STR00092## II-23 Cl --CH.sub.2C(O)-- ##STR00093##
II-24 Cl --C(O)CH.sub.2CH.sub.2-- ##STR00094## II-25 Cl
--CH.sub.2-- ##STR00095## II-26 Cl --CH.sub.2C(O)-- ##STR00096##
II-27 Cl --C(O)CH.sub.2CH.sub.2-- ##STR00097## II-28 Cl
--CH.sub.2-- ##STR00098## II-29 --OS(O).sub.2CH.sub.3
--CH.sub.2C(O)-- ##STR00099## II-30 --OS(O).sub.2CH.sub.3
--CH.sub.2C(O)-- ##STR00100## II-31 --OS(O).sub.2CH.sub.3
--C(O)CH.sub.2CH.sub.2-- ##STR00101## II-32 --OS(O).sub.2CH.sub.3
--CH.sub.2-- ##STR00102## II-33 --OS(O).sub.2CF.sub.3
--CH.sub.2C(O)-- ##STR00103## II-34 --OS(O).sub.2CF.sub.3
--CH.sub.2C(O)-- ##STR00104## II-35 --OS(O).sub.2CF.sub.3
--C(O)CH.sub.2CH.sub.2-- ##STR00105## II-36 --OS(O).sub.2CF.sub.3
--CH.sub.2-- ##STR00106## II-37 ##STR00107## --CH.sub.2C(O)--
##STR00108## II-38 ##STR00109## --CH.sub.2C(O)-- ##STR00110## II-39
##STR00111## --C(O)CH.sub.2CH.sub.2-- ##STR00112## II-40
##STR00113## --CH.sub.2-- ##STR00114## II-41 --OC(O)CF.sub.3
--CH.sub.2C(O)-- ##STR00115## II-42 --OC(O)CF.sub.3
--CH.sub.2C(O)-- ##STR00116## II-43 --OC(O)CF.sub.3
--C(O)CH.sub.2CH.sub.2-- ##STR00117## II-44 --OC(O)CF.sub.3
--CH.sub.2-- ##STR00118##
TABLE-US-00003 TABLE 3 ##STR00119## Compound No. X Y Z III-1 Br
--N(H)CH.sub.2-- methyl III-2 Br --N(H)CH.sub.2-- ethyl III-3 Br
--N(H)CH.sub.2-- n-pentyl III-4 Br --N(H)CH.sub.2-- hydrogen III-5
Br --N(H)CH.sub.2CH.sub.2-- methyl III-6 Br
--N(H)(CH.sub.2).sub.4-- methyl III-7 Br --N(CH.sub.3)CH.sub.2--
methyl III-8 Br --N(CH.sub.3)(CH.sub.2).sub.3-- methyl III-9 Br
--N(H)C(CH.sub.3)(H)-- methyl III-10 Br
--N(H)C(CH.sub.3)(H)CH.sub.2-- methyl III-11 Br --CH.sub.2-- methyl
III-12 Br --(CH.sub.2).sub.2-- methyl III-13 Br --CH.sub.2-- ethyl
III-14 Br --(CH.sub.2).sub.4-- isopropyl III-15 Br
--(CH.sub.2).sub.2-- n-pentyl III-16 Br --CH.sub.2-- hydrogen
III-17 Br --CH.sub.2CH.sub.2C(CH.sub.3).sub.2-- methyl III-18 Br
--CH.sub.2C(CH.sub.3).sub.2CH.sub.2-- methyl III-19 Cl
--N(H)CH.sub.2-- methyl III-20 Cl --N(H)CH.sub.2-- ethyl III-21 Cl
--N(CH.sub.3)CH.sub.2-- methyl III-22 Cl --(CH.sub.2).sub.2--
methyl III-23 I --N(H)CH.sub.2-- methyl III-24 I --N(H)CH.sub.2--
ethyl III-25 I --N(CH.sub.3)CH.sub.2-- methyl III-26 I
--(CH.sub.2).sub.2-- methyl III-27 --OS(O).sub.2CH.sub.3
--N(H)CH.sub.2-- methyl III-28 --OS(O).sub.2CH.sub.3
--N(CH.sub.3)CH.sub.2-- methyl III-29 --OS(O).sub.2CF.sub.3
--N(H)CH.sub.2-- methyl III-30 --OS(O).sub.2CF.sub.3
--N(CH.sub.3)CH.sub.2-- methyl III-31 ##STR00120## --N(H)CH.sub.2--
methyl III-32 ##STR00121## --N(CH.sub.3)CH.sub.2-- methyl III-33
--OC(O)CF.sub.3 --N(H)CH.sub.2-- methyl III-34 --OC(O)CF.sub.3
--N(CH.sub.3)CH.sub.2-- methyl III-35 --OC(O)CF.sub.3
--(CH.sub.2).sub.2-- methyl
[0133] Methods for preparing compounds described herein are
illustrated in the following synthetic schemes. These schemes are
given for the purpose of illustrating the invention, and should not
be regarded in any manner as limiting the scope or the spirit of
the invention. Starting materials shown in the schemes can be
obtained from commercial sources or can be prepared based on
procedures described in the literature.
[0134] The synthetic route illustrated in Scheme 1 depicts a
general method for preparing cyclic geminal di-nitro compounds. In
the first step, chloro epoxide A1 is reacted with t-butylamine to
provide hydroxy heterocyclic compound B1. Mesylation of the
hydroxyl group of heterocyclic compound B1 with methylsulfonyl
chloride gives mesylate C1, which upon reacting with NaNO.sub.2
generates cyclic mono-nitro compound D1. Further nitration of
compound D1 can be carried out using NaNO.sub.2 in the presence of
NO.sub.2O.sub.8 and K.sub.3Fe(CN).sub.6 to provide geminal di-nitro
heterocyclic compound E1. Reacting compound E1 with boron
trifluoride etherate and acetyl bromide F provides the desired
product G1. Further description of related synthetic procedures are
described in, for example, Archibald et al. in J. Org. Chem. 1990,
55, 2920-2924; U.S. Pat. No. 7,507,842; and J. P. Agrawal, R. D.
Hodgson, Organic Chemistry of Explosives, Wiley & Sons,
England, 2007 and references cited therein.
[0135] This synthetic procedure illustrated in Scheme 1 and
described above is contemplated to be applicable to preparing
compounds having various substituents at the R.sub.1, R.sub.2,
R.sub.3 and R.sub.4 positions. If a particular epoxide compound
embraced by A1 should contain a functional group sensitive to one
or more of the synthetic transformations in Scheme 1, then standard
protecting group strategies are contemplated to be applied. For
further description of protecting group strategies and procedures,
see, for example, Greene, T. W.; Wuts, P. G. M. Protective Groups
in Organic Synthesis, 2.sup.nd ed.; Wiley, New York, 1991.
##STR00122##
[0136] Scheme 2 illustrates a more specific embodiment of the
synthetic route shown in Scheme 1 when m is 0. In the first step,
epoxide A2 is reacted with t-butylamine to provide hydroxyl
azetidine B2. Mesylation of the hydroxyl group of azetidine B2 with
methylsulfonyl chloride gives azetidine mesylate C2, which upon
reacting with NaNO.sub.2 generates mono-nitro azetidine D2. Further
nitration of mono-nitro azetidnine D2 with NaNO.sub.2 in the
presence of Na.sub.2S.sub.2O.sub.8 and K.sub.3Fe(CN).sub.6
furnishes the geminal di-nitro azetidine E2. Reaction of azetidine
E2 with boron trifluoride etherate and acetyl bromide compound F
provides the desired di-nitro azetidine product G2. This synthetic
procedure is contemplated to be applicable to preparing compounds
having various substituents at the R.sub.1, R.sub.2, R.sub.3 and
R.sub.4 positions. If a particular epoxide compound embraced by A2
should contain a functional group sensitive to one or more of the
synthetic transformations in Scheme 2, then standard protecting
group strategies are contemplated to be applied. For further
description of protecting group strategies and procedures, see, for
example, Greene, T. W.; Wuts, P. G. M. Protective Groups in Organic
Synthesis, 2.sup.nd ed.; Wiley, New York, 1991. Furthermore,
mono-nitro compounds can be prepared by treating mono-nitro
compound D2 with a Lewis Acid (e.g., boron trifluoride etherate)
and acetyl bromide compound F to provide the desired mono-nitro
product.
##STR00123##
[0137] Scheme 3 illustrates another more particular embodiment of
the synthetic route shown in Scheme 1 when both R.sub.1 and R.sub.2
are hydrogen and m is 0. In the first step, commercially available
epichlorohydrin A3 is reacted with t-butylamine to provide hydroxyl
azetidine B3. Mesylation of the hydroxyl group of azetidine B3 with
methylsulfonyl chloride gives azetidine mesylate C3, which upon
reacting with NaNO.sub.2 generates mono-nitro azetidine D3. Further
nitration of mono-nitro azetidine D3 with NaNO.sub.2 in the
presence of Na.sub.2S.sub.2O.sub.8 and K.sub.3Fe(CN).sub.6
furnishes the geminal di-nitro azetidine E3. Reaction of azetidine
E3 with boron trifluoride etherate and bromoacetyl bromide provides
the desired di-nitro azetidine F3. Further description of related
synthetic procedures are described in, for example, Archibald et
al. in J. Org. Chem. 1990, 55, 2920-2924; U.S. Pat. No. 7,507,842;
and J. P. Agrawal, R. D. Hodgson, Organic Chemistry of Explosives,
Wiley & Sons, England, 2007 and references cited therein.
Furthermore, mono-nitro compounds can be prepared by treating
mono-nitro compound D3 with a Lewis Acid (e.g., boron trifluoride
etherate) and acetyl bromide compound F to provide the desired
mono-nitro product.
##STR00124##
[0138] Scheme 4 illustrates an alternative exemplary procedure for
preparing cyclic geminal di-nitro compounds. In the first step,
heterocyclic compound A4 is reacted with an oxidant, such as
pyridinium dichromate (PDC), to provide heterocyclic ketone B4.
Reaction of ketone B4 with hydroxylamine gives heterocyclic oxime
C4, which upon reaction with N-bromosuccinimide (NBS) produces
bromo nitro compound D4. Reaction of compound D4 with NaBH.sub.4
furnishes mono-nitro compound E4. Reaction of mono-nitro compound
E4 with NaNO.sub.2 in the presence of Na.sub.2S.sub.2O.sub.8 and
K.sub.3Fe(CN).sub.6 provides geminal di-nitro heterocyclic compound
F4. Reaction of compound F4 with a deprotecting agent and acetyl
bromide compound F provides the desired cyclic geminal di-nitro
product G4. Further description of related synthetic procedures are
described in, for example, Archibald et al. in J. Org. Chem. 1990,
55, 2920-2924; U.S. Pat. No. 7,507,842; and J. P. Agrawal, R. D.
Hodgson, Organic Chemistry of Explosives, Wiley & Sons,
England, 2007 and references cited therein. Furthermore, mono-nitro
compounds can be prepared by treating mono-nitro compound D4 with a
deprotecting agent and acetyl bromide compound F to provide the
desired mono-nitro product.
##STR00125##
[0139] Scheme 5 illustrates yet another exemplary procedure for
preparing cyclic geminal di-nitro compounds with initial steps
different from those shown in Scheme 4. In the first step,
heterocyclic compound A4 is reacted with methylsulfonyl chloride to
provide heterocyclic mesylate B5. Reaction of mesylate B5 with
NaNO.sub.2 gives mono-nitro compound E4. Nitration of compound E4
with NaNO.sub.2 in the presence of Na.sub.2S.sub.2O.sub.8 and
K.sub.3Fe(CN).sub.6 provides geminal di-nitro compound F4. Reaction
of compound F4 with a deprotecting agent and acetyl bromide
compound F provides the desired di-nitro product G4. Further
description of related synthetic procedures are described in, for
example, Archibald et al. in J. Org. Chem. 1990, 55, 2920-2924;
U.S. Pat. No. 7,507,842; and J. P. Agrawal, R. D. Hodgson, Organic
Chemistry of Explosives, Wiley & Sons, England, 2007 and
references cited therein.
##STR00126##
[0140] The synthetic route illustrated in Scheme 6 depicts an
exemplary method for preparing cyclic vicinal di-nitro compounds.
In the first step, cycloalkene A6 is reacted with N.sub.2O.sub.4 to
provide vicinal di-nitro compound B6. Reaction of compound B6 with
a deprotecting agent and acetyl bromide compound F provides the
desired vicinal di-nitro product C6. Further description of related
synthetic procedures are described in, for example, Archibald et
al. in J. Org. Chem. 1990, 55, 2920-2924; U.S. Pat. No. 7,507,842;
and J. P. Agrawal, R. D. Hodgson, Organic Chemistry of Explosives,
Wiley & Sons, England, 2007 and references cited therein. This
synthetic procedure is contemplated to be applicable to preparing
compounds having various substituents at the R.sub.1, R.sub.2,
R.sub.3 and R.sub.4 positions. If a particular cycloalkene compound
embraced by A6 should contain a functional group sensitive to one
or more of the synthetic transformations in Scheme 6, then standard
protecting group strategies are contemplated to be applied. For
further description of protecting group strategies and procedures,
see, for example, Greene, T. W.; Wuts, P. G. M. Protective Groups
in Organic Synthesis, 2.sup.nd ed.; Wiley, New York, 1991.
##STR00127##
[0141] The synthetic route illustrated in Scheme 7 depicts a
general method for preparing cyclic mono-nitro compounds. In the
first step, chloro epoxide A7 is reacted with t-butylamine to
provide hydroxy heterocyclic compound B7. Mesylation of the
hydroxyl group of heterocyclic compound B7 with methylsulfonyl
chloride gives mesylate C7 which upon reacting with NaNO.sub.2
generates cyclic mono-nitro compound D7. Reaction of compound D7
with boron trifluoride etherate and acetyl bromide F provides the
desired product G7. Further description of related synthetic
procedures are described in, for example, Archibald et al. in J.
Org. Chem. 1990, 55, 2920-2924; U.S. Pat. No. 7,507,842; and J. P.
Agrawal, R. D. Hodgson, Organic Chemistry of Explosives, Wiley
& Sons, England, 2007 and references cited therein. This
synthetic procedure illustrated in Scheme 7 is contemplated to be
applicable to preparing compounds having various substituents at
the R.sub.1, R.sub.2, R.sub.3 and R.sub.4 positions. If a
particular epoxide compound embraced by A7 should contain a
functional group sensitive to one or more of the synthetic
transformations in Scheme 7, then standard protecting group
strategies are contemplated to be applied. For further description
of protecting group strategies and procedures, see, for example,
Greene, T. W.; Wuts, P. G. M. Protective Groups in Organic
Synthesis, 2.sup.nd ed.; Wiley, New York, 1991.
##STR00128##
[0142] The synthetic routes described above can be modified to
prepare compounds having an alkyl halide attached to the ring
nitrogen atom. Exemplary synthetic procedures for preparing such
compounds include reducing the amide group of compound G1-G4, G7,
and C6 to an amine. Alternatively, compound F used in the
procedures above could be replaced with an appropriately protected
alkylhalide, such that after the alkylation reaction, the protected
alkyl group attached to the ring nitrogen atom is deprotected and
converted to an alkyl chloride or bromide.
[0143] Scheme 8 depicts another exemplary method for preparing
cyclic mono-nitro and di-nitro compounds. Reaction of ketone B8
with hydroxylamine gives heterocyclic hydroxylamine C8, which upon
reaction with N-bromosuccinimide (NBS) produces bromo nitro
compound D8. Reaction of compound D8 with NaBH.sub.4 furnishes
mono-nitro compound E8. The hydroxyl protecting group (P, which may
be, for example, a tert-butyldimethylsilyl group) and the
1,2-dihydroxyethane protecting group are removed using standard
deprotection conditions. Exemplary deprotection conditions for
removing a tert-butyldimethyl silyl group include addition of
tetra-n-butylammonium fluoride. Exemplary deprotection conditions
for removing a 1,2-dihydroxyethane protecting group include
addition of hydrochloric acid and water. Hydroxy-ketone F8 can be
converted to .alpha.-bromo ketone G8 by first reacting compound F8
with methanesulfonyl chloride to form a mesylate and then adding
sodium bromide to form .alpha.-bromo ketone G8.
[0144] Di-nitro compounds can be prepared by reacting mono-nitro
compound E8 with NaNO.sub.2 in the presence of
Na.sub.2S.sub.2O.sub.8 and K.sub.3Fe(CN).sub.6 to provide geminal
di-nitro heterocyclic compound H8. The hydroxyl protecting group
(P, which may be, for example, a tert-butyldiimethyl silyl group)
and the 1,2-dihydroxyethane protecting group of compound H8 may be
removed using standard deprotection conditions. Exemplary
deprotection conditions for removing a tert-butyldiimethyl silyl
group include addition of tetra-n-butylammonium fluoride. Exemplary
deprotection conditions for removing a 1,2-dihydroxyethane
protecting group include addition of hydrochloric acid and water.
Hydroxy-ketone 18 can be converted to .alpha.-bromo ketone J8 by
first reacting compound 18 with methanesulfonyl chloride to form a
mesylate and then adding sodium bromide to form .alpha.-bromo
ketone J8. Further description of related synthetic procedures are
described in, for example, Archibald et al. in J. Org. Chem. 1990,
55, 2920-2924 and J. P. Agrawal, R. D. Hodgson, Organic Chemistry
of Explosives, Wiley & Sons, England, 2007 and references cited
therein.
##STR00129##
[0145] Scheme 9 illustrates an exemplary procedure for preparing
acyclic geminal di-nitro compounds. In the first step, protected
amino alcohol A9 is reacted with methylsulfonyl chloride to provide
mesylate B9. Reaction of mesylate B9 with NaNO.sub.2 gives
mono-nitro compound E9. Nitration of compound E9 with NaNO.sub.2 in
the presence of Na.sub.2S.sub.2O.sub.8 and K.sub.3Fe(CN).sub.6
provides geminal di-nitro compound F9. Reaction of compound F9 with
a deprotecting agent and acetyl bromide compound F provides the
desired di-nitro product G9. Further description of related
synthetic procedures are described in, for example, Archibald et
al. in J. Org. Chem. 1990, 55, 2920-2924; U.S. Pat. No. 7,507,842;
and J. P. Agrawal, R. D. Hodgson, Organic Chemistry of Explosives,
Wiley & Sons, England, 2007 and references cited therein.
##STR00130##
III. THERAPEUTIC APPLICATIONS OF COMBINATION THERAPY WITH INORGANIC
NITRITE SALT AND NITRITE-REDUCTASE PROMOTER
[0146] The invention provides methods for treating medical
disorders using an inorganic nitrite salt in combination with an
allosteric modulator of hemoglobin. The methods are contemplated to
provide particular advantages in treating or preventing various
medical disorders, such as a disorder selected from the group
consisting of cancer, a cardiovascular disorder, an ischemic
condition, a hemolytic condition, and a bacterial infection.
Various aspects of the therapeutic methods are described in detail
below.
[0147] A. General Therapeutic Methods
[0148] The therapeutic methods described herein are particularly
well-suited for treatment of diseases associated with hypoxic
conditions or ischemic conditions, or otherwise may be treated or
prevented using increased levels of nitric oxide. Accordingly, one
aspect of the invention provides a method of treating or preventing
a disorder selected from the group consisting of cancer, a
cardiovascular disorder, an ischemic condition, a hemolytic
condition, or a bacterial infection. The method comprises
administering to a patient in need thereof a therapeutically
effective amount of (i) an inorganic nitrite salt, and (ii) a
nitrite reductase promoter, which preferably is an allosteric
modulator of hemoglobin that promotes nitrite reductase
activity.
[0149] Exemplary Cancers
[0150] Exemplary types of cancer contemplated to be treated include
brain cancer, bladder cancer, breast cancer, cervical cancer, colon
cancer, colorectal cancer, endometrial cancer, esophageal cancer,
leukemia, lung cancer, liver cancer, melanoma, ovarian cancer,
pancreatic cancer, prostate cancer, rectal cancer, renal cancer,
stomach cancer, testicular cancer, or uterine cancer.
[0151] In certain embodiments, the cancer is a vascularized tumor,
solid tumor, squamous cell carcinoma, adenocarcinoma, small cell
carcinoma, melanoma, glioma, neuroblastoma, sarcoma (e.g., an
angiosarcoma or chondrosarcoma), larynx cancer, parotid cancer,
bilary tract cancer, thyroid cancer, acral lentiginous melanoma,
actinic keratoses, acute lymphocytic leukemia, acute myeloid
leukemia, adenoid cycstic carcinoma, adenomas, adenosarcoma,
adenosquamous carcinoma, anal canal cancer, anal cancer, anorectum
cancer, astrocytic tumor, bartholin gland carcinoma, basal cell
carcinoma, biliary cancer, bone cancer, bone marrow cancer,
bronchial cancer, bronchial gland carcinoma, carcinoid,
cholangiocarcinoma, chondosarcoma, choriod plexus
papilloma/carcinoma, chronic lymphocytic leukemia, chronic myeloid
leukemia, clear cell carcinoma, connective tissue cancer,
cystadenoma, digestive system cancer, duodenum cancer, endocrine
system cancer, endodermal sinus tumor, endometrial hyperplasia,
endometrial stromal sarcoma, endometrioid adenocarcinoma,
endothelial cell cancer, ependymal cancer, epithelial cell cancer,
Ewing's sarcoma, eye and orbit cancer, female genital cancer, focal
nodular hyperplasia, gallbladder cancer, gastric antrum cancer,
gastric fundus cancer, gastrinoma, glioblastoma, glucagonoma, heart
cancer, hemangiblastomas, hemangioendothelioma, hemangiomas,
hepatic adenoma, hepatic adenomatosis, hepatobiliary cancer,
hepatocellular carcinoma, Hodgkin's disease, ileum cancer,
insulinoma, intaepithelial neoplasia, interepithelial squamous cell
neoplasia, intrahepatic bile duct cancer, invasive squamous cell
carcinoma, jejunum cancer, joint cancer, Kaposi's sarcoma, pelvic
cancer, large cell carcinoma, large intestine cancer,
leiomyosarcoma, lentigo maligna melanomas, lymphoma, male genital
cancer, malignant melanoma, malignant mesothelial tumors,
medulloblastoma, medulloepithelioma, meningeal cancer, mesothelial
cancer, metastatic carcinoma, mouth cancer, mucoepidermoid
carcinoma, multiple myeloma, muscle cancer, nasal tract cancer,
nervous system cancer, neuroepithelial adenocarcinoma nodular
melanoma, non-epithelial skin cancer, non-Hodgkin's lymphoma, oat
cell carcinoma, oligodendroglial cancer, oral cavity cancer,
osteosarcoma, papillary serous adenocarcinoma, penile cancer,
pharynx cancer, pituitary tumors, plasmacytoma, pseudosarcoma,
pulmonary blastoma, rectal cancer, renal cell carcinoma,
respiratory system cancer, retinoblastoma, rhabdomyosarcoma,
sarcoma, serous carcinoma, sinus cancer, skin cancer, small cell
carcinoma, small intestine cancer, smooth muscle cancer, soft
tissue cancer, somatostatin-secreting tumor, spine cancer, squamous
cell carcinoma, striated muscle cancer, submesothelial cancer,
superficial spreading melanoma, T cell leukemia, tongue cancer,
undifferentiated carcinoma, ureter cancer, urethra cancer, urinary
bladder cancer, urinary system cancer, uterine cervix cancer,
uterine corpus cancer, uveal melanoma, vaginal cancer, verrucous
carcinoma, VIPoma, vulva cancer, well differentiated carcinoma, or
Wilms tumor.
[0152] The therapeutic methods may optionally comprise exposing the
patient to a chemotherapeutic agent or radiation. One exemplary
form of radiation is gamma rays, such as those produced from a
.sup.137Cs source. The amount of radiation can be optimized for
particular conditions. In certain embodiments, the quantity of
radiation applied to the patient is at least about 2 Gy, about 5
Gy, about 10 Gy, or about 15 Gy. Exemplary chemotherapeutic agents
include azacitidine, azathioprine, bleomycin, carboplatin,
capecitabine, carmustine, cisplatin, chlorambucil,
cyclophosphamide, cytarabine, dacarbazine, daunorubicin, docetaxel,
doxifluridine, doxorubicin, epirubicin, epothilone, etoposide,
fluorouracil, fulvestrant, gemcitabine, hydroxyurea, idarubicin,
imatinib, lomustine, mechlorethamine, mercaptopurine, methotrexate,
mitoxantrone, oxaliplatin, paclitaxel, pemetrexed, procarbazine,
raloxifene, teniposide, temozolomide, thiotepa, tioguanine,
tamoxifen, toremifene, valrubicin, vinblastine, vincristine,
vindesine, vinorelbine, and pharmaceutically acceptable salts
thereof
Exemplary Cardiovascular Disorders
[0153] Exemplary cardiovascular disorders include pulmonary
hypertension, systemic hypertension, angina (e.g., Prinzmetal's
angina), Cardiac syndrome X, myocardial infarction, peripheral
artery disease, Raynaud's disease, pulmonary embolism, and
intravascular thrombosis. In certain embodiments, the
cardiovascular disorder is pulmonary hypertension, systemic
hypertension, angina (e.g., Prinzmetal's angina), Cardiac syndrome
X, myocardial infarction, peripheral artery disease, or Raynaud's
disease.
Exemplary Ischemic Conditions
[0154] Exemplary ischemic conditions include stroke, an ischemic
central nervous system event, cardiac ischemia syndrome, myocardial
ischemia, and tissue damage due to hypoxia.
Exemplary Hemolytic Conditions
[0155] Exemplary hemolytic conditions include sickle cell disease
(including sickle cell crisis), thalassemia, hemoglobin C disease,
hemoglobin SC disease, sickle thalassemia, hereditary
spherocytosis, hereditary elliptocytosis, hereditary ovalcytosis,
glucose-6-phosphate deficiency and other red blood cell enzyme
deficiencies, paroxysmal nocturnal hemoglobinuria (PNH), paroxysmal
cold hemoglobinuria (PCH), thrombotic thrombocytopenic
purpura/hemolytic uremic syndrome (TTP/HUS), idiopathic autoimmune
hemolytic anemia, drug-induced immune hemolytic anemia, secondary
immune hemolytic anemia, non-immune hemolytic anemia caused by
chemical or physical agents, malaria, falciparum malaria,
bartonellosis, babesiosis, clostridial infection, severe
haemophilus influenzae type b infection, extensive burns,
transfusion reaction, rhabdomyolysis (myoglobinemia), transfusion
of aged blood, cardiopulomonary bypass, and hemodialysis.
Exemplary Bacterial Infections
[0156] The bacterial infection may be a gram-positive bacterial
infection or a gram-negative bacterial infection. In certain
embodiments, the bacterial infection is a gram-positive cocci
bacterial infection or a gram-positive bacilli bacterial infection.
In certain other embodiments, the bacterial infection is a
gram-negative bacterial infection. In certain other embodiments,
the bacterial infection is a gram-negative cocci bacterial
infection or a gram-negative bacilli bacterial infection.
[0157] The type of bacterial infection can also be characterized
according to whether the bacterial infection is caused by anaerobic
or aerobic bacteria. In certain embodiments, the bacterial
infection is an anaerobic bacterial infection. In certain other
embodiments, the bacterial infection is an aerobic bacterial
infection.
[0158] In certain embodiments, the bacterial infection is a
mycobacterial infection. In more particular embodiments, the
bacterial infection is an infection of bacteria selected from the
group consisting of Mycobacterium tuberculosis, Staphylococcus
aureus, Staphylococcus epidermidis, Enterococcus faecalis,
Enterococcus faecium, Streptococcus pneumoniae, Streptococcus
pyogenes, Mycobacterium smegmatis, Bacillus anthracis, Escherichia
coli, Proteus mirabilis, Pseudomonas aeruginosa, Acinetobacter
baumannii, Yersinia enterocolytica, Francisella tularensis,
Eubacterium lentum, Bacteroides fragilis, Fusobacterium nucleatum,
Porphyromonas asaccharolyticus, Clostridium perfringens, and
Clostridium difficile. In still other embodiments the bacterial
infection is an infection of Mycobacterium tuberculosis bacteria
(abbreviated as "MTB" or "TB").
[0159] In certain other embodiments, the bacterial infection is due
to a member of the genus Peptostreptococci, a Peptostreptococci
asaccharolyticus, a Peptostreptococci magnus, a Peptostreptococci
micros, a Peptostreptococci prevotii, a member of the genus
Porphyromonas, a Porphyromonas asaccharolytica, a Porphyromonas
canoris, a Porphyromonas gingivalis, a Porphyromonas macaccae, a
member of the genus Actinomyces, an Actinomyces israelii, an
Actinomyces odontolyticus, a member of the genus Clostridium, a
Clostridium innocuum, a Clostridium clostridioforme, a Clostridium
difficile, a member of the genus Anaerobiospirillum, a member of
the genus Bacteroides, a Bacteroides tectum, a Bacteroides
ureolyticus, a Bacteroides gracilis (Campylobacter gracilis), a
member of the genus a Prevotella, a Prevotella intermedia, a
Prevotella heparinolytica, a Prevotella orisbuccae, a Prevotella
bivia, a Prevotella melaninogenica, a member of the genus
Fusobacterium, a Fusobacterium naviforme, a Fusobacterium
necrophorum, a Fusobacterium varium, a Fusobacterium ulcerans, a
Fusobacterium russii, a member of the genus Bilophila, or a
Bilophila wadsworthia.
[0160] In certain other embodiments, the bacterial infection is due
to an antibiotic-resistant bacteria, both aerobic and anaerobic,
Gram positive and Gram negative.
Additional Medical Conditions
[0161] Additional medical conditions contemplated for treatment or
prevention using compositions described herein include nitrogen
oxide related rheumatoid arthritis, diabetes (including
neuropathies and vasculopathies), and systemic lupus
erythematosus.
Additional Considerations
[0162] The patient is preferably a human, such as a human suffering
from a tumor. The particular combination of inorganic nitrite salt
and allosteric modulator of hemoglobin may be selected according to
the medical disorder suffered by the patient. For example, in
certain embodiments, the inorganic nitrite salt is one of the
generic or specific nitrite salts described in Section II, such as
alkali metal nitrite, in particular, sodium nitrite. In certain
other embodiments, the allosteric modulator of hemoglobin is one of
the generic or specific allosteric modulators of hemoglobin
described in Section II, such as a compound of Formula I, a
compound embraced by one of the further embodiments describing
definitions for certain variables of Formula I, a compound of
Formula I-A, or a compound embraced by one of the further
embodiments describing definitions for certain variables of Formula
I-A.
[0163] Further yet, for example, with regards to Formula I, in
certain embodiments, the compound corresponds to Formula I where
A.sup.1 is --C(O)--. In certain other embodiments, A.sup.1 is
--(C(R.sup.3).sub.2).sub.xC(O)(C(R.sup.3).sub.2).sub.x--. In
certain other embodiments, A.sup.1 is
--C(O)(C(R.sup.3).sub.2).sub.x--.
[0164] In certain embodiments, A.sup.2 is N. In certain other
embodiments, A.sup.2 is --C(R.sup.4)--.
[0165] In certain embodiments, R.sup.1 is halogen,
--OS(O).sub.2R.sup.5, or --OC(O)CF.sub.3. In certain other
embodiments, R.sup.1 is halogen. In certain other embodiments,
R.sup.1 is --OS(O).sub.2R.sup.5. In certain other embodiments,
R.sup.1 is --OC(O)CF.sub.3. In certain other embodiments, R.sup.1
is chloro, bromo, --OS(O).sub.2-(para-methylphenyl),
--OS(O).sub.2CH.sub.3, --OS(O).sub.2CF.sub.3, or --OC(O)CF.sub.3.
In certain embodiments, R.sup.1 is bromo.
[0166] In certain embodiments, m is 2. In certain other
embodiments, m is 1. In certain embodiments, n is 0. In certain
other embodiments, n is 1. In certain other embodiments, n is 2. In
certain embodiments, p is 1. In certain other embodiments, p is 2.
In certain other embodiments, p is 3.
[0167] In certain embodiments, the allosteric modulator of
hemoglobin is a compound of Formula I-A:
##STR00131##
[0168] or a pharmaceutically acceptable salt or solvate thereof,
wherein:
[0169] A is N or C(H);
[0170] R.sup.1 is chloro, bromo,
--OS(O).sub.2--(C.sub.1-C.sub.6alkyl),
--OS(O).sub.2--(C.sub.1-C.sub.6haloalkyl),
--OS(O).sub.2-(para-methylphenyl), or --OC(O)CF.sub.3;
[0171] R.sup.2 represents independently for each occurrence
hydrogen or methyl;
[0172] y represents independently for each occurrence 1 or 2.
[0173] In certain embodiments, A is N. In certain other
embodiments, A is C(H).
[0174] In certain embodiments, R.sup.1 is chloro. In certain other
embodiments, R.sup.1 is bromo.
[0175] In certain embodiments, R.sup.1 is
--OS(O).sub.2--(C.sub.1-C.sub.6alkyl),
--OS(O).sub.2--(C.sub.1-C.sub.6haloalkyl), or
--OS(O).sub.2-(para-methylphenyl). In certain other embodiments,
R.sup.1 is --OS(O).sub.2CH.sub.3, --OS(O).sub.2CF.sub.3, or
--OS(O).sub.2-(para-methylphenyl). In certain other embodiments,
R.sup.1 is --OC(O)CF.sub.3.
[0176] In certain embodiments, R.sup.2 is hydrogen or methyl. In
certain embodiments, R.sup.2 is hydrogen.
[0177] In certain embodiments, y is 1. In certain embodiments, one
occurrence of y is 1, and the other occurrence of y is 2. In
certain other embodiments, y is 2.
[0178] In certain embodiments, the allosteric modulator of
hemoglobin is
##STR00132##
or a pharmaceutically acceptable salt or solvate thereof. In
certain other embodiments, the allosteric modulator of hemoglobin
is
##STR00133##
[0179] In certain other embodiments, the allosteric modulator of
hemoglobin is one of the compounds listed in Tables 1 and 2 herein
or a pharmaceutically acceptable salt or solvate thereof
[0180] The description above describes multiple embodiments
relating to methods of treating various disorders using an
inorganic nitrite salt in combination with an allosteric modulator
of hemoglobin. The patent application specifically contemplates all
combinations of the embodiments. For example, the invention
contemplates treating a tumor by administering a therapeutically
effective amount of sodium nitrite in combination with a compound
of Formula I-A wherein A is N, R.sup.1 is chloro or bromo, and
R.sup.2 is hydrogen. Further, for example, the invention
contemplates treating a tumor by administering a therapeutically
effective amount of sodium nitrite in combination with a compound
of Formula II wherein A.sup.1 is --C(O)--, A.sup.2 is N(R.sup.5),
and R.sup.2 and R.sup.3 are hydrogen.
[0181] B. Methods of Increasing the Amount of Nitric Oxide Produced
by Hemoglobin
[0182] Another aspect of the invention provides a method of
increasing the amount of nitric oxide produced by hemoglobin in a
patient. The method comprises administering to a patient in need
thereof a therapeutically effective amount of (i) an inorganic
nitrite salt, and (ii) a nitrite reductase promoter, preferably an
allosteric modulator of hemoglobin that promotes nitrite reductase
activity. In certain embodiments, the allosteric modulator of
hemoglobin is administered at a dosage sufficient to cause a ten
percent increase in the rate at which hemoglobin converts nitrite
to nitric oxide in vivo. In certain other embodiments, the dose of
inorganic nitrite salt and dose of allosteric modulator of
hemoglobin are sufficient to cause a ten percent increase in the
rate at which hemoglobin converts nitrite to nitric oxide in
vivo.
[0183] C. Methods of Preventing Sickling of a Red Blood Cell
[0184] Another aspect of the invention provides a method of
preventing sickling of a red blood cell susceptible to sickling.
The method comprises exposing said red blood cell to an effective
amount of (i) an inorganic nitrite salt, and (ii) a nitrite
reductase promoter (which preferably is an allosteric modulator of
hemoglobin that promotes nitrite reductase activity) to prevent
sickling of the red blood cell.
[0185] In certain embodiments, the red blood cell is a red blood
cell in a patient suffering from sickle cell anemia. In certain
embodiments, less than 10% of a population of said red blood cells
convert to sickle form when exposed to an effective amount of (i)
an inorganic nitrite salt, and (ii) an allosteric modulator of
hemoglobin that promotes nitrite reductase activity, under hypoxic
conditions. In certain embodiments, the hypoxic condition is
characterized by a pO.sub.2 of less than about 10 mm Hg.
[0186] D. Dosing Amounts
[0187] Generally, the combination of pharmaceutical agents is
delivered to the patient in an effective amount. Actual dosage
levels of the active ingredients in the pharmaceutical compositions
of this invention may be varied so as to obtain an amount of the
active ingredient which is effective to achieve the desired
therapeutic response for a particular patient, composition, and
mode of administration, without being toxic to the patient. The
selected dosage level will depend upon a variety of factors
including the activity of the particular compound of the present
invention employed, the route of administration, the time of
administration, the rate of excretion or metabolism of the
particular compound being employed, the rate and extent of
absorption, the duration of the treatment, other drugs, compounds
and/or materials used in combination with the particular compound
employed, the age, sex, weight, condition, general health and prior
medical history of the patient being treated, and like factors well
known in the medical arts.
[0188] In general, a suitable daily dose of a compound of the
invention will be that amount of the compound which is the lowest
dose effective to produce a therapeutic effect. If desired, the
effective daily dose of the active compound may be administered as
two, three, four, five, six or more sub-doses administered
separately at appropriate intervals throughout the day, optionally,
in unit dosage forms. Preferred dosing is one administration per
day.
[0189] In certain embodiments, an inorganic nitrite is administered
at a daily dosage of from about 0.1 .mu.g/kg to about 10 mg/kg,
about 1 .mu.g to about 5 mg/kg, about 0.05 mg/kg to about 10 mg/kg,
about 0.1 mg/kg to about 5 mg/kg, about 0.5 mg/kg to about 5 mg/kg,
about 0.5 mg/kg to about 3 mg/kg, about 0.1 mg/kg to about 1.5
mg/kg, about 0.1 mg/kg to about 0.35 mg/kg, about 0.35 mg/kg to
about 0.75 mg/kg, or about 0.75 mg/kg to about 1 mg/kg. In certain
other embodiments, an inorganic nitrite may be administered in an
amount such that the plasma concentration of nitrite ion is from
about 0.05 .mu.M to about 200 .mu.M, about 0.1 .mu.M to about 100
.mu.M, about 0.5 .mu.M to about 100 .mu.M, about 0.1 .mu.M to about
100 .mu.M, or about 1 .mu.M to about 100 .mu.M for a desired period
of time. In certain embodiments, the desired plasma concentration
is maintained for a period of from about 1 hour to about 20 hours,
about 1 hour to about 10 hours, or about 1 hour to about 5
hours.
[0190] C. Combination Therapy
[0191] The therapeutic methods embrace combination therapy, which
includes the administration of an inorganic nitrite salt in
combination with an allosteric modulator of hemoglobin as part of a
specific treatment regimen intended to provide the beneficial
effect from the co-action of these therapeutic agents.
Administration of these therapeutic agents in combination typically
is carried out over a defined time period (e.g., hours or days
depending upon the combination selected). The combination therapy
may involve administration of two or more of these therapeutic
agents as part of separate monotherapy regimens that result in the
combinations of the present invention. Combination therapy also
includes administration of these therapeutic agents in a sequential
manner, that is, wherein each therapeutic agent is administered at
a different time, as well as administration of these therapeutic
agents, or at least two of the therapeutic agents, in a
substantially simultaneous manner. Substantially simultaneous
administration can be accomplished, for example, by administering
to the subject a single capsule having a fixed ratio of each
therapeutic agent or in multiple, single capsules for each of the
therapeutic agents. Sequential or substantially simultaneous
administration of each therapeutic agent can be effected by any
appropriate route including, but not limited to, oral routes,
intravenous routes, intramuscular routes, and direct absorption
through mucous membrane tissues.
[0192] It is understood that the therapeutic agents can be
administered by the same route or by different routes. For example,
a first therapeutic agent of the combination selected may be
administered by intravenous administration while the other
therapeutic agent(s) of the combination may be administered orally.
Alternatively, for example, all therapeutic agents may be
administered orally or all therapeutic agents may be administered
by intravenous injection.
IV. TREATING PATIENTS WITH REDUCED BLOOD VOLUME AND/OR IN NEED OF
TRANSFUSION
[0193] One aspect of the invention provides a method of treating a
patient suffering from reduced blood volume. The method comprises
administering to a patient in need thereof a blood product by
injection and a therapeutic agent selected from the group
consisting of an organonitro compound of Formula I, organonitro
compound of Formula II, hemoglobin conjugate of Formula III,
hemoglobin conjugate of Formula IV, and an erythrocyte cell that
has been exposed to an organonitro compound of Formula I or II;
wherein Formula I is represented by:
##STR00134##
[0194] or a pharmaceutically acceptable salt or solvate thereof,
wherein: [0195] A.sup.1 is --C(O)-- or
--(C(R.sup.3).sub.2).sub.xC(O)(C(R.sup.3).sub.2).sub.x--; [0196]
A.sup.2 is N or --C(R.sup.4)--; [0197] R.sup.1 is halogen,
--OS(O).sub.2R.sup.5, or --OC(O)CF.sub.3; [0198] R.sup.2 is
C.sub.1-C.sub.6alkyl; [0199] R.sup.3 and R.sup.4 each represent
independently for each occurrence hydrogen or C.sub.1-C.sub.5alkyl;
[0200] R.sup.5 is C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6haloalkyl,
aryl, or aralkyl; [0201] m and p are independently 1, 2, or 3; and
[0202] n and x each represent independently for each occurrence 0,
1, 2, or 3;
[0203] Formula II is represented by:
##STR00135##
[0204] or a pharmaceutically acceptable salt or solvate thereof:
wherein: [0205] A.sup.1 is --C(O)-- or
--(C(R.sup.5).sub.2).sub.xC(O)(C(R.sup.5).sub.2).sub.x--; [0206]
A.sup.2 is --N(R.sup.5)-- or --C(R.sup.2)(R.sup.3)--; [0207]
R.sup.1 is halogen, --OS(O).sub.2R.sup.6, or --OC(O)CF.sub.3;
[0208] R.sup.2 and R.sup.3 each represent independently for each
occurrence hydrogen or C.sub.1-C.sub.6alkyl; or [0209] R.sup.2 and
R.sup.3 are taken together with the carbon atom to which they are
attached to form a 3-6 membered, saturated carbocyclic ring; [0210]
R.sup.4 is hydrogen or C.sub.1-C.sub.6alkyl; [0211] R.sup.5
represents independently for each occurrence hydrogen or
C.sub.1-C.sub.6alkyl; [0212] R.sup.6 is C.sub.1-C.sub.6alkyl,
C.sub.1-C.sub.6haloalkyl, aryl, or aralkyl; [0213] t is an integer
in the range from 1 to 12; and [0214] x represents independently
for each occurrence 0, 1, 2, or 3;
[0215] Formula III is represented by:
##STR00136##
[0216] or a pharmaceutically acceptable salt or solvate thereof,
wherein: [0217] A.sup.1 is --C(O)-- or
--C(O)(C(R.sup.3).sub.2).sub.x--; [0218] A.sup.2 is N or
--C(R.sup.4)--; [0219] R.sup.2 is C.sub.1-C.sub.6alkyl; [0220]
R.sup.3 and R.sup.4 each represent independently for each
occurrence hydrogen or C.sub.1-C.sub.5alkyl; [0221] m and p are
independently 1, 2, or 3; [0222] n is 0, 1, 2, or 3; [0223] x is 1,
2, or 3; and [0224] z is an integer from 1 to 10; and
[0225] Formula IV is represented by:
##STR00137##
[0226] or a pharmaceutically acceptable salt or solvate thereof,
wherein: [0227] A.sup.1 is --C(O)-- or
--C(O)(C(R.sup.5).sub.2).sub.x--; [0228] A.sup.2 is --N(R.sup.5)--
or --C(R.sup.2)(R.sup.3)--; [0229] R.sup.2 and R.sup.3 each
represent independently for each occurrence hydrogen or
C.sub.1-C.sub.6alkyl; or [0230] R.sup.2 and R.sup.3 are taken
together with the carbon atom to which they are attached to form a
3-6 membered, saturated carbocyclic ring; [0231] R.sup.4 is
hydrogen or C.sub.1-C.sub.6alkyl; [0232] R.sup.5 represents
independently for each occurrence hydrogen or C.sub.1-C.sub.6alkyl;
[0233] t is an integer in the range from 1 to 12; [0234] x is 1, 2,
or 3; and [0235] z is an integer from 1 to 10.
[0236] In certain embodiments, the patient suffering from reduced
blood volume is suffering from hemorrhagic shock. Hemorrhagic shock
is characterized by rapid and significant loss of blood
(hypovolemia), resulting in the inadequate delivery of oxygen and
nutrients to meet metabolic demands. Compensatory mechanisms are
often activated to preserve perfusion selectively to the brain and
heart at the expense of other organ systems with progressive
development of shock at the cellular and tissue level due to blood
flow redistribution. The present method provides a treatment for
such hemorrhagic shock.
[0237] Another aspect of the invention provides a method of
performing a blood transfusion to a patient. The method comprises
administering to a patient in need thereof a blood product by
injection and a therapeutic agent selected from the group
consisting of an organonitro compound of Formula I, organonitro
compound of Formula II, hemoglobin conjugate of Formula III,
hemoglobin conjugate of Formula IV, and an erythrocyte cell that
has been exposed to an organonitro compound of Formula I or II,
wherein Formula I is represented by:
##STR00138##
[0238] or a pharmaceutically acceptable salt or solvate thereof,
wherein: [0239] A.sup.1 is --C(O)-- or
--(C(R.sup.3).sub.2).sub.xC(O)(C(R.sup.3).sub.2).sub.x--; [0240]
A.sup.2 is N or --C(R.sup.4)--; [0241] R.sup.1 is halogen,
--OS(O).sub.2R.sup.5, or --OC(O)CF.sub.3; [0242] R.sup.2 is
C.sub.1-C.sub.6alkyl; [0243] R.sup.3 and R.sup.4 each represent
independently for each occurrence hydrogen or C.sub.1-C.sub.5alkyl;
[0244] R.sup.5 is C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6haloalkyl,
aryl, or aralkyl; [0245] m and p are independently 1, 2, or 3; and
[0246] n and x each represent independently for each occurrence 0,
1, 2, or 3;
[0247] Formula II is represented by:
##STR00139##
[0248] or a pharmaceutically acceptable salt or solvate thereof:
wherein: [0249] A.sup.1 is --C(O)-- or
--(C(R.sup.5).sub.2).sub.xC(O)(C(R.sup.5).sub.2).sub.x--; [0250]
A.sup.2 is --N(R.sup.5)-- or --C(R.sup.2)(R.sup.3)--; [0251]
R.sup.1 is halogen, --OS(O).sub.2R.sup.6, or --OC(O)CF.sub.3;
[0252] R.sup.2 and R.sup.3 each represent independently for each
occurrence hydrogen or C.sub.1-C.sub.6alkyl; or [0253] R.sup.2 and
R.sup.3 are taken together with the carbon atom to which they are
attached to form a 3-6 membered, saturated carbocyclic ring; [0254]
R.sup.4 is hydrogen or C.sub.1-C.sub.6alkyl; [0255] R.sup.5
represents independently for each occurrence hydrogen or
C.sub.1-C.sub.6alkyl; [0256] R.sup.6 is C.sub.1-C.sub.6alkyl,
C.sub.1-C.sub.6haloalkyl, aryl, or aralkyl; [0257] t is an integer
in the range from 1 to 12; and [0258] x represents independently
for each occurrence 0, 1, 2, or 3;
[0259] Formula III is represented by:
##STR00140##
[0260] or a pharmaceutically acceptable salt or solvate thereof,
wherein: [0261] A.sup.1 is --C(O)-- or
--C(O)(C(R.sup.3).sub.2).sub.x--; [0262] A.sup.2 is N or
--C(R.sup.4)--; [0263] R.sup.2 is C.sub.1-C.sub.6alkyl; [0264]
R.sup.3 and R.sup.4 each represent independently for each
occurrence hydrogen or C.sub.1-C.sub.5alkyl; [0265] m and p are
independently 1, 2, or 3; [0266] n is 0, 1, 2, or 3; [0267] x is 1,
2, or 3; and [0268] z is an integer from 1 to 10; and
[0269] Formula IV is represented by:
##STR00141##
[0270] or a pharmaceutically acceptable salt or solvate thereof,
wherein: [0271] A.sup.1 is --C(O)-- or
--C(O)(C(R.sup.5).sub.2).sub.x--; [0272] A.sup.2 is --N(R.sup.5)--
or --C(R.sup.2)(R.sup.3)--; [0273] R.sup.2 and R.sup.3 each
represent independently for each occurrence hydrogen or
C.sub.1-C.sub.6alkyl; or [0274] R.sup.2 and R.sup.3 are taken
together with the carbon atom to which they are attached to form a
3-6 membered, saturated carbocyclic ring; [0275] R.sup.4 is
hydrogen or C.sub.1-C.sub.6alkyl; [0276] R.sup.5 represents
independently for each occurrence hydrogen or C.sub.1-C.sub.6alkyl;
[0277] t is an integer in the range from 1 to 12; [0278] x is 1, 2,
or 3; and [0279] z is an integer from 1 to 10.
[0280] In certain embodiments, the blood product comprises
erythrocyte cells. In certain embodiments, the blood product
comprises blood plasma. In certain other embodiments, the blood
product comprises erythrocyte cells and blood plasma.
[0281] In certain other embodiments, the blood product and
organonitro compound are administered to the patient
concurrently.
[0282] In certain embodiments, the blood product is administered to
the patient separately from the therapeutic agent.
[0283] In certain embodiments, the patient receives, by intravenous
injection, a single composition comprising blood product and the
therapeutic agent. In other certain embodiments, the patient
receives, by intravenous injection, a single composition comprising
a therapeutic agent, plasma, and erythrocyte cells. One exemplary
composition is provided below in Table 1.
TABLE-US-00004 TABLE 1 Exemplary Composition for Intravenous
Injection* Component Amount Erythrocyte cells (vol %) 35-60 Plasma
(mL) 17 Anticoagulant As needed (e.g., 4 mL) Therapeutic Agent
(e.g., As needed, such as, an ABDNAZ) amount to treat hemorrhagic
shock. *Amounts are based on a composition having a total volume of
282 mL.
[0284] In certain embodiments, the method further comprises
administering an alkali metal nitrite to the patient. In other
embodiments, the method further comprises administering sodium
nitrite to the patient.
[0285] In certain embodiments, the patient receives a single
composition comprising blood product, therapeutic agent, and an
alkali metal nitrite.
[0286] In certain embodiments, the therapeutic agent is an
organonitro compound of Formula I. In certain other embodiments,
the therapeutic agent is an erythrocyte cell that has been exposed
to an organonitro compound of Formula I, and said therapeutic agent
is administered by injection (such as intravenous injection).
[0287] In certain embodiments, A.sup.1 is --C(O)--, and A.sup.2 is
N.
[0288] In certain embodiments, R.sup.1 is bromo.
[0289] In certain embodiments, n is 0, and m is 2.
[0290] The description above describes multiple embodiments
relating to compounds of Formula I. The patent application
specifically contemplates all combinations of the embodiments. For
example, the invention contemplates a compound of Formula I wherein
A.sup.1 is --C(O)--, A.sup.2 is N, R.sup.1 is halogen, and n is
0.
[0291] In certain embodiments, the therapeutic agent is a compound
of Formula I-A:
##STR00142##
[0292] or a pharmaceutically acceptable salt or solvate thereof,
wherein: [0293] A is N or C(H); [0294] R.sup.1 is chloro, bromo,
--OS(O).sub.2--(C.sub.1-C.sub.6alkyl),
--OS(O).sub.2--(C.sub.1-C.sub.6haloalkyl),
--OS(O).sub.2-(para-methylphenyl), or --OC(O)CF.sub.3; [0295]
R.sup.2 represents independently for each occurrence hydrogen or
methyl; and [0296] y represents independently for each occurrence 1
or 2.
[0297] In certain embodiments, the therapeutic agent is compound
embraced by Formula I-A as defined by particular definitions for
variables in Formula I-A, such as where A is N. In certain other
embodiments, A is C(H).
[0298] In certain embodiments, R.sup.1 is chloro or bromo. In
certain embodiments, R.sup.1 is chloro. In certain other
embodiments, R.sup.1 is bromo. In certain embodiments, R.sup.1 is
--OS(O).sub.2--(C.sub.1-C.sub.6alkyl),
--OS(O).sub.2--(C.sub.1-C.sub.6haloalkyl), or
--OS(O).sub.2-(para-methylphenyl). In certain other embodiments,
R.sup.1 is --OS(O).sub.2CH.sub.3, --OS(O).sub.2CF.sub.3, or
--OS(O).sub.2-(para-methylphenyl). In certain other embodiments,
R.sup.1 is --OC(O)CF.sub.3.
[0299] In certain embodiments, R.sup.2 is hydrogen or methyl. In
certain embodiments, R2 is hydrogen.
[0300] In certain embodiments, y is 1. In certain embodiments, one
occurrence of y is 1, and the other occurrence of y is 2. In
certain other embodiments, y is 2.
[0301] The description above describes multiple embodiments
relating to compounds of Formula I-A. The patent application
specifically contemplates all combinations of the embodiments. For
example, the invention contemplates a compound of Formula I-A
wherein A is N, R.sup.1 is chloro or bromo, and R.sup.2 is
hydrogen.
[0302] In certain embodiments, the therapeutic agent is
##STR00143##
or a pharmaceutically acceptable salt thereof
[0303] In certain embodiments, the therapeutic agent an organonitro
compound embraced by Formula II as defined by particular
definitions for variables in Formula II, such as where A.sup.1 is
--C(O)--. In certain other embodiments, A.sup.1 is
--(C(R.sup.5).sub.2).sub.xC(O)(C(R.sup.5).sub.2).sub.x--. In
certain other embodiments, A.sup.1 is
--C(O)(C(R.sup.5).sub.2).sub.x--.
[0304] In certain embodiments, A.sup.2 is --N(R.sup.5)--. In
certain other embodiments, A.sup.2 is --C(R.sup.2)(R.sup.3)--.
[0305] In certain embodiments, R.sup.1 is halogen. In certain other
embodiments, R.sup.1 is --OS(O).sub.2R.sup.6. In certain other
embodiments, R.sup.1 is --OC(O)CF.sub.3. In certain other
embodiments, R.sup.1 is chloro, bromo,
--OS(O).sub.2-(para-methylphenyl), --OS(O).sub.2CH.sub.3,
--OS(O).sub.2CF.sub.3, or --OC(O)CF.sub.3. In certain embodiments,
R.sup.1 is bromo.
[0306] In certain embodiments, R.sup.2 and R.sup.3 each represent
independently for each occurrence hydrogen or C.sub.1-C.sub.6alkyl.
In certain other embodiments, R.sup.2 and R.sup.3 each represent
independently for each occurrence hydrogen, methyl, ethyl, or
propyl. In certain other embodiments, R.sup.2 and R.sup.3 each
represent independently for each occurrence hydrogen or methyl. In
certain embodiments, R.sup.2 and R.sup.3 are hydrogen.
[0307] In certain embodiments, R.sup.4 is hydrogen, methyl, ethyl,
propyl, butyl, or pentyl. In certain other embodiments, R.sup.4 is
methyl, ethyl or propyl. In certain other embodiments, R.sup.4 is
methyl.
[0308] In certain embodiments, R.sup.5 is hydrogen or methyl. In
certain other embodiments, R.sup.5 is hydrogen.
[0309] In certain embodiments, R.sup.6 is C.sub.1-C.sub.6alkyl or
C.sub.1-C.sub.6haloalkyl. In certain other embodiments, R.sup.6 is
methyl, ethyl, or trifluoromethyl. In certain other embodiments,
R.sup.6 is aryl, such as phenyl.
[0310] In certain embodiments, t is 1, 2, 3, 4, 5 or 6. In certain
other embodiments, t is 1, 2, or 3. In certain other embodiments, t
is 1. In certain embodiments, x is 1 or 2.
[0311] The description above describes multiple embodiments
relating to compounds of Formula II. The patent application
specifically contemplates all combinations of the embodiments. For
example, the invention contemplates a compound of Formula II
wherein A.sup.1 is --C(O)--, A.sup.2 is --N(R.sup.5)--, and R.sup.2
and R.sup.3 are hydrogen.
[0312] In certain embodiments, the therapeutic agent is a compound
of Formula II-A:
##STR00144##
[0313] or a pharmaceutically acceptable salt or solvate thereof:
wherein: [0314] A is --N(R.sup.5)-- or --C(R.sup.2)(R.sup.3)--;
[0315] R.sup.1 is chloro, bromo,
--OS(O).sub.2--(C.sub.1-C.sub.6alkyl),
--OS(O).sub.2--(C.sub.1-C.sub.6haloalkyl),
--OS(O).sub.2-(para-methylphenyl), or --OC(O)CF.sub.3; [0316]
R.sup.2, R.sup.3, and R.sup.5 each represent independently for each
occurrence hydrogen or methyl; [0317] R.sup.4 is hydrogen or
C.sub.1-C.sub.6alkyl; and [0318] t is 1, 2, or 3.
[0319] In certain embodiments, the therapeutic agent is an
organonitro compound embraced by Formula II-A as defined by
particular definitions for variables in Formula II-A, such as where
A is --N(R.sup.5)--. In certain other embodiments, A is
--N(CH.sub.3)--. In certain other embodiments, A is
--C(R.sup.2)(R.sup.3)--. In certain other embodiments, A is
--CH.sub.2--.
[0320] In certain embodiments, R.sup.1 is chloro. In certain other
embodiments, R.sup.1 is bromo. In certain embodiments, R.sup.1 is
--OS(O).sub.2--(C.sub.1-C.sub.6alkyl),
--OS(O).sub.2--(C.sub.1-C.sub.6haloalkyl), or
--OS(O).sub.2-(para-methylphenyl). In certain other embodiments,
R.sup.1 is --OS(O).sub.2CH.sub.3, --OS(O).sub.2CF.sub.3, or
--OS(O).sub.2-(para-methylphenyl). In certain other embodiments,
R.sup.1 is --OC(O)CF.sub.3.
[0321] In certain embodiments, R.sup.2 and R.sup.3 are
hydrogen.
[0322] In certain embodiments, R.sup.4 is hydrogen, methyl, ethyl,
propyl, butyl, or pentyl. In certain other embodiments, R.sup.4 is
methyl, ethyl or propyl. In certain other embodiments, R.sup.4 is
methyl.
[0323] In certain embodiments, R.sup.5 is hydrogen or methyl. In
certain other embodiments, R.sup.5 is hydrogen.
[0324] The description above describes multiple embodiments
relating to compounds of Formula II-A. The patent application
specifically contemplates all combinations of the embodiments. For
example, the invention contemplates a compound of Formula II-A
wherein A is --N(R.sup.5)--, and R.sup.2 and R.sup.3 are
hydrogen.
[0325] In certain embodiments, the therapeutic agent is a
hemoglobin conjugate of Formula III, and said therapeutic agent is
administered by injection (such as intravenous injection).
[0326] In certain embodiments, A.sup.1 is --C(O)-- and A.sup.2 is
N.
[0327] In certain embodiments, n is 0, and m is 2.
[0328] In certain embodiments, the therapeutic agent is
##STR00145##
or a pharmaceutically acceptable salt thereof, where z is an
integer from 1 to 10.
[0329] The therapeutic agents of Formulae I and II can be prepared
based on the procedures described in Schemes 1-9 above. The
hemoglobin conjugates of Formulae III and IV can be prepared by
admixing hemoglobin and a therapeutic agent of Formulae I and II,
respectively, to form the hemoglobin conjugate. In certain
embodiments, the beta-cysteine-93 residue of hemoglobin reacts with
the therapeutic agents of Formulae I and II form a thioether bond
due to reaction of the thiol group of the beta-cysteine-93 residue
of hemoglobin with the carbon atom bearing the R.sup.1 group in
Formulae I and II.
V. TREATING PATIENTS WITH ANEMIA
[0330] One aspect of the invention provides a method of treating a
patient suffering from anemia. The method comprises administering
to a patient in need thereof a therapeutic agent selected from the
group consisting of an organonitro compound of Formula I,
organonitro compound of Formula II, hemoglobin conjugate of Formula
III, hemoglobin conjugate of Formula IV, and an erythrocyte cell
that has been exposed to an organonitro compound of Formula I or
II; wherein Formula I is represented by:
##STR00146##
[0331] or a pharmaceutically acceptable salt or solvate thereof,
wherein: [0332] A.sup.1 is --C(O)-- or
--(C(R.sup.3).sub.2).sub.xC(O)(C(R.sup.3).sub.2).sub.x--; [0333]
A.sup.2 is N or --C(R.sup.4)--; [0334] R.sup.1 is halogen,
--OS(O).sub.2R.sup.5, or --OC(O)CF.sub.3; [0335] R.sup.2 is
C.sub.1-C.sub.6alkyl; [0336] R.sup.3 and R.sup.4 each represent
independently for each occurrence hydrogen or C.sub.1-C.sub.5alkyl;
[0337] R.sup.5 is C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6haloalkyl,
aryl, or aralkyl; [0338] m and p are independently 1, 2, or 3; and
[0339] n and x each represent independently for each occurrence 0,
1, 2, or 3;
[0340] Formula II is represented by:
##STR00147##
[0341] or a pharmaceutically acceptable salt or solvate thereof:
wherein: [0342] A.sup.1 is --C(O)-- or
--(C(R.sup.5).sub.2).sub.xC(O)(C(R.sup.5).sub.2).sub.x--; [0343]
A.sup.2 is --N(R.sup.5)-- or --C(R.sup.2)(R.sup.3)--; [0344]
R.sup.1 is halogen, --OS(O).sub.2R.sup.6, or --OC(O)CF.sub.3;
[0345] R.sup.2 and R.sup.3 each represent independently for each
occurrence hydrogen or C.sub.1-C.sub.6alkyl; or [0346] R.sup.2 and
R.sup.3 are taken together with the carbon atom to which they are
attached to form a 3-6 membered, saturated carbocyclic ring;
[0347] R.sup.4 is hydrogen or C.sub.1-C.sub.6alkyl; [0348] R.sup.5
represents independently for each occurrence hydrogen or
C.sub.1-C.sub.6alkyl; [0349] R.sup.6 is C.sub.1-C.sub.6alkyl,
C.sub.1-C.sub.6haloalkyl, aryl, or aralkyl; [0350] t is an integer
in the range from 1 to 12; and [0351] x represents independently
for each occurrence 0, 1, 2, or 3;
[0352] Formula III is represented by:
##STR00148##
[0353] or a pharmaceutically acceptable salt or solvate thereof,
wherein: [0354] A.sup.1 is --C(O)-- or
--C(O)(C(R.sup.3).sub.2).sub.x--; [0355] A.sup.2 is N or
--C(R.sup.4)--; [0356] R.sup.2 is C.sub.1-C.sub.6alkyl; [0357]
R.sup.3 and R.sup.4 each represent independently for each
occurrence hydrogen or C.sub.1-C.sub.5alkyl; [0358] m and p are
independently 1, 2, or 3; [0359] n is 0, 1, 2, or 3; [0360] x is 1,
2, or 3; and [0361] z is an integer from 1 to 10; and
[0362] Formula IV is represented by:
##STR00149##
[0363] or a pharmaceutically acceptable salt or solvate thereof,
wherein: [0364] A.sup.1 is --C(O)-- or
--C(O)(C(R.sup.5).sub.2).sub.x--; [0365] A.sup.2 is --N(R.sup.5)--
or --C(R.sup.2)(R.sup.3)--; [0366] R.sup.2 and R.sup.3 each
represent independently for each occurrence hydrogen or
C.sub.1-C.sub.6alkyl; or [0367] R.sup.2 and R.sup.3 are taken
together with the carbon atom to which they are attached to form a
3-6 membered, saturated carbocyclic ring; [0368] R.sup.4 is
hydrogen or C.sub.1-C.sub.6alkyl; [0369] R.sup.5 represents
independently for each occurrence hydrogen or C.sub.1-C.sub.6alkyl;
[0370] t is an integer in the range from 1 to 12; [0371] x is 1, 2,
or 3; and [0372] z is an integer from 1 to 10.
[0373] In certain embodiments, the method further comprises
administering a blood product to the patient by injection (such as
intravenous injection).
[0374] In certain embodiments, the blood product comprises
erythrocyte cells. In certain other embodiments, the blood product
comprises blood plasma. In certain embodiments, the blood product
comprises erythrocyte cells and blood plasma.
[0375] In certain embodiments, the blood product and organonitro
compound are administered to the patient concurrently.
[0376] In certain other embodiments, the blood product is
administered to the patient separately from the therapeutic
agent.
[0377] In certain embodiments, the patient receives, by intravenous
injection, a single composition comprising blood product and the
therapeutic agent. In certain other embodiments, the patient
receives, by intravenous injection, a single composition comprising
a therapeutic agent, plasma, and erythrocyte cells. One exemplary
composition is provided below in Table 2.
TABLE-US-00005 TABLE 2 Exemplary Composition for Intravenous
Injection* Component Amount Erythrocyte cells (vol %) 35-60 Plasma
(mL) 17 Anticoagulant As needed (e.g., 4 mL) Therapeutic Agent
(e.g., As needed, such as, an ABDNAZ) amount to treat hemorrhagic
shock. *Amounts are based on a composition having a total volume of
282 mL.
[0378] In certain embodiments, the method further comprises
administering an alkali metal nitrite to the patient. In other
embodiments, the method further comprises administering sodium
nitrite to the patient.
[0379] In certain embodiments, the therapeutic agent is an
organonitro compound of Formula I. In certain other embodiments,
the therapeutic agent is an erythrocyte cell that has been exposed
to an organonitro compound of Formula I, and said therapeutic agent
is administered by injection (such as intravenous injection).
[0380] In certain embodiments, A.sup.1 is --C(O)--, and A.sup.2 is
N.
[0381] In certain embodiments, R.sup.1 is bromo.
[0382] In certain embodiments, n is 0, and m is 2.
[0383] The description above describes multiple embodiments
relating to compounds of Formula I. The patent application
specifically contemplates all combinations of the embodiments. For
example, the invention contemplates a compound of Formula I wherein
A.sup.1 is --C(O)--, A.sup.2 is N, R.sup.1 is halogen, and n is
0.
[0384] In certain embodiments, the therapeutic agent is a compound
of Formula I-A:
##STR00150##
[0385] or a pharmaceutically acceptable salt or solvate thereof,
wherein: [0386] A is N or C(H); [0387] R.sup.1 is chloro, bromo,
--OS(O).sub.2--(C.sub.1-C.sub.6alkyl),
--OS(O).sub.2--(C.sub.1-C.sub.6haloalkyl),
--OS(O).sub.2-(para-methylphenyl), or --OC(O)CF.sub.3; [0388]
R.sup.2 represents independently for each occurrence hydrogen or
methyl; and y represents independently for each occurrence 1 or
2.
[0389] In certain embodiments, the therapeutic agent is compound
embraced by Formula I-A as defined by particular definitions for
variables in Formula I-A, such as where A is N. In certain other
embodiments, A is C(H).
[0390] In certain embodiments, R.sup.1 is chloro or bromo. In
certain embodiments, R.sup.1 is chloro. In certain other
embodiments, R.sup.1 is bromo. In certain embodiments, R.sup.1 is
--OS(O).sub.2--(C.sub.1-C.sub.6alkyl),
--OS(O).sub.2--(C.sub.1-C.sub.6haloalkyl), or
--OS(O).sub.2-(para-methylphenyl). In certain other embodiments,
R.sup.1 is --OS(O).sub.2CH.sub.3, --OS(O).sub.2CF.sub.3, or
--OS(O).sub.2-(para-methylphenyl). In certain other embodiments,
R.sup.1 is --OC(O)CF.sub.3.
[0391] In certain embodiments, R.sup.2 is hydrogen or methyl. In
certain embodiments, R.sup.2 is hydrogen.
[0392] In certain embodiments, y is 1. In certain embodiments, one
occurrence of y is 1, and the other occurrence of y is 2. In
certain other embodiments, y is 2.
[0393] The description above describes multiple embodiments
relating to compounds of Formula I-A. The patent application
specifically contemplates all combinations of the embodiments. For
example, the invention contemplates a compound of Formula I-A
wherein A is N, R.sup.1 is chloro or bromo, and R.sup.2 is
hydrogen.
[0394] In certain embodiments, the therapeutic agent is
##STR00151##
or a pharmaceutically acceptable salt thereof.
[0395] In certain embodiments, the therapeutic agent an organonitro
compound embraced by Formula II as defined by particular
definitions for variables in Formula II, such as where A.sup.1 is
--C(O)--. In certain other embodiments, A.sup.1 is
--(C(R.sup.5).sub.2).sub.xC(O)(C(R.sup.5).sub.2).sub.x--. In
certain other embodiments, A.sup.1 is
--C(O)(C(R.sup.5).sub.2).sub.x--.
[0396] In certain embodiments, A.sup.2 is --N(R.sup.5)--. In
certain other embodiments, A.sup.2 is --C(R.sup.2)(R.sup.3)--.
[0397] In certain embodiments, R.sup.1 is halogen. In certain other
embodiments, R.sup.1 is --OS(O).sub.2R.sup.6. In certain other
embodiments, R.sup.1 is --OC(O)CF.sub.3. In certain other
embodiments, R.sup.1 is chloro, bromo,
--OS(O).sub.2-(para-methylphenyl), --OS(O).sub.2CH.sub.3,
--OS(O).sub.2CF.sub.3, or --OC(O)CF.sub.3. In certain embodiments,
R.sup.1 is bromo.
[0398] In certain embodiments, R.sup.2 and R.sup.3 each represent
independently for each occurrence hydrogen or C.sub.1-C.sub.6alkyl.
In certain other embodiments, R.sup.2 and R.sup.3 each represent
independently for each occurrence hydrogen, methyl, ethyl, or
propyl. In certain other embodiments, R.sup.2 and R.sup.3 each
represent independently for each occurrence hydrogen or methyl. In
certain embodiments, R.sup.2 and R.sup.3 are hydrogen.
[0399] In certain embodiments, R.sup.4 is hydrogen, methyl, ethyl,
propyl, butyl, or pentyl. In certain other embodiments, R.sup.4 is
methyl, ethyl or propyl. In certain other embodiments, R.sup.4 is
methyl.
[0400] In certain embodiments, R.sup.5 is hydrogen or methyl. In
certain other embodiments, R.sup.5 is hydrogen.
[0401] In certain embodiments, R.sup.6 is C.sub.1-C.sub.6alkyl or
C.sub.1-C.sub.6haloalkyl. In certain other embodiments, R.sup.6 is
methyl, ethyl, or trifluoromethyl. In certain other embodiments,
R.sup.6 is aryl, such as phenyl.
[0402] In certain embodiments, t is 1, 2, 3, 4, 5 or 6. In certain
other embodiments, t is 1, 2, or 3. In certain other embodiments, t
is 1. In certain embodiments, x is 1 or 2.
[0403] The description above describes multiple embodiments
relating to compounds of Formula II. The patent application
specifically contemplates all combinations of the embodiments. For
example, the invention contemplates a compound of Formula II
wherein A.sup.1 is --C(O)--, A.sup.2 is --N(R.sup.5)--, and R.sup.2
and R.sup.3 are hydrogen.
[0404] In certain embodiments, the therapeutic agent is a compound
of Formula II-A:
##STR00152##
[0405] or a pharmaceutically acceptable salt or solvate thereof:
wherein:
[0406] A is --N(R.sup.5)-- or --C(R.sup.2)(R.sup.3)--;
[0407] R.sup.1 is chloro, bromo,
--OS(O).sub.2--(C.sub.1-C.sub.6alkyl),
--OS(O).sub.2--(C.sub.1-C.sub.6haloalkyl),
--OS(O).sub.2-(para-methylphenyl), or --OC(O)CF.sub.3;
[0408] R.sup.2, R.sup.3, and R.sup.5 each represent independently
for each occurrence hydrogen or methyl;
[0409] R.sup.4 is hydrogen or C.sub.1-C.sub.6alkyl; and
[0410] t is 1, 2, or 3.
[0411] In certain embodiments, the therapeutic agent is an
organonitro compound embraced by Formula II-A as defined by
particular definitions for variables in Formula II-A, such as where
A is --N(R.sup.5)--. In certain other embodiments, A is
--N(CH.sub.3)--. In certain other embodiments, A is
--C(R.sup.2)(R.sup.3)--. In certain other embodiments, A is
--CH.sub.2--.
[0412] In certain embodiments, R.sup.1 is chloro. In certain other
embodiments, R.sup.1 is bromo. In certain embodiments, R.sup.1 is
--OS(O).sub.2--(C.sub.1-C.sub.6alkyl),
--OS(O).sub.2--(C.sub.1-C.sub.6haloalkyl), or --OS(O).sub.2--
(para-methylphenyl). In certain other embodiments, R.sup.1 is
--OS(O).sub.2CH.sub.3, --OS(O).sub.2CF.sub.3, or
--OS(O).sub.2-(para-methylphenyl). In certain other embodiments,
R.sup.1 is --OC(O)CF.sub.3.
[0413] In certain embodiments, R.sup.2 and R.sup.3 are
hydrogen.
[0414] In certain embodiments, R.sup.4 is hydrogen, methyl, ethyl,
propyl, butyl, or pentyl. In certain other embodiments, R.sup.4 is
methyl, ethyl or propyl. In certain other embodiments, R.sup.4 is
methyl.
[0415] In certain embodiments, R.sup.5 is hydrogen or methyl. In
certain other embodiments, R.sup.5 is hydrogen.
[0416] The description above describes multiple embodiments
relating to compounds of Formula II-A. The patent application
specifically contemplates all combinations of the embodiments. For
example, the invention contemplates a compound of Formula II-A
wherein A is --N(R.sup.5)--, and R.sup.2 and R.sup.3 are
hydrogen.
[0417] In certain embodiments, the therapeutic agent is a
hemoglobin conjugate of Formula III, and said therapeutic agent is
administered by injection (such as intravenous injection).
[0418] In certain embodiments, A.sup.1 is --C(O)-- and A.sup.2 is
N.
[0419] In certain embodiments, n is 0, and m is 2.
[0420] In certain embodiments, the therapeutic agent is
##STR00153##
or a pharmaceutically acceptable salt thereof, where z is an
integer from 1 to 10.
[0421] The therapeutic agents of Formulae I and II can be prepared
based on the procedures described in Schemes 1-9 above. The
hemoglobin conjugates of Formulae III and IV can be prepared by
admixing hemoglobin and a therapeutic agent of Formulae I and II,
respectively, to form the hemoglobin conjugate. In certain
embodiments, the beta-cysteine-93 residue of hemoglobin reacts with
the therapeutic agents of Formulae I and II form a thioether bond
due to reaction of the thiol group of the beta-cysteine-93 residue
of hemoglobin with the carbon atom bearing the R.sup.1 group in
Formulae I and II.
VI. PRESERVING BLOOD PRODUCTS
[0422] One aspect of the invention provides a method of preserving
an isolated blood product. The method comprises exposing the
isolated blood product to an agent selected from the group
consisting of an organonitro compound of Formula I, organonitro
compound of Formula II, hemoglobin conjugate of Formula III,
hemoglobin conjugate of Formula IV, and an erythrocyte cell that
has been exposed to an organonitro compound of Formula I or II,
wherein Formula I is represented by:
##STR00154##
[0423] or a pharmaceutically acceptable salt or solvate thereof,
wherein:
[0424] A.sup.1 is --C(O)-- or
--(C(R.sup.3).sub.2).sub.xC(O)(C(R.sup.3).sub.2).sub.x--;
[0425] A.sup.2 is N or --C(R.sup.4)--;
[0426] R.sup.1 is halogen, --OS(O).sub.2R.sup.5, or
--OC(O)CF.sub.3;
[0427] R.sup.2 is C.sub.1-C.sub.6alkyl;
[0428] R.sup.3 and R.sup.4 each represent independently for each
occurrence hydrogen or C.sub.1-C.sub.5alkyl;
[0429] R.sup.5 is C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6haloalkyl,
aryl, or aralkyl;
[0430] m and p are independently 1, 2, or 3; and
[0431] n and x each represent independently for each occurrence 0,
1, 2, or 3;
[0432] Formula II is represented by:
##STR00155##
[0433] or a pharmaceutically acceptable salt or solvate thereof:
wherein:
[0434] A.sup.1 is --C(O)-- or
--(C(R.sup.5).sub.2).sub.xC(O)(C(R.sup.5).sub.2).sub.x--;
[0435] A.sup.2 is --N(R.sup.5)-- or --C(R.sup.2)(R.sup.3)--;
[0436] R.sup.1 is halogen, --OS(O).sub.2R.sup.6, or
--OC(O)CF.sub.3;
[0437] R.sup.2 and R.sup.3 each represent independently for each
occurrence hydrogen or C.sub.1-C.sub.6alkyl; or R.sup.2 and R.sup.3
are taken together with the carbon atom to which they are attached
to form a 3-6 membered, saturated carbocyclic ring;
[0438] R.sup.4 is hydrogen or C.sub.1-C.sub.6alkyl;
[0439] R.sup.5 represents independently for each occurrence
hydrogen or C.sub.1-C.sub.6alkyl;
[0440] R.sup.6 is C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6haloalkyl,
aryl, or aralkyl;
[0441] t is an integer in the range from 1 to 12; and
[0442] x represents independently for each occurrence 0, 1, 2, or
3;
[0443] Formula III is represented by:
##STR00156##
[0444] or a pharmaceutically acceptable salt or solvate thereof,
wherein:
[0445] A.sup.1 is --C(O)-- or --C(O)(C(R.sup.3).sub.2).sub.x--;
[0446] A.sup.2 is N or --C(R.sup.4)--;
[0447] R.sup.2 is C.sub.1-C.sub.6alkyl;
[0448] R.sup.3 and R.sup.4 each represent independently for each
occurrence hydrogen or C.sub.1-C.sub.5alkyl;
[0449] m and p are independently 1, 2, or 3;
[0450] n is 0, 1, 2, or 3;
[0451] x is 1, 2, or 3; and
[0452] z is an integer from 1 to 10; and
[0453] Formula IV is represented by:
##STR00157## [0454] or a pharmaceutically acceptable salt or
solvate thereof, wherein: [0455] A.sup.1 is --C(O)-- or
--C(O)(C(R.sup.5).sub.2).sub.x--; [0456] A.sup.2 is --N(R.sup.5)--
or --C(R.sup.2)(R.sup.3)--;
[0457] R.sup.2 and R.sup.3 each represent independently for each
occurrence hydrogen or C.sub.1-C.sub.6alkyl; or R.sup.2 and R.sup.3
are taken together with the carbon atom to which they are attached
to form a 3-6 membered, saturated carbocyclic ring;
[0458] R.sup.4 is hydrogen or C.sub.1-C.sub.6alkyl;
[0459] R.sup.5 represents independently for each occurrence
hydrogen or C.sub.1-C.sub.6alkyl;
[0460] t is an integer in the range from 1 to 12;
[0461] x is 1, 2, or 3; and
[0462] z is an integer from 1 to 10.
[0463] In certain embodiments, the isolated blood product is whole
blood. In certain embodiments, the isolated blood product comprises
erythrocyte cells. In certain other embodiments, the isolated blood
product is erythrocyte cells.
[0464] In certain embodiments, the method further comprises
exposing the isolated blood product to an alkali metal nitrite. In
other further embodiments, the method comprises exposing the
isolated blood product to sodium nitrite.
[0465] In certain embodiments, the agent is an organonitro compound
of Formula I. In certain embodiments, the agent is an erythrocyte
cell that has been exposed to an organonitro compound of Formula
I.
[0466] In certain embodiments, A.sup.1 is --C(O)--, and A.sup.2 is
N.
[0467] In certain embodiments, R.sup.1 is bromo.
[0468] In certain embodiments, n is 0, and m is 2.
[0469] The description above describes multiple embodiments
relating to compounds of Formula I. The patent application
specifically contemplates all combinations of the embodiments. For
example, the invention contemplates a compound of Formula I wherein
A.sup.1 is --C(O)--, A.sup.2 is N, R.sup.1 is halogen, and n is
0.
[0470] In certain embodiments, the agent is a compound of Formula
I-A:
##STR00158## [0471] or a pharmaceutically acceptable salt or
solvate thereof, wherein: [0472] A is N or C(H); [0473] R.sup.1 is
chloro, bromo, --OS(O).sub.2--(C.sub.1-C.sub.6alkyl),
--OS(O).sub.2--(C.sub.1-C.sub.6haloalkyl),
--OS(O).sub.2-(para-methylphenyl), or --OC(O)CF.sub.3; [0474]
R.sup.2 represents independently for each occurrence hydrogen or
methyl; and [0475] y represents independently for each occurrence 1
or 2.
[0476] In certain embodiments, the agent is compound embraced by
Formula I-A as defined by particular definitions for variables in
Formula I-A, such as where A is N. In certain other embodiments, A
is C(H).
[0477] In certain embodiments, R.sup.1 is chloro or bromo. In
certain embodiments, R.sup.1 is chloro. In certain other
embodiments, R.sup.1 is bromo. In certain embodiments, R.sup.1 is
--OS(O).sub.2--(C.sub.1-C.sub.6alkyl),
--OS(O).sub.2--(C.sub.1-C.sub.6haloalkyl), or
--OS(O).sub.2-(para-methylphenyl). In certain other embodiments,
R.sup.1 is --OS(O).sub.2CH.sub.3, --OS(O).sub.2CF.sub.3, or
--OS(O).sub.2-(para-methylphenyl). In certain other embodiments,
R.sup.1 is --OC(O)CF.sub.3.
[0478] In certain embodiments, R.sup.2 is hydrogen or methyl. In
certain embodiments, R.sup.2 is hydrogen.
[0479] In certain embodiments, y is 1. In certain embodiments, one
occurrence of y is 1, and the other occurrence of y is 2. In
certain other embodiments, y is 2.
[0480] The description above describes multiple embodiments
relating to compounds of Formula I-A. The patent application
specifically contemplates all combinations of the embodiments. For
example, the invention contemplates a compound of Formula I-A
wherein A is N, R.sup.1 is chloro or bromo, and R.sup.2 is
hydrogen.
[0481] In certain embodiments, the agent is
##STR00159##
or a pharmaceutically acceptable salt thereof.
[0482] In certain embodiments, the agent an organonitro compound
embraced by Formula II as defined by particular definitions for
variables in Formula II, such as where A.sup.1 is --C(O)--. In
certain other embodiments, A.sup.1 is
--(C(R.sup.5).sub.2).sub.xC(O)(C(R.sup.5).sub.2).sub.x--. In
certain other embodiments, A.sup.1 is
--C(O)(C(R.sup.5).sub.2).sub.x--.
[0483] In certain embodiments, A.sup.2 is --N(R.sup.5)--. In
certain other embodiments, A.sup.2 is --C(R.sup.2)(R.sup.3)--.
[0484] In certain embodiments, R.sup.1 is halogen. In certain other
embodiments, R.sup.1 is --OS(O).sub.2R.sup.6. In certain other
embodiments, R.sup.1 is --OC(O)CF.sub.3. In certain other
embodiments, R.sup.1 is chloro, bromo,
--OS(O).sub.2-(para-methylphenyl), --OS(O).sub.2CH.sub.3,
--OS(O).sub.2CF.sub.3, or --OC(O)CF.sub.3. In certain embodiments,
R.sup.1 is bromo.
[0485] In certain embodiments, R.sup.2 and R.sup.3 each represent
independently for each occurrence hydrogen or C.sub.1-C.sub.6alkyl.
In certain other embodiments, R.sup.2 and R.sup.3 each represent
independently for each occurrence hydrogen, methyl, ethyl, or
propyl. In certain other embodiments, R.sup.2 and R.sup.3 each
represent independently for each occurrence hydrogen or methyl. In
certain embodiments, R.sup.2 and R.sup.3 are hydrogen.
[0486] In certain embodiments, R.sup.4 is hydrogen, methyl, ethyl,
propyl, butyl, or pentyl. In certain other embodiments, R.sup.4 is
methyl, ethyl or propyl. In certain other embodiments, R.sup.4 is
methyl.
[0487] In certain embodiments, R.sup.5 is hydrogen or methyl. In
certain other embodiments, R.sup.5 is hydrogen.
[0488] In certain embodiments, R.sup.6 is C.sub.1-C.sub.6alkyl or
C.sub.1-C.sub.6haloalkyl. In certain other embodiments, R.sup.6 is
methyl, ethyl, or trifluoromethyl. In certain other embodiments,
R.sup.6 is aryl, such as phenyl.
[0489] In certain embodiments, t is 1, 2, 3, 4, 5 or 6. In certain
other embodiments, t is 1, 2, or 3. In certain other embodiments, t
is 1. In certain embodiments, x is 1 or 2.
[0490] The description above describes multiple embodiments
relating to compounds of Formula II. The patent application
specifically contemplates all combinations of the embodiments. For
example, the invention contemplates a compound of Formula II
wherein A.sup.1 is --C(O)--, A.sup.2 is --N(R.sup.5)--, and R.sup.2
and R.sup.3 are hydrogen.
[0491] In certain embodiments, the agent is a compound of Formula
II-A:
##STR00160##
[0492] or a pharmaceutically acceptable salt or solvate thereof:
wherein:
[0493] A is --N(R.sup.5)-- or --C(R.sup.2)(R.sup.3)--;
[0494] R.sup.1 is chloro, bromo,
--OS(O).sub.2--(C.sub.1-C.sub.6alkyl),
--OS(O).sub.2--(C.sub.1-C.sub.6haloalkyl),
--OS(O).sub.2-(para-methylphenyl), or --OC(O)CF.sub.3;
[0495] R.sup.2, R.sup.3, and R.sup.5 each represent independently
for each occurrence hydrogen or methyl;
[0496] R.sup.4 is hydrogen or C.sub.1-C.sub.6alkyl; and
[0497] t is 1, 2, or 3.
[0498] In certain embodiments, the agent is an organonitro compound
embraced by Formula II-A as defined by particular definitions for
variables in Formula II-A, such as where A is --N(R.sup.5)--. In
certain other embodiments, A is --N(CH.sub.3)--. In certain other
embodiments, A is --C(R.sup.2)(R.sup.3)--. In certain other
embodiments, A is --CH.sub.2--.
[0499] In certain embodiments, R.sup.1 is chloro. In certain other
embodiments, R.sup.1 is bromo. In certain embodiments, R.sup.1 is
--OS(O).sub.2--(C.sub.1-C.sub.6alkyl),
--OS(O).sub.2--(C.sub.1-C.sub.6haloalkyl), or
--OS(O).sub.2-(para-methylphenyl). In certain other embodiments,
R.sup.1 is --OS(O).sub.2CH.sub.3, --OS(O).sub.2CF.sub.3, or
--OS(O).sub.2-(para-methylphenyl). In certain other embodiments,
R.sup.1 is --OC(O)CF.sub.3.
[0500] In certain embodiments, R.sup.2 and R.sup.3 are
hydrogen.
[0501] In certain embodiments, R.sup.4 is hydrogen, methyl, ethyl,
propyl, butyl, or pentyl. In certain other embodiments, R.sup.4 is
methyl, ethyl or propyl. In certain other embodiments, R.sup.4 is
methyl.
[0502] In certain embodiments, R.sup.5 is hydrogen or methyl. In
certain other embodiments, R.sup.5 is hydrogen.
[0503] The description above describes multiple embodiments
relating to compounds of Formula II-A. The patent application
specifically contemplates all combinations of the embodiments. For
example, the invention contemplates a compound of Formula II-A
wherein A is --N(R.sup.5)--, and R.sup.2 and R.sup.3 are
hydrogen.
[0504] In certain embodiments, the agent is a hemoglobin conjugate
of Formula III.
[0505] In certain embodiments, A.sup.1 is --C(O)-- and A.sup.2 is
N.
[0506] In certain embodiments, n is 0, and m is 2.
[0507] In certain embodiments, the agent is
##STR00161##
or a pharmaceutically acceptable salt thereof, where z is an
integer from 1 to 10.
[0508] In certain embodiments, the agent is provided in an amount
effective to extend the storage life of the blood product by at
least 10%, 20% or 30% relative to the storage life of the blood
product without the agent. For example, in certain embodiments, the
agent is provided in an amount effective to extend the storage life
of the blood product by at least 1 day, 5 days, 10 days, or 15
days.
[0509] The agents of Formulae I and II can be prepared based on the
procedures described in Schemes 1-9 above. The hemoglobin
conjugates of Formulae III and IV can be prepared by admixing
hemoglobin and an agent of Formulae I and II, respectively, to form
the hemoglobin conjugate. In certain embodiments, the
beta-cysteine-93 residue of hemoglobin reacts with the agents of
Formulae I and II form a thioether bond due to reaction of the
thiol group of the beta-cysteine-93 residue of hemoglobin with the
carbon atom bearing the R.sup.1 group in Formulae I and II.
[0510] ABDNAZ and other compounds described herein are believed to
ameliorate the well-known storage lesion that occurs with stored
blood. Nitric oxide (NO) bioactivity of stored blood decreases
rapidly after blood is removed from the organism, which in part
limits the ability of stored blood to reverse arteriolar
vasoconstriction, capillary perfusion and tissue hypoxia. These
stresses consequently may affect the degree of intra and
extravascular hemolysis post-transfusion. Low levels of hemoglobin
(Hb) in plasma severely disrupt NO bioavailability by accelerating
NO dioxygenation reactions which results in decreased NO
concentration and leads to vasoconstriction. Restoration of NO
bioavailability prior or concurrently with the transfusion strategy
may therefore reduce the morbidity and mortality associated with
blood transfusion. Furthermore, enhancing the ability of blood to
generate NO by incubation with ABDNAZ or other compounds herein may
decrease the number of units of blood needed for treatment and
reduce healthcare costs while also extending the shelf life of
packed blood.
VII. ISOLATED BLOOD PRODUCT COMPOSITIONS
[0511] Another aspect of the invention provides an isolated blood
product composition. The composition comprises (i) a blood product,
and (ii) an agent selected from the group consisting of an
organonitro compound of Formula I, organonitro compound of Formula
II, hemoglobin conjugate of Formula III, hemoglobin conjugate of
Formula IV, and an erythrocyte cell that has been exposed to an
organonitro compound of Formula I or II; wherein Formula I is
represented by:
##STR00162##
[0512] or a pharmaceutically acceptable salt or solvate thereof,
wherein: [0513] A.sup.1 is --C(O)-- or
--(C(R.sup.3).sub.2).sub.xC(O)(C(R.sup.3).sub.2).sub.x--; [0514]
A.sup.2 is N or --C(R.sup.4)--; [0515] R.sup.1 is halogen,
--OS(O).sub.2R.sup.5, or --OC(O)CF.sub.3; [0516] R.sup.2 is
C.sub.1-C.sub.6alkyl; [0517] R.sup.3 and R.sup.4 each represent
independently for each occurrence hydrogen or C.sub.1-C.sub.5alkyl;
[0518] R.sup.5 is C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6haloalkyl,
aryl, or aralkyl; [0519] m and p are independently 1, 2, or 3; and
[0520] n and x each represent independently for each occurrence 0,
1, 2, or 3;
[0521] Formula II is represented by:
##STR00163##
[0522] or a pharmaceutically acceptable salt or solvate thereof:
wherein: [0523] A.sup.1 is --C(O)-- or
--(C(R.sup.5).sub.2).sub.xC(O)(C(R.sup.5).sub.2).sub.x--; [0524]
A.sup.2 is --N(R.sup.5)-- or --C(R.sup.2)(R.sup.3)--; [0525]
R.sup.1 is halogen, --OS(O).sub.2R.sup.6, or --OC(O)CF.sub.3;
[0526] R.sup.2 and R.sup.3 each represent independently for each
occurrence hydrogen or C.sub.1-C.sub.6alkyl; or R.sup.2 and R.sup.3
are taken together with the carbon atom to which they are attached
to form a 3-6 membered, saturated carbocyclic ring; [0527] R.sup.4
is hydrogen or C.sub.1-C.sub.6alkyl; [0528] R.sup.5 represents
independently for each occurrence hydrogen or C.sub.1-C.sub.6alkyl;
[0529] R.sup.6 is C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6haloalkyl,
aryl, or aralkyl; [0530] t is an integer in the range from 1 to 12;
and [0531] x represents independently for each occurrence 0, 1, 2,
or 3;
[0532] Formula III is represented by:
##STR00164##
[0533] or a pharmaceutically acceptable salt or solvate thereof,
wherein: [0534] A.sup.1 is --C(O)-- or
--C(O)(C(R.sup.3).sub.2).sub.x--; [0535] A.sup.2 is N or
--C(R.sup.4)--; [0536] R.sup.2 is C.sub.1-C.sub.6alkyl; [0537]
R.sup.3 and R.sup.4 each represent independently for each
occurrence hydrogen or C.sub.1-C.sub.5alkyl; [0538] m and p are
independently 1, 2, or 3; [0539] n is 0, 1, 2, or 3; [0540] x is 1,
2, or 3; and [0541] z is an integer from 1 to 10; and
[0542] Formula IV is represented by:
##STR00165##
[0543] or a pharmaceutically acceptable salt or solvate thereof,
wherein: [0544] A.sup.1 is --C(O)-- or
--C(O)(C(R.sup.5).sub.2).sub.x--; [0545] A.sup.2 is --N(R.sup.5)--
or --C(R.sup.2)(R.sup.3)--; [0546] R.sup.2 and R.sup.3 each
represent independently for each occurrence hydrogen or
C.sub.1-C.sub.6alkyl; or R.sup.2 and R.sup.3 are taken together
with the carbon atom to which they are attached to form a 3-6
membered, saturated carbocyclic ring; [0547] R.sup.4 is hydrogen or
C.sub.1-C.sub.6alkyl; [0548] R.sup.5 represents independently for
each occurrence hydrogen or C.sub.1-C.sub.6alkyl; [0549] t is an
integer in the range from 1 to 12; [0550] x is 1, 2, or 3; and
[0551] z is an integer from 1 to 10.
[0552] In certain embodiments, the blood product is whole blood. In
certain embodiments, the blood product comprises erythrocyte cells.
In certain other embodiments, the blood product comprises
erythrocyte cells and blood plasma. In certain embodiments, the
blood product is erythrocyte cells.
[0553] In certain embodiments, the composition further comprises an
alkali metal nitrite. In certain other embodiments, the composition
further comprises sodium nitrite.
[0554] The description above describes multiple embodiments
relating to compounds of Formula I. The patent application
specifically contemplates all combinations of the embodiments. For
example, the invention contemplates a compound of Formula I wherein
A.sup.1 is --C(O)--, A.sup.2 is N, R.sup.1 is halogen, and n is
0.
[0555] In certain embodiments, the agent is a compound of Formula
I-A:
##STR00166##
[0556] or a pharmaceutically acceptable salt or solvate thereof,
wherein: [0557] A is N or C(H); [0558] R.sup.1 is chloro, bromo,
--OS(O).sub.2--(C.sub.1-C.sub.6alkyl),
--OS(O).sub.2--(C.sub.1-C.sub.6haloalkyl),
--OS(O).sub.2-(para-methylphenyl), or --OC(O)CF.sub.3; [0559]
R.sup.2 represents independently for each occurrence hydrogen or
methyl; and y represents independently for each occurrence 1 or
2.
[0560] In certain embodiments, the agent is compound embraced by
Formula I-A as defined by particular definitions for variables in
Formula I-A, such as where A is N. In certain other embodiments, A
is C(H).
[0561] In certain embodiments, R.sup.1 is chloro or bromo. In
certain embodiments, R.sup.1 is chloro. In certain other
embodiments, R.sup.1 is bromo. In certain embodiments, R.sup.1 is
--OS(O).sub.2--(C.sub.1-C.sub.6alkyl),
--OS(O).sub.2--(C.sub.1-C.sub.6haloalkyl), or
--OS(O).sub.2-(para-methylphenyl). In certain other embodiments,
R.sup.1 is --OS(O).sub.2CH.sub.3, --OS(O).sub.2CF.sub.3, or
--OS(O).sub.2-(para-methylphenyl). In certain other embodiments,
R.sup.1 is --OC(O)CF.sub.3.
[0562] In certain embodiments, R.sup.2 is hydrogen or methyl. In
certain embodiments, R.sup.2 is hydrogen.
[0563] In certain embodiments, y is 1. In certain embodiments, one
occurrence of y is 1, and the other occurrence of y is 2. In
certain other embodiments, y is 2.
[0564] The description above describes multiple embodiments
relating to compounds of Formula I-A. The patent application
specifically contemplates all combinations of the embodiments. For
example, the invention contemplates a compound of Formula I-A
wherein A is N, R.sup.1 is chloro or bromo, and R.sup.2 is
hydrogen.
[0565] In certain embodiments, the agent is
##STR00167##
or a pharmaceutically acceptable salt thereof.
[0566] In certain embodiments, the agent an organonitro compound
embraced by Formula II as defined by particular definitions for
variables in Formula II, such as where A.sup.1 is --C(O)--. In
certain other embodiments, A.sup.1 is
--(C(R.sup.5).sub.2).sub.xC(O)(C(R.sup.5).sub.2).sub.x--. In
certain other embodiments, A.sup.1 is
--C(O)(C(R.sup.5).sub.2).sub.x--.
[0567] In certain embodiments, A.sup.2 is --N(R.sup.5)--. In
certain other embodiments, A.sup.2 is --C(R.sup.2)(R.sup.3)--.
[0568] In certain embodiments, R.sup.1 is halogen. In certain other
embodiments, R.sup.1 is --OS(O).sub.2R.sup.6. In certain other
embodiments, R.sup.1 is --OC(O)CF.sub.3. In certain other
embodiments, R.sup.1 is chloro, bromo,
--OS(O).sub.2-(para-methylphenyl), --OS(O).sub.2CH.sub.3,
--OS(O).sub.2CF.sub.3, or --OC(O)CF.sub.3. In certain embodiments,
R.sup.1 is bromo.
[0569] In certain embodiments, R.sup.2 and R.sup.3 each represent
independently for each occurrence hydrogen or C.sub.1-C.sub.6alkyl.
In certain other embodiments, R.sup.2 and R.sup.3 each represent
independently for each occurrence hydrogen, methyl, ethyl, or
propyl. In certain other embodiments, R.sup.2 and R.sup.3 each
represent independently for each occurrence hydrogen or methyl. In
certain embodiments, R.sup.2 and R.sup.3 are hydrogen.
[0570] In certain embodiments, R.sup.4 is hydrogen, methyl, ethyl,
propyl, butyl, or pentyl. In certain other embodiments, R.sup.4 is
methyl, ethyl or propyl. In certain other embodiments, R.sup.4 is
methyl.
[0571] In certain embodiments, R.sup.5 is hydrogen or methyl. In
certain other embodiments, R.sup.5 is hydrogen.
[0572] In certain embodiments, R.sup.6 is C.sub.1-C.sub.6alkyl or
C.sub.1-C.sub.6haloalkyl. In certain other embodiments, R.sup.6 is
methyl, ethyl, or trifluoromethyl. In certain other embodiments,
R.sup.6 is aryl, such as phenyl.
[0573] In certain embodiments, t is 1, 2, 3, 4, 5 or 6. In certain
other embodiments, t is 1, 2, or 3. In certain other embodiments, t
is 1. In certain embodiments, x is 1 or 2.
[0574] The description above describes multiple embodiments
relating to compounds of Formula II. The patent application
specifically contemplates all combinations of the embodiments. For
example, the invention contemplates a compound of Formula II
wherein A.sup.1 is --C(O)--, A.sup.2 is --N(R.sup.5)--, and R.sup.2
and R.sup.3 are hydrogen.
[0575] In certain embodiments, the agent is a compound of Formula
II-A:
##STR00168##
[0576] or a pharmaceutically acceptable salt or solvate thereof:
wherein:
[0577] A is --N(R.sup.5)-- or --C(R.sup.2)(R.sup.3)--;
[0578] R.sup.1 is chloro, bromo,
--OS(O).sub.2--(C.sub.1-C.sub.6alkyl),
--OS(O).sub.2--(C.sub.1-C.sub.6haloalkyl),
--OS(O).sub.2-(para-methylphenyl), or --OC(O)CF.sub.3;
[0579] R.sup.2, R.sup.3, and R.sup.5 each represent independently
for each occurrence hydrogen or methyl;
[0580] R.sup.4 is hydrogen or C.sub.1-C.sub.6alkyl; and
[0581] t is 1, 2, or 3.
[0582] In certain embodiments, the agent is an organonitro compound
embraced by Formula II-A as defined by particular definitions for
variables in Formula II-A, such as where A is --N(R.sup.5)--. In
certain other embodiments, A is --N(CH.sub.3)--. In certain other
embodiments, A is --C(R.sup.2)(R.sup.3)--. In certain other
embodiments, A is --CH.sub.2--.
[0583] In certain embodiments, R.sup.1 is chloro. In certain other
embodiments, R.sup.1 is bromo. In certain embodiments, R.sup.1 is
--OS(O).sub.2--(C.sub.1-C.sub.6alkyl),
--OS(O).sub.2--(C.sub.1-C.sub.6haloalkyl), or
--OS(O).sub.2-(para-methylphenyl). In certain other embodiments,
R.sup.1 is --OS(O).sub.2CH.sub.3, --OS(O).sub.2CF.sub.3, or
--OS(O).sub.2-(para-methylphenyl). In certain other embodiments,
R.sup.1 is --OC(O)CF.sub.3.
[0584] In certain embodiments, R.sup.2 and R.sup.3 are
hydrogen.
[0585] In certain embodiments, R.sup.4 is hydrogen, methyl, ethyl,
propyl, butyl, or pentyl. In certain other embodiments, R.sup.4 is
methyl, ethyl or propyl. In certain other embodiments, R.sup.4 is
methyl.
[0586] In certain embodiments, R.sup.5 is hydrogen or methyl. In
certain other embodiments, R.sup.5 is hydrogen.
[0587] The description above describes multiple embodiments
relating to compounds of Formula II-A. The patent application
specifically contemplates all combinations of the embodiments. For
example, the invention contemplates a compound of Formula II-A
wherein A is --N(R.sup.5)--, and R.sup.2 and R.sup.3 are
hydrogen.
[0588] In certain embodiments, the agent is a hemoglobin conjugate
of Formula III.
[0589] In certain embodiments, A.sup.1 is --C(O)-- and A.sup.2 is
N.
[0590] In certain embodiments, n is 0, and m is 2.
[0591] In certain embodiments, the agent is
##STR00169##
or a pharmaceutically acceptable salt thereof, where z is an
integer from 1 to 10.
[0592] The agents of Formulae I and II can be prepared based on the
procedures described in Schemes 1-9 above. The hemoglobin
conjugates of Formulae III and IV can be prepared by admixing
hemoglobin and an agent of Formulae I and II, respectively, to form
the hemoglobin conjugate. In certain embodiments, the
beta-cysteine-93 residue of hemoglobin reacts with the agents of
Formulae I and II form a thioether bond due to reaction of the
thiol group of the beta-cysteine-93 residue of hemoglobin with the
carbon atom bearing the R.sup.1 group in Formulae I and II.
[0593] In certain embodiments, the isolated blood product
composition comprises a compound of Formula II, plasma, and
erythrocyte cells. In certain other embodiments, the isolated blood
product composition has the features provided below in Table 3.
TABLE-US-00006 TABLE 3 Exemplary Isolated Blood Product
Composition* Component Amount Erythrocyte cells (vol %) 35-60
Plasma (mL) 17 Anticoagulant As needed (e.g., 4 mL) Therapeutic
Agent (e.g., As needed, such as, an ABDNAZ) amount to treat
hemorrhagic shock. *Amounts are based on a composition having a
total volume of 282 mL.
VIII. HEMOGLOBIN CONJUGATES
[0594] Another aspect of the invention provides an isolated
hemoglobin conjugate represented by Formula III or IV:
##STR00170##
[0595] or a pharmaceutically acceptable salt or solvate thereof,
wherein: [0596] A.sup.1 is --C(O)-- or
--C(O)(C(R.sup.3).sub.2).sub.x--; [0597] A.sup.2 is N or
--C(R.sup.4)--; [0598] R.sup.2 is C.sub.1-C.sub.6alkyl; [0599]
R.sup.3 and R.sup.4 each represent independently for each
occurrence hydrogen or C.sub.1-C.sub.5alkyl; [0600] m and p are
independently 1, 2, or 3; [0601] n is 0, 1, 2, or 3; [0602] x is 1,
2, or 3; and [0603] z is an integer from 1 to 10; and
[0604] Formula IV is represented by:
##STR00171##
[0605] or a pharmaceutically acceptable salt or solvate thereof,
wherein: [0606] A.sup.1 is --C(O)-- or
--C(O)(C(R.sup.5).sub.2).sub.x--; [0607] A.sup.2 is --N(R.sup.5)--
or --C(R.sup.2)(R.sup.3)--; [0608] R.sup.2 and R.sup.3 each
represent independently for each occurrence hydrogen or
C.sub.1-C.sub.6alkyl; or R.sup.2 and R.sup.3 are taken together
with the carbon atom to which they are attached to form a 3-6
membered, saturated carbocyclic ring; [0609] R.sup.4 is hydrogen or
C.sub.1-C.sub.6alkyl; [0610] R.sup.5 represents independently for
each occurrence hydrogen or C.sub.1-C.sub.6alkyl; [0611] t is an
integer in the range from 1 to 12; [0612] x is 1, 2, or 3; and
[0613] z is an integer from 1 to 10.
[0614] In certain embodiments, the isolated hemoglobin conjugate is
represented by Formula III.
[0615] In certain embodiments, A.sup.1 is --C(O)--, and A.sup.2 is
N.
[0616] In certain embodiments, n is 0, and m is 2.
[0617] In certain embodiments, the isolated hemoglobin conjugate
is
##STR00172##
or a pharmaceutically acceptable salt thereof, wherein z is an
integer from 1 to 10.
[0618] The isolated hemoglobin conjugates of Formulae III and IV
can be prepared by admixing hemoglobin and an agent of Formulae I
and II, respectively, to form the isolated hemoglobin conjugate. In
certain embodiments, the beta-cysteine-93 residue of hemoglobin
reacts with the agents of Formulae I and II form a thioether bond
due to reaction of the thiol group of the beta-cysteine-93 residue
of hemoglobin with the carbon atom bearing the R.sup.1 group in
Formulae I and II.
[0619] In certain embodiments, another aspect of the invention
provides a pharmaceutical composition. The composition comprises a
pharmaceutically acceptable carrier and an isolated hemoglobin
conjugate as described herein. In certain embodiments, the
pharmaceutically acceptable carrier comprises blood plasma.
IX. PHARMACEUTICAL COMPOSITIONS
[0620] The invention provides pharmaceutical compositions. As a
general matter, the pharmaceutical composition contains at least
one active agent and a pharmaceutically acceptable carrier. In
certain embodiments, the pharmaceutical compositions comprise an
inorganic nitrite salt and/or an allosteric modulator of hemoglobin
that promotes nitrite reductase activity. In certain other
embodiments, the pharmaceutical compositions preferably comprise a
therapeutically-effective amount of an inorganic nitrite salt
and/or an allosteric modulator of hemoglobin that promotes nitrite
reductase activity, formulated together with one or more
pharmaceutically acceptable carriers (additives) and/or diluents.
As described in detail below, the pharmaceutical compositions of
the present invention may be specially formulated for
administration in solid or liquid form, including those adapted for
the following: (1) oral administration, for example, drenches
(aqueous or non-aqueous solutions or suspensions), tablets (e.g.,
those targeted for buccal, sublingual, and/or systemic absorption),
boluses, powders, granules, pastes for application to the tongue;
(2) parenteral administration by, for example, subcutaneous,
intramuscular, intravenous or epidural injection as, for example, a
sterile solution or suspension, or sustained-release formulation;
(3) topical application, for example, as a cream, ointment, or a
controlled-release patch or spray applied to the skin; (4)
intravaginally or intrarectally, for example, as a pessary, cream
or foam; (5) sublingually; (6) ocularly; (7) transdermally; or (8)
nasally.
[0621] Wetting agents, emulsifiers and lubricants, such as sodium
lauryl sulfate and magnesium stearate, as well as coloring agents,
release agents, coating agents, sweetening, flavoring and perfuming
agents, preservatives and antioxidants can also be present in the
compositions.
[0622] Examples of pharmaceutically-acceptable antioxidants
include: (1) water soluble antioxidants, such as ascorbic acid,
cysteine hydrochloride, sodium bisulfate, sodium metabisulfite,
sodium sulfite and the like; (2) oil-soluble antioxidants, such as
ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated
hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol,
and the like; and (3) metal chelating agents, such as citric acid,
ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid,
phosphoric acid, and the like.
[0623] Formulations of the present invention include those suitable
for oral, nasal, topical (including buccal and sublingual), rectal,
vaginal and/or parenteral administration. The formulations may
conveniently be presented in unit dosage form and may be prepared
by any methods well known in the art of pharmacy. The amount of
active ingredient which can be combined with a carrier material to
produce a single dosage form will vary depending upon the host
being treated, the particular mode of administration. The amount of
active ingredient that can be combined with a carrier material to
produce a single dosage form will generally be that amount of the
compound which produces a therapeutic effect. Generally, out of one
hundred percent, this amount will range from about 0.1 percent to
about ninety-nine percent of active ingredient, preferably from
about 5 percent to about 70 percent, most preferably from about 10
percent to about 30 percent.
[0624] In certain embodiments, a formulation of the present
invention comprises an excipient selected from the group consisting
of cyclodextrins, celluloses, liposomes, micelle forming agents,
e.g., bile acids, and polymeric carriers, e.g., polyesters and
polyanhydrides; and a compound of the present invention. In certain
embodiments, an aforementioned formulation renders a compound of
the present invention orally bioavailable.
[0625] Methods of preparing these formulations or compositions
include the step of bringing into association a compound of the
present invention with the carrier and, optionally, one or more
accessory ingredients. In general, the formulations are prepared by
uniformly and intimately bringing into association a compound of
the present invention with liquid carriers, or finely divided solid
carriers, or both, and then, if necessary, shaping the product.
[0626] Formulations of the invention suitable for oral
administration may be in the form of capsules, cachets, pills,
tablets, lozenges (using a flavored basis, usually sucrose and
acacia or tragacanth), powders, granules, or as a solution or a
suspension in an aqueous or non-aqueous liquid, or as an
oil-in-water or water-in-oil liquid emulsion, or as an elixir or
syrup, or as pastilles (using an inert base, such as gelatin and
glycerin, or sucrose and acacia) and/or as mouth washes and the
like, each containing a predetermined amount of a compound of the
present invention as an active ingredient. A compound of the
present invention may also be administered as a bolus, electuary or
paste.
[0627] In solid dosage forms of the invention for oral
administration (capsules, tablets, pills, dragees, powders,
granules, trouches and the like), the active ingredient is mixed
with one or more pharmaceutically-acceptable carriers, such as
sodium citrate or dicalcium phosphate, and/or any of the following:
(1) fillers or extenders, such as starches, lactose, sucrose,
glucose, mannitol, and/or silicic acid; (2) binders, such as, for
example, carboxymethylcellulose, alginates, gelatin, polyvinyl
pyrrolidone, sucrose and/or acacia; (3) humectants, such as
glycerol; (4) disintegrating agents, such as agar-agar, calcium
carbonate, potato or tapioca starch, alginic acid, certain
silicates, and sodium carbonate; (5) solution retarding agents,
such as paraffin; (6) absorption accelerators, such as quaternary
ammonium compounds and surfactants, such as poloxamer and sodium
lauryl sulfate; (7) wetting agents, such as, for example, cetyl
alcohol, glycerol monostearate, and non-ionic surfactants; (8)
absorbents, such as kaolin and bentonite clay; (9) lubricants, such
as talc, calcium stearate, magnesium stearate, solid polyethylene
glycols, sodium lauryl sulfate, zinc stearate, sodium stearate,
stearic acid, and mixtures thereof; (10) coloring agents; and (11)
controlled release agents such as crospovidone or ethyl cellulose.
In the case of capsules, tablets and pills, the pharmaceutical
compositions may also comprise buffering agents. Solid compositions
of a similar type may also be employed as fillers in soft and
hard-shelled gelatin capsules using such excipients as lactose or
milk sugars, as well as high molecular weight polyethylene glycols
and the like.
[0628] A tablet may be made by compression or molding, optionally
with one or more accessory ingredients. Compressed tablets may be
prepared using binder (for example, gelatin or hydroxypropylmethyl
cellulose), lubricant, inert diluent, preservative, disintegrant
(for example, sodium starch glycolate or cross-linked sodium
carboxymethyl cellulose), surface-active or dispersing agent.
Molded tablets may be made by molding in a suitable machine a
mixture of the powdered compound moistened with an inert liquid
diluent.
[0629] The tablets, and other solid dosage forms of the
pharmaceutical compositions of the present invention, such as
dragees, capsules, pills and granules, may optionally be scored or
prepared with coatings and shells, such as enteric coatings and
other coatings well known in the pharmaceutical-formulating art.
They may also be formulated so as to provide slow or controlled
release of the active ingredient therein using, for example,
hydroxypropylmethyl cellulose in varying proportions to provide the
desired release profile, other polymer matrices, liposomes and/or
microspheres. They may be formulated for rapid release, e.g.,
freeze-dried. They may be sterilized by, for example, filtration
through a bacteria-retaining filter, or by incorporating
sterilizing agents in the form of sterile solid compositions which
can be dissolved in sterile water, or some other sterile injectable
medium immediately before use. These compositions may also
optionally contain opacifying agents and may be of a composition
that they release the active ingredient(s) only, or preferentially,
in a certain portion of the gastrointestinal tract, optionally, in
a delayed manner. Examples of embedding compositions which can be
used include polymeric substances and waxes. The active ingredient
can also be in micro-encapsulated form, if appropriate, with one or
more of the above-described excipients.
[0630] Liquid dosage forms for oral administration of the compounds
of the invention include pharmaceutically acceptable emulsions,
microemulsions, solutions, suspensions, syrups and elixirs. In
addition to the active ingredient, 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, oils (in particular,
cottonseed, groundnut, corn, germ, olive, castor and sesame oils),
glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty
acid esters of sorbitan, and mixtures thereof
[0631] Besides inert diluents, the oral compositions can also
include adjuvants such as wetting agents, emulsifying and
suspending agents, sweetening, flavoring, coloring, perfuming and
preservative agents.
[0632] Suspensions, in addition to the active compounds, may
contain suspending agents as, for example, ethoxylated isostearyl
alcohols, polyoxyethylene sorbitol and sorbitan esters,
microcrystalline cellulose, aluminum metahydroxide, bentonite,
agar-agar and tragacanth, and mixtures thereof
[0633] Formulations of the pharmaceutical compositions of the
invention for rectal or vaginal administration may be presented as
a suppository, which may be prepared by mixing one or more
compounds of the invention with one or more suitable nonirritating
excipients or carriers comprising, for example, cocoa butter,
polyethylene glycol, a suppository wax or a salicylate, and which
is solid at room temperature, but liquid at body temperature and,
therefore, will melt in the rectum or vaginal cavity and release
the active compound.
[0634] Dosage forms for the topical or transdermal administration
of a compound of this invention include powders, sprays, ointments,
pastes, creams, lotions, gels, solutions, patches and inhalants.
The active compound may be mixed under sterile conditions with a
pharmaceutically-acceptable carrier, and with any preservatives,
buffers, or propellants which may be required.
[0635] The ointments, pastes, creams and gels may contain, in
addition to an active compound of this invention, excipients, such
as animal and vegetable fats, oils, waxes, paraffins, starch,
tragacanth, cellulose derivatives, polyethylene glycols, silicones,
bentonites, silicic acid, talc and zinc oxide, or mixtures
thereof
[0636] Powders and sprays can contain, in addition to a compound of
this invention, excipients such as lactose, talc, silicic acid,
aluminum hydroxide, calcium silicates and polyamide powder, or
mixtures of these substances. Sprays can additionally contain
customary propellants, such as chlorofluorohydrocarbons and
volatile unsubstituted hydrocarbons, such as butane and
propane.
[0637] Transdermal patches have the added advantage of providing
controlled delivery of a compound of the present invention to the
body. Such dosage forms can be made by dissolving or dispersing the
compound in the proper medium. Absorption enhancers can also be
used to increase the flux of the compound across the skin. The rate
of such flux can be controlled by either providing a rate
controlling membrane or dispersing the compound in a polymer matrix
or gel.
[0638] Ophthalmic formulations, eye ointments, powders, solutions
and the like, are also contemplated as being within the scope of
this invention.
[0639] Pharmaceutical compositions of this invention suitable for
parenteral administration comprise one or more compounds of the
invention in combination with one or more
pharmaceutically-acceptable sterile isotonic aqueous or nonaqueous
solutions, dispersions, suspensions or emulsions, or sterile
powders which may be reconstituted into sterile injectable
solutions or dispersions just prior to use, which may contain
sugars, alcohols, antioxidants, buffers, bacteriostats, solutes
which render the formulation isotonic with the blood of the
intended recipient or suspending or thickening agents.
[0640] Examples of suitable aqueous and nonaqueous carriers which
may be employed in the pharmaceutical compositions of the invention
include water, ethanol, polyols (such as glycerol, propylene
glycol, polyethylene glycol, and the like), and suitable mixtures
thereof, vegetable oils, such as olive oil, and injectable organic
esters, such as ethyl oleate. Proper fluidity can be maintained,
for example, by the use of coating materials, such as lecithin, by
the maintenance of the required particle size in the case of
dispersions, and by the use of surfactants.
[0641] These compositions may also contain adjuvants such as
preservatives, wetting agents, emulsifying agents and dispersing
agents. Prevention of the action of microorganisms upon the subject
compounds may be ensured by the inclusion of various antibacterial
and antifungal agents, for example, paraben, chlorobutanol, phenol
sorbic acid, and the like. It may also be desirable to include
isotonic agents, such as sugars, sodium chloride, and the like into
the compositions. In addition, prolonged absorption of the
injectable pharmaceutical form may be brought about by the
inclusion of agents which delay absorption such as aluminum
monostearate and gelatin.
[0642] In some cases, in order to prolong the effect of a drug, it
is desirable to slow the absorption of the drug administered by
subcutaneous or intramuscular injection. This may be accomplished
by the use of a liquid suspension of crystalline or amorphous
material having poor water solubility. The rate of absorption of
the drug then depends upon its rate of dissolution which, in turn,
may depend upon crystal size and crystalline form. Alternatively,
delayed absorption of a parenterally-administered drug form is
accomplished by dissolving or suspending the drug in an oil
vehicle.
[0643] Injectable depot forms are made by forming microencapsule
matrices of the subject compounds in biodegradable polymers such as
polylactide-polyglycolide. Depending on the ratio of drug to
polymer, and the nature of the particular polymer employed, the
rate of drug release can be controlled. Examples of other
biodegradable polymers include poly(orthoesters) and
poly(anhydrides). Depot injectable formulations are also prepared
by entrapping the drug in liposomes or microemulsions which are
compatible with body tissue.
[0644] When the compounds of the present invention are administered
as pharmaceuticals, to humans and animals, they can be given per se
or as a pharmaceutical composition containing, for example, 0.1 to
99% (more preferably, 10 to 30%) of active ingredient in
combination with a pharmaceutically acceptable carrier.
[0645] The preparations of the present invention may be given
orally, parenterally, topically, or rectally. They are of course
given in forms suitable for each administration route. For example,
they are administered in tablets or capsule form, by injection,
inhalation, eye lotion, ointment, suppository, etc. administration
by injection, infusion or inhalation; topical by lotion or
ointment; and rectal by suppositories. Oral administrations are
preferred.
[0646] The phrase "parenteral administration" and "administered
parenterally" as used herein means modes of administration other
than enteral and topical administration, usually by injection, and
include, without limitation, intravenous, intramuscular,
intraarterial, intrathecal, intracapsular, intraorbital,
intracardiac, intradermal, intraperitoneal, transtracheal,
subcutaneous, subcuticular, intraarticulare, subcapsular,
subarachnoid, intraspinal, and intrasternal injection and
infusion.
[0647] The phrases "systemic administration," "administered
systemically," "peripheral administration" and "administered
peripherally" as used herein mean the administration of a compound,
drug or other material other than directly into the central nervous
system, such that it enters the patient's system and, thus, is
subject to metabolism and other like processes, for example,
subcutaneous administration.
[0648] These compounds may be administered to humans and other
animals for therapy by any suitable route of administration,
including orally, nasally, as by, for example, a spray, rectally,
intravaginally, parenterally, intracisternally and topically, as by
powders, ointments or drops, including buccally and
sublingually.
[0649] Regardless of the route of administration selected, the
compounds of the present invention, which may be used in a suitable
hydrated form, and/or the pharmaceutical compositions of the
present invention, are formulated into pharmaceutically-acceptable
dosage forms by conventional methods known to those of skill in the
art.
X. KITS FOR USE IN MEDICAL APPLICATIONS
[0650] Another aspect of the invention provides a kit for treating
a disorder. In certain embodiments, the kit comprises: (i) an
inorganic nitrite salt, (ii) a nitrite reductase promoter (which
preferably is an allosteric modulator of hemoglobin), and (iii)
instructions for using the kit to treat a medical disorder.
[0651] In certain embodiments, the disorder is cancer, a
cardiovascular disorder, an ischemic condition, a hemolytic
condition, or a bacterial infection. In certain other embodiments,
the disorder is cancer, such as a tumor. In certain other
embodiments, the disorder is a cardiovascular disorder, such as
pulmonary hypertension, systemic hypertension, angina, Cardiac
Syndrome X, myocardial infarction, peripheral artery disease, or
Raynaud's disease.
[0652] In certain embodiments, the allosteric modulator of
hemoglobin is one of the generic or specific an allosteric
modulators of hemoglobin described in Section II, such as a
compound of Formula I, a compound embraced by one of the further
embodiments describing definitions for certain variables of Formula
I, a compound of Formula I-A, or a compound embraced by one of the
further embodiments describing definitions for certain variables of
Formula I-A. In certain embodiments, the an allosteric modulator of
hemoglobin is a compound of Formula II, a compound embraced by one
of the further embodiments describing definitions for certain
variables of Formula II, a compound of Formula II-A, or a compound
embraced by one of the further embodiments describing definitions
for certain variables of Formula II-A.
[0653] The description above describes multiple aspects and
embodiments of the invention, including allosteric modulators of
hemoglobin, compositions comprising an allosteric modulator of
hemoglobin, methods of using the allosteric modulators of
hemoglobin in combination with an inorganic nitrite salt, and kits.
The patent application specifically contemplates all combinations
and permutations of the aspects and embodiments. For example, the
invention contemplates treating tumors in a human patient by
administering a therapeutically effective amount of sodium nitrite
in combination with an allosteric modulator of hemoglobin of
Formula I-A. Further, for example, the invention contemplates a kit
for treating tumors, the kit comprising (i) an inorganic nitrite
salt described herein, such as sodium nitrite, (ii) an allosteric
modulator of hemoglobin, such as a compound of Formula I, and (iii)
instructions for treating a tumor.
EXAMPLES
[0654] The invention now being generally described, will be more
readily understood by reference to the following examples, which
are included merely for purposes of illustration of certain aspects
and embodiments of the present invention, and are not intended to
limit the invention.
Example 1
[0655] The ability of ABDNAZ to increase nitric oxide production in
a sample of blood under aerobic conditions and anaerobic conditions
was evaluated. Experimental procedures and results are provided
below.
Experimental Procedures:
[0656] A 0.5 mL aliquot of blood was placed in a tonometer. ABDNAZ
(5 .mu.L of a 300 mM solution of ABDNAZ in dimethylsulfoxide) was
optionally added to the blood sample in the Tonometer. Sodium
nitrite was added to the blood sample to achieve a sodium nitrite
concentration of 167 mM. Gas (either air or N.sub.2) was passed
through the Tonometer at a flow rate of 150 mL/min. Gas exiting the
Tonometer was collected in three-minute intervals for a period of
thirty minutes. The amount of nitric oxide in each of the collected
fractions was measured by a chemiluminescent reaction with ozone in
a Sievers Nitric Oxide Analyzer. The amount of nitric oxide
produced from the blood sample was expressed per mole of hemoglobin
present in the blood sample. All experiments were performed at
37.degree. C.
[0657] The amount of hemoglobin (Hb) and hematocrit (Hct) in the
blood sample was determined using an Advia analyzer.
Results:
[0658] Experimental data showing the amount of nitric oxide
produced by the blood samples are shown in FIGS. 1-8. In
particular, FIGS. 1-5 show the cumulative amount of nitric oxide
produced by the blood sample over a 30 minute time period. FIGS. 1,
3, and 5 show that exposing the blood sample to ABDNAZ under
anaerobic conditions (i.e., N.sub.2 atmosphere) resulted in a
significant increase in the amount of nitric oxide produced by the
blood sample, compared to the amount of nitric oxide produced (i)
without ABDNAZ or (i) with ABDNAZ under aerobic conditions (i.e.,
air atmosphere). FIGS. 6-8 show the amount of nitric oxide formed
in each three-minute period following the start of the
experiment.
Example 2
[0659] The ability of ABDNAZ to enhance blood transfusion during
resuscitation from hemorrhagic shock was evaluated. Experimental
procedures and results are provided below.
Experimental Procedures:
Animal Preparation
[0660] Experiments were performed in 55-65 g male Golden Syrian
Hamsters (Charles River Laboratories, Boston, Mass.) fitted with a
dorsal skinfold window chamber. Animal handling and care followed
the NIH Guide for the Care and Use of Laboratory Animals. The
hamster window chamber model is widely used for microvascular
studies in the unanesthetized state. The complete surgical
technique is described in detail elsewhere, such as in Colantuoni
et al. in Am J Physiol 1984; 246:H508-17. Three to four days after
the initial surgery, the microvasculature was examined and only
animals passing an established systemic and microcirculatory
inclusion criteria, as previously described (e.g., in Cabrales P.
Low dose nitrite enhances perfusion after fluid resuscitation from
hemorrhagic shock. Resuscitation 2009; 80:1431-6), were entered
into the study.
Acute hemorrhage resuscitation protocol
[0661] Acute hemorrhage was induced by withdrawing 50% of estimated
total blood volume (BV) via the carotid artery catheter within 5
min. Total BV was estimated as 7% of body weight. One hour after
hemorrhage induction, animals received 25% of BV of resuscitation
(200 .mu.l/min) via the jugular vein catheter, implemented with the
volume resuscitation strategy defined by the group name, according
to the scheme described before.
Experimental Groups
[0662] Animals were randomly divided into four experimental groups
based on the resuscitation used, namely: [0663] (1) Blood (group
resuscitated with fresh blood only); [0664] (2) Nitrite (group
resuscitated with fresh blood followed by nitrite infusion); [0665]
(3) RRx-001 (group resuscitated with fresh blood treated with
RRx-001 (i.e., ABDNAZ)); [0666] (4) RRx-001+nitrite (group
resuscitated with fresh blood treated with RRx-001 (i.e., ABDNAZ)
followed by nitrite infusion).
[0667] Fresh blood was collected from a donor, adult male Golden
Syrian Hamsters (60-80 g). Briefly, hamster donors were
anaesthetized, left carotid artery catheter was implanted, and
blood was allowed to flow into heparinized tubes (sodium heparin 15
IU/mL). RBCs and plasma were separated by centrifugation (2700 rpm,
7 min). Buffy coat was discarded. RRx-001 treated cells were
prepared by incubation of 1 mL of packed cells with 2 mg of RRx-001
for 30 minutes at 4.degree. C., cells were rinsed 2.times. by
centrifugation at 3,000 rpm with phosphate buffer saline (PBS)
solution with 0.5% albumin (pre-filtered 0.22 .mu.m, pH 7.4). After
the final wash, red blood cells (RBCs) were adjusted to a 30% Hct
with fresh plasma. Although not wishing to be bound by a particular
theory, it is believed that RRx-001 passes through the membrane of
RBCs, binds to, and modifies hemoglobin (Hb).
[0668] For the groups that received "nitrite," 10 .mu.M (in saline
100 .mu.L) of sodium nitrite in saline was infused via the carotid
artery catheter 10 minutes after resuscitation for the groups that
received nitrite. An equal volume of saline was given to the other
groups. To address effects of instrumentation and observation, an
additional Sham group was included.
Experimental Protocol
[0669] Conscious animal was placed in a restraining tube with a
longitudinal slit from which the window chamber protruded, then
fixed to the microscopic stage for transillumination with the
intravital microscope (BX51WI, Olympus, New Hyde Park, N.Y.).
Animals were given 20 min to adjust to the tube environment before
any measurements were made. The tissue image was projected onto a
charge-coupled device camera (4815, COHU, San Diego, Calif.)
connected to a videocassette recorder and viewed on a monitor.
Measurements were carried out using a 40.times. (LUMPFL-WIR,
numerical aperture 0.8, Olympus) water immersion objective.
Systemic (MAP, HR, Hct, Hb, PaO.sub.2, PaCO.sub.2, pH, lactate,
plasma nitrite, and metHb) and microvascular (arteiorlar and
venular diameters, blood flow, and FCD) parameters were analyzed,
as previously described (e.g., in P. Nachuraju et al. in
Resuscitation 2011; 82:607-613; Cabrales P. in Resuscitation 2009;
80:1431-6; Cabrales et al. in Shock 2007; 27:380-9; and Cabrales et
al. in Am J Physiol 2004; 287:H363-73) before hemorrhage
(baseline), after hemorrhage (shock), and up to 90 min after volume
replacement (resuscitation). Tissue viability was measured at 8
hours following hemorrhage as described previously (e.g., in Yang
et al. In Invest Ophthalmol Vis Sci 2003; 44:1993-1997; and
Cabrales et al. in Antioxid Redox Signal 2007; 9:375-84).
Data Analysis
[0670] Table and figure results are presented as mean.+-.SD. Data
within each group were analyzed using analysis of variance for
repeated measurements (Two-way ANOVA). When appropriate, post hoc
analyses were performed with Bonferroni post-test. MetHb and Tissue
viability data were analyzed using the Mann-Whitney U test.
Microhemodynamic data are presented as absolute values and ratios
relative to baseline values. The same vessels and capillary fields
were followed so that direct comparisons to their baseline levels
could be performed, allowing for more robust statistics. All
statistics were calculated using GraphPad Prism 4.03 (GraphPad
Software, Inc., San Diego, Calif.). Changes were considered
statistically significant if P<0.05.
Results:
Systemic Response to Hemorrhage Resuscitation
[0671] Systemic hemodynamic and blood parameters are presented in
Tables 1 and 2. The gold standard for treatment of hemorrhagic
shock is resuscitation via blood transfusion. Thus, with the
exception of blood pressure, treatment effects using RRx-001,
nitrite, or both are compared to the blood only treatment
group.
TABLE-US-00007 TABLE 1 SYSTEMIC PARAMETERS. Hct Hb Nitrite BP HR
(%) (g/dl) (nM) (mmHg) (bpm) Baseline Sham 101 .+-. 5.6 406 .+-. 26
RBCs 50.0 .+-. 0.8 15.2 .+-. 0.2 440 .+-. 32 105.3 .+-. 6.4 416
.+-. 31 Nitrite 49.8 .+-. 0.8 15.2 .+-. 0.3 457 .+-. 23 104.3 .+-.
9.0 428 .+-. 13 RRx-001 49.7 .+-. 1.0 15.1 .+-. 0.3 458 .+-. 21
106.3 .+-. 8.7 409 .+-. 21 RRx-001 + Nitrite 49.1 .+-. 0.9 15.0
.+-. 0.3 439 .+-. 13 107 .+-. 7.4 429 .+-. 22 Shock (50 min) Sham
101.9 .+-. 5.8 412 .+-. 35 RBCs 28.8 .+-. 0.6 8.8 .+-. 0.2 45.5
.+-. 4.8 425 .+-. 43 Nitrite 29.2 .+-. 0.5 8.9 .+-. 0.1 46.1 .+-.
3.6 455 .+-. 21 RRx-001 29.0 .+-. 0.8 8.9 .+-. 0.3 46.3 .+-. 4.0
422.5 .+-. 25 RRx-001 + Nitrite 28.8 .+-. 0.4 8.8 .+-. 0.1 47.8
.+-. 4.4 453 .+-. 31 Resuscitation (30 min) Sham 102.5 .+-. 5.3 426
.+-. 42 RBCs 102.4 .+-. 7.9 359 .+-. 36 Nitrite 91.3 .+-. 6.9 438
.+-. 21 RRx-001 92.5 .+-. 6.7 436 .+-. 12 RRx-001 + 88.7 .+-. 7.4
524 .+-. 44 Nitrite Resuscitation (60 min) Sham 101.5 .+-. 5.2 425
.+-. 44 RBCs 31.4 .+-. 0.4 9.5 .+-. 0.2 79.8 .+-. 6.3 373 .+-. 37
Nitrite 31.0 .+-. 0.7 9.5 .+-. 0.2 96.3 .+-. 7.5 447 .+-. 21
RRx-001 31.1 .+-. 0.6 9.5 .+-. 0.2 98.5 .+-. 7.6 444 .+-. 10
RRx-001 + 31.1 .+-. 0.7 9.4 .+-. 0.2 91.2 .+-. 9.1 517 .+-. 39
Nitrite Resuscitation (90 min) Sham 101.6 .+-. 5.4 426 .+-. 43 RBCs
31.0 .+-. 0.4 9.5 .+-. 0.1 678 .+-. 39 106.5 .+-. 5.7 384 .+-. 47
Nitrite 31.5 .+-. 0.4 9.6 .+-. 0.1 791 .+-. 100 94.7 .+-. 6.5 459
.+-. 14 RRx-001 31.2 .+-. 0.6 9.5 .+-. 0.1 636 .+-. 54 106.3 .+-.
9.0 453 .+-. 17 RRx-001 + 30.6 .+-. 0.4 9.3 .+-. 0.1 799 .+-. 56
99.1 .+-. 1.04 530 .+-. 43 Nitrite
TABLE-US-00008 TABLE 2 ADDITIONAL SYSTEMIC PARAMETERS pO2 pCO2
Lactate pH (mmHg) (mmHg) (mmol/L) Baseline Sham RBCs 7.33 .+-. 0.02
58.9 .+-. 1.5 53.3 .+-. 1.1 1.42 .+-. 0.19 Nitrite 7.34 .+-. 0.02
60.5 .+-. 1.0 52.9 .+-. 1.2 1.30 .+-. 0.13 RRx-001 7.32 .+-. 0.01
59.9 .+-. 2.5 54.2 .+-. 1.8 1.39 .+-. 0.20 RRx-001 + Nitrite 7.33
.+-. 0.02 61.1 .+-. 1.6 53.7 .+-. 1.6 1.37 .+-. 0.19 Shock (50 min)
Sham RBCs 7.29 .+-. 0.01 91.6 .+-. 3.8 37.9 .+-. 0.7 4.37 .+-. 0.30
Nitrite 7.30 .+-. 0.01 93.1 .+-. 2.8 38.9 .+-. 1.3 4.52 .+-. 0.58
RRx-001 7.28 .+-. 0.02 94.9 .+-. 5.1 39.3 .+-. 1.2 4.16 .+-. 0.39
RRx-001 + Nitrite 7.28 .+-. 0.02 98.0 .+-. 3.9 38.4 .+-. 1.2 4.19
.+-. 0.57 Resuscitation (30 min) Sham RBCs Nitrite RRx-001 RRx-001
+ Nitrite Resuscitation (60 min) Sham RBCs 7.34 .+-. 0.01 69.8 .+-.
1.7 50.6 .+-. 1.5 2.08 .+-. 0.22 Nitrite 7.38 .+-. 0.02 76.0 .+-.
2.9 48.3 .+-. 2.2 1.95 .+-. 0.11 RRx-001 7.39 .+-. 0.01 70.9 .+-.
3.4 48.0 .+-. 3.1 1.93 .+-. 0.23 RRx-001 + Nitrite 7.39 .+-. 0.02
79.5 .+-. 4.3 44.7 .+-. 2.1 1.41 .+-. 0.12 Resuscitation (90 min)
Sham RBCs 7.35 .+-. 0.02 63.1 .+-. 1.6 51.0 .+-. 1.8 1.86 .+-. 0.18
Nitrite 7.36 .+-. 0.02 65.2 .+-. 3.0 45.0 .+-. 2.4 1.71 .+-. 0.20
RRx-001 7.36 .+-. 0.01 68.7 .+-. 2.9 49.0 .+-. 2.2 1.63 .+-. 0.23
RRx-001 + Nitrite 7.38 .+-. 0.02 70.4 .+-. 3.9 50.7 .+-. 2.7 1.41
.+-. 0.14
[0672] During hemorrhagic shock, Hct and Hb dropped about 50% for
all treatment groups and no significant differences were seen
between groups at any time during the study. Although resuscitation
partially restored blood pressure (MAP) from shock in all treatment
groups, MAP was significantly decreased in the RRx-001 group at 60
minutes. Compared to the blood group, significant differences in
MAP and heart rate were seen in the blood+nitrite, RRx-001, and
RRx-001+nitrite groups following resuscitation. For the nitrite
group, MAP was significantly decreased at 30 and 90 minutes. MAP in
the RRx-001, was only decreased at 30 minutes and in the
RRx-001+nitrite group it was significantly decreased at 30 and 60
minutes. HR for the nitrite, RRx-001, and RRx-001+nitrite treatment
groups compared to the blood group was significantly increased.
[0673] Hemorrhagic shock decreased arterial pH and pCO2,
significantly compromising acid based balance relative to baseline.
Resuscitation partially recovered all blood gas parameters. In the
nitrite, RRx-001, and RRx-001+nitrite groups, pH was significantly
increased at 60 minutes post resuscitation compared to the blood
group. For the RRx-001+nitrite group, pH was also significantly
increased at 90 minutes post resuscitation. pO2 levels were
significantly increased: in the in the nitrite group at 60 minutes
post resuscitation; in the RRx-001 group at 90 minutes post
resuscitation; in the RRx-001+nitrite group, during 60 minutes of
shock and at 60 and 90 minutes post resuscitation. Compared to the
blood group, pCO2 levels in the RRx-001 group were not
significantly different. In the RRx-001+nitrite group, lactate
(lact) levels were significantly decreased at 60 and 90 minutes
post-resuscitation. Decreases in lactate levels in the nitrite and
RRx-001 groups did not reach significance.
[0674] The percent MetHb levels for the nitrite, RRx-001, and
RRx-001+nitrite groups are presented in FIG. 9. In normal, healthy
individuals metHb levels are about 1% of the total Hb and
methemoglobinemia occurs when the concentration of metHb in the
blood exceeds 1.5 g/dL (8%-12% of the normal Hb level). See, for
example, Hamirani et al. in Tex Heart Inst J 2008; 35:76-77. Of the
three treatment groups, only RRx-001 had normal metHb levels.
Microvascular Measurements
[0675] Changes in arteriolar diameter and blood flow during the
hemorrhagic shock resuscitation protocol for all experimental
groups are presented in FIG. 10. Compared to the blood group,
arterial diameter and blood flow were increased in all treatment
groups. However, these increases only reached significance in the
RRx-001 and RRx-001+nitrite groups. Arteriolar diameter was
significantly increased at 60 and 90 minutes (P<0.01 and
P<0.05, respectively) only in RRx-001+nitrite following
resuscitation and blood flow was significantly (P<0.001)
increased in the both the RRx-001 and RRx-001+nitrite groups at 60
and 90 minutes. At 30 minutes post resuscitation, the difference in
blood flow between the blood only and RRx-001+nitrite groups was
also significant (P<0.05).
[0676] Changes in the number of capillaries perfused with RBCs
during the protocol are presented in FIG. 11. Resuscitation
partially restored hemorrhage induced reductions in functional
capillary density (FCD) in all groups. Again, compared to the blood
group, FCD was not significantly different in the nitrite group
following resuscitation. However, both RRx-001 and RRx-001+nitrite
treatment resulted in significant (P<0.05 and P<0.01,
respectively) increases in FCD at 90 minutes post resuscitation.
Statistical significance (P<0.05) was also observed at 60
minutes in the RRx-001+nitrite group.
[0677] Calculated vascular resistance (average MAP/average blood
flow) relative to baseline is presented in FIG. 12. Peripheral
vascular resistance for all groups increased after hemorrhagic
shock (about 1.5 times the resistance at baseline) and decreased
after resuscitation. The RRx-001 and RRx-001+nitrite groups had a
similar impact on vascular resistance.
Tissue Viability
[0678] Tissue viability (the number of apoptotic and necrotic cells
in 40 fields) for all treatment groups at 8 hours following
resuscitation is presented in FIG. 13. The number of apoptotic
cells in the RRx-001 and RRx-001+nitrite groups was significantly
(P<0.01) less than the blood group. The number of necrotic cells
was also significantly (P<0.01) less in the RRx-001 and
RRx-001+nitrite groups compared to the blood group. Supplementation
with nitrite also significantly reduced the number of necrotic
cells.
DISCUSSION
[0679] Experimental results show that RRx-001 treated blood with or
without nitrite supplementation provides superior systemic and
microvascular hemodynamic responses compared to blood transfusion
with or without nitrite. Incorporating RRx-001 into
transfusion-based resuscitation affords the added benefit of
selectively increasing NO generation under hypoxic conditions.
Without being bound by a particular theory, it is believed that
RRx-001 generates NO in two ways: i) as an NO donor: through
metabolism of the dinitro groups released from the compound, and
ii) as an NO promoter: beta-Cys-93 modification by RRx-001 enhances
hypoxia-mediated nitrite reduction to NO by deoxyhemoglobin. Our
results demonstrate that these RRx-001 mediated benefits improved
systemic and microvascular parameters, which appears to correlate
with tissue viability. Thus, RRx-001 treated blood should minimize
short and long term organ damage after hemorrhagic shock.
[0680] Hemorrhagic hypotension leads to a well-characterized
sequence of events, and ultimately to vascular decompensation, due
to a continuous increase in peripheral vascular resistance. The
outcome of hemorrhagic shock is related to the degree of
hypovolemia, the magnitude of acquired oxygen debt, and the delay
in treatment. Monitoring the microcirculation is crucial in
determining the effect of changes in intravascular volume in tissue
hypo-perfusion. Application of various techniques, including
intravital microscopy, has shown the presence of major
microcirculatory alterations during hemorrhage, and the persistence
of these microcirculatory alterations have been associated with
multiorgan failure and death. See, for example, Sinaasappel et al.
in J Physiol 1999; 514(Pt 1):245-253; and Ellis et al. in Crit Care
2005; 9(Suppl 4):53-8.
[0681] Blood transfusion is currently the gold standard for
treatment of severe hemorrhagic shock. When blood is used during
resuscitation, intravascular blood volume and oxygen carrying
capacity are restored, cardiovascular function improves, energy
requirements are met, and survival more likely. Practically
however, transfusion post hemorrhage recovers the microcirculation,
but not necessarily to normal levels. The injury resulting from the
shock phase prior to resuscitation limits perfusion during the
resuscitation and thus prevents full recovery of the
microcirculation immediately post resuscitation. Moreover, when
blood is used during resuscitation, "normal" MAP is restored,
however restoring MAP is not necessarily accompanied by the
restoration of organ perfusion and oxygenation, due to
microvascular flow dysfunctions (the so-called "no reflow"
phenomenon). See, for example, Zakaria et al. in J Trauma 2005;
58:499-508; and Rezkalla et al. in Circulation 2002; 105:656-62.
During the shock phase and immediately post resuscitation, vascular
endothelial shear stress and endothelium NO synthase (eNOS)
activity is also impaired and results in delayed dilation of the
endothelium. Over time, eNOS activity and microvascular flow
dysfunction recover. However, if the resuscitation is inadequate
during this critical period multi-organ injury can ensue. The
results of our study suggest that during the time when NO synthase
is still malfunctioning, incorporation of RRx-001 with blood
resuscitation, via restoration of intravascular NO concentration,
would increase perfusion by relaxing arterioles and lowering
vascular resistance leading to improved microvascular function,
reduced cell death, and preserved tissue viability, ensuring a
better overall outcome compared to blood transfusion alone.
[0682] The use of NO donors under conditions of hemorrhagic shock
have been shown to result in enhanced myocardial contractile
activity that leads to a situation where mean arterial pressure
does not decrease further despite significant decrease of total
peripheral resistance. Remizova and colleagues studied the effects
of an NO donor, DNIC-GS (dinitrosyl iron complexes with
glutathione) in a hemorrhagic shock model. See, for example,
Remizova et al. in Eur J Pharmacol 2011; 662:40-46. They found that
injection of DNIC-GS into the blood flow of rats prior to
hemorrhage by increased stroke volume, left ventricular work, and
cardiac output. The results of our study indicate that RRx-001
should improve these indices of cardiac function in the face of
decreased vascular resistance.
[0683] Nitrite, a biologic metabolite of NO, present in a variety
of foods. Nitrite has been appreciated as an inflammatory mediator
of nitration reactions and a precursor for NO under acidic or
ischemic conditions and plasma nitrite levels correlate with eNOS
activity and are tightly controlled. We have previously studied the
effects of nitrite supplementation (10 .mu.M and 50 .mu.M nitrite)
on systemic (BP, HR, pH, pO2, pCO2) and microvascular parameters
(arteriolar diameter, blood flow, FCD) after resuscitation from
hemorrhagic shock. Similar effects on systemic and microvascular
parameters were observed with the administration of 10 .mu.M
nitrate compared to the nitrite group in the present study. By
comparison, administration of 50 .mu.M nitrite had a more profound
effect on arteriolar diameter and blood flow but negatively
affected blood pressure and metHb levels. Blood pressure in the 50
.mu.M nitrite group was significantly decreased compared to 0 .mu.M
group (control group) at 60 minutes following resuscitation and at
60 and 90 minutes; % metHb was 5.8.+-.1.8 and 3.1.+-.1.3,
respectively. In the current study, RRx-001 treatment maintained
blood pressure following resuscitation and resulted in metHb levels
of only 1.4.+-.0.1 at 60 minutes and 1.2.+-.0.1 at 90 minutes which
corresponds to metHb levels in healthy individuals of about 1% of
the total Hb: Methemoglobinemia occurs when the concentration of
metHb in the blood exceeds 1.5 g/dL (8%-12% of the normal Hb
level), where tissue oxygenation is compromised.
INCORPORATION BY REFERENCE
[0684] The entire disclosure of each of the patent documents and
scientific articles referred to herein is incorporated by reference
for all purposes.
EQUIVALENTS
[0685] The invention may be embodied in other specific forms
without departing from the spirit or essential characteristics
thereof. The foregoing embodiments are therefore to be considered
in all respects illustrative rather than limiting the invention
described herein. Scope of the invention is thus indicated by the
appended claims rather than by the foregoing description, and all
changes that come within the meaning and range of equivalency of
the claims are intended to be embraced therein.
* * * * *