U.S. patent application number 14/728592 was filed with the patent office on 2016-04-28 for methods and compositions for treating viral or virally-induced conditions.
The applicant listed for this patent is HEMAQUEST PHARMACEUTICALS, INC., TRUSTEES OF BOSTON UNIVERSITY. Invention is credited to Ronald J. Berenson, Douglas V. Faller.
Application Number | 20160113929 14/728592 |
Document ID | / |
Family ID | 44564167 |
Filed Date | 2016-04-28 |
United States Patent
Application |
20160113929 |
Kind Code |
A1 |
Berenson; Ronald J. ; et
al. |
April 28, 2016 |
METHODS AND COMPOSITIONS FOR TREATING VIRAL OR VIRALLY-INDUCED
CONDITIONS
Abstract
Provided are methods and compositions for the prevention and/or
treatment of viral conditions, virally-induced conditions and
inflammatory conditions. The methods can comprise administering to
a subject a viral inducing agent with an antiviral agent, and
optionally an additional agent. The viral inducing agent can be a
HDAC inhibitor administered orally.
Inventors: |
Berenson; Ronald J.; (Mercer
Island, WA) ; Faller; Douglas V.; (Weston,
MA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
HEMAQUEST PHARMACEUTICALS, INC.
TRUSTEES OF BOSTON UNIVERSITY |
San Diego
Boston |
CA
MA |
US
US |
|
|
Family ID: |
44564167 |
Appl. No.: |
14/728592 |
Filed: |
June 2, 2015 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
13912637 |
Jun 7, 2013 |
|
|
|
14728592 |
|
|
|
|
13046555 |
Mar 11, 2011 |
|
|
|
13912637 |
|
|
|
|
61430931 |
Jan 7, 2011 |
|
|
|
61313052 |
Mar 11, 2010 |
|
|
|
Current U.S.
Class: |
514/3.7 ;
514/263.31; 514/263.32 |
Current CPC
Class: |
A61P 31/12 20180101;
A61K 31/19 20130101; A61K 45/06 20130101; A61K 31/27 20130101; A61K
31/4402 20130101; A61P 29/00 20180101; A61K 9/0053 20130101; A61K
31/506 20130101; Y02A 50/467 20180101; A61K 31/18 20130101; A61K
31/165 20130101; A61K 31/4045 20130101; A61P 31/20 20180101; A61K
31/167 20130101; A61K 38/12 20130101; Y02A 50/30 20180101; A61K
31/473 20130101; A61K 31/522 20130101; A61P 31/22 20180101; A61K
38/15 20130101; A61K 31/513 20130101; A61K 31/52 20130101; A61K
31/167 20130101; A61K 2300/00 20130101; A61K 31/19 20130101; A61K
2300/00 20130101; A61K 31/27 20130101; A61K 2300/00 20130101; A61K
31/4045 20130101; A61K 2300/00 20130101; A61K 31/506 20130101; A61K
2300/00 20130101; A61K 31/513 20130101; A61K 2300/00 20130101; A61K
38/12 20130101; A61K 2300/00 20130101 |
International
Class: |
A61K 31/506 20060101
A61K031/506; A61K 31/522 20060101 A61K031/522; A61K 31/19 20060101
A61K031/19; A61K 38/12 20060101 A61K038/12; A61K 31/167 20060101
A61K031/167; A61K 31/4045 20060101 A61K031/4045; A61K 31/165
20060101 A61K031/165; A61K 9/00 20060101 A61K009/00; A61K 31/473
20060101 A61K031/473 |
Claims
1.-139. (canceled)
140. A method for treating and/or preventing a viral or
virally-induced condition comprising administering a HDAC inhibitor
and an antiviral agent wherein the viral or virally-induced
condition is caused by a DNA virus and the HDAC inhibitor is
administered at dose of less than 2 mg/kg per dose.
141. The method of claim 140, wherein the HDAC inhibitor is
CHR-3996.
142. The method of claim 140, wherein the DNA virus is a
herpesvirus.
143. The method of claim 140, wherein the DNA virus is an
Epstein-Barr virus.
144. The method of claim 140, wherein the DNA virus is a
cytomegalovirus.
145. The method of claim 140, wherein the DNA virus is a varicella
zoster virus.
146. The method of claim 140, wherein the virally-induced condition
is a lymphoma, chronic lymphocytic leukemia, nasopharyngeal
carcinoma, gastric cancer, Kaposi's sarcoma, rheumatoid arthritis,
systemic lupus erythematosus, or multiple sclerosis.
147. The method of claim 140, wherein the antiviral agent is
acyclovir (ACV), ganciclovir (GCV), valganciclovir, famciclovir,
foscarnet, ribavirin, zalcitabine (ddC), zidovudine (AZT),
stavudine (D4T), lamivudine (3TC), didanosine (ddI), cytarabine,
dideoxyadenosine, edoxudine, floxuridine, idozuridine, inosine
pranobex, 2'-deoxy-5-(methylamino)uridine, trifluridine and
vidarabine.
148. The method of claim 140, wherein the antiviral agent is
ganciclovir (GCV).
149. A method for treating and/or preventing a viral or
virally-induced condition comprising administering a HDAC inhibitor
and an antiviral agent wherein the viral or virally-induced
condition is caused by a retrovirus and the HDAC inhibitor is
administered at dose of less than 2 mg/kg per dose.
150. The method of claim 149, wherein the HDAC inhibitor is
CHR-3996.
151. The method of claim 149, wherein the retrovirus is HIV, HTLV1
or HTLV2.
152. The method of claim 149, wherein the retrovirus is HIV.
153. A composition comprising a (i) HDAC inhibitor and (ii) an
antiviral agent.
154. The composition of claim 153, wherein the HDAC inhibitor is
CHR-3996.
155. The composition of claim 153, wherein the antiviral a
herpesvirus antiviral.
156. The composition of claim 153, wherein the antiviral agent is
an anti-HIV antiviral.
157. The composition of claim 153, wherein the antiviral agent is
acyclovir (ACV), ganciclovir (GCV), valganciclovir, famciclovir,
foscarnet, ribavirin, zalcitabine (ddC), zidovudine (AZT),
stavudine (D4T), lamivudine (3TC), didanosine (ddI), cytarabine,
dideoxyadenosine, edoxudine, floxuridine, idozuridine, inosine
pranobex, 2'-deoxy-5-(methylamino)uridine, trifluridine and
vidarabine.
158. The composition of claim 153, wherein the antiviral agent is
ganciclovir (GCV).
159. The composition of claim 153, formulated for oral
administration.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation application of co-pending
U.S. patent application Ser. No. 13/912,637 filed Jun. 7, 2013,
which is a continuation application of U.S. patent application Ser.
No. 13/046,555 filed on Mar. 11, 2011, which claims the benefit
under 35 U.S.C. .sctn.119(e) of U.S. Provisional Application No.
61/430,931 filed on Jan. 7, 2011, and U.S. Provisional Application
No. 61/313,052 filed on Mar. 11, 2010, the contents of each of
which are incorporated herein by reference in their entireties.
SEQUENCE LISTING
[0002] The instant application contains a Sequence Listing which
has been submitted in ASCII format via EFS-Web and is hereby
incorporated by reference in its entirety. Said ASCII copy, created
on Jun. 1, 2015, is named 701586-070083_SL.txt and is 1,296 bytes
in size.
BACKGROUND OF THE INVENTION
[0003] Many patients can have latent infections in which a virus is
present but is not expressing viral proteins such as viral
thymidine kinase or protein kinase, the target for common
anti-viral drugs such as acyclovir and ganciclovir. A viral
inducing drug such as a histone deacetylase inhibitor (HDAC
inhibitor--HDACi) can be used to re-induce the expression of viral
thymidine kinase or protein kinase in viral infected cells in the
subject; the subject can then be treatment with antiviral drugs to
eliminate latent viral infections. As EBV and/or other latent viral
infections can be associated with a variety of conditions, many of
which are inflammatory conditions, such as lymphomas, autoimmune
conditions, allergic conditions, eliminating the latent virus with
this therapy can be used to prevent or treat such conditions.
[0004] EBV can induce autoimmune conditions through a number of
potential mechanisms, e.g.: 1) activating B cells to produce
auto-antibodies, 2) turning on T cells that attack host tissue, 3)
molecular mimicry in which EBV antigens cross react with host
antigens such that autoimmune condition results when T cells or
antibodies reactive with these antigens cross react with host
antigens causing damage to host tissues, 4) EBV infected B cells
produce cytokines which turn on other elements of the immune system
and increase inflammation which can also exacerbate autoimmune
condition, 5) EBV infected B or T cells can become immortalized via
EBV proteins indirectly turning out anti-apoptotic or survival
pathways. Autoimmune conditions with evidence of an EBV
relationship include multiple sclerosis, systemic lupus
erythematosus, rheumatoid arthritis and Sjogren's syndrome.
Cytomegalovirus and herpes simplex virus have been associated with
coronary artery condition.
[0005] There is a need for methods of treating and/or preventing
viral conditions, viral-induced conditions, and related
inflammatory conditions.
SUMMARY OF THE INVENTION
[0006] In one aspect, a method for treating and/or preventing a
viral condition, a virally-induced condition, or an inflammatory
condition comprising administering a histone deacetylase inhibitor
(HDAC inhibitor) and an antiviral agent wherein the HDAC inhibitor
is a pyrimidine hydroxamic acid derivative is provided.
[0007] In another aspect, a method for treating and/or preventing a
viral or virally-induced condition is provided comprising
administering a HDAC inhibitor and an antiviral agent wherein the
viral or virally-induced condition is caused by a DNA virus and the
HDAC inhibitor is administered at dose of less than 2 mg/kg per
dose.
[0008] In yet another aspect, a method for treating and/or
preventing a viral condition, a virally-induced condition, or an
inflammatory condition is provided comprising administering a HDAC
inhibitor and an antiviral agent wherein the MW of the HDAC
inhibitor is greater than 275 g/mol.
[0009] In another aspect, a method for treating and/or preventing a
viral condition or a virally-induced condition is provided,
comprising administering a HDAC inhibitor wherein the HDAC
inhibitor is a pyrimidine hydroxamic acid derivative.
[0010] In another aspect, a composition comprising a (i) HDAC
inhibitor and (ii) an antiviral agent is provided, wherein the HDAC
inhibitor is a pyrimidine hydroxamic acid derivative.
[0011] Provided herein, in one aspect, is a method for treating
and/or preventing a viral or virally-induced condition comprising
administering a HDAC inhibitor and an antiviral agent wherein the
viral or virally-induced condition is caused by a DNA virus and the
HDAC inhibitor is administered at dose of less than 2 mg/kg per
dose. In some embodiments, the HDAC inhibitor is
Vorinostat/suberoyl anilide hydroxamic acid, JNJ-26481585
(N-hydroxy-2-(4-((((1-methyl-1H-indol-3-yl)methyl)amino)methyl)piperidin--
1-yl)pyrimidine-5-carboxamide), R306465/JNJ-16241199
(N-hydroxy-5-(4-(naphthalen-2-ylsulfonyl)piperazin-1-yl)pyrimidine-2-carb-
oxamide), CHR-3996
(2-(6-{[(6-Fluoroquinolin-2-yl)methyl]amino}-3-azabicyclo[3.1.0]hex-3-yl)-
-N-hydroxypyrimidine-5-carboxamide), Belinostat/PXD101,
Panobinostat/LBH-589, trichostatin A/TSA
(7-[4-(dimethylamino)phenyl]-N-hydroxy-4,6-dimethyl-7-oxohepta-2,4-dienam-
ide), ITF2357, CBHA, Givinostat/ITF2357, romidepsin (Istodax.TM.),
PCI-24781, depsipeptide (FR901228 or FK228), butyrate,
phenylbutyrate, valproic acid, AN-9, CI-994, Entinostat/MS-275,
SNDX-275, mocetinostat/MGCD0103
(N-(2-aminophenyl)-4-((4-pyridin-3-ylpyrimidin-2-ylamino)methyl)benzamide-
), m-carboxycinnamic acid, bishydroxamic acid, suberic
bishydroxamic acid, oxamflatin, ABHA, SB-55629, pyroxamide,
propenamides, aroyl pyrrolyl hydroxamides, and LAQ824
(((E)-N-hydroxy-3-[4-[[2-hydroxyethyl-[2-(1H-indol-3-yl)ethyl]amino]methy-
l]phenyl]prop-2-enamide). In other embodiments, the HDAC inhibitor
is a largazole derivative. In certain embodiments, the HDAC
inhibitor is a N-hydroxypyrimidine-5-carboxamide, wherein the HDAC
inhibitor comprises an azabicyclo-hexane, wherein the HDAC
inhibitor comprises a fluoroquinoline group, or wherein the HDAC
inhibitor is a non-piperidine-containing pyrimidine hydroxamic acid
derivative.
[0012] In certain embodiments, the DNA virus is a herpes virus. In
some embodiments, the herpes virus is an Epstein-Barr virus.
[0013] In some embodiments, the HDAC inhibitor and the antiviral
agent are co-formulated. In certain embodiments, the co-formulation
comprises a unit dose of no greater than 80 mg of the HDAC
inhibitor and no greater than 1500 mg of the antiviral agent.
[0014] In certain embodiments, the HDAC inhibitor can penetrate the
blood brain barrier.
[0015] In some embodiments, the method further comprises
administering an additional agent. In certain embodiments, the
additional agent is an antiviral agent, a HDAC inhibitor, or a
chemotherapeutic.
[0016] In certain embodiments, the virally-induced condition is a
cancerous condition, an inflammatory condition, an autoimmune
condition, an allergic condition, or a skin condition. In some
embodiments, the virally-induced condition is a lymphoma, chronic
lymphocytic leukemia, nasopharyngeal carcinoma, gastric cancer,
Kaposi's sarcoma, rheumatoid arthritis, systemic lupus
erythematosus, or multiple sclerosis.
[0017] Also provided herein, in an additional aspect, is a method
for treating and/or preventing a viral condition, a virally-induced
condition, or an inflammatory condition comprising administering a
HDAC inhibitor and an antiviral agent wherein the MW of the HDAC
inhibitor is greater than 275 g/mol. In some embodiments, the HDAC
inhibitor is a N-hydroxypyrimidine-5-carboxamide, wherein the HDAC
inhibitor comprises an azabicyclo-hexane, wherein the HDAC
inhibitor comprises a fluoroquinoline group, or wherein the HDAC
inhibitor is a non-piperidine-containing pyrimidine hydroxamic acid
derivative. In certain embodiments, the HDAC inhibitor is
JNJ-26481585
(N-hydroxy-2-(4-((((1-methyl-1H-indol-3-yl)methyl)amino)methyl)piperidin--
1-yl)pyrimidine-5-carboxamide), R306465/JNJ-16241199
(N-hydroxy-5-(4-(naphthalen-2-ylsulfonyl)piperazin-1-yl)pyrimidine-2-carb-
oxamide), or CHR-3996
(2-(6-{[(6-Fluoroquinolin-2-yl)methyl]amino}-3-azabicyclo[3.1.0]hex-3-yl)-
-N-hydroxypyrimidine-5-carboxamide). In other embodiments, the HDAC
inhibitor is a largazole derivative.
[0018] In some embodiments, the viral or virally induced condition
is caused by a herpes virus. In certain embodiments, the herpes
virus is an Epstein-Ban virus.
[0019] In some embodiments, the HDAC inhibitor and the antiviral
agent are administered simultaneously.
[0020] In certain embodiments, the HDAC inhibitor and the antiviral
agent are administered orally.
[0021] In some embodiments, the HDAC inhibitor is administered at
0.01-1 mg/kg per dose. In certain embodiments, 1, 2, 3, or 4 doses
are administered daily. In some embodiments, the total daily dosage
of the HDAC inhibitor is no greater than about 200 mg. In certain
embodiments, the antiviral agent is valganciclovir and is
administered at dose of 500-1500 mg/dose.
[0022] In some embodiments, the HDAC inhibitor and the antiviral
agent are co-formulated. In certain embodiments, the co-formulation
comprises a unit dose of no greater than 80 mg of the HDAC
inhibitor and no greater than 1500 mg of the antiviral agent. In
some embodiments, the antiviral agent is valganciclovir, and the
valganciclovir is a timed release or slow release oral
formulation.
[0023] In certain embodiments, the viral condition or
virally-induced condition is caused by a human immunodeficiency
virus, a herpes virus, a parvovirus, a coxsackie virus, a Human
T-lymphotropic virus, a BK virus, or a hepatitis virus. In some
embodiments, the viral condition or virally-induced condition is
caused by a retrovirus or a herpes virus. In certain embodiments,
the herpes virus is a herpes simplex virus, a herpes genitalis
virus, a varicella zoster virus, an Epstein-Barr virus, a human
herpes virus 6, a herpes virus type 1, herpes virus type 2, a human
herpes virus type 8, or a cytomegalovirus. In some embodiments, the
herpes virus is the Epstein-Barr virus. In certain embodiments, the
viral condition or virally-induced condition is caused by a DNA
virus. In some embodiments, the viral condition or virally-induced
condition is not caused by a retrovirus.
[0024] In certain embodiments, the virally-induced condition is a
cancer, an inflammatory condition, an allergic condition, an
autoimmune condition, or a skin condition. In some embodiments, the
virally-induced condition is lymphoma, chronic lymphocytic
leukemia, nasopharyngeal carcinoma, gastric cancer, Kaposi's
sarcoma, rheumatoid arthritis, systemic lupus erythematosus, or
multiple sclerosis. In certain embodiments, the virally-induced
condition is not sepsis or viremia. In some embodiments, the
inflammatory condition is an autoimmune condition, and allergic
condition, or a skin condition.
[0025] In certain embodiments, the HDAC inhibitor can penetrate the
blood brain barrier. In some embodiments, the antiviral agent is a
HIV drug, a Herpes drug, a CMV drug, or a hepatitis drug. In
certain embodiments, the antiviral agent is acyclovir, ganciclovir
or valganciclovir. In some embodiments, the antiviral agent is not
a heat shock protein inhibitor, an immunosuppressant, an
antibiotic, a glucocorticoid, a non-steroidal anti-inflammatory
drug, a Cox-2-specific inhibitor or a TNF-.alpha. binding protein.
In certain embodiments, the antiviral agent is not a Hsp90
inhibitor, tacrolimus, cyclosporin, rapamycin (sirolimus),
methotrexate, cyclophosphamide, azathioprine, mercaptopurine,
mycophenolate, FTY720, levofloxacin, amoxycillin, prednisone,
cortisone acetate, prednisolone, methylprednisolone, dexamethasone,
betamethasone, triamcinolone, beclometasone, fludrocortisone
acetate, deoxycorticosterone acetate, aldosterone, salicylates,
arylalkanoic acids, a 2-arylpropionic acid, a N-arylanthranilic
acid, an oxicam, a coxib, a sulphonanilide, valdecoxib, celecoxib,
rofecoxib, leflunomide, gold thioglucose, gold thiomalate, aurofin,
sulfasalazine, hydroxychloroquinine, minocycline, infliximab,
etanercept, adalimumab, abatacept, anakinra, interferon-.beta.,
interferon-.gamma., interleukin-2, an allergy vaccine, an
antihistamine, an antileukotriene, a beta-agonist, theophylline, or
an anticholinergic.
[0026] In some embodiments, the method further comprises
administering an additional agent. In certain embodiments, the
additional agent is an antiviral agent, a HDAC inhibitor, or a
chemotherapeutic.
[0027] Further provided herein, in one aspect, is a method for
treating and/or preventing a viral condition, a virally-induced
condition, or an inflammatory condition comprising administering a
HDAC inhibitor and an antiviral agent, wherein the HDAC inhibitor
is a heterocyclic hydroxamic acid derivative. In some embodiments,
the HDAC inhibitor is a pyrimidine hydroxamic acid derivative. In
certain embodiments, the HDAC inhibitor is a
N-hydroxypyrimidine-5-carboxamide, wherein the HDAC inhibitor
comprises an azabicyclo-hexane, wherein the HDAC inhibitor
comprises a fluoroquinoline group, or wherein the HDAC inhibitor
does not comprise a piperidine group. In some embodiments, the HDAC
inhibitor is JNJ-26481585
(N-hydroxy-2-(4-((((1-methyl-1H-indol-3-yl)methyl)amino)methyl)piperidin--
1-yl)pyrimidine-5-carboxamide), R306465/JNJ-16241199
(N-hydroxy-5-(4-(naphthalen-2-ylsulfonyl)piperazin-1-yl)pyrimidine-2-carb-
oxamide), or CHR-3996
(2-(6-{[(6-Fluoroquinolin-2-yl)methyl]amino}-3-azabicyclo[3.1.0]hex-3-yl)-
-N-hydroxypyrimidine-5-carboxamide).
[0028] In some embodiments, the HDAC inhibitor and the antiviral
agent are administered simultaneously. In certain embodiments, the
HDAC inhibitor and the antiviral agent are administered orally. In
some embodiments, the antiviral agent is valganciclovir. In certain
embodiments, the HDAC inhibitor is administered at 0.2-2 mg/kg per
dose. In some embodiments, 1, 2, 3, or 4 doses are administered
daily. In certain embodiments, the total daily dosage of the HDAC
inhibitor is no greater than about 100 mg. In some embodiments,
valganciclovir is administered at dose of 900 mg/dose or less.
[0029] In certain embodiments, the HDAC inhibitor and the antiviral
agent are co-formulated. In some embodiments, the co-formulation
comprises a unit dose of no greater than 80 mg of the HDAC
inhibitor and no greater than 1500 mg of the antiviral agent.
[0030] In certain embodiments, the viral condition or
virally-induced condition is caused by a human immunodeficiency
virus, a herpes virus, a parvovirus, a coxsackie virus, a Human
T-lymphotropic virus, a BK virus, or a hepatitis virus. In specific
embodiments, the herpes virus is a herpes simplex virus, a herpes
genitalis virus, a varicella zoster virus, an Epstein-Barr virus, a
human herpes virus 6, a herpes virus type 1, herpes virus type 2, a
human herpes virus type 8, or a cytomegalovirus.
[0031] In some embodiments, the viral condition or virally-induced
condition is caused by a DNA virus. In certain embodiments, the
viral condition or virally-induced condition is caused by an
Epstein-Barr virus. In some embodiments, the viral condition or
virally-induced condition is not caused by a retrovirus. In certain
embodiments, the virally-induced condition is a cancer, an
inflammatory condition, an allergic condition, an autoimmune
condition, or a skin condition. In some embodiments, the
virally-induced condition is lymphoma, chronic lymphocytic
leukemia, nasopharyngeal carcinoma, gastric cancer, Kaposi's
sarcoma, rheumatoid arthritis, systemic lupus erythematosus, or
multiple sclerosis. In certain embodiments, the virally-induced
condition is not sepsis or viremia. In specific embodiments, the
inflammatory condition is an autoimmune condition, and allergic
condition, or a skin condition.
[0032] In some embodiments, the HDAC inhibitor can penetrate the
blood brain barrier.
[0033] In certain embodiments, the antiviral agent is a HIV drug, a
Herpes drug, a CMV drug, or a hepatitis drug. In some embodiments,
the antiviral agent is acyclovir, ganciclovir, or valganciclovir.
In certain embodiments, the antiviral agent is not a heat shock
protein inhibitor, an immunosuppressant, an antibiotic, a
glucocorticoid, a non-steroidal anti-inflammatory drug, a
Cox-2-specific inhibitor or a TNF-.alpha. binding protein. In other
embodiments, the antiviral agent is not a Hsp90 inhibitor,
tacrolimus, cyclosporin, rapamycin (sirolimus), methotrexate,
cyclophosphamide, azathioprine, mercaptopurine, mycophenolate,
FTY720, levofloxacin, amoxycillin, prednisone, cortisone acetate,
prednisolone, methylprednisolone, dexamethasone, betamethasone,
triamcinolone, beclometasone, fludrocortisone acetate,
deoxycorticosterone acetate, aldosterone, salicylates, arylalkanoic
acids, a 2-arylpropionic acid, a N-arylanthranilic acid, an oxicam,
a coxib, a sulphonanilide, valdecoxib, celecoxib, rofecoxib,
leflunomide, gold thioglucose, gold thiomalate, aurofin,
sulfasalazine, hydroxychloroquinine, minocycline, infliximab,
etanercept, adalimumab, abatacept, anakinra, interferon-.beta.,
interferon-.gamma., interleukin-2, an allergy vaccine, an
antihistamine, an antileukotriene, a beta-agonist, theophylline, or
an anticholinergic.
[0034] In some embodiments, the method further comprises
administering an additional agent. In certain embodiments, the
additional agent is an antiviral agent, a HDAC inhibitor, or a
chemotherapeutic therapy.
[0035] Also provided herein, in another aspect, is a method for
treating and/or preventing a viral condition or a virally-induced
condition comprising administering a HDAC inhibitor wherein the
HDAC inhibitor is a pyrimidine hydroxamic acid derivative
comprising an azabicyclo-hexane, or wherein the HDAC inhibitor is a
non-piperidine-containing pyrimidine hydroxamic acid derivative. In
some embodiments, the HDAC inhibitor is a pyrimidine hydroxamic
acid derivative comprising an azabicyclo-hexane. In certain
embodiments, the HDAC inhibitor is a non-piperidine-containing
pyrimidine hydroxamic acid derivative. In some embodiments, the
HDAC inhibitor comprises a fluoroquinoline group.
[0036] In some embodiments, the method further comprises
administering an antiviral agent. In certain embodiments, the HDAC
inhibitor and antiviral agent are administered simultaneously. In
some embodiments, the HDAC inhibitor is administered orally. In
certain embodiments, the HDAC inhibitor is administered at 0.01-1
mg/kg per dose. In some embodiments, 1, 2, 3, or 4 doses are
administered daily. In certain embodiments, the total daily dosage
of the HDAC inhibitor is no greater than about 200 mg. In some
embodiments, the antiviral agent is administered at dose of
500-1500 mg/dose.
[0037] In certain embodiments, the HDAC inhibitor and the antiviral
agent are co-formulated. In some embodiments, the co-formulation
comprises a unit dose of no greater than 80 mg of the HDAC
inhibitor and no greater than 1500 mg of the antiviral agent. In
certain embodiments, the antiviral agent is valganciclovir.
[0038] In some embodiments, the viral condition or virally-induced
condition is caused by a human immunodeficiency virus, a herpes
virus, a parvovirus, a coxsackie virus, a Human T-lymphotropic
virus, a BK virus, or a hepatitis virus. In specific embodiments,
the herpes virus is a herpes simplex virus, a herpes genitalis
virus, a varicella zoster virus, an Epstein-Barr virus, a human
herpes virus 6, a herpes virus type 1, herpes virus type 2, a human
herpes virus type 8, or a cytomegalovirus. In some embodiments, the
herpes virus is the Epstein-Barr virus.
[0039] In certain embodiments, the viral condition or
virally-induced condition is caused by a DNA virus. In some
embodiments, the viral condition or virally-induced condition is
not caused by a retrovirus. In certain embodiments, the
virally-induced condition is a cancer, an inflammatory condition,
an allergic condition, an autoimmune condition, or a skin
condition. In some embodiments, the virally-induced condition is a
lymphoma, chronic lymphocytic leukemia, nasopharyngeal carcinoma,
gastric cancer, Kaposi's sarcoma, rheumatoid arthritis, systemic
lupus erythematosus, or multiple sclerosis. In certain embodiments,
the virally-induced condition is not sepsis or viremia.
[0040] In some embodiments, the HDAC inhibitor can penetrate the
blood brain barrier. In certain embodiments, the antiviral agent is
a HIV drug, a Herpes drug, a CMV drug, or a hepatitis drug. In some
embodiments, the antiviral agent is acyclovir, ganciclovir or
valganciclovir. In certain embodiments, the antiviral agent is not
a heat shock protein inhibitor, an immunosuppressant, an
antibiotic, a glucocorticoid, a non-steroidal anti-inflammatory
drug, a Cox-2-specific inhibitor or a TNF-.alpha. binding protein.
In other embodiments, the antiviral agent is not a Hsp90 inhibitor,
tacrolimus, cyclosporin, rapamycin (sirolimus), methotrexate,
cyclophosphamide, azathioprine, mercaptopurine, mycophenolate,
FTY720, levofloxacin, amoxycillin, prednisone, cortisone acetate,
prednisolone, methylprednisolone, dexamethasone, betamethasone,
triamcinolone, beclometasone, fludrocortisone acetate,
deoxycorticosterone acetate, aldosterone, salicylates, arylalkanoic
acids, a 2-arylpropionic acid, a N-arylanthranilic acid, an oxicam,
a coxib, a sulphonanilide, valdecoxib, celecoxib, rofecoxib,
leflunomide, gold thioglucose, gold thiomalate, aurofin,
sulfasalazine, hydroxychloroquinine, minocycline, infliximab,
etanercept, adalimumab, abatacept, anakinra, interferon-.beta.,
interferon-.gamma., interleukin-2, an allergy vaccine, an
antihistamine, an antileukotriene, a beta-agonist, theophylline, or
an anticholinergic.
[0041] In some embodiments, the method further comprises
administering an additional agent. In certain embodiments, the
additional agent is an antiviral agent, a HDAC inhibitor, or a
chemotherapeutic therapy.
[0042] Further provided herein, in another aspect, is a composition
comprising a (i) HDAC inhibitor and (ii) an antiviral agent wherein
the HDAC inhibitor is a pyrimidine hydroxamic acid derivative. In
some embodiments, the HDAC inhibitor is JNJ-26481585
(N-hydroxy-2-(4-((((1-methyl-1H-indol-3-yl)methyl)amino)methyl)piperidin--
1-yl)pyrimidine-5-carboxamide), R306465/JNJ-16241199
(N-hydroxy-5-(4-(naphthalen-2-ylsulfonyl)piperazin-1-yl)pyrimidine-2-carb-
oxamide), or CHR-3996
(2-(6-{[(6-Fluoroquinolin-2-yl)methyl]amino}-3-azabicyclo[3.1.0]hex-3-yl)-
-N-hydroxypyrimidine-5-carboxamide). In some embodiments, the HDAC
inhibitor is not m-carboxycinnamic acid, bishydroxamic acid,
suberic bishydroxamic acid, Trichostatin A
(7-[4-(dimethylamino)phenyl]-N-hydroxy-4,6-dimethyl-7-oxohepta-2,4-dienam-
ide), SAHA (suberoyl anilide hydroxamic acid)/Vorinostat,
oxamflatin, ABHA, SB-55629, pyroxamide, propenamides, aroyl
pyrrolyl hydroxamides, Belinostat/PXD101, Papobinostat, LAQ824
(((E)-N-hydroxy-3-[4-[[2-hydroxyethyl-[2-(1H-indol-3-yl)ethyl]amino]methy-
l]phenyl]prop-2-enamide), LBH589, or TSA. In certain embodiments,
the HDAC inhibitor can penetrate the blood brain barrier.
[0043] In some embodiments, the antiviral agent is a HIV drug, a
Herpes drug, a CMV drug, or a hepatitis drug. In certain
embodiments, the antiviral agent is acyclovir, ganciclovir, or
valganciclovir. In some embodiments, the antiviral agent is not a
heat shock protein inhibitor, an immunosuppressant, an antibiotic,
a glucocorticoid, a non-steroidal anti-inflammatory drug, a
Cox-2-specific inhibitor or a TNF-.alpha. binding protein. In other
embodiments, the antiviral agent is not a Hsp90 inhibitor,
tacrolimus, cyclosporin, rapamycin (sirolimus), methotrexate,
cyclophosphamide, azathioprine, mercaptopurine, mycophenolate,
FTY720, levofloxacin, amoxycillin, prednisone, cortisone acetate,
prednisolone, methylprednisolone, dexamethasone, betamethasone,
triamcinolone, beclometasone, fludrocortisone acetate,
deoxycorticosterone acetate, aldosterone, salicylates, arylalkanoic
acids, a 2-arylpropionic acid, a N-arylanthranilic acid, an oxicam,
a coxib, a sulphonanilide, valdecoxib, celecoxib, rofecoxib,
leflunomide, gold thioglucose, gold thiomalate, aurofin,
sulfasalazine, hydroxychloroquinine, minocycline, infliximab,
etanercept, adalimumab, abatacept, anakinra, interferon-.beta.,
interferon-.gamma., interleukin-2, an allergy vaccine, an
antihistamine, an antileukotriene, a beta-agonist, theophylline, or
an anticholinergic.
[0044] In certain embodiments, the composition further comprises an
additional agent. In some embodiments, the additional agent is an
antiviral agent, a HDAC inhibitor, or a chemotherapeutic drug.
[0045] In some embodiments, the HDAC inhibitor and the antiviral
agent are in an oral formulation. In certain embodiments, the HDAC
inhibitor is present at 0.01-1 mg/kg per dose. In some embodiments,
the oral formulation comprises a unit dose of no greater than 80 mg
of the HDAC inhibitor and no greater than 1500 mg of the antiviral
agent.
[0046] Additionally provided herein, in a further aspect, is a
method for treating and/or preventing a virus-induced inflammatory
condition in a subject comprising administering a viral inducing
agent and an antiviral agent to the subject, thereby treating
and/or preventing the inflammatory condition. In some embodiments,
the virus is a member of the herpes virus family, human
immunodeficiency virus, parvovirus, or coxsackie virus. In certain
embodiments, the member of the herpes virus family is herpes
simplex virus, herpes genitalis virus, varicella zoster virus,
Epstein-Barr virus, human herpesvirus 6, or cytomegalovirus. In
some embodiments, the member of the herpes virus family is
Epstein-Barr virus, cytomegalovirus, or human herpesvirus 6.
[0047] In certain embodiments, the inflammatory condition is an
autoimmune condition. In some embodiments, the autoimmune condition
is rheumatoid arthritis, multiple sclerosis, Sjogren's syndrome,
systemic lupus erythematosus, autoimmune hepatitis, autoimmune
thyroiditis, hemophagocytic syndrome, diabetes, Crohn's condition,
ulcerative colitis, psoriasis, psoriatic arthritis, idiopathic
thrombocytonpenic pupura, polymyositis, dermatomyositis, myasthenia
gravis, autoimmune thryroiditis, Evan's syndrome, autoimmune
hemolytic anemia, aplastic anemia, autoimmune neutropenia,
scleroderma, Reiter's syndrome, ankylosing spondylitis, pemphnigus,
pemphigoid or autoimmune hepatitis. In certain embodiments, the
inflammatory condition is an allergic condition. In some
embodiments, the inflammatory condition is a skin condition. In
some embodiments, the inflammatory condition is associated with
coronary artery condition or peripheral artery condition. In
certain embodiments, the inflammatory condition is retinitis,
pancreatitis, cardiomyopathy, pericarditis, colitis,
glomerulonephritis, lung inflammation, esophagitis, gastritis,
duodenitis, ileitis, meningitis, encephalitis, encephalomyelitis,
transverse myelitis, cystitis, urethritis, mucositis,
lymphadenitis, dermatitis, hepatitis, osteomyelitis, or herpes
zoster. In some embodiments, the inflammatory condition is
atherosclerosis. In certain embodiments, the virus is
cytomegalovirus or herpes simplex virus.
[0048] In some embodiments, the viral inducing agent is one or more
of a chemotherapeutic drug, HDAC inhibitor, or DNA demethylating
agent. In certain embodiments, the HDAC inhibitor is butyrate or
MS-275.
[0049] In some embodiments, the viral inducing agent can penetrate
the blood brain barrier. In certain embodiments, the viral inducing
agent comprises arginine butyrate. In some embodiments, the
antiviral agent is ganciclovir or valganciclovir.
[0050] In certain embodiments, the method further comprises
administering an additional agent. In some embodiments, the
additional agent comprises a vaccine. In certain embodiments, the
vaccine comprises myelin basic protein and the condition is
multiple sclerosis. In some embodiments, the vaccine comprises an
antigen and the condition is diabetes. In certain embodiments, the
additional agent is aspirin, naproxen, ibuprofen, or a statin. In
some embodiments, the inflammatory condition is an autoimmune
condition and the additional agent is cyclosporine, azathiorprine,
methotrexate, cyclophosphamide, FK506, tacrolimus, monoclonal
antibody, anti-T cell monoclonal antibody, anti-B cell monoclonal
antibody, IL-2 receptor antibody, or a TNF inhibitor. In certain
embodiments, the monoclonal antibody is an anti-B cell antibody. In
some embodiments, the anti-B cell antibody is anti-CD20. In certain
embodiments, the anti-T cell antibody is an anti-CD3 antibody. In
some embodiments, the anti-CD3 antibody is OKT3. In certain
embodiments, the TNF inhibitor is infliximab (Remicade.TM.),
etanercept (Enbrel.TM.), Adalimumab (Humira.TM.), or an anti-IL-6
antibody.
[0051] In certain embodiments, the inflammatory condition is
atherosclerosis and the additional agent is a lipid lowering agent.
In some embodiments, the lipid lowering agent is rosuvastatin,
atorvastatin, simvastatin, or lovastatin. In certain embodiments,
the inflammatory condition is multiple sclerosis and the additional
agent is mitoxantrone, cladribine, or Campath antibody. In some
embodiments, the viral inducing agent is administered to the
subject before the antiviral agent.
INCORPORATION BY REFERENCE
[0052] All publications, patents, and patent applications mentioned
in this specification are herein incorporated by reference to the
same extent as if each individual publication, patent, or patent
application was specifically and individually indicated to be
incorporated by reference.
[0053] The following are also incorporated by reference: U.S. Pat.
No. 6,677,302; U.S. Pat. No. 7,399,787; US 2009/0270497; US
2010/0093824; US 2010/0152155; WO1998/04290; WO 2004/113336; WO
2008/097654; Moffat, D. et al, Discovery of
2-(6-{[(6-Fluoroquinolin-2-yl)methyl]amino}bicyclo[3.1.0]hex-3-yl)-N-hydr-
oxypyrimidine-5-carboxamide (CHR-3996), a Class I Selective Orally
Active HDAC inhibitor, Journal of Medicinal Chemistry (2010), 53,
8663-8678; Glaser, K B, HDAC inhibitors: Clinical update and
mechanisms-based potential, Biochem. Pharmacol. (2007); Ghosh, S.
K., et al. 2007 Blood Cells, Molecules, and Diseases 38:57-65.
BRIEF DESCRIPTION OF THE DRAWINGS
[0054] The novel features of the invention are set forth with
particularity in the appended claims. A better understanding of the
features and advantages of the present invention will be obtained
by reference to the following detailed description that sets forth
illustrative embodiments, in which the principles of the invention
are utilized, and the accompanying drawings of which:
[0055] FIGS. 1A-1B illustrate a CT scan of tumor reduction after
treatment with arginine butyrate (AB) and GCV. FIG. 1A and FIG. 1B
show pre- and post-treatment, respectively.
[0056] FIGS. 2A-2D illustrate results from toxicity assays with
anti-herpesvirus drugs ganciclovir (GCV) and penciclovir (PCV).
FIG. 2A and FIG. 2B show results from various concentrations of GCV
and PCV. FIG. 2C and FIG. 2D show results from GCV and PCV used in
combination with NaB.
[0057] FIGS. 3A-3C illustrate results from analysis of efficacy of
anti-virals using short chain fatty acids as inducing agents. FIG.
3A shows cell count results after using NaB. FIG. 3B shows results
from VA. FIG. 3C shows fold of TK expression induced.
[0058] FIGS. 4A-4B illustrate results from analysis of efficacy of
anti-virals using hydroxamic acids as inducing agents. FIG. 4A
shows cell count results after using scriptaid. FIG. 4B shows fold
of TK expression induced.
[0059] FIGS. 5A-5B illustrate results from analysis of efficacy of
anti-virals using SAHA (Vorinostat) as an inducing agent. FIG. 5A
shows cell count results after using SAHA. FIG. 5B shows fold of TK
expression induced.
[0060] FIGS. 6A-6B illustrate results from analysis of efficacy of
anti-virals using LHB589 (Panobinostat) as an inducing agent. FIG.
6A shows cell count results after using LHB589. FIG. 6B shows fold
of TK expression induced.
[0061] FIG. 7 illustrates results from analysis of efficacy of
anti-virals using PXD101, which induced a high level of TK
expression at the 5 .mu.M concentration.
[0062] FIG. 8 illustrates results from analysis of efficacy of
anti-virals using oxamflatin as an inducing agent. FIG. 8 shows
cell count results after using oxamflatin.
[0063] FIG. 9 illustrates results from analysis of efficacy of
anti-virals using a cyclic tetrapeptide as an inducing agent. FIG.
9 shows cell count results after using apicidin.
[0064] FIGS. 10A-10C illustrate results from analysis of efficacy
of anti-virals using a benzamide (MS-275) as an inducing agent.
FIG. 10A shows cell count results after using MS-275. FIG. 10B
shows fold of TK expression induced. FIG. 10C shows cell count
results after treating cells in combination for shorter time
periods.
[0065] FIGS. 11A-11B illustrate chemical structures of largazole
compounds used. Shown in both FIG. 11A and FIG. 11B.
[0066] FIGS. 12A-12F illustrate results from analysis of efficacy
of anti-virals using largazoles as an inducing agent. FIGS. 12A,
12B, 12C, 12D and 12E show cell count results after using various
largazole compounds. FIG. 12F shows fold of TK expression induced
from various largazole compounds.
[0067] FIG. 13 illustrates results from treatment of an
HIV-1-infected monocyte cell line with combination therapy. Viral
release (p24 release) was measured through optical density (OD)
measurement.
[0068] FIG. 14 illustrates results from treatment of an
HIV-1-infected monocyte cell line with combination therapy. Viral
release (p24 release) was measured through optical density (OD)
measurement and then converted into pg of protein.
[0069] FIG. 15 is a flow diagram of one embodiment of the provided
invention.
DETAILED DESCRIPTION OF THE INVENTION
[0070] Provided herein are methods and compositions for treating
and/or preventing viral conditions, virally-induced conditions, or
inflammatory conditions in a subject. The condition can be
associated with latent viral infections. The methods can comprise
the steps of administering a viral inducing agent and an antiviral
agent to the subject. The method can comprise steps of
administering a viral inducing agent, an antiviral agent, and one
or more additional agents to a subject. The methods include the
co-administration of an oral HDAC inhibitor and an antiviral agent,
either in the same or separate formulations.
[0071] The methods and compositions provided can be used to treat
and/or prevent infection by any of the viruses described herein.
The methods and compositions can be used to treat and/or prevent
any of the inflammatory conditions described herein. Any of the
viral inducing agents and/or antiviral agents described herein can
be used in the methods and compositions of the provided invention.
The viral inducing agent can be an HDAC inhibitor. The HDAC
inhibitor can be a pyrimidine hydroxamic acid derivative.
[0072] One or more additional agents described herein can be
administered to a subject. An additional agent can be selected for
administration based on the type of condition the subject has or is
suspected of having.
[0073] Another aspect of the present invention relates to
formulations, routes of administration and effective doses for
pharmaceutical compositions comprising an agent or combination of
agents, e.g., viral inducing agents, antiviral agents, or one or
more additional agents. A viral inducing agent, antiviral agent, or
one or more additional agents can be administered to a subject in
separate pharmaceutical compositions or can be co-formulated in a
single pharmaceutical composition.
[0074] Also provided are methods relating to dosing schedules for
administering a viral inducing agent, antiviral agent, or one or
more additional agents. One or more pharmaceutical compositions can
be administered to a subject by "pulsed administration" over a
period of time.
Overview
[0075] Like all other herpes viruses, EBV has two stages of
replication, the lytic and the latent. Soon after primary
infection, immunological surveillance by the host forces EBV to
enter the latent state of infection, where only few selected genes
are expressed. EBV maintains this latent state in all
EBV-associated tumors. Conventional anti-herpes virus drugs, such
as ganciclovir, acyclovir, etc., fail to act on these
latently-infected cells because the viral enzyme thymidine kinase
(TK) or protein kinase (PK), which is necessary for the conversion
of the prodrugs to their toxic metabolites, is not expressed in
latently-infected cells. Provided herein, in some embodiments, is a
combination treatment wherein lytic replication is induced and
antiviral drugs are administered concurrently.
[0076] Previous studies using patient-derived cells in vitro, and
also from phase I/II clinical studies on a series of patients with
EBV-associated lymphomas, have clearly shown the great promise of
this combination therapy approach. Strong epidemiological
association of Epstein-Barr Virus (EBV) with various human lymphoid
malignancies and in vitro studies demonstrating tumorigenic
activity of many EBV latent gene products suggest a causal
relationship between EBV and these diseases. However, as EBV
maintains a latent state of infection in these lymphomas, typical
anti-herpesviral drugs, such as the nucleoside analogs ganciclovir
(GCV) or acyclovir, are ineffective as these prodrugs require
expression of a lytic phase EBV protein, thymidine kinase (TK) or
protein kinase (EBV-PK), for their activity. Therefore, selective
induction of EBV lytic-phase gene expression in lymphoma cells that
harbor latent EBV, coupled with simultaneous exposure to antiviral
drugs, has been advanced as promising targeted therapy, because of
resulting targeting of cytotoxicity to the EBV-infected tumor
cells.
[0077] A variety of agents including short-chain fatty acids and
chemotherapeutic drugs, have been used to induce EBV lytic-phase
infection in cultured cells, but these in vitro studies have
generally not resulted in clinical application. For instance,
arginine butyrate and GCV has successfully been used to treat
EBV-positive lymphoid malignancies in a recent Phase I/II clinical
trial. In this study of 15 patients with relapsed or refractory
EBV-positive lymphoid tumors, 4 patients achieved complete tumor
remissions and 6 patients partial tumor remissions. However, the
rapid metabolism of butyrate requires continuous IV administration
of high doses. Butyrate has pan-HDAC inhibitory activity, and it
has been established that this activity is responsible for the
induction of the EBV-TK protein. HDAC inhibitors have been shown to
induce both EBV-TK and EBV-PK in EBV infected tumors.
[0078] In recent years, several potent HDAC inhibitors (HDACi) have
been tested in the clinic as anti-cancer agents. In certain
instances, HDAC inhibitors, including some new, highly-potent
compounds, induce EBV lytic phase gene expression and kill
EBV-infected cells in combination with antiviral drugs. In some
instances, HDAC inhibitors induce lytic phase gene expression in
viruses and kill virus-infected cells in combination with antiviral
drugs. HDAC inhibitors include, but are not limited to, short-chain
fatty acids (sodium butyrate and valproic acid), hydroxamic acids
(Oxamflatin, Scriptaid, CHR-3996, Suberoyl anilide hydroxamic acid
(SAHA), Panobinostat (LBH589) and Belinostat (PXD101)), the
benzamide MS275, cyclic tetrapeptide Apicidin, and newly-identified
HDAC inhibitor Largazole, which was originally isolated from a
marine cyanobacterium. In any of the embodiments herein, the HDAC
inhibitor is preferably suitable for oral administration.
Methods and Compositions
[0079] In one aspect, provided herein are methods for treating
and/or preventing a viral condition, a virally-induced condition,
or an inflammatory condition. In some embodiments, the condition is
associated with a latent viral infection. In certain embodiments,
the methods comprise administering a viral inducing agent (e.g., an
HDAC inhibitor) and an antiviral agent. In some embodiments, the
methods comprise administering an HDAC inhibitor and an antiviral
agent. In certain embodiments, the HDAC inhibitor and the antiviral
agent are co-formulated. In some embodiments, the methods comprise
further administering an additional viral inducing agent. In other
embodiments, the methods comprise further administering an
additional antiviral agent. In some embodiments, the methods
comprise administering additional individual doses of the viral
inducing agent and/or the antiviral agent.
[0080] Further provided herein are methods for treating and/or
preventing a viral condition, a virally-induced condition, or an
inflammatory condition comprising administering an HDAC inhibitor.
In some embodiments, the HDAC inhibitor is a pyrimidine hydroxamic
acid derivative. In certain embodiments, the HDAC inhibitor is a
pyrimidine hydroxamic acid derivative not containing a piperidine,
or a pyrimidine hydroxamic acid derivative comprising an
azabicyclohexane. In certain embodiments, the methods further
comprise administering an antiviral agent. In some embodiments, the
HDAC inhibitor and the antiviral agent are co-formulated.
[0081] Also provided herein are methods for treating and/or
preventing a viral or virally-induced condition, or an inflammatory
condition comprising administering an HDAC inhibitor and an
antiviral agent. In some embodiments, the viral or virally-induced
condition is caused by a DNA virus. In certain embodiments, the
HDAC inhibitor is administered at a dose of less than 2 mg/kg per
dose. In some embodiments, the HDAC inhibitor is administered at a
dose of less than 20 mg/kg, less than 19 mg/kg, less than 18 mg/kg,
less than 17 mg/kg, less than 16 mg/kg, less than 15 mg/kg, less
than 14 mg/kg, less than 13 mg/kg, less than 12 mg/kg, less than 11
mg/kg, less than 10 mg/kg, less than 9 mg/kg, less than 8 mg/kg,
less than 7 mg/kg, less than 6 mg/kg, less than 5 mg/kg, less than
4 mg/kg, less than 3 mg/kg, less than 2 mg/kg, less than 1 mg/kg,
less than 0.5 mg/kg, less than 0.2 mg/kg, or less than 0.1
mg/kg.
[0082] Further provided herein are methods for treating and/or
preventing a viral condition, a virally-induced condition, or an
inflammatory condition comprising administering an HDAC inhibitor
and an antiviral agent wherein the HDAC inhibitor has a molecular
weight of greater than 275 g/mol. In some embodiments, the HDAC
inhibitor has a molecular weight of greater than 200 g/mol, greater
than 225 g/mol, greater than 250 g/mol, greater than 275 g/mol,
greater than 300 g/mol, greater than 325 g/mol, greater than 350
g/mol, greater than 375 g/mol, greater than 400 g/mol, greater than
425 g/mol, greater than 450 g/mol, greater than 475 g/mol, greater
than 500 g/mol, greater than 525 g/mol, greater than 550 g/mol,
greater than 575 g/mol, greater than 600 g/mol, greater than 625
g/mol, greater than 650 g/mol, greater than 675 g/mol, greater than
700 g/mol, greater than 725 g/mol, greater than 750 g/mol, greater
than 775 g/mol, greater than 800 g/mol, greater than 850 g/mol,
greater than 900 g/mol, greater than 950 g/mol, or greater than
1000 g/mol. In certain embodiments, the HDAC inhibitor has a
molecular weight of less than 200 g/mol, less than 225 g/mol, less
than 250 g/mol, less than 275 g/mol, less than 300 g/mol, less than
325 g/mol, less than 350 g/mol, less than 375 g/mol, less than 400
g/mol, less than 425 g/mol, less than 450 g/mol, less than 475
g/mol, less than 500 g/mol, less than 525 g/mol, less than 550
g/mol, less than 575 g/mol, less than 600 g/mol, less than 625
g/mol, less than 650 g/mol, less than 675 g/mol, less than 700
g/mol, less than 725 g/mol, less than 750 g/mol, less than 775
g/mol, less than 800 g/mol, less than 850 g/mol, less than 900
g/mol, less than 950 g/mol, or less than 1000 g/mol. In some
embodiments, the HDAC inhibitor has a molecular weight of less than
500 g/mol and more than 250 g/mol. In other embodiments, the HDAC
inhibitor has a molecular weight of less than 400 g/mol and more
than 300 g/mol. In some instances, the HDAC inhibitor is not
butyric acid or arginine butyrate.
[0083] Also provided herein are methods for treating and/or
preventing Epstein-Barr virus (EBV) associated lymphoma. In some
embodiments, the methods comprise administering an HDAC inhibitor
and acyclovir and/or ganciclovir and/or valganciclovir.
[0084] Further provided here are methods for treating and/or
preventing a virus-induced inflammatory condition. In some
embodiments, the methods comprise administering a viral inducing
agent and an antiviral agent. In certain embodiments the virus is a
member of the herpes family (e.g., herpes simplex virus, herpes
genitalis virus, varicella zoster virus, Epstein-Barr virus, human
herpes virus 6, or cytomegalovirus)., human immunodeficiency virus,
parvovirus, or coxsackie virus.
[0085] In another aspect, provided herein are compositions
comprising an HDAC inhibitor and an antiviral agent. In certain
embodiments, the HDAC inhibitor is a pyrimidine hydroxamic acid
derivative. In some embodiments, the HDAC inhibitor is
JNJ-26481585, JNJ-16241199, or CHR-3996. In some embodiments, the
antiviral agent is acyclovir, ganciclovir, or valganciclovir. In
certain embodiments, the composition comprises an additional agent.
In some embodiments, the additional agent is a antiviral agent, an
HDAC inhibitor, or a chemotherapeutic agent. In certain
embodiments, the compositions are formulated as a capsule, gel,
tablet, solution, or suspension. In some embodiments, the
compositions are formulated for oral administration. In other
embodiments, the compositions are formulated for parenteral
administration. In some embodiments, the compositions are
formulated for intravenous, intraperitoneal, oral, subcutaneous,
intrathecal, or intratumoral administration. In certain
embodiments, the compositions are formulated for administration at
the site of a viral infection. In some embodiments, the
compositions are formulated for modified release of the HDAC
inhibitor and the antiviral agent. In specific embodiments, the
HDAC inhibitor is dissolved before the antiviral agent is
dissolved. In other specific embodiments, the HDAC inhibitor is
dissolved after the antiviral agent is dissolved. In some
embodiments, the compositions are formulated for once daily
administration. In other embodiments, the compositions are
formulated for twice daily, thrice daily, four times daily, once
every other day, once weekly, once bi-weekly, or monthly.
DEFINITIONS
[0086] The terms "viral," "virus-associated," and "virally-induced"
with reference to disorders are used interchangeably throughout the
instant specification.
[0087] The term "obtaining" as in "obtaining the composition" is
intended to include purchasing, synthesizing, or otherwise
acquiring the composition (or agent(s) of the composition).
[0088] The terms "comprises", "comprising", are intended to have
the broad meaning ascribed to them and can mean "includes",
"including" and the like.
[0089] The term "subject", "patient" or "individual" are used
interchangeably herein and refer to mammals and non-mammals, e.g.,
suffering from a disorder described herein. Examples of mammals
include, but are not limited to, any member of the Mammalian class:
humans, non-human primates such as chimpanzees, and other apes and
monkey species; farm animals such as cattle, horses, sheep, goats,
swine; domestic animals such as rabbits, dogs, and cats; laboratory
animals including rodents, such as rats, mice and guinea pigs, and
the like. Examples of non-mammals include, but are not limited to,
birds, fish and the like. In one embodiment of the methods and
compositions provided herein, the mammal is a human.
[0090] The terms "treat," "treating" or "treatment," and other
grammatical equivalents as used herein, include alleviating,
inhibiting or reducing symptoms, reducing or inhibiting severity
of, reducing incidence of, prophylactic treatment of, reducing or
inhibiting recurrence of, delaying onset of, delaying recurrence
of, abating or ameliorating a disease or condition symptoms,
ameliorating the underlying metabolic causes of symptoms,
inhibiting the disease or condition, e.g., arresting the
development of the disease or condition, relieving the disease or
condition, causing regression of the disease or condition,
relieving a condition caused by the disease or condition, or
stopping the symptoms of the disease or condition. The terms
further include achieving a therapeutic benefit. By therapeutic
benefit is meant eradication or amelioration of the underlying
disorder being treated, and/or the eradication or amelioration of
one or more of the physiological symptoms associated with the
underlying disorder such that an improvement is observed in the
patient.
[0091] The terms "prevent," "preventing" or "prevention," and other
grammatical equivalents as used herein, include preventing
additional symptoms, preventing the underlying metabolic causes of
symptoms, inhibiting the disease or condition, e.g., arresting the
development of the disease or condition and are intended to include
prophylaxis. The terms further include achieving a prophylactic
benefit. For prophylactic benefit, the compositions are optionally
administered to a patient at risk of developing a particular
disease, to a patient reporting one or more of the physiological
symptoms of a disease, or to a patient at risk of reoccurrence of
the disease.
[0092] The terms "effective amount" or "therapeutically effective
amount" as used herein, refer to a sufficient amount of at least
one agent being administered which achieve a desired result, e.g.,
to relieve to some extent one or more symptoms of a disease or
condition being treated. In certain instances, the result is a
reduction and/or alleviation of the signs, symptoms, or causes of a
disease, or any other desired alteration of a biological system. In
certain instances, an "effective amount" for therapeutic uses is
the amount of the composition comprising an agent as set forth
herein required to provide a clinically significant decrease in a
disease. An appropriate "effective" amount in any individual case
is determined using any suitable technique, such as a dose
escalation study.
[0093] The terms "administer," "administering", "administration,"
and the like, as used herein, refer to the methods that are used to
enable delivery of agents or compositions to the desired site of
biological action. These methods include, but are not limited to
oral routes, intraduodenal routes, parenteral injection (including
intravenous, subcutaneous, intraperitoneal, intramuscular,
intravascular or infusion), topical and rectal administration.
Administration techniques that in some instances are employed with
the agents and methods described herein include, e.g., as discussed
in Goodman and Gilman, The Pharmacological Basis of Therapeutics
(current edition), Pergamon; and Remington's, Pharmaceutical
Sciences (current edition), Mack Publishing Co., Easton, Pa. In
certain embodiments, the agents and compositions described herein
are administered orally. In some embodiments, the compositions
described herein are administered parenterally.
[0094] The term "pharmaceutically acceptable" as used herein,
refers to a material that does not abrogate the biological activity
or properties of the agents described herein, and is relatively
nontoxic (i.e., the toxicity of the material significantly
outweighs the benefit of the material). In some instances, a
pharmaceutically acceptable material is administered to an
individual without causing significant undesirable biological
effects or significantly interacting in a deleterious manner with
any of the components of the composition in which it is
contained.
[0095] The term "pharmaceutically acceptable excipient," as used
herein, refers to carriers and vehicles that are compatible with
the active ingredient (for example, a compound of the invention) of
a pharmaceutical composition of the invention (and preferably
capable of stabilizing it) and not deleterious to the subject to be
treated. For example, solubilizing agents that form specific, more
soluble complexes with the compounds of the invention can be
utilized as pharmaceutical excipients for delivery of the
compounds. Suitable carriers and vehicles are known to those of
extraordinary skill in the art. The term "excipient" as used herein
will encompass all such carriers, adjuvants, diluents, solvents, or
other inactive additives. Suitable pharmaceutically acceptable
excipients include, but are not limited to, water, salt solutions,
alcohol, vegetable oils, polyethylene glycols, gelatin, lactose,
amylose, magnesium stearate, talc, silicic acid, viscous paraffin,
perfume oil, fatty acid monoglycerides and diglycerides,
petroethral fatty acid esters, hydroxymethyl-cellulose,
polyvinylpyrrolidone, etc. The pharmaceutical compositions of the
invention can also be sterilized and, if desired, mixed with
auxiliary agents, e.g., lubricants, preservatives, stabilizers,
wetting agents, emulsifiers, salts for influencing osmotic
pressure, buffers, colorings, flavorings and/or aromatic substances
and the like, which do not deleteriously react with the active
compounds of the invention.
[0096] The invention can be understood more fully by reference to
the following detailed description and illustrative examples, which
are intended to exemplify non-limiting embodiments of the
invention.
Viral Inducing Agents
[0097] The methods of the provided invention comprise use of one or
more pharmaceutical compositions provided herein comprising an
inducing agent to induce expression of a gene product in a
virus-infected cell. The gene product expressed can be a viral
enzyme or a cellular enzyme or activity that is largely expressed
in virus-infected cells. Expression products that can be targeted
include enzymes involved with DNA replication, for example, for
repair or replication of the genome, assembly of complete virus
particles, generation of viral membrane or walls, RNA transcription
or protein translation or combinations of these activities.
Interference with these processes can be performed by inducing and
then acting on an enzyme and, preferably, a critical enzyme in the
process. Inducing agents that can be used in the methods and
compositions of the provided invention are described, for example,
in U.S. Pat. Nos. 6,197,743 and 6,677,302, which are herein
incorporated by reference in their entireties.
[0098] Inducing agents according to the methods or compositions
provided herein include, without limitation, short-chain fatty acid
(SCFA) derivatives, histone deacetylase (HDAC) inhibitors, phorbol
esters, anticancer agents, and cytokines. In some embodiments, the
viral inducing agent is a chemotherapeutic drug, an HDAC inhibitor,
or a DNA demethylating agent.
[0099] In some embodiments, the inducing agent is a SCFA
derivative. Examples of SCFA inducing agents include propionic
acid, butyric acid, succinic acid, fumaric acid monoethyl ester,
dimethyl butyric acid, trifluorobutanol, chloropropionic acid,
isopropionic acid, 2-oxypentanoic acid, 2,2- or 3,3-dimethyl
butyric acid, 2,2- or 3,3-diethyl butyric acid, butyric acid ethyl
ester, 2-methyl butanoic acid, fumaric acid, and amides and salts
thereof. Other examples include methoxy acetic acid, methoxy
propionic acid, N-acetylglycine, mercaptoacetic acid, 1- or
2-methyl cyclopropane carboxylic acid, squaric acid, 2- or
3-phenoxy propionic acid, methoxy butyric acid, phenoxy acetic
acid, 2- or 3-phenoxy butyric acid, phenyl acetic acid, phenyl
propionic acid, 3-phenyl butyric acid, ethyl-phenyl acetic acid,
4-chloro-2-phenoxy-2-propionic acid, n-dimethyl butyric acid
glycine amide, o-benzoyl lactic acid, o-dimethyl butyric acid
lactate, cinnamic acid, dihydrocinnamic acid (C.sub.6 H.sub.5
CHCH.sub.3 COOH), alpha-methyl-dihydrocinnamic acid, thiophenoxy
acetic acid, and amines, amides and salts of these chemicals.
Useful amines and amides can include isobutylhydroxylamine, fumaric
acid monoamide, fumaramide, succinamide, or isobutyramide.
[0100] In other embodiments, inducing agents include retinoic acid,
retinol, cytosine arabinoside, phorbols such as the phorbol diester
12-0-tetradecanoylphorbol 13-acetate (TPA), teleocidine B, indole
alkaloids, cytotoxin, plant lectins from Streptomyces,
glucocorticoids such as estrogen and progesterone,
phytohemagglutinin (PHA), bryostatin, growth factors (e.g. PDGF,
VEGF, EGF, FGF, NGF, TGF, BCGF), anti-sense nucleic acids (e.g.
DNA, RNA or PNA), aptamers (nucleic acid oligonucleotides with
secondary or tertiary structures which bind with high affinity and
selectivity to a target molecule), erythropoietin (EPO), the
interleukins (IL-1, IL-2, IL-3, etc.), cAMP and cAMP analogs such
as dibutyryl cAMP, activin, inhibin, steel factor, interferon, the
bone morphogenic proteins (BMBs), hydroxyurea and dimethyl
sulfoxide (DMSO). Other inducing agents include interferons (e.g.
.alpha.-, .beta.-, .gamma.-interferon), cytokines such as tumor
necrosis factor (TNF), cell receptors, and growth factor
antagonists, which may be purified or recombinantly produced.
[0101] In some embodiments, the inducing agent is a anticancer
agent. In certain embodiments, the anticancer agent is a
chemotherapeutic anticancer agent. Examples of chemotherapeutic
anticancer agents include Nitrogen Mustards like bendamustine,
chlorambucil, chlormethine, cyclophosphamide, ifosfamide,
melphalan, prednimustine, trofosfamide; Alkyl Sulfonates like
busulfan, mannosulfan, treosulfan; Ethylene Imines like carboquone,
thiotepa, triaziquone; Nitrosoureas like carmustine, fotemustine,
lomustine, nimustine, ranimustine, semustine, streptozocin;
Epoxides like etoglucid; Other Alkylating Agents like dacarbazine,
mitobronitol, pipobroman, temozolomide; Folic Acid Analogues like
methotrexate, permetrexed, pralatrexate, raltitrexed; Purine
Analogs like cladribine, clofarabine, fludarabine, mercaptopurine,
nelarabine, tioguanine; Pyrimidine Analogs like azacitidine,
capecitabine, carmofur, cytarabine, decitabine, fluorouracil,
gemcitabine, tegafur; Vinca Alkaloids like vinblastine,
vincristine, vindesine, vinflunine, vinorelbine; Podophyllotoxin
Derivatives like etoposide, teniposide; Colchicine derivatives like
demecolcine; Taxanes like docetaxel, paclitaxel, paclitaxel
poliglumex; Other Plant Alkaloids and Natural Products like
trabectedin; Actinomycines like dactinomycin; Antracyclines like
aclarubicin, daunorubicin, doxorubicin, epirubicin, idarubicin,
mitoxantrone, pirarubicin, valrubicin, zorubincin; Other Cytotoxic
Antibiotics like bleomycin, ixabepilone, mitomycin, plicamycin;
Platinum Compounds like carboplatin, cisplatin, oxaliplatin,
satraplatin; Methylhydrazines like procarbazine; Sensitizers like
aminolevulinic acid, efaproxiral, methyl aminolevulinate, porfimer
sodium, temoporfin; Protein Kinase Inhibitors like dasatinib,
erlotinib, everolimus, zotarolimus, gefitinib, imatinib, lapatinib,
nilotinib, pazonanib, sorafenib, sunitinib, temsirolimus; Other
Antineoplastic Agents like alitretinoin, altretamine, amzacrine,
anagrelide, arsenic trioxide, asparaginase, bexarotene, bortezomib,
celecoxib, denileukin diftitox, estramustine, hydroxycarbamide,
irinotecan, lonidamine, masoprocol, miltefosein, mitoguazone,
mitotane, oblimersen, pegaspargase, pentostatin, romidepsin,
sitimagene ceradenovec, tiazofurine, topotecan, tretinoin,
vorinostat; Estrogens like diethylstilbenol, ethinylestradiol,
fosfestrol, polyestradiol phosphate; Progestogens like gestonorone,
medroxyprogesterone, megestrol; Gonadotropin Releasing Hormone
Analogs like buserelin, goserelin, leuprorelin, triptorelin;
Anti-Estrogens like fulvestrant, tamoxifen, toremifene;
Anti-Androgens like bicalutamide, flutamide, nilutamide; Enzyme
Inhibitors like aminoglutethimide, anastrozole, exemestane,
formestane, letrozole, vorozole; Other Hormone Antagonists like
abarelix, degarelix; Immunostimulants like histamine
dihydrochloride, mifamurtide, pidotimod, plerixafor, roquinimex,
thymopentin; Immunosuppressants like everolimus, zotarolimus,
gusperimus, leflunomide, mycophenolic acid, sirolimus; Calcineurin
Inhibitors like ciclosporin, tacrolimus; Other Immunosuppressants
like azathioprine, lenalidomide, methotrexate, thalidomide; and
Radiopharmaceuticals like iobenguane.
[0102] In certain embodiments, the viral inducing agent is a
demethylating agent. For example, demethylating agents include
decitabine and azacytidine. In other embodiments, the inducing
agent is a chemotherapy drug, such as cyclophosphamide, cisplatin,
melphalan, doxorubicin, daunorubicin, vincristine, methotrexate,
cytarabine, ifosfamide, etoposide, or rituximab.
[0103] In further embodiments, inducing agents include histone
deacetylase (HDAC) inhibitors (including those of the hydroxamic
acid class and the benzamide class), DNA methyltransferase
inhibitors, and proteasome inhibitors. HDAC inhibitors, a class of
compounds that interfere with the function of histone deacetylase,
include, without limitation, short-chain fatty acids (butyrate,
phenylbutyrate, valproate, AN-9, etc., as described above),
hydroxamic acids (for example m-carboxycinnamic acid, bishydroxamic
acid, suberic bishydroxamic acid, Trichostatin A
(7-[4-(dimethylamino)phenyl]-N-hydroxy-4,6-dimethyl-7-oxohepta-2,4-dienam-
ide), SAHA (suberoyl anilide hydroxamic acid)/Vorinostat,
oxamflatin, ABHA, SB-55629, pyroxamide, propenamides, aroyl
pyrrolyl hydroxamides, Belinostat/PXD101, Papobinostat, LAQ824
(((E)-N-hydroxy-3-[4-[[2-hydroxyethyl-[2-(1H-indol-3-yl)ethyl]amino]methy-
l]phenyl]prop-2-enamide), LBH589, CHR-3996
(2-(6-{[(6-Fluoroquinolin-2-yl)methyl]amino}-3-azabicyclo[3.1.0]hex-3-yl)-
-N-hydroxypyrimidine-5-carboxamide, TSA), Pivanex, spiruchostatins,
cyclic tetrapeptides (for example, trapoxin A
(cyclo((S)-phenylalanyl-(S)-phenylalanyl-(R)-pipecolinyl-(2S,9S)-2-amino--
8-oxo-9,10-epoxydecanoyl),), trapoxin B
(cyclo((S)-phenylalanyl-phenylalanyl-(R)-prolyl-2-amino-8-oxo-9,10-epoxyd-
ecanoyl)), HC-toxin, chlamydocin, diheteropeptin, WF-3161, Cyl-1,
Cyl-2, azumamide A), cyclic peptides (for example, FK-228,
FR901228), depsipeptides (for example, romidepsin, FK228
((E)-(1S,4S,10S,21R)-7[(Z)-ethylideno]-4,21-diisopropyl-2-oxa-12,13-dithi-
a-5,8,20,23-tetraazabicyclo[8,7,6]-tricos-16-ene-3,6,9,22-pentanone),
FK228 analogs and derivatives, largazole, largazole analogs and
derivatives), peptide antibiotics (apicidin), benzamides (MS275
(3-pyridinylmethyl[[4-[[(2-aminophenyl)amino]carbonyl]phenyl]methyl]carba-
mate,
N-(2-Aminophenyl)-4-[N-(pyridine-3ylmethoxycarbonyl)aminomethyl]benz-
amide), CI994 (4-(Acetylamino)-N-(2-aminophenyl)benzamide),
MGCD0103), electrophilic ketones (TPX, AOR, Depudecin), FR901375,
nicotinamide, NAD derivatives, Sirtinol, splitomycin,
dihydrocoumarin, naphthopyranone, 2-hydroxynaphthaldehydes,
PCYC-0402, PCYC-0403, PCI-24781
(3-(dimethylaminomethyl)-N-[2-[4-(hydroxycarbamoyl)phenoxy]ethyl]-1-benzo-
furan-2-carboxamide), depudecin, tubacin, organosulfur compounds,
and dimethyl sulfoxide (DMSO). Other compounds which may also be
administered as inducing agents, which include CHAPs, Scriptaid,
Tubacin, JNJ16241199, A-161906, 6-(3-Chlorophenylureido)caproic
hydroxamic acid, SB939, ITF2357
({6-[(diethylamino)methyl]-2-naphthyl}methyl
{4-[(hydroxyamino)carbonyl]phenyl}carbamate), 4SC-201, AR-42,
OPB-801, RG2833, CUDC-101, JNJ-26481585, MK0683 (suberoylanilide
hydroxamic acid), M344
(4-(Diethylamino)-N-[7-(hydroxyamino)-7-oxoheptyl]benzamide),
BML-210 (N-(2-aminophenyl)-N'-phenyl-octanediamide), dacinostat
(NVP-LAQ824), PDX-101, BAY86-5274, SB939, droxinostat, and
pivaloyloxymethyl butyrate.
[0104] In some embodiments, the viral inducing agent is an HDAC
inhibitor. In some embodiments, the HDAC inhibitor is a hydroxamic
acid, for example, Vorinostat/suberoyl anilide hydroxamic acid
(SAHA), JNJ-26481585
(N-hydroxy-2-(4-((((1-methyl-1H-indol-3-yl)methyl)amino)methyl)piperidin--
1-yl)pyrimidine-5-carboxamide), R306465/JNJ-16241199
(N-hydroxy-5-(4-(naphthalen-2-ylsulfonyl)piperazin-1-yl)pyrimidine-2-carb-
oxamide), CHR-3996
(2-(6-{[(6-Fluoroquinolin-2-yl)methyl]amino}-3-azabicyclo[3.1.0]hex-3-yl)-
-N-hydroxypyrimidine-5-carboxamide), Belinostat/PXD101,
Panobinostat/LBH-589, trichostatin A/TSA
(7-[4-(dimethylamino)phenyl]-N-hydroxy-4,6-dimethyl-7-oxohepta-2,4-dienam-
ide), ITF2357, CBHA, and Givinostat/ITF2357. In certain
embodiments, the HDAC inhibitor is a pyrimidine hydroxamic acid,
for example, JNJ-26481585, JNJ-16241199, or CHR-3996. In other
embodiments, the HDAC inhibitor is a benzamide, for example,
CI-994, Entinostat/MS-275, SNDX-275, and mocetinostat/MGCD0103
(N-(2-aminophenyl)-4-((4-pyridin-3-ylpyrimidin-2-ylamino)methyl)benzamide-
).
[0105] In some embodiments, the HDAC inhibitor is a hydroxamic acid
derivative. In certain embodiments, the HDAC inhibitor is a
pyrimidine hydroxamic acid. In some embodiments, the HDAC inhibitor
is a non-piperidine-containing pyrimidine hydroxamic acid
derivative. In certain embodiments, the HDAC inhibitor comprises an
azabicyclo-hexane. In other embodiments, the HDAC inhibitor
comprises fluorine. In certain embodiments, the HDAC inhibitor
comprises a fluoroquinoline group.
[0106] In one embodiment the HDAC inhibitor is JNJ-26481585
(N-hydroxy-2-(4-((((1-methyl-1H-indol-3-yl)methyl)amino)methyl)piperidin--
1-yl)pyrimidine-5-carboxamide). In a further embodiment, the HDAC
inhibitor is R306465/JNJ-16241199
(N-hydroxy-5-(4-(naphthalen-2-ylsulfonyl)piperazin-1-yl)pyrimidine-2-carb-
oxamide). In a specific embodiment, the HDAC inhibitor is CHR-3996
[2-(6-{[(6-Fluoroquinolin-2-yl)methyl]amino}-3-azabicyclo[3.1.0]hex-3-yl)-
-N-hydroxypyrimidine-5-carboxamide, or N-hydroxy
2-(6-{[(6-Fluoroquinolin-2-yl)methyl]amino}-3-azabicyclo[3.1.0]hex-3-yl)--
pyrimidine-5-carboxamide].
[0107] In another embodiment, the molecular weight of the HDAC
inhibitor is greater than 275 g/mol. In some embodiments, the HDAC
inhibitor has a molecular weight of greater than 200 g/mol, greater
than 225 g/mol, greater than 250 g/mol, greater than 275 g/mol,
greater than 300 g/mol, greater than 325 g/mol, greater than 350
g/mol, greater than 375 g/mol, greater than 400 g/mol, greater than
425 g/mol, greater than 450 g/mol, greater than 475 g/mol, greater
than 500 g/mol, greater than 525 g/mol, greater than 550 g/mol,
greater than 575 g/mol, greater than 600 g/mol, greater than 625
g/mol, greater than 650 g/mol, greater than 675 g/mol, greater than
700 g/mol, greater than 725 g/mol, greater than 750 g/mol, greater
than 775 g/mol, greater than 800 g/mol, greater than 850 g/mol,
greater than 900 g/mol, greater than 950 g/mol, or greater than
1000 g/mol. In certain embodiments, the HDAC inhibitor has a
molecular weight of less than 200 g/mol, less than 225 g/mol, less
than 250 g/mol, less than 275 g/mol, less than 300 g/mol, less than
325 g/mol, less than 350 g/mol, less than 375 g/mol, less than 400
g/mol, less than 425 g/mol, less than 450 g/mol, less than 475
g/mol, less than 500 g/mol, less than 525 g/mol, less than 550
g/mol, less than 575 g/mol, less than 600 g/mol, less than 625
g/mol, less than 650 g/mol, less than 675 g/mol, less than 700
g/mol, less than 725 g/mol, less than 750 g/mol, less than 775
g/mol, less than 800 g/mol, less than 850 g/mol, less than 900
g/mol, less than 950 g/mol, or less than 1000 g/mol. In some
embodiments, the HDAC inhibitor has a molecular weight of less than
500 g/mol and more than 250 g/mol. In other embodiments, the HDAC
inhibitor has a molecular weight of less than 400 g/mol and more
than 300 g/mol.
[0108] In a particular embodiment, the HDAC inhibitor is not
m-carboxycinnamic acid, bishydroxamic acid, suberic bishydroxamic
acid, Trichostatin A
(7-[4-(dimethylamino)phenyl]-N-hydroxy-4,6-dimethyl-7-oxohepta-2,4-dienam-
ide), SAHA (suberoyl anilide hydroxamic acid)/Vorinostat,
oxamflatin, ABHA, SB-55629, pyroxamide, propenamides, aroyl
pyrrolyl hydroxamides, Belinostat/PXD101, Papobinostat, LAQ824
(((E)-N-hydroxy-3-[4-[[2-hydroxyethyl-[2-(1H-indol-3-yl)ethyl]amino]methy-
l]phenyl]prop-2-enamide), or LBH589.
[0109] In some embodiments, a viral inducing agent, for example an
HDAC inhibitor, penetrates the blood brain barrier. In certain
embodiments, a viral inducing agent that penetrates the blood brain
barrier comprises arginine butyrate, SAHA, or CHR-3996.
[0110] In certain embodiments, the HDAC inhibitor is administered
at a dose of less than 400 mg/day. In some embodiments, the HDAC
inhibitor is administered at a dose of about 1 mg/day, about 2
mg/day, about 5 mg/day, about 10 mg/day, about 15 mg/day, about 20
mg/day, about 25 mg/day, about 30 mg/day, about 35 mg/day, about 40
mg/day, about 45 mg/day, about 50 mg/day, about 60 mg/day, about 70
mg/day, about 80 mg/day, about 90 mg/day, about 100 mg/day, about
120 mg/day, about 125 mg/day, about 140 mg/day, about 150 mg/day,
about 160 mg/day, about 175 mg/day, about 180 mg/day, about 190
mg/day, about 200 mg/day, about 225 mg/day, about 250 mg/day, about
275 mg/day, about 300 mg/day, about 325 mg/day, about 350 mg/day,
about 375 mg/day, about 400 mg/day, about 425 mg/day, about 450
mg/day, about 475 mg/day, or about 500 mg/day. In certain
embodiments, the HDAC inhibitor is administered at a dose of less
than 1 mg/day, less than 2 mg/day, less than 5 mg/day, less than 10
mg/day, less than 15 mg/day, less than 20 mg/day, less than 25
mg/day, less than 30 mg/day, less than 35 mg/day, less than 40
mg/day, less than 45 mg/day, less than 50 mg/day, less than 60
mg/day, less than 70 mg/day, less than 80 mg/day, less than 90
mg/day, less than 100 mg/day, less than 120 mg/day, less than 125
mg/day, less than 140 mg/day, less than 150 mg/day, less than 160
mg/day, less than 175 mg/day, less than 180 mg/day, less than 190
mg/day, less than 200 mg/day, less than 225 mg/day, less than 250
mg/day, less than 275 mg/day, less than 300 mg/day, less than 325
mg/day, less than 350 mg/day, less than 375 mg/day, less than 400
mg/day, less than 425 mg/day, less than 450 mg/day, less than 475
mg/day, or less than 500 mg/day. In some embodiments, the HDAC
inhibitor is administered at a dose of more than 1 mg/day, more
than 2 mg/day, more than 5 mg/day, more than 10 mg/day, more than
15 mg/day, more than 20 mg/day, more than 25 mg/day, more than 30
mg/day, more than 35 mg/day, more than 40 mg/day, more than 45
mg/day, more than 50 mg/day, more than 60 mg/day, more than 70
mg/day, more than 80 mg/day, more than 90 mg/day, more than 100
mg/day, more than 120 mg/day, more than 125 mg/day, more than 140
mg/day, more than 150 mg/day, more than 160 mg/day, more than 175
mg/day, more than 180 mg/day, more than 190 mg/day, more than 200
mg/day, more than 225 mg/day, more than 250 mg/day, more than 275
mg/day, more than 300 mg/day, more than 325 mg/day, more than 350
mg/day, more than 375 mg/day, more than 400 mg/day, more than 425
mg/day, more than 450 mg/day, more than 475 mg/day, or more than
500 mg/day. In certain embodiments, the HDAC inhibitor is
administered at a dose of more than 1 mg/day and less than 500
mg/day. In some embodiments, the HDAC inhibitor is administered at
a dose of more than 20 mg/day and less than 80 mg/day. In certain
embodiments, the HDAC inhibitor is administered once a day (q.d.),
twice a day (b.id.), or thrice a day (t.i.d.). In some embodiments,
the HDAC inhibitor is administered daily, once a week, twice a
week, three times a week, four times a week, or five times a
week.
[0111] In some embodiments, the HDAC inhibitor is CHR-3996. In
certain embodiments, CHR-3996 is administered at a dose of 40
mg/day. In some embodiments, CHR-3996 is administered at a dose of
about 1 mg/day, about 2 mg/day, about 5 mg/day, about 10 mg/day,
about 15 mg/day, about 20 mg/day, about 25 mg/day, about 30 mg/day,
about 35 mg/day, about 40 mg/day, about 45 mg/day, about 50 mg/day,
about 60 mg/day, about 70 mg/day, about 80 mg/day, about 90 mg/day,
or about 100 mg/day. In certain embodiments, CHR-3996 is
administered at a dose of less than 1 mg/day, less than 2 mg/day,
less than 5 mg/day, less than 10 mg/day, less than 15 mg/day, less
than 20 mg/day, less than 25 mg/day, less than 30 mg/day, less than
35 mg/day, less than 40 mg/day, less than 45 mg/day, less than 50
mg/day, less than 60 mg/day, less than 70 mg/day, less than 80
mg/day, less than 90 mg/day, or less than 100 mg/day. In some
embodiments, CHR-3996 is administered at a dose of more than 1
mg/day, more than 2 mg/day, more than 5 mg/day, more than 10
mg/day, more than 15 mg/day, more than 20 mg/day, more than 25
mg/day, more than 30 mg/day, more than 35 mg/day, more than 40
mg/day, more than 45 mg/day, more than 50 mg/day, more than 60
mg/day, more than 70 mg/day, more than 80 mg/day, more than 90
mg/day, or more than 100 mg/day. In certain embodiments, CHR-3996
is administered at a dose of more than 30 mg/day and less than 50
mg/day. In some embodiments, CHR-3996 is administered at a dose of
more than 20 mg/day and less than 80 mg/day. In certain
embodiments, CHR-3996 is administered once a day (q.d.), twice a
day (b.id.), or thrice a day (t.i.d.). In some embodiments,
CHR-3996 is administered daily, once a week, twice a week, three
times a week, four times a week, or five times a week.
[0112] In some embodiments, a unit dose of a co-formulated HDAC
inhibitor and antiviral agent comprises less than 100 mg of the
HDAC inhibitor and less than 1000 mg of the antiviral agent. In
certain embodiments, the unit dose comprises less than 50 mg of the
HDAC inhibitor and less than 500 mg of the antiviral agent. In
other embodiments, the unit dose comprises less than 80 mg of the
HDAC inhibitor and less than 1500 mg of the antiviral agent. In
some embodiments, the unit dose comprises less than 50 mg of the
HDAC inhibitor CHR-3996 and less than 1000 mg of valganciclovir. In
some embodiments, the unit dose comprises about 20 mg of the HDAC
inhibitor CHR-3996 and about 450 mg of valganciclovir. In certain
embodiments, the unit dose comprises about 40 mg of the HDAC
inhibitor CHR-3996 and about 900 mg of valganciclovir. In some
embodiments, the antiviral agent is formulated as controlled
release.
[0113] In some embodiments, the HDAC inhibitor is ITF-2357. In
certain embodiments, ITF-2357 is administered at a dose of 100
mg/day. In some embodiments, ITF-2357 is administered at a dose of
about 1 mg/day, about 2 mg/day, about 5 mg/day, about 10 mg/day,
about 15 mg/day, about 20 mg/day, about 25 mg/day, about 30 mg/day,
about 35 mg/day, about 40 mg/day, about 45 mg/day, about 50 mg/day,
about 60 mg/day, about 70 mg/day, about 80 mg/day, about 90 mg/day,
about 100 mg/day, about 120 mg/day, about 125 mg/day, about 140
mg/day, about 150 mg/day, about 160 mg/day, about 175 mg/day, about
180 mg/day, about 200 mg/day, about 225 mg/day, about 250 mg/day,
about 275 mg/day, or about 300 mg/day. In certain embodiments,
ITF-2357 is administered at a dose of less than 1 mg/day, less than
2 mg/day, less than 5 mg/day, less than 10 mg/day, less than 15
mg/day, less than 20 mg/day, less than 25 mg/day, less than 30
mg/day, less than 35 mg/day, less than 40 mg/day, less than 45
mg/day, less than 50 mg/day, less than 60 mg/day, less than 70
mg/day, less than 80 mg/day, less than 90 mg/day, less than 100
mg/day, less than 120 mg/day, less than 125 mg/day, less than 140
mg/day, less than 150 mg/day, less than 160 mg/day, less than 175
mg/day, less than 180 mg/day, less than 200 mg/day, less than 225
mg/day, less than 250 mg/day, less than 275 mg/day, or less than
300 mg/day. In some embodiments, ITF-2357 is administered at a dose
of more than 1 mg/day, more than 2 mg/day, more than 5 mg/day, more
than 10 mg/day, more than 15 mg/day, more than 20 mg/day, more than
25 mg/day, more than 30 mg/day, more than 35 mg/day, more than 40
mg/day, more than 45 mg/day, more than 50 mg/day, more than 60
mg/day, more than 70 mg/day, more than 80 mg/day, more than 90
mg/day, more than 100 mg/day, more than 120 mg/day, more than 125
mg/day, more than 140 mg/day, more than 150 mg/day, more than 160
mg/day, more than 175 mg/day, more than 180 mg/day, more than 200
mg/day, more than 225 mg/day, more than 250 mg/day, more than 275
mg/day, or more than 300 mg/day. In certain embodiments, ITF-2357
is administered at a dose of more than 80 mg/day and less than 120
mg/day. In some embodiments, ITF-2357 is administered at a dose of
more than 40 mg/day and less than 120 mg/day. In certain
embodiments, ITF-2357 is administered at a dose of more than 50
mg/day and less than 240 mg/day. In some embodiments, ITF-2357 is
administered once a day (q.d.), twice a day (b.id.), or thrice a
day (t.i.d.). In some embodiments, ITF-2357 is administered daily,
once a week, twice a week, three times a week, four times a week,
or five times a week.
[0114] In some embodiments, the HDAC inhibitor is
JNJ-16241199/R306465. In certain embodiments, JNJ-16241199/R306465
is administered at a dose of 100 mg/day. In some embodiments,
JNJ-16241199/R306465 is administered at a dose of about 1 mg/day,
about 2 mg/day, about 5 mg/day, about 10 mg/day, about 15 mg/day,
about 20 mg/day, about 25 mg/day, about 30 mg/day, about 35 mg/day,
about 40 mg/day, about 45 mg/day, about 50 mg/day, about 60 mg/day,
about 70 mg/day, about 80 mg/day, about 90 mg/day, about 100
mg/day, about 120 mg/day, about 125 mg/day, about 140 mg/day, about
150 mg/day, about 160 mg/day, about 175 mg/day, about 180 mg/day,
about 200 mg/day, about 225 mg/day, about 250 mg/day, about 275
mg/day, or about 300 mg/day. In certain embodiments,
JNJ-16241199/R306465 is administered at a dose of less than 1
mg/day, less than 2 mg/day, less than 5 mg/day, less than 10
mg/day, less than 15 mg/day, less than 20 mg/day, less than 25
mg/day, less than 30 mg/day, less than 35 mg/day, less than 40
mg/day, less than 45 mg/day, less than 50 mg/day, less than 60
mg/day, less than 70 mg/day, less than 80 mg/day, less than 90
mg/day, less than 100 mg/day, less than 120 mg/day, less than 125
mg/day, less than 140 mg/day, less than 150 mg/day, less than 160
mg/day, less than 175 mg/day, less than 180 mg/day, less than 200
mg/day, less than 225 mg/day, less than 250 mg/day, less than 275
mg/day, or less than 300 mg/day. In some embodiments,
JNJ-16241199/R306465 is administered at a dose of more than 1
mg/day, more than 2 mg/day, more than 5 mg/day, more than 10
mg/day, more than 15 mg/day, more than 20 mg/day, more than 25
mg/day, more than 30 mg/day, more than 35 mg/day, more than 40
mg/day, more than 45 mg/day, more than 50 mg/day, more than 60
mg/day, more than 70 mg/day, more than 80 mg/day, more than 90
mg/day, more than 100 mg/day, more than 120 mg/day, more than 125
mg/day, more than 140 mg/day, more than 150 mg/day, more than 160
mg/day, more than 175 mg/day, more than 180 mg/day, more than 200
mg/day, more than 225 mg/day, more than 250 mg/day, more than 275
mg/day, or more than 300 mg/day. In certain embodiments,
JNJ-16241199/R306465 is administered at a dose of more than 80
mg/day and less than 120 mg/day. In some embodiments,
JNJ-16241199/R306465 is administered at a dose of more than 40
mg/day and less than 120 mg/day. In certain embodiments,
JNJ-16241199/R306465 is administered at a dose of more than 50
mg/day and less than 240 mg/day. In some embodiments,
JNJ-16241199/R306465 is administered once a day (q.d.), twice a day
(b.id.), or thrice a day (t.i.d.). In some embodiments,
JNJ-16241199/R306465 is administered daily, once a week, twice a
week, three times a week, four times a week, or five times a
week.
[0115] In some embodiments, the HDAC inhibitor is JNJ-26481585. In
certain embodiments, JNJ-26481585 is administered at a dose of 10
mg/day. In some embodiments, JNJ-26481585 is administered at a dose
of about 0.1 mg/day, about 0.2 mg/day, about 0.5 mg/day, about 1
mg/day, about 2 mg/day, about 5 mg/day, about 10 mg/day, about 15
mg/day, about 20 mg/day, about 25 mg/day, about 30 mg/day, about 35
mg/day, about 40 mg/day, about 45 mg/day, about 50 mg/day, about 60
mg/day, about 70 mg/day, about 80 mg/day, about 90 mg/day, or about
100 mg/day. In certain embodiments, JNJ-26481585 is administered at
a dose of less than 0.1 mg/day, less than 0.2 mg/day, less than 0.5
mg/day, less than 1 mg/day, less than 2 mg/day, less than 5 mg/day,
less than 10 mg/day, less than 15 mg/day, less than 20 mg/day, less
than 25 mg/day, less than 30 mg/day, less than 35 mg/day, less than
40 mg/day, less than 45 mg/day, less than 50 mg/day, less than 60
mg/day, less than 70 mg/day, less than 80 mg/day, less than 90
mg/day, or less than 100 mg/day. In some embodiments, JNJ-26481585
is administered at a dose of more than 0.1 mg/day, more than 0.2
mg/day, more than 0.5 mg/day, more than 1 mg/day, more than 2
mg/day, more than 5 mg/day, more than 10 mg/day, more than 15
mg/day, more than 20 mg/day, more than 25 mg/day, more than 30
mg/day, more than 35 mg/day, more than 40 mg/day, more than 45
mg/day, more than 50 mg/day, more than 60 mg/day, more than 70
mg/day, more than 80 mg/day, more than 90 mg/day, or more than 100
mg/day. In certain embodiments, JNJ-26481585 is administered at a
dose of more than 2 mg/day and less than 20 mg/day. In some
embodiments, JNJ-26481585 is administered at a dose of more than 5
mg/day and less than 30 mg/day. In certain embodiments,
JNJ-26481585 is administered once a day (q.d.), twice a day
(b.id.), or thrice a day (t.i.d.). In some embodiments,
JNJ-26481585 is administered daily, once a week, twice a week,
three times a week, four times a week, or five times a week.
[0116] In some embodiments, the HDAC inhibitor is MGCD103. In
certain embodiments, MGCD103 is administered at a dose of 45
mg/m.sup.2/day. In some embodiments, MGCD103 is administered at a
dose of about 1 mg/m.sup.2/day, about 2 mg/m.sup.2/day, about 5
mg/m.sup.2/day, about 10 mg/m.sup.2/day, about 15 mg/m.sup.2/day,
about 20 mg/m.sup.2/day, about 25 mg/m.sup.2/day, about 30
mg/m.sup.2/day, about 35 mg/m.sup.2/day, about 40 mg/m2/day, about
45 mg/m.sup.2/day, about 50 mg/m.sup.2/day, about 60
mg/m.sup.2/day, about 70 mg/m.sup.2/day, about 80 mg/m.sup.2/day,
about 90 mg/m.sup.2/day, or about 100 mg/m.sup.2/day. In certain
embodiments, MGCD103 is administered at a dose of less than 1
mg/m.sup.2/day, less than 2 mg/m.sup.2/day, less than 5
mg/m.sup.2/day, less than 10 mg/m.sup.2/day, less than 15
mg/m.sup.2/day, less than 20 mg/m.sup.2/day, less than 25
mg/m.sup.2/day, less than 30 mg/m.sup.2/day, less than 35
mg/m.sup.2/day, less than 40 mg/m.sup.2/day, less than 45
mg/m.sup.2/day, less than 50 mg/m.sup.2/day, less than 60
mg/m.sup.2/day, less than 70 mg/m.sup.2/day, less than 80
mg/m.sup.2/day, less than 90 mg/m.sup.2/day, or less than 100
mg/m.sup.2/day. In some embodiments, MGCD103 is administered at a
dose of more than 1 mg/m.sup.2/day, more than 2 mg/m.sup.2/day,
more than 5 mg/m.sup.2/day, more than 10 mg/m.sup.2/day, more than
15 mg/m.sup.2/day, more than 20 mg/m.sup.2/day, more than 25
mg/m.sup.2/day, more than 30 mg/m.sup.2/day, more than 35
mg/m.sup.2/day, more than 40 mg/m.sup.2/day, more than 45
mg/m.sup.2/day, more than 50 mg/m.sup.2/day, more than 60
mg/m.sup.2/day, more than 70 mg/m.sup.2/day, more than 80
mg/m.sup.2/day, more than 90 mg/m.sup.2/day, or more than 100
mg/m.sup.2/day. In certain embodiments, MGCD103 is administered at
a dose of more than 30 mg/m.sup.2/day and less than 80
mg/m.sup.2/day. In some embodiments, MGCD103 is administered at a
dose of more than 45 mg/m.sup.2/day and less than 60
mg/m.sup.2/day. In certain embodiments, MGCD103 is administered
once a day (q.d.), twice a day (b.id.), or thrice a day (t.i.d.).
In some embodiments, MGCD103 is administered daily, once a week,
twice a week, three times a week, four times a week, or five times
a week.
[0117] In some embodiments, the HDAC inhibitor is MS-275. In
certain embodiments, MS-275 is administered at a dose of 4
mg/m.sup.2/day. In some embodiments, MS-275 is administered at a
dose of about 0.1 mg/m.sup.2/day, of about 0.2 mg/m.sup.2/day, of
about 0.5 mg/m.sup.2/day, of about 1 mg/m.sup.2/day, of about 2
mg/m.sup.2/day, of about 3 mg/m.sup.2/day, of about 4
mg/m.sup.2/day, about 5 mg/m.sup.2/day, of about 6 mg/m.sup.2/day,
of about 7 mg/m.sup.2/day, of about 8 mg/m.sup.2/day, of about 9
mg/m.sup.2/day, about 10 mg/m.sup.2/day, about 15 mg/m.sup.2/day,
about 20 mg/m.sup.2/day, about 25 mg/m.sup.2/day, about 30
mg/m.sup.2/day, about 35 mg/m.sup.2/day, about 40 mg/m2/day, about
45 mg/m.sup.2/day, about 50 mg/m.sup.2/day, about 60
mg/m.sup.2/day, about 70 mg/m.sup.2/day, about 80 mg/m.sup.2/day,
about 90 mg/m.sup.2/day, or about 100 mg/m.sup.2/day. In certain
embodiments, MS-275 is administered at a dose of less than 0.1
mg/m.sup.2/day, of less than 0.2 mg/m.sup.2/day, of less than 0.5
mg/m.sup.2/day, of less than 1 mg/m.sup.2/day, of less than 2
mg/m.sup.2/day, of less than 3 mg/m.sup.2/day, of less than 4
mg/m.sup.2/day, less than 5 mg/m.sup.2/day, of less than 6
mg/m.sup.2/day, of less than 7 mg/m.sup.2/day, of less than 8
mg/m.sup.2/day, of less than 9 mg/m.sup.2/day, less than 10
mg/m.sup.2/day, less than 15 mg/m.sup.2/day, less than 20
mg/m.sup.2/day, less than 25 mg/m.sup.2/day, less than 30
mg/m.sup.2/day, less than 35 mg/m.sup.2/day, less than 40
mg/m2/day, less than 45 mg/m.sup.2/day, less than 50
mg/m.sup.2/day, less than 60 mg/m.sup.2/day, less than 70
mg/m.sup.2/day, less than 80 mg/m.sup.2/day, less than 90
mg/m.sup.2/day, or less than 100 mg/m.sup.2/day. In some
embodiments, MS-275 is administered at a dose of more than 0.1
mg/m.sup.2/day, of more than 0.2 mg/m.sup.2/day, of more than 0.5
mg/m.sup.2/day, of more than 1 mg/m.sup.2/day, of more than 2
mg/m.sup.2/day, of more than 3 mg/m.sup.2/day, of more than 4
mg/m.sup.2/day, more than 5 mg/m.sup.2/day, of more than 6
mg/m.sup.2/day, of more than 7 mg/m.sup.2/day, of more than 8
mg/m.sup.2/day, of more than 9 mg/m.sup.2/day, more than 10
mg/m.sup.2/day, more than 15 mg/m.sup.2/day, more than 20
mg/m.sup.2/day, more than 25 mg/m.sup.2/day, more than 30
mg/m.sup.2/day, more than 35 mg/m.sup.2/day, more than 40
mg/m2/day, more than 45 mg/m.sup.2/day, more than 50
mg/m.sup.2/day, more than 60 mg/m.sup.2/day, more than 70
mg/m.sup.2/day, more than 80 mg/m.sup.2/day, more than 90
mg/m.sup.2/day, or more than 100 mg/m.sup.2/day. In certain
embodiments, MS-275 is administered at a dose of more than 2
mg/m.sup.2/day and less than 10 mg/m.sup.2/day. In some
embodiments, MS-275 is administered at a dose of more than 2
mg/m.sup.2/day and less than 40 mg/m.sup.2/day. In certain
embodiments, MS-275 is administered at a dose of more than 2
mg/m.sup.2/day and less than 6 mg/m.sup.2/day. In some embodiments,
MS-275 is administered at a dose of more than 6 mg/m.sup.2/day and
less than 8 mg/m.sup.2/day. In certain embodiments, MS-275 is
administered once a day (q.d.), twice a day (b.id.), or thrice a
day (t.i.d.). In some embodiments, MS-275 is administered daily,
once a week, twice a week, three times a week, four times a week,
or five times a week.
[0118] In some embodiments, the HDAC inhibitor is SB939. In certain
embodiments, SB939 is administered at a dose of 60 mg/day. In some
embodiments, SB939 is administered at a dose of about 1 mg/day,
about 2 mg/day, about 5 mg/day, about 10 mg/day, about 15 mg/day,
about 20 mg/day, about 25 mg/day, about 30 mg/day, about 35 mg/day,
about 40 mg/day, about 45 mg/day, about 50 mg/day, about 60 mg/day,
about 70 mg/day, about 80 mg/day, about 90 mg/day, about 100
mg/day, about 120 mg/day, about 125 mg/day, about 140 mg/day, about
150 mg/day, about 160 mg/day, about 175 mg/day, about 180 mg/day,
or about 200 mg/day. In certain embodiments, SB939 is administered
at a dose of less than 1 mg/day, less than 2 mg/day, less than 5
mg/day, less than 10 mg/day, less than 15 mg/day, less than 20
mg/day, less than 25 mg/day, less than 30 mg/day, less than 35
mg/day, less than 40 mg/day, less than 45 mg/day, less than 50
mg/day, less than 60 mg/day, less than 70 mg/day, less than 80
mg/day, less than 90 mg/day, less than 100 mg/day, less than 120
mg/day, less than 125 mg/day, less than 140 mg/day, less than 150
mg/day, less than 160 mg/day, less than 175 mg/day, less than 180
mg/day, or less than 200 mg/day. In some embodiments, SB939 is
administered at a dose of more than 1 mg/day, more than 2 mg/day,
more than 5 mg/day, more than 10 mg/day, more than 15 mg/day, more
than 20 mg/day, more than 25 mg/day, more than 30 mg/day, more than
35 mg/day, more than 40 mg/day, more than 45 mg/day, more than 50
mg/day, more than 60 mg/day, more than 70 mg/day, more than 80
mg/day, more than 90 mg/day, more than 100 mg/day, more than 120
mg/day, more than 125 mg/day, more than 140 mg/day, more than 150
mg/day, more than 160 mg/day, more than 175 mg/day, more than 180
mg/day, or more than 200 mg/day. In certain embodiments, SB939 is
administered at a dose of more than 30 mg/day and less than 70
mg/day. In some embodiments, SB939 is administered at a dose of
more than 10 mg/day and less than 90 mg/day. In certain
embodiments, SB939 is administered once a day (q.d.), twice a day
(b.id.), or thrice a day (t.i.d.). In some embodiments, SB939 is
administered daily, once a week, twice a week, three times a week,
four times a week, or five times a week.
[0119] In some embodiments, the HDAC inhibitor is romidepsin. In
certain embodiments, romidepsin is administered at a dose of 14
mg/m.sup.2/day. In some embodiments, romidepsin is administered at
a dose of about 0.1 mg/m.sup.2/day, of about 0.2 mg/m.sup.2/day, of
about 0.5 mg/m.sup.2/day, of about 1 mg/m.sup.2/day, of about 2
mg/m.sup.2/day, of about 3 mg/m.sup.2/day, of about 4
mg/m.sup.2/day, about 5 mg/m.sup.2/day, of about 6 mg/m.sup.2/day,
of about 7 mg/m.sup.2/day, of about 8 mg/m.sup.2/day, of about 9
mg/m.sup.2/day, about 10 mg/m.sup.2/day, about 11 mg/m.sup.2/day,
about 12 mg/m.sup.2/day, about 13 mg/m.sup.2/day, about 14
mg/m.sup.2/day, about 15 mg/m.sup.2/day, about 16 mg/m.sup.2/day,
about 17 mg/m.sup.2/day, about 18 mg/m.sup.2/day, about 19
mg/m.sup.2/day, about 20 mg/m.sup.2/day, about 25 mg/m.sup.2/day,
about 30 mg/m.sup.2/day, about 35 mg/m.sup.2/day, about 40
mg/m2/day, about 45 mg/m.sup.2/day, about 50 mg/m.sup.2/day, about
60 mg/m.sup.2/day, about 70 mg/m.sup.2/day, about 80
mg/m.sup.2/day, about 90 mg/m.sup.2/day, or about 100
mg/m.sup.2/day. In certain embodiments, romidepsin is administered
at a dose of less than 0.1 mg/m.sup.2/day, of less than 0.2
mg/m.sup.2/day, of less than 0.5 mg/m.sup.2/day, of less than 1
mg/m.sup.2/day, of less than 2 mg/m.sup.2/day, of less than 3
mg/m.sup.2/day, of less than 4 mg/m.sup.2/day, less than 5
mg/m.sup.2/day, of less than 6 mg/m.sup.2/day, of less than 7
mg/m.sup.2/day, of less than 8 mg/m.sup.2/day, of less than 9
mg/m.sup.2/day, less than 10 mg/m.sup.2/day, less than 11
mg/m.sup.2/day, less than 12 mg/m.sup.2/day, less than 13
mg/m.sup.2/day, less than 14 mg/m.sup.2/day, less than 15
mg/m.sup.2/day, less than 16 mg/m.sup.2/day, less than 17
mg/m.sup.2/day, less than 18 mg/m.sup.2/day, less than 19
mg/m.sup.2/day, less than 20 mg/m.sup.2/day, less than 25
mg/m.sup.2/day, less than 30 mg/m.sup.2/day, less than 35
mg/m.sup.2/day, less than 40 mg/m2/day, less than 45
mg/m.sup.2/day, less than 50 mg/m.sup.2/day, less than 60
mg/m.sup.2/day, less than 70 mg/m.sup.2/day, less than 80
mg/m.sup.2/day, less than 90 mg/m.sup.2/day, or less than 100
mg/m.sup.2/day. In some embodiments, romidepsin is administered at
a dose of more than 0.1 mg/m.sup.2/day, of more than 0.2
mg/m.sup.2/day, of more than 0.5 mg/m.sup.2/day, of more than 1
mg/m.sup.2/day, of more than 2 mg/m.sup.2/day, of more than 3
mg/m.sup.2/day, of more than 4 mg/m.sup.2/day, more than 5
mg/m.sup.2/day, of more than 6 mg/m.sup.2/day, of more than 7
mg/m.sup.2/day, of more than 8 mg/m.sup.2/day, of more than 9
mg/m.sup.2/day, more than 10 mg/m.sup.2/day, more than 11
mg/m.sup.2/day, more than 12 mg/m.sup.2/day, more than 13
mg/m.sup.2/day, more than 14 mg/m.sup.2/day, more than 15
mg/m.sup.2/day, more than 16 mg/m.sup.2/day, more than 17
mg/m.sup.2/day, more than 18 mg/m.sup.2/day, more than 19
mg/m.sup.2/day, more than 20 mg/m.sup.2/day, more than 25
mg/m.sup.2/day, more than 30 mg/m.sup.2/day, more than 35
mg/m.sup.2/day, more than 40 mg/m2/day, more than 45
mg/m.sup.2/day, more than 50 mg/m.sup.2/day, more than 60
mg/m.sup.2/day, more than 70 mg/m.sup.2/day, more than 80
mg/m.sup.2/day, more than 90 mg/m.sup.2/day, or more than 100
mg/m.sup.2/day. In certain embodiments, romidepsin is administered
at a dose of more than 13 mg/m.sup.2/day and less than 18
mg/m.sup.2/day. In some embodiments, romidepsin is administered at
a dose of more than 10 mg/m.sup.2/day and less than 20
mg/m.sup.2/day. In certain embodiments, romidepsin is administered
once a day (q.d.), twice a day (b.id.), or thrice a day (t.i.d.).
In some embodiments, romidepsin is administered daily, once a week,
twice a week, three times a week, four times a week, or five times
a week.
[0120] In some embodiments, the HDAC inhibitor is LBH589. In
certain embodiments, LBH589 is administered at a dose of 20 mg/day.
In some embodiments, LBH589 is administered at a dose of about 1
mg/day, about 2 mg/day, about 5 mg/day, about 10 mg/day, about 15
mg/day, about 20 mg/day, about 25 mg/day, about 30 mg/day, about 35
mg/day, about 40 mg/day, about 45 mg/day, about 50 mg/day, about 60
mg/day, about 70 mg/day, about 80 mg/day, about 90 mg/day, or about
100 mg/day. In certain embodiments, LBH589 is administered at a
dose of less than 1 mg/day, less than 2 mg/day, less than 5 mg/day,
less than 10 mg/day, less than 15 mg/day, less than 20 mg/day, less
than 25 mg/day, less than 30 mg/day, less than 35 mg/day, less than
40 mg/day, less than 45 mg/day, less than 50 mg/day, less than 60
mg/day, less than 70 mg/day, less than 80 mg/day, less than 90
mg/day, or less than 100 mg/day. In some embodiments, LBH589 is
administered at a dose of more than 1 mg/day, more than 2 mg/day,
more than 5 mg/day, more than 10 mg/day, more than 15 mg/day, more
than 20 mg/day, more than 25 mg/day, more than 30 mg/day, more than
35 mg/day, more than 40 mg/day, more than 45 mg/day, more than 50
mg/day, more than 60 mg/day, more than 70 mg/day, more than 80
mg/day, more than 90 mg/day, or more than 100 mg/day. In certain
embodiments, LBH589 is administered at a dose of more than 10
mg/day and less than 20 mg/day. In some embodiments, LBH589 is
administered at a dose of more than 5 mg/day and less than 30
mg/day. In certain embodiments, LBH589 is administered once a day
(q.d.), twice a day (b.id.), or thrice a day (t.i.d.). In some
embodiments, LBH589 is administered daily, once a week, twice a
week, three times a week, four times a week, or five times a
week.
[0121] In some embodiments, the HDAC inhibitor is PXD101. In
certain embodiments, PXD101 is administered at a dose of 1000
mg/m.sup.2/day. In some embodiments, PXD101 is administered at a
dose of about 10 mg/m.sup.2/day, about 15 mg/m.sup.2/day, about 20
mg/m.sup.2/day, about 50 mg/m.sup.2/day, about 75 mg/m.sup.2/day,
about 100 mg/m.sup.2/day, about 150 mg/m.sup.2/day, about 200
mg/m.sup.2/day, about 300 mg/m.sup.2/day, about 400 mg/m.sup.2/day,
about 500 mg/m.sup.2/day, about 600 mg/m.sup.2/day, about 700
mg/m.sup.2/day, about 800 mg/m.sup.2/day, about 900 mg/m.sup.2/day,
about 1000 mg/m.sup.2/day, about 1100 mg/m.sup.2/day, about 1200
mg/m.sup.2/day, about 1300 mg/m.sup.2/day, about 1400
mg/m.sup.2/day, about 1500 mg/m.sup.2/day, about 1750
mg/m.sup.2/day, or about 2000 mg/m.sup.2/day. In certain
embodiments, PXD101 is administered at a dose of less than 10
mg/m.sup.2/day, less than 15 mg/m.sup.2/day, less than 20
mg/m.sup.2/day, less than 50 mg/m.sup.2/day, less than 75
mg/m.sup.2/day, less than 100 mg/m.sup.2/day, less than 150
mg/m.sup.2/day, less than 200 mg/m.sup.2/day, less than 300
mg/m.sup.2/day, less than 400 mg/m.sup.2/day, less than 500
mg/m.sup.2/day, less than 600 mg/m.sup.2/day, less than 700
mg/m.sup.2/day, less than 800 mg/m.sup.2/day, less than 900
mg/m.sup.2/day, less than 1000 mg/m.sup.2/day, less than 1100
mg/m.sup.2/day, less than 1200 mg/m.sup.2/day, less than 1300
mg/m.sup.2/day, less than 1400 mg/m.sup.2/day, less than 1500
mg/m.sup.2/day, less than 1750 mg/m.sup.2/day, or less than 2000
mg/m.sup.2/day. In some embodiments, PXD101 is administered at a
dose of more than 10 mg/m.sup.2/day, more than 15 mg/m.sup.2/day,
more than 20 mg/m.sup.2/day, more than 50 mg/m.sup.2/day, more than
75 mg/m.sup.2/day, more than 100 mg/m.sup.2/day, more than 150
mg/m.sup.2/day, more than 200 mg/m.sup.2/day, more than 300
mg/m.sup.2/day, more than 400 mg/m.sup.2/day, more than 500
mg/m.sup.2/day, more than 600 mg/m.sup.2/day, more than 700
mg/m.sup.2/day, more than 800 mg/m.sup.2/day, more than 900
mg/m.sup.2/day, more than 1000 mg/m.sup.2/day, more than 1100
mg/m.sup.2/day, more than 1200 mg/m.sup.2/day, more than 1300
mg/m.sup.2/day, more than 1400 mg/m.sup.2/day, more than 1500
mg/m.sup.2/day, more than 1750 mg/m.sup.2/day, or more than 2000
mg/m.sup.2/day. In certain embodiments, PXD101 is administered at a
dose of more than 600 mg/m.sup.2/day and less than 1000
mg/m.sup.2/day. In some embodiments, PXD101 is administered at a
dose of more than 15 mg/m.sup.2/day and less than 1000
mg/m.sup.2/day. In certain embodiments, PXD101 is administered once
a day (q.d.), twice a day (b.id.), or thrice a day (t.i.d.). In
some embodiments, PXD101 is administered daily, once a week, twice
a week, three time a week, four times a week, or five times a
week.
[0122] In some embodiments, the HDAC inhibitor is vorinostat. In
certain embodiments, vorinostat is administered at a dose of 400
mg/day. In some embodiments, vorinostat is administered at a dose
of about 10 mg/day, about 20 mg/day, about 30 mg/day, about 40
mg/day, about 50 mg/day, about 60 mg/day, about 70 mg/day, about 80
mg/day, about 90 mg/day, about 100 mg/day, about 125 mg/day, about
150 mg/day, about 175 mg/day, about 200 mg/day, about 225 mg/day,
about 250 mg/day, about 275 mg/day, about 300 mg/day, about 325
mg/day, about 350 mg/day, about 375 mg/day, about 400 mg/day, about
450 mg/day, about 500 mg/day, about 550 mg/day, about 600 mg/day,
about 650 mg/day, about 700 mg/day, about 750 mg/day, about 800
mg/day, about 900 mg/day, or about 1000 mg/day. In certain
embodiments, vorinostat is administered at a dose of less than 10
mg/day, less than 20 mg/day, less than 30 mg/day, less than 40
mg/day, less than 50 mg/day, less than 60 mg/day, less than 70
mg/day, less than 80 mg/day, less than 90 mg/day, less than 100
mg/day, less than 125 mg/day, less than 150 mg/day, less than 175
mg/day, less than 200 mg/day, less than 225 mg/day, less than 250
mg/day, less than 275 mg/day, less than 300 mg/day, less than 325
mg/day, less than 350 mg/day, less than 375 mg/day, less than 400
mg/day, less than 450 mg/day, less than 500 mg/day, less than 550
mg/day, less than 600 mg/day, less than 650 mg/day, less than 700
mg/day, less than 750 mg/day, less than 800 mg/day, less than 900
mg/day, or less than 1000 mg/day. In some embodiments, vorinostat
is administered at a dose of more than 10 mg/day, more than 20
mg/day, more than 30 mg/day, more than 40 mg/day, more than 50
mg/day, more than 60 mg/day, more than 70 mg/day, more than 80
mg/day, more than 90 mg/day, more than 100 mg/day, more than 125
mg/day, more than 150 mg/day, more than 175 mg/day, more than 200
mg/day, more than 225 mg/day, more than 250 mg/day, more than 275
mg/day, more than 300 mg/day, more than 325 mg/day, more than 350
mg/day, more than 375 mg/day, more than 400 mg/day, more than 450
mg/day, more than 500 mg/day, more than 550 mg/day, more than 600
mg/day, more than 650 mg/day, more than 700 mg/day, more than 750
mg/day, more than 800 mg/day, more than 900 mg/day, or more than
1000 mg/day. In certain embodiments, vorinostat is administered at
a dose of more than 100 mg/day and less than 400 mg/day. In some
embodiments, vorinostat is administered at a dose of more than 100
mg/day and less than 500 mg/day. In certain embodiments, vorinostat
is administered once a day (q.d.), twice a day (b.id.), or thrice a
day (t.i.d.). In some embodiments, vorinostat is administered
daily, once a week, twice a week, three times a week, four times a
week, or five times a week.
[0123] In some embodiments, an HDAC inhibitor inhibits the growth
of virus-positive cells. In certain embodiments, the HDAC inhibitor
inhibits the growths of EBV-positive lymphoma cells. In some
embodiments, the HDAC inhibitor has growth inhibitory activity at a
concentration of about 100 .mu.M, about 90 .mu.M, about 80 .mu.M,
about 75 .mu.M, about 70 .mu.M, about 60 .mu.M, about 50 .mu.M,
about 40 .mu.M, about 30 .mu.M, about 25 .mu.M, about 20 .mu.M,
about 10 .mu.M, about 5 .mu.M, about 2 .mu.M, about 1 .mu.M, about
900 nM, about 800 nM, about 700 nM, about 600 nM, about 500 nM,
about 400 nM, about 300 nM, about 200 nM, about 100 nM, about 75
nM, about 50 nM, about 20 nM, or about 10 nM. In certain
embodiments, the HDAC inhibitor has growth inhibitory activity at a
concentration of less than 100 .mu.M, less than 90 .mu.M, less than
80 .mu.M, less than 75 .mu.M, less than 70 .mu.M, less than 60
.mu.M, less than 50 .mu.M, less than 40 .mu.M, less than 30 .mu.M,
less than 25 .mu.M, less than 20 .mu.M, less than 10 .mu.M, less
than 5 .mu.M, less than 2 .mu.M, less than 1 .mu.M, less than 900
nM, less than 800 nM, less than 700 nM, less than 600 nM, less than
500 nM, less than 400 nM, less than 300 nM, less than 200 nM, less
than 100 nM, less than 75 nM, less than 50 nM, less than 20 nM, or
less than 10 nM. In some embodiments, the HDAC inhibitor has growth
inhibitory activity at a concentration of more than 100 .mu.M, more
than 90 .mu.M, more than 80 .mu.M, more than 75 .mu.M, more than 70
.mu.M, more than 60 .mu.M, more than 50 .mu.M, more than 40 .mu.M,
more than 30 .mu.M, more than 25 .mu.M, more than 20 .mu.M, more
than 10 .mu.M, more than 5 .mu.M, more than 2 .mu.M, more than 1
.mu.M, more than 900 nM, more than 800 nM, more than 700 nM, more
than 600 nM, more than 500 nM, more than 400 nM, more than 300 nM,
more than 200 nM, more than 100 nM, more than 75 nM, more than 50
nM, more than 20 nM, or more than 10 nM. In certain embodiments,
the HDAC inhibitor has growth inhibitory activity at more than 50
nM and less than 100 nM. In some embodiments, the HDAC inhibitor
has growth inhibitory activity at more than 200 nM and less than
500 nM. In certain embodiments, the HDAC inhibitor has growth
inhibitory activity at more than 100 nM and less than 200 nM.
Induced Genes Including Viral-Associated Genes
[0124] Inducing agents (agents that induce expression) may act
directly on the viral genome or indirectly through a cellular
factor required for viral expression. For example, viral gene
expression can be regulated through the regulation of the
expression of viral transcription factors such as ZTA, RTA, tat,
and tax, cellular transcription factors such as AP-1, AP-2, Sp1,
NF-.kappa.B, and other transcriptional activators and/or repressors
(factors), co-activators and co-repressors, histone acetylators and
deacetylators, DNA methylases and demethylases, oncogenes or
proto-oncogenes, or protein kinase C. These proteins act to
regulate and thereby control expression of specific viral and/or
other cellular genetic elements. According to the methods of the
invention, control over their expression can lead to control over
the infection. Other gene products, both viral and cellular in
origin, whose expression can be regulated with inducing agents
include proteases, polymerases, reverse transcriptases,
cell-surface receptors, major histocompatibility antigens, growth
factors, and combination of these products.
[0125] Additional genes whose expression or transcriptional
regulation are altered in the presence of butyric acid include the
oncogenes myc, ras, myb, abl and src. The activities of these gene
products, as well as the activities of other oncogenes, are
described in Slamon, J. D., et al. 1984 Science 224:256-62.
Anti-proliferative activity also includes the ability to repress
tumor angiogenesis through the blockade of angiogenesis factor
activity, production or release, transcriptional regulation, or the
ability to modulate transcription of genes under angiogenesis or
growth factor or holinonal control. Either would be an effective
therapy, particularly against both prostatic neoplasia and breast
carcinomas. Further activities that effect transcription and/or
cellular differentiation include increased intracellular cAMP
levels, inhibition of histone acetylation, and inhibition of
genomic methylation. Each of these activities is directly related
to gene expression, and increased expression can sensitize infected
cells to a specific anti-viral agent.
[0126] In some embodiments, inducing agents include arginine
butyrate and/or other histone deacetylase inhibitors. Arginine
butyrate induces EBV-TK activity in EBV-immortalized B-cells and
patient-derived tumor cells. As latently-infected B-cells do not
express TK, exposure of these cells to agents like arginine
butyrate and/or other HDAC inhibitors results in a induction of
lytic replication and TK expression. This expression of a viral
gene can be used as a point for attack by anti-viral agents,
allowing for treatment of latent infections.
[0127] In other embodiments, inducing agents include HDAC
inhibitors that induce EBV-PK activity (also known BGLF4) in EBV
infected tumors. Expression of EBV-PK/BGLF4 sensitizes a cell to an
antiviral agent. In certain instances, HDAC inhibitors induce
EBV-PK. In some instances, HDAC inhibitors induce EBV-TK and/or
EBV-PK.
[0128] Preliminary in vitro studies according to the invention
demonstrate that induction of EBV-TK activity in EBV-immortalized
B-cells and patient-derived tumor cells using these drugs is
possible, and that these previously resistant cells are rendered
susceptible to ganciclovir therapy. Treatment of patients with
viral-associated tumors such as EBV with inducing agents such as
arginine butyrate, to induce the expression of EBV-TK/EBV-PK, and
GCV, to eliminate EBV-TK/EBV-PK expressing tumor cells, is an
effective, non-toxic therapy. This therapeutic regimen does not
depend on the associated viral genome being the cause of the tumor.
Without wishing to be bound by theory, it is believed that just the
presence of the EBV genome in latent form would make the tumor
susceptible to this combination protocol.
[0129] Butyrate-associated induction of genes has been
characterized for various cell types, and the genes are
consistently in the class of differentiation markers of a cell. For
example, in colon cancer cell lines, morphologic changes observed
in the presence of butyrate correlate with increased expression of
alkaline phosphatase, plasminogen activator, and CEA, all markers
of differentiation. Hepatoma cell lines increase expression of
alpha fetoprotein. Breast cancer cell lines express milk-related
glycoproteins, epithelial membrane antigens, and increased lipid
deposition. Sodium butyrate can also induce expression of cellular
proteins associated with converting basal keratinacytes into
committed epithelial cells.
[0130] Alteration of expression of certain transcription factors
may affect regulation of gene expression and regulation of the cell
cycle. In the breast cancer cell line MCF-7, butyrate induces a
block in cellular proliferation that is associated with decreased
expression of estrogen and prolactin hormone receptor mRNA
expression, thus blocking the potential growth stimulation by
estrogen and prolactin. These effects are associated with increased
expression of the EGF receptor. Butyrate also has been shown to
induce down-regulation of c-myc and p53 mRNA and to up-regulate
expression of the c-fos transcription factor. In mouse fibroblasts,
butyrate will block the cell cycle in the G.sub.1 phase. When these
cells are stimulated to proliferate with serum, TPA, or insulin,
the immediate-early response transcription factors c-myc and c-jun
are unregulated. However, the late G.sub.1 phase downstream gene
marker cdc-2 mRNA is not expressed, and cells are prevented from
entering S phase.
[0131] The particular combination of inducing agent with anti-viral
agent that is most effective against a specific disorder can be
determined by one of ordinary skill in the art from empirical
testing and, preferably, from a knowledge of each agent's mechanism
of action. Three such examples are as follows. First, many of the
RNA viruses such as HIV and other retroviruses require a reverse
transcriptase to transcribe their genome into DNA. A few of the
agents that induce expression or activity of retroviruses and their
encoded genes, such as, for example, reverse transcriptase, are
known to those of ordinary skill in the art. Anti-viral agents such
as nucleoside analogs can be administered to the patient. Those
substrate analogs will be specifically recognized by the reverse
transcriptase that, when incorporated into the infected-cell
genome, prevent viral replication and may also result in cell
death. Second, many viruses require an active protease to assemble
virus capsids to be packaged with viral genome. Protease inhibitors
or proteases that alter cleavage patterns so that packaging cannot
occur can be specifically targeted with an anti-viral agent that
comprises an amino acid analog or toxic conjugate. Third, arginine
butyrate and isobutyramide enhance expression of viral thymidine
kinase and other viral protein kinases in EBV-infected lymphocytes.
Ganciclovir or famcyclovir, in the presence of the viral thymidine
kinase or other viral kinases, destroys the infected cell.
Treatment of infected cells with both agents, according to the
invention, will selectively destroy EBV virus-infected cells. In
another aspect, of infected cells with both agents, according to
the invention, will selectively disable or disrupt the viral
activity within the cells in vivo.
[0132] In some embodiments, an inducing agent induces viral gene
expression by more than 4 fold after 24 h of treatment. In certain
embodiments, an HDAC inhibitor induces TK or EBV-PK expression by
more than 4 fold after 24 h of treatment. In some embodiments, an
HDAC inhibitor induces viral gene expression after about 48 h,
about 36 h, about 24 h, about 18 h, about 12 h, about 8 h, about 6
h, about 4 h, about 3 h, about 2 h, about 1 h, or about 30 minutes.
In certain embodiments, an HDAC inhibitor induces viral gene
expression in less than 48 h, less than 36 h, less than 24 h, less
than 18 h, less than 12 h, less than 8 h, less than 6 h, less than
4 h, less than 3 h, less than 2 h, less than 1 h, or less than 30
minutes. In some embodiments, an HDAC inhibitor induces viral gene
expression in more than 48 h, more than 36 h, more than 24 h, more
than 18 h, more than 12 h, more than 8 h, more than 6 h, more than
4 h, more than 3 h, more than 2 h, more than 1 h, or more than 30
minutes. In certain embodiments, an HDAC inhibitor induces viral
gene expression after more than 30 minutes and less than 24 h.
[0133] In certain embodiments, an inducing agent is capable of
inducing gene expression at a concentration of less than 500 nM. In
some embodiments, the inducing agent is an HDAC inhibitor. In
certain embodiments, the inducing agent is capable of inducing TK
or EBV-PK expression. In certain embodiments, an inducing agent is
capable of inducing gene expression at a concentration of about 100
.mu.M, about 90 .mu.M, about 80 .mu.M, about 75 .mu.M, about 70
.mu.M, about 60 .mu.M, about 50 .mu.M, about 40 .mu.M, about 30
.mu.M, about 25 .mu.M, about 20 .mu.M, about 10 .mu.M, about 5
.mu.M, about 2 .mu.M, about 1 .mu.M, about 900 nM, about 800 nM,
about 700 nM, about 600 nM, about 500 nM, about 400 nM, about 300
nM, about 200 nM, about 100 nM, about 75 nM, about 50 nM, about 20
nM, or about 10 nM. In some embodiments, an inducing agent is
capable of inducing gene expression at a concentration of less than
100 .mu.M, less than 90 .mu.M, less than 80 .mu.M, less than 75
.mu.M, less than 70 .mu.M, less than 60 .mu.M, less than 50 .mu.M,
less than 40 .mu.M, less than 30 .mu.M, less than 25 .mu.M, less
than 20 .mu.M, less than 10 .mu.M, less than 5 .mu.M, less than 2
.mu.M, less than 1 .mu.M, less than 900 nM, less than 800 nM, less
than 700 nM, less than 600 nM, less than 500 nM, less than 400 nM,
less than 300 nM, less than 200 nM, less than 100 nM, less than 75
nM, less than 50 nM, less than 20 nM, or less than 10 nM. In
certain embodiments, an inducing agent is capable of inducing gene
expression at a concentration of more than 100 .mu.M, more than 90
.mu.M, more than 80 .mu.M, more than 75 .mu.M, more than 70 .mu.M,
more than 60 .mu.M, more than 50 .mu.M, more than 40 .mu.M, more
than 30 .mu.M, more than 25 .mu.M, more than 20 .mu.M, more than 10
.mu.M, more than 5 .mu.M, more than 2 .mu.M, more than 1 .mu.M,
more than 900 nM, more than 800 nM, more than 700 nM, more than 600
nM, more than 500 nM, more than 400 nM, more than 300 nM, more than
200 nM, more than 100 nM, more than 75 nM, more than 50 nM, more
than 20 nM, or more than 10 nM. In some embodiments, an inducing
agent is capable of inducing gene expression at a concentration
more than 50 nM and less than 100 nM. In certain embodiments, an
inducing agent is capable of inducing gene expression at a
concentration of more than 200 nM and less than 500 nM. In some
embodiments, an inducing agent is capable of inducing gene
expression at more than 100 nM and less than 200 nM
[0134] In some embodiments, an HDAC inhibitor induces viral gene
expression after more than 1 h and less than 6 h. In certain
embodiments, an HDAC inhibitor induces viral gene expression about
2 fold, about 3 fold, about 4 fold, about 5 fold, about 6 fold,
about 7 fold, about 8 fold, about 9 fold, about 10 fold, about 12
fold, about 15 fold, about 20 fold, about 25 fold, about 30 fold,
about 35 fold, about 40 fold, about 45 fold, or about 50 fold. In
some embodiments, an HDAC inhibitor induces viral gene expression
less than 2 fold, less than 3 fold, less than 4 fold, less than 5
fold, less than 6 fold, less than 7 fold, less than 8 fold, less
than 9 fold, less than 10 fold, less than 12 fold, less than 15
fold, less than 20 fold, less than 25 fold, less than 30 fold, less
than 35 fold, less than 40 fold, less than 45 fold, or less than 50
fold. In certain embodiments, an HDAC inhibitor induces viral gene
expression more than 2 fold, more than 3 fold, more than 4 fold,
more than 5 fold, more than 6 fold, more than 7 fold, more than 8
fold, more than 9 fold, more than 10 fold, more than 12 fold, more
than 15 fold, more than 20 fold, more than 25 fold, more than 30
fold, more than 35 fold, more than 40 fold, more than 45 fold, or
more than 50 fold. In some embodiments, an HDAC inhibitor induces
viral gene expression more than 2 fold and less than 50 fold. In
certain embodiments, an HDAC inhibitor induces viral gene
expression more than 5 fold and less than 40 fold.
Antiviral Agents
[0135] Anti-viral agents that can be used in the compositions and
methods of the provided invention can include, for example,
substrates and substrate analogs, inhibitors and other agents that
severely impair, debilitate or otherwise destroy virus-infected
cells. Substrate analogs include amino acid and nucleoside analogs.
Substrates can be conjugated with toxins or other viricidal
substances Inhibitors include integrase inhibitors, protease
inhibitors, polymerase inhibitors and transcriptase inhibitors such
as reverse transcriptase inhibitors.
[0136] Antiviral agents that can be used in the compositions and
methods of the provided invention can include, for example,
ganciclovir, valganciclovir, oseltamivir (Tamiflu.TM.), zanamivir
(Relenza.TM.) abacavir, aciclovir, acyclovir, adefovir, amantadine,
amprenavir, ampligen, arbidol, atazanavir, atripla, boceprevir,
cidofovir, combivir, darunavir, delavirdine, didanosine, docosanol,
edoxudine, efavirenz, emtricitabine, enfuvirtide, entecavir,
famciclovir, fomivirsen, fosamprenavir, foscarnet, fosfonet, fusion
inhibitors (e.g., enfuvirtide), ibacitabine, imunovir, idoxuridine,
imiquimod, indinavir, inosine, integrase inhibitor, interferon type
III, interferon type II, interferon type I, interferon, lamivudine,
lopinavir, loviride, maraviroc, moroxydine, nelfinavir, nevirapine,
nexavir, nucleoside analogues, peginterferon alfa-2a, penciclovir,
peramivir, pleconaril, podophyllotoxin, protease inhibitor,
raltegravir, reverse transcriptase inhibitor, ribavirin,
rimantadine, ritonavir, pyrimidine antiviral, saquinavir,
stavudine, synergistic enhancer (antiretroviral), tenofovir,
tenofovir disoproxil, tipranavir, trifluridine, trizivir,
tromantadine, truvada, valaciclovir (Valtrex.TM.), vicriviroc,
vidarabine, viramidine, zalcitabine, and zidovudine.
[0137] In a specific embodiment, the antiviral agent is acyclovir,
ganciclovir, or valganciclovir.
[0138] In some embodiments, the antiviral agent is a nucleoside.
Examples of nucleoside analogs include acyclovir (ACV), ganciclovir
(GCV), valganciclovir, famciclovir, foscarnet, ribavirin,
zalcitabine (ddC), zidovudine (AZT), stavudine (D4T), lamivudine
(3TC), didanosine (ddI), cytarabine, dideoxyadenosine, edoxudine,
floxuridine, idozuridine, inosine pranobex,
2'-deoxy-5-(methylamino)uridine, trifluridine and vidarabine.
Examples of a few protease inhibitors that show particular promise
in human therapy include saquinivir, ritonavir and indinavir. Other
anti-viral agents include interferons (e.g. .alpha.-, .beta.-,
.gamma.-interferon), cytokines such as tumor necrosis factor (TNF)
or interleukins, cell receptors and growth factor antagonists,
which can be purified or recombinantly produced.
[0139] In some embodiments, the antiviral agent is administered at
a dose of less than 3000 mg/day. In some embodiments, the antiviral
agent is administered at a dose of about 10 mg/day, about 20
mg/day, about 50 mg/day, about 100 mg/day, about 150 mg/day, about
200 mg/day, about 250 mg/day, about 300 mg/day, about 350 mg/day,
about 400 mg/day, about 450 mg/day, about 500 mg/day, about 600
mg/day, about 700 mg/day, about 800 mg/day, about 900 mg/day, about
1000 mg/day, about 1200 mg/day, about 1250 mg/day, about 1400
mg/day, about 1500 mg/day, about 1600 mg/day, about 1750 mg/day,
about 1800 mg/day, about 1900 mg/day, about 2000 mg/day, about 2250
mg/day, about 2500 mg/day, about 2750 mg/day, about 3000 mg/day,
about 3250 mg/day, about 3500 mg/day, about 3750 mg/day, about 4000
mg/day, about 4250 mg/day, about 4500 mg/day, about 4750 mg/day, or
about 5000 mg/day. In certain embodiments, the antiviral agent is
administered at a dose of less than 10 mg/day, less than 20 mg/day,
less than 50 mg/day, less than 100 mg/day, less than 150 mg/day,
less than 200 mg/day, less than 250 mg/day, less than 300 mg/day,
less than 350 mg/day, less than 400 mg/day, less than 450 mg/day,
less than 500 mg/day, less than 600 mg/day, less than 700 mg/day,
less than 800 mg/day, less than 900 mg/day, less than 1000 mg/day,
less than 1200 mg/day, less than 1250 mg/day, less than 1400
mg/day, less than 1500 mg/day, less than 1600 mg/day, less than
1750 mg/day, less than 1800 mg/day, less than 1900 mg/day, less
than 2000 mg/day, less than 2250 mg/day, less than 2500 mg/day,
less than 2750 mg/day, less than 3000 mg/day, less than 3250
mg/day, less than 3500 mg/day, less than 3750 mg/day, less than
4000 mg/day, less than 4250 mg/day, less than 4500 mg/day, less
than 4750 mg/day, or less than 5000 mg/day. In some embodiments,
the antiviral agent is administered at a dose of more than 10
mg/day, more than 20 mg/day, more than 50 mg/day, more than 100
mg/day, more than 150 mg/day, more than 200 mg/day, more than 250
mg/day, more than 300 mg/day, more than 350 mg/day, more than 400
mg/day, more than 450 mg/day, more than 500 mg/day, more than 600
mg/day, more than 700 mg/day, more than 800 mg/day, more than 900
mg/day, more than 1000 mg/day, more than 1200 mg/day, more than
1250 mg/day, more than 1400 mg/day, more than 1500 mg/day, more
than 1600 mg/day, more than 1750 mg/day, more than 1800 mg/day,
more than 1900 mg/day, more than 2000 mg/day, more than 2250
mg/day, more than 2500 mg/day, more than 2750 mg/day, more than
3000 mg/day, more than 3250 mg/day, more than 3500 mg/day, more
than 3750 mg/day, more than 4000 mg/day, more than 4250 mg/day,
more than 4500 mg/day, more than 4750 mg/day, or more than 5000
mg/day. In certain embodiments, the antiviral agent is administered
at a dose of more than 10 mg/day and less than 5000 mg/day. In some
embodiments, the antiviral agent is administered at a dose of more
than 200 mg/day and less than 1000 mg/day. In certain embodiments,
the antiviral agent is administered once a day (q.d.), twice a day
(b.id.), or thrice a day (t.i.d.). In some embodiments, the
antiviral agent is administered daily, once a week, twice a week,
three times a week, four times a week, or five times a week.
[0140] In certain embodiments, the antiviral agent is ganciclovir.
In some embodiments, ganciclovir is administered at a total daily
dose of 3000 mg/day. In certain embodiments, ganciclovir is
administered at a dose of 1000 mg three times a day. In some
embodiments, ganciclovir is administered at a dose of about 100
mg/day, about 250 mg/day, about 500 mg/day, about 750 mg/day, about
1000 mg/day, about 1500 mg/day, about 2000 mg/day, about 2500
mg/day, about 3000 mg/day, about 3500 mg/day, or about 4000 mg/day.
In certain embodiments, ganciclovir is administered at a dose of
less than 100 mg/day, less than 250 mg/day, less than 500 mg/day,
less than 750 mg/day, less than 1000 mg/day, less than 1500 mg/day,
less than 2000 mg/day, less than 2500 mg/day, less than 3000
mg/day, less than 3500 mg/day, or less than 4000 mg/day. In some
embodiments, ganciclovir is administered at a dose of more than 100
mg/day, more than 250 mg/day, more than 500 mg/day, more than 750
mg/day, more than 1000 mg/day, more than 1500 mg/day, more than
2000 mg/day, more than 2500 mg/day, more than 3000 mg/day, more
than 3500 mg/day, or more than 4000 mg/day. In certain embodiments,
ganciclovir is administered at a dose of more than 500 mg/day and
less 4000 mg/day. In some embodiments, ganciclovir is administered
at a dose of more than 1000 mg/day and less than 3000 mg/day. In
some embodiments, ganciclovir is administered once a day, twice a
day, or three times a day. In certain embodiments, ganciclovir is
administered once a week, twice a week, three times a week, four
times a week, five times a week, or daily.
[0141] In some embodiments, the antiviral agent is valganciclovir.
In certain embodiments, valganciclovir is administered at a total
daily dose of 900 mg/day. In some embodiments, valganciclovir is
administered at a dose of 900 mg once a day. In certain
embodiments, valganciclovir is administered at a total daily dose
of 1800 mg/day. In some embodiments, valganciclovir is administered
at a dose of 900 mg twice a day.
[0142] In some embodiments, valganciclovir is administered at a
dose of about 100 mg/day, about 200 mg/day, about 300 mg/day, about
400 mg/day, about 500 mg/day, about 600 mg/day, about 700 mg/day,
about 800 mg/day, about 900 mg/day, about 1000 mg/day, about 1100
mg/day, about 1200 mg/day, about 1300 mg/day, about 1400 mg/day,
about 1500 mg/day, about 1600 mg/day, about 1700 mg/day, about 1800
mg/day, about 1900 mg/day, or about 2000 mg/day. In certain
embodiments, valganciclovir is administered at a dose of less than
100 mg/day, less than 200 mg/day, less than 300 mg/day, less than
400 mg/day, less than 500 mg/day, less than 600 mg/day, less than
700 mg/day, less than 800 mg/day, less than 900 mg/day, less than
1000 mg/day, less than 1100 mg/day, less than 1200 mg/day, less
than 1300 mg/day, less than 1400 mg/day, less than 1500 mg/day,
less than 1600 mg/day, less than 1700 mg/day, less than 1800
mg/day, less than 1900 mg/day, or less than 2000 mg/day. In some
embodiments, valganciclovir is administered at a dose of more than
100 mg/day, more than 200 mg/day, more than 300 mg/day, more than
400 mg/day, more than 500 mg/day, more than 600 mg/day, more than
700 mg/day, more than 800 mg/day, more than 900 mg/day, more than
1000 mg/day, more than 1100 mg/day, more than 1200 mg/day, more
than 1300 mg/day, more than 1400 mg/day, more than 1500 mg/day,
more than 1600 mg/day, more than 1700 mg/day, more than 1800
mg/day, more than 1900 mg/day, or more than 2000 mg/day. In certain
embodiments, valganciclovir is administered at a dose of more than
100 mg/day and less 2000 mg/day. In some embodiments,
valganciclovir is administered at a dose of more than 500 mg/day
and less than 1500 mg/day. In some embodiments, valganciclovir is
administered once a day, twice a day, or three times a day. In
certain embodiments, valganciclovir is administered once a week,
twice a week, three time a week, four times a week, five times a
week, or daily.
[0143] In a specific embodiment, the antiviral agent is not a heat
shock protein inhibitor, an immunosuppressant, an antibiotic, a
glucocorticoid, a non-steroidal anti-inflammatory drug, a
Cox-2-specific inhibitor or a TNF-.alpha. binding protein. In a
related embodiment, the antiviral agent is not a Hsp90 inhibitor,
tacrolimus, cyclosporin, rapamycin (sirolimus), methotrexate,
cyclophosphamide, azathioprine, mercaptopurine, mycophenolate,
FTY720, levofloxacin, amoxycillin, prednisone, cortisone acetate,
prednisolone, methylprednisolone, dexamethasone, betamethasone,
triamcinolone, beclometasone, fludrocortisone acetate,
deoxycorticosterone acetate, aldosterone, salicylates, arylalkanoic
acids, a 2-arylpropionic acid, a N-arylanthranilic acid, an oxicam,
a coxib, a sulphonanilide, valdecoxib, celecoxib, rofecoxib,
leflunomide, gold thioglucose, gold thiomalate, aurofin,
sulfasalazine, hydroxychloroquinine, minocycline, infliximab,
etanercept, adalimumab, abatacept, anakinra, interferon-.beta.,
interferon-.gamma., interleukin-2, an allergy vaccine, an
antihistamine, an antileukotriene, a beta-agonist, theophylline, or
an anticholinergic.
Additional Agents
[0144] The methods of the provided invention can comprise
administering to a subject a viral inducing agent, and antiviral
agent, and one or more additional active agents. The additional
agent can be selected based on the type of viral, virally-induced,
or inflammatory condition the subject has or is suspected of
having. The additional agent can comprise, for example, another
antiviral agent, another viral inducing agent, a vaccine, or an
anticancer agent. For example, a subject with multiple sclerosis
can be administered a viral inducing agent, an antiviral agent, and
a vaccine, for example, a vaccine comprising myelin basic protein.
In another example, a subject with diabetes can be administered a
viral inducing agent, an antiviral agent, and a vaccine, for
example, a vaccine comprising an antigen.
[0145] In some embodiments, an additional agent is a anticancer
agent. In certain embodiments, the anticancer agent is a
chemotherapeutic anticancer agent. Examples of chemotherapeutic
anticancer agents include, but are not limited to, nitrogen
mustards; alkyl sulfonates; ethylene imines; nitrosoureas;
epoxides; other alkylating agents; folic acid analogues; purine
analogs; pyrimidine analogs; vinca alkaloids; podophyllotoxin
derivatives; colchicine derivatives; taxanes; other plant alkaloids
and natural products; actinomycines; antracyclines; other cytotoxic
antibiotics; platinum compounds; methylhydrazines; sensitizers;
protein kinase inhibitors; other antineoplastic agents; estrogens;
progestogens; gonadotropin releasing hormone analogs;
anti-estrogens; anti-androgens; enzyme inhibitors; other hormone
antagonists; immunostimulants; immunosuppressants; calcineurin
inhibitors; and radiopharmaceuticals. In some embodiments, the
anticancer agent is a toxin, e.g. diphtheria toxin.
[0146] In certain embodiments, an additional agent is a
non-steroidal anti-inflammatory agent (NSAID). NSAID include, for
example, Aspirin (Anacin.TM., Ascriptin.TM., Bayer.TM.,
Bufferin.TM., Ecotrin.TM. Excedrin.TM.), Choline and magnesium
salicylates (CMT.TM., Tricosal.TM., Trilisate.TM.), choline
salicylate (Arthropan.TM.), celecoxib (Celebrex.TM.), diclofenac
potassium (Cataflam.TM.), diclofenac sodium (Voltaren.TM., Voltaren
XR.TM.), diclofenac sodium with misoprostol (Arthrotec.TM.),
diflunisal (Dolobid.TM.), etodolac (Lodine.TM., Lodine XL.TM.),
fenoprofen calcium (Nalfon.TM.), flurbiprofen (Ansaid.TM.),
ibuprofen (Advil.TM., Motrin.TM., Motrin IB.TM., Nuprin.TM.),
indomethacin (Indocin.TM. Indocin SR.TM.), ketoprofen (Actron.TM.,
Orudis.TM., Orudis KT.TM., Oruvail.TM.), magnesium salicylate
(Arthritab.TM., Bayer Select.TM., Doan's Pills.TM., Magan.TM.,
Mobidin.TM., Mobogesic.TM.), meclofenamate sodium (Meclomen.TM.),
mefenamic acid (Ponstel.TM.), meloxicam (Mobic.TM.), nabumetone
(Relafen.TM.) naproxen (Naprosyn.TM., Naprelan.TM.), naproxen
sodium (Aleve.TM., Anaprox.TM.), oxaprozin (Daypro.TM.), piroxicam
(Feldene.TM.), rofecoxib (Vioxx.TM.), salsalate (Amigesic.TM.,
Anaflex 750.TM., Disalcid.TM., Marthritic.TM., Mono-Gesic.TM.,
Salflex.TM., Salsitab.TM.), sodium salicylate (various generics),
sulindac (Clinoril.TM.), tolmetin sodium (Tolectin.TM.), valdecoxib
(Bextra.TM.).
[0147] In some embodiments, an additional agent is a lipid lowering
agent. In certain embodiments, the lipid lowering agent is a
statin. Examples of statins include, but are not limited to,
Advicor.RTM. (niacin extended-release/lovastatin), Altoprev.RTM.
(lovastatin extended-release), Caduet.RTM. (amlodipine and
atorvastatin), Crestor.RTM. (rosuvastatin), Lescol.RTM.
(fluvastatin), Lescol XL (fluvastatin extended-release),
Lipitor.RTM. (atorvastatin), Mevacor.RTM. (lovastatin),
Pravachol.RTM. (pravastatin), Simcor.RTM. (niacin
extended-release/simvastatin), Vytorin.RTM.
(ezetimibe/simvastatin), and Zocor.RTM. (simvastatin). A lipid
lowering agent can be administered to a subject that has or is
suspected of having atherosclerosis. For example, a subject with
cytomegalovirus induced atherosclerosis can be administered an
additional agent that can comprise atorvastatin, rosuvastatin,
lovastatin, simvastatin, or pravastatin.
[0148] In certain embodiments, an additional agent is an
immunosuppressive drug. Immunosuppressive drug, for example,
include glucocorticoids, antibodies, cytostatic agents, and drugs
that act on immunophilins. Glucocorticoids can include, for
example, prednisolone, prednisone, or methylprednisolone. A
cytostatic agent can include, for example, an agent that interferes
with nucleic acid synthesis, for example, folic acid, pyrimidine
analogs, and purine analogs. A folic acid analog that can be used
as an immunosuppressive drug is methotrexate, which can bind
dihydrofolate reductase and prevent the synthesis of
tetrahydrofolate. Another cytostatic agent is azathioprine, which
can be cleaved nonezymatically to form mercaptopurine, which can
act as a purine analogue. A cytostatic agent can include, for
example, an alkylating agent, including, for example,
cyclophosphamide, and a nitrosourea. A cytostatic agent can be a
platinum compound. Other cytostatic agents include, for example,
cytotoxic antibiotics, including dactinomycin, anthracylcines,
mitomycin C, bleomycin, and mithramycin. Examples of antibodies
that can be immunosuppressive agents include, for example,
heterologous polyclonal antibodies, for example, from rabbit or
horse. Other antibodies include monoclonal antibodies directed to
specific antigens e.g., T-cell receptor directed antibodies (e.g.,
OKT3, muromonab, which targets CD3), and IL-2 receptor directed
antibodies (e.g., targeting CD25). Drugs that can act on
immunophilins include, for example, cyclosporin, tacrolimus
(Prograf), Sirolimus (rapamycin, Rapamune). Other drugs that can
act as immunosuppressive drugs include, for example, mycophenolate
(mycophenolic acid), interferons, opioids, TNF binding proteins,
Fingolimod, myriocin, and ciclosporin.
[0149] The additional agent can be, for example, FK506, a
monoclonal antibody, an anti-T cell monoclonal antibody, an anti-B
cell monoclonal antibody, or a TNF inhibitor. The monoclonal
antibody can be an anti-B cell antibody. The anti-B cell antibody
can be anti-CD20. The TNF inhibitor can be infliximab
(Remicade.TM.), etanercept (Enbrel.TM.), adalimumab (Humira.TM.),
or an anti-IL-6 antibody.
[0150] A subject with an autoimmune condition can be administered a
viral inducing agent, an antiviral agent, and an additional agent,
where the additional agent comprises cyclosporine, azathiorprine,
methotrexate, cyclophosphamide, FK506, tacrolimus, a monoclonal
antibody, an anti-T cell monoclonal antibody, an anti-B cell
monoclonal antibody, an IL-2 receptor antibody, or a TNF
inhibitor.
[0151] The additional agent can be glatiramer (Copaxone.TM.),
Natalizumab (Tysabri.TM.), mitoxantrone (Novantrone.TM.),
cladribine, or Campath antibody. For example, a subject with
multiple sclerosis can be administered an additional agent that can
comprise glatiramer, mitoxantrone, natalizumab, cladribine, or
Campath antibody.
Types of Viruses and Virally-Induced Conditions
[0152] The methods and compositions provided herein can be used to
treat and/or prevent viral infections.
[0153] The virus causing the infection can be a member of the
herpes virus family, a human immunodeficiency virus, parvovirus, or
coxsackie virus. A member of the herpes virus family can be herpes
simplex virus, herpes genitalis virus, varicella zoster virus,
Epstein-Barr virus, human herpes virus 6, human herpes virus 8, or
cytomegalovirus. The subject can have coronary artery condition
associated with a cytomegalovirus or herpes simplex virus
infection. The subject can have autoimmune condition associated
with Epstein-Barr virus infection. The subject can have a lymphoma
or other cancer associated with Epstein-Barr virus infection.
[0154] In some embodiments, the viral or virally-induced condition
is caused by a retrovirus, such as HIV, HTLV1 and 2. In certain
embodiments, the viral or virally-induced condition is caused by a
DNA virus, such as a herpesvirus. In some embodiments, the
herpesvirus is an Epstein-Barr virus, cytomegalovirus, Herpes type
1 (oral herpes), herpes type 2, Kaposi's sarcoma virus (human
herpes virus 8), BK viruses, or hepatitis virus. In certain
embodiments, the virally-induced or virus-associated disease is a
cancer. In some embodiments, the virally-induced or
virus-associated cancer is a lymphoma, chronic lyphocytic leukemia,
nasopharyngeal carcinoma, gastric cancer, or Kaposi's sarcoma. In
other embodiments, the virally-induced or virus-associated disease
is an autoimmune disease. In certain embodiments, the autoimmune
disease is rheumatoid arthritis, systemic lupus erythematosus, or
multiple sclerosis.
[0155] The methods and compositions described herein can be used to
treat and/or prevent infections caused by any virus, including, for
example, Abelson leukemia virus, Abelson murine leukemia virus,
Abelson's virus, Acute laryngotracheobronchitis virus, Adelaide
River virus, Adeno associated virus group, Adenovirus, African
horse sickness virus, African swine fever virus, AIDS virus,
Aleutian mink condition parvovirus, Alpharetrovirus, Alphavirus,
ALV related virus, Amapari virus, Aphthovirus, Aquareovirus,
Arbovirus, Arbovirus C, arbovirus group A, arbovirus group B,
Arenavirus group, Argentine hemorrhagic fever virus, Argentine
hemorrhagic fever virus, Arterivirus, Astrovirus, Ateline
herpesvirus group, Aujezky's condition virus, Aura virus, Ausduk
condition virus, Australian bat lyssavirus, Aviadenovirus, avian
erythroblastosis virus, avian infectious bronchitis virus, avian
leukemia virus, avian leukosis virus, avian lymphomatosis virus,
avian myeloblastosis virus, avian paramyxovirus, avian
pneumoencephalitis virus, avian reticuloendotheliosis virus, avian
sarcoma virus, avian type C retrovirus group, Avihepadnavirus,
Avipoxvirus, B virus, B19 virus, Babanki virus, baboon herpesvirus,
baculovirus, Barmah Forest virus, Bebaru virus, Berrimah virus,
Betaretrovirus, Birnavirus, Bittner virus, BK virus, Black Creek
Canal virus, bluetongue virus, Bolivian hemorrhagic fever virus,
Boma condition virus, border condition of sheep virus, borna virus,
bovine alphaherpesvirus 1, bovine alphaherpesvirus 2, bovine
coronavirus, bovine ephemeral fever virus, bovine immunodeficiency
virus, bovine leukemia virus, bovine leukosis virus, bovine
mammillitis virus, bovine papillomavirus, bovine papular stomatitis
virus, bovine parvovirus, bovine syncytial virus, bovine type C
oncovirus, bovine viral diarrhea virus, Buggy Creek virus, bullet
shaped virus group, Bunyamwera virus supergroup, Bunyavirus,
Burkitt's lymphoma virus, Bwamba Fever, CA virus, Calicivirus,
California encephalitis virus, camelpox virus, canarypox virus,
canid herpesvirus, canine coronavirus, canine distemper virus,
canine herpesvirus, canine minute virus, canine parvovirus, Cano
Delgadito virus, caprine arthritis virus, caprine encephalitis
virus, Caprine Herpes Virus, Capripox virus, Cardiovirus, caviid
herpesvirus 1, Cercopithecid herpesvirus 1, cercopithecine
herpesvirus 1, Cercopithecine herpesvirus 2, Chandipura virus,
Changuinola virus, channel catfish virus, Charleville virus,
chickenpox virus, Chikungunya virus, chimpanzee herpesvirus, chub
reovirus, chum salmon virus, Cocal virus, Coho salmon reovirus,
coital exanthema virus, Colorado tick fever virus, Coltivirus,
Columbia SK virus, common cold virus, contagious ecthyma virus,
contagious pustular dermatitis virus, Coronavirus, Corriparta
virus, coryza virus, cowpox virus, coxsackie virus, CPV
(cytoplasmic polyhedrosis virus), cricket paralysis virus,
Crimean-Congo hemorrhagic fever virus, croup associated virus,
Cryptovirus, Cypovirus, Cytomegalovirus, cytomegalovirus group,
cytoplasmic polyhedrosis virus, deer papillomavirus,
deltaretrovirus, dengue virus, Densovirus, Dependovirus, Dhori
virus, diploma virus, Drosophila C virus, duck hepatitis B virus,
duck hepatitis virus 1, duck hepatitis virus 2, duovirus, Duvenhage
virus, Deformed wing virus DWV, eastern equine encephalitis virus,
eastern equine encephalomyelitis virus, EB virus, Ebola virus,
Ebola-like virus, echo virus, echovirus, echovirus 10, echovirus
28, echovirus 9, ectromelia virus, EEE virus, EIA virus, EIA virus,
encephalitis virus, encephalomyocarditis group virus,
encephalomyocarditis virus, Enterovirus, enzyme elevating virus,
enzyme elevating virus (LDH), epidemic hemorrhagic fever virus,
epizootic hemorrhagic condition virus, Epstein-Barr virus, equid
alphaherpesvirus 1, equid alphaherpesvirus 4, equid herpesvirus 2,
equine abortion virus, equine arteritis virus, equine encephalosis
virus, equine infectious anemia virus, equine morbillivirus, equine
rhinopneumonitis virus, equine rhinovirus, Eubenangu virus,
European elk papillomavirus, European swine fever virus, Everglades
virus, Eyach virus, felid herpesvirus 1, feline calicivirus, feline
fibrosarcoma virus, feline herpesvirus, feline immunodeficiency
virus, feline infectious peritonitis virus, feline leukemia/sarcoma
virus, feline leukemia virus, feline panleukopenia virus, feline
parvovirus, feline sarcoma virus, feline syncytial virus,
Filovirus, Flanders virus, Flavivirus, foot and mouth condition
virus, Fort Morgan virus, Four Corners hantavirus, fowl adenovirus
1, fowlpox virus, Friend virus, Gammaretrovirus, GB hepatitis
virus, GB virus, German measles virus, Getah virus, gibbon ape
leukemia virus, glandular fever virus, goatpox virus, golden
shinner virus, Gonometa virus, goose parvovirus, granulosis virus,
Gross' virus, ground squirrel hepatitis B virus, group A arbovirus,
Guanarito virus, guinea pig cytomegalovirus, guinea pig type C
virus, Hantaan virus, Hantavirus, hard clam reovirus, hare fibroma
virus, HCMV (human cytomegalovirus), hemadsorption virus 2,
hemagglutinating virus of Japan, hemorrhagic fever virus, hendra
virus, Henipaviruses, Hepadnavirus, hepatitis A virus, hepatitis B
virus group, hepatitis C virus, hepatitis D virus, hepatitis delta
virus, hepatitis E virus, hepatitis F virus, hepatitis G virus,
hepatitis nonA nonB virus, hepatitis virus, hepatitis virus
(nonhuman), hepatoencephalomyelitis reovirus 3, Hepatovirus, heron
hepatitis B virus, herpes B virus, herpes simplex virus, herpes
simplex virus 1, herpes simplex virus 2, herpesvirus, herpesvirus
7, Herpesvirus ateles, Herpesvirus hominis, Herpesvirus infection,
Herpesvirus saimiri, Herpesvirus suis, Herpesvirus varicellae,
Highlands J virus, Hirame rhabdovirus, hog cholera virus, human
adenovirus 2, human alphaherpesvirus 1, human alphaherpesvirus 2,
human alphaherpesvirus 3, human B lymphotropic virus, human
betaherpesvirus 5, human coronavirus, human cytomegalovirus group,
human foamy virus, human gammaherpesvirus 4, human gammaherpesvirus
6, human hepatitis A virus, human herpesvirus 1 group, human
herpesvirus 2 group, human herpesvirus 3 group, human herpesvirus 4
group, human herpesvirus 6, human herpesvirus 8, human
immunodeficiency virus, human immunodeficiency virus 1, human
immunodeficiency virus 2, human papillomavirus, human T cell
leukemia virus, human T cell leukemia virus I, human T cell
leukemia virus II, human T cell leukemia virus III, human T cell
lymphoma virus I, human T cell lymphoma virus II, human T cell
lymphotropic virus type 1, human T cell lymphotropic virus type 2,
human T lymphotropic virus I, human T lymphotropic virus II, human
T lymphotropic virus III, Ichnovirus, infantile gastroenteritis
virus, infectious bovine rhinotracheitis virus, infectious
haematopoietic necrosis virus, infectious pancreatic necrosis
virus, influenza virus A, influenza virus B, influenza virus C,
influenza virus D, influenza virus pr8, insect iridescent virus,
insect virus, iridovirus, Japanese B virus, Japanese encephalitis
virus, JC virus, Junin virus, Kaposi's sarcoma-associated
herpesvirus, Kemerovo virus, Kilham's rat virus, Klamath virus,
Kolongo virus, Korean hemorrhagic fever virus, kumba virus, Kysanur
forest condition virus, Kyzylagach virus, La Crosse virus, lactic
dehydrogenase elevating virus, lactic dehydrogenase virus, Lagos
bat virus, Langur virus, lapine parvovirus, Lassa fever virus,
Lassa virus, latent rat virus, LCM virus, Leaky virus, Lentivirus,
Leporipoxvirus, leukemia virus, leukovirus, lumpy skin condition
virus, lymphadenopathy associated virus, Lymphocryptovirus,
lymphocytic choriomeningitis virus, lymphoproliferative virus
group, Machupo virus, mad itch virus, mammalian type B oncovirus
group, mammalian type B retroviruses, mammalian type C retrovirus
group, mammalian type D retroviruses, mammary tumor virus, Mapuera
virus, Marburg virus, Marburg-like virus, Mason Pfizer monkey
virus, Mastadenovirus, Mayaro virus, ME virus, measles virus,
Menangle virus, Mengo virus, Mengovirus, Middelburg virus, milkers
nodule virus, mink enteritis virus, minute virus of mice, MLV
related virus, MM virus, Mokola virus, Molluscipoxvirus, Molluscum
contagiosum virus, monkey B virus, monkeypox virus,
Mononegavirales, Morbillivirus, Mount Elgon bat virus, mouse
cytomegalovirus, mouse encephalomyelitis virus, mouse hepatitis
virus, mouse K virus, mouse leukemia virus, mouse mammary tumor
virus, mouse minute virus, mouse pneumonia virus, mouse
poliomyelitis virus, mouse polyomavirus, mouse sarcoma virus,
mousepox virus, Mozambique virus, Mucambo virus, mucosal condition
virus, mumps virus, murid betaherpesvirus 1, murid cytomegalovirus
2, murine cytomegalovirus group, murine encephalomyelitis virus,
murine hepatitis virus, murine leukemia virus, murine nodule
inducing virus, murine polyomavirus, murine sarcoma virus,
Muromegalovirus, Murray Valley encephalitis virus, myxoma virus,
Myxovirus, Myxovirus multiforme, Myxovirus parotitidis, Nairobi
sheep condition virus, Nairovirus, Nanirnavirus, Nariva virus,
Ndumo virus, Neethling virus, Nelson Bay virus, neurotropic virus,
New World Arenavirus, newborn pneumonitis virus, Newcastle
condition virus, Nipah virus, noncytopathogenic virus, Norwalk
virus, nuclear polyhedrosis virus (NPV), nipple neck virus,
O'nyong'nyong virus, Ockelbo virus, oncogenic virus, oncogenic
viruslike particle, oncornavirus, Orbivirus, Orf virus, Oropouche
virus, Orthohepadnavirus, Orthomyxovirus, Orthopoxvirus,
Orthoreovirus, Orungo, ovine papillomavirus, ovine catarrhal fever
virus, owl monkey herpesvirus, Palyam virus, Papillomavirus,
Papillomavirus sylvilagi, Papovavirus, parainfluenza virus,
parainfluenza virus type 1, parainfluenza virus type 2,
parainfluenza virus type 3, parainfluenza virus type 4,
Paramyxovirus, Parapoxvirus, paravaccinia virus, Parvovirus,
Parvovirus B19, parvovirus group, Pestivirus, Phlebovirus, phocine
distemper virus, Picodnavirus, Picornavirus, pig
cytomegalovirus-pigeonpox virus, Piry virus, Pixuna virus,
pneumonia virus of mice, Pneumovirus, poliomyelitis virus,
poliovirus, Polydnavirus, polyhedral virus, polyoma virus,
Polyomavirus, Polyomavirus bovis, Polyomavirus cercopitheci,
Polyomavirus hominis 2, Polyomavirus maccacae 1, Polyomavirus muris
1, Polyomavirus muris 2, Polyomavirus papionis 1, Polyomavirus
papionis 2, Polyomavirus sylvilagi, Pongine herpesvirus 1, porcine
epidemic diarrhea virus, porcine hemagglutinating encephalomyelitis
virus, porcine parvovirus, porcine transmissible gastroenteritis
virus, porcine type C virus, pox virus, poxvirus, poxvirus
variolae, Prospect Hill virus, Provirus, pseudocowpox virus,
pseudorabies virus, psittacinepox virus, quailpox virus, rabbit
fibroma virus, rabbit kidney vaculolating virus, rabbit
papillomavirus, rabies virus, raccoon parvovirus, raccoonpox virus,
Ranikhet virus, rat cytomegalovirus, rat parvovirus, rat virus,
Rauscher's virus, recombinant vaccinia virus, recombinant virus,
reovirus, reovirus 1, reovirus 2, reovirus 3, reptilian type C
virus, respiratory infection virus, respiratory syncytial virus,
respiratory virus, reticuloendotheliosis virus, Rhabdovirus,
Rhabdovirus carpia, Rhadinovirus, Rhinovirus, Rhizidiovirus, Rift
Valley fever virus, Riley's virus, rinderpest virus, RNA tumor
virus, Ross River virus, Rotavirus, rougeole virus, Rous sarcoma
virus, rubella virus, rubeola virus, Rubivirus, Russian autumn
encephalitis virus, SA 11 simian virus, SA2 virus, Sabia virus,
Sagiyama virus, Saimirine herpesvirus 1, salivary gland virus,
sandfly fever virus group, Sandjimba virus, SARS virus, SDAV
(sialodacryoadenitis virus), sealpox virus, Semliki Forest Virus,
Seoul virus, sheeppox virus, Shope fibroma virus, Shope papilloma
virus, simian foamy virus, simian hepatitis A virus, simian human
immunodeficiency virus, simian immunodeficiency virus, simian
parainfluenza virus, simian T cell lymphotrophic virus, simian
virus, simian virus 40, Simplexvirus, Sin Nombre virus, Sindbis
virus, smallpox virus, South American hemorrhagic fever viruses,
sparrowpox virus, Spumavirus, squirrel fibroma virus, squirrel
monkey retrovirus, SSV 1 virus group, STLV (simian T lymphotropic
virus) type I, STLV (simian T lymphotropic virus) type II, STLV
(simian T lymphotropic virus) type III, stomatitis papulosa virus,
submaxillary virus, suid alphaherpesvirus 1, suid herpesvirus 2,
Suipoxvirus, swamp fever virus, swinepox virus, Swiss mouse
leukemia virus, TAC virus, Tacaribe complex virus, Tacaribe virus,
Tanapox virus, Taterapox virus, Tench reovirus, Theiler's
encephalomyelitis virus, Theiler's virus, Thogoto virus,
Thottapalayam virus, Tick borne encephalitis virus, Tioman virus,
Togavirus, Torovirus, tumor virus, Tupaia virus, turkey
rhinotracheitis virus, turkeypox virus, type C retroviruses, type D
oncovirus, type D retrovirus group, ulcerative condition
rhabdovirus, Una virus, Uukuniemi virus group, vaccinia virus,
vacuolating virus, varicella zoster virus, Varicellovirus, Varicola
virus, variola major virus, variola virus, Vasin Gishu condition
virus, VEE virus, Venezuelan equine encephalitis virus, Venezuelan
equine encephalomyelitis virus, Venezuelan hemorrhagic fever virus,
vesicular stomatitis virus, Vesiculovirus, Vilyuisk virus, viper
retrovirus, viral haemorrhagic septicemia virus, Visna Maedi virus,
Visna virus, volepox virus, VSV (vesicular stomatitis virus),
Wallal virus, Warrego virus, wart virus, WEE virus, West Nile
virus, western equine encephalitis virus, western equine
encephalomyelitis virus, Whataroa virus, Winter Vomiting Virus,
woodchuck hepatitis B virus, woolly monkey sarcoma virus, wound
tumor virus, WRSV virus, Yaba monkey tumor virus, Yaba virus,
Yatapoxvirus, yellow fever virus, and the Yug Bogdanovac virus.
Inflammatory Conditions
[0156] Inflammatory conditions that can be treated and/or prevented
using the methods and compositions provided herein include, for
example, autoimmune condition. Autoimmune conditions include, for
example, rheumatoid arthritis, multiple sclerosis, Sjogren's
syndrome, systemic lupus erythematosus, autoimmune hepatitis,
autoimmune thyroiditis, hemophagocytic syndrome (hemophagocytic
lymphohistiocytosis), diabetes mellitus type 1, Crohn's condition,
ulcerative colitis, psoriasis, psoriatic arthritis, idiopathic
thrombocytonpenic pupura, polymyositis, dermatomyositis, myasthenia
gravis, autoimmune thryroiditis, Evan's syndrome, autoimmune
hemolytic anemia, aplastic anemia, autoimmune neutropenia,
scleroderma, Reiter's syndrome, ankylosing spondylitis, pemphnigus,
pemphigoid or autoimmune hepatitis, Behcet's condition, Celiac
condition, Chagas condition, acute disseminated encephalomyelitis,
Addison's condition, antiphospholipid antibody syndrome, autoimmune
inner ear condition, bullous pemphigoid, Chronic obstructive
pulmonary condition, Goodpasture's syndrome, Graves' condition,
Guillain-Barre syndrome, Hashimoto's thyroditis, Hidradenitis
suppurativa, Interstitial cystitis, neuromyotonia, pemphigus
vulgaris, pernicious anemia, primary biliary cirrhosis, and
vasculitis syndromes.
[0157] Other inflammatory conditions that can be treated and/or
prevented using the methods and compositions of the provided
invention include, for example, an allergic condition (e.g.,
allergic rhinitis, asthma, atopic eczema), a skin condition,
coronary artery condition, peripheral artery condition,
atherosclerosis, retinitis, pancreatitis, cardiomyopathy,
pericarditis, colitis, glomerulonephritis, lung inflammation,
esophagitis, gastritis, duodenitis, ileitis, meningitis,
encephalitis, encephalomyelitis, transverse myelitis, cystitis,
urethritis, mucositis, lymphadenitis, dermatitis, hepatitis,
osteomyelitis, or herpes zoster.
Formulations, Routes of Administration, and Effective Doses
[0158] Another aspect of the present invention relates to
formulations, routes of administration and effective doses for
pharmaceutical compositions comprising an agent or combination of
agents. Such pharmaceutical compositions can be used to treat a
virus-induced inflammatory condition as described above. A
pharmaceutical composition can comprise a viral inducing agent. A
pharmaceutical composition can comprise a viral inducing agent and
one or more additional agents. A pharmaceutical composition can
comprise an antiviral agent. A pharmaceutical composition can
comprise an antiviral agent and one or more additional agents. A
pharmaceutical composition can comprise a viral inducing agent and
an antiviral agent. A pharmaceutical composition can comprise a
viral inducing agent, an antiviral agent, and one or more
additional agents.
[0159] The agents or their pharmaceutically acceptable salts can be
provided alone or in combination with one or more other agents or
with one or more other forms. For example, a formulation can
comprise one or more agents in particular proportions, depending on
the relative potencies of each agent and the intended indication.
For example, in compositions for targeting two different targets
and where potencies are similar, about a 1:1 ratio of agents can be
used. The two forms can be formulated together, in the same dosage
unit e.g. in one cream, suppository, tablet, capsule, enteric
coated tablet or capsule, aerosol spray, or packet of powder to be
dissolved in a beverage; or each form may be formulated in a
separate unit, e.g., two creams, two suppositories, two tablets,
two capsules, a tablet and a liquid for dissolving the tablet, two
aerosol sprays, or a packet of powder and a liquid for dissolving
the powder, etc.
[0160] A "pharmaceutically acceptable salt" can be a salt that
retains the biological effectiveness and properties of one or more
agents, and which are not biologically or otherwise undesirable.
For example, a pharmaceutically acceptable salt does not interfere
with the beneficial effect of a viral inducing agent or an
antiviral agent.
[0161] Salts can include those of the inorganic ions, for example,
sodium, potassium, calcium, magnesium ions, and the like. Salts can
include salts with inorganic or organic acids, for example,
hydrochloric acid, hydrobromic acid, phosphoric acid, nitric acid,
sulfuric acid, methanesulfonic acid, p-toluenesulfonic acid, acetic
acid, fumaric acid, succinic acid, lactic acid, mandelic acid,
malic acid, citric acid, tartaric acid or maleic acid. If one or
more agents contain a carboxy group or other acidic group, it can
be converted into a pharmaceutically acceptable addition salt with
inorganic or organic bases. Examples of suitable bases include
sodium hydroxide, potassium hydroxide, ammonia, cyclohexylamine,
dicyclohexyl-amine, ethanolamine, diethanolamine, triethanolamine,
and the like.
[0162] A pharmaceutically acceptable ester or amide can be an ester
or amide that retains biological effectiveness and properties of
one or more agents, and which are not biologically or otherwise
undesirable. For example, the ester or amide does not interfere
with the beneficial effect of a viral inducing agent, an antiviral
agent, or an additional agent. Esters can include, for example,
ethyl, methyl, isobutyl, ethylene glycol, and the like. Amides
include can include, for example, unsubstituted amides, alkyl
amides, dialkyl amides, and the like.
[0163] A viral inducing agent, for example a HDAC inhibitor, can be
administered in combination with an antiviral agent. Pharmaceutical
compositions comprising a combination of a viral inducing agent and
an antiviral agent can be formulated to comprise certain molar
ratios. For example, molar ratios of about 99:1 to about 1:99 of a
viral inducing agent to the antiviral agent can be used. The range
of molar ratios of viral inducing agent: the antiviral agent can be
selected from about 80:20 to about 20:80; about 75:25 to about
25:75, about 70:30 to about 30:70, about 66:33 to about 33:66,
about 60:40 to about 40:60; about 50:50; and about 90:10 to about
10:90. The viral inducing agent and the antiviral agent can be
co-formulated, in the same dosage unit, e.g., in one cream,
suppository, tablet, capsule, enteric coated capsule or tablet, or
packet of powder to be dissolved in a beverage; or each agent,
form, and/or compound can be formulated in separate units, e.g.,
two creams, suppositories, tablets, two capsules, enteric coated
capsules or tablets, a tablet and a liquid for dissolving the
tablet, an aerosol spray a packet of powder and a liquid for
dissolving the powder, etc.
[0164] An agent can be administered in combination with one or more
other compounds, forms, and/or agents, e.g., as described above.
Pharmaceutical compositions comprising combinations of a viral
inducing agent and/or antiviral agent with one or more other active
agents can be formulated to comprise certain molar ratios. For
example, molar ratios of about 99:1 to about 1:99 of a viral
inducing agent to the other active agent can be used; molar ratios
of about 99:1 to about 1:99 of an antiviral agent to the other
active agent can be used; molar ratios of about 99:1 to about 1:99
of a viral inducing agent and antiviral agent can be used. The
range of molar ratios of viral inducing agent: other active agent
can be selected from about 80:20 to about 20:80; about 75:25 to
about 25:75, about 70:30 to about 30:70, about 66:33 to about
33:66, about 60:40 to about 40:60; about 50:50; and about 90:10 to
about 10:90. The range of molar ratios of an antiviral agent: other
active agent can be selected from about 80:20 to about 20:80; about
75:25 to about 25:75, about 70:30 to about 30:70, about 66:33 to
about 33:66, about 60:40 to about 40:60; about 50:50; and about
90:10 to about 10:90. The molar ratio may of a viral inducing
agent:other active agent can be about 1:9 or about 1:1. The molar
ratio may of an antiviral agent:other active agent can be about 1:9
or about 1:1. Two or more agents, forms and/or compounds can be
formulated together, in the same dosage unit, e.g., in one cream,
suppository, tablet, capsule, enteric coated capsule or tablet, or
packet of powder to be dissolved in a beverage; or each agent,
form, and/or compound can be formulated in separate units, e.g.,
two creams, suppositories, tablets, two capsules, enteric coated
capsules or tablets, a tablet and a liquid for dissolving the
tablet, an aerosol spray a packet of powder and a liquid for
dissolving the powder, etc.
[0165] A viral inducing agent, for example a HDAC inhibitor, can be
administered in combination with an antiviral agent. Pharmaceutical
compositions comprising a combination of a viral inducing agent and
an antiviral agent can be formulated to comprise certain mg per
dose. For example, a viral inducing agent can be administered at
0.01, 0.05, 0.1, 0.5, 1, 2, 5, 10, 20, 25, 50, 100, 250, 500, 1000
mg/kg per dose. A HDAC inhibitor can be administered at 0.01-0.1,
0.05-0.5, 1-2, 1-5, 5-10, 10-20, 10-25, 10-50, 100-500, or 500-1000
mg/kg per dose. A single dose of an oral formulation of a viral
inducing agent can contain 0.01, 0.05, 0.1, 0.5, 1, 2, 5, 10, 20,
25, 50, 100, 250, 500, 1000 mg. In one embodiment, the HDAC
inhibitor is administered at 0.01, 0.05, 0.1, 0.5, 1, 2, 5, 10, 20,
25, 50, 100, 250, 500, 1000 mg/kg per dose. In a related
embodiment, the HDAC inhibitor is administered orally. In a
specific embodiment, the total daily oral dose of a HDAC inhibitor
is no more than 1, 2, 5, 10, 20, 25, 40, 50, 100, 250, or 500 mg.
In another related embodiment, the HDAC inhibitor is administered
1, 2, 3, 4, or 5 times a day orally. In other embodiments, a single
daily dose of a HDAC inhibitor is provided whereas oral
valganciclovir is provided at 900 mgs/dose, two times a day.
[0166] An oral formulation of an HDAC inhibitor can be
co-formulated with an antiviral agent, such as valganciclovir. In a
specific embodiment when an HDAC inhibitor and valganciclovir are
co-formulated for a single daily dose, the valganciclovir is
present in a slow release or timed release form. In certain
embodiments, the HDAC inhibitor and valganciclovir or other
antiviral agent are co-formulated such that the HDAC inhibitor is
present at no more than 100 mg per dose, and the antiviral agent is
present at no more than 1000 mg per dose. In some embodiments, the
HDAC inhibitor and valganciclovir or other antiviral agent are
co-formulated such that the HDAC inhibitor is present at no more
than 80 mg per dose, and the antiviral agent is present at no more
than 500 mg per dose. In certain embodiments, the HDAC inhibitor
and valganciclovir or other antiviral agent are co-formulated such
that the HDAC inhibitor is present at not greater than 80 mg per
dose, and the antiviral agent is present at not greater than 1500
mg per dose.
[0167] In some embodiments, a co-formulation comprising an HDAC
inhibitor and an antiviral agent comprises less than 500 mg, less
than 400 mg, less than 300 mg, less than 200 mg, less than 100 mg,
less than 90 mg, less than 80 mg, less than 70 mg, less than 60 mg,
less than 50 mg, less than 40 mg, less than 30 mg, less than 20 mg,
less than 10 mg, less than 5 mg, less than 2 mg, or less than 1 mg
of the HDAC inhibitor. In other embodiments, a co-formulation
comprising an HDAC inhibitor and an antiviral agent comprises less
than 1500 mg, less than 1400 mg, less than 1300 mg, less than 1200
mg, less than 1100 mg, less than 1000 mg, less than 900 mg, less
than 800 mg, less than 700 mg, less than 600 mg, less than 500 mg,
less than 400 mg, less than 300 mg, less than 200 mg, less than 100
mg, less than 90 mg, less than 80 mg, less than 70 mg, less than 60
mg, less than 50 mg, less than 40 mg, less than 30 mg, less than 20
mg, less than 10 mg, less than 5 mg, less than 2 mg, or less than 1
mg of the antiviral agent.
[0168] In certain embodiments, a unit dose of a co-formulated HDAC
inhibitor and antiviral agent comprises between about 1 mg and
about 500 mg of the HDAC inhibitor and between 1 mg and 1500 mg of
the antiviral agent. In some embodiments, the unit dose comprises
about 500 mg, about 400 mg, about 300 mg, about 200 mg, about 100
mg, about 90 mg, about 80 mg, about 70 mg, about 60 mg, about 50
mg, about 40 mg, about 30 mg, about 20 mg, about 10 mg, about 5 mg,
about 2 mg, or about 1 mg of the HDAC inhibitor. In certain
embodiments, the unit dose comprises less than 500 mg, less than
400 mg, less than 300 mg, less than 200 mg, less than 100 mg, less
than 90 mg, less than 80 mg, less than 70 mg, less than 60 mg, less
than 50 mg, less than 40 mg, less than 30 mg, less than 20 mg, less
than 10 mg, less than 5 mg, less than 2 mg, or less than 1 mg of an
HDAC inhibitor. In some embodiments, the unit dose comprises more
than 500 mg, more than 400 mg, more than 300 mg, more than 200 mg,
more than 100 mg, more than 90 mg, more than 80 mg, more than 70
mg, more than 60 mg, more than 50 mg, more than 40 mg, more than 30
mg, more than 20 mg, more than 10 mg, more than 5 mg, more than 2
mg, or more than 1 mg of an HDAC inhibitor. In certain embodiments,
the unit dose comprises more than 2 mg and less than 500 mg of an
HDAC inhibitor. In some embodiments, the unit dose comprises more
than 10 mg and less than 50 mg of an HDAC inhibitor.
[0169] In some embodiments, the unit dose comprises about 2000 mg,
about 1900 mg, about 1800 mg, about 1700 mg, about 1600 mg,
comprises about 1500 mg, about 1400 mg, about 1300 mg, about 1200
mg, about 1100 mg, about 1000 mg, about 900 mg, about 800 mg, about
750 mg, about 700 mg, about 650 mg, about 600 mg, about 550 mg,
about 500 mg, about 450 mg, about 400 mg, about 350 mg, about 300
mg, about 250 mg, about 200 mg, about 150 mg, about 140 mg, about
130 mg, about 120 mg, about 110 mg, about 100 mg, about 90 mg,
about 80 mg, about 70 mg, about 60 mg, about 50 mg, about 40 mg,
about 30 mg, about 20 mg, about 10 mg, about 5 mg, about 2 mg, or
about 1 mg of the antiviral agent. In certain embodiments, the unit
dose comprises less than 2000 mg, less than 1900 mg, less than 1800
mg, less than 1700 mg, less than 1600 mg, comprises less than 1500
mg, less than 1400 mg, less than 1300 mg, less than 1200 mg, less
than 1100 mg, less than 1000 mg, less than 900 mg, less than 800
mg, less than 750 mg, less than 700 mg, less than 650 mg, less than
600 mg, less than 550 mg, less than 500 mg, less than 450 mg, less
than 400 mg, less than 350 mg, less than 300 mg, less than 250 mg,
less than 200 mg, less than 150 mg, less than 140 mg, less than 130
mg, less than 120 mg, less than 110 mg, less than 100 mg, less than
90 mg, less than 80 mg, less than 70 mg, less than 60 mg, less than
50 mg, less than 40 mg, less than 30 mg, less than 20 mg, less than
10 mg, less than 5 mg, less than 2 mg, or less than 1 mg of the
antiviral agent. In some embodiments, the unit dose comprises more
than 2000 mg, more than 1900 mg, more than 1800 mg, more than 1700
mg, more than 1600 mg, comprises more than 1500 mg, more than 1400
mg, more than 1300 mg, more than 1200 mg, more than 1100 mg, more
than 1000 mg, more than 900 mg, more than 800 mg, more than 750 mg,
more than 700 mg, more than 650 mg, more than 600 mg, more than 550
mg, more than 500 mg, more than 450 mg, more than 400 mg, more than
350 mg, more than 300 mg, more than 250 mg, more than 200 mg, more
than 150 mg, more than 140 mg, more than 130 mg, more than 120 mg,
more than 110 mg, more than 100 mg, more than 90 mg, more than 80
mg, more than 70 mg, more than 60 mg, more than 50 mg, more than 40
mg, more than 30 mg, more than 20 mg, more than 10 mg, more than 5
mg, more than 2 mg, or more than 1 mg of the antiviral agent. In
certain embodiments, the unit dose comprises more than 50 mg and
less than 1500 mg of the antiviral agent. In some embodiments, the
unit dose comprises more than 100 mg and less than 500 mg of the
antiviral agent. In certain embodiments, the antiviral agent is
formulated as slow release.
[0170] In some embodiments, the co-formulated HDAC inhibitor and
antiviral agent are administered once a day. In certain
embodiments, the co-formulated HDAC inhibitor and antiviral agent
are administered twice a day. In other embodiments, the
co-formulated HDAC inhibitor and antiviral agent are administered
thrice a day. In some embodiments, the co-formulated HDAC inhibitor
and antiviral agent are administered once a day, twice a day, or
thrice a day, and a further dose of the HDAC inhibitor is
administered once, twice, or thrice a day. In certain embodiments,
the co-formulated HDAC inhibitor and antiviral agent are
administered once a day, twice a day, or thrice a day, and a
further dose of the antiviral agent is administered once, twice, or
thrice a day.
[0171] In certain embodiments, one unit dose of the co-formulated
HDAC inhibitor and antiviral agent are administered per day. In
some embodiments, two unit doses of the co-formulated HDAC
inhibitor and antiviral agent are administered per day. In certain
embodiments, three unit doses of the co-formulated HDAC inhibitor
and antiviral agent are administered per day. In some embodiments,
four unit doses of the co-formulated HDAC inhibitor and antiviral
agent are administered per day. In certain embodiments, the one,
two, three, or four unit doses are administered daily, once a week,
twice a week, three times a week, four times a week, or five times
a week.
[0172] In some embodiments, one or more unit doses of the
co-formulated HDAC inhibitor and antiviral agent are administered
in combination with other treatments, such as antibodies,
chemotherapy drugs, and radiation therapy.
[0173] One or more agents and/or combinations of agents can be
administered with still other agents. The choice of agents that can
be co-administered with the agents and/or combinations of agents
can depend, at least in part, on the condition being treated.
Agents of particular use in the formulations of the present
invention include, for example, any agent having a therapeutic
effect for a virus-induced inflammatory condition, including, e.g.,
drugs used to treat inflammatory conditions. For example,
formulations of the instant invention can additionally contain one
or more conventional anti-inflammatory drugs, such as an NSAID,
e.g. ibuprofen, naproxen, acetominophen, ketoprofen, or aspirin. In
some alternative embodiments for the treatment of a virus-induced
inflammatory condition can additionally contain one or more
conventional influenza antiviral agents, such as amantadine,
rimantadine, zanamivir, and oseltamivir. In treatments for
retroviral infections, such as HIV, formulations of the instant
invention may additionally contain one or more conventional
antiviral drug, such as protease inhibitors (lopinavir/ritonavir
{Kaletra.TM.}, indinavir {Crixivan.TM.}, ritonavir {Norvir.TM.},
nelfinavir {Viracept.TM.}, saquinavir hard gel capsules
{Invirase.TM.}, atazanavir {Reyataz.TM.}, amprenavir
{Agenerase.TM.}, fosamprenavir {Telzir.TM.}, tipranavir
{Aptivus.TM. }), reverse transcriptase inhibitors,
includingnon-Nucleoside and Nucleoside/nucleotide inhibitors (AZT
{zidovudine, Retrovir.TM.}, ddI {didanosine, Videx.TM.}, 3TC
{lamivudine, Epivir.TM.}, d4T {stavudine, Zerit.TM.}, abacavir
{Ziagen.TM.}, FTC {emtricitabine, Emtriva.TM.}, tenofovir
{Viread.TM.}, efavirenz {Sustiva.TM.} and nevirapine
{Viramune.TM.}), fusion inhibitors T20 {enfuvirtide, Fuzeon.TM.}.,
integrase inhibitors (MK-0518 and GS-9137), and maturation
inhibitors (PA-457 {Bevirimat.TM.}). As another example,
formulations can additionally contain one or more supplements, such
as vitamin C, E or other anti-oxidants.
[0174] One or more agents (or pharmaceutically acceptable salts,
esters or amides thereof) can be administered per se or in the form
of a pharmaceutical composition wherein the one or more active
agent(s) is in an admixture or mixture with one or more
pharmaceutically acceptable carriers. A pharmaceutical composition,
as used herein, can be any composition prepared for administration
to a subject. Pharmaceutical compositions can be formulated in
conventional manner using one or more physiologically acceptable
carriers, comprising excipients, diluents, and/or auxiliaries,
e.g., that facilitate processing of the active agents into
preparations that can be administered. Proper formulation can
depend at least in part upon the route of administration chosen.
One or more agents, or pharmaceutically acceptable salts, esters,
or amides thereof, can be delivered to a patient using a number of
routes or modes of administration, including oral, buccal, topical,
rectal, transdermal, transmucosal, subcutaneous, intravenous, and
intramuscular applications, as well as by inhalation.
[0175] For oral administration, one or more agents can be
formulated readily by combining the one or more active agents with
pharmaceutically acceptable carriers well known in the art. Such
carriers can enable the one or more agents to be formulated as
tablets, including chewable tablets, pills, dragees, capsules,
lozenges, hard candy, liquids, gels, syrups, slurries, powders,
suspensions, elixirs, wafers, and the like, for oral ingestion by a
patient to be treated. Such formulations can comprise
pharmaceutically acceptable carriers including solid diluents or
fillers, sterile aqueous media and various non-toxic organic
solvents. Generally, the agents of the invention can be included at
concentration levels ranging from about 0.5%, about 5%, about 10%,
about 20%, or about 30% to about 50%, about 60%, about 70%, about
80% or about 90% by weight of the total composition of oral dosage
forms, in an amount sufficient to provide a desired unit of
dosage.
[0176] Aqueous suspensions for oral use can contain one or more
agents with pharmaceutically acceptable excipients, such as a
suspending agent (e.g., methyl cellulose), a wetting agent (e.g.,
lecithin, lysolecithin and/or a long-chain fatty alcohol), as well
as coloring agents, preservatives, flavoring agents, and the
like.
[0177] Oils or non-aqueous solvents can be required to bring one or
more agents into solution, due to, for example, the presence of
large lipophilic moieties. Alternatively, emulsions, suspensions,
or other preparations, for example, liposomal preparations, can be
used. With respect to liposomal preparations, any known methods for
preparing liposomes for treatment of a condition can be used. See,
for example, Bangham et al., J. Mol. Biol. 23: 238-252 (1965) and
Szoka et al., Proc. Natl Acad. Sci. USA 75: 4194-4198 (1978),
incorporated herein by reference. Ligands can also be attached to
the liposomes to direct these compositions to particular sites of
action. One or more agents can also be integrated into foodstuffs,
e.g, cream cheese, butter, salad dressing, or ice cream to
facilitate solubilization, administration, and/or compliance in
certain patient populations.
[0178] Pharmaceutical preparations for oral use can be obtained as
a solid excipient, optionally grinding a resulting mixture, and
processing the mixture of granules, after adding suitable
auxiliaries, if desired, to obtain tablets or dragee cores.
Suitable excipients are, in particular, fillers such as sugars,
including lactose, sucrose, mannitol, or sorbitol; flavoring
elements, cellulose preparations such as, for example, maize
starch, wheat starch, rice starch, potato starch, gelatin, gum
tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium
carboxymethylcellulose, and/or polyvinyl pyrrolidone (PVP).
Disintegrating agents can be added, for example, the cross-linked
polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof such
as sodium alginate. One or more agents can also be formulated as a
sustained release preparation.
[0179] Dragee cores can be provided with suitable coatings. For
this purpose, concentrated sugar solutions may be used, which may
optionally contain gum arabic, talc, polyvinyl pyrrolidone,
carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquer
solutions, and suitable organic solvents or solvent mixtures.
Dyestuffs or pigments can be added to the tablets or dragee
coatings for identification or to characterize different
combinations of one or more active agents.
[0180] Pharmaceutical preparations that can be used orally include
push-fit capsules made of gelatin, as well as soft, sealed capsules
made of gelatin and a plasticizer, such as glycerol or sorbitol.
The push-fit capsules can contain the active ingredients in
admixture with filler such as lactose, binders such as starches,
and/or lubricants such as talc or magnesium stearate and,
optionally, stabilizers. In soft capsules, the active agents can be
dissolved or suspended in suitable liquids, such as fatty oils,
liquid paraffin, or liquid polyethylene glycols. In addition,
stabilizers can be added. All formulations for oral administration
can be in dosages suitable for administration.
[0181] For injection, one or more agents can be formulated in
aqueous solutions, including but not limited to physiologically
compatible buffers such as Hank's solution, Ringer's solution, or
physiological saline buffer. Such compositions can also include one
or more excipients, for example, preservatives, solubilizers,
fillers, lubricants, stabilizers, albumin, and the like. Methods of
formulation are known in the art, for example, as disclosed in
Remington's Pharmaceutical Sciences, latest edition, Mack
Publishing Co., Easton P.
[0182] One or more agents can also be formulated as a depot
preparation. Such long acting formulations can be administered by
implantation or transcutaneous delivery (for example subcutaneously
or intramuscularly), intramuscular injection or use of a
transdermal patch. Thus, for example, one or more agents can be
formulated with suitable polymeric or hydrophobic materials (for
example as an emulsion in an acceptable oil) or ion exchange
resins, or as sparingly soluble derivatives, for example, as a
sparingly soluble salt.
[0183] Pharmaceutical compositions comprising one or more agents
can exert local and regional effects when administered topically or
injected at or near particular sites of infection. Direct topical
application, e.g., of a viscous liquid, gel, jelly, cream, lotion,
ointment, suppository, foam, or aerosol spray, can be used for
local administration, to produce, for example local and/or regional
effects. Pharmaceutically appropriate vehicles for such formulation
include, for example, lower aliphatic alcohols, polyglycols (e.g.,
glycerol or polyethylene glycol), esters of fatty acids, oils,
fats, silicones, and the like. Such preparations may also include
preservatives (e.g., p-hydroxybenzoic acid esters) and/or
antioxidants (e.g., ascorbic acid and tocopherol). See also
Dermatological Formulations: Percutaneous absorption, Barry (Ed.),
Marcel Dekker Incl, 1983. In some embodiments, local/topical
formulations comprising a viral inducing agent and or antiviral
agent are used to treat epidermal or mucosal viral-induced
inflammatory condition.
[0184] Pharmaceutical compositions can contain a cosmetically or
dermatologically acceptable carrier. Such carriers can be
compatible with skin, nails, mucous membranes, tissues and/or hair,
and can include any conventionally used cosmetic or dermatological
carrier meeting these requirements. Such carriers can be readily
selected by one of ordinary skill in the art. In formulating skin
ointments, an agent or combination of agents can be formulated in
an oleaginous hydrocarbon base, an anhydrous absorption base, a
water-in-oil absorption base, an oil-in-water water-removable base
and/or a water-soluble base.
[0185] The compositions according to the present invention can be
in any form suitable for topical application, including aqueous,
aqueous-alcoholic or oily solutions, lotion or serum dispersions,
aqueous, anhydrous or oily gels, emulsions obtained by dispersion
of a fatty phase in an aqueous phase (O/W or oil in water) or,
conversely, (W/O or water in oil), microemulsions or alternatively
microcapsules, microparticles or lipid vesicle dispersions of ionic
and/or nonionic type. These compositions can be prepared according
to conventional methods. The amounts of the various constituents of
the compositions according to the invention can be those
conventionally used in the art. These compositions constitute
protection, treatment or care creams, milks, lotions, gels or foams
for the face, for the hands, for the body and/or for the mucous
membranes, or for cleansing the skin. The compositions can also
consist of solid preparations constituting soaps or cleansing
bars.
[0186] A pharmaceutical composition can also contain adjuvants
common to the cosmetic and dermatological fields, for example,
hydrophilic or lipophilic gelling agents, hydrophilic or lipophilic
active agents, preserving agents, antioxidants, solvents,
fragrances, fillers, sunscreens, odor-absorbers and dyestuffs. The
amounts of these various adjuvants can be those conventionally used
in the fields considered and, for example, are from about 0.01% to
about 20% of the total weight of the composition. Depending on
their nature, these adjuvants can be introduced into the fatty
phase, into the aqueous phase and/or into the lipid vesicles.
[0187] Ocular viral infections can be effectively treated with
ophthalmic solutions, suspensions, ointments or inserts comprising
an agent or combination of agents of the present invention.
[0188] In some embodiments, viral infections of the ear can be
effectively treated with otic solutions, suspensions, ointments or
inserts comprising an agent or combination of agents of the present
invention.
[0189] One or more agents can be delivered in soluble rather than
suspension form, which can allow for more rapid and quantitative
absorption to the sites of action. In general, formulations such as
jellies, creams, lotions, suppositories and ointments can provide
an area with more extended exposure to the agents of the present
invention, while formulations in solution, e.g., sprays, provide
more immediate, short-term exposure.
[0190] Relating to topical/local application, a pharmaceutical
composition can include one or more penetration enhancers. For
example, the formulations can comprise suitable solid or gel phase
carriers or excipients that increase penetration or help delivery
of agents or combinations of agents of the invention across a
permeability barrier, e.g., the skin. Many of these
penetration-enhancing compounds are known in the art of topical
formulation, and include, e.g., water, alcohols (e.g., terpenes
like methanol, ethanol, 2-propanol), sulfoxides (e.g., dimethyl
sulfoxide, decylmethyl sulfoxide, tetradecylmethyl sulfoxide),
pyrrolidones (e.g., 2-pyrrolidone, N-methyl-2-pyrrolidone,
N-(2-hydroxyethyl)pyrrolidone), laurocapram, acetone,
dimethylacetamide, dimethylformamide, tetrahydrofurfuryl alcohol,
L-.alpha.-amino acids, anionic, cationic, amphoteric or nonionic
surfactants (e.g., isopropyl myristate and sodium lauryl sulfate),
fatty acids, fatty alcohols (e.g., oleic acid), amines, amides,
clofibric acid amides, hexamethylene lauramide, proteolytic
enzymes, .alpha.-bisabolol, d-limonene, urea and
N,N-diethyl-m-toluamide, and the like. Additional examples include
humectants (e.g., urea), glycols (e.g., propylene glycol and
polyethylene glycol), glycerol monolaurate, alkanes, alkanols,
ORGELASE, calcium carbonate, calcium phosphate, various sugars,
starches, cellulose derivatives, gelatin, and/or other polymers. A
pharmaceutical composition can include one or more such penetration
enhancers.
[0191] A pharmaceutical composition for local/topical application
can include one or more antimicrobial preservatives, for example,
quaternary ammonium compounds, organic mercurials, p-hydroxy
benzoates, aromatic alcohols, chlorobutanol, and the like.
[0192] Gastrointestinal viral infections can be effectively treated
with orally- or rectally delivered solutions, suspensions,
ointments, enemas and/or suppositories comprising an agent or
combination of agents of the present invention.
[0193] Respiratory viral infections can be effectively treated with
aerosol solutions, suspensions or dry powders comprising an agent
or combination of agents of the present invention. Administration
by inhalation is particularly useful in treating viral infections
of the lung, such as influenza. The aerosol can be administered
through the respiratory system or nasal passages. For example, one
skilled in the art will recognize that a composition of the present
invention can be suspended or dissolved in an appropriate carrier,
e.g., a pharmaceutically acceptable propellant, and administered
directly into the lungs using a nasal spray or inhalant. For
example, an aerosol formulation comprising a viral inducing agent
and/or antiviral agent can be dissolved, suspended or emulsified in
a propellant or a mixture of solvent and propellant, e.g., for
administration as a nasal spray or inhalant. Aerosol formulations
may contain any acceptable propellant under pressure, such as a
cosmetically or dermatologically or pharmaceutically acceptable
propellant, as conventionally used in the art.
[0194] An aerosol formulation for nasal administration is generally
an aqueous solution designed to be administered to the nasal
passages in drops or sprays. Nasal solutions can be similar to
nasal secretions in that they are generally isotonic and slightly
buffered to maintain a pH of about 5.5 to about 6.5, although pH
values outside of this range can additionally be used.
Antimicrobial agents or preservatives can also be included in the
formulation.
[0195] An aerosol formulation for inhalations and inhalants can be
designed so that an agent or combination of agents can be carried
into the respiratory tree of the subject when administered by the
nasal or oral respiratory route. Inhalation solutions can be
administered, for example, by a nebulizer. Inhalations or
insufflations, comprising finely powdered or liquid drugs, can be
delivered to the respiratory system as a pharmaceutical aerosol of
a solution or suspension of the agent or combination of agents in a
propellant, e.g., to aid in disbursement. Propellants can be
liquefied gases, including halocarbons, for example, fluorocarbons
such as fluorinated chlorinated hydrocarbons,
hydrochlorofluorocarbons, and hydrochlorocarbons, as well as
hydrocarbons and hydrocarbon ethers.
[0196] Halocarbon propellants can include fluorocarbon propellants
in which all hydrogens are replaced with fluorine,
chlorofluorocarbon propellants in which all hydrogens are replaced
with chlorine and at least one fluorine, hydrogen-containing
fluorocarbon propellants, and hydrogen-containing
chlorofluorocarbon propellants. Halocarbon propellants are
described in Johnson, U.S. Pat. No. 5,376,359, issued Dec. 27,
1994; Byron et al., U.S. Pat. No. 5,190,029, issued Mar. 2, 1993;
and Purewal et al., U.S. Pat. No. 5,776,434, issued Jul. 7, 1998.
Hydrocarbon propellants useful in the invention include, for
example, propane, isobutane, n-butane, pentane, isopentane and
neopentane. A blend of hydrocarbons can also be used as a
propellant. Ether propellants include, for example, dimethyl ether
as well as the ethers. An aerosol formulation of the invention can
also comprise more than one propellant. For example, an aerosol
formulation can comprise more than one propellant from the same
class, such as two or more fluorocarbons; or more than one, more
than two, more than three propellants from different classes, such
as a fluorohydrocarbon and a hydrocarbon. Pharmaceutical
compositions of the present invention can also be dispensed with a
compressed gas, e.g., an inert gas such as carbon dioxide, nitrous
oxide or nitrogen.
[0197] Aerosol formulations can also include other components, for
example, ethanol, isopropanol, propylene glycol, as well as
surfactants or other components such as oils and detergents. These
components can serve to stabilize the formulation and/or lubricate
valve components.
[0198] The aerosol formulation can be packaged under pressure and
can be formulated as an aerosol using solutions, suspensions,
emulsions, powders and semisolid preparations. For example, a
solution aerosol formulation can comprise a solution of an agent,
such as a viral inducing agent and/or antiviral agent in
(substantially) pure propellant or as a mixture of propellant and
solvent. The solvent can be used to dissolve the agent and/or
retard the evaporation of the propellant. Solvents useful in the
invention include, for example, water, ethanol and glycols. Any
combination of suitable solvents can be used, optionally combined
with preservatives, antioxidants, and/or other aerosol
components.
[0199] An aerosol formulation can also be a dispersion or
suspension. A suspension aerosol formulation may comprise a
suspension of an agent or combination of agents of the instant
invention, e.g., a viral inducing agent and/or antiviral agent, and
a dispersing agent. Dispersing agents useful in the invention
include, for example, sorbitan trioleate, oleyl alcohol, oleic
acid, lecithin and corn oil. A suspension aerosol formulation can
also include lubricants, preservatives, antioxidant, and/or other
aerosol components.
[0200] An aerosol formulation can be formulated as an emulsion. An
emulsion aerosol formulation can include, for example, an alcohol
such as ethanol, a surfactant, water and a propellant, as well as
an agent or combination of agents, e.g., a viral inducing agent
and/or an antiviral agent. The surfactant used can be nonionic,
anionic or cationic. One example of an emulsion aerosol formulation
comprises, for example, ethanol, surfactant, water and propellant.
Another example of an emulsion aerosol formulation comprises, for
example, vegetable oil, glyceryl monostearate and propane.
[0201] Pharmaceutical compositions suitable for use in the present
invention can include compositions wherein the active ingredients
are present in an effective amount, i.e., in an amount effective to
achieve therapeutic and/or prophylactic benefit in a host with at
least one virus-induced inflammatory condition. The actual amount
effective for a particular application will depend on the condition
or conditions being treated, the condition of the subject, the
formulation, and the route of administration, as well as other
factors known to those of skill in the art. Determination of an
effective amount of a viral inducing agent and/or antiviral agent
is well within the capabilities of those skilled in the art, in
light of the disclosure herein, and can be determined using routine
optimization techniques.
[0202] An effective amount for use in humans can be determined from
animal models. For example, a dose for humans can be formulated to
achieve circulating, liver, topical and/or gastrointestinal
concentrations that have been found to be effective in animals. One
skilled in the art can determine the effective amount for human
use, especially in light of the animal model experimental data
described herein. Based on animal data, and other types of similar
data, those skilled in the art can determine an effective amount of
a composition appropriate for humans.
[0203] An effective amount when referring to an agent or
combination of agents of the invention can generally mean the dose
ranges, modes of administration, formulations, etc., that have been
recommended or approved by any of the various regulatory or
advisory organizations in the medical or pharmaceutical arts (e.g.,
FDA, AMA) or by the manufacturer or supplier.
[0204] Further, appropriate doses for a viral inducing agent and/or
antiviral agent can be determined based on in vitro experimental
results.
[0205] A person of skill in the art would be able to monitor in a
patient the effect of administration of a particular agent. For
example, HIV or EBV viral load levels can be determined by
techniques standard in the art, such as measuring CD4 cell counts,
and/or viral levels as detected by PCR. Other techniques would be
apparent to one of skill in the art.
Administration Schedule
[0206] Administration of one or more agents (e.g, a viral inducing
agent and/or an antiviral) can be intermittent; for example,
administration can be once every two days, every three days, every
five days, once a week, once or twice a month, and the like. The
amount, forms, and/or amounts of the different forms can be varied
at different times of administration.
[0207] Pulsed administration of one or more pharmaceutical
compositions can be used for the treatment or prevention of a
viral-induced inflammatory condition. Pulsed administration can be
more effective than continuous treatment as pulsed doses can be
lower than would be expected from continuous administration of the
same composition. Each pulse dose can be reduced and the total
amount of drug administered over the course of treatment to the
patient can be minimized.
[0208] With pulse therapy, in vivo levels of an agent can drop
below that level required for effective continuous treatment.
Pulsed administration can reduce the amount of the composition
administered to the patient per dose or per total treatment regimen
with an increased effectiveness. Pulsed administration can provide
a saving in time, effort and expense and a lower effective dose can
lessen the number and severity of complications that can be
experienced by a subject. As such, pulsing can be more effective
than continuous administration of the same composition.
[0209] Individual pulses can be delivered to a subject continuously
over a period of several hours, such as about 2, 4, 6, 8, 10, 12,
14 or 16 hours, or several days, such as 2, 3, 4, 5, 6, or 7 days,
or from about 1 hour to about 24 hours or from about 3 hours to
about 9 hours. Alternatively, periodic doses can be administered in
a single bolus or a small number of injections of the composition
over a short period of time, for example, less than 1 or 2 hours.
For example, arginine butyrate can be administered over a period of
4 days with infusions for about 8 hours per day or overnight,
followed by a period of 7 days of no treatment.
[0210] The interval between pulses or the interval of no delivery
can be greater than 24 hours or can be greater than 48 hours, and
can be for even longer such as for 3, 4, 5, 6, 7, 8, 9 or 10 days,
two, three or four weeks or even longer. The interval between
pulses can be determined by one of ordinary skill in the art. The
interval between pulses can be calculated by administering another
dose of the composition when the composition or the active
component of the composition is no longer detectable in the patient
prior to delivery of the next pulse. Intervals can also be
calculated from the in vivo half-life of the composition. Intervals
can be calculated as greater than the in vivo half-life, or 2, 3,
4, 5 and even 10 times greater than the composition half-life.
Intervals can be 25, 50, 100, 150, 200, 250 300 and even 500 times
the half life of the chemical composition.
[0211] The number of pulses in a single therapeutic regimen can be
as little as two, but can be from about 5 to 10, 10 to 20, 15 to 30
or more. Subjects (e.g., patients) can receive one or more agents
(e.g., drugs) for life according to the methods of this invention.
Compositions can be administered by most any means, and can be
delivered to the patient as an injection (e.g. intravenous,
subcutaneous, intraarterial), infusion or instillation, and more
preferably by oral ingestion. Various methods and apparatus for
pulsing compositions by infusion or other forms of delivery to the
patient are disclosed in U.S. Pat. Nos. 4,747,825; 4,723,958;
4,948,592; 4,965,251 and 5,403,590.
[0212] In certain embodiments, the co-formulated unit dose
comprising an HDAC inhibitor and an antiviral agent is administered
daily. In further embodiments, administration is continuous. In
some embodiments, the administration of the co-formulated unit dose
is by pulsed administration. In certain embodiments, pulsed
administration comprises administering pulses of the co-formulated
unit dose for about 1 day, about 2 days, about 3 days, about 4
days, about 5 days, about 6 days, about 7 days, about 10 days,
about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6
weeks, about 7 weeks, about 8 weeks, about 2 months, about 3
months, about 4 months, about 5 months, about 6 months, about 9
months, about 12 months. In some embodiments, pulsed administration
comprises intervals of not administering the co-formulated unit
dose of about 1 day, about 2 days, about 3 days, about 4 days,
about 5 days, about 6 days, about 7 days, about 10 days, about 2
weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks,
about 7 weeks, about 8 weeks, about 2 months, about 3 months, about
4 months, about 5 months, about 6 months, about 9 months, about 12
months.
[0213] In some embodiments, the administration of the co-formulated
unit dose is by pulsed administration. In certain embodiments, the
pulsed administration comprises administering the co-formulated
unit dose for about 8 weeks, followed by not administering the
co-formulated unit dose for about 4 weeks. In some embodiments, the
pulsed administration comprises administering the co-formulated
unit dose for about 6 weeks, followed by not administering the
co-formulated unit dose for about 2 weeks. In certain embodiments,
the pulsed administration comprises administering the co-formulated
unit dose for about 4 weeks, followed by not administering the
co-formulated unit dose for about 2 weeks. In some embodiments, the
pulsed administration comprises administering the co-formulated
unit dose for about 2 weeks, followed by not administering the
co-formulated unit dose for about 2 weeks. In some embodiments,
pulsed administration comprises pulses of administering the
co-formulated unit dose for about 1 day, about 2 days, about 3
days, about 4 days, about 5 days, about 6 days, about 7 days, about
10 days, about 2 weeks, about 3 weeks, about 4 weeks, about 5
weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 2 months,
about 3 months, about 4 months, about 5 months, about 6 months,
about 9 months, about 12 months. In certain embodiments, pulsed
administration comprises intervals of not administering the
co-formulated unit dose of about 1 day, about 2 days, about 3 days,
about 4 days, about 5 days, about 6 days, about 7 days, about 10
days, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks,
about 6 weeks, about 7 weeks, about 8 weeks, about 2 months, about
3 months, about 4 months, about 5 months, about 6 months, about 9
months, about 12 months. In some embodiments, administration is
continuous. In certain embodiments, administration is for the
lifetime of the subject. In other embodiments, administration is
for about 1 week, about 2 weeks, about 3 weeks, about 4 weeks,
about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 2
months, about 3 months, about 4 months, about 5 months, about 6
months, about 9 months, or about 12 months. In some embodiments, an
antiviral agent is administered during intervals of not
administering the co-formulated unit dose. In certain embodiments,
an antiviral agent is administered in addition to the co-formulated
unit dose. In some embodiments, an antiviral agent is administered
simultaneously with the co-formulated unit dose. In other
embodiments, an antiviral agent is administered separate from the
co-formulated unit dose.
[0214] A pharmaceutical composition comprising a viral inducing
agent can be administered to a subject before a pharmaceutical
composition comprising an antiviral agent is administered to the
subject. A pharmaceutical composition comprising a viral inducing
agent can be co-administered to a subject with a pharmaceutical
composition comprising an antiviral agent. A pharmaceutical
composition comprising a viral inducing agent can be
co-administered with a pharmaceutical composition comprising an
antiviral agent and a pharmaceutical composition comprising one or
more addition agents. The pharmaceutical compositions can be
provided by pulsed administration. For example, a pharmaceutical
composition comprising a viral inducing agent can be administered
to a subject, followed by administration of a pharmaceutical
composition comprising an antiviral agent to the subject after an
interval of time has passed, and this order of administration the
same or similar time interval can be repeated, for example, at
least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more times.
EXAMPLES
Example 1
Phase 1 Study of AB Plus Ganciclovir in Patients with EBV
Associated Lymphoid Malignancies
[0215] Fifteen patients with EBV-associated lymphoid malignancies,
who had histologically confirmed lymphoid neoplasms that were EBV+,
were treated with AB and GCV. Prior therapies (varying in different
subjects) included rituximab, chemotherapy, chemoradiotherapy and
bone marrow transplant. GCV was administered at a rate of 5 mg/kg
intravenously (IV) over 1 hour twice per day, and continued
throughout the cycle. AB was continuously infused at a starting
dose of 500 mg/kg/day. Dose escalation was continued as follows
until MTD was established:
Level 1: 500 mg/kg/day IV for 2 days Level 2: 1000 mg/kg/day IV for
2 days Level 3: 1500 mg/kg/day IV for 2 days Level 4: 2000
mg/kg/day IV until day 21
[0216] A total of 15 patients were evaluated for anti-tumor
response (Table 1). A complete response (CR) was defined as
disappearance of detectable malignant disease on imaging or
physical examination (e.g., for skin lesions or tonsillar masses).
A partial response (PR) was defined as a 50% decrease in tumor size
(the sum of the product of the largest perpendicular diameters) or
measurable lesions chosen for analysis prior to beginning of
treatment. For lesions which could only be measured in 1 dimension,
such as skin (cutaneous T cell lymphoma), a greater than 50%
decrease in the largest dimension qualified as a PR. For the 3
patients who died from co-morbidities, anti-tumor responses were
confirmed pathologically at autopsy.
TABLE-US-00001 TABLE 1 Treatment Courses in Patients with EBV
Associated Lymphoid Malignancies Treated with AB and GCV Patient
HD/HTD.sup.1 Outcome, Number No. Cycles (mg/kg/day) 1 cycle Adverse
Events 1 <1, 15 d 500 CR confusion; diarrhea; emesis; rejection
of lung transplant* 2 <1, 16 d 1800 CR confusion; mucositis;
headache; nausea; vomiting; abdominal pain 3 <1, 19 d 2000 PR
confusion; mucositis; headache; nausea/vomiting; tumor regression
preceding bowel perforation* 4 1 2000 PR confusion:
nausea/vomiting; anorexia 5 1 2000 NR confusion; restlessness;
somnolence; nausea; vomiting; abdominal pain; vision change;
orthostasis 6 1 1000/1000 NR Headache; nausea/vomiting; abdominal
pain; thrombocytopenia 7 1 2000/1500 CR Lethargy/stupor/confusion;
hypotonia/hypoesthesia; fungal infection/mucositis; tumor lysis
leading to hemorrhage* 8 1 1500/1000 PR Acoustic hallucinations;
somnolence; hypokalemia; sepsis; Deep Vein Thrombosis 9 1 2000/2000
CR Confusion; fatigue; elevated BUN; tumor lysis leading to
pancreatitis/hepatitis* 10 1 1000/800 NR Elevated BUN,
encephalopathy 11 <1, 8 d 1500/1500 PR Diarrhea; hepatomegaly 12
1 2000/2000 NR Nausea; pneumonia; port infection 13 3 938/938 PR
Nausea; anorexia; weight loss; anemia; thrombocytopenia; lethargy;
insomnia; hypokalemia 14 <1, 19 d 1250/1250 NR Sinus, throat,
back pain; thrombocytopenia; hypokalemia; lethargy 15 <1, 5 d
1000/1000 PR Lethargy; increased dyspnea; polymicrobial
pneumonia/acute respiratory distress syndrome *fatal AE;
.sup.1HD/HTD-highest dose/highest tolerated dose.
[0217] Four patients were classified as achieving CRs, including 2
with PTLD, 1 with extranodal NK/T cell lymphoma and 1 with
peripheral T-cell lymphoma. Three of these patients died after
completing therapy as a result of co-morbid conditions and
complications presumed related to tumor regression. Autopsy
examination of 2 subjects revealed apparent complete disappearance
of tumor, while the third patient demonstrated significant necrosis
of residual lymphoma at autopsy.
[0218] Six patients were classified as partial responses (PRs),
including 3 with PTLD, 1 with diffuse large-cell B-cell lymphoma, 1
with extranodal NK/T-cell lymphoma and 1 with subcutaneous
panniculitis-like T-cell lymphoma.
[0219] The remaining 5 patients were classified as non-responders
(NR).
[0220] In summary 10 out of 15 patients showed a degree of response
to treatment of AB in combination with the antiviral
ganciclovir.
Example 2
Phase II Trial of Low-Dose Arginine Butyrate and
Ganciclovir/Valganciclovir in EBV(+) Lymphoid Malignancies
[0221] It has previously been found that continuous infusion of
inducing agent, for example, arginine butyrate, may not be
necessary to maintain viral thymidine kinase expression and
sensitization to anti-viral agents in EBV-associated tumors, but
that, in fact, cells that survived initial exposure to the inducing
agent plus the anti-viral agent remained susceptible to further
cycles of combination treatment (Ghosh, S. K., et al. 2007 Blood
Cells, Molecules, and Diseases 38:57-65, incorporated herein in its
entirety). However, it was neither anticipated nor expected that
after some first period of treatment with inducing agent and
anti-viral agent, one could continue the anti-viral treatment
effectively within a cycle of therapy without continued
administration of the inducing agent (continued administration
including continued periods of pulsing throughout).
[0222] A clinical trial was instituted utilizing a 5-day infusion
of arginine butyrate and 21 days of ganciclovir/valganciclovir for
EBV+ lymphomas and Post-transplant Lymphoproliferative Disorder
(PTLD). The first patient enrolled in the protocol (with
Rituximab-refractory PTLD following a cord stem cell
transplantation for Hodgkin Disease) tolerated the treatment
regimen well, with resolution of cough within three days and a
decrease in LDH levels.
[0223] Treatment with arginine butyrate (AB) was administered in a
hospital/inpatient basis. The subject was a 32 year old with
EBV-related post-transplant lymphoma who had failed multiple
therapies (chemotherapy, Rituxan). The subject received AB 1,000
mg/kg/dose intravenously for 5 days (day 1-5). The dose was given
continuously over 24 hours. AB was given through a long IV line or
port due to hypertonicity. Ganciclovir at 5 mg/kg IV over 1 hour
was given twice a day for five days (day 1-5). Valganciclovir 900
mg was given orally twice per day for 16 days (day 6-21). At the
end of the 21-day cycle, imaging studies were done to determine
response and revealed elimination of nearly all tumor masses (FIG.
1). Four of six target lesions resolved completely, and two
additional lesions decreased in size. (Table 2) The subject's
symptoms of fever and cough resolved for first time in 9 weeks.
Measure of the tumor marker serum LDH was reduced from 899 to 328
(normal). Additionally, EBV, CMV, and HH6 viral load became
undetectable. These findings indicate that a shorter, more
patient-accessible regimen of the virus-target therapeutic strategy
is more efficacious. Also, there remains a continuous need for an
oral as opposed to an intravenous HDAC inhibitor. Therefore, the
present invention contemplated oral HDAC inhibitors having
increased potency as compared to AB.
TABLE-US-00002 TABLE 2 Quantificationof tumor response evaluated by
CT Scan. Tumor dimensions in cm. Pre-Treatment Post-Treatment
Dimension Dimension Dimension Dimension Location 1 2 1 2 R. Upper
lobe 0.7 0.7 None None R. Mid Lobe 1.1 1.1 None None R. Lower Lobe
1.4 0.8 0.8 0.6 L. Upper Lobe 0.9 0.8 None None L. Lower Lobe 0.9
0.6 0.6 0.5 Lingular 0.9 0.7 None None Hepatic Seg. 6 1.1 1.1 None
None Hepatic Seg. 8 1.0 0.7 None None L. Ant. Abd. Wall 1.1 1.9
None None R. Ant. Abd. Wall 0.9 0.5 0.9 0.5
Example 3
Analysis of Efficacy of the Herpes Anti-Virals
[0224] There are 12 mammalian HDACs, and any one of which might be
required for repression of the TK or EBV-PK gene during latency in
tumors. HDAC isozyme-specific siRNAs were used to knockdown
individual HDACs in tumor lines expressing latent EBV to determine
which one of them induces reactivation of TK from latency,
rendering it susceptible to anti-virals.
[0225] The EBV-positive B lymphoma cell line P3HR1 was used
throughout these assays. The P3HR1 cell line was originally derived
from Burkitt's lymphoma patient. EBV maintains a latent state of
replication in this cell line. Cells were maintained in RPMI 1640
with 10% fetal bovine serum containing 100 U penicillin per ml and
100 .mu.g streptomycin per ml. The HDAC inhibitors used were from
five different classes: a) short chain fatty acids, b) hydroxamic
acids, c) benzamides, d) cyclic tetrapeptides, and e)
largazoles.
[0226] To measure the relative level of TK mRNA in various total
RNA preparations, reverse transcription and quantitative PCR using
real time PCR technology was used. Five micrograms of total RNA was
reverse-transcribed using random hexamer primers and Superscript
III cDNA synthesis kit (Invitrogen). The cDNA was diluted to a
final volume of 60 .mu.l with sterile water, 8 .mu.l of which was
then used in each real time PCR reaction in an ABI 7500 Sequencher
using SYBR-Green technology. Primers used for the amplification of
TK were EBV-TK1-F: 5'-AGATGACGACGGCCTCTACCA-3' (SEQ ID NO: 1);
EBV-TK1-R: 5'-CCTCCTTCTGTGCACGAAGT-3' (SEQ ID NO: 2). The
.beta.-actin mRNA levels in those samples were determined similarly
using .beta.-actin-specific primers Actin/hu-F:
5'-GCTCGTCGTCGACAACGGCTC-3' (SEQ ID NO: 3); Actin/hu-R:
5'-CAAACATGATCTGGGTCATCTTCTC-3' (SEQ ID NO: 4). The relative level
of TK expression in a sample was calculated following normalization
of .beta.-actin expression level.
[0227] Toxicity assays with two anti-herpesvirus drugs, Gancicovir
(GCV) and Penciclovir (PCV), treated to P3HR1 cells alone was
conducted as a control. A total of 3.times.10.sup.5 P3HR1 cells
were incubated with various concentrations of GCV or PCV and
incubated for 6 days. Viable cell counts were measured and toxicity
was expressed as percentage of cell growth compared to untreated
cells. As shown in FIG. 2A and FIG. 2B, PCV was less toxic to the
cells compared to GCV. The effect of 40 .mu.M GCV and PCV in
combination treatment approach with 1.0 mM Na-butyrate in P3HR1
cells was compared (FIG. 2C) Inhibition of cell growth with 40
.mu.M PCV (76%) was much less than with 40 .mu.M GCV (38%). This
lower level of inhibition of cell growth with PCV did not change
significantly when the drug was used at higher concentrations (FIG.
2D).
Example 4
Analysis of Efficacy of HDAC Inhibitors
[0228] The induction of lytic phase was assayed in EBV-positive
lymphoma cell lines exposed to different HDAC inhibitors (HDACi)
for 24-48 hrs, then the expression of EBV TK and other EBV
transcripts by RT-PCR analysis was quantified. To determine tumor
cytotoxic activity of the combination of HDACi and GCV, EBV+
lymphoma cells were exposed to a range of concentrations of HDAC
inhibitors and ganciclovir (GCV) for 3 days and then to GCV alone
for another 3 days. Efficacy of a particular HDAC inhibitor in the
combination treatment approach was then determined by enumerating
living cells. A general experimental protocol is described
below.
Cells
[0229] The EBV-positive B lymphoma cell line P3HR1 was used in the
study. The P3HR1 cell line was originally derived from Burkitt's
lymphoma patient. EBV maintains a latent state of replication in
this cell line. These cells were maintained in RPMI 1640 with 10%
fetal bovine serum containing 100 U penicillin per ml and 100 .mu.g
streptomycin per ml.
Study Agent
[0230] Various HDAC inhibitors were evaluated in this study. As a
positive control, the Short Chain Fatty Acid butyrate, an
established inducer of EBV-TK was used. Ganciclovir (GCV) was used
as the anti-viral drug.
Titration of HDAC Inhibitors on P3HR1 Cells:
[0231] Concentrations of HDAC inhibitors, which do not
significantly affect the viability or proliferation of P3HR1 cells
in culture were established.
Drug Sensitivity Assay
[0232] To test the sensitivity of EBV-positive lymphoma cells
towards HDAC inhibitors, P3HR1 cells were treated with HDAC
inhibitors in the presence of one anti-viral drug. At the end of
the assay, the efficacy of the HDAC inhibitors was assessed by
measuring the inhibition of cell growth compared to untreated
cells.
[0233] Healthy, actively-growing P3HR1 cells were harvested and
resuspended in fresh growth media. Cells were seeded in wells.
Appropriate dilutions of HDAC inhibitors were added to certain
wells, some of which received an anti-viral drug (such as GCV at 50
.mu.M concentration). At 72 hrs, 800 .mu.l of culture fluid was
removed from each well. Wells were then refed with 1.0 ml fresh
growth media without HDAC inhibitors. Fresh GCV solution was added
to the wells that originally received GCV at the same initial
concentration. On day 6, cell morphology was observed under
microscope and viable cell counts in each individual wells were
determined by trypan blue dye exclusion method using an automated
cell counter (Countess, Invitrogen).
Thymidine Kinase Expression Assay
[0234] A Thymidine Kinase (TK) was used to determine if HDAC
inhibitors induced TK expression in EBV-infected lymphoma
cells.
[0235] P3HR1 cells were seeded in 60 mm plates containing
3.times.106 cells in 3 ml of fresh growth media. Appropriate
concentrations of HDAC inhibitors were added to the plate and cells
were incubated in the presence of HDAC inhibitors for 6 h, 24 h, 48
h, or as needed. Cells were harvested by centrifugation and washed
once in cold PBS. Total cellular RNA was then extracted. To measure
the relative level of TK mRNA in various total RNA preparations,
reverse transcription and quantitative PCR using real time PCR
analysis were used. See, Ghosh, S. K., Forman, L. W., Akinsheye,
I., Perrine, S. P., Faller, D. V.: Short discontinuous sodium
butyrate exposure efficiently sensitizes latently EBV infected
cells towards nucleoside analogue-mediated growth inhibition, Blood
Cells Mol. Diseases (2007), 38:57-65. The relative level of TK
expression in a sample was calculated following normalization of
.beta.-actin expression level.
Results
[0236] The HDAC inhibitors had varying levels of synergistic
activity with anti-viral agents in killing EBV+ lymphoma cells. The
hydroxamic acid LBH589, the benzamide MS275, and synthetic
largazole derivatives were 10.sup.4 to 10.sup.5-times more potent
in killing EBV+ lymphoma cells in the presence of GCV, compared to
sodium butyrate. The effective concentration of LBH589 was in the
range of 50-100 nM, MS275 at 200-500 nM, and Largazole derivatives
at 100-200 nM. At these concentrations, the drugs as single agents
produced no significant growth inhibitory activity in the tumor
cells. LBH589, MS275 and Largazole derivatives also strongly
induced EBV-TK expression in the tumor cells. In certain instances,
the effectiveness of these HDACi compounds at such low
concentrations makes them potentially applicable as sensitizers to
anti-viral therapeutics for the treatment of EBV-associated
lymphomas and other viral or virally-induced conditions. In some
embodiments of the invention, these HDAC inhibitors are used as an
alternative therapeutic option, in combination with antivirals, for
the treatment of EBV-associated tumors and other viral or
virally-induced conditions.
A. Short Chain Fatty Acids
[0237] Two SCFA HDAC inhibitors, Na-butyrate (NaB) and valproic
acid (VA) were tested.
[0238] Sodium Butyrate (NaB):
[0239] In a combination treatment approach, NaB+GCV reduced growth
of EBV-positive P3HR1 cells significantly (up to 50% more) compared
to cells treated with NaB or GCV alone (FIG. 3A). The optimal
concentration of NaB for this purpose was found to be 1.0 mM.
Responses from 1.0 mM NaB was used as a control for interpreting
results in this experiment. At a higher concentration, NaB alone
reduces cell growth to a significant degree, and the synergistic
effects of GCV are lost at those concentrations of NaB.
[0240] Valproic Acid (VA):
[0241] The other HDACi used in this experiment, VA, also had very
similar activity (FIG. 3B). Analysis of TK mRNA level by RT and
real-time PCR however showed that VA was less efficient than NaB in
inducing TK expression (FIG. 3C).
B. Hydroxamic Acids
[0242] A total of five different HDAC inhibitors from the
hydroxamic acid group were examined as combination therapies. These
inhibitors include scriptaid, SAHA, panobinostat-LHB589,
belinostat-PXD101, and oxamflatin. All of these HDAC inhibitors can
be administered orally via an oral formulation.
[0243] Scriptaid:
[0244] Scriptaid showed strong synergistic effect with GCV in
reducing cell growth of P3HR1 cells, especially at 500 nM and 1
.mu.M concentrations (FIG. 4A). As shown in the data, in preferred
embodiments, an HDAC inhibitor of the invention combined with an
antiviral agent can reduce the cell count of EBV infected cells to
less than 70%, 60%, 50%, 40%, 30%, 20%, or 10% of cells treated
without the HDAC inhibitor or antiviral treatment, or by at least
60%, 50%, 40%, 30%, or 20% from cells treated with an antiviral
agent alone.
[0245] SAHA-Vorinostat:
[0246] The combination treatment experiment with SAHA showed that
it could induce TK expression at a higher level than that seen with
efficient concentrations of butyrate (1.0 mM) (FIG. 5B). As such,
the present invention contemplates using an HDAC inhibitor
(preferably an oral HDAC inhibitor) to induce EBV kinase expression
by at least 2-, 4-, 6-, 8-, or 10-fold, wherein the HDAC inhibitor
is administered at a concentration of less than 0.5 mg/kg, 1 mg/kg,
2 mg/kg, 4 mg/kg, 6 mg/kg, 8 mg/kg, or 10 mg/kg.
[0247] LHB589-Panobinostat:
[0248] The growth inhibitory activity of LHB589 at a 50 nM
concentration was comparable to that of NaB at 1.0 mM (FIG. 6A).
When the cells were treated for 3 days or longer, LHB589 was
extremely toxic to the cells at any concentrations 100 nM or above.
Although when treated for 24 h only, cells survived well even at a
concentration of 5 .mu.M. TK expression level in presence of LHB589
was quite high compared to optimum concentration of NaB (2.5 mM)
(FIG. 6B). Thus, in some embodiments, the present invention
contemplates a short administration of an oral HDAC inhibitor
(e.g., less than 4 days, 3 days, 2 days, 36 h, 24 h, 12 h, or 6 h)
in combination with or followed by an antiviral treatment.
[0249] PXD101-Belinostat:
[0250] PXD101 induced high level of TK expression at the 5 .mu.M
concentration. (FIG. 7). Thus, the present invention contemplates
using an HDAC inhibitor (preferably an oral HDAC inhibitor) to
induce EBV kinase expression by at least 5-, 10-, 15-, 20-, 25-,
30-, or 35-fold.
[0251] Oxamflatin:
[0252] Oxamflatin showed synergistic activity with GCV towards
reducing cell growth. At a 200 nM concentration, the activity level
(growth suppression) was more than what typically seen with 1.0 mM
NaB (FIG. 8).
[0253] As shown in the data, hydroxamic acid HDAC inhibitors
synergistically with GCV reduce cell growth of P3HR1 cells.
Furthermore, hydroxamic acid HDAC inhibitors induce EBV kinase
expression by at least 2-, 5-, 10-, 20-, or 30-fold. In addition,
hydroxamic acid HDAC inhibitors in combination with antiviral
agents reduce the cell count of EBV infected cells to less than
70%, 60%, 50%, 40%, 30%, 20%, or 10% of cells treated without the
HDAC inhibitor or antiviral treatment.
C. Cyclic Tetrapeptide
[0254] Apicidin:
[0255] The cyclic tetrapeptide group of HDACi examined was
apicidin. A toxicity assay with apicidin alone showed that
concentrations of apicidin higher than 200 nM was quite toxic to
the cells. The combination treatment assay (FIG. 9) showed that at
100 nM and 200 nM concentrations, apicidin reduced cell growth by
40-50% over cells treated with apicidin alone. However, the 200 nM
concentration cell growth was significantly retarded without any
GCV and a 500 nM concentration was very toxic to the cells.
D. Benzamide
[0256] MS-275:
[0257] Experiments show that the benzamide class of HDAC inhibitors
were extremely potent in sensitizing P3HR1 cells to GCV-mediated
effects. As shown below (FIG. 10A) a 500 nM concentration of MS-275
was as efficient as 1.0 mM NaB. Higher concentrations were
extremely toxic to the cells. Interestingly, MS-275 also strongly
induced TK expression at 500 nM and higher concentrations (FIG.
10B). TK expression was also induced at only 6 hr post treatment.
Based on these results, an even shorter exposure to MS-275 was
examined to see if it would be sufficient to sensitize P3HR1 cells
to GCV-mediated killing. Cells with were treated with MS-275 and
GCV for shorter time periods of 24 hr or 48 hr (as opposed to 72
hr) and then further incubated in presence of GCV for up to 6 days,
at which time the viable cell counts were enumerated. As shown in
FIG. 10C, even at just 24 hr exposure to MS-275 sensitized the
cells to GCV-mediated effects as efficiently as a 72 hr continuous
treatment. This further demonstrates that MS-275 is very effective
sensitizing agent for combination treatment studies.
E. Largazole
[0258] Largazole Derivatives:
[0259] Largazole is a member of macrocyclic depsipeptide that was
originally isolated from coral reef cyanobacteria. Largazole is a
potent HDAC inhibitor with specificity towards HDAC class 1 and 2
only. Additionally, largazole has very low IC 50 and HDAC isozyme
specificity. 16 different analogs of the largazole were tested
(ab6-113b, ab6-113a, ab6-123a, ab6-123b, ab6-164b, ab6-156b, 232a,
233a, 238a, 233b, 234b, 235b, 234a, 235a, 237a, 212b, TLN1 357,
TNL2 380, ART01) for synergistic cell killing activity in
combination with GCV (FIGS. 12A, B, C, D, and E). In some
embodiments, the HDAC inhibitor and antiviral treatment reduces the
number of EBV TK induced cells by at least 10%, 20%, 30%, 40%, 50%,
60%, 70%, 80%, or 90% as compared to untreated cells or cells
treated with the antiviral agent only. 13 largazole derivatives
were tested both in combination treatment approach and also for
their ability to induce EBV TK (FIG. 12F). Several of the
largazoles showed potent cell killing activity in combination with
GCV. Thus, in some embodiments, an HDAC inhibitor of the invention
is one that can induce EBV TK by at least 2-, 4-, 6-, 8-, 10-, 12-,
or 14-fold as compared to cells not treated with an HDAC inhibitor.
In any of the embodiments herein, the HDAC inhibitor is preferably
suitable for oral formulation.
Example 5
Analysis of Efficacy of Combination Treatment with HIV-Infected
Cells
[0260] Virus production (p24 release) was examined in an
HIV-1-infected monocyte line. Cells were treated or not treated
with HDAC-inhibitors and other compounds. P24 release expressed as
optical density ("OD") (FIG. 13), and then converted to pg of
protein (FIG. 14). Arginine butyrate, phorbol myristate acetate
(PMA), trichostatin A (TSA), LHB589, apicidin (API) and largazole
(LARG) are shown to be active, whereas 2,2-dimethyl butyrate (ST20)
and 2-(quinazolin-4-ylamino)butanoic acid (RB3) increased viral
production at levels similar to the control of vehicle alone. DMSO
was vehicle for some of the compounds tested.
Prophetic Example 1
Treatment of Multiple Sclerosis
[0261] To treat a subject suspected of having Epstein-Barr
virus-induced multiple sclerosis, a health care professional
administers to the subject a dose of a pharmaceutical composition
comprising a viral inducing agent, the HDAC inhibitor JNJ-26481585.
A week later, a health care professional administers to the subject
a dose of a pharmaceutical composition comprising an antiviral
agent, valganciclovir. A cycle of administration is carried out for
a period of a month (FIG. 15). During the period of administration
of the viral inducing agent and the antiviral agent, the subject is
also administered mitoxantrone to treat the multiple sclerosis. A
cycle of HDAC inhibitor and antiviral treatment may be repeated at
least 2, 4, 6, 8, 10, or 12 times as needed.
Prophetic Example 2
Treatment of Atherosclerosis
[0262] To treat a coronary artery condition patient suspected of
having cytomegalovirus-induced atherosclerosis, a health care
professional administers a dose of a pharmaceutical composition
comprising a viral inducing agent, JNJ-26481585. A week later, a
health care professional administers to the subject a dose of a
pharmaceutical composition comprising valganciclovir. A cycle of
administration is carried out for a period of a month. During the
period of administration of the viral inducing agent and the
antiviral agent, the subject is also administered rosuvastatin to
treat the atherosclerosis.
Prophetic Example 3
[0263] A patient either diagnosed with or suspected of having an
Epstein Barr Virus (EBV)-associated malignancy such as
nasopharyngeal carcinoma, Hodgkin's disease, Burkitt's lymphoma,
post-transplantation lymphoproliferative disease, or gastric
carcinoma can be treated. A health care professional administers a
dose of a pharmaceutical composition comprising JNJ-26481585 and
valganciclovir co-formulated for oral administration. The patient
is administered a single daily dose in tablet form, where the
tablet contains 5 mg of JNJ-26481585 and 1500 mg of timed-release
or slow-release valganciclovir. A cycle of administration is
carried out for a period of 28 days. During the period of
administration, the subject can optionally also be administered an
additional chemotherapeutic agent to treat the malignancy.
[0264] While preferred embodiments of the present invention have
been shown and described herein, it will be obvious to those
skilled in the art that such embodiments are provided by way of
example only. Numerous variations, changes, and substitutions will
now occur to those skilled in the art without departing from the
invention. It should be understood that various alternatives to the
embodiments of the invention described herein may be employed in
practicing the invention. It is intended that the following claims
define the scope of the invention and that methods and structures
within the scope of these claims and their equivalents be covered
thereby.
Sequence CWU 1
1
4121DNAArtificial SequenceDescription of Artificial Sequence
Synthetic primer 1agatgacgac ggcctctacc a 21220DNAArtificial
SequenceDescription of Artificial Sequence Synthetic primer
2cctccttctg tgcacgaagt 20321DNAArtificial SequenceDescription of
Artificial Sequence Synthetic primer 3gctcgtcgtc gacaacggct c
21425DNAArtificial SequenceDescription of Artificial Sequence
Synthetic primer 4caaacatgat ctgggtcatc ttctc 25
* * * * *