U.S. patent application number 16/466957 was filed with the patent office on 2021-11-18 for compounds and methods for reducing fxi expression.
This patent application is currently assigned to Ionis Pharmaceuticals, Inc.. The applicant listed for this patent is Ionis Pharmaceuticals, Inc.. Invention is credited to Huynh-Hoa Bui.
Application Number | 20210355497 16/466957 |
Document ID | / |
Family ID | 1000005799792 |
Filed Date | 2021-11-18 |
United States Patent
Application |
20210355497 |
Kind Code |
A1 |
Bui; Huynh-Hoa |
November 18, 2021 |
COMPOUNDS AND METHODS FOR REDUCING FXI EXPRESSION
Abstract
Provided are compounds, methods, and pharmaceutical compositions
for reducing the amount or activity of FXI RNA in a cell or
subject, and in certain instances reducing the amount of FXI
protein in a cell or subject. Such compounds, methods, and
pharmaceutical compositions are useful to prevent, treat, or
ameliorate at least one symptom of a thromboembolic condition
without a significant increase in a bleeding risk. Such
thromboembolic conditions include deep vein thrombosis, venous or
arterial thrombosis, pulmonary embolism, myocardial infarction,
stroke, thrombosis associated with chronic kidney disease or
end-stage renal disease (ESRD), including thrombosis associated
with dialysis, or other procoagulant condition. Such symptoms
include decreased blood flow through an affected vessel, death of
tissue, and death.
Inventors: |
Bui; Huynh-Hoa; (San Diego,
CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Ionis Pharmaceuticals, Inc. |
Carlsbad |
CA |
US |
|
|
Assignee: |
Ionis Pharmaceuticals, Inc.
Carlsbad
CA
|
Family ID: |
1000005799792 |
Appl. No.: |
16/466957 |
Filed: |
May 8, 2019 |
PCT Filed: |
May 8, 2019 |
PCT NO: |
PCT/US2019/031277 |
371 Date: |
June 5, 2019 |
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C12N 2310/3341 20130101;
C12N 15/1137 20130101; C12N 2320/30 20130101; C12N 2310/11
20130101; C12N 2310/321 20130101; C12N 2310/341 20130101; C12N
2310/322 20130101; C12N 2310/315 20130101 |
International
Class: |
C12N 15/113 20060101
C12N015/113 |
Claims
1. An oligomeric compound according to the following chemical
structure: ##STR00022## or a salt thereof.
2. An oligomeric compound according to the following chemical
structure: ##STR00023##
3. (canceled)
4. An oligomeric compound according to the following formula:
(THA-GalNAc.sub.3)o Aes mCeo Geo Geo mCeo Ads Tds Tds Gds Gds Tds
Gds mCds Ads mCds Aeo Geo Tes Tes Te (SEQ ID NO: 3); wherein,
(THA-GalNAc.sub.3)o is represented by the following chemical
structure: ##STR00024## and wherein, A=an adenine nucleobase, mC=a
5'-methyl cytosine nucleobase, G=a guanine nucleobase, T=a thymine
nucleobase, e=a 2'-MOE modified sugar, d=a 2'-deoxyribose sugar,
s=a phosphorothioate internucleoside linkage, and o=a
phosphodiester internucleoside linkage; or a salt thereof.
5. The oligomeric compound of claim 1, which is a sodium salt.
6. A chirally enriched population of the oligomeric compound of
claim 1, wherein the population is enriched for oligomeric
compounds having a modified oligonucleotide comprising at least one
particular phosphorothioate internucleoside linkage having a
particular stereochemical configuration.
7. The chirally enriched population of claim 6, wherein the
population is enriched for oligomeric compounds having a modified
oligonucleotide comprising at least one particular phosphorothioate
internucleoside linkage having the (Sp) configuration.
8. The chirally enriched population of claim 6, wherein the
population is enriched for oligomeric compounds having a modified
oligonucleotides comprising at least one particular
phosphorothioate internucleoside linkage having the (Rp)
configuration.
9. The chirally enriched population of claim 6, wherein the
population is enriched for oligomeric compounds having a modified
oligonucleotide having a particular, independently selected
stereochemical configuration at each phosphorothioate
internucleoside linkage.
10. The chirally enriched population of claim 9, wherein the
population is enriched for oligomeric compounds having a modified
oligonucleotide having the (Sp) configuration at each
phosphorothioate internucleoside linkage.
11. The chirally enriched population of claim 9, wherein the
population is enriched for oligomeric compounds having a modified
oligonucleotide having the (Rp) configuration at each
phosphorothioate internucleoside linkage.
12. The chirally enriched population of claim 9, wherein the
population is enriched for oligomeric compounds having a modified
oligonucleotide having at least 3 contiguous phosphorothioate
internucleoside linkages in the Sp-Sp-Rp configurations, in the 5'
to 3' direction.
13. A population of oligomeric compounds having a modified
oligonucleotide of claim 1, wherein all of the phosphorothioate
internucleoside linkages of the modified oligonucleotide are
stereorandom.
14. A pharmaceutical composition comprising the oligomeric compound
of claim 1, and a pharmaceutically acceptable carrier or
diluent.
15. The pharmaceutical composition of claim 14, wherein the
pharmaceutically acceptable diluent is phosphate buffered
saline.
16. The pharmaceutical composition of claim 14, wherein the
pharmaceutical composition consists or consists essentially of the
oligomeric compound and phosphate buffered saline.
17-63. (canceled)
64. A pharmaceutical composition comprising the oligomeric compound
of claim 2, and a pharmaceutically acceptable carrier or
diluent.
65. The pharmaceutical composition of claim 64, wherein the
pharmaceutically acceptable diluent is phosphate buffered
saline.
66. The pharmaceutical composition of claim 64, wherein the
pharmaceutical composition consists or consists essentially of the
oligomeric compound and phosphate buffered saline.
Description
SEQUENCE LISTING
[0001] The present application is being filed along with a Sequence
Listing in electronic format. The Sequence Listing is provided as a
file entitled BIOL0337WOSEQ_ST25.txt, created on Apr. 29, 2019,
which is 40 KB in size. The information in the electronic format of
the sequence listing is incorporated herein by reference in its
entirety.
FIELD
[0002] Provided herein are compounds, methods, and pharmaceutical
compositions for reducing an amount of Factor XI (FXI) RNA in a
cell or animal, and in certain instances reducing the amount of FXI
protein in a cell or animal. Such compounds, methods, and
pharmaceutical compositions are useful to treat or prevent a
thromboembolic condition. In certain embodiments, the compounds,
methods, and pharmaceutical compositions are useful to treat or
prevent a thromboembolic condition without increasing bleeding
risk. Such thromboembolic conditions include deep vein thrombosis,
venous or arterial thrombosis, pulmonary embolism, myocardial
infarction, stroke, thrombosis associated with chronic kidney
disease or end-stage renal disease (ESRD), including thrombosis
associated with dialysis, or other procoagulant condition.
BACKGROUND
[0003] The circulatory system requires mechanisms that prevent
blood loss, as well as those that counteract inappropriate
intravascular obstructions. Generally, coagulation comprises a
cascade of reactions culminating in the conversion of soluble
fibrinogen to an insoluble fibrin gel. The steps of the cascade
involve the conversion of an inactive zymogen to an activated
enzyme. The active enzyme then catalyzes the next step in the
cascade.
Coagulation Cascade
[0004] The coagulation cascade may be initiated through two
branches, the tissue factor pathway (also "extrinsic pathway"),
which is the primary pathway, and the contact activation pathway
(also "intrinsic pathway").
[0005] The tissue factor pathway is initiated by the cell surface
receptor tissue factor (TF, also referred to as factor III), which
is expressed constitutively by extravascular cells (pericytes,
cardiomyocytes, smooth muscle cells, and keratinocytes) and
expressed by vascular monocytes and endothelial cells upon
induction by inflammatory cytokines or endotoxin. (Drake et al., Am
J Pathol 1989, 134:1087-1097). TF is the high affinity cellular
receptor for coagulation factor VIIa, a serine protease. In the
absence of TF, VIIa has very low catalytic activity, and binding to
TF is necessary to render VIIa functional through an allosteric
mechanism. (Drake et al., Am J Pathol 1989, 134:1087-1097). The
TF-VIIa complex activates factor X to Xa. Xa in turn associates
with its co-factor factor Va into a prothrombinase complex which in
turn activates prothrombin, (also known as factor II or factor 2)
to thrombin (also known as factor IIa, or factor 2a). Thrombin
activates platelets, converts fibrinogen to fibrin and promotes
fibrin cross-linking by activating factor XIII, thus forming a
stable plug at sites where TF is exposed on extravascular cells. In
addition, thrombin reinforces the coagulation cascade response by
activating factors V and VIII.
[0006] The contact activation pathway is triggered by activation of
factor XII to XIIa. Factor XIIa converts XI to XIa, and XIa
converts IX to IXa. IXa associates with its cofactor Villa to
convert X to Xa. The two pathways converge at this point as factor
Xa associates with factor Va to activate prothrombin (factor II) to
thrombin (factor IIa). Factor XI enhances both the formation and
stability of clots in vitro, but is not thought to be involved in
the initiation of clotting. Rather, Factor XI is important in the
propagation phase of clot growth (von de Borne, et al., Blood
Coagulation and Fibrinolysis, 2006, 17:251-257). Additionally,
Factor XI-dependent amplification of thrombin formation leads to
activation of TAFI (thrombin activatable fibrinolysis inhibitor),
which renders clots less sensitive to fibrinolysis (Bouma et al, J
Thromb Haemost 1999; 82: 1703-1708).
Inhibition of Coagulation.
[0007] At least three mechanisms keep the coagulation cascade in
check, namely the action of activated protein C, antithrombin, and
tissue factor pathway inhibitor. Activated protein C is a serine
protease that degrades cofactors Va and VIIIa. Protein C is
activated by thrombin with thrombomodulin, and requires coenzyme
Protein S to function. Antithrombin is a serine protease inhibitor
(serpin) that inhibits serine proteases: thrombin, Xa, XIIa, XIa
and IXa. Tissue factor pathway inhibitor inhibits the action of Xa
and the TF-VIIa complex. (Schwartz A L et al., Trends Cardiovasc
Med. 1997; 7:234-239.)
Disease
[0008] Thrombosis is the pathological development of blood clots,
and an embolism occurs when a blood clot migrates to another part
of the body and interferes with organ function. Thromboembolism may
cause conditions such as deep vein thrombosis, pulmonary embolism,
myocardial infarction, and stroke. Significantly, thromboembolism
is a major cause of morbidity affecting over 2 million Americans
every year. (Adcock et al. American Journal of Clinical Pathology.
1997; 108:434-49). While most cases of thrombosis are due to
acquired extrinsic problems, for example, surgery, cancer,
immobility, some cases are due to a genetic predisposition, for
example, antiphospholipid syndrome and the autosomal dominant
condition, Factor V Leiden. (Bertina R M et al. Nature 1994;
369:64-67.)
Treatment
[0009] The most commonly used anticoagulants, warfarin, heparin,
low molecular weight heparin (LMWH), and newer direct oral
anticoagulants (DOAC), all possess significant drawbacks.
[0010] Warfarin is typically used to treat patients suffering from
atrial fibrillation. The drug interacts with vitamin K-dependent
coagulation factors which include factors II, VII, IX and X.
Anticoagulant proteins C and S are also inhibited by warfarin. Drug
therapy using warfarin is further complicated by the fact that
warfarin interacts with other medications, including drugs used to
treat atrial fibrillation, such as amiodarone. Because therapy with
warfarin is difficult to predict, patients must be carefully
monitored in order to detect any signs of anomalous bleeding.
[0011] Heparin functions by activating antithrombin which inhibits
both thrombin and factor X. (Bjork I, Lindahl U. Mol Cell Biochem.
1982 48: 161-182.) Treatment with heparin may cause an
immunological reaction that makes platelets aggregate within blood
vessels that can lead to thrombosis. This side effect is known as
heparin-induced thrombocytopenia (HIT) resulting in increased
bleeding and requires patient monitoring. Prolonged treatment with
heparin may also lead to osteoporosis. LMWH can also inhibit Factor
II, but to a lesser degree than unfractioned heparin (UFH). LMWH
has been implicated in the development of HIT.
[0012] Several direct oral anticoagulants have been FDA-approved
for the treatment of thrombotic disease, including four Factor Xa
inhibitors Betrixaban, Apixaban, Rivaroxaban, and Edoxaban and one
direct thrombin inhibitor, Dabigatran. (Smith, M., Surg Clin N Am
2018 98:219-238). Rivaroxaban, Dabigatran, and Edoxaban all exhibit
increased bleeding, especially increased GI bleeding risk compared
to warfarin.
[0013] Currently there remains a need for therapies to treat
thromboembolic conditions without risk of increased bleeding. It is
therefore an object herein to provide compounds, methods, and
pharmaceutical compositions for the treatment of such diseases.
BRIEF DESCRIPTION OF THE DRAWINGS
[0014] FIG. 1 shows pharmacodynamic results over time for single
dose cohorts receiving Compound No. 957943, as measured by relative
plasma FXI protein activity. FIG. 1A shows levels of plasma FXI
protein activity. FIG. 1B shows percent change in plasma FXI
protein activity relative to baseline.
[0015] FIG. 2 shows pharmacodynamic results over time for single
dose cohorts receiving Compound No. 957943, as measured by ELISA.
FIG. 2A shows concentrations of plasma FXI protein. FIG. 2B shows
percent change in plasma FXI protein concentrations relative to
baseline.
[0016] FIG. 3 shows pharmacodynamic results over time for multiple
dose cohorts receiving Compound No. 957943, as measured by relative
plasma FXI protein activity. FIG. 3A shows levels of plasma FXI
protein activity. FIG. 3B shows percent change in plasma FXI
protein activity relative to baseline.
[0017] FIG. 4 shows pharmacodynamic results over time for multiple
dose cohorts receiving Compound No. 957943, as measured by ELISA.
FIG. 4A shows concentrations of plasma FXI protein. FIG. 4B shows
change in plasma FXI protein concentrations relative to
baseline.
[0018] FIG. 5 shows pharmacodynamic results over time for multiple
dose cohorts receiving 80 mg Compound No. 957943 every four weeks
for thirteen weeks, as measured by relative plasma FXI protein
activity. FIG. 5A shows levels of plasma FXI protein activity. FIG.
5B shows mean percent change in plasma FXI protein activity.
[0019] FIG. 6 shows pharmacodynamic results over time for multiple
dose cohorts receiving 80 mg Compound No. 957943 every four weeks
for thirteen weeks, as measured by ELISA. FIG. 6A shows plasma FXI
protein concentrations. FIG. 6B shows percent change in plasma FXI
protein concentrations.
SUMMARY OF THE INVENTION
[0020] Provided herein are compounds, methods and pharmaceutical
compositions for reducing an amount of FXI RNA, and in certain
embodiments reducing the amount of FXI protein in a cell or animal.
In certain embodiments, methods and pharmaceutical compositions
disclosed herein reduce FXI protein activity in the blood of an
animal. In certain embodiments, the animal has or is at risk for a
thromboembolic condition. In certain embodiments, the animal has or
is at risk for deep vein thrombosis, venous or arterial thrombosis,
pulmonary embolism, myocardial infarction, stroke, thrombosis
associated with chronic kidney disease or end-stage renal disease
(ESRD), including thrombosis associated with dialysis, or other
procoagulant condition.
[0021] In certain embodiments, compounds useful for reducing a FXI
RNA are oligomeric compounds. In certain embodiments, compounds
useful for reducing a FXI RNA are modified oligonucleotides. In
certain embodiments, compounds useful for reducing a FXI RNA are
oligomeric compounds comprising a conjugate group and a modified
oligonucleotide. In certain embodiments, compounds useful for
reducing a FXI RNA are oligomeric compounds consisting of a
conjugate group and a modified oligonucleotide.
[0022] Also provided are methods useful for treating, preventing,
or ameliorating a thromboembolic condition. In certain embodiments,
the thromboembolic condition is deep vein thrombosis, venous or
arterial thrombosis, pulmonary embolism, myocardial infarction,
stroke, thrombosis associated with chronic kidney disease or
end-stage renal disease (ESRD), including thrombosis associated
with dialysis, or other procoagulant condition.
[0023] Also provided are methods useful treating, preventing, or
ameliorating a thromboembolic condition without increasing bleeding
risk in an individual. In certain embodiments, the individual is at
risk for a thromboembolic condition, including, but not limited to
infarct, thrombosis, embolism, thromboembolism such as deep vein
thrombosis, pulmonary embolism, myocardial infarction, and stroke.
Such diseases, disorders, and conditions can have one or more risk
factors, causes, or outcomes in common. Certain risk factors and
causes for development of a thromboembolic condition include
immobility, surgery (particularly orthopedic surgery), dialysis,
malignancy, pregnancy, older age, use of oral contraceptives,
atrial fibrillation, previous thromboembolic condition, chronic
inflammatory disease, inherited or acquired prothrombotic clotting
disorders and thrombosis associated with chronic kidney disease or
end-stage renal disease (ESRD). Certain outcomes associated with
development of a thromboembolic condition include decreased blood
flow through an affected vessel, death of tissue, and death.
DETAILED DESCRIPTION OF THE INVENTION
[0024] It is to be understood that both the foregoing general
description and the following detailed description are exemplary
and explanatory only and are not restrictive. Herein, the use of
the singular includes the plural unless specifically stated
otherwise. As used herein, the use of "or" means "and/or" unless
stated otherwise. Furthermore, the use of the term "including" as
well as other forms, such as "includes" and "included", is not
limiting. Also, terms such as "element" or "component" encompass
both elements and components comprising one unit and elements and
components that comprise more than one subunit, unless specifically
stated otherwise.
[0025] The section headings used herein are for organizational
purposes only and are not to be construed as limiting the subject
matter described. All documents, or portions of documents, cited in
this application, including, but not limited to, patents, patent
applications, articles, books, and treatises, are hereby expressly
incorporated-by-reference for the portions of the document
discussed herein, as well as in their entirety.
Definitions
[0026] Unless specific definitions are provided, the nomenclature
used in connection with, and the procedures and techniques of,
analytical chemistry, synthetic organic chemistry, and medicinal
and pharmaceutical chemistry described herein are those well known
and commonly used in the art. Where permitted, all patents,
applications, published applications and other publications and
other data referred to throughout in the disclosure are
incorporated by reference herein in their entirety.
[0027] Unless otherwise indicated, the following terms have the
following meanings:
Definitions
[0028] As used herein, "2'-deoxynucleoside" means a nucleoside
comprising a 2'-H(H) deoxyribosyl sugar moiety, as found in
naturally occurring deoxyribonucleic acids (DNA). In certain
embodiments, a 2'-deoxynucleoside may comprise a modified
nucleobase or may comprise an RNA nucleobase (uracil).
[0029] As used herein, "2'-substituted nucleoside" means a
nucleoside comprising a 2'-substituted sugar moiety. As used
herein, "2'-substituted" in reference to a sugar moiety means a
sugar moiety comprising at least one 2'-substituent group other
than H or OH.
[0030] As used herein, "5-methyl cytosine" means a cytosine
modified with a methyl group attached to the 5 position. A 5-methyl
cytosine is a modified nucleobase.
[0031] As used herein, "about" means plus or minus 7% of the
provided value.
[0032] As used herein, "administering" means providing a
pharmaceutical agent to an animal.
[0033] As used herein, "animal" means a human or non-human animal.
In certain embodiments, the animal is a human.
[0034] As used herein, "antisense activity" means any detectable
and/or measurable change attributable to the hybridization of an
antisense compound to its target nucleic acid. In certain
embodiments, antisense activity is a decrease in the amount or
expression of a target nucleic acid or protein encoded by such
target nucleic acid compared to target nucleic acid levels or
target protein levels in the absence of the antisense compound.
[0035] As used herein, "antisense compound" means an oligomeric
compound capable of achieving at least one antisense activity.
[0036] As used herein, "cleavable moiety" means a bond or group of
atoms that is cleaved under physiological conditions, for example,
inside a cell or an animal.
[0037] As used herein, "complementary" in reference to an
oligonucleotide means that at least 70% of the nucleobases of the
oligonucleotide or one or more regions thereof and the nucleobases
of another nucleic acid or one or more regions thereof are capable
of hydrogen bonding with one another when the nucleobase sequence
of the oligonucleotide and the other nucleic acid are aligned in
opposing directions. Complementary nucleobases means nucleobases
that are capable of forming hydrogen bonds with one another.
Complementary nucleobase pairs include adenine (A) and thymine (T),
adenine (A) and uracil (U), cytosine (C) and guanine (G), 5-methyl
cytosine (mC) and guanine (G). Complementary oligonucleotides
and/or nucleic acids need not have nucleobase complementarity at
each nucleoside. Rather, some mismatches are tolerated. As used
herein, "fully complementary" or "100% complementary" in reference
to oligonucleotides means that oligonucleotides are complementary
to another oligonucleotide or nucleic acid at each nucleoside of
the oligonucleotide.
[0038] As used herein, "conjugate group" means a group of atoms
that is directly attached to an oligonucleotide. Conjugate groups
include a conjugate moiety and a conjugate linker that attaches the
conjugate moiety to the oligonucleotide.
[0039] As used herein, "conjugate linker" means a single bond or
group of atoms comprising at least one bond that connects a
conjugate moiety to an oligonucleotide. In certain embodiments, a
conjugate linker comprises a cleavable moiety.
[0040] As used herein, "conjugate moiety" means a group of atoms
that is attached to an oligonucleotide via a conjugate linker. In
certain embodiments, a conjugate moiety comprises a cell-targeting
moiety.
[0041] As used herein, "contiguous" in the context of an
oligonucleotide refers to nucleosides, nucleobases, sugar moieties,
or internucleoside linkages that are immediately adjacent to each
other. For example, "contiguous nucleobases" means nucleobases that
are immediately adjacent to each other in a sequence.
[0042] As used herein, "chirally enriched population" means a
plurality of molecules of identical molecular formula, wherein the
number or percentage of molecules within the population that
contain a particular stereochemical configuration at a particular
chiral center is greater than the number or percentage of molecules
expected to contain the same particular stereochemical
configuration at the same particular chiral center within the
population if the particular chiral center were stereorandom.
Chirally enriched populations of molecules having multiple chiral
centers within each molecule may contain one or more stereorandom
chiral centers. In certain embodiments, the molecules are
oligomeric compounds disclosed herein. In certain embodiments, the
oligomeric compounds are antisense compounds. In certain
embodiments, the molecules are modified oligonucleotides. In
certain embodiments, the molecules are oligomeric compounds
comprising modified oligonucleotides.
[0043] As used herein, "dosage unit" means a formulation of an
oligomeric compound, or pharmaceutical composition thereof, for
administration, wherein the oligomeric compound is provided at a
quantity of a single selected dose. The term dosage unit, as used
herein, may comprise packaging or a container that contains the
pharmaceutical composition, such as a vial or syringe. As used
herein, "gapmer" means a modified oligonucleotide comprising an
internal region having a plurality of nucleosides that support
RNase H cleavage positioned between external regions having one or
more nucleosides, wherein the nucleosides comprising the internal
region are chemically distinct from the nucleoside or nucleosides
comprising the external regions. The internal region may be
referred to as the "gap" and the external regions may be referred
to as the "wings." Unless otherwise indicated, "gapmer" refers to a
sugar motif. Unless otherwise indicated, the sugar moieties of the
nucleosides of the gap of a gapmer are unmodified 2'-deoxyribosyl.
Thus, the term "MOE gapmer" indicates a gapmer having a sugar motif
of 2'-MOE nucleosides in both wings and a gap of
2'-deoxynucleosides. Unless otherwise indicated, a MOE gapmer may
comprise one or more modified internucleoside linkages and/or
modified nucleobases and such modifications do not necessarily
follow the gapmer pattern of the sugar modifications.
[0044] As used herein, "hybridization" means the pairing or
annealing of complementary oligonucleotides and/or nucleic acids.
While not limited to a particular mechanism, the most common
mechanism of hybridization involves hydrogen bonding, which may be
Watson-Crick, Hoogsteen or reversed Hoogsteen hydrogen bonding,
between complementary nucleobases.
[0045] As used herein, "identifying an animal at risk for
developing a thromboembolic condition" means identifying an animal
having been diagnosed with a thromboembolic condition or
identifying an animal predisposed to develop a thromboembolic
condition. Individuals predisposed to develop a thromboembolic
condition include those having one or more risk factors for
thromboembolic conditions including immobility, surgery
(particularly orthopedic surgery), dialysis, malignancy, pregnancy,
older age, use of oral contraceptives, inherited or acquired
prothrombotic clotting disorders, chronic kidney disease, and
end-stage renal disease (ESRD). Such identification may be
accomplished by any method including evaluating an individual's
medical history and standard clinical tests or assessments.
[0046] As used herein, the term "internucleoside linkage" is the
covalent linkage between adjacent nucleosides in an
oligonucleotide. As used herein "modified internucleoside linkage"
means any internucleoside linkage other than a phosphodiester
internucleoside linkage. "Phosphorothioate internucleoside linkage"
is a modified internucleoside linkage in which one of the
non-bridging oxygen atoms of a phosphodiester internucleoside
linkage is replaced with a sulfur atom.
[0047] As used herein, the term "linker region" in reference to a
conjugate moiety refers that part of a conjugate linker that is not
a cleavable moiety.
[0048] As used herein, "non-bicyclic modified sugar moiety" means a
modified sugar moiety that comprises a modification, such as a
substituent, that does not form a bridge between two atoms of the
sugar to form a second ring.
[0049] As used herein, "mismatch" or "non-complementary" means a
nucleobase of a first oligonucleotide that is not complementary
with the corresponding nucleobase of a second oligonucleotide or
target nucleic acid when the first and second oligonucleotide are
aligned.
[0050] As used herein, "MOE" means methoxyethyl. "2'-MOE" or
"2'-MOE modified sugar" means a 2'-OCH.sub.2CH.sub.2OCH.sub.3 group
in place of the 2'-OH group of a ribosyl sugar moiety.
[0051] As used herein, "2'-MOE nucleoside" means a nucleoside
comprising a 2'-MOE modified sugar. As used herein, "monthly" means
every 28 to 31 days.
[0052] As used herein, "motif" means the pattern of unmodified
and/or modified sugar moieties, nucleobases, and/or internucleoside
linkages, in an oligonucleotide.
[0053] As used herein, "nucleobase" means an unmodified nucleobase
or a modified nucleobase. As used herein an "unmodified nucleobase"
is adenine (A), thymine (T), cytosine (C), uracil (U), and guanine
(G). As used herein, a "modified nucleobase" is a group of atoms
other than unmodified A, T, C, U, or G capable of pairing with at
least one unmodified nucleobase. A "5-methyl cytosine" is a
modified nucleobase. A universal base is a modified nucleobase that
can pair with any one of the five unmodified nucleobases. As used
herein, "nucleobase sequence" means the order of contiguous
nucleobases in a nucleic acid or oligonucleotide independent of any
sugar or internucleoside linkage modification.
[0054] As used herein, "nucleoside" means a compound comprising a
nucleobase and a sugar moiety. The nucleobase and sugar moiety are
each, independently, unmodified or modified. As used herein,
"modified nucleoside" means a nucleoside comprising a modified
nucleobase and/or a modified sugar moiety. Modified nucleosides
include abasic nucleosides, which lack a nucleobase. "Linked
nucleosides" are nucleosides that are connected in a contiguous
sequence (i.e., no additional nucleosides are presented between
those that are linked).
[0055] As used herein, "oligomeric compound" means an
oligonucleotide and optionally one or more additional features,
such as a conjugate group or terminal group. An oligomeric compound
may be paired with a second oligomeric compound that is
complementary to the first oligomeric compound or may be unpaired.
A "singled-stranded oligomeric compound" is an unpaired oligomeric
compound. The term "oligomeric duplex" means a duplex formed by two
oligomeric compounds having complementary nucleobase sequences.
Each oligomeric compound of an oligomeric duplex may be referred to
as a "duplexed oligomeric compound."
[0056] As used herein, "oligonucleotide" means a strand of linked
nucleosides connected via internucleoside linkages, wherein each
nucleoside and internucleoside linkage may be modified or
unmodified. Unless otherwise indicated, oligonucleotides consist of
8-50 linked nucleosides. As used herein, "modified oligonucleotide"
means an oligonucleotide, wherein at least one nucleoside or
internucleoside linkage is modified. As used herein, "unmodified
oligonucleotide" means an oligonucleotide that does not comprise
any nucleoside modifications or internucleoside modifications.
[0057] As used herein, "pharmaceutically acceptable carrier or
diluent" means any substance suitable for use in administering to
an animal. Certain such carriers enable pharmaceutical compositions
to be formulated as, for example, tablets, pills, dragees,
capsules, liquids, gels, syrups, slurries, suspension and lozenges
for the oral ingestion by an animal. In certain embodiments, a
pharmaceutically acceptable carrier or diluent is sterile water,
distilled water for injection, sterile saline, sterile buffer
solution or sterile artificial cerebrospinal fluid.
[0058] As used herein "pharmaceutically acceptable salts" means
physiologically and pharmaceutically acceptable salts of compounds.
Pharmaceutically acceptable salts retain the desired biological
activity of the parent compound and do not impart undesired
toxicological effects thereto.
[0059] As used herein "pharmaceutical composition" means a mixture
of substances suitable for administering to an animal. For example,
a pharmaceutical composition may comprise an oligomeric compound
and a sterile aqueous solution. In certain embodiments, a
pharmaceutical composition shows activity in free uptake assay in
certain cell lines.
[0060] As used herein, "reducing or inhibiting the amount or
activity" refers to a reduction or blockade of the transcriptional
expression or activity relative to the transcriptional expression
or activity in an untreated or control sample and does not
necessarily indicate a total elimination of transcriptional
expression or activity.
[0061] As used herein, "RNAi compound" means an antisense compound
that acts, at least in part, through RISC or Ago2 to modulate a
target nucleic acid and/or protein encoded by a target nucleic
acid. RNAi compounds include, but are not limited to
double-stranded siRNA, single-stranded RNA (ssRNA), and microRNA,
including microRNA mimics. In certain embodiments, an RNAi compound
modulates the amount, activity, and/or splicing of a target nucleic
acid. The term RNAi compound excludes antisense compounds that act
through RNase H.
[0062] As used herein, "self-complementary" in reference to an
oligonucleotide means an oligonucleotide that at least partially
hybridizes to itself.
[0063] As used herein, "stereorandom chiral center" in the context
of a population of molecules of identical molecular formula means a
chiral center having a random stereochemical configuration. For
example, in a population of molecules comprising a stereorandom
chiral center, the number of molecules having the (S) configuration
of the stereorandom chiral center may be but is not necessarily the
same as the number of molecules having the (R) configuration of the
stereorandom chiral center. The stereochemical configuration of a
chiral center is considered random when it is the results of a
synthetic method that is not designed to control the stereochemical
configuration. In certain embodiments, a stereorandom chiral center
is a stereorandom phosphorothioate internucleoside linkage.
[0064] As used herein, "sugar moiety" means an unmodified sugar
moiety or a modified sugar moiety. As used herein, "unmodified
sugar moiety" means a 2'-OH(H) ribosyl moiety, as found in RNA (an
"unmodified RNA sugar moiety"), or a 2'-H(H) deoxyribosyl moiety,
as found in DNA (an "unmodified DNA sugar moiety"). Unmodified
sugar moieties have one hydrogen at each of the 1', 3', and 4'
positions, an oxygen at the 3' position, and two hydrogens at the
5' position. As used herein, "modified sugar moiety" or "modified
sugar" means a modified furanosyl sugar moiety or a sugar
surrogate.
[0065] As used herein, "sugar surrogate" means a modified sugar
moiety having other than a furanosyl moiety that can link a
nucleobase to another group, such as an internucleoside linkage,
conjugate group, or terminal group in an oligonucleotide. Modified
nucleosides comprising sugar surrogates can be incorporated into
one or more positions within an oligonucleotide and such
oligonucleotides are capable of hybridizing to complementary
oligomeric compounds or target nucleic acids.
[0066] As used herein, "thromboembolic condition" means any disease
or condition involving an embolism caused by a thrombus. Examples
of such diseases=and conditions include the categories of
thrombosis, embolism, and thromboembolism. In certain embodiments,
such diseases and conditions include deep vein thrombosis, venous
or arterial thrombosis, pulmonary embolism, myocardial infarction,
stroke, thrombosis associated with chronic kidney disease or
end-stage renal disease (ESRD), including thrombosis associated
with dialysis, or other procoagulant condition. Thromboembolic
conditions may also be referred to as thromboembolic events or
thrombotic events.
[0067] As used herein, "target nucleic acid" and "target RNA" mean
a nucleic acid that an antisense compound is designed to
affect.
[0068] As used herein, "target region" means a portion of a target
nucleic acid to which an oligomeric compound is designed to
hybridize.
[0069] As used herein, "therapeutically effective amount" means an
amount of a pharmaceutical agent that provides a therapeutic
benefit to an animal. For example, a therapeutically effective
amount of a pharmaceutical agent treats, prevents, or ameliorates a
thromboembolic condition.
[0070] As used herein, "weekly" means every six to eight days.
CERTAIN EMBODIMENTS
[0071] The present disclosure provides the following non-limiting
numbered embodiments:
[0072] Embodiment 1. An oligomeric compound according to the
following formula:
##STR00001##
or a salt thereof.
[0073] Embodiment 2. An oligomeric compound according to the
following formula:
##STR00002##
[0074] Embodiment 3. An oligomeric compound comprising a modified
oligonucleotide according to the following formula:
##STR00003## [0075] wherein one of R.sub.1 or R.sub.2 is a
conjugate group comprising at least one, two, or three GalNAc
ligands, and the other of R.sub.1 or R.sub.2 is --OH; [0076] or a
salt thereof.
[0077] Embodiment 4. An oligomeric compound comprising a modified
oligonucleotide according to the following formula:
(THA-GalNAc.sub.3)o Aes mCeo Geo Geo mCeo Ads Tds Tds Gds Gds Tds
Gds mCds Ads mCds Aeo Geo Tes Tes Te (SEQ ID NO: 3); wherein,
(THA-GalNAc3)o is represented by the following structure:
##STR00004##
and wherein,
[0078] A=an adenine nucleobase,
[0079] mC=a 5'-methyl cytosine nucleobase,
[0080] G=a guanine nucleobase,
[0081] T=a thymine nucleobase,
[0082] e=a 2'-MOE modified sugar,
[0083] d=a 2'-deoxyribose sugar,
[0084] s=a phosphorothioate internucleoside linkage, and
[0085] o=a phosphodiester internucleoside linkage;
[0086] or a salt thereof.
[0087] Embodiment 5. The oligomeric compound of any one of
embodiments 1, 3 or 5, which is a sodium salt.
[0088] Embodiment 6. A chirally enriched population of the
oligomeric compound of any of embodiments 1-5 wherein the
population is enriched for oligomeric compounds having a modified
oligonucleotide comprising at least one particular phosphorothioate
internucleoside linkage having a particular stereochemical
configuration.
[0089] Embodiment 7. The chirally enriched population of embodiment
6, wherein the population is enriched for oligomeric compounds
having a modified oligonucleotide comprising at least one
particular phosphorothioate internucleoside linkage having the (Sp)
configuration.
[0090] Embodiment 8. The chirally enriched population of embodiment
6 or 7, wherein the population is enriched for oligomeric compounds
having a modified oligonucleotides comprising at least one
particular phosphorothioate internucleoside linkage having the (Rp)
configuration.
[0091] Embodiment 9. The chirally enriched population of embodiment
6, wherein the population is enriched for oligomeric compounds
having a modified oligonucleotide having a particular,
independently selected stereochemical configuration at each
phosphorothioate internucleoside linkage.
[0092] Embodiment 10. The chirally enriched population of
embodiment 9, wherein the population is enriched for oligomeric
compounds having a modified oligonucleotide having the (Sp)
configuration at each phosphorothioate internucleoside linkage.
[0093] Embodiment 11. The chirally enriched population of
embodiment 9, wherein the population is enriched for oligomeric
compounds having a modified oligonucleotide having the (Rp)
configuration at each phosphorothioate internucleoside linkage.
[0094] Embodiment 12. The chirally enriched population of
embodiment 6 or 9 wherein the population is enriched for oligomeric
compounds having a modified oligonucleotide having at least 3
contiguous phosphorothioate internucleoside linkages in the
Sp-Sp-Rp configurations, in the 5' to 3' direction.
[0095] Embodiment 13. A population of oligomeric compounds having a
modified oligonucleotide of any of embodiments 1-5, wherein all of
the phosphorothioate internucleoside linkages of the modified
oligonucleotide are stereorandom.
[0096] Embodiment 14. A pharmaceutical composition comprising the
oligomeric compound of any one of embodiments 1-5, the chirally
enriched population of any one of embodiments 6-12, or the
population of embodiment 13, and a pharmaceutically acceptable
carrier or diluent.
[0097] Embodiment 15. The pharmaceutical composition of embodiment
14, wherein the pharmaceutically acceptable diluent is phosphate
buffered saline.
[0098] Embodiment 16. The pharmaceutical composition of embodiment
14, wherein the pharmaceutical composition consists or consists
essentially of the oligomeric compound and phosphate buffered
saline.
[0099] Embodiment 17. The pharmaceutical composition of any one of
embodiments 14-16, wherein the concentration of the oligomeric
compound in the pharmaceutically acceptable carrier or diluent is
selected from: [0100] a) 5 mg/ml, 10 mg/ml, 15 mg/ml, 20 mg/ml, 25
mg/ml, 30 mg/ml, 35 mg/ml, 40 mg/ml, 45 mg/ml, 50 mg/ml, 55 mg/ml,
60 mg/ml, 65 mg/ml, 70 mg/ml, 75 mg/ml, 80 mg/ml, 85 mg/ml, 90
mg/ml, 95 mg/ml, 100 mg/ml, 105 mg/ml, 110 mg/ml, 115 mg/ml, 120
mg/ml, 125 mg/ml, 130 mg/ml, 135 mg/ml, 140 mg/ml, 145 mg/ml, 150
mg/ml, 155 mg/ml, 160 mg/ml; [0101] b) about 5 mg/ml, about 10
mg/ml, about 15 mg/ml, about 20 mg/ml, about 25 mg/ml, about 30
mg/ml, about 35 mg/ml, about 40 mg/ml, about 45 mg/ml, about 50
mg/ml, about 55 mg/ml, about 60 mg/ml, about 65 mg/ml, about 70
mg/ml, about 75 mg/ml, about 80 mg/ml, about 85 mg/ml, about 90
mg/ml, about 95 mg/ml, about 100 mg/ml, about 105 mg/ml, about 110
mg/ml, about 115 mg/ml, about 120 mg/ml, about 125 mg/ml, about 130
mg/ml, about 135 mg/ml, about 140 mg/ml, about 145 mg/ml, about 150
mg/ml, about 155 mg/ml, about 160 mg/ml; and [0102] c) 20 mg/ml, 21
mg/ml, 22 mg/ml, 23 mg/ml, 24 mg/ml, 25 mg/ml, 26 mg/ml, 27 mg/ml,
28 mg/ml, 29 mg/ml, 30 mg/ml, 31 mg/ml, 32 mg/ml, 33 mg/ml, 34
mg/ml, 35 mg/ml, 36 mg/ml, 37 mg/ml, 38 mg/ml, 39 mg/ml, 40 mg/ml,
41 mg/ml, 42 mg/ml, 43 mg/ml, 44 mg/ml, 45 mg/ml, 46 mg/ml, 47
mg/ml, 48 mg/ml, 49 mg/ml, 50 mg/ml, 51 mg/ml, 52 mg/ml, 53 mg/ml,
54 mg/ml, 55 mg/ml, 56 mg/ml, 57 mg/ml, 58 mg/ml, 59 mg/ml, 60
mg/ml, 61 mg/ml, 62 mg/ml, 63 mg/ml, 64 mg/ml, 65 mg/ml, 66 mg/ml,
67 mg/ml, 68 mg/ml, 69 mg/ml, 70 mg/ml, 71 mg/ml, 72 mg/ml, 73
mg/ml, 74 mg/ml, 75 mg/ml, 76 mg/ml, 77 mg/ml, 78 mg/ml, 79 mg/ml,
80 mg/ml, 81 mg/ml, 82 mg/ml, 83 mg/ml, 84 mg/ml, 85 mg/ml, 86
mg/ml, 87 mg/ml, 88 mg/ml, 89 mg/ml, 90 mg/ml, 91 mg/ml, 92 mg/ml,
93 mg/ml, 94 mg/ml, 95 mg/ml, 96 mg/ml, 97 mg/ml, 98 mg/ml, 99
mg/ml, 100 mg/ml, 101 mg/ml, 102 mg/ml, 103 mg/ml, 104 mg/ml, 105
mg/ml, 106 mg/ml, 107 mg/ml, 108 mg/ml, 109 mg/ml, 110 mg/ml, 111
mg/ml, 112 mg/ml, 113 mg/ml, 114 mg/ml, 115 mg/ml, 116 mg/ml, 117
mg/ml, 118 mg/ml, 119 mg/ml, 120 mg/ml, 121 mg/ml, 122 mg/ml, 123
mg/ml, 124 mg/ml, 125 mg/ml, 126 mg/ml, 127 mg/ml, 128 mg/ml, 129
mg/ml, 130 mg/ml, 131 mg/ml, 132 mg/ml, 133 mg/ml, 134 mg/ml, 135
mg/ml, 136 mg/ml, 137 mg/ml, 138 mg/ml, 139 mg/ml, 140 mg/ml;
[0103] d) about 20 mg/ml, about 21 mg/ml, about 22 mg/ml, about 23
mg/ml, about 24 mg/ml, about 25 mg/ml, about 26 mg/ml, about 27
mg/ml, about 28 mg/ml, about 29 mg/ml, about 30 mg/ml, about 31
mg/ml, about 32 mg/ml, about 33 mg/ml, about 34 mg/ml, about 35
mg/ml, about 36 mg/ml, about 37 mg/ml, about 38 mg/ml, about 39
mg/ml, about 40 mg/ml, about 41 mg/ml, about 42 mg/ml, about 43
mg/ml, about 44 mg/ml, about 45 mg/ml, about 46 mg/ml, about 47
mg/ml, about 48 mg/ml, about 49 mg/ml, about 50 mg/ml, about 51
mg/ml, about 52 mg/ml, about 53 mg/ml, about 54 mg/ml, about 55
mg/ml, about 56 mg/ml, about 57 mg/ml, about 58 mg/ml, about 59
mg/ml, about 60 mg/ml, about 61 mg/ml, about 62 mg/ml, about 63
mg/ml, about 64 mg/ml, about 65 mg/ml, about 66 mg/ml, about 67
mg/ml, about 68 mg/ml, about 69 mg/ml, about 70 mg/ml, about 71
mg/ml, about 72 mg/ml, about 73 mg/ml, about 74 mg/ml, about 75
mg/ml, about 76 mg/ml, about 77 mg/ml, about 78 mg/ml, about 79
mg/ml, about 80 mg/ml, about 81 mg/ml, about 82 mg/ml, about 83
mg/ml, about 84 mg/ml, about 85 mg/ml, about 86 mg/ml, about 87
mg/ml, about 88 mg/ml, about 89 mg/ml, about 90 mg/ml, about 91
mg/ml, about 92 mg/ml, about 93 mg/ml, about 94 mg/ml, about 95
mg/ml, about 96 mg/ml, about 97 mg/ml, about 98 mg/ml, about 99
mg/ml, about 100 mg/ml, about 101 mg/ml, about 102 mg/ml, about 103
mg/ml, about 104 mg/ml, about 105 mg/ml, about 106 mg/ml, about 107
mg/ml, about 108 mg/ml, about 109 mg/ml, about 110 mg/ml, about 111
mg/ml, about 112 mg/ml, about 113 mg/ml, about 114 mg/ml, about 115
mg/ml, about 116 mg/ml, about 117 mg/ml, about 118 mg/ml, about 119
mg/ml, about 120 mg/ml, about 121 mg/ml, about 122 mg/ml, about 123
mg/ml, about 124 mg/ml, about 125 mg/ml, about 126 mg/ml, about 127
mg/ml, about 128 mg/ml, about 129 mg/ml, about 130 mg/ml, about 131
mg/ml, about 132 mg/ml, about 133 mg/ml, about 134 mg/ml, about 135
mg/ml, about 136 mg/ml, about 137 mg/ml, about 138 mg/ml, about 139
mg/ml, and about 140 mg/ml.
[0104] Embodiment 18. The pharmaceutical composition of any one of
embodiments 14-16, wherein the concentration of the oligomeric
compound in the pharmaceutically acceptable carrier or diluent is
selected from 20 mg/ml to 180 mg/ml, 20 mg/ml to 170 mg, 20 mg/ml
to 160 mg/ml, 20 mg/ml to 150 mg/ml, 20 mg/ml to 140 mg/ml, 20
mg/ml to 130 mg/ml, 20 mg/ml to 120 mg/ml, 20 mg/ml to 110 mg/ml,
20 mg/ml to 100 mg/ml, 20 mg/ml to 90 mg/ml, 20 mg/ml to 80 mg/ml,
20 mg/ml to 70 mg/ml, 20 mg/ml to 60 mg/ml, 20 mg/ml to 50 mg/ml,
20 mg/ml to 40 mg/ml, 20 mg/ml to 30 mg/ml, 30 mg/ml to 180 mg/ml,
30 mg/ml to 170 mg, 30 mg/ml to 160 mg/ml, 30 mg/ml to 150 mg/ml,
30 mg/ml to 140 mg/ml, 30 mg/ml to 130 mg/ml, 30 mg/ml to 120
mg/ml, 30 mg/ml to 110 mg/ml, 30 mg/ml to 100 mg/ml, 30 mg/ml to 90
mg/ml, 30 mg/ml to 80 mg/ml, 30 mg/ml to 70 mg/ml, 30 mg/ml to 60
mg/ml, 30 mg/ml to 50 mg/ml, 30 mg/ml to 40 mg/ml, 40 mg/ml to 180
mg/ml, 40 mg/ml to 170 mg, 40 mg/ml to 160 mg/ml, 40 mg/ml to 150
mg/ml, 40 mg/ml to 140 mg/ml, 40 mg/ml to 130 mg/ml, 40 mg/ml to
120 mg/ml, 40 mg/ml to 110 mg/ml, 40 mg/ml to 100 mg/ml, 40 mg/ml
to 90 mg/ml, 40 mg/ml to 80 mg/ml, 40 mg/ml to 70 mg/ml, 40 mg/ml
to 60 mg/ml, 40 mg/ml to 50 mg/ml, 50 mg/ml to 180 mg/ml, 50 mg/ml
to 170 mg, 50 mg/ml to 160 mg/ml, 50 mg/ml to 150 mg/ml, 50 mg/ml
to 140 mg/ml, 50 mg/ml to 130 mg/ml, 50 mg/ml to 120 mg/ml, 50
mg/ml to 110 mg/ml, 50 mg/ml to 100 mg/ml, 50 mg/ml to 90 mg/ml, 50
mg/ml to 80 mg/ml, 50 mg/ml to 70 mg/ml, 50 mg/ml to 60 mg/ml, 60
mg/ml to 180 mg/ml, 60 mg/ml to 170 mg, 60 mg/ml to 160 mg/ml, 60
mg/ml to 150 mg/ml, 60 mg/ml to 140 mg/ml, 60 mg/ml to 130 mg/ml,
60 mg/ml to 120 mg/ml, 60 mg/ml to 110 mg/ml, 60 mg/ml to 100
mg/ml, 60 mg/ml to 90 mg/ml, 60 mg/ml to 80 mg/ml, 60 mg/ml to 70
mg/ml, 70 mg/ml to 180 mg/ml, 70 mg/ml to 170 mg, 70 mg/ml to 160
mg/ml, 70 mg/ml to 150 mg/ml, 70 mg/ml to 140 mg/ml, 70 mg/ml to
130 mg/ml, 70 mg/ml to 120 mg/ml, 70 mg/ml to 110 mg/ml, 70 mg/ml
to 100 mg/ml, 70 mg/ml to 90 mg/ml, 70 mg/ml to 80 mg/ml, 80 mg/ml
to 180 mg/ml, 80 mg/ml to 170 mg, 80 mg/ml to 160 mg/ml, 80 mg/ml
to 150 mg/ml, 80 mg/ml to 140 mg/ml, 80 mg/ml to 130 mg/ml, 80
mg/ml to 120 mg/ml, 80 mg/ml to 110 mg/ml, 80 mg/ml to 100 mg/ml,
80 mg/ml to 90 mg/ml, 90 mg/ml to 180 mg/ml, 90 mg/ml to 170 mg, 90
mg/ml to 160 mg/ml, 90 mg/ml to 150 mg/ml, 90 mg/ml to 140 mg/ml,
90 mg/ml to 130 mg/ml, 90 mg/ml to 120 mg/ml, 90 mg/ml to 110
mg/ml, 90 mg/ml to 100 mg/ml, 100 mg/ml to 180 mg/ml, 100 mg/ml to
170 mg, 100 mg/ml to 160 mg/ml, 100 mg/ml to 150 mg/ml, 100 mg/ml
to 140 mg/ml, 100 mg/ml to 130 mg/ml, 100 mg/ml to 120 mg/ml, 100
mg/ml to 110 mg/ml, 110 mg/ml to 180 mg/ml, 110 mg/ml to 170 mg,
110 mg/ml to 160 mg/ml, 110 mg/ml to 150 mg/ml, 110 mg/ml to 140
mg/ml, 110 mg/ml to 130 mg/ml, 110 mg/ml to 120 mg/ml, 120 mg/ml to
180 mg/ml, 120 mg/ml to 170 mg, 120 mg/ml to 160 mg/ml, 120 mg/ml
to 150 mg/ml, 120 mg/ml to 140 mg/ml, 120 mg/ml to 130 mg/ml, 130
mg/ml to 180 mg/ml, 130 mg/ml to 170 mg, 130 mg/ml to 160 mg/ml,
130 mg/ml to 150 mg/ml, 130 mg/ml to 140 mg/ml, 140 mg/ml to 180
mg/ml, 140 mg/ml to 170 mg/ml, 140 mg/ml to 160 mg/ml, 140 mg/ml to
150 mg/ml, 150 mg/ml to 180 mg/ml, 150 mg/ml to 170 mg/ml, 150
mg/ml to 160 mg/ml, 160 mg/ml to 180 mg/ml, 160 mg/ml to 170 mg/ml,
and 170 mg/ml to 180 mg/ml.
[0105] Embodiment 19. The pharmaceutical composition of any one of
embodiments 14-18, wherein the pharmaceutical composition is in a
form of a dosage unit.
[0106] Embodiment 20. The pharmaceutical composition of embodiment
19, wherein the oligomeric compound is present in the dosage unit
at an amount selected from: [0107] a) 5 mg, 10 mg, 15 mg, 20 mg, 25
mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg, 65 mg, 70 mg,
75 mg, 80 mg, 85 mg, 90 mg, 95 mg, 100 mg, 105 mg, 110 mg, 115 mg,
120 mg, 125 mg, 130 mg, 135 mg, 140 mg, 145 mg, 150 mg, 155 mg, 160
mg, 165 mg, 170 mg, 175 mg, 180 mg, 185 mg, 190 mg, 195 mg, 200 mg,
205 mg, 210 mg, 215 mg, 220 mg, 225 mg, 230 mg, 235 mg, 240 mg, 245
mg, 250 mg, 255 mg, 260 mg, 265 mg, 270 mg, 275 mg, 280 mg, 285 mg,
290 mg, 295 mg, 300 mg; [0108] b) about 5 mg, about 10 mg, about 15
mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40
mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65
mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, about 90
mg, about 95 mg, about 100 mg, about 105 mg, about 110 mg, about
115 mg, about 120 mg, about 125 mg, about 130 mg, about 135 mg,
about 140 mg, about 145 mg, about 150 mg, about 155 mg, about 160
mg, about 165 mg, about 170 mg, about 175 mg, about 180 mg, about
185 mg, about 190 mg, about 195 mg, about 200 mg, about 205 mg,
about 210 mg, about 215 mg, about 220 mg, about 225 mg, about 230
mg, about 235 mg, about 240 mg, about 245 mg, about 250 mg, about
255 mg, about 260 mg, about 265 mg, about 270 mg, about 275 mg,
about 280 mg, about 285 mg, about 290 mg, about 295 mg, about 300
mg; [0109] c) 20 mg, 21 mg, 22 mg, 23 mg, 24 mg, 25 mg, 26 mg, 27
mg, 28 mg, 29 mg, 30 mg, 31 mg, 32 mg, 33 mg, 34 mg, 35 mg, 36 mg,
37 mg, 38 mg, 39 mg, 40 mg, 41 mg, 42 mg, 43 mg, 44 mg, 45 mg, 46
mg, 47 mg, 48 mg, 49 mg, 50 mg, 51 mg, 52 mg, 53 mg, 54 mg, 55 mg,
56 mg, 57 mg, 58 mg, 59 mg, 60 mg, 61 mg, 62 mg, 63 mg, 64 mg, 65
mg, 66 mg, 67 mg, 68 mg, 69 mg, 70 mg, 71 mg, 72 mg, 73 mg, 74 mg,
75 mg, 76 mg, 77 mg, 78 mg, 79 mg, 80 mg, 81 mg, 82 mg, 83 mg, 84
mg, 85 mg, 86 mg, 87 mg, 88 mg, 89 mg, 90 mg, 91 mg, 92 mg, 93 mg,
94 mg, 95 mg, 96 mg, 97 mg, 98 mg, 99 mg, 100 mg, 101 mg, 102 mg,
103 mg, 104 mg, 105 mg, 106 mg, 107 mg, 108 mg, 109 mg, 110 mg, 111
mg, 112 mg, 113 mg, 114 mg, 115 mg, 116 mg, 117 mg, 118 mg, 119 mg,
120 mg, 121 mg, 122 mg, 123 mg, 124 mg, 125 mg, 126 mg, 127 mg, 128
mg, 129 mg, 130 mg, 131 mg, 132 mg, 133 mg, 134 mg, 135 mg, 136 mg,
137 mg, 138 mg, 139 mg, 140 mg; [0110] d) about 20 mg, about 21 mg,
about 22 mg, about 23 mg, about 24 mg, about 25 mg, about 26 mg,
about 27 mg, about 28 mg, about 29 mg, about 30 mg, about 31 mg,
about 32 mg, about 33 mg, about 34 mg, about 35 mg, about 36 mg,
about 37 mg, about 38 mg, about 39 mg, about 40 mg, about 41 mg,
about 42 mg, about 43 mg, about 44 mg, about 45 mg, about 46 mg,
about 47 mg, about 48 mg, about 49 mg, about 50 mg, about 51 mg,
about 52 mg, about 53 mg, about 54 mg, about 55 mg, about 56 mg,
about 57 mg, about 58 mg, about 59 mg, about 60 mg, about 61 mg,
about 62 mg, about 63 mg, about 64 mg, about 65 mg, about 66 mg,
about 67 mg, about 68 mg, about 69 mg, about 70 mg, about 71 mg,
about 72 mg, about 73 mg, about 74 mg, about 75 mg, about 76 mg,
about 77 mg, about 78 mg, about 79 mg, about 80 mg, about 81 mg,
about 82 mg, about 83 mg, about 84 mg, about 85 mg, about 86 mg,
about 87 mg, about 88 mg, about 89 mg, about 90 mg, about 91 mg,
about 92 mg, about 93 mg, about 94 mg, about 95 mg, about 96 mg,
about 97 mg, about 98 mg, about 99 mg, about 100 mg, about 101 mg,
about 102 mg, about 103 mg, about 104 mg, about 105 mg, about 106
mg, about 107 mg, about 108 mg, about 109 mg, about 110 mg, about
111 mg, about 112 mg, about 113 mg, about 114 mg, about 115 mg,
about 116 mg, about 117 mg, about 118 mg, about 119 mg, about 120
mg, about 121 mg, about 122 mg, about 123 mg, about 124 mg, about
125 mg, about 126 mg, about 127 mg, about 128 mg, about 129 mg,
about 130 mg, about 131 mg, about 132 mg, about 133 mg, about 134
mg, about 135 mg, about 136 mg, about 137 mg, about 138 mg, about
139 mg, about 140 mg; [0111] e) 75.0 mg, 75.1 mg, 75.2 mg, 75.3 mg,
75.4 mg, 75.5 mg, 75.6 mg, 75.7 mg, 75.8 mg, 75.9 mg, 76.0 mg, 76.1
mg, 76.2 mg, 76.3 mg. 76.4 mg, 76.5 mg, 76.6 mg, 76.7 mg, 76.8 mg,
76.9 mg, 77.0 mg, 77.1 mg, 77.2 mg, 77.3 mg, 77.4 mg, 77.5 mg, 77.6
mg, 77.7 mg, 77.8 mg, 77.9 mg, 78.0 mg, 78.1 mg, 78.2 mg, 78.3 mg.
78.4 mg, 78.5 mg, 78.6 mg, 78.7 mg, 78.8 mg, 78.9 mg, 79.0 mg, 79.1
mg, 79.2 mg, 79.3 mg, 79.4 mg, 79.5 mg, 79.6 mg, 79.7 mg, 79.8 mg,
79.9 mg, 80.0 mg, 80.1 mg, 80.2 mg, 80.3 mg. 80.4 mg, 80.5 mg, 80.6
mg, 80.7 mg, 80.8 mg, 80.9 mg, 81.0 mg, 81.1 mg, 81.2 mg, 81.3 mg,
81.4 mg, 81.5 mg, 81.6 mg, 81.7 mg, 81.8 mg, 81.9 mg, 82.0 mg, 82.1
mg, 82.2 mg, 82.3 mg. 82.4 mg, 82.5 mg, 82.6 mg, 82.7 mg, 82.8 mg,
82.9 mg, 83.0 mg, 83.1 mg, 83.2 mg, 83.3 mg, 83.4 mg, 83.5 mg, 83.6
mg, 83.7 mg, 83.8 mg, 83.9 mg, 84.0 mg, 84.1 mg, 84.2 mg, 84.3 mg.
84.4 mg, 84.5 mg, 84.6 mg, 84.7 mg, 84.8 mg, 84.9 mg, 85.0 mg; and
[0112] f) about 75.0 mg, about 75.1 mg, about 75.2 mg, about 75.3
mg, about 75.4 mg, about 75.5 mg, about 75.6 mg, about 75.7 mg,
about 75.8 mg, about 75.9 mg, about 76.0 mg, about 76.1 mg, about
76.2 mg, about 76.3 mg, about 76.4 mg, about 76.5 mg, about 76.6
mg, about 76.6 about 76.7 mg, about 76.8 mg, about 76.9 mg, about
77.0 mg, about 77.1 mg, about 77.2 mg, about 77.3 mg, about 77.4
mg, about 77.5 mg, about 77.6 mg, about 77.7 mg, about 77.8 mg,
about 77.9 mg, about 78.0 mg, about 78.1 mg, about 78.2 mg, about
78.3 mg. about 78.4 mg, about 78.5 mg, about 78.6 mg, about 78.7
mg, about 78.8 mg, about 78.9 mg, about 79.0 mg, about 79.1 mg,
about 79.2 mg, about 79.3 mg, about 79.4 mg, about 79.5 mg, about
79.6 mg, about 79.7 mg, about 79.8 mg, about 79.9 mg, about 80.0
mg, about 80.1 mg, about 80.2 mg, about 80.3 mg. about 80.4 mg,
about 80.5 mg, about 80.6 mg, about 80.7 mg, about 80.8 mg, about
80.9 mg, about 81.0 mg, about 81.1 mg, about 81.2 mg, about 81.3
mg, about 81.4 mg, about 81.5 mg, about 81.6 mg, about 81.7 mg,
about 81.8 mg, about 81.9 mg, about 82.0 mg, about 82.1 mg, about
82.2 mg, about 82.3 mg. about 82.4 mg, about 82.5 mg, about 82.6
mg, about 82.7 mg, about 82.8 mg, about 82.9 mg, about 83.0 mg,
about 83.1 mg, about 83.2 mg, about 83.3 mg, about 83.4 mg, about
83.5 mg, about 83.6 mg, about 83.7 mg, about 83.8 mg, about 83.9
mg, about 84.0 mg, about 84.1 mg, about 84.2 mg, about 84.3 mg.
about 84.4 mg, about 84.5 mg, about 84.6 mg, about 84.7 mg, about
84.8 mg, about 84.9 mg, and about 85.0 mg.
[0113] Embodiment 21. The pharmaceutical composition of embodiment
17, wherein the oligomeric compound is present in the dosage unit
at an amount selected from: less than about 300 mg, less than about
295 mg, less than about 290 mg, less than about 285 mg, less than
about 280 mg, less than about 275 mg, less than about 270 mg, less
than about 265 mg, less than about 260 mg, less than about 255 mg,
less than about 250 mg, less than about 245 mg, less than about 240
mg, less than about 235 mg, less than about 230 mg, less than about
225 mg, less than about 220 mg, less than about 215 mg, less than
about 210 mg, less than about 205 mg, less than about 200 mg, less
than about 195 mg, less than about 190 mg, less than about 185 mg,
less than about 180 mg, less than about 175 mg, less than about 170
mg, less than about 165 mg, less than about 160 mg, less than about
150 mg, less than about 145 mg, less than about 140 mg, less than
about 135 mg, less than about 130 mg, less than about 125 mg, less
than about 120 mg, less than about 115 mg, less than about 110 mg,
less than about 105 mg, less than about 100 mg, less than about 95
mg, less than about 90 mg, less than about 85 mg, less than about
80 mg, less than about 75 mg, less than about 70 mg, less than
about 65 mg, less than about 60 mg, less than about 55 mg, less
than about 50 mg, less than about 45 mg, less than about 40 mg,
less than about 35 mg, less than about 30 mg, less than about 25
mg, and less than about 20 mg.
[0114] Embodiment 22. The pharmaceutical composition of embodiment
19, wherein the oligomeric compound is present in the dosage unit
at an amount selected from: [0115] a) 10 mg to 140 mg, from 10 mg
to 130 mg, from 10 mg to 120 mg, from 10 mg to 110 mg, from 10 mg
to 100 mg, from 10 mg to 90 mg, from 10 mg to 80 mg, from 10 mg to
70 mg, from 10 mg to 60 mg, from 10 mg to 50 mg, from 10 mg to 40
mg, from 10 mg to 30 mg, from 10 mg to 20 mg, from 20 mg to 140 mg,
from 20 mg to 130 mg, from 20 mg to 120 mg, from 20 mg to 110 mg,
from 20 mg to 100 mg, from 20 mg to 90 mg, from 20 mg to 80 mg,
from 20 mg to 70 mg, from 20 mg to 60 mg, from 20 mg to 50 mg, from
20 mg to 40 mg, from 20 mg to 30 mg, from 30 mg to 140 mg, from 30
mg to 130 mg, from 30 mg to 120 mg, from 30 mg to 110 mg, from 30
mg to 100 mg, from 30 mg to 90 mg, from 30 mg to 80 mg, from 30 mg
to 70 mg, from 30 mg to 60 mg, from 30 mg to 50 mg, from 30 mg to
40 mg, from 40 mg to 140 mg, from 40 mg to 130 mg, from 40 mg to
120 mg, from 40 mg to 110 mg, from 40 mg to 100 mg, from 40 mg to
90 mg, from 40 mg to 80 mg, from 40 mg to 70 mg, from 40 mg to 60
mg, from 40 mg to 50 mg, from 50 mg to 140 mg, from 50 mg to 130
mg, from 50 mg to 120 mg, from 50 mg to 110 mg, from 50 mg to 100
mg, from 50 mg to 90 mg, from 50 mg to 80 mg, from 50 mg to 70 mg,
from 50 mg to 60 mg, from 60 mg to 140 mg, from 60 mg to 130 mg,
from 60 mg to 120 mg, from 60 mg to 110 mg, from 60 mg to 100 mg,
from 60 mg to 90 mg, from 60 mg to 80 mg, from 60 mg to 70 mg, from
70 mg to 140 mg, from 70 mg to 130 mg, from 70 mg to 120 mg, from
70 mg to 110 mg, from 70 mg to 100 mg, from 70 mg to 90 mg, from 70
mg to 80 mg, from 80 mg to 140 mg, from 80 mg to 130 mg, from 80 mg
to 120 mg, from 80 mg to 110 mg, from 80 mg to 100 mg, from 80 mg
to 90 mg, from 90 mg to 140 mg, from 90 mg to 130 mg, from 90 mg to
120 mg, from 90 mg to 110 mg, from 90 mg to 100 mg, from 100 mg to
140 mg, from 100 mg to 130 mg, from 100 mg to 120 mg, from 100 mg
to 110 mg, from 110 mg to 140 mg, from 110 mg to 130 mg, from 110
mg to 120 mg, from 120 mg to 140 mg, from 120 mg to 130 mg, from
130 mg to 140 mg, from 65 mg to 95 mg, from 65 mg to 90 mg, from 65
mg to 85 mg from 65 mg to 80 mg, from 65 mg to 75 mg, from 65 mg to
70 mg, from 70 mg to 95 mg, from 70 mg to 85 mg, from 70 mg to 75
mg, from 75 mg to 100 mg, from 75 mg to 95 mg, from 75 mg to 90 mg,
from 75 mg to 85 mg, from 75 mg to 80 mg, from 80 mg to 95 mg, from
80 mg to 85 mg, from 85 mg to 100 mg, from 85 mg to 90 mg, from 90
mg to 95 mg, from 95 mg to 100 mg, from 80 mg to 89 mg, from 80 mg
to 88 mg, from 80 mg to 87 mg, from 80 mg to 86 mg, from 80 mg to
84 mg, from 80 mg to 83 mg, from 80 mg to 82 mg, from 80 mg to 81
mg, from 81 mg to 90 mg, from 82 mg to 89 mg, from 82 mg to 88 mg,
from 82 mg to 87 mg, from 82 mg to 86 mg, from 82 mg to 85 mg, from
82 mg to 84 mg, from 82 mg to 83 mg, from 83 mg to 90 mg, from 83
mg to 89 mg, from 83 mg to 88 mg, from 83 mg to 87 mg, from 83 mg
to 86 mg, from 83 mg to 85 mg, from 83 mg to 84 mg, from 84 mg to
90 mg, from 84 mg to 89 mg, from 84 mg to 88 mg, from 84 mg to 87
mg, from 84 mg to 86 mg, from 84 mg to 85 mg, from 85 mg to 89 mg,
from 85 mg to 88 mg, from 85 mg to 87 mg, from 85 mg to 86 mg, from
86 mg to 90 mg, from 86 mg to 89 mg, from 86 mg to 88 mg, from 86
mg to 87 mg, from 87 mg to 90 mg, from 87 mg to 89 mg, from 87 mg
to 88 mg, from 88 mg to 90 mg, from 88 mg to 89 mg, from 89 mg to
90 mg; and [0116] b) less than about 300 mg, less than about 295
mg, less than about 290 mg, less than about 285 mg, less than about
280 mg, less than about 275 mg, less than 270 mg, less than 265 mg,
less than about 260 mg, less than 255 mg, less than about 250 mg,
less than about 245 mg, less than about 240 mg, less than about 235
mg, less than about 230 mg, less than about 225 mg, less than about
220 mg, less than about 215 mg, less than about 210 mg, less than
about 205 mg, less than about 200 mg, less than about 195 mg, less
than about 190 mg, less than about 185 mg, less than about 180 mg,
less than about 175 mg, less than about 170 mg, less than about 165
mg, less than about 160 mg, less than about 150 mg, less than about
145 mg, less than about 140 mg, less than about 135 mg, less than
about 130 mg, less than about 125 mg, less than about 120 mg, less
than about 115 mg, less than about 110 mg, less than about 105 mg,
less than about 100 mg, less than about 95 mg, less than about 90
mg, less than about 85 mg, less than about 80 mg, less than about
75 mg, less than about 70 mg, less than about 65 mg, less than
about 60 mg, less than about 55 mg, less than about 50 mg, less
than about 45 mg, less than about 40 mg, less than about 35 mg,
less than about 30 mg, less than about 25 mg, and less than about
20 mg.
[0117] Embodiment 23. The pharmaceutical composition of embodiment
19, wherein the oligomeric compound is present in the dosage unit
at an amount selected from: [0118] a) at least about 10 mg, at
least about 15 mg, at least about 20 mg, at least about 25 mg, at
least about 30 mg, at least about 35 mg, at least about 40 mg, at
least about 45 mg, at least about 50 mg, at least about 55 mg, at
least about 60 mg, at least about 65 mg, at least about 70 mg, at
least about 75 mg, at least about 80 mg, at least about 85 mg, at
least about 90 mg, at least about 95 mg, at least about 100 mg, at
least about 105 mg, at least about 115 mg, at least about 120 mg,
at least about 125 mg, at least about 130 mg, at least about 135
mg, at least about 140 mg, at least about 145 mg, at least about
150 mg; and [0119] b) at least 10 mg, at least 15 mg, at least 20
mg, at least 25 mg, at least 30 mg, at least 35 mg, at least 40 mg,
at least 45 mg, at least 50 mg, at least 55 mg, at least 60 mg, at
least 65 mg, at least 70 mg, at least 75 mg, at least 80 mg, at
least 85 mg, at least 90 mg, at least 95 mg, at least about 100 mg,
at least 105 mg, at least 115 mg, at least 120 mg, at least 125 mg,
at least 130 mg, at least 135 mg, at least 140 mg, at least 145 mg,
and at least 150 mg.
[0120] Embodiment 24. The pharmaceutical composition of any one of
embodiments 19-23, wherein the dosage unit has a volume selected
from: [0121] a) 0.1 ml to 1.5 ml, 0.1 ml to 1.4 ml, 0.1 ml to 1.3
ml, 0.1 ml to 1.2 ml, 0.1 ml to 1.1 ml, 0.1 ml to 1.0 ml, 0.1 ml to
0.9 ml, 0.1 ml to 0.8 ml, 0.1 ml to 0.7 ml, 0.1 ml to 0.6 ml, 0.1
ml to 0.5 ml, 0.1 ml to 0.4 ml, 0.1 ml to 0.3 ml, 0.1 ml to 0.2 ml,
0.2 ml to 1.5 ml, 0.2 ml to 1.4 ml, 0.2 ml to 1.3 ml, 0.2 ml to 1.2
ml, 0.2 ml to 1.1 ml, 0.2 ml to 1.0 ml, 0.2 ml to 0.9 ml, 0.2 ml to
0.8 ml, 0.2 ml to 0.7 ml, 0.2 ml to 0.6 ml, 0.2 ml to 0.5 ml, 0.2
ml to 0.4 ml, 0.2 ml to 0.3 ml, 0.3 ml to 1.5 ml, 0.3 ml to 1.4 ml,
0.3 ml to 1.3 ml, 0.3 ml to 1.2 ml, 0.3 ml to 1.1 ml, 0.3 ml to 1.0
ml, 0.3 ml to 0.9 ml, 0.3 ml to 0.8 ml, 0.3 ml to 0.7 ml, 0.3 ml to
0.6 ml, 0.3 ml to 0.5 ml, 0.3 ml to 0.4 ml, 0.4 ml to 1.5 ml, 0.4
ml to 1.4 ml, 0.4 ml to 1.3 ml, 0.4 ml to 1.2 ml, 0.4 ml to 1.1 ml,
0.4 ml to 1.0 ml, 0.4 ml to 0.9 ml, 0.4 ml to 0.8 ml, 0.4 ml to 0.7
ml, 0.4 ml to 0.6 ml, 0.4 ml to 0.5 ml, 0.5 ml to 1.5 ml, 0.5 ml to
1.4 ml, 0.5 ml to 1.3 ml, 0.5. ml to 1.2 ml, 0.5 ml to 1.1 ml, 0.5
ml to 1.0 ml, 0.5 ml to 0.9 ml, 0.5 ml to 0.8 ml, 0.5 ml to 0.7 ml,
0.5 ml to 0.6 ml, 0.6 ml to 1.5 ml, 0.6 ml to 1.4 ml, 0.6 ml to 1.3
ml, 0.6 ml to 1.2 ml, 0.6 ml to 1.1 ml, 0.6 ml to 1.0 ml, 0.6 ml to
0.9 ml, 0.6 ml to 0.8 ml, 0.6 ml to 0.7 ml, 0.7 ml, to 1.5 ml, 0.7
ml to 1.4 ml, 0.7 ml to 1.3 ml, 0.7 ml to 1.2 ml, 0.7 ml to 1.1 ml,
0.7 ml to 1.0 ml, 0.7 ml to 0.9 ml, 0.7 ml to 0.8 ml, 0.8 ml to 1.5
ml, 0.8 ml to 1.4 ml, 0.8 ml to 1.3 ml, 0.8 ml to 1.2 ml, 0.8 ml to
1.1 ml, 0.8 ml to 1.0 ml. 0.8 ml to 0.9 ml, 0.9 ml, to 1.5 ml, 0.9
ml to 1.4 ml, 0.9 ml to 1.3 ml, 0.9 ml to 1.2 ml, 0.9 ml, to 1.1
ml, 0.9 ml to 1.0 ml, 1.0 ml to 1.5 ml, 1.0 ml to 1.4 ml, 1.0 ml to
1.3 ml, 1.0 ml to 1.2 ml, 1.0 ml to 1.1 ml, 1.1 ml to 1.5 ml, 1.1
ml to 1.4 ml, 1.1 ml to 1.3 ml, 1.1 ml to 1.2 ml, 1.2 ml to 1.5 ml,
1.2 ml to 1.4 ml, 1.2 ml to 1.3 ml, 1.3 ml to 1.5 ml, 1.3 ml to 1.4
ml, 1.4 ml to, 1.5 ml; and [0122] b) about 0.1 ml to about 1.5 ml,
about 0.1 ml to about 1.4 ml, about 0.1 ml to about 1.3 ml, about
0.1 ml to about 1.2 ml, about 0.1 ml to about 1.1 ml, about 0.1 ml
to about 1.0 ml, about 0.1 ml to about 0.9 ml, about 0.1 ml to
about 0.8 ml, about 0.1 ml to about 0.7 ml, about 0.1 ml to about
0.6 ml, about 0.1 ml to about 0.5 ml, about 0.1 ml to about 0.4 ml,
about 0.1 ml to about 0.3 ml, about 0.1 ml to about 0.2 ml, about
0.2 ml to about 1.5 ml, about 0.2 ml to about 1.4 ml, about 0.2 ml
to about 1.3 ml, about 0.2 ml to about 1.2 ml, about 0.2 ml to
about 1.1 ml, about 0.2 ml to about 1.0 ml, about 0.2 ml to about
0.9 ml, about 0.2 ml to about 0.8 ml, about 0.2 ml to about 0.7 ml,
about 0.2 ml to about 0.6 ml, about 0.2 ml to about 0.5 ml, about
0.2 ml to about 0.4 ml, about 0.2 ml to about 0.3 ml, about 0.3 ml
to about 1.5 ml, about 0.3 ml to about 1.4 ml, about 0.3 ml to
about 1.3 ml, about 0.3 ml to about 1.2 ml, about 0.3 ml to about
1.1 ml, about 0.3 ml to about 1.0 ml, about 0.3 ml to about 0.9 ml,
about 0.3 ml to about 0.8 ml, about 0.3 ml to about 0.7 ml, about
0.3 ml to about 0.6 ml, about 0.3 ml to about 0.5 ml, about 0.3 ml
to about 0.4 ml, about 0.4 ml to about 1.5 ml, about 0.4 ml to
about 1.4 ml, about 0.4 ml to about 1.3 ml, about 0.4 ml to about
1.2 ml, about 0.4 ml to about 1.1 ml, about 0.4 ml to about 1.0 ml,
about 0.4 ml to about 0.9 ml, about 0.4 ml to about 0.8 ml, about
0.4 ml to about 0.7 ml, about 0.4 ml to about 0.6 ml, about 0.4 ml
to about 0.5 ml, about 0.5 ml to about 1.5 ml, about 0.5 ml to
about 1.4 ml, about 0.5 ml to about 1.3 ml, about 0.5. ml to about
1.2 ml, about 0.5 ml to about 1.1 ml, about 0.5 ml to about 1.0 ml,
about 0.5 ml to about 0.9 ml, about 0.5 ml to about 0.8 ml, about
0.5 ml to about 0.7 ml, about 0.5 ml to about 0.6 ml, about 0.6 ml
to about 1.5 ml, about 0.6 ml to about 1.4 ml, about 0.6 ml to
about 1.3 ml, about 0.6 ml to about 1.2 ml, about 0.6 ml to about
1.1 ml, about 0.6 ml to about 1.0 ml, about 0.6 ml to about 0.9 ml,
about 0.6 ml to about 0.8 ml, about 0.6 ml to about 0.7 ml, about
0.7 ml, about to about 1.5 ml, about 0.7 ml to about 1.4 ml, about
0.7 ml to about 1.3 ml, about 0.7 ml to about 1.2 ml, about 0.7 ml
to about 1.1 ml, about 0.7 ml to about 1.0 ml, about 0.7 ml to
about 0.9 ml, about 0.7 ml to about 0.8 ml, about 0.8 ml to about
1.5 ml, about 0.8 ml to about 1.4 ml, about 0.8 ml to about 1.3 ml,
about 0.8 ml to about 1.2 ml, about 0.8 ml to about 1.1 ml, about
0.8 ml to about 1.0 ml. 0.8 ml to about 0.9 ml, about 0.9 ml, about
to about 1.5 ml, about 0.9 ml to about 1.4 ml, about 0.9 ml to
about 1.3 ml, about 0.9 ml to about 1.2 ml, about 0.9 ml, about to
about 1.1 ml, about 0.9 ml to about 1.0 ml, about 1.0 ml to about
1.5 ml, about 1.0 ml to about 1.4 ml, about 1.0 ml to about 1.3 ml,
about 1.0 ml to about 1.2 ml, about 1.0 ml to about 1.1 ml, about
1.1 ml to about 1.5 ml, about 1.1 ml to about 1.4 ml, about 1.1 ml
to about 1.3 ml, about 1.1 ml to about 1.2 ml, about 1.2 ml to
about 1.5 ml, about 1.2 ml to about 1.4 ml, about 1.2 ml to about
1.3 ml, about 1.3 ml to about 1.5 ml, about 1.3 ml to about 1.4 ml,
and about 1.4 ml to about 1.5 ml.
[0123] Embodiment 25. The pharmaceutical composition of any one of
embodiments 14-24, wherein the pharmaceutical composition is
packaged in a pre-filled syringe.
[0124] Embodiment 26. A method comprising contacting a cell with
the oligomeric compound of any of embodiments 1-5.
[0125] Embodiment 27. A method comprising administering to an
animal a pharmaceutical composition comprising a therapeutically
effective amount of an oligomeric compound according to the
following formula:
##STR00005##
[0126] Embodiment 28. A method comprising administering to an
animal a therapeutically effective amount of the oligomeric
compound of any of embodiments 1-5 in the form of a pharmaceutical
composition.
[0127] Embodiment 29. The method of embodiment 27 or 28, wherein
the pharmaceutical composition consists or consists essentially of
the oligomeric compound and phosphate buffered saline.
[0128] Embodiment 30. A method comprising administering to an
animal the pharmaceutical compositions of any one of embodiments
14-25, wherein the pharmaceutical composition comprises a
therapeutically effective amount of the oligomeric compound.
[0129] Embodiment 31. The method of embodiment 30, wherein the
therapeutically effective amount does not significantly alter
platelet levels or platelet activity in the animal or does not
cause bleeding in the animal compared to an animal not administered
the pharmaceutical composition.
[0130] Embodiment 32. The method of any one of embodiments 27 to
31, comprising administering an amount of the oligomeric compound
selected from: [0131] a) 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg,
35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg, 65 mg, 70 mg, 75 mg, 80
mg, 85 mg, 90 mg, 95 mg, 100 mg, 105 mg, 110 mg, 115 mg, 120 mg,
125 mg, 130 mg, 135 mg, 140 mg, 145 mg, 150 mg, 155 mg, 160 mg, 165
mg, 170 mg, 175 mg, 180 mg, 185 mg, 190 mg, 195 mg, 200 mg, 205 mg,
210 mg, 215 mg, 220 mg, 225 mg, 230 mg, 235 mg, 240 mg, 245 mg, 250
mg, 255 mg, 260 mg, 265 mg, 270 mg, 275 mg, 280 mg, 285 mg, 290 mg,
295 mg, 300 mg; [0132] b) about 5 mg, about 10 mg, about 15 mg,
about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg,
about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg,
about 70 mg, about 75 mg, about 80 mg, about 85 mg, about 90 mg,
about 95 mg, about 100 mg, about 105 mg, about 110 mg, about 115
mg, about 120 mg, about 125 mg, about 130 mg, about 135 mg, about
140 mg, about 145 mg, about 150 mg, about 155 mg, about 160 mg,
about 165 mg, about 170 mg, about 175 mg, about 180 mg, about 185
mg, about 190 mg, about 195 mg, about 200 mg, about 205 mg, about
210 mg, about 215 mg, about 220 mg, about 225 mg, about 230 mg,
about 235 mg, about 240 mg, about 245 mg, about 250 mg, about 255
mg, about 260 mg, about 265 mg, about 270 mg, about 275 mg, about
280 mg, about 285 mg, about 290 mg, about 295 mg, about 300 mg;
[0133] c) 20 mg, 21 mg, 22 mg, 23 mg, 24 mg, 25 mg, 26 mg, 27 mg,
28 mg, 29 mg, 30 mg, 31 mg, 32 mg, 33 mg, 34 mg, 35 mg, 36 mg, 37
mg, 38 mg, 39 mg, 40 mg, 41 mg, 42 mg, 43 mg, 44 mg, 45 mg, 46 mg,
47 mg, 48 mg, 49 mg, 50 mg, 51 mg, 52 mg, 53 mg, 54 mg, 55 mg, 56
mg, 57 mg, 58 mg, 59 mg, 60 mg, 61 mg, 62 mg, 63 mg, 64 mg, 65 mg,
66 mg, 67 mg, 68 mg, 69 mg, 70 mg, 71 mg, 72 mg, 73 mg, 74 mg, 75
mg, 76 mg, 77 mg, 78 mg, 79 mg, 80 mg, 81 mg, 82 mg, 83 mg, 84 mg,
85 mg, 86 mg, 87 mg, 88 mg, 89 mg, 90 mg, 91 mg, 92 mg, 93 mg, 94
mg, 95 mg, 96 mg, 97 mg, 98 mg, 99 mg, 100 mg, 101 mg, 102 mg, 103
mg, 104 mg, 105 mg, 106 mg, 107 mg, 108 mg, 109 mg, 110 mg, 111 mg,
112 mg, 113 mg, 114 mg, 115 mg, 116 mg, 117 mg, 118 mg, 119 mg, 120
mg, 121 mg, 122 mg, 123 mg, 124 mg, 125 mg, 126 mg, 127 mg, 128 mg,
129 mg, 130 mg, 131 mg, 132 mg, 133 mg, 134 mg, 135 mg, 136 mg, 137
mg, 138 mg, 139 mg, 140 mg; [0134] d) about 20 mg, about 21 mg,
about 22 mg, about 23 mg, about 24 mg, about 25 mg, about 26 mg,
about 27 mg, about 28 mg, about 29 mg, about 30 mg, about 31 mg,
about 32 mg, about 33 mg, about 34 mg, about 35 mg, about 36 mg,
about 37 mg, about 38 mg, about 39 mg, about 40 mg, about 41 mg,
about 42 mg, about 43 mg, about 44 mg, about 45 mg, about 46 mg,
about 47 mg, about 48 mg, about 49 mg, about 50 mg, about 51 mg,
about 52 mg, about 53 mg, about 54 mg, about 55 mg, about 56 mg,
about 57 mg, about 58 mg, about 59 mg, about 60 mg, about 61 mg,
about 62 mg, about 63 mg, about 64 mg, about 65 mg, about 66 mg,
about 67 mg, about 68 mg, about 69 mg, about 70 mg, about 71 mg,
about 72 mg, about 73 mg, about 74 mg, about 75 mg, about 76 mg,
about 77 mg, about 78 mg, about 79 mg, about 80 mg, about 81 mg,
about 82 mg, about 83 mg, about 84 mg, about 85 mg, about 86 mg,
about 87 mg, about 88 mg, about 89 mg, about 90 mg, about 91 mg,
about 92 mg, about 93 mg, about 94 mg, about 95 mg, about 96 mg,
about 97 mg, about 98 mg, about 99 mg, about 100 mg, about 101 mg,
about 102 mg, about 103 mg, about 104 mg, about 105 mg, about 106
mg, about 107 mg, about 108 mg, about 109 mg, about 110 mg, about
111 mg, about 112 mg, about 113 mg, about 114 mg, about 115 mg,
about 116 mg, about 117 mg, about 118 mg, about 119 mg, about 120
mg, about 121 mg, about 122 mg, about 123 mg, about 124 mg, about
125 mg, about 126 mg, about 127 mg, about 128 mg, about 129 mg,
about 130 mg, about 131 mg, about 132 mg, about 133 mg, about 134
mg, about 135 mg, about 136 mg, about 137 mg, about 138 mg, about
139 mg, about 140 mg; [0135] e) 75.0 mg, 75.1 mg, 75.2 mg, 75.3 mg,
75.4 mg, 75.5 mg, 75.6 mg, 75.7 mg, 75.8 mg, 75.9 mg, 76.0 mg, 76.1
mg, 76.2 mg, 76.3 mg. 76.4 mg, 76.5 mg, 76.6 mg, 76.7 mg, 76.8 mg,
76.9 mg, 77.0 mg, 77.1 mg, 77.2 mg, 77.3 mg, 77.4 mg, 77.5 mg, 77.6
mg, 77.7 mg, 77.8 mg, 77.9 mg, 78.0 mg, 78.1 mg, 78.2 mg, 78.3 mg.
78.4 mg, 78.5 mg, 78.6 mg, 78.7 mg, 78.8 mg, 78.9 mg, 79.0 mg, 79.1
mg, 79.2 mg, 79.3 mg, 79.4 mg, 79.5 mg, 79.6 mg, 79.7 mg, 79.8 mg,
79.9 mg, 80.0 mg, 80.1 mg, 80.2 mg, 80.3 mg. 80.4 mg, 80.5 mg, 80.6
mg, 80.7 mg, 80.8 mg, 80.9 mg, 81.0 mg, 81.1 mg, 81.2 mg, 81.3 mg,
81.4 mg, 81.5 mg, 81.6 mg, 81.7 mg, 81.8 mg, 81.9 mg, 82.0 mg, 82.1
mg, 82.2 mg, 82.3 mg. 82.4 mg, 82.5 mg, 82.6 mg, 82.7 mg, 82.8 mg,
82.9 mg, 83.0 mg, 83.1 mg, 83.2 mg, 83.3 mg, 83.4 mg, 83.5 mg, 83.6
mg, 83.7 mg, 83.8 mg, 83.9 mg, 84.0 mg, 84.1 mg, 84.2 mg, 84.3 mg.
84.4 mg, 84.5 mg, 84.6 mg, 84.7 mg, 84.8 mg, 84.9 mg, 85.0 mg; and
[0136] f) about 75.0 mg, about 75.1 mg, about 75.2 mg, about 75.3
mg, about 75.4 mg, about 75.5 mg, about 75.6 mg, about 75.7 mg,
about 75.8 mg, about 75.9 mg, about 76.0 mg, about 76.1 mg, about
76.2 mg, about 76.3 mg, about 76.4 mg, about 76.5 mg, about 76.6
mg, about 76.6 about 76.7 mg, about 76.8 mg, about 76.9 mg, about
77.0 mg, about 77.1 mg, about 77.2 mg, about 77.3 mg, about 77.4
mg, about 77.5 mg, about 77.6 mg, about 77.7 mg, about 77.8 mg,
about 77.9 mg, about 78.0 mg, about 78.1 mg, about 78.2 mg, about
78.3 mg. about 78.4 mg, about 78.5 mg, about 78.6 mg, about 78.7
mg, about 78.8 mg, about 78.9 mg, about 79.0 mg, about 79.1 mg,
about 79.2 mg, about 79.3 mg, about 79.4 mg, about 79.5 mg, about
79.6 mg, about 79.7 mg, about 79.8 mg, about 79.9 mg, about 80.0
mg, about 80.1 mg, about 80.2 mg, about 80.3 mg. about 80.4 mg,
about 80.5 mg, about 80.6 mg, about 80.7 mg, about 80.8 mg, about
80.9 mg, about 81.0 mg, about 81.1 mg, about 81.2 mg, about 81.3
mg, about 81.4 mg, about 81.5 mg, about 81.6 mg, about 81.7 mg,
about 81.8 mg, about 81.9 mg, about 82.0 mg, about 82.1 mg, about
82.2 mg, about 82.3 mg. about 82.4 mg, about 82.5 mg, about 82.6
mg, about 82.7 mg, about 82.8 mg, about 82.9 mg, about 83.0 mg,
about 83.1 mg, about 83.2 mg, about 83.3 mg, about 83.4 mg, about
83.5 mg, about 83.6 mg, about 83.7 mg, about 83.8 mg, about 83.9
mg, about 84.0 mg, about 84.1 mg, about 84.2 mg, about 84.3 mg.
about 84.4 mg, about 84.5 mg, about 84.6 mg, about 84.7 mg, about
84.8 mg, about 84.9 mg, and about 85.0 mg.
[0137] Embodiment 33. The method of any one of embodiments 27 to
31, comprising administering an amount of the oligomeric compound
selected from: less than about 300 mg, less than about 295 mg, less
than about 290 mg, less than about 285 mg, less than about 280 mg,
less than about 275 mg, less than about 270 mg, less than about 265
mg, less than about 260 mg, less than about 255 mg, less than about
250 mg, less than about 245 mg, less than about 240 mg, less than
about 235 mg, less than about 230 mg, less than about 225 mg, less
than about 220 mg, less than about 215 mg, less than about 210 mg,
less than about 205 mg, less than about 200 mg, less than about 195
mg, less than about 190 mg, less than about 185 mg, less than about
180 mg, less than about 175 mg, less than about 170 mg, less than
about 165 mg, less than about 160 mg, less than about 150 mg, less
than about 145 mg, less than about 140 mg, less than about 135 mg,
less than about 130 mg, less than about 125 mg, less than about 120
mg, less than about 115 mg, less than about 110 mg, less than about
105 mg, less than about 100 mg, less than about 95 mg, less than
about 90 mg, less than about 85 mg, less than about 80 mg, less
than about 75 mg, less than about 70 mg, less than about 65 mg,
less than about 60 mg, less than about 55 mg, less than about 50
mg, less than about 45 mg, less than about 40 mg, less than about
35 mg, less than about 30 mg, less than about 25 mg, and less than
about 20 mg.
[0138] Embodiment 34. The method of any one of embodiments 27 to
31, comprising administering an amount of the oligomeric compound
selected from: [0139] a) 10 mg to 140 mg, from 10 mg to 130 mg,
from 10 mg to 120 mg, from 10 mg to 110 mg, from 10 mg to 100 mg,
from 10 mg to 90 mg, from 10 mg to 80 mg, from 10 mg to 70 mg, from
10 mg to 60 mg, from 10 mg to 50 mg, from 10 mg to 40 mg, from 10
mg to 30 mg, from 10 mg to 20 mg, from 20 mg to 140 mg, from 20 mg
to 130 mg, from 20 mg to 120 mg, from 20 mg to 110 mg, from 20 mg
to 100 mg, from 20 mg to 90 mg, from 20 mg to 80 mg, from 20 mg to
70 mg, from 20 mg to 60 mg, from 20 mg to 50 mg, from 20 mg to 40
mg, from 20 mg to 30 mg, from 30 mg to 140 mg, from 30 mg to 130
mg, from 30 mg to 120 mg, from 30 mg to 110 mg, from 30 mg to 100
mg, from 30 mg to 90 mg, from 30 mg to 80 mg, from 30 mg to 70 mg,
from 30 mg to 60 mg, from 30 mg to 50 mg, from 30 mg to 40 mg, from
40 mg to 140 mg, from 40 mg to 130 mg, from 40 mg to 120 mg, from
40 mg to 110 mg, from 40 mg to 100 mg, from 40 mg to 90 mg, from 40
mg to 80 mg, from 40 mg to 70 mg, from 40 mg to 60 mg, from 40 mg
to 50 mg, from 50 mg to 140 mg, from 50 mg to 130 mg, from 50 mg to
120 mg, from 50 mg to 110 mg, from 50 mg to 100 mg, from 50 mg to
90 mg, from 50 mg to 80 mg, from 50 mg to 70 mg, from 50 mg to 60
mg, from 60 mg to 140 mg, from 60 mg to 130 mg, from 60 mg to 120
mg, from 60 mg to 110 mg, from 60 mg to 100 mg, from 60 mg to 90
mg, from 60 mg to 80 mg, from 60 mg to 70 mg, from 70 mg to 140 mg,
from 70 mg to 130 mg, from 70 mg to 120 mg, from 70 mg to 110 mg,
from 70 mg to 100 mg, from 70 mg to 90 mg, from 70 mg to 80 mg,
from 80 mg to 140 mg, from 80 mg to 130 mg, from 80 mg to 120 mg,
from 80 mg to 110 mg, from 80 mg to 100 mg, from 80 mg to 90 mg,
from 90 mg to 140 mg, from 90 mg to 130 mg, from 90 mg to 120 mg,
from 90 mg to 110 mg, from 90 mg to 100 mg, from 100 mg to 140 mg,
from 100 mg to 130 mg, from 100 mg to 120 mg, from 100 mg to 110
mg, from 110 mg to 140 mg, from 110 mg to 130 mg, from 110 mg to
120 mg, from 120 mg to 140 mg, from 120 mg to 130 mg, from 130 mg
to 140 mg, from 65 mg to 95 mg, from 65 mg to 90 mg, from 65 mg to
85 mg from 65 mg to 80 mg, from 65 mg to 75 mg, from 65 mg to 70
mg, from 70 mg to 95 mg, from 70 mg to 85 mg, from 70 mg to 75 mg,
from 75 mg to 100 mg, from 75 mg to 95 mg, from 75 mg to 90 mg,
from 75 mg to 85 mg, from 75 mg to 80 mg, from 80 mg to 95 mg, from
80 mg to 85 mg, from 85 mg to 100 mg, from 85 mg to 90 mg, from 90
mg to 95 mg, from 95 mg to 100 mg, from 80 mg to 89 mg, from 80 mg
to 88 mg, from 80 mg to 87 mg, from 80 mg to 86 mg, from 80 mg to
84 mg, from 80 mg to 83 mg, from 80 mg to 82 mg, from 80 mg to 81
mg, from 81 mg to 90 mg, from 82 mg to 89 mg, from 82 mg to 88 mg,
from 82 mg to 87 mg, from 82 mg to 86 mg, from 82 mg to 85 mg, from
82 mg to 84 mg, from 82 mg to 83 mg, from 83 mg to 90 mg, from 83
mg to 89 mg, from 83 mg to 88 mg, from 83 mg to 87 mg, from 83 mg
to 86 mg, from 83 mg to 85 mg, from 83 mg to 84 mg, from 84 mg to
90 mg, from 84 mg to 89 mg, from 84 mg to 88 mg, from 84 mg to 87
mg, from 84 mg to 86 mg, from 84 mg to 85 mg, from 85 mg to 89 mg,
from 85 mg to 88 mg, from 85 mg to 87 mg, from 85 mg to 86 mg, from
86 mg to 90 mg, from 86 mg to 89 mg, from 86 mg to 88 mg, from 86
mg to 87 mg, from 87 mg to 90 mg, from 87 mg to 89 mg, from 87 mg
to 88 mg, from 88 mg to 90 mg, from 88 mg to 89 mg, from 89 mg to
90 mg; and [0140] b) less than about 300 mg, less than about 295
mg, less than about 290 mg, less than about 285 mg, less than about
280 mg, less than about 275 mg, less than 270 mg, less than 265 mg,
less than about 260 mg, less than 255 mg, less than about 250 mg,
less than about 245 mg, less than about 240 mg, less than about 235
mg, less than about 230 mg, less than about 225 mg, less than about
220 mg, less than about 215 mg, less than about 210 mg, less than
about 205 mg, less than about 200 mg, less than about 195 mg, less
than about 190 mg, less than about 185 mg, less than about 180 mg,
less than about 175 mg, less than about 170 mg, less than about 165
mg, less than about 160 mg, less than about 150 mg, less than about
145 mg, less than about 140 mg, less than about 135 mg, less than
about 130 mg, less than about 125 mg, less than about 120 mg, less
than about 115 mg, less than about 110 mg, less than about 105 mg,
less than about 100 mg, less than about 95 mg, less than about 90
mg, less than about 85 mg, less than about 80 mg, less than about
75 mg, less than about 70 mg, less than about 65 mg, less than
about 60 mg, less than about 55 mg, less than about 50 mg, less
than about 45 mg, less than about 40 mg, less than about 35 mg,
less than about 30 mg, less than about 25 mg, and less than about
20 mg.
[0141] Embodiment 35. The method of any one of embodiments 27 to
31, comprising administering an amount of the oligomeric compound
selected from: [0142] a) at least about 10 mg, at least about 15
mg, at least about 20 mg, at least about 25 mg, at least about 30
mg, at least about 35 mg, at least about 40 mg, at least about 45
mg, at least about 50 mg, at least about 55 mg, at least about 60
mg, at least about 65 mg, at least about 70 mg, at least about 75
mg, at least about 80 mg, at least about 85 mg, at least about 90
mg, at least about 95 mg, at least about 100 mg, at least about 105
mg, at least about 115 mg, at least about 120 mg, at least about
125 mg, at least about 130 mg, at least about 135 mg, at least
about 140 mg, at least about 145 mg, at least about 150 mg; and
[0143] b) at least 10 mg, at least 15 mg, at least 20 mg, at least
25 mg, at least 30 mg, at least 35 mg, at least 40 mg, at least 45
mg, at least 50 mg, at least 55 mg, at least 60 mg, at least 65 mg,
at least 70 mg, at least 75 mg, at least 80 mg, at least 85 mg, at
least 90 mg, at least 95 mg, at least about 100 mg, at least 105
mg, at least 115 mg, at least 120 mg, at least 125 mg, at least 130
mg, at least 135 mg, at least 140 mg, at least 145 mg, and at least
150 mg.
[0144] Embodiment 36. The method of any one of embodiments 27 to
35, wherein the concentration of the oligomeric compound in the
pharmaceutically acceptable carrier or diluent is selected from:
[0145] a) 5 mg/ml, 10 mg/ml, 15 mg/ml, 20 mg/ml, 25 mg/ml, 30
mg/ml, 35 mg/ml, 40 mg/ml, 45 mg/ml, 50 mg/ml, 55 mg/ml, 60 mg/ml,
65 mg/ml, 70 mg/ml, 75 mg/ml, 80 mg/ml, 85 mg/ml, 90 mg/ml, 95
mg/ml, 100 mg/ml, 105 mg/ml, 110 mg/ml, 115 mg/ml, 120 mg/ml, 125
mg/ml, 130 mg/ml, 135 mg/ml, 140 mg/ml, 145 mg/ml, 150 mg/ml, 155
mg/ml, 160 mg/ml; [0146] b) about 5 mg/ml, about 10 mg/ml, about 15
mg/ml, about 20 mg/ml, about 25 mg/ml, about 30 mg/ml, about 35
mg/ml, about 40 mg/ml, about 45 mg/ml, about 50 mg/ml, about 55
mg/ml, about 60 mg/ml, about 65 mg/ml, about 70 mg/ml, about 75
mg/ml, about 80 mg/ml, about 85 mg/ml, about 90 mg/ml, about 95
mg/ml, about 100 mg/ml, about 105 mg/ml, about 110 mg/ml, about 115
mg/ml, about 120 mg/ml, about 125 mg/ml, about 130 mg/ml, about 135
mg/ml, about 140 mg/ml, about 145 mg/ml, about 150 mg/ml, about 155
mg/ml, about 160 mg/ml; and [0147] c) 20 mg/ml, 21 mg/ml, 22 mg/ml,
23 mg/ml, 24 mg/ml, 25 mg/ml, 26 mg/ml, 27 mg/ml, 28 mg/ml, 29
mg/ml, 30 mg/ml, 31 mg/ml, 32 mg/ml, 33 mg/ml, 34 mg/ml, 35 mg/ml,
36 mg/ml, 37 mg/ml, 38 mg/ml, 39 mg/ml, 40 mg/ml, 41 mg/ml, 42
mg/ml, 43 mg/ml, 44 mg/ml, 45 mg/ml, 46 mg/ml, 47 mg/ml, 48 mg/ml,
49 mg/ml, 50 mg/ml, 51 mg/ml, 52 mg/ml, 53 mg/ml, 54 mg/ml, 55
mg/ml, 56 mg/ml, 57 mg/ml, 58 mg/ml, 59 mg/ml, 60 mg/ml, 61 mg/ml,
62 mg/ml, 63 mg/ml, 64 mg/ml, 65 mg/ml, 66 mg/ml, 67 mg/ml, 68
mg/ml, 69 mg/ml, 70 mg/ml, 71 mg/ml, 72 mg/ml, 73 mg/ml, 74 mg/ml,
75 mg/ml, 76 mg/ml, 77 mg/ml, 78 mg/ml, 79 mg/ml, 80 mg/ml, 81
mg/ml, 82 mg/ml, 83 mg/ml, 84 mg/ml, 85 mg/ml, 86 mg/ml, 87 mg/ml,
88 mg/ml, 89 mg/ml, 90 mg/ml, 91 mg/ml, 92 mg/ml, 93 mg/ml, 94
mg/ml, 95 mg/ml, 96 mg/ml, 97 mg/ml, 98 mg/ml, 99 mg/ml, 100 mg/ml,
101 mg/ml, 102 mg/ml, 103 mg/ml, 104 mg/ml, 105 mg/ml, 106 mg/ml,
107 mg/ml, 108 mg/ml, 109 mg/ml, 110 mg/ml, 111 mg/ml, 112 mg/ml,
113 mg/ml, 114 mg/ml, 115 mg/ml, 116 mg/ml, 117 mg/ml, 118 mg/ml,
119 mg/ml, 120 mg/ml, 121 mg/ml, 122 mg/ml, 123 mg/ml, 124 mg/ml,
125 mg/ml, 126 mg/ml, 127 mg/ml, 128 mg/ml, 129 mg/ml, 130 mg/ml,
131 mg/ml, 132 mg/ml, 133 mg/ml, 134 mg/ml, 135 mg/ml, 136 mg/ml,
137 mg/ml, 138 mg/ml, 139 mg/ml, 140 mg/ml; [0148] d) about 20
mg/ml, about 21 mg/ml, about 22 mg/ml, about 23 mg/ml, about 24
mg/ml, about 25 mg/ml, about 26 mg/ml, about 27 mg/ml, about 28
mg/ml, about 29 mg/ml, about 30 mg/ml, about 31 mg/ml, about 32
mg/ml, about 33 mg/ml, about 34 mg/ml, about 35 mg/ml, about 36
mg/ml, about 37 mg/ml, about 38 mg/ml, about 39 mg/ml, about 40
mg/ml, about 41 mg/ml, about 42 mg/ml, about 43 mg/ml, about 44
mg/ml, about 45 mg/ml, about 46 mg/ml, about 47 mg/ml, about 48
mg/ml, about 49 mg/ml, about 50 mg/ml, about 51 mg/ml, about 52
mg/ml, about 53 mg/ml, about 54 mg/ml, about 55 mg/ml, about 56
mg/ml, about 57 mg/ml, about 58 mg/ml, about 59 mg/ml, about 60
mg/ml, about 61 mg/ml, about 62 mg/ml, about 63 mg/ml, about 64
mg/ml, about 65 mg/ml, about 66 mg/ml, about 67 mg/ml, about 68
mg/ml, about 69 mg/ml, about 70 mg/ml, about 71 mg/ml, about 72
mg/ml, about 73 mg/ml, about 74 mg/ml, about 75 mg/ml, about 76
mg/ml, about 77 mg/ml, about 78 mg/ml, about 79 mg/ml, about 80
mg/ml, about 81 mg/ml, about 82 mg/ml, about 83 mg/ml, about 84
mg/ml, about 85 mg/ml, about 86 mg/ml, about 87 mg/ml, about 88
mg/ml, about 89 mg/ml, about 90 mg/ml, about 91 mg/ml, about 92
mg/ml, about 93 mg/ml, about 94 mg/ml, about 95 mg/ml, about 96
mg/ml, about 97 mg/ml, about 98 mg/ml, about 99 mg/ml, about 100
mg/ml, about 101 mg/ml, about 102 mg/ml, about 103 mg/ml, about 104
mg/ml, about 105 mg/ml, about 106 mg/ml, about 107 mg/ml, about 108
mg/ml, about 109 mg/ml, about 110 mg/ml, about 111 mg/ml, about 112
mg/ml, about 113 mg/ml, about 114 mg/ml, about 115 mg/ml, about 116
mg/ml, about 117 mg/ml, about 118 mg/ml, about 119 mg/ml, about 120
mg/ml, about 121 mg/ml, about 122 mg/ml, about 123 mg/ml, about 124
mg/ml, about 125 mg/ml, about 126 mg/ml, about 127 mg/ml, about 128
mg/ml, about 129 mg/ml, about 130 mg/ml, about 131 mg/ml, about 132
mg/ml, about 133 mg/ml, about 134 mg/ml, about 135 mg/ml, about 136
mg/ml, about 137 mg/ml, about 138 mg/ml, about 139 mg/ml, and about
140 mg/ml.
[0149] Embodiment 37. The method of any one of embodiments 27 to
35, wherein the concentration of the oligomeric compound in the
pharmaceutically acceptable carrier or diluent is selected from: 20
mg/ml to 180 mg/ml, 20 mg/ml to 170 mg, 20 mg/ml to 160 mg/ml, 20
mg/ml to 150 mg/ml, 20 mg/ml to 140 mg/ml, 20 mg/ml to 130 mg/ml,
20 mg/ml to 120 mg/ml, 20 mg/ml to 110 mg/ml, 20 mg/ml to 100
mg/ml, 20 mg/ml to 90 mg/ml, 20 mg/ml to 80 mg/ml, 20 mg/ml to 70
mg/ml, 20 mg/ml to 60 mg/ml, 20 mg/ml to 50 mg/ml, 20 mg/ml to 40
mg/ml, 20 mg/ml to 30 mg/ml, 30 mg/ml to 180 mg/ml, 30 mg/ml to 170
mg, 30 mg/ml to 160 mg/ml, 30 mg/ml to 150 mg/ml, 30 mg/ml to 140
mg/ml, 30 mg/ml to 130 mg/ml, 30 mg/ml to 120 mg/ml, 30 mg/ml to
110 mg/ml, 30 mg/ml to 100 mg/ml, 30 mg/ml to 90 mg/ml, 30 mg/ml to
80 mg/ml, 30 mg/ml to 70 mg/ml, 30 mg/ml to 60 mg/ml, 30 mg/ml to
50 mg/ml, 30 mg/ml to 40 mg/ml, 40 mg/ml to 180 mg/ml, 40 mg/ml to
170 mg, 40 mg/ml to 160 mg/ml, 40 mg/ml to 150 mg/ml, 40 mg/ml to
140 mg/ml, 40 mg/ml to 130 mg/ml, 40 mg/ml to 120 mg/ml, 40 mg/ml
to 110 mg/ml, 40 mg/ml to 100 mg/ml, 40 mg/ml to 90 mg/ml, 40 mg/ml
to 80 mg/ml, 40 mg/ml to 70 mg/ml, 40 mg/ml to 60 mg/ml, 40 mg/ml
to 50 mg/ml, 50 mg/ml to 180 mg/ml, 50 mg/ml to 170 mg, 50 mg/ml to
160 mg/ml, 50 mg/ml to 150 mg/ml, 50 mg/ml to 140 mg/ml, 50 mg/ml
to 130 mg/ml, 50 mg/ml to 120 mg/ml, 50 mg/ml to 110 mg/ml, 50
mg/ml to 100 mg/ml, 50 mg/ml to 90 mg/ml, 50 mg/ml to 80 mg/ml, 50
mg/ml to 70 mg/ml, 50 mg/ml to 60 mg/ml, 60 mg/ml to 180 mg/ml, 60
mg/ml to 170 mg, 60 mg/ml to 160 mg/ml, 60 mg/ml to 150 mg/ml, 60
mg/ml to 140 mg/ml, 60 mg/ml to 130 mg/ml, 60 mg/ml to 120 mg/ml,
60 mg/ml to 110 mg/ml, 60 mg/ml to 100 mg/ml, 60 mg/ml to 90 mg/ml,
60 mg/ml to 80 mg/ml, 60 mg/ml to 70 mg/ml, 70 mg/ml to 180 mg/ml,
70 mg/ml to 170 mg, 70 mg/ml to 160 mg/ml, 70 mg/ml to 150 mg/ml,
70 mg/ml to 140 mg/ml, 70 mg/ml to 130 mg/ml, 70 mg/ml to 120
mg/ml, 70 mg/ml to 110 mg/ml, 70 mg/ml to 100 mg/ml, 70 mg/ml to 90
mg/ml, 70 mg/ml to 80 mg/ml, 80 mg/ml to 180 mg/ml, 80 mg/ml to 170
mg, 80 mg/ml to 160 mg/ml, 80 mg/ml to 150 mg/ml, 80 mg/ml to 140
mg/ml, 80 mg/ml to 130 mg/ml, 80 mg/ml to 120 mg/ml, 80 mg/ml to
110 mg/ml, 80 mg/ml to 100 mg/ml, 80 mg/ml to 90 mg/ml, 90 mg/ml to
180 mg/ml, 90 mg/ml to 170 mg, 90 mg/ml to 160 mg/ml, 90 mg/ml to
150 mg/ml, 90 mg/ml to 140 mg/ml, 90 mg/ml to 130 mg/ml, 90 mg/ml
to 120 mg/ml, 90 mg/ml to 110 mg/ml, 90 mg/ml to 100 mg/ml, 100
mg/ml to 180 mg/ml, 100 mg/ml to 170 mg, 100 mg/ml to 160 mg/ml,
100 mg/ml to 150 mg/ml, 100 mg/ml to 140 mg/ml, 100 mg/ml to 130
mg/ml, 100 mg/ml to 120 mg/ml, 100 mg/ml to 110 mg/ml, 110 mg/ml to
180 mg/ml, 110 mg/ml to 170 mg, 110 mg/ml to 160 mg/ml, 110 mg/ml
to 150 mg/ml, 110 mg/ml to 140 mg/ml, 110 mg/ml to 130 mg/ml, 110
mg/ml to 120 mg/ml, 120 mg/ml to 180 mg/ml, 120 mg/ml to 170 mg,
120 mg/ml to 160 mg/ml, 120 mg/ml to 150 mg/ml, 120 mg/ml to 140
mg/ml, 120 mg/ml to 130 mg/ml, 130 mg/ml to 180 mg/ml, 130 mg/ml to
170 mg, 130 mg/ml to 160 mg/ml, 130 mg/ml to 150 mg/ml, 130 mg/ml
to 140 mg/ml, 140 mg/ml to 180 mg/ml, 140 mg/ml to 170 mg/ml, 140
mg/ml to 160 mg/ml, 140 mg/ml to 150 mg/ml, 150 mg/ml to 180 mg/ml,
150 mg/ml to 170 mg/ml, 150 mg/ml to 160 mg/ml, 160 mg/ml to 180
mg/ml, 160 mg/ml to 170 mg/ml, and 170 mg/ml to 180 mg/ml.
[0150] Embodiment 38. The method of any one of embodiments 27 to
37, wherein the pharmaceutical composition is a form of a dosage
unit, wherein the dosage unit is characterized by a volume selected
from: [0151] a) 0.1 ml to 1.5 ml, 0.1 ml to 1.4 ml, 0.1 ml to 1.3
ml, 0.1 ml to 1.2 ml, 0.1 ml to 1.1 ml, 0.1 ml to 1.0 ml, 0.1 ml to
0.9 ml, 0.1 ml to 0.8 ml, 0.1 ml to 0.7 ml, 0.1 ml to 0.6 ml, 0.1
ml to 0.5 ml, 0.1 ml to 0.4 ml, 0.1 ml to 0.3 ml, 0.1 ml to 0.2 ml,
0.2 ml to 1.5 ml, 0.2 ml to 1.4 ml, 0.2 ml to 1.3 ml, 0.2 ml to 1.2
ml, 0.2 ml to 1.1 ml, 0.2 ml to 1.0 ml, 0.2 ml to 0.9 ml, 0.2 ml to
0.8 ml, 0.2 ml to 0.7 ml, 0.2 ml to 0.6 ml, 0.2 ml to 0.5 ml, 0.2
ml to 0.4 ml, 0.2 ml to 0.3 ml, 0.3 ml to 1.5 ml, 0.3 ml to 1.4 ml,
0.3 ml to 1.3 ml, 0.3 ml to 1.2 ml, 0.3 ml to 1.1 ml, 0.3 ml to 1.0
ml, 0.3 ml to 0.9 ml, 0.3 ml to 0.8 ml, 0.3 ml to 0.7 ml, 0.3 ml to
0.6 ml, 0.3 ml to 0.5 ml, 0.3 ml to 0.4 ml, 0.4 ml to 1.5 ml, 0.4
ml to 1.4 ml, 0.4 ml to 1.3 ml, 0.4 ml to 1.2 ml, 0.4 ml to 1.1 ml,
0.4 ml to 1.0 ml, 0.4 ml to 0.9 ml, 0.4 ml to 0.8 ml, 0.4 ml to 0.7
ml, 0.4 ml to 0.6 ml, 0.4 ml to 0.5 ml, 0.5 ml to 1.5 ml, 0.5 ml to
1.4 ml, 0.5 ml to 1.3 ml, 0.5, ml to 1.2 ml, 0.5 ml to 1.1 ml, 0.5
ml to 1.0 ml, 0.5 ml to 0.9 ml, 0.5 ml to 0.8 ml, 0.5 ml to 0.7 ml,
0.5 ml to 0.6 ml, 0.6 ml to 1.5 ml, 0.6 ml to 1.4 ml, 0.6 ml to 1.3
ml, 0.6 ml to 1.2 ml, 0.6 ml to 1.1 ml, 0.6 ml to 1.0 ml, 0.6 ml to
0.9 ml, 0.6 ml to 0.8 ml, 0.6 ml to 0.7 ml, 0.7 ml, to 1.5 ml, 0.7
ml to 1.4 ml, 0.7 ml to 1.3 ml, 0.7 ml to 1.2 ml, 0.7 ml to 1.1 ml,
0.7 ml to 1.0 ml, 0.7 ml to 0.9 ml, 0.7 ml to 0.8 ml, 0.8 ml to 1.5
ml, 0.8 ml to 1.4 ml, 0.8 ml to 1.3 ml, 0.8 ml to 1.2 ml, 0.8 ml to
1.1 ml, 0.8 ml to 1.0 ml. 0.8 ml to 0.9 ml, 0.9 ml, to 1.5 ml, 0.9
ml to 1.4 ml, 0.9 ml to 1.3 ml, 0.9 ml to 1.2 ml, 0.9 ml, to 1.1
ml, 0.9 ml to 1.0 ml, 1.0 ml to 1.5 ml, 1.0 ml to 1.4 ml, 1.0 ml to
1.3 ml, 1.0 ml to 1.2 ml, 1.0 ml to 1.1 ml, 1.1 ml to 1.5 ml, 1.1
ml to 1.4 ml, 1.1 ml to 1.3 ml, 1.1 ml to 1.2 ml, 1.2 ml to 1.5 ml,
1.2 ml to 1.4 ml, 1.2 ml to 1.3 ml, 1.3 ml to 1.5 ml, 1.3 ml to 1.4
ml, 1.4 ml to, 1.5 ml; and [0152] b) about 0.1 ml to about 1.5 ml,
about 0.1 ml to about 1.4 ml, about 0.1 ml to about 1.3 ml, about
0.1 ml to about 1.2 ml, about 0.1 ml to about 1.1 ml, about 0.1 ml
to about 1.0 ml, about 0.1 ml to about 0.9 ml, about 0.1 ml to
about 0.8 ml, about 0.1 ml to about 0.7 ml, about 0.1 ml to about
0.6 ml, about 0.1 ml to about 0.5 ml, about 0.1 ml to about 0.4 ml,
about 0.1 ml to about 0.3 ml, about 0.1 ml to about 0.2 ml, about
0.2 ml to about 1.5 ml, about 0.2 ml to about 1.4 ml, about 0.2 ml
to about 1.3 ml, about 0.2 ml to about 1.2 ml, about 0.2 ml to
about 1.1 ml, about 0.2 ml to about 1.0 ml, about 0.2 ml to about
0.9 ml, about 0.2 ml to about 0.8 ml, about 0.2 ml to about 0.7 ml,
about 0.2 ml to about 0.6 ml, about 0.2 ml to about 0.5 ml, about
0.2 ml to about 0.4 ml, about 0.2 ml to about 0.3 ml, about 0.3 ml
to about 1.5 ml, about 0.3 ml to about 1.4 ml, about 0.3 ml to
about 1.3 ml, about 0.3 ml to about 1.2 ml, about 0.3 ml to about
1.1 ml, about 0.3 ml to about 1.0 ml, about 0.3 ml to about 0.9 ml,
about 0.3 ml to about 0.8 ml, about 0.3 ml to about 0.7 ml, about
0.3 ml to about 0.6 ml, about 0.3 ml to about 0.5 ml, about 0.3 ml
to about 0.4 ml, about 0.4 ml to about 1.5 ml, about 0.4 ml to
about 1.4 ml, about 0.4 ml to about 1.3 ml, about 0.4 ml to about
1.2 ml, about 0.4 ml to about 1.1 ml, about 0.4 ml to about 1.0 ml,
about 0.4 ml to about 0.9 ml, about 0.4 ml to about 0.8 ml, about
0.4 ml to about 0.7 ml, about 0.4 ml to about 0.6 ml, about 0.4 ml
to about 0.5 ml, about 0.5 ml to about 1.5 ml, about 0.5 ml to
about 1.4 ml, about 0.5 ml to about 1.3 ml, about 0.5. ml to about
1.2 ml, about 0.5 ml to about 1.1 ml, about 0.5 ml to about 1.0 ml,
about 0.5 ml to about 0.9 ml, about 0.5 ml to about 0.8 ml, about
0.5 ml to about 0.7 ml, about 0.5 ml to about 0.6 ml, about 0.6 ml
to about 1.5 ml, about 0.6 ml to about 1.4 ml, about 0.6 ml to
about 1.3 ml, about 0.6 ml to about 1.2 ml, about 0.6 ml to about
1.1 ml, about 0.6 ml to about 1.0 ml, about 0.6 ml to about 0.9 ml,
about 0.6 ml to about 0.8 ml, about 0.6 ml to about 0.7 ml, about
0.7 ml, about to about 1.5 ml, about 0.7 ml to about 1.4 ml, about
0.7 ml to about 1.3 ml, about 0.7 ml to about 1.2 ml, about 0.7 ml
to about 1.1 ml, about 0.7 ml to about 1.0 ml, about 0.7 ml to
about 0.9 ml, about 0.7 ml to about 0.8 ml, about 0.8 ml to about
1.5 ml, about 0.8 ml to about 1.4 ml, about 0.8 ml to about 1.3 ml,
about 0.8 ml to about 1.2 ml, about 0.8 ml to about 1.1 ml, about
0.8 ml to about 1.0 ml. 0.8 ml to about 0.9 ml, about 0.9 ml, about
to about 1.5 ml, about 0.9 ml to about 1.4 ml, about 0.9 ml to
about 1.3 ml, about 0.9 ml to about 1.2 ml, about 0.9 ml, about to
about 1.1 ml, about 0.9 ml to about 1.0 ml, about 1.0 ml to about
1.5 ml, about 1.0 ml to about 1.4 ml, about 1.0 ml to about 1.3 ml,
about 1.0 ml to about 1.2 ml, about 1.0 ml to about 1.1 ml, about
1.1 ml to about 1.5 ml, about 1.1 ml to about 1.4 ml, about 1.1 ml
to about 1.3 ml, about 1.1 ml to about 1.2 ml, about 1.2 ml to
about 1.5 ml, about 1.2 ml to about 1.4 ml, about 1.2 ml to about
1.3 ml, about 1.3 ml to about 1.5 ml, about 1.3 ml to about 1.4 ml,
and about 1.4 ml to about 1.5 ml.
[0153] Embodiment 39. The method of any one of embodiments 27-38,
comprising administering a first dose and a second dose of the
pharmaceutical composition.
[0154] Embodiment 40. The method of embodiment 39, wherein the
first dose and the second dose are separated by 5, 10, 15, 20, 25,
30, 35, or 40 days.
[0155] Embodiment 41. The method of embodiment 39, wherein the
first dose and the second dose are separated by 20, 21, 22, 23, 24,
25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40
days.
[0156] Embodiment 42. The method of embodiment 39, wherein the
first dose and the second dose are separated by 1, 2, 3, 4, 5, 6,
7, 8, 9, or 10 weeks.
[0157] Embodiment 43. The method of embodiment 39, wherein the
first dose and the second dose are separated by 1, 2, 3, 4, 5, or 6
months.
[0158] Embodiment 44. The method of embodiment 39, comprising
administering the pharmaceutical composition monthly or about
monthly.
[0159] Embodiment 45. The method of embodiment 44, comprising
administering the pharmaceutical composition for at least two
months, at least three months, at least four months, at least five
months, or at least six months.
[0160] Embodiment 46. The method of embodiment 39, comprising
administering the pharmaceutical composition weekly or about
weekly.
[0161] Embodiment 47. The method of embodiment 46, comprising
administering the pharmaceutical composition to the animal weekly
or about weekly for less than 2 weeks, less than 3 weeks, less than
4 weeks, less than 5 weeks, less than 6 weeks, less than 8 week,
less than 12 weeks, less than 16 weeks, or less than 20 weeks.
[0162] Embodiment 48. The method of any one of embodiments 27-47,
wherein administering comprises performing a subcutaneous injection
on the animal.
[0163] Embodiment 49. The method of any one of embodiments 27-48,
wherein administering comprises self-administration.
[0164] Embodiment 50. The method of any one of embodiments 27-47,
wherein administering comprises performing an intravenous injection
on the animal.
[0165] Embodiment 51. The method of any one of embodiments 27-50,
comprising administering the pharmaceutical composition at least 1,
at least 2, at least 3, at least 4, at least 5, at least 6, at
least 8, or at least 10 times.
[0166] Embodiment 52. The method of any one of embodiments 27-50,
comprising administering the pharmaceutical composition less than
20 times, less than 15 times, less than 10 times, or less than 5
times.
[0167] Embodiment 53. The method of any one of embodiments 27-52,
wherein the animal has been identified as having a thromboembolic
condition or has been identified as being at risk of having a
thromboembolic condition.
[0168] Embodiment 54. The method of embodiment 53, further
comprising identifying the animal as having the thromboembolic
condition or at risk for having the thromboembolic condition.
[0169] Embodiment 55. The method of any one of embodiments 27-54,
wherein the animal has been identified as having a disease selected
from end stage renal disease (ESRD), chronic kidney disease (CKD),
and coronary artery disease (CAD), or has been identified as being
at risk for a disease selected from ESRD, CKD, and CAD.
[0170] Embodiment 56. The method of embodiment 55, further
comprising identifying the animal as having the disease, or
identifying the animal as being at risk for having the disease.
[0171] Embodiment 57. A method comprising administering a first
dose and a second dose of an oligomeric compound according to the
following formula:
##STR00006##
wherein the first dose and the second dose are separated by 20 to
40 days, and wherein the oligomeric compound is in the form of a
dosage unit consisting or consisting essentially of about 40 mg,
about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg,
about 70 mg, about 75 mg, about 80 mg, about 85 mg, about 90 mg,
about 95 mg, or about 100 mg of the oligomeric compound and about
0.4 ml, about 0.5 ml, about 0.6 ml, about 0.7 ml, about 0.8 ml,
about 0.9 ml, or about 1 ml of a pharmaceutically acceptable
carrier or diluent.
[0172] Embodiment 58. The method of embodiment 57, wherein the
first dose and the second dose are separated by 27 to 32 days, and
wherein the dosage unit consists or consists essentially of 75 mg
to 85 mg of the oligomeric compound and 0.7 ml to 0.9 ml of the
pharmaceutically acceptable carrier or diluent.
[0173] Embodiment 59. The method of embodiment 57, wherein the
dosage unit consists or consists essentially of about 80 mg of the
oligomeric compound and about 0.8 ml of the pharmaceutically
acceptable carrier or diluent.
[0174] Embodiment 60. A method comprising administering a dosage
unit to an animal monthly, wherein the dosage unit consists or
consists essentially of the oligomeric compound of any one of
embodiments 1-5 and a pharmaceutically acceptable carrier or
diluent, and wherein the concentration of the oligomeric compound
is 80 mg/ml to 120 mg/ml.
[0175] Embodiment 61. A method comprising administering a dosage
unit to an animal monthly, wherein the dosage unit consists or
consists essentially of the oligomeric compound of any one of
embodiments 1-5 and a pharmaceutically acceptable carrier or
diluent, and wherein the concentration of the oligomeric compound
is about 100 mg/ml.
[0176] Embodiment 62. The method of any one of embodiments 57 to
61, wherein the pharmaceutically acceptable carrier or diluent is
phosphate buffered saline.
[0177] Embodiment 63. A lyophilized powder comprising the
oligomeric compound of any one of claims 1-5.
[0178] I. Certain Oligonucleotides
[0179] In certain embodiments, provided herein are oligomeric
compounds comprising oligonucleotides, which consist of linked
nucleosides. Oligonucleotides may be unmodified oligonucleotides
(RNA or DNA) or may be modified oligonucleotides. Modified
oligonucleotides comprise at least one modification relative to
unmodified RNA or DNA. That is, modified oligonucleotides comprise
at least one modified nucleoside (comprising a modified sugar
moiety and/or a modified nucleobase) and/or at least one modified
internucleoside linkage.
[0180] A. Certain Modified Nucleosides
[0181] Modified nucleosides comprise a modified sugar moiety or a
modified nucleobase or both a modified sugar moiety and a modified
nucleobase.
[0182] 1. Certain Sugar Moieties
[0183] In certain embodiments, modified sugar moieties are
non-bicyclic modified sugar moieties. In certain embodiments,
modified sugar moieties are bicyclic or tricyclic sugar moieties.
In certain embodiments, modified sugar moieties are sugar
surrogates. Such sugar surrogates may comprise one or more
substitutions corresponding to those of other types of modified
sugar moieties.
[0184] In certain embodiments, modified sugar moieties are
non-bicyclic modified sugar moieties comprising a furanosyl ring
with one or more substituent groups none of which bridges two atoms
of the furanosyl ring to form a bicyclic structure. Such non
bridging substituents may be at any position of the furanosyl,
including but not limited to substituents at the 2', 4', and/or 5'
positions. In certain embodiments one or more non-bridging
substituent of non-bicyclic modified sugar moieties is branched.
Examples of 2'-substituent groups suitable for non-bicyclic
modified sugar moieties include but are not limited to: 2'-F,
2'-OCH.sub.3 ("OMe" or "O-methyl"), and
2'-O(CH.sub.2).sub.2OCH.sub.3 ("MOE"). In certain embodiments,
2'-substituent groups are selected from among: halo, allyl, amino,
azido, SH, CN, OCN, CF.sub.3, OCF.sub.3, O--C.sub.1-C.sub.10
alkoxy, O--C.sub.1-C.sub.10 substituted alkoxy, O--C.sub.1-C.sub.10
alkyl, O--C.sub.1-C.sub.10 substituted alkyl, S-alkyl,
N(R.sub.m)-alkyl, O-alkenyl, S-alkenyl, N(R.sub.m)-alkenyl,
O-alkynyl, S-alkynyl, N(R.sub.m)-alkynyl, O-alkylenyl-O-alkyl,
alkynyl, alkaryl, aralkyl, O-alkaryl, O-aralkyl,
O(CH.sub.2).sub.2SCH.sub.3, O(CH.sub.2).sub.2ON(R.sub.m)(R.sub.n)
or OCH.sub.2C(.dbd.O)--N(R.sub.m)(R.sub.n), where each R.sub.m and
R.sub.n is, independently, H, an amino protecting group, or
substituted or unsubstituted C.sub.1-C.sub.10 alkyl, and the
2'-substituent groups described in Cook et al., U.S. Pat. No.
6,531,584; Cook et al., U.S. Pat. No. 5,859,221; and Cook et al.,
U.S. Pat. No. 6,005,087. Certain embodiments of these
2'-substituent groups can be further substituted with one or more
substituent groups independently selected from among: hydroxyl,
amino, alkoxy, carboxy, benzyl, phenyl, nitro (NO.sub.2), thiol,
thioalkoxy, thioalkyl, halogen, alkyl, aryl, alkenyl and alkynyl.
Examples of 4'-substituent groups suitable for non-bicyclic
modified sugar moieties include but are not limited to alkoxy
(e.g., methoxy), alkyl, and those described in Manoharan et al., WO
2015/106128. Examples of 5'-substituent groups suitable for
non-bicyclic modified sugar moieties include but are not limited
to: 5'-methyl (R or S), 5'-vinyl, and 5'-methoxy. In certain
embodiments, non-bicyclic modified sugar moieties comprise more
than one non-bridging sugar substituent, for example,
2'-F-5'-methyl sugar moieties and the modified sugar moieties and
modified nucleosides described in Migawa et al., WO 2008/101157 and
Rajeev et al., US2013/0203836.).
[0185] In certain embodiments, a 2'-substituted non-bicyclic
modified nucleoside comprises a sugar moiety comprising a
non-bridging 2'-substituent group selected from: F, NH.sub.2,
N.sub.3, OCF.sub.3, OCH.sub.3, O(CH.sub.2).sub.3NH.sub.2,
CH.sub.2CH.dbd.CH.sub.2, OCH.sub.2CH.dbd.CH.sub.2,
OCH.sub.2CH.sub.2OCH.sub.3, O(CH.sub.2).sub.2SCH.sub.3,
O(CH.sub.2).sub.2ON(R.sub.m)(R.sub.n),
O(CH.sub.2).sub.2O(CH.sub.2).sub.2N(CH.sub.3).sub.2, and
N-substituted acetamide (OCH.sub.2C(.dbd.O)--N(R.sub.m)(R.sub.n)),
where each R.sub.m and R.sub.n is, independently, H, an amino
protecting group, or substituted or unsubstituted C.sub.1-C.sub.10
alkyl.
[0186] In certain embodiments, a 2'-substituted nucleoside
non-bicyclic modified nucleoside comprises a sugar moiety
comprising a non-bridging 2'-substituent group selected from: F,
OCF.sub.3, OCH.sub.3, OCH.sub.2CH.sub.2OCH.sub.3,
O(CH.sub.2).sub.2SCH.sub.3, O(CH.sub.2).sub.2ON(CH.sub.3).sub.2,
O(CH.sub.2).sub.2O(CH.sub.2).sub.2N(CH.sub.3).sub.2, and
OCH.sub.2C(.dbd.O)--N(H)CH.sub.3 ("NMA").
[0187] In certain embodiments, a 2'-substituted non-bicyclic
modified nucleoside comprises a sugar moiety comprising a
non-bridging 2'-substituent group selected from: F, OCH.sub.3, and
OCH.sub.2CH.sub.2OCH.sub.3.
[0188] Certain modified sugar moieties comprise a substituent that
bridges two atoms of the furanosyl ring to form a second ring,
resulting in a bicyclic sugar moiety. In certain such embodiments,
the bicyclic sugar moiety comprises a bridge between the 4' and the
2' furanose ring atoms. Examples of such 4' to 2' bridging sugar
substituents include but are not limited to: 4'-CH.sub.2-2',
4'-(CH.sub.2).sub.2-2', 4'-(CH.sub.2).sub.3-2', 4'-CH.sub.2--O-2'
("LNA"), 4'-CH.sub.2-5-2', 4'-(CH.sub.2).sub.2--O-2' ("ENA"),
4'-CH(CH.sub.3)--O-2' (referred to as "constrained ethyl" or
"cEt"), 4'-CH.sub.2--O--CH.sub.2-2', 4'-CH.sub.2--N(R)-2',
4'-CH(CH.sub.2OCH.sub.3)--O-2' ("constrained MOE" or "cMOE") and
analogs thereof (see, e.g., Seth et al., U.S. Pat. No. 7,399,845,
Bhat et al., U.S. Pat. No. 7,569,686, Swayze et al., U.S. Pat. No.
7,741,457, and Swayze et al., U.S. Pat. No. 8,022,193),
4'-C(CH.sub.3)(CH.sub.3)--O-2' and analogs thereof (see, e.g., Seth
et al., U.S. Pat. No. 8,278,283), 4'-CH.sub.2--N(OCH.sub.3)-2' and
analogs thereof (see, e.g., Prakash et al., U.S. Pat. No.
8,278,425), 4'-CH.sub.2--O--N(CH.sub.3)-2' (see, e.g., Allerson et
al., U.S. Pat. No. 7,696,345 and Allerson et al., U.S. Pat. No.
8,124,745), 4'-CH.sub.2--C(H)(CH.sub.3)-2' (see, e.g., Zhou, et
al., J. Org. Chem., 2009, 74, 118-134),
4'-CH.sub.2--C(.dbd.CH.sub.2)-2' and analogs thereof (see e.g.,
Seth et al., U.S. Pat. No. 8,278,426),
4'C(--R.sub.aR.sub.b)--N(R)--O-2',
4'-C(R.sub.aR.sub.b)--O--N(R)-2', 4'-CH.sub.2--O--N(R)-2', and
4'-CH.sub.2--N(R)--O-2', wherein each R, R.sub.a, and R.sub.b is,
independently, H, a protecting group, or C.sub.1-C.sub.12 alkyl
(see, e.g. Imanishi et al., U.S. Pat. No. 7,427,672).
[0189] In certain embodiments, such 4' to 2' bridges independently
comprise from 1 to 4 linked groups independently selected from:
--[C(R.sub.a)(R.sub.b)].sub.n--,
--[C(R.sub.a)(R.sub.b)].sub.n--O--, --C(R.sub.a).dbd.C(R.sub.b)--,
--C(R.sub.a).dbd.N--, C(.dbd.--NR.sub.a)--, --C(.dbd.O)--,
--C(.dbd.S)--, --O--, --Si(R.sub.a).sub.2--, --S(.dbd.O).sub.x--,
and --N(R.sub.a)--;
[0190] wherein:
[0191] x is 0, 1, or 2;
[0192] n is 1, 2, 3, or 4;
[0193] each R.sub.a and R.sub.b is, independently, H, a protecting
group, hydroxyl, C.sub.1-C.sub.12 alkyl, substituted
C.sub.1-C.sub.12 alkyl, C.sub.2-C.sub.12 alkenyl, substituted
C.sub.2-C.sub.12 alkenyl, C.sub.2-C.sub.12 alkynyl, substituted
C.sub.2-C.sub.12 alkynyl, C.sub.5-C.sub.20 aryl, substituted
C.sub.5-C.sub.20 aryl, heterocycle radical, substituted heterocycle
radical, heteroaryl, substituted heteroaryl, C.sub.5-C.sub.7
alicyclic radical, substituted C.sub.5-C.sub.7 alicyclic radical,
halogen, OJ.sub.1, NJ.sub.1J.sub.2, SJ.sub.1, N.sub.3, COOJ.sub.1,
acyl (C(.dbd.O)--H), substituted acyl, CN, sulfonyl
(S(.dbd.O).sub.2-J.sub.1), or sulfoxyl (S(.dbd.O)-J.sub.1); and
[0194] each J.sub.1 and J.sub.2 is, independently, H,
C.sub.1-C.sub.12 alkyl, substituted C.sub.1-C.sub.12 alkyl,
C.sub.2-C.sub.12 alkenyl, substituted C.sub.2-C.sub.12 alkenyl,
C.sub.2-C.sub.12 alkynyl, substituted C.sub.2-C.sub.12 alkynyl,
C.sub.5-C.sub.20 aryl, substituted C.sub.5-C.sub.20 aryl, acyl
(C(.dbd.O)--H), substituted acyl, a heterocycle radical, a
substituted heterocycle radical, C.sub.1-C.sub.12 aminoalkyl,
substituted C.sub.1-C.sub.12 aminoalkyl, or a protecting group.
[0195] Additional bicyclic sugar moieties are known in the art,
see, for example: Freier et al., Nucleic Acids Research, 1997,
25(22), 4429-4443, Albaek et al., J. Org. Chem., 2006, 71,
7731-7740, Singh et al., Chem. Commun., 1998, 4, 455-456; Koshkin
et al., Tetrahedron, 1998, 54, 3607-3630; Kumar et al., Bioorg.
Med. Chem. Lett., 1998, 8, 2219-2222; Singh et al., J. Org. Chem.,
1998, 63, 10035-10039; Srivastava et al., J. Am. Chem. Soc., 20017,
129, 8362-8379; Wengel et a., U.S. Pat. No. 7,053,207; Imanishi et
al., U.S. Pat. No. 6,268,490; Imanishi et al. U.S. Pat. No.
6,770,748; Imanishi et al., U.S. Pat. No. RE44,779; Wengel et al.,
U.S. Pat. No. 6,794,499; Wengel et al., U.S. Pat. No. 6,670,461;
Wengel et al., U.S. Pat. No. 7,034,133; Wengel et al., U.S. Pat.
No. 8,080,644; Wengel et al., U.S. Pat. No. 8,034,909; Wengel et
al., U.S. Pat. No. 8,153,365; Wengel et al., U.S. Pat. No.
7,572,582; and Ramasamy et al., U.S. Pat. No. 6,525,191; Torsten et
al., WO 2004/106356; Wengel et al., WO 1999/014226; Seth et al., WO
2007/134181; Seth et al., U.S. Pat. No. 7,547,684; Seth et al.,
U.S. Pat. No. 7,666,854; Seth et al., U.S. Pat. No. 8,088,746; Seth
et al., U.S. Pat. No. 7,750,131; Seth et al., U.S. Pat. No.
8,030,467; Seth et al., U.S. Pat. No. 8,268,980; Seth et al., U.S.
Pat. No. 8,546,556; Seth et al., U.S. Pat. No. 8,530,640; Migawa et
al., U.S. Pat. No. 9,012,421; Seth et al., U.S. Pat. No. 8,501,805;
and U.S. Patent Publication Nos. Allerson et al., US2008/0039618
and Migawa et al., US2015/0191727.
[0196] In certain embodiments, bicyclic sugar moieties and
nucleosides incorporating such bicyclic sugar moieties are further
defined by isomeric configuration. For example, an LNA nucleoside
(described herein) may be in the .alpha.-L configuration or in the
.beta.-D configuration.
##STR00007##
.alpha.-L-methyleneoxy (4'-CH.sub.2--O-2') or .alpha.-L-LNA
bicyclic nucleosides have been incorporated into oligonucleotides
that showed antisense activity (Frieden et al., Nucleic Acids
Research, 2003, 21, 6365-6372). Herein, general descriptions of
bicyclic nucleosides include both isomeric configurations. When the
positions of specific bicyclic nucleosides (e.g., LNA or cEt) are
identified in exemplified embodiments herein, they are in the
.beta.-D configuration, unless otherwise specified.
[0197] In certain embodiments, modified sugar moieties comprise one
or more non-bridging sugar substituent and one or more bridging
sugar substituent (e.g., 5'-substituted and 4'-2' bridged
sugars).
[0198] In certain embodiments, modified sugar moieties are sugar
surrogates. In certain such embodiments, the oxygen atom of the
sugar moiety is replaced, e.g., with a sulfur, carbon or nitrogen
atom. In certain such embodiments, such modified sugar moieties
also comprise bridging and/or non-bridging substituents as
described herein. For example, certain sugar surrogates comprise a
4'-sulfur atom and a substitution at the 2'-position (see, e.g.,
Bhat et al., U.S. Pat. No. 7,875,733 and Bhat et al., U.S. Pat. No.
7,939,677) and/or the 5' position.
[0199] In certain embodiments, sugar surrogates comprise rings
having other than 5 atoms. For example, in certain embodiments, a
sugar surrogate comprises a six-membered tetrahydropyran ("THP").
Such tetrahydropyrans may be further modified or substituted.
Nucleosides comprising such modified tetrahydropyrans include but
are not limited to hexitol nucleic acid ("HNA"), anitol nucleic
acid ("ANA"), manitol nucleic acid ("MNA") (see, e.g., Leumann, C
J. Bioorg. & Med. Chem. 2002, 10, 841-854), fluoro HNA:
##STR00008##
("F-HNA", see e.g., Swayze et al., U.S. Pat. No. 8,088,904; Swayze
et al., U.S. Pat. No. 8,440,803; Swayze et al., U.S. Pat. No.
8,796,437; and Swayze et al., U.S. Pat. No. 9,005,906; F-HNA can
also be referred to as a F-THP or 3'-fluoro tetrahydropyran), and
nucleosides comprising additional modified THP compounds having the
formula:
##STR00009##
wherein, independently, for each of said modified THP
nucleoside:
[0200] Bx is a nucleobase moiety;
[0201] T.sub.3 and T.sub.4 are each, independently, an
internucleoside linking group linking the modified THP nucleoside
to the remainder of an oligonucleotide or one of T.sub.3 and
T.sub.4 is an internucleoside linking group linking the modified
THP nucleoside to the remainder of an oligonucleotide and the other
of T.sub.3 and T.sub.4 is H, a hydroxyl protecting group, a linked
conjugate group, or a 5' or 3'-terminal group; q.sub.1, q.sub.2,
q.sub.3, q.sub.4, q.sub.5, q.sub.6 and q.sub.7 are each,
independently, H, C.sub.1-C.sub.6 alkyl, substituted
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, substituted
C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, or substituted
C.sub.2-C.sub.6 alkynyl; and
[0202] each of R.sub.1 and R.sub.2 is independently selected from
among: hydrogen, halogen, substituted or unsubstituted alkoxy,
NJ.sub.1J.sub.2, SJ.sub.1, N.sub.3, OC(.dbd.X)J.sub.1,
OC(.dbd.X)NJ.sub.1J.sub.2, NJ.sub.3C(.dbd.X)NJ.sub.1J.sub.2, and
CN, wherein X is O, S or NJ.sub.1, and each J.sub.1, J.sub.2, and
J.sub.3 is, independently, H or C.sub.1-C.sub.6 alkyl.
[0203] In certain embodiments, modified THP nucleosides are
provided wherein q.sub.1, q.sub.2, q.sub.3, q.sub.4, q.sub.5,
q.sub.6 and q.sub.7 are each H. In certain embodiments, at least
one of q.sub.1, q.sub.2, q.sub.3, q.sub.4, q.sub.5, q.sub.6 and
q.sub.7 is other than H. In certain embodiments, at least one of
q.sub.1, q.sub.2, q.sub.3, q.sub.4, q.sub.5, q.sub.6 and q.sub.7 is
methyl. In certain embodiments, modified THP nucleosides are
provided wherein one of R.sub.1 and R.sub.2 is F. In certain
embodiments, R.sub.1 is F and R.sub.2 is H, in certain embodiments,
R.sub.1 is methoxy and R.sub.2 is H, and in certain embodiments,
R.sub.1 is methoxyethoxy and R.sub.2 is H.
[0204] In certain embodiments, sugar surrogates comprise rings
having more than 5 atoms and more than one heteroatom. For example,
nucleosides comprising morpholino sugar moieties and their use in
oligonucleotides have been reported (see, e.g., Braasch et al.,
Biochemistry, 2002, 41, 4503-4510 and Summerton et al., U.S. Pat.
No. 5,698,685; Summerton et al., U.S. Pat. No. 5,166,315; Summerton
et al., U.S. Pat. No. 5,185,444; and Summerton et al., U.S. Pat.
No. 5,034,506). As used here, the term "morpholino" means a sugar
surrogate having the following structure:
##STR00010##
In certain embodiments, morpholinos may be modified, for example by
adding or altering various substituent groups from the above
morpholino structure. Such sugar surrogates are referred to herein
as "modifed morpholinos."
[0205] In certain embodiments, sugar surrogates comprise acyclic
moieites. Examples of nucleosides and oligonucleotides comprising
such acyclic sugar surrogates include but are not limited to:
peptide nucleic acid ("PNA"), acyclic butyl nucleic acid (see,
e.g., Kumar et al., Org. Biomol. Chem., 2013, 11, 5853-5865), and
nucleosides and oligonucleotides described in Manoharan et al.,
WO2011/133876.
[0206] Many other bicyclic and tricyclic sugar and sugar surrogate
ring systems are known in the art that can be used in modified
nucleosides (see for example review article: Leumann, Bioorg. Med.
Chem., 2002, 10, 841-854).
[0207] 2. Certain Modified Nucleobases
[0208] In certain embodiments, modified oligonucleotides comprise
one or more nucleosides comprising an unmodified nucleobase. In
certain embodiments, modified oligonucleotides comprise one or more
nucleoside comprising a modified nucleobase. In certain
embodiments, modified oligonucleotides comprise one or more
nucleoside that does not comprise a nucleobase, referred to as an
abasic nucleoside.
[0209] In certain embodiments, modified nucleobases are selected
from: 5-substituted pyrimidines, 6-azapyrimidines, alkyl or alkynyl
substituted pyrimidines, alkyl substituted purines, and N-2, N-6
and 0-6 substituted purines. In certain embodiments, modified
nucleobases are selected from: 2-aminopropyladenine,
5-hydroxymethyl cytosine, xanthine, hypoxanthine, 2-aminoadenine,
6-N-methylguanine, 6-N-methyladenine, 2-propyladenine,
2-thiouracil, 2-thiothymine and 2-thiocytosine, 5-propynyl
(--C.ident.C--CH.sub.3) uracil, 5-propynylcytosine, 6-azouracil,
6-azocytosine, 6-azothymine, 5-ribosyluracil (pseudouracil),
4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl,
8-aza and other 8-substituted purines, 5-halo, particularly
5-bromo, 5-trifluoromethyl, 5-halouracil, and 5-halocytosine,
7-methylguanine, 7-methyladenine, 2-F-adenine, 2-aminoadenine,
7-deazaguanine, 7-deazaadenine, 3-deazaguanine, 3-deazaadenine,
6-N-benzoyladenine, 2-N-isobutyrylguanine, 4-N-benzoylcytosine,
4-N-benzoyluracil, 5-methyl 4-N-benzoylcytosine, 5-methyl
4-N-benzoyluracil, universal bases, hydrophobic bases, promiscuous
bases, size-expanded bases, and fluorinated bases. Further modified
nucleobases include tricyclic pyrimidines, such as
1,3-diazaphenoxazine-2-one, 1,3-diazaphenothiazine-2-one and
9-(2-aminoethoxy)-1,3-diazaphenoxazine-2-one (G-clamp). Modified
nucleobases may also include those in which the purine or
pyrimidine base is replaced with other heterocycles, for example
7-deaza-adenine, 7-deazaguanosine, 2-aminopyridine and 2-pyridone.
Further nucleobases include those disclosed in Merigan et al., U.S.
Pat. No. 3,687,808, those disclosed in The Concise Encyclopedia Of
Polymer Science And Engineering, Kroschwitz, J. I., Ed., John Wiley
& Sons, 1990, 858-859; Englisch et al., Angewandte Chemie,
International Edition, 1991, 30, 613; Sanghvi, Y. S., Chapter 15,
Antisense Research and Applications, Crooke, S. T. and Lebleu, B.,
Eds., CRC Press, 1993, 273-288; and those disclosed in Chapters 6
and 15, Antisense Drug Technology, Crooke S. T., Ed., CRC Press,
2008, 163-166 and 442-443.
[0210] Publications that teach the preparation of certain of the
above noted modified nucleobases as well as other modified
nucleobases include without limitation, Manoharan et al.,
US2003/0158403; Manoharan et al., US2003/0175906; Dinh et al., U.S.
Pat. No. 4,845,205; Spielvogel et al., U.S. Pat. No. 5,130,302;
Rogers et al., U.S. Pat. No. 5,134,066; Bischofberger et al., U.S.
Pat. No. 5,175,273; Urdea et al., U.S. Pat. No. 5,367,066; Benner
et al., U.S. Pat. No. 5,432,272; Matteucci et al., U.S. Pat. No.
5,434,257; Gmeiner et al., U.S. Pat. No. 5,457,187; Cook et al.,
U.S. Pat. No. 5,459,255; Froehler et al., U.S. Pat. No. 5,484,908;
Matteucci et al., U.S. Pat. No. 5,502,177; Hawkins et al., U.S.
Pat. No. 5,525,711; Haralambidis et al., U.S. Pat. No. 5,552,540;
Cook et al., U.S. Pat. No. 5,587,469; Froehler et al., U.S. Pat.
No. 5,594,121; Switzer et al., U.S. Pat. No. 5,596,091; Cook et
al., U.S. Pat. No. 5,614,617; Froehler et al., U.S. Pat. No.
5,645,985; Cook et al., U.S. Pat. No. 5,681,941; Cook et al., U.S.
Pat. No. 5,811,534; Cook et al., U.S. Pat. No. 5,750,692; Cook et
al., U.S. Pat. No. 5,948,903; Cook et al., U.S. Pat. No. 5,587,470;
Cook et al., U.S. Pat. No. 5,457,191; Matteucci et al., U.S. Pat.
No. 5,763,588; Froehler et al., U.S. Pat. No. 5,830,653; Cook et
al., U.S. Pat. No. 5,808,027; Cook et al., U.S. Pat. No. 6,166,199;
and Matteucci et al., U.S. Pat. No. 6,005,096.
[0211] 3. Certain Modified Internucleoside Linkages
[0212] In certain embodiments, nucleosides of modified
oligonucleotides may be linked together using any internucleoside
linkage. The two main classes of internucleoside linking groups are
defined by the presence or absence of a phosphorus atom.
Representative phosphorus-containing internucleoside linkages
include but are not limited to phosphodiesters ("P.dbd.O") (also
referred to as unmodified or naturally occurring linkages or
phosphate linkages), phosphotriesters, methylphosphonates,
phosphoramidates, and phosphorothioates ("P.dbd.S"), and
phosphorodithioates ("HS--P.dbd.S"). Representative non-phosphorus
containing internucleoside linking groups include but are not
limited to methylenemethylimino
(--CH.sub.2--N(CH.sub.3)--O--CH.sub.2--), thiodiester,
thionocarbamate (--O--C(.dbd.O)(NH)--S--); siloxane
(--O--SiH.sub.2--O--); and N,N'-dimethylhydrazine
(--CH.sub.2--N(CH.sub.3)--N(CH.sub.3)--). Modified internucleoside
linkages, compared to naturally occurring phosphodiester linkages,
can be used to alter, typically increase, nuclease resistance of
the oligonucleotide. In certain embodiments, internucleoside
linkages having a chiral atom can be prepared as a racemic mixture,
or as separate enantiomers. Methods of preparation of
phosphorous-containing and non-phosphorous-containing
internucleoside linkages are well known to those skilled in the
art.
[0213] Representative internucleoside linkages having a chiral
center include but are not limited to alkylphosphonates and
phosphorothioates. Modified oligonucleotides comprising
internucleoside linkages having a chiral center can be prepared as
populations of modified oligonucleotides comprising stereorandom
internucleoside linkages, or as populations of modified
oligonucleotides comprising phosphorothioate linkages in particular
stereochemical configurations. In certain embodiments, populations
of modified oligonucleotides comprise phosphorothioate
internucleoside linkages wherein all of the phosphorothioate
internucleoside linkages are stereorandom. Such modified
oligonucleotides can be generated using synthetic methods that
result in random selection of the stereochemical configuration of
each phosphorothioate linkage. Nonetheless, as is well understood
by those of skill in the art, each individual phosphorothioate of
each individual oligonucleotide molecule has a defined
stereoconfiguration. In certain embodiments, populations of
modified oligonucleotides are enriched for modified
oligonucleotides comprising one or more particular phosphorothioate
internucleoside linkages in a particular, independently selected
stereochemical configuration. In certain embodiments, the
particular configuration of the particular phosphorothioate linkage
is present in at least 65% of the molecules in the population. In
certain embodiments, the particular configuration of the particular
phosphorothioate linkage is present in at least 70% of the
molecules in the population. In certain embodiments, the particular
configuration of the particular phosphorothioate linkage is present
in at least 80% of the molecules in the population. In certain
embodiments, the particular configuration of the particular
phosphorothioate linkage is present in at least 90% of the
molecules in the population. In certain embodiments, the particular
configuration of the particular phosphorothioate linkage is present
in at least 99% of the molecules in the population. Such chirally
enriched populations of modified oligonucleotides can be generated
using synthetic methods known in the art, e.g., methods described
in Oka et al., JACS 125, 8307 (2003), Wan et al. Nuc. Acid. Res.
42, 13456 (2014), and WO 2017/015555. In certain embodiments, a
population of modified oligonucleotides is enriched for modified
oligonucleotides having at least one indicated phosphorothioate in
the (Sp) configuration. In certain embodiments, a population of
modified oligonucleotides is enriched for modified oligonucleotides
having at least one phosphorothioate in the (Rp) configuration. In
certain embodiments, modified oligonucleotides comprising (Rp)
and/or (Sp) phosphorothioates comprise one or more of the following
formulas, respectively, wherein "B" indicates a nucleobase:
##STR00011##
Unless otherwise indicated, chiral internucleoside linkages of
modified oligonucleotides described herein can be stereorandom or
in a particular stereochemical configuration.
[0214] Neutral internucleoside linkages include, without
limitation, phosphotriesters, methylphosphonates, MMI
(3'-CH.sub.2--N(CH.sub.3)--O-5'), amide-3
(3'-CH.sub.2--C(.dbd.O)--N(H)-5'), amide-4
(3'-CH.sub.2--N(H)--C(.dbd.O)-5'), formacetal
(3'-O--CH.sub.2--O-5'), methoxypropyl, and thioformacetal
(3'-S--CH.sub.2--O-5'). Further neutral internucleoside linkages
include nonionic linkages comprising siloxane (dialkylsiloxane),
carboxylate ester, carboxamide, sulfide, sulfonate ester and amides
(See for example: Carbohydrate Modifications in Antisense Research;
Y. S. Sanghvi and P. D. Cook, Eds., ACS Symposium Series 580;
Chapters 3 and 4, 40-65). Further neutral internucleoside linkages
include nonionic linkages comprising mixed N, O, S and CH.sub.2
component parts.
[0215] B. Certain Motifs
[0216] In certain embodiments, modified oligonucleotides comprise
one or more modified nucleosides comprising a modified sugar
moiety. In certain embodiments, modified oligonucleotides comprise
one or more modified nucleosides comprising a modified nucleobase.
In certain embodiments, modified oligonucleotides comprise one or
more modified internucleoside linkage. In such embodiments, the
modified, unmodified, and differently modified sugar moieties,
nucleobases, and/or internucleoside linkages of a modified
oligonucleotide define a pattern or motif. In certain embodiments,
the patterns of sugar moieties, nucleobases, and internucleoside
linkages are each independent of one another. Thus, a modified
oligonucleotide may be described by its sugar motif, nucleobase
motif and/or internucleoside linkage motif (as used herein,
nucleobase motif describes the modifications to the nucleobases
independent of the sequence of nucleobases).
[0217] 1. Certain Sugar Motifs
[0218] In certain embodiments, oligonucleotides comprise one or
more type of modified sugar and/or unmodified sugar moiety arranged
along the oligonucleotide or region thereof in a defined pattern or
sugar motif. In certain instances, such sugar motifs include but
are not limited to any of the sugar modifications discussed
herein.
[0219] In certain embodiments, modified oligonucleotides comprise
or consist of a region having a gapmer motif, which is defined by
two external regions or "wings" and a central or internal region or
"gap." The three regions of a gapmer motif (the 5'-wing, the gap,
and the 3'-wing) form a contiguous sequence of nucleosides wherein
at least some of the sugar moieties of the nucleosides of each of
the wings differ from at least some of the sugar moieties of the
nucleosides of the gap. Specifically, at least the sugar moieties
of the nucleosides of each wing that are closest to the gap (the
3'-most nucleoside of the 5'-wing and the 5'-most nucleoside of the
3'-wing) differ from the sugar moiety of the neighboring gap
nucleosides, thus defining the boundary between the wings and the
gap (i.e., the wing/gap junction). In certain embodiments, the
sugar moieties within the gap are the same as one another. In
certain embodiments, the gap includes one or more nucleoside having
a sugar moiety that differs from the sugar moiety of one or more
other nucleosides of the gap. In certain embodiments, the sugar
motifs of the two wings are the same as one another (symmetric
gapmer). In certain embodiments, the sugar motif of the 5'-wing
differs from the sugar motif of the 3'-wing (asymmetric
gapmer).
[0220] In certain embodiments, the wings of a gapmer comprise 1-5
nucleosides. In certain embodiments, each nucleoside of each wing
of a gapmer is a modified nucleoside. In certain embodiments, at
least one nucleoside of each wing of a gapmer is a modified
nucleoside. In certain embodiments, at least two nucleosides of
each wing of a gapmer are modified nucleosides. In certain
embodiments, at least three nucleosides of each wing of a gapmer
are modified nucleosides. In certain embodiments, at least four
nucleosides of each wing of a gapmer are modified nucleosides.
[0221] In certain embodiments, the gap of a gapmer comprises 7-12
nucleosides. In certain embodiments, each nucleoside of the gap of
a gapmer is an unmodified 2'-deoxy nucleoside.
[0222] In certain embodiments, the gapmer is a deoxy gapmer. In
embodiments, the nucleosides on the gap side of each wing/gap
junction are unmodified 2'-deoxy nucleosides and the nucleosides on
the wing sides of each wing/gap junction are modified nucleosides.
In certain embodiments, each nucleoside of the gap is an unmodified
2'-deoxy nucleoside. In certain embodiments, each nucleoside of
each wing of a gapmer is a modified nucleoside.
[0223] In certain embodiments, modified oligonucleotides comprise
or consist of a region having a fully modified sugar motif. In such
embodiments, each nucleoside of the fully modified region of the
modified oligonucleotide comprises a modified sugar moiety. In
certain embodiments, each nucleoside of the entire modified
oligonucleotide comprises a modified sugar moiety. In certain
embodiments, modified oligonucleotides comprise or consist of a
region having a fully modified sugar motif, wherein each nucleoside
within the fully modified region comprises the same modified sugar
moiety, referred to herein as a uniformly modified sugar motif. In
certain embodiments, a fully modified oligonucleotide is a
uniformly modified oligonucleotide. In certain embodiments, each
nucleoside of a uniformly modified comprises the same
2'-modification.
[0224] Herein, the lengths (number of nucleosides) of the three
regions of a gapmer may be provided using the notation [# of
nucleosides in the 5'-wing]--[# of nucleosides in the gap]--[# of
nucleosides in the 3'-wing]. Thus, a 5-10-5 gapmer consists of 5
linked nucleosides in each wing and 10 linked nucleosides in the
gap. Where such nomenclature is followed by a specific
modification, that modification is the modification in each sugar
moiety of each wing and the gap nucleosides comprise unmodified
deoxynucleoside sugars. Thus, a 5-10-5 MOE gapmer consists of 5
linked MOE modified nucleosides in the 5'-wing, 10 linked
deoxynucleosides in the gap, and 5 linked MOE nucleosides in the
3'-wing.
[0225] In certain embodiments, modified oligonucleotides are 5-10-5
MOE gapmers. In certain embodiments, modified oligonucleotides are
3-10-3 BNA gapmers. In certain embodiments, modified
oligonucleotides are 3-10-3 cEt gapmers. In certain embodiments,
modified oligonucleotides are 3-10-3 LNA gapmers.
[0226] 2. Certain Nucleobase Motifs
[0227] In certain embodiments, oligonucleotides comprise modified
and/or unmodified nucleobases arranged along the oligonucleotide or
region thereof in a defined pattern or motif. In certain
embodiments, each nucleobase is modified. In certain embodiments,
none of the nucleobases are modified. In certain embodiments, each
purine or each pyrimidine is modified. In certain embodiments, each
adenine is modified. In certain embodiments, each guanine is
modified. In certain embodiments, each thymine is modified. In
certain embodiments, each uracil is modified. In certain
embodiments, each cytosine is modified. In certain embodiments,
some or all of the cytosine nucleobases in a modified
oligonucleotide are 5-methyl cytosines. In certain embodiments, all
of the cytosine nucleobases are 5-methyl cytosines and all of the
other nucleobases of the modified oligonucleotide are unmodified
nucleobases.
[0228] In certain embodiments, modified oligonucleotides comprise a
block of modified nucleobases. In certain such embodiments, the
block is at the 3'-end of the oligonucleotide. In certain
embodiments the block is within 3 nucleosides of the 3'-end of the
oligonucleotide. In certain embodiments, the block is at the 5'-end
of the oligonucleotide. In certain embodiments the block is within
3 nucleosides of the 5'-end of the oligonucleotide.
[0229] In certain embodiments, oligonucleotides having a gapmer
motif comprise a nucleoside comprising a modified nucleobase. In
certain such embodiments, one nucleoside comprising a modified
nucleobase is in the central gap of an oligonucleotide having a
gapmer motif In certain such embodiments, the sugar moiety of said
nucleoside is a 2'-deoxyribosyl moiety. In certain embodiments, the
modified nucleobase is selected from: a 2-thiopyrimidine and a
5-propynepyrimidine.
[0230] 3. Certain Internucleoside Linkage Motifs
[0231] In certain embodiments, oligonucleotides comprise modified
and/or unmodified internucleoside linkages arranged along the
oligonucleotide or region thereof in a defined pattern or motif. In
certain embodiments, each internucleoside linking group is a
phosphodiester internucleoside linkage (P.dbd.O). In certain
embodiments, each internucleoside linking group of a modified
oligonucleotide is a phosphorothioate internucleoside linkage
(P.dbd.S). In certain embodiments, each internucleoside linkage of
a modified oligonucleotide is independently selected from a
phosphorothioate internucleoside linkage and phosphodiester
internucleoside linkage. In certain embodiments, each
phosphorothioate internucleoside linkage is independently selected
from a stereorandom phosphorothioate, a (Sp) phosphorothioate, and
a (Rp) phosphorothioate. In certain embodiments, the sugar motif of
a modified oligonucleotide is a gapmer and the internucleoside
linkages within the gap are all modified. In certain such
embodiments, some or all of the internucleoside linkages in the
wings are unmodified phosphodiester internucleoside linkages. In
certain embodiments, the terminal internucleoside linkages are
modified. In certain embodiments, the sugar motif of a modified
oligonucleotide is a gapmer, and the internucleoside linkage motif
comprises at least one phosphodiester internucleoside linkage in at
least one wing, wherein the at least one phosphodiester linkage is
not a terminal internucleoside linkage, and the remaining
internucleoside linkages are phosphorothioate internucleoside
linkages. In certain such embodiments, all of the phosphorothioate
linkages are stereorandom. In certain embodiments, all of the
phosphorothioate linkages in the wings are (Sp) phosphorothioates,
and the gap comprises at least one Sp, Sp, Rp motif. In certain
embodiments, populations of modified oligonucleotides are enriched
for modified oligonucleotides comprising such internucleoside
linkage motifs.
[0232] C. Certain Lengths
[0233] It is possible to increase or decrease the length of an
oligonucleotide without eliminating activity. For example, in Woolf
et al. (Proc. Natl. Acad. Sci. USA 89:7305-7309, 1992), a series of
oligonucleotides 13-25 nucleobases in length were tested for their
ability to induce cleavage of a target RNA in an oocyte injection
model. Oligonucleotides 25 nucleobases in length with 8 or 11
mismatch bases near the ends of the oligonucleotides were able to
direct specific cleavage of the target RNA, albeit to a lesser
extent than the oligonucleotides that contained no mismatches.
Similarly, target specific cleavage was achieved using 13
nucleobase oligonucleotides, including those with 1 or 3
mismatches.
[0234] In certain embodiments, oligonucleotides (including modified
oligonucleotides) can have any of a variety of ranges of lengths.
In certain embodiments, oligonucleotides consist of X to Y linked
nucleosides, where X represents the fewest number of nucleosides in
the range and Y represents the largest number nucleosides in the
range. In certain such embodiments, X and Y are each independently
selected from 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21,
22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38,
39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, and 50; provided that
X.ltoreq.Y. For example, in certain embodiments, oligonucleotides
consist of 12 to 13, 12 to 14, 12 to 15, 12 to 16, 12 to 17, 12 to
18, 12 to 19, 12 to 20, 12 to 21, 12 to 22, 12 to 23, 12 to 24, 12
to 25, 12 to 26, 12 to 27, 12 to 28, 12 to 29, 12 to 30, 13 to 14,
13 to 15, 13 to 16, 13 to 17, 13 to 18, 13 to 19, 13 to 20, 13 to
21, 13 to 22, 13 to 23, 13 to 24, 13 to 25, 13 to 26, 13 to 27, 13
to 28, 13 to 29, 13 to 30, 14 to 15, 14 to 16, 14 to 17, 14 to 18,
14 to 19, 14 to 20, 14 to 21, 14 to 22, 14 to 23, 14 to 24, 14 to
25, 14 to 26, 14 to 27, 14 to 28, 14 to 29, 14 to 30, 15 to 16, 15
to 17, 15 to 18, 15 to 19, 15 to 20, 15 to 21, 15 to 22, 15 to 23,
15 to 24, 15 to 25, 15 to 26, 15 to 27, 15 to 28, 15 to 29, 15 to
30, 16 to 17, 16 to 18, 16 to 19, 16 to 20, 16 to 21, 16 to 22, 16
to 23, 16 to 24, 16 to 25, 16 to 26, 16 to 27, 16 to 28, 16 to 29,
16 to 30, 17 to 18, 17 to 19, 17 to 20, 17 to 21, 17 to 22, 17 to
23, 17 to 24, 17 to 25, 17 to 26, 17 to 27, 17 to 28, 17 to 29, 17
to 30, 18 to 19, 18 to 20, 18 to 21, 18 to 22, 18 to 23, 18 to 24,
18 to 25, 18 to 26, 18 to 27, 18 to 28, 18 to 29, 18 to 30, 19 to
20, 19 to 21, 19 to 22, 19 to 23, 19 to 24, 19 to 25, 19 to 26, 19
to 29, 19 to 28, 19 to 29, 19 to 30, 20 to 21, 20 to 22, 20 to 23,
20 to 24, 20 to 25, 20 to 26, 20 to 27, 20 to 28, 20 to 29, 20 to
30, 21 to 22, 21 to 23, 21 to 24, 21 to 25, 21 to 26, 21 to 27, 21
to 28, 21 to 29, 21 to 30, 22 to 23, 22 to 24, 22 to 25, 22 to 26,
22 to 27, 22 to 28, 22 to 29, 22 to 30, 23 to 24, 23 to 25, 23 to
26, 23 to 27, 23 to 28, 23 to 29, 23 to 30, 24 to 25, 24 to 26, 24
to 27, 24 to 28, 24 to 29, 24 to 30, 25 to 26, 25 to 27, 25 to 28,
25 to 29, 25 to 30, 26 to 27, 26 to 28, 26 to 29, 26 to 30, 27 to
28, 27 to 29, 27 to 30, 28 to 29, 28 to 30, or 29 to 30 linked
nucleosides
[0235] D. Certain Modified Oligonucleotides
[0236] In certain embodiments, the above modifications (sugar,
nucleobase, internucleoside linkage) are incorporated into a
modified oligonucleotide. In certain embodiments, modified
oligonucleotides are characterized by their modification motifs and
overall lengths. In certain embodiments, such parameters are each
independent of one another. Thus, unless otherwise indicated, each
internucleoside linkage of an oligonucleotide having a gapmer sugar
motif may be modified or unmodified and may or may not follow the
gapmer modification pattern of the sugar modifications. For
example, the internucleoside linkages within the wing regions of a
sugar gapmer may be the same or different from one another and may
be the same or different from the internucleoside linkages of the
gap region of the sugar motif. Likewise, such sugar gapmer
oligonucleotides may comprise one or more modified nucleobase
independent of the gapmer pattern of the sugar modifications.
Unless otherwise indicated, all modifications are independent of
nucleobase sequence.
[0237] E. Certain Populations of Modified Oligonucleotides
[0238] Populations of modified oligonucleotides in which all of the
modified oligonucleotides of the population have the same molecular
formula can be stereorandom populations or chirally enriched
populations. All of the chiral centers of all of the modified
oligonucleotides are stereorandom in a stereorandom population. In
a chirally enriched population, at least one particular chiral
center is not stereorandom in the modified oligonucleotides of the
population. In certain embodiments, the modified oligonucleotides
of a chirally enriched population are enriched for .beta.-D ribosyl
sugar moieties, and all of the phosphorothioate internucleoside
linkages are stereorandom. In certain embodiments, the modified
oligonucleotides of a chirally enriched population are enriched for
both .beta.-D ribosyl sugar moieties and at least one, particular
phosphorothioate internucleoside linkage in a particular
stereochemical configuration.
[0239] F. Nucleobase Sequence
[0240] In certain embodiments, oligonucleotides (unmodified or
modified oligonucleotides) are further described by their
nucleobase sequence. In certain embodiments oligonucleotides have a
nucleobase sequence that is complementary to a second
oligonucleotide or an identified reference nucleic acid, such as a
target nucleic acid. In certain such embodiments, a region of an
oligonucleotide has a nucleobase sequence that is complementary to
a second oligonucleotide or an identified reference nucleic acid,
such as a target nucleic acid. In certain embodiments, the
nucleobase sequence of a region or entire length of an
oligonucleotide is at least 50%, at least 60%, at least 70%, at
least 80%, at least 85%, at least 90%, at least 95%, or 100%
complementary to the second oligonucleotide or nucleic acid, such
as a target nucleic acid.
[0241] II. Certain Oligomeric Compounds
[0242] In certain embodiments, provided herein are oligomeric
compounds, which consist of an oligonucleotide (modified or
unmodified) and optionally one or more conjugate groups and/or
terminal groups. Conjugate groups consist of one or more conjugate
moieties and a conjugate linker which links the conjugate moiety to
the oligonucleotide. Conjugate groups may be attached to either or
both ends of an oligonucleotide and/or at any internal position. In
certain embodiments, conjugate groups are attached to the
2'-position of a nucleoside of a modified oligonucleotide. In
certain embodiments, conjugate groups are attached to the 3' and/or
5' end of oligonucleotides. In certain embodiments, conjugate
groups that are attached to either or both ends of an
oligonucleotide are terminal groups. In certain such embodiments,
conjugate groups (or terminal groups) are attached at the 3'-end of
oligonucleotides. In certain embodiments, conjugate groups are
attached near the 3'-end of oligonucleotides. In certain
embodiments, conjugate groups (or terminal groups) are attached at
the 5'-end of oligonucleotides. In certain embodiments, conjugate
groups are attached near the 5'-end of oligonucleotides.
[0243] Examples of terminal groups include but are not limited to
conjugate groups, capping groups, phosphate moieties, protecting
groups, modified or unmodified nucleosides, and two or more
nucleosides that are independently modified or unmodified.
[0244] A. Certain Conjugate Groups
[0245] In certain embodiments, oligonucleotides are covalently
attached to one or more conjugate groups. In certain embodiments,
conjugate groups modify one or more properties of the attached
oligonucleotide, including but not limited to pharmacodynamics,
pharmacokinetics, stability, binding, absorption, tissue
distribution, cellular distribution, cellular uptake, charge and
clearance. In certain embodiments, conjugate groups impart a new
property on the attached oligonucleotide, e.g., fluorophores or
reporter groups that enable detection of the oligonucleotide.
Certain conjugate groups and conjugate moieties have been described
previously, for example: cholesterol moiety (Letsinger et al.,
Proc. Natl. Acad. Sci. USA, 1989, 86, 6553-6556), cholic acid
(Manoharan et al., Bioorg. Med. Chem. Lett., 1994, 4, 1053-1060), a
thioether, e.g., hexyl-S-tritylthiol (Manoharan et al., Ann. N.Y.
Acad. Sci., 1992, 660, 306-309; Manoharan et al., Bioorg. Med.
Chem. Lett., 1993, 3, 2765-2770), a thiocholesterol (Oberhauser et
al., Nucl. Acids Res., 1992, 20, 533-538), an aliphatic chain,
e.g., do-decan-diol or undecyl residues (Saison-Behmoaras et al.,
EMBO 1, 1991, 10, 1111-1118; Kabanov et al., FEBS Lett., 1990, 259,
327-330; Svinarchuk et al., Biochimie, 1993, 75, 49-54), a
phospholipid, e.g., di-hexadecyl-rac-glycerol or triethyl-ammonium
1,2-di-O-hexadecyl-rac-glycero-3-H-phosphonate (Manoharan et al.,
Tetrahedron Lett., 1995, 36, 3651-3654; Shea et al., Nucl. Acids
Res., 1990, 18, 3777-3783), a polyamine or a polyethylene glycol
chain (Manoharan et al., Nucleosides & Nucleotides, 1995, 14,
969-973), or adamantane acetic acid a palmityl moiety (Mishra et
al., Biochim. Biophys. Acta, 1995, 1264, 229-237), an
octadecylamine or hexylamino-carbonyl-oxycholesterol moiety (Crooke
et al., J. Pharmacol. Exp. Ther., 1996, 277, 923-937), a tocopherol
group (Nishina et al., Molecular Therapy Nucleic Acids, 2015, 4,
e220; and Nishina et al., Molecular Therapy, 2008, 16, 734-740), or
a GalNAc cluster (e.g., WO2014/179620).
[0246] 1. Conjugate Moieties
[0247] Conjugate moieties include, without limitation,
intercalators, reporter molecules, polyamines, polyamides,
peptides, carbohydrates, vitamin moieties, polyethylene glycols,
thioethers, polyethers, cholesterols, thiocholesterols, cholic acid
moieties, folate, lipids, phospholipids, biotin, phenazine,
phenanthridine, anthraquinone, adamantane, acridine, fluoresceins,
rhodamines, coumarins, fluorophores, and dyes.
[0248] In certain embodiments, a conjugate moiety comprises an
active drug substance, for example, aspirin, warfarin,
phenylbutazone, ibuprofen, suprofen, fen-bufen, ketoprofen,
(S)-(+)-pranoprofen, carprofen, dansylsarcosine,
2,3,5-triiodobenzoic acid, fingolimod, flufenamic acid, folinic
acid, a benzothiadiazide, chlorothiazide, a diazepine,
indo-methicin, a barbiturate, a cephalosporin, a sulfa drug, an
antidiabetic, an antibacterial or an antibiotic.
[0249] 2. Conjugate Linkers
[0250] Conjugate moieties are attached to oligonucleotides through
conjugate linkers. In certain oligomeric compounds, the conjugate
linker is a single chemical bond (i.e., the conjugate moiety is
attached directly to an oligonucleotide through a single bond). In
certain embodiments, the conjugate linker comprises a chain
structure, such as a hydrocarbyl chain, or an oligomer of repeating
units such as ethylene glycol, nucleosides, or amino acid
units.
[0251] In certain embodiments, a conjugate linker comprises one or
more groups selected from alkyl, amino, oxo, amide, disulfide,
polyethylene glycol, ether, thioether, and hydroxylamino. In
certain such embodiments, the conjugate linker comprises groups
selected from alkyl, amino, oxo, amide and ether groups. In certain
embodiments, the conjugate linker comprises groups selected from
alkyl and amide groups. In certain embodiments, the conjugate
linker comprises groups selected from alkyl and ether groups. In
certain embodiments, the conjugate linker comprises at least one
phosphorus moiety. In certain embodiments, the conjugate linker
comprises at least one phosphate group. In certain embodiments, the
conjugate linker includes at least one neutral linking group.
[0252] In certain embodiments, conjugate linkers, including the
conjugate linkers described above, are bifunctional linking
moieties, e.g., those known in the art to be useful for attaching
conjugate groups to parent compounds, such as the oligonucleotides
provided herein. In general, a bifunctional linking moiety
comprises at least two functional groups. One of the functional
groups is selected to react with particular site on a parent
compound and the other is selected to react with a conjugate group.
Examples of functional groups used in a bifunctional linking moiety
include but are not limited to electrophiles for reacting with
nucleophilic groups and nucleophiles for reacting with
electrophilic groups. In certain embodiments, bifunctional linking
moieties comprise one or more groups selected from amino, hydroxyl,
carboxylic acid, thiol, alkyl, alkenyl, and alkynyl.
[0253] Examples of conjugate linkers include but are not limited to
pyrrolidine, 8-amino-3,6-dioxaoctanoic acid (ADO), succinimidyl
4-(N-maleimidomethyl) cyclohexane-1-carboxylate (SMCC) and
6-aminohexanoic acid (AHEX or AHA). Other conjugate linkers include
but are not limited to substituted or unsubstituted
C.sub.1-C.sub.10 alkyl, substituted or unsubstituted
C.sub.2-C.sub.10 alkenyl or substituted or unsubstituted
C.sub.2-C.sub.10 alkynyl, wherein a nonlimiting list of preferred
substituent groups includes hydroxyl, amino, alkoxy, carboxy,
benzyl, phenyl, nitro, thiol, thioalkoxy, halogen, alkyl, aryl,
alkenyl and alkynyl.
[0254] In certain embodiments, conjugate linkers comprise 1-10
linker-nucleosides. In certain embodiments, conjugate linkers
comprise 2-5 linker-nucleosides. In certain embodiments, conjugate
linkers comprise exactly 3 linker-nucleosides. In certain
embodiments, conjugate linkers comprise the TCA motif. In certain
embodiments, such linker-nucleosides are modified nucleosides. In
certain embodiments such linker-nucleosides comprise a modified
sugar moiety. In certain embodiments, linker-nucleosides are
unmodified. In certain embodiments, linker-nucleosides comprise an
optionally protected heterocyclic base selected from a purine,
substituted purine, pyrimidine or substituted pyrimidine. In
certain embodiments, a cleavable moiety is a nucleoside selected
from uracil, thymine, cytosine, 4-N-benzoylcytosine, 5-methyl
cytosine, 4-N-benzoyl-5-methyl cytosine, adenine,
6-N-benzoyladenine, guanine and 2-N-isobutyrylguanine. It is
typically desirable for linker-nucleosides to be cleaved from the
oligomeric compound after it reaches a target tissue. Accordingly,
linker-nucleosides are typically linked to one another and to the
remainder of the oligomeric compound through cleavable bonds. In
certain embodiments, such cleavable bonds are phosphodiester
bonds.
[0255] Herein, linker-nucleosides are not considered to be part of
the oligonucleotide. Accordingly, in embodiments in which an
oligomeric compound comprises an oligonucleotide consisting of a
specified number or range of linked nucleosides and/or a specified
percent complementarity to a reference nucleic acid and the
oligomeric compound also comprises a conjugate group comprising a
conjugate linker comprising linker-nucleosides, those
linker-nucleosides are not counted toward the length of the
oligonucleotide and are not used in determining the percent
complementarity of the oligonucleotide for the reference nucleic
acid. For example, an oligomeric compound may comprise (1) a
modified oligonucleotide consisting of 8-30 nucleosides and (2) a
conjugate group comprising 1-10 linker-nucleosides that are
contiguous with the nucleosides of the modified oligonucleotide.
The total number of contiguous linked nucleosides in such an
oligomeric compound is more than 30. Alternatively, an oligomeric
compound may comprise a modified oligonucleotide consisting of 8-30
nucleosides and no conjugate group. The total number of contiguous
linked nucleosides in such an oligomeric compound is no more than
30. Unless otherwise indicated conjugate linkers comprise no more
than 10 linker-nucleosides. In certain embodiments, conjugate
linkers comprise no more than 5 linker-nucleosides. In certain
embodiments, conjugate linkers comprise no more than 3
linker-nucleosides. In certain embodiments, conjugate linkers
comprise no more than 2 linker-nucleosides. In certain embodiments,
conjugate linkers comprise no more than 1 linker-nucleoside.
[0256] In certain embodiments, it is desirable for a conjugate
group to be cleaved from the oligonucleotide. For example, in
certain circumstances oligomeric compounds comprising a particular
conjugate moiety are better taken up by a particular cell type, but
once the oligomeric compound has been taken up, it is desirable
that the conjugate group be cleaved to release the unconjugated or
parent oligonucleotide. Thus, certain conjugate linkers may
comprise one or more cleavable moieties. In certain embodiments, a
cleavable moiety is a cleavable bond. In certain embodiments, a
cleavable moiety is a group of atoms comprising at least one
cleavable bond. In certain embodiments, a cleavable moiety
comprises a group of atoms having one, two, three, four, or more
than four cleavable bonds. In certain embodiments, a cleavable
moiety is selectively cleaved inside a cell or subcellular
compartment, such as a lysosome. In certain embodiments, a
cleavable moiety is selectively cleaved by endogenous enzymes, such
as nucleases.
[0257] In certain embodiments, a cleavable bond is selected from
among: an amide, an ester, an ether, one or both esters of a
phosphodiester, a phosphate ester, a carbamate, or a disulfide. In
certain embodiments, a cleavable bond is one or both of the esters
of a phosphodiester. In certain embodiments, a cleavable moiety
comprises a phosphate or phosphodiester. In certain embodiments,
the cleavable moiety is a phosphodiester linkage between an
oligonucleotide and a conjugate moiety or conjugate group.
[0258] In certain embodiments, a cleavable moiety comprises or
consists of one or more linker-nucleosides. In certain such
embodiments, the one or more linker-nucleosides are linked to one
another and/or to the remainder of the oligomeric compound through
cleavable bonds. In certain embodiments, such cleavable bonds are
unmodified phosphodiester bonds. In certain embodiments, a
cleavable moiety is 2'-deoxy nucleoside that is attached to either
the 3' or 5'-terminal nucleoside of an oligonucleotide by a
phosphate internucleoside linkage and covalently attached to the
remainder of the conjugate linker or conjugate moiety by a
phosphate or phosphorothioate linkage. In certain such embodiments,
the cleavable moiety is 2'-deoxyadenosine.
[0259] 3. Certain Cell-Targeting Conjugate Moieties
[0260] In certain embodiments, a conjugate group comprises a
cell-targeting conjugate moiety. In certain embodiments, a
conjugate group has the general formula:
##STR00012##
[0261] wherein n is from 1 to about 3, m is 0 when n is 1, m is 1
when n is 2 or greater, j is 1 or 0, and k is 1 or 0.
[0262] In certain embodiments, n is 1, j is 1 and k is 0. In
certain embodiments, n is 1, j is 0 and k is 1. In certain
embodiments, n is 1, j is 1 and k is 1. In certain embodiments, n
is 2, j is 1 and k is 0. In certain embodiments, n is 2, j is 0 and
k is 1. In certain embodiments, n is 2, j is 1 and k is 1. In
certain embodiments, n is 3, j is 1 and k is 0. In certain
embodiments, n is 3, j is 0 and k is 1. In certain embodiments, n
is 3, j is 1 and k is 1.
[0263] In certain embodiments, conjugate groups comprise
cell-targeting moieties that have at least one tethered ligand. In
certain embodiments, cell-targeting moieties comprise two tethered
ligands covalently attached to a branching group. In certain
embodiments, cell-targeting moieties comprise three tethered
ligands covalently attached to a branching group.
[0264] In certain embodiments, the cell-targeting moiety comprises
a branching group comprising one or more groups selected from
alkyl, amino, oxo, amide, disulfide, polyethylene glycol, ether,
thioether and hydroxylamino groups. In certain embodiments, the
branching group comprises a branched aliphatic group comprising
groups selected from alkyl, amino, oxo, amide, disulfide,
polyethylene glycol, ether, thioether and hydroxylamino groups. In
certain such embodiments, the branched aliphatic group comprises
groups selected from alkyl, amino, oxo, amide and ether groups. In
certain such embodiments, the branched aliphatic group comprises
groups selected from alkyl, amino and ether groups. In certain such
embodiments, the branched aliphatic group comprises groups selected
from alkyl and ether groups. In certain embodiments, the branching
group comprises a mono or polycyclic ring system.
[0265] In certain embodiments, each tether of a cell-targeting
moiety comprises one or more groups selected from alkyl,
substituted alkyl, ether, thioether, disulfide, amino, oxo, amide,
phosphodiester, and polyethylene glycol, in any combination. In
certain embodiments, each tether is a linear aliphatic group
comprising one or more groups selected from alkyl, ether,
thioether, disulfide, amino, oxo, amide, and polyethylene glycol,
in any combination. In certain embodiments, each tether is a linear
aliphatic group comprising one or more groups selected from alkyl,
phosphodiester, ether, amino, oxo, and amide, in any combination.
In certain embodiments, each tether is a linear aliphatic group
comprising one or more groups selected from alkyl, ether, amino,
oxo, and amid, in any combination. In certain embodiments, each
tether is a linear aliphatic group comprising one or more groups
selected from alkyl, amino, and oxo, in any combination. In certain
embodiments, each tether is a linear aliphatic group comprising one
or more groups selected from alkyl and oxo, in any combination. In
certain embodiments, each tether is a linear aliphatic group
comprising one or more groups selected from alkyl and
phosphodiester, in any combination. In certain embodiments, each
tether comprises at least one phosphorus linking group or neutral
linking group. In certain embodiments, each tether comprises a
chain from about 6 to about 20 atoms in length. In certain
embodiments, each tether comprises a chain from about 10 to about
18 atoms in length. In certain embodiments, each tether comprises
about 10 atoms in chain length.
[0266] In certain embodiments, each ligand of a cell-targeting
moiety has an affinity for at least one type of receptor on a
target cell. In certain embodiments, each ligand has an affinity
for at least one type of receptor on the surface of a mammalian
liver cell. In certain embodiments, each ligand has an affinity for
the hepatic asialoglycoprotein receptor (ASGP-R). In certain
embodiments, each ligand is a carbohydrate. In certain embodiments,
each ligand is, independently selected from galactose, N-acetyl
galactosamine (GalNAc), mannose, glucose, glucosamine and fucose.
In certain embodiments, each ligand is N-acetyl galactosamine
(GalNAc). In certain embodiments, the cell-targeting moiety
comprises 3 GalNAc ligands. In certain embodiments, the
cell-targeting moiety comprises 2 GalNAc ligands. In certain
embodiments, the cell-targeting moiety comprises 1 GalNAc
ligand.
[0267] In certain embodiments, each ligand of a cell-targeting
moiety is a carbohydrate, carbohydrate derivative, modified
carbohydrate, polysaccharide, modified polysaccharide, or
polysaccharide derivative. In certain such embodiments, the
conjugate group comprises a carbohydrate cluster (see, e.g., Maier
et al., "Synthesis of Antisense Oligonucleotides Conjugated to a
Multivalent Carbohydrate Cluster for Cellular Targeting,"
Bioconjugate Chemistry, 2003, 14, 18-29, or Rensen et al., "Design
and Synthesis of Novel N-Acetylgalactosamine-Terminated Glycolipids
for Targeting of Lipoproteins to the Hepatic Asiaglycoprotein
Receptor," J. Med. Chem. 2004, 47, 5798-5808, which are
incorporated herein by reference in their entirety). In certain
such embodiments, each ligand is an amino sugar or a thio sugar.
For example, amino sugars may be selected from any number of
compounds known in the art, such as sialic acid,
.alpha.-D-galactosamine, .beta.-muramic acid,
2-deoxy-2-methylamino-L-glucopyranose,
4,6-dideoxy-4-formamido-2,3-di-O-methyl-D-mannopyranose,
2-deoxy-2-sulfoamino-D-glucopyranose and N-sulfo-D-glucosamine, and
N-glycoloyl-.alpha.-neuraminic acid. For example, thio sugars may
be selected from 5-Thio-.beta.-D-glucopyranose, methyl
2,3,4-tri-O-acetyl-1-thio-6-O-trityl-.alpha.-D-glucopyranoside,
4-thio-.beta.-D-galactopyranose, and ethyl
3,4,6,7-tetra-O-acetyl-2-deoxy-1,5-dithio-.alpha.-D-gluco-heptopyranoside-
.
[0268] In certain embodiments, conjugate groups comprise a
cell-targeting moiety having the formula:
##STR00013##
[0269] In certain embodiments, conjugate groups comprise a
cell-targeting moiety having the formula:
##STR00014##
[0270] In certain embodiments, conjugate groups comprise a
cell-targeting moiety having the formula:
##STR00015##
[0271] In certain embodiments, compounds described herein comprise
a conjugate group described herein as "THA-GalNAac.sub.3".
THA-GalNAc.sub.3 is shown below without the optional cleavable
moiety at the end of the linker region:
##STR00016##
[0272] In certain embodiments, compounds described herein comprise
THA-GalNAc.sub.3-phosphate, also represented as
(THA-GalNAc.sub.3)o, having the formula:
##STR00017## [0273] wherein modified oligonucleotide represents a
modified oligonucleotide.
[0274] Representative publications that teach the preparation of
certain of the above noted conjugate groups and compounds
comprising conjugate groups, tethers, conjugate linkers, branching
groups, ligands, cleavable moieties as well as other modifications
include without limitation, U.S. Pat. Nos. 5,994,517, 6,300,319,
6,660,720, 6,906,182, 7,262,177, 7,491,805, 8,106,022, 7,723,509,
9,127,276, US 2006/0148740, US 2011/0123520, WO 2013/033230 and WO
2012/037254, Biessen et al., J. Med. Chem. 1995, 38, 1846-1852, Lee
et al., Bioorganic & Medicinal Chemistry 2011, 19, 2494-2500,
Rensen et al., J. Biol. Chem. 2001, 276, 37577-37584, Rensen et
al., J. Med. Chem. 2004, 47, 5798-5808, Sliedregt et al., J. Med.
Chem. 1999, 42, 609-618, and Valentijn et al., Tetrahedron, 1997,
53, 759-770, each of which is incorporated by reference herein in
its entirety.
[0275] In certain embodiments, compounds described herein comprise
modified oligonucleotides comprising a gapmer or fully modified
motif and a conjugate group comprising at least one, two, or three
GalNAc ligands. In certain embodiments compounds described herein
comprise a conjugate group found in any of the following
references: Lee, Carbohydr Res, 1978, 67, 509-514; Connolly et al.,
J Biol Chem, 1982, 257, 939-945; Pavia et al., Int J Pep Protein
Res, 1983, 22, 539-548; Lee et al., Biochem, 1984, 23, 4255-4261;
Lee et al., Glycoconjugate J, 1987, 4, 317-328; Toyokuni et al.,
Tetrahedron Lett, 1990, 31, 2673-2676; Biessen et al., J Med Chem,
1995, 38, 1538-1546; Valentijn et al., Tetrahedron, 1997, 53,
759-770; Kim et al., Tetrahedron Lett, 1997, 38, 3487-3490; Lee et
al., Bioconjug Chem, 1997, 8, 762-765; Kato et al., Glycobiol,
2001, 11, 821-829; Rensen et al., J Biol Chem, 2001, 276,
37577-37584; Lee et al., Methods Enzymol, 2003, 362, 38-43;
Westerlind et al., Glycoconj J, 2004, 21, 227-241; Lee et al.,
Bioorg Med Chem Lett, 2006, 16(19), 5132-5135; Maierhofer et al.,
Bioorg Med Chem, 2007, 15, 7661-7676; Khorev et al., Bioorg Med
Chem, 2008, 16, 5216-5231; Lee et al., Bioorg Med Chem, 2011, 19,
2494-2500; Kornilova et al., Analyt Biochem, 2012, 425, 43-46;
Pujol et al., Angew Chemie Int Ed Engl, 2012, 51, 7445-7448;
Biessen et al., J Med Chem, 1995, 38, 1846-1852; Sliedregt et al.,
J Med Chem, 1999, 42, 609-618; Rensen et al., J Med Chem, 2004, 47,
5798-5808; Rensen et al., Arterioscler Thromb Vasc Biol, 2006, 26,
169-175; van Rossenberg et al., Gene Ther, 2004, 11, 457-464; Sato
et al., J Am Chem Soc, 2004, 126, 14013-14022; Lee et al., J Org
Chem, 2012, 77, 7564-7571; Biessen et al., FASEB J, 2000, 14,
1784-1792; Rajur et al., Bioconjug Chem, 1997, 8, 935-940; Duff et
al., Methods Enzymol, 2000, 313, 297-321; Maier et al., Bioconjug
Chem, 2003, 14, 18-29; Jayaprakash et al., Org Lett, 2010, 12,
5410-5413; Manoharan, Antisense Nucleic Acid Drug Dev, 2002, 12,
103-128; Merwin et al., Bioconjug Chem, 1994, 5, 612-620; Tomiya et
al., Bioorg Med Chem, 2013, 21, 5275-5281; International
applications WO1998/013381; WO2011/038356; WO1997/046098;
WO2008/098788; WO2004/101619; WO2012/037254; WO2011/120053;
WO2011/100131; WO2011/163121; WO2012/177947; WO2013/033230;
WO2013/075035; WO2012/083185; WO2012/083046; WO2009/082607;
WO2009/134487; WO2010/144740; WO2010/148013; WO1997/020563;
WO2010/088537; WO2002/043771; WO2010/129709; WO2012/068187;
WO2009/126933; WO2004/024757; WO2010/054406; WO2012/089352;
WO2012/089602; WO2013/166121; WO2013/165816; U.S. Pat. Nos.
4,751,219; 8,552,163; 6,908,903; 7,262,177; 5,994,517; 6,300,319;
8,106,022; 7,491,805; 7,491,805; 7,582,744; 8,137,695; 6,383,812;
6,525,031; 6,660,720; 7,723,509; 8,541,548; 8,344,125; 8,313,772;
8,349,308; 8,450,467; 8,501,930; 8,158,601; 7,262,177; 6,906,182;
6,620,916; 8,435,491; 8,404,862; 7,851,615; Published U.S. Patent
Application Publications US2011/0097264; US2011/0097265;
US2013/0004427; US2005/0164235; US2006/0148740; US2008/0281044;
US2010/0240730; U52003/0119724; US2006/0183886; US2008/0206869;
U52011/0269814; US2009/0286973; US2011/0207799; US2012/0136042;
US2012/0165393; US2008/0281041; U52009/0203135; US2012/0035115;
US2012/0095075; US2012/0101148; US2012/0128760; US2012/0157509;
US2012/0230938; US2013/0109817; US2013/0121954; US2013/0178512;
US2013/0236968; US2011/0123520; US2003/0077829; US2008/0108801; and
US2009/0203132; each of which is incorporated by reference in its
entirety.
[0276] B. Certain Terminal Groups
[0277] In certain embodiments, oligomeric compounds comprise one or
more terminal groups. In certain such embodiments, oligomeric
compounds comprise a stabilized 5'-phosphate. Stabilized
5'-phosphates include, but are not limited to 5'-phosphanates,
including, but not limited to 5'-vinylphosphonates. In certain
embodiments, terminal groups comprise one or more abasic
nucleosides and/or inverted nucleosides. In certain embodiments,
terminal groups comprise one or more 2'-linked nucleosides. In
certain such embodiments, the 2'-linked nucleoside is an abasic
nucleoside.
[0278] III. Oligomeric Duplexes
[0279] In certain embodiments, oligomeric compounds described
herein comprise an oligonucleotide, having a nucleobase sequence
complementary to that of a target nucleic acid. In certain
embodiments, an oligomeric compound is paired with a second
oligomeric compound to form an oligomeric duplex. Such oligomeric
duplexes comprise a first oligomeric compound having a region
complementary to a target nucleic acid and a second oligomeric
compound having a region complementary to the first oligomeric
compound. In certain embodiments, the first oligomeric compound of
an oligomeric duplex comprises or consists of (1) a modified or
unmodified oligonucleotide and optionally a conjugate group and (2)
a second modified or unmodified oligonucleotide and optionally a
conjugate group. Either or both oligomeric compounds of an
oligomeric duplex may comprise a conjugate group. The
oligonucleotides of each oligomeric compound of an oligomeric
duplex may include non-complementary overhanging nucleosides.
[0280] IV. Antisense Activity
[0281] In certain embodiments, oligomeric compounds and oligomeric
duplexes are capable of hybridizing to a target nucleic acid,
resulting in at least one antisense activity; such oligomeric
compounds and oligomeric duplexes are antisense compounds. In
certain embodiments, antisense compounds have antisense activity
when they reduce or inhibit the amount or activity of a target
nucleic acid by 25% or more in the standard cell assay. In certain
embodiments, antisense compounds selectively affect one or more
target nucleic acid. Such antisense compounds comprise a nucleobase
sequence that hybridizes to one or more target nucleic acid,
resulting in one or more desired antisense activity and does not
hybridize to one or more non-target nucleic acid or does not
hybridize to one or more non-target nucleic acid in such a way that
results in significant undesired antisense activity.
[0282] In certain antisense activities, hybridization of an
antisense compound to a target nucleic acid results in recruitment
of a protein that cleaves the target nucleic acid. For example,
certain antisense compounds result in RNase H mediated cleavage of
the target nucleic acid. RNase H is a cellular endonuclease that
cleaves the RNA strand of an RNA:DNA duplex. The DNA in such an
RNA:DNA duplex need not be unmodified DNA. In certain embodiments,
antisense compounds described herein are sufficiently "DNA-like"to
elicit RNase H activity. In certain embodiments, one or more
non-DNA-like nucleoside in the gap of a gapmer is tolerated.
[0283] In certain antisense activities, an antisense compound or a
portion of an antisense compound is loaded into an RNA-induced
silencing complex (RISC), ultimately resulting in cleavage of the
target nucleic acid. For example, certain antisense compounds
result in cleavage of the target nucleic acid by Argonaute.
Antisense compounds that are loaded into RISC are RNAi compounds.
RNAi compounds may be double-stranded (siRNA) or single-stranded
(ssRNA).
[0284] In certain embodiments, hybridization of an antisense
compound to a target nucleic acid does not result in recruitment of
a protein that cleaves that target nucleic acid. In certain
embodiments, hybridization of the antisense compound to the target
nucleic acid results in alteration of splicing of the target
nucleic acid.
[0285] In certain embodiments, hybridization of an antisense
compound to a target nucleic acid results in inhibition of a
binding interaction between the target nucleic acid and a protein
or other nucleic acid. In certain embodiments, hybridization of an
antisense compound to a target nucleic acid results in alteration
of translation of the target nucleic acid.
[0286] Antisense activities may be observed directly or indirectly.
In certain embodiments, observation or detection of an antisense
activity involves observation or detection of a change in an amount
of a target nucleic acid or protein encoded by such target nucleic
acid, a change in the ratio of splice variants of a nucleic acid or
protein and/or a phenotypic change in a cell or animal.
[0287] V. Certain Target Nucleic Acids
[0288] In certain embodiments, oligomeric compounds comprise or
consist of an oligonucleotide comprising a region that is
complementary to a target nucleic acid. In certain embodiments, the
target nucleic acid is an endogenous RNA molecule. In certain
embodiments, the target nucleic acid encodes a protein. In certain
such embodiments, the target nucleic acid is selected from: a
mature mRNA and a pre-mRNA, including intronic, exonic and
untranslated regions. In certain embodiments, the target RNA is a
mature mRNA. In certain embodiments, the target nucleic acid is a
pre-mRNA. In certain such embodiments, the target region is
entirely within an intron. In certain embodiments, the target
region spans an intron/exon junction. In certain embodiments, the
target region is at least 50% within an intron. In certain
embodiments, the target nucleic acid is the RNA transcriptional
product of a retrogene. In certain embodiments, the target nucleic
acid is a non-coding RNA. In certain such embodiments, the target
non-coding RNA is selected from: a long non-coding RNA, a short
non-coding RNA, an intronic RNA molecule.
[0289] A. Complementarity/Mismatches to the Target Nucleic Acid
[0290] It is possible to introduce mismatch bases without
eliminating activity. For example, Gautschi et al (J. Natl. Cancer
Inst. 93:463-471, March 2001) demonstrated the ability of an
oligonucleotide having 100% complementarity to the bcl-2 mRNA and
having 3 mismatches to the bcl-xL mRNA to reduce the expression of
both bcl-2 and bcl-xL in vitro and in vivo. Furthermore, this
oligonucleotide demonstrated potent anti-tumor activity in vivo.
Maher and Dolnick (Nuc. Acid. Res. 16:3341-3358, 1988) tested a
series of tandem 14 nucleobase oligonucleotides, and a 28 and 42
nucleobase oligonucleotides comprised of the sequence of two or
three of the tandem oligonucleotides, respectively, for their
ability to arrest translation of human DHFR in a rabbit
reticulocyte assay. Each of the three 14 nucleobase
oligonucleotides alone was able to inhibit translation, albeit at a
more modest level than the 28 or 42 nucleobase
oligonucleotides.
[0291] In certain embodiments, oligonucleotides are complementary
to the target nucleic acid over the entire length of the
oligonucleotide. In certain embodiments, oligonucleotides are 99%,
95%, 90%, 85%, or 80% complementary to the target nucleic acid. In
certain embodiments, oligonucleotides are at least 80%
complementary to the target nucleic acid over the entire length of
the oligonucleotide and comprise a region that is 100% or fully
complementary to a target nucleic acid. In certain embodiments, the
region of full complementarity is from 6 to 20, 10 to 18, or 18 to
20 nucleobases in length.
[0292] In certain embodiments, oligonucleotides comprise one or
more mismatched nucleobases relative to the target nucleic acid. In
certain embodiments, antisense activity against the target is
reduced by such mismatch, but activity against a non-target is
reduced by a greater amount. Thus, in certain embodiments
selectivity of the oligonucleotide is improved. In certain
embodiments, the mismatch is specifically positioned within an
oligonucleotide having a gapmer motif. In certain embodiments, the
mismatch is at position 1, 2, 3, 4, 5, 6, 7, or 8 from the 5'-end
of the gap region. In certain embodiments, the mismatch is at
position 9, 8, 7, 6, 5, 4, 3, 2, 1 from the 3'-end of the gap
region. In certain embodiments, the mismatch is at position 1, 2,
3, or 4 from the 5'-end of the wing region. In certain embodiments,
the mismatch is at position 4, 3, 2, or 1 from the 3'-end of the
wing region.
[0293] B. FXI
[0294] In certain embodiments, oligomeric compounds comprise or
consist of an oligonucleotide comprising a region that is
complementary to a target nucleic acid, wherein the target nucleic
acid is FXI. In certain embodiments, FXI nucleic acid has the
sequence set forth in SEQ ID NO: 1 (the complement of GENBANK
Accession No: NT 022792.17 truncated from nucleobase 19598000 to
Ser. No. 19/624,000) and SEQ ID NO: 2 (GENBANK Accession No:
NM_000128.3).
[0295] In certain embodiments, methods comprise contacting a cell
with an oligomeric compound disclosed herein. In certain
embodiments, methods comprise administering an oligomeric compound
disclosed herein to an animal, thereby contacting a cell in the
animal. In certain embodiments, contacting a cell with an
oligomeric compound comprising a modified oligonucleotide
complementary to SEQ ID NO: 1 or SEQ ID NO: 2 reduces the amount of
FXI RNA, and in certain embodiments reduces the amount of FXI
protein. In certain embodiments, the oligomeric compound consists
of a conjugate group attached to the 5' end of a modified
oligonucleotide. In certain embodiments, contacting a cell in an
animal with an oligomeric compound comprising a modified
oligonucleotide complementary to SEQ ID NO: 1 or SEQ ID NO: 2
treats, prevents, or ameliorates a thromboembolic condition. In
certain embodiments, the thromboembolic condition is deep vein
thrombosis, venous or arterial thrombosis, pulmonary embolism,
myocardial infarction, stroke, thrombosis associated with chronic
kidney disease or end-stage renal disease (ESRD), including
thrombosis associated with dialysis, or other procoagulant
condition. In certain embodiments, the oligomeric compound consists
of a conjugate group attached to the 5' end of a modified
oligonucleotide. In certain embodiments, contacting a cell in an
animal with an oligomeric compound comprising a modified
oligonucleotide complementary to SEQ ID NO: 1 or SEQ ID NO: 2
treats, prevents, or ameliorates a thromboembolic condition without
increasing bleeding risk. In certain embodiments, the
thromboembolic condition is deep vein thrombosis, venous or
arterial thrombosis, pulmonary embolism, myocardial infarction,
stroke, thrombosis associated with chronic kidney disease or
end-stage renal disease (ESRD), including thrombosis associated
with dialysis, or other procoagulant condition. In certain
embodiments, the oligomeric compound consists of a conjugate group
attached to the 5' end of a modified oligonucleotide.
[0296] A FXI RNA may be quantified, e.g., by quantitative PCR. FXI
proteins may be quantified with standard protein quantification
tests, e.g., ELISA. The FXI protein may be an inactive form
(zymogen). The FXI protein may be an active form (FXIa). The active
form of FXI may promote converting FIX from its inactive form (FIX)
to its active form (FIXa). FXI activity may be assessed with a
blood test that characterizes coagulation of blood. Non-limiting
examples of such blood tests are a partial thromboplastin time
(PTT) test or activated partial thromboplastin time test (aPTT or
APTT). FXI activity in a test plasma sample may be assayed by
adding the test plasma sample to a plasma sample that is
immunodepleted of FXI and comparing clotting of the resulting
combined sample to a reference sample with a reference amount of
FXI.
[0297] In certain embodiments, contacting a cell in an animal with
an oligomeric compound comprising a modified oligonucleotide
complementary to SEQ ID NO: 1 or SEQ ID NO: 2 treats, prevents, or
ameliorates a thromboembolic condition. In certain embodiments, the
thromboembolic condition is deep vein thrombosis, venous or
arterial thrombosis, pulmonary embolism, myocardial infarction,
stroke, thrombosis associated with chronic kidney disease or
end-stage renal disease (ESRD), including thrombosis associated
with dialysis, or other procoagulant condition. In certain
embodiments, the oligomeric compound consists of a conjugate group
attached to the 5' end of a modified oligonucleotide. In certain
embodiments, contacting a cell in an animal with an oligomeric
compound comprising a modified oligonucleotide complementary to SEQ
ID NO: 1 or SEQ ID NO: 2 treats, prevents, or ameliorates a
thromboembolic condition without increasing bleeding risk. In
certain embodiments, the thromboembolic condition is deep vein
thrombosis, venous or arterial thrombosis, pulmonary embolism,
myocardial infarction, stroke, thrombosis associated with chronic
kidney disease or end-stage renal disease (ESRD), including
thrombosis associated with dialysis, or other procoagulant
condition. In certain embodiments, the oligomeric compound consists
or consists essentially of a conjugate group attached to the 5' end
of a modified oligonucleotide.
[0298] C. Certain Target Nucleic Acids in Certain Tissues
[0299] In certain embodiments, oligomeric compounds comprise or
consist of an oligonucleotide comprising a region that is
complementary to a target nucleic acid, wherein the target nucleic
acid is expressed in a pharmacologically relevant tissue. In
certain embodiments, the pharmacologically relevant tissues are the
cells and tissues that comprise the alimentary and/or excretory
system. Such cells and tissues include the liver, kidney, and
pancreas.
[0300] VI. Certain Pharmaceutical Compositions
[0301] In certain embodiments, pharmaceutical compositions
described herein comprise one or more oligomeric compounds. In
certain embodiments, the one or more oligomeric compounds each
comprise a modified oligonucleotide. In certain embodiments, the
pharmaceutical composition comprises a pharmaceutically acceptable
diluent or carrier. In certain embodiments, a pharmaceutical
composition comprises a sterile saline solution and one or more
oligomeric compounds. In certain embodiments, a pharmaceutical
composition consists or consists essentially of a sterile saline
solution and one or more oligomeric compounds. In certain
embodiments, the sterile saline is pharmaceutical grade saline. In
certain embodiments, a pharmaceutical composition comprises one or
more oligomeric compounds and sterile water. In certain
embodiments, a pharmaceutical composition consists or consists
essentially of one or more oligomeric compounds and sterile water.
In certain embodiments, the sterile water is pharmaceutical grade
water. In certain embodiments, the pharmaceutically acceptable
diluent or carrier is distilled water for injection. In certain
embodiments, a pharmaceutical composition comprises one or more
oligomeric compound and phosphate-buffered saline (PBS). In certain
embodiments, a pharmaceutical composition consists or consists
essentially of one or more oligomeric compounds and PBS. In certain
embodiments, the sterile PBS is pharmaceutical grade PBS. In
certain embodiments, a pharmaceutical composition comprises one or
more oligomeric compound and artificial cerebrospinal fluid. In
certain embodiments, the sterile PBS is pharmaceutical grade PBS.
In certain embodiments, a pharmaceutical composition consists or
consists essentially of artificial cerebrospinal fluid. In certain
embodiments, the artificial cerebrospinal fluid is pharmaceutical
grade.
[0302] In certain embodiments, pharmaceutical compositions comprise
one or more oligomeric compounds disclosed herein and one or more
excipients. In certain embodiments, excipients are selected from
water, salt solutions, alcohol, polyethylene glycols, gelatin,
lactose, amylase, magnesium stearate, talc, silicic acid, viscous
paraffin, hydroxymethylcellulose and polyvinylpyrrolidone.
[0303] In certain embodiments, oligomeric compounds may be admixed
with pharmaceutically acceptable active and/or inert substances for
the preparation of pharmaceutical compositions or formulations.
Compositions and methods for the formulation of pharmaceutical
compositions depend on a number of criteria, including, but not
limited to, route of administration, extent of disease, or dose to
be administered.
[0304] In certain embodiments, pharmaceutical compositions
comprising an oligomeric compound disclosed herein encompass any
pharmaceutically acceptable salts of the oligomeric compound,
esters of the oligomeric compound, or salts of such esters. In
certain embodiments, pharmaceutical compositions comprising
oligomeric compounds comprising one or more oligonucleotide, upon
administration to an animal, including a human, are capable of
providing (directly or indirectly) the biologically active
metabolite or residue thereof. Accordingly, for example, the
disclosure is also drawn to pharmaceutically acceptable salts of
oligomeric compounds, prodrugs, pharmaceutically acceptable salts
of such prodrugs, and other bioequivalents. Suitable
pharmaceutically acceptable salts include, but are not limited to,
sodium and potassium salts. In certain embodiments, prodrugs
comprise one or more conjugate group attached to an
oligonucleotide, wherein the conjugate group is cleaved by
endogenous nucleases within the body.
[0305] Lipid moieties have been used in nucleic acid therapies in a
variety of methods. In certain such methods, the nucleic acid, such
as an oligomeric compound, is introduced into preformed liposomes
or lipoplexes made of mixtures of cationic lipids and neutral
lipids. In certain methods, DNA complexes with mono- or
poly-cationic lipids are formed without the presence of a neutral
lipid. In certain embodiments, a lipid moiety is selected to
increase distribution of a pharmaceutical agent to a particular
cell or tissue. In certain embodiments, a lipid moiety is selected
to increase distribution of a pharmaceutical agent to fat tissue.
In certain embodiments, a lipid moiety is selected to increase
distribution of a pharmaceutical agent to muscle tissue.
[0306] In certain embodiments, pharmaceutical compositions
disclosed herein comprise a delivery system. Examples of delivery
systems include, but are not limited to, liposomes and emulsions.
Certain delivery systems are useful for preparing certain
pharmaceutical compositions including those comprising hydrophobic
compounds. In certain embodiments, certain organic solvents such as
dimethylsulfoxide are used.
[0307] In certain embodiments, pharmaceutical compositions comprise
one or more tissue-specific delivery molecules designed to deliver
oligomeric compounds described herein to specific tissues or cell
types. For example, in certain embodiments, pharmaceutical
compositions include liposomes coated with a tissue-specific
antibody.
[0308] In certain embodiments, pharmaceutical compositions comprise
a co-solvent system. Certain of such co-solvent systems comprise,
for example, benzyl alcohol, a nonpolar surfactant, a
water-miscible organic polymer, and an aqueous phase. In certain
embodiments, such co-solvent systems are used for hydrophobic
compounds. A non-limiting example of such a co-solvent system is
the VPD co-solvent system, which is a solution of absolute ethanol
comprising 3% w/v benzyl alcohol, 8% w/v of the nonpolar surfactant
Polysorbate 80.TM. and 65% w/v polyethylene glycol 300. The
proportions of such co-solvent systems may be varied considerably
without significantly altering their solubility and toxicity
characteristics. Furthermore, the identity of co-solvent components
may be varied: for example, other surfactants may be used instead
of Polysorbate 80.TM.; the fraction size of polyethylene glycol may
be varied; other biocompatible polymers may replace polyethylene
glycol, e.g., polyvinyl pyrrolidone; and other sugars or
polysaccharides may substitute for dextrose.
[0309] In certain embodiments, pharmaceutical compositions
disclosed herein are prepared for oral administration. In certain
embodiments, pharmaceutical compositions are prepared for buccal
administration. In certain embodiments, a pharmaceutical
composition is prepared for administration by injection (e.g.,
intravenous, subcutaneous, intramuscular, intrathecal (IT),
intracerebroventricular (ICV), etc.). In certain of such
embodiments, a pharmaceutical composition comprises a carrier and
is formulated in aqueous solution, such as water or physiologically
compatible buffers such as Hanks's solution, Ringer's solution, or
physiological saline buffer. In certain embodiments, other
ingredients are included (e.g., ingredients that aid in solubility
or serve as preservatives). In certain embodiments, injectable
suspensions are prepared using appropriate liquid carriers,
suspending agents and the like. Certain pharmaceutical compositions
for injection are presented in unit dosage form, e.g., in ampoules
or in multi-dose containers. Certain pharmaceutical compositions
for injection are suspensions, solutions or emulsions in oily or
aqueous vehicles, and may contain formulatory agents such as
suspending, stabilizing and/or dispersing agents. Certain solvents
suitable for use in pharmaceutical compositions for injection
include, but are not limited to, lipophilic solvents and fatty
oils, such as sesame oil, synthetic fatty acid esters, such as
ethyl oleate or triglycerides, and liposomes. Aqueous injection
suspensions may contain.
[0310] Under certain conditions, certain compounds disclosed herein
act as acids. Although such compounds may be drawn or described in
protonated (free acid) form, in ionized (anion) form, or ionized
and in association with a cation (salt) form, aqueous solutions of
such compounds exist in equilibrium among such forms. For example,
a phosphate linkage of an oligonucleotide in aqueous solution
exists in equilibrium among free acid, anion, and salt forms.
Unless otherwise indicated, compounds described herein are intended
to include all such forms. Moreover, certain oligonucleotides have
several such linkages, each of which is in equilibrium. Thus,
oligonucleotides in solution exist in an ensemble of forms at
multiple positions all at equilibrium. The term "oligonucleotide"
is intended to include all such forms. Drawn structures necessarily
depict a single form. Nevertheless, unless otherwise indicated,
such drawings are likewise intended to include corresponding forms.
Herein, a structure depicting the free acid of a compound followed
by the term "or salts thereof" expressly includes all such forms
that may be fully or partially protonated/de-protonated/in
association with a cation. In certain instances, one or more
specific cation is identified.
[0311] In certain embodiments, oligomeric compounds disclosed
herein are in aqueous solution with sodium. In certain embodiments,
oligomeric compounds are in aqueous solution with potassium. In
certain embodiments, oligomeric compounds are in PBS. In certain
embodiments, oligomeric compounds are in water. In certain such
embodiments, the pH of the solution is adjusted with NaOH and/or
HCl to achieve a desired pH.
[0312] Herein, certain specific doses are described. A dose may be
in the form of a dosage unit. For clarity, a dose (or dosage unit)
of an oligomeric compound in milligrams indicates the mass of the
free acid form of the oligomeric compound. As described above, in
aqueous solution, the free acid is in equilibrium with anionic and
salt forms. However, for the purpose of calculating dose, it is
assumed that the oligomeric compound exists as a solvent-free,
sodium-acetate free, anhydrous, conjugated free acid. For example,
where an oligomeric compound is in solution comprising sodium
(e.g., saline), the oligomeric compound may be partially or fully
de-protonated and in association with Na.sup.+ ions. However, the
mass of the protons are nevertheless counted toward the weight of
the dose, and the mass of the Na.sup.+ ions are not counted toward
the weight of the dose. Thus, for example, a dose, or dosage unit,
of 80 mg of Compound No. 957943 equals the number of fully
protonated molecules that weighs 80 mg. This would be equivalent to
84 mg of solvent-free, sodium-acetate free, anhydrous sodiated
Compound No. 957943. When an oligomeric compound comprises a
conjugate, the mass of conjugate is included in calculating the
dose of such oligomeric compound.
[0313] VII. Certain Dosage Amounts
[0314] In certain embodiments, pharmaceutical compositions
described herein are administered in the form of a dosage unit. The
dosage unit may be prepared for injection. The dosage unit may be
prepared for infusion. In certain embodiments, the dosage unit
comprises an oligomeric compound disclosed herein and a
pharmaceutically acceptable carrier or diluent. In certain
embodiments, the dosage unit consists or consists essentially of an
oligomeric compound disclosed herein and a pharmaceutically
acceptable carrier or diluent. In certain embodiments, the
oligomeric compound comprises a modified oligonucleotide having the
nucleobase sequence of SEQ ID NO: 3. In certain embodiments, the
oligomeric compound is Compound No. 957943.
[0315] In certain embodiments, the oligomeric compound is present
in the dosage unit at an amount selected from 5 mg, 10 mg, 15 mg,
20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg, 65
mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 95 mg, 100 mg, 105 mg, 110
mg, 115 mg, 120 mg, 125 mg, 130 mg, 135 mg, 140 mg, 145 mg, 150 mg,
155 mg, 160 mg, 165 mg, 170 mg, 175 mg, 180 mg, 185 mg, 190 mg, 195
mg, 200 mg, 205 mg, 210 mg, 215 mg, 220 mg, 225 mg, 230 mg, 235 mg,
240 mg, 245 mg, 250 mg, 255 mg, 260 mg, 265 mg, 270 mg, 275 mg, 280
mg, 285 mg, 290 mg, 295 mg, and 300 mg. In certain embodiments, the
oligomeric compound is present in the dosage unit at an amount
selected from about 5 mg, about 10 mg, about 15 mg, about 20 mg,
about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg,
about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg,
about 75 mg, about 80 mg, about 85 mg, about 90 mg, about 95 mg,
about 100 mg, about 105 mg, about 110 mg, about 115 mg, about 120
mg, about 125 mg, about 130 mg, about 135 mg, about 140 mg, about
145 mg, about 150 mg, about 155 mg, about 160 mg, about 165 mg,
about 170 mg, about 175 mg, about 180 mg, about 185 mg, about 190
mg, about 195 mg, about 200 mg, about 205 mg, about 210 mg, about
215 mg, about 220 mg, about 225 mg, about 230 mg, about 235 mg,
about 240 mg, about 245 mg, about 250 mg, about 255 mg, about 260
mg, about 265 mg, about 270 mg, about 275 mg, about 280 mg, about
285 mg, about 290 mg, about 295 mg, and about 300 mg.
[0316] In certain embodiments, the oligomeric compound is present
in the dosage unit at an amount selected from 20 mg, 21 mg, 22 mg,
23 mg, 24 mg, 25 mg, 26 mg, 27 mg, 28 mg, 29 mg, 30 mg, 31 mg, 32
mg, 33 mg, 34 mg, 35 mg, 36 mg, 37 mg, 38 mg, 39 mg, 40 mg, 41 mg,
42 mg, 43 mg, 44 mg, 45 mg, 46 mg, 47 mg, 48 mg, 49 mg, 50 mg, 51
mg, 52 mg, 53 mg, 54 mg, 55 mg, 56 mg, 57 mg, 58 mg, 59 mg, 60 mg,
61 mg, 62 mg, 63 mg, 64 mg, 65 mg, 66 mg, 67 mg, 68 mg, 69 mg, 70
mg, 71 mg, 72 mg, 73 mg, 74 mg, 75 mg, 76 mg, 77 mg, 78 mg, 79 mg,
80 mg, 81 mg, 82 mg, 83 mg, 84 mg, 85 mg, 86 mg, 87 mg, 88 mg, 89
mg, 90 mg, 91 mg, 92 mg, 93 mg, 94 mg, 95 mg, 96 mg, 97 mg, 98 mg,
99 mg, 100 mg, 101 mg, 102 mg, 103 mg, 104 mg, 105 mg, 106 mg, 107
mg, 108 mg, 109 mg, 110 mg, 111 mg, 112 mg, 113 mg, 114 mg, 115 mg,
116 mg, 117 mg, 118 mg, 119 mg, 120 mg, 121 mg, 122 mg, 123 mg, 124
mg, 125 mg, 126 mg, 127 mg, 128 mg, 129 mg, 130 mg, 131 mg, 132 mg,
133 mg, 134 mg, 135 mg, 136 mg, 137 mg, 138 mg, 139 mg, and 140 mg.
In certain embodiments, the oligomeric compound is present in the
dosage unit at an amount selected from about 20 mg, about 21 mg,
about 22 mg, about 23 mg, about 24 mg, about 25 mg, about 26 mg,
about 27 mg, about 28 mg, about 29 mg, about 30 mg, about 31 mg,
about 32 mg, about 33 mg, about 34 mg, about 35 mg, about 36 mg,
about 37 mg, about 38 mg, about 39 mg, about 40 mg, about 41 mg,
about 42 mg, about 43 mg, about 44 mg, about 45 mg, about 46 mg,
about 47 mg, about 48 mg, about 49 mg, about 50 mg, about 51 mg,
about 52 mg, about 53 mg, about 54 mg, about 55 mg, about 56 mg,
about 57 mg, about 58 mg, about 59 mg, about 60 mg, about 61 mg,
about 62 mg, about 63 mg, about 64 mg, about 65 mg, about 66 mg,
about 67 mg, about 68 mg, about 69 mg, about 70 mg, about 71 mg,
about 72 mg, about 73 mg, about 74 mg, about 75 mg, about 76 mg,
about 77 mg, about 78 mg, about 79 mg, about 80 mg, about 81 mg,
about 82 mg, about 83 mg, about 84 mg, about 85 mg, about 86 mg,
about 87 mg, about 88 mg, about 89 mg, about 90 mg, about 91 mg,
about 92 mg, about 93 mg, about 94 mg, about 95 mg, about 96 mg,
about 97 mg, about 98 mg, about 99 mg, about 100 mg, about 101 mg,
about 102 mg, about 103 mg, about 104 mg, about 105 mg, about 106
mg, about 107 mg, about 108 mg, about 109 mg, about 110 mg, about
111 mg, about 112 mg, about 113 mg, about 114 mg, about 115 mg,
about 116 mg, about 117 mg, about 118 mg, about 119 mg, about 120
mg, about 121 mg, about 122 mg, about 123 mg, about 124 mg, about
125 mg, about 126 mg, about 127 mg, about 128 mg, about 129 mg,
about 130 mg, about 131 mg, about 132 mg, about 133 mg, about 134
mg, about 135 mg, about 136 mg, about 137 mg, about 138 mg, about
139 mg, and about 140 mg.
[0317] In certain embodiments, the oligomeric compound is present
in the dosage unit at an amount selected from 75.0 mg, 75.1 mg,
75.2 mg, 75.3 mg, 75.4 mg, 75.5 mg, 75.6 mg, 75.7 mg, 75.8 mg, 75.9
mg, 76.0 mg, 76.1 mg, 76.2 mg, 76.3 mg. 76.4 mg, 76.5 mg, 76.6 mg,
76.7 mg, 76.8 mg, 76.9 mg, 77.0 mg, 77.1 mg, 77.2 mg, 77.3 mg, 77.4
mg, 77.5 mg, 77.6 mg, 77.7 mg, 77.8 mg, 77.9 mg, 78.0 mg, 78.1 mg,
78.2 mg, 78.3 mg. 78.4 mg, 78.5 mg, 78.6 mg, 78.7 mg, 78.8 mg, 78.9
mg, 79.0 mg, 79.1 mg, 79.2 mg, 79.3 mg, 79.4 mg, 79.5 mg, 79.6 mg,
79.7 mg, 79.8 mg, 79.9 mg, 80.0 mg, 80.1 mg, 80.2 mg, 80.3 mg. 80.4
mg, 80.5 mg, 80.6 mg, 80.7 mg, 80.8 mg, 80.9 mg, 81.0 mg, 81.1 mg,
81.2 mg, 81.3 mg, 81.4 mg, 81.5 mg, 81.6 mg, 81.7 mg, 81.8 mg, 81.9
mg, 82.0 mg, 82.1 mg, 82.2 mg, 82.3 mg. 82.4 mg, 82.5 mg, 82.6 mg,
82.7 mg, 82.8 mg, 82.9 mg, 83.0 mg, 83.1 mg, 83.2 mg, 83.3 mg, 83.4
mg, 83.5 mg, 83.6 mg, 83.7 mg, 83.8 mg, 83.9 mg, 84.0 mg, 84.1 mg,
84.2 mg, 84.3 mg. 84.4 mg, 84.5 mg, 84.6 mg, 84.7 mg, 84.8 mg, 84.9
mg, and 85.0 mg. In certain embodiments, the oligomeric compound is
present in the dosage unit at an amount selected from about 75.0
mg, about 75.1 mg, about 75.2 mg, about 75.3 mg, about 75.4 mg,
about 75.5 mg, about 75.6 mg, about 75.7 mg, about 75.8 mg, about
75.9 mg, about 76.0 mg, about 76.1 mg, about 76.2 mg, about 76.3
mg. about 76.4 mg, about 76.5 mg, about 76.6 mg, about 76.6 about
76.7 mg, about 76.8 mg, about 76.9 mg, about 77.0 mg, about 77.1
mg, about 77.2 mg, about 77.3 mg, about 77.4 mg, about 77.5 mg,
about 77.6 mg, about 77.7 mg, about 77.8 mg, about 77.9 mg, about
78.0 mg, about 78.1 mg, about 78.2 mg, about 78.3 mg. about 78.4
mg, about 78.5 mg, about 78.6 mg, about 78.7 mg, about 78.8 mg,
about 78.9 mg, about 79.0 mg, about 79.1 mg, about 79.2 mg, about
79.3 mg, about 79.4 mg, about 79.5 mg, about 79.6 mg, about 79.7
mg, about 79.8 mg, about 79.9 mg, about 80.0 mg, about 80.1 mg,
about 80.2 mg, about 80.3 mg. about 80.4 mg, about 80.5 mg, about
80.6 mg, about 80.7 mg, about 80.8 mg, about 80.9 mg, about 81.0
mg, about 81.1 mg, about 81.2 mg, about 81.3 mg, about 81.4 mg,
about 81.5 mg, about 81.6 mg, about 81.7 mg, about 81.8 mg, about
81.9 mg, about 82.0 mg, about 82.1 mg, about 82.2 mg, about 82.3
mg. about 82.4 mg, about 82.5 mg, about 82.6 mg, about 82.7 mg,
about 82.8 mg, about 82.9 mg, about 83.0 mg, about 83.1 mg, about
83.2 mg, about 83.3 mg, about 83.4 mg, about 83.5 mg, about 83.6
mg, about 83.7 mg, about 83.8 mg, about 83.9 mg, about 84.0 mg,
about 84.1 mg, about 84.2 mg, about 84.3 mg. about 84.4 mg, about
84.5 mg, about 84.6 mg, about 84.7 mg, about 84.8 mg, about 84.9
mg, and about 85.0 mg.
[0318] In certain embodiments, the oligomeric compound is present
in the dosage unit at an amount that falls within a range. In
certain embodiments, the range is selected from 10 mg to 140 mg,
from 10 mg to 130 mg, from 10 mg to 120 mg, from 10 mg to 110 mg,
from 10 mg to 100 mg, from 10 mg to 90 mg, from 10 mg to 80 mg,
from 10 mg to 70 mg, from 10 mg to 60 mg, from 10 mg to 50 mg, from
10 mg to 40 mg, from 10 mg to 30 mg, from 10 mg to 20 mg, from 20
mg to 140 mg, from 20 mg to 130 mg, from 20 mg to 120 mg, from 20
mg to 110 mg, from 20 mg to 100 mg, from 20 mg to 90 mg, from 20 mg
to 80 mg, from 20 mg to 70 mg, from 20 mg to 60 mg, from 20 mg to
50 mg, from 20 mg to 40 mg, from 20 mg to 30 mg, from 30 mg to 140
mg, from 30 mg to 130 mg, from 30 mg to 120 mg, from 30 mg to 110
mg, from 30 mg to 100 mg, from 30 mg to 90 mg, from 30 mg to 80 mg,
from 30 mg to 70 mg, from 30 mg to 60 mg, from 30 mg to 50 mg, from
30 mg to 40 mg, from 40 mg to 140 mg, from 40 mg to 130 mg, from 40
mg to 120 mg, from 40 mg to 110 mg, from 40 mg to 100 mg, from 40
mg to 90 mg, from 40 mg to 80 mg, from 40 mg to 70 mg, from 40 mg
to 60 mg, from 40 mg to 50 mg, from 50 mg to 140 mg, from 50 mg to
130 mg, from 50 mg to 120 mg, from 50 mg to 110 mg, from 50 mg to
100 mg, from 50 mg to 90 mg, from 50 mg to 80 mg, from 50 mg to 70
mg, from 50 mg to 60 mg, from 60 mg to 140 mg, from 60 mg to 130
mg, from 60 mg to 120 mg, from 60 mg to 110 mg, from 60 mg to 100
mg, from 60 mg to 90 mg, from 60 mg to 80 mg, from 60 mg to 70 mg,
from 70 mg to 140 mg, from 70 mg to 130 mg, from 70 mg to 120 mg,
from 70 mg to 110 mg, from 70 mg to 100 mg, from 70 mg to 90 mg,
from 70 mg to 80 mg, from 80 mg to 140 mg, from 80 mg to 130 mg,
from 80 mg to 120 mg, from 80 mg to 110 mg, from 80 mg to 100 mg,
from 80 mg to 90 mg, from 90 mg to 140 mg, from 90 mg to 130 mg,
from 90 mg to 120 mg, from 90 mg to 110 mg, from 90 mg to 100 mg,
from 100 mg to 140 mg, from 100 mg to 130 mg, from 100 mg to 120
mg, from 100 mg to 110 mg, from 110 mg to 140 mg, from 110 mg to
130 mg, from 110 mg to 120 mg, from 120 mg to 140 mg, from 120 mg
to 130 mg, from 130 mg to 140 mg, from 65 mg to 95 mg, from 65 mg
to 90 mg, from 65 mg to 85 mg from 65 mg to 80 mg, from 65 mg to 75
mg, from 65 mg to 70 mg, from 70 mg to 95 mg, from 70 mg to 85 mg,
from 70 mg to 75 mg, from 75 mg to 100 mg, from 75 mg to 95 mg,
from 75 mg to 90 mg, from 75 mg to 85 mg, from 75 mg to 80 mg, from
80 mg to 95 mg, from 80 mg to 85 mg, from 85 mg to 100 mg, from 85
mg to 90 mg, from 90 mg to 95 mg, from 95 mg to 100 mg, from 80 mg
to 89 mg, from 80 mg to 88 mg, from 80 mg to 87 mg, from 80 mg to
86 mg, from 80 mg to 84 mg, from 80 mg to 83 mg, from 80 mg to 82
mg, from 80 mg to 81 mg, from 81 mg to 90 mg, from 82 mg to 89 mg,
from 82 mg to 88 mg, from 82 mg to 87 mg, from 82 mg to 86 mg, from
82 mg to 85 mg, from 82 mg to 84 mg, from 82 mg to 83 mg, from 83
mg to 90 mg, from 83 mg to 89 mg, from 83 mg to 88 mg, from 83 mg
to 87 mg, from 83 mg to 86 mg, from 83 mg to 85 mg, from 83 mg to
84 mg, from 84 mg to 90 mg, from 84 mg to 89 mg, from 84 mg to 88
mg, from 84 mg to 87 mg, from 84 mg to 86 mg, from 84 mg to 85 mg,
from 85 mg to 89 mg, from 85 mg to 88 mg, from 85 mg to 87 mg, from
85 mg to 86 mg, from 86 mg to 90 mg, from 86 mg to 89 mg, from 86
mg to 88 mg, from 86 mg to 87 mg, from 87 mg to 90 mg, from 87 mg
to 89 mg, from 87 mg to 88 mg, from 88 mg to 90 mg, from 88 mg to
89 mg, and from 89 mg to 90 mg.
[0319] In certain embodiments, the oligomeric compound is present
in the dosage unit at an amount that is less than about 300 mg,
less than about 295 mg, less than about 290 mg, less than about 285
mg, less than about 280 mg, less than about 275 mg, less than about
270 mg, less than about 265 mg, less than about 260 mg, less than
about 255 mg, less than about 250 mg, less than about 245 mg, less
than about 240 mg, less than about 235 mg, less than about 230 mg,
less than about 225 mg, less than about 220 mg, less than about 215
mg, less than about 210 mg, less than about 205 mg, less than about
200 mg, less than about 195 mg, less than about 190 mg, less than
about 185 mg, less than about 180 mg, less than about 175 mg, less
than about 170 mg, less than about 165 mg, less than about 160 mg,
less than about 150 mg, less than about 145 mg, less than about 140
mg, less than about 135 mg, less than about 130 mg, less than about
125 mg, less than about 120 mg, less than about 115 mg, less than
about 110 mg, less than about 105 mg, less than about 100 mg, less
than about 95 mg, less than about 90 mg, less than about 85 mg,
less than about 80 mg, less than about 75 mg, less than about 70
mg, less than about 65 mg, less than about 60 mg, less than about
55 mg, less than about 50 mg, less than about 45 mg, less than
about 40 mg, less than about 35 mg, less than about 30 mg, less
than about 25 mg, or less than about 20 mg of an oligomeric
compound disclosed herein. The dosage unit may contain more than
about 5 mg or more than about 10 mg of the oligomeric compound.
[0320] In certain embodiments, the oligomeric compound is present
in the dosage unit at an amount that is less than 300 mg, less than
295 mg, less than 290 mg, less than 285 mg, less than 280 mg, less
than 275 mg, less than 270 mg, less than 265 mg, less than 260 mg,
less than 255 mg, less than 250 mg, less than 245 mg, less than 240
mg, less than 235 mg, less than 230 mg, less than 225 mg, less than
220 mg, less than 215 mg, less than 210 mg, less than 205 mg, less
than 200 mg, less than 195 mg, less than 190 mg, less than 185 mg,
less than 180 mg, less than 175 mg, less than 170 mg, less than 165
mg, less than 160 mg, less than 150 mg, less than 145 mg, less than
140 mg, less than 135 mg, less than 130 mg, less than 125 mg, less
than 120 mg, less than 115 mg, less than 110 mg, less than 105 mg,
less than 100 mg, less than 95 mg, less than 90 mg, less than 85
mg, less than 80 mg, less than 75 mg, less than 70 mg, less than 65
mg, less than 60 mg, less than 55 mg, less than 50 mg, less than 45
mg, less than 40 mg, less than 35 mg, less than 30 mg, less than 25
mg, or less than 20 mg of an oligomeric compound disclosed herein.
The dosage unit may contain more than 5 mg or more than 10 mg of
the oligomeric compound.
[0321] In certain embodiments, the oligomeric compound is present
in the dosage unit at an amount that is at least about 10 mg, at
least about 15 mg, at least about 20 mg, at least about 25 mg, at
least about 30 mg, at least about 35 mg, at least about 40 mg, at
least about 45 mg, at least about 50 mg, at least about 55 mg, at
least about 60 mg, at least about 65 mg, at least about 70 mg, at
least about 75 mg, at least about 80 mg, at least about 85 mg, at
least about 90 mg, at least about 95 mg, at least about 100 mg, at
least about 105 mg, at least about 115 mg, at least about 120 mg,
at least about 125 mg, at least about 130 mg, at least about 135
mg, at least about 140 mg, at least about 145 mg, or at least about
150 mg of an oligomeric compound disclosed herein.
[0322] In certain embodiments, the oligomeric compound is present
in the dosage unit at an amount that is at least 10 mg, at least 15
mg, at least 20 mg, at least 25 mg, at least 30 mg, at least 35 mg,
at least 40 mg, at least 45 mg, at least 50 mg, at least 55 mg, at
least 60 mg, at least 65 mg, at least 70 mg, at least 75 mg, at
least 80 mg, at least 85 mg, at least 90 mg, at least 95 mg, at
least about 100 mg, at least 105 mg, at least 115 mg, at least 120
mg, at least 125 mg, at least 130 mg, at least 135 mg, at least 140
mg, at least 145 mg, or at least 150 mg of an oligomeric compound
disclosed herein
[0323] In certain embodiments, pharmaceutical compositions
disclosed herein are provided as a volume of a solution comprising
an oligomeric compound and a pharmaceutically acceptable carrier or
diluent. In certain embodiments, the solution consists or consists
essentially of an oligomeric compound disclosed herein and a
pharmaceutically acceptable carrier or diluent. The volume may be
provided in a suitable container, such as a vial or syringe. Since
pharmaceutical compositions disclosed herein may be amenable to
self-administration, the syringe may be a pre-filled syringe, an
auto-injector syringe, or a combination thereof. For example, a
dosage unit described herein may be provided as a fixed volume in a
syringe for convenient administration. In certain embodiments, the
volume of the solution is 0.1 ml, 0.2 ml, 0.3 ml, 0.4 ml, 0.5 ml,
0.6 ml, 0.7 ml, 0.8 ml, 0.9 ml, 1.0 ml, 1.1 ml, 1.2 ml, 1.3 ml, 1.4
ml, 1.5 ml, 1.6 ml, 1.7 ml, 1.8 ml, 1.9 ml, or 2.0 ml. In certain
embodiments, the volume of the solution is about 0.1 ml, about 0.2
ml, about 0.3 ml, about 0.4 ml, about 0.5 ml, about 0.6 ml, about
0.7 ml, about 0.8 ml, about 0.9 ml, about 1.0 ml, about 1.1 ml,
about 1.2 ml, about 1.3 ml, about 1.4 ml, about 1.5 ml, about 1.6
ml, about 1.7 ml, about 1.8 ml, about 1.9 ml, or about 2.0 ml. In
certain embodiments, the volume of the solution is less than 1.0
ml, less than 1.5 ml, or 2.0 ml. In certain embodiments, the volume
of the solution is less than 1.0 ml. A volume of less than 2.0 ml
may be useful to reduce or avoid injection pain, adverse events at
the injection site, and injection site leakage.
[0324] In certain embodiments, pharmaceutical compositions
disclosed herein are provided as a volume of a solution comprising
an oligomeric compound and a pharmaceutically acceptable carrier or
diluent, wherein the volume of the solution is 0.1 ml to 1.5 ml,
0.1 ml to 1.4 ml, 0.1 ml to 1.3 ml, 0.1 ml to 1.2 ml, 0.1 ml to 1.1
ml, 0.1 ml to 1.0 ml, 0.1 ml to 0.9 ml, 0.1 ml to 0.8 ml, 0.1 ml to
0.7 ml, 0.1 ml to 0.6 ml, 0.1 ml to 0.5 ml, 0.1 ml to 0.4 ml, 0.1
ml to 0.3 ml, 0.1 ml to 0.2 ml, 0.2 ml to 1.5 ml, 0.2 ml to 1.4 ml,
0.2 ml to 1.3 ml, 0.2 ml to 1.2 ml, 0.2 ml to 1.1 ml, 0.2 ml to 1.0
ml, 0.2 ml to 0.9 ml, 0.2 ml to 0.8 ml, 0.2 ml to 0.7 ml, 0.2 ml to
0.6 ml, 0.2 ml to 0.5 ml, 0.2 ml to 0.4 ml, 0.2 ml to 0.3 ml, 0.3
ml to 1.5 ml, 0.3 ml to 1.4 ml, 0.3 ml to 1.3 ml, 0.3 ml to 1.2 ml,
0.3 ml to 1.1 ml, 0.3 ml to 1.0 ml, 0.3 ml to 0.9 ml, 0.3 ml to 0.8
ml, 0.3 ml to 0.7 ml, 0.3 ml to 0.6 ml, 0.3 ml to 0.5 ml, 0.3 ml to
0.4 ml, 0.4 ml to 1.5 ml, 0.4 ml to 1.4 ml, 0.4 ml to 1.3 ml, 0.4
ml to 1.2 ml, 0.4 ml to 1.1 ml, 0.4 ml to 1.0 ml, 0.4 ml to 0.9 ml,
0.4 ml to 0.8 ml, 0.4 ml to 0.7 ml, 0.4 ml to 0.6 ml, 0.4 ml to 0.5
ml, 0.5 ml to 1.5 ml, 0.5 ml to 1.4 ml, 0.5 ml to 1.3 ml, 0.5. ml
to 1.2 ml, 0.5 ml to 1.1 ml, 0.5 ml to 1.0 ml, 0.5 ml to 0.9 ml,
0.5 ml to 0.8 ml, 0.5 ml to 0.7 ml, 0.5 ml to 0.6 ml, 0.6 ml to 1.5
ml, 0.6 ml to 1.4 ml, 0.6 mo to 1.3 ml, 0.6 ml to 1.2 ml, 0.6 ml to
1.1 ml, 0.6 ml to 1.0 ml, 0.6 ml to 0.9 ml, 0.6 ml to 0.8 ml, 0.6
ml to 0.7 ml, 0.7 ml, to 1.5 ml, 0.7 ml to 1.4 ml, 0.7 ml to 1.3
ml, 0.7 ml to 1.2 ml, 0.7 ml to 1.1 ml, 0.7 ml to 1.0 ml, 0.7 ml to
0.9 ml, 0.7 ml to 0.8 ml, 0.8 ml to 1.5 ml, 0.8 ml to 1.4 ml, 0.8
ml to 1.3 ml, 0.8 ml to 1.2 ml, 0.8 ml to 1.1 ml, 0.8 ml to 1.0 ml.
0.8 ml to 0.9 ml, 0.9 ml, to 1.5 ml, 0.9 ml to 1.4 ml, 0.9 ml to
1.3 ml, 0.9 ml to 1.2 ml, 0.9 ml, to 1.1 ml, 0.9 ml to 1.0 ml, 1.0
ml to 1.5 ml, 1.0 ml to 1.4 ml, 1.0 ml to 1.3 ml, 1.0 ml to 1.2 ml,
1.0 ml to 1.1 ml, 1.1 ml to 1.5 ml, 1.1 ml to 1.4 ml, 1.1 ml to 1.3
ml, 1.1 ml to 1.2 ml, 1.2 ml to 1.5 ml, 1.2 ml to 1.4 ml, 1.2 ml to
1.3 ml, 1.3 ml to 1.5 ml, 1.3 ml to 1.4 ml, 1.4 ml to, and 1.5
ml.
[0325] In certain embodiments, pharmaceutical compositions comprise
a solution of an oligomeric compound disclosed herein in a
pharmaceutically acceptable carrier or diluent at a concentration
of 5 mg/ml, 10 mg/ml, 15 mg/ml, 20 mg/ml, 25 mg/ml, 30 mg/ml, 35
mg/ml, 40 mg/ml, 45 mg/ml, 50 mg/ml, 55 mg/ml, 60 mg/ml, 65 mg/ml,
70 mg/ml, 75 mg/ml, 80 mg/ml, 85 mg/ml, 90 mg/ml, 95 mg/ml, 100
mg/ml, 105 mg/ml, 110 mg/ml, 115 mg/ml, 120 mg/ml, 125 mg/ml, 130
mg/ml, 135 mg/ml, 140 mg/ml, 145 mg/ml, 150 mg/ml, 155 mg/ml, or
160 mg/ml. In certain embodiments, pharmaceutical compositions
comprise a solution of an oligomeric compound disclosed herein in a
pharmaceutically acceptable carrier or diluent at a concentration
of about 5 mg/ml, about 10 mg/ml, about 15 mg/ml, about 20 mg/ml,
about 25 mg/ml, about 30 mg/ml, about 35 mg/ml, about 40 mg/ml,
about 45 mg/ml, about 50 mg/ml, about 55 mg/ml, about 60 mg/ml,
about 65 mg/ml, about 70 mg/ml, about 75 mg/ml, about 80 mg/ml,
about 85 mg/ml, about 90 mg/ml, about 95 mg/ml, about 100 mg/ml,
about 105 mg/ml, about 110 mg/ml, about 115 mg/ml, about 120 mg/ml,
about 125 mg/ml, about 130 mg/ml, about 135 mg/ml, about 140 mg/ml,
about 145 mg/ml, about 150 mg/ml, about 155 mg/ml, or about 160
mg/ml.
[0326] In certain embodiments, pharmaceutical compositions comprise
a solution of an oligomeric compound disclosed herein in a
pharmaceutically acceptable carrier or diluent at a concentration
of 20 mg/ml, 21 mg/ml, 22 mg/ml, 23 mg/ml, 24 mg/ml, 25 mg/ml, 26
mg/ml, 27 mg/ml, 28 mg/ml, 29 mg/ml, 30 mg/ml, 31 mg/ml, 32 mg/ml,
33 mg/ml, 34 mg/ml, 35 mg/ml, 36 mg/ml, 37 mg/ml, 38 mg/ml, 39
mg/ml, 40 mg/ml, 41 mg/ml, 42 mg/ml, 43 mg/ml, 44 mg/ml, 45 mg/ml,
46 mg/ml, 47 mg/ml, 48 mg/ml, 49 mg/ml, 50 mg/ml, 51 mg/ml, 52
mg/ml, 53 mg/ml, 54 mg/ml, 55 mg/ml, 56 mg/ml, 57 mg/ml, 58 mg/ml,
59 mg/ml, 60 mg/ml, 61 mg/ml, 62 mg/ml, 63 mg/ml, 64 mg/ml, 65
mg/ml, 66 mg/ml, 67 mg/ml, 68 mg/ml, 69 mg/ml, 70 mg/ml, 71 mg/ml,
72 mg/ml, 73 mg/ml, 74 mg/ml, 75 mg/ml, 76 mg/ml, 77 mg/ml, 78
mg/ml, 79 mg/ml, 80 mg/ml, 81 mg/ml, 82 mg/ml, 83 mg/ml, 84 mg/ml,
85 mg/ml, 86 mg/ml, 87 mg/ml, 88 mg/ml, 89 mg/ml, 90 mg/ml, 91
mg/ml, 92 mg/ml, 93 mg/ml, 94 mg/ml, 95 mg/ml, 96 mg/ml, 97 mg/ml,
98 mg/ml, 99 mg/ml, 100 mg/ml, 101 mg/ml, 102 mg/ml, 103 mg/ml, 104
mg/ml, 105 mg/ml, 106 mg/ml, 107 mg/ml, 108 mg/ml, 109 mg/ml, 110
mg/ml, 111 mg/ml, 112 mg/ml, 113 mg/ml, 114 mg/ml, 115 mg/ml, 116
mg/ml, 117 mg/ml, 118 mg/ml, 119 mg/ml, 120 mg/ml, 121 mg/ml, 122
mg/ml, 123 mg/ml, 124 mg/ml, 125 mg/ml, 126 mg/ml, 127 mg/ml, 128
mg/ml, 129 mg/ml, 130 mg/ml, 131 mg/ml, 132 mg/ml, 133 mg/ml, 134
mg/ml, 135 mg/ml, 136 mg/ml, 137 mg/ml, 138 mg/ml, 139 mg/ml, and
140 mg/ml. In certain embodiments, the oligomeric compound is
present in the dosage unit at an amount selected from about 20
mg/ml, about 21 mg/ml, about 22 mg/ml, about 23 mg/ml, about 24
mg/ml, about 25 mg/ml, about 26 mg/ml, about 27 mg/ml, about 28
mg/ml, about 29 mg/ml, about 30 mg/ml, about 31 mg/ml, about 32
mg/ml, about 33 mg/ml, about 34 mg/ml, about 35 mg/ml, about 36
mg/ml, about 37 mg/ml, about 38 mg/ml, about 39 mg/ml, about 40
mg/ml, about 41 mg/ml, about 42 mg/ml, about 43 mg/ml, about 44
mg/ml, about 45 mg/ml, about 46 mg/ml, about 47 mg/ml, about 48
mg/ml, about 49 mg/ml, about 50 mg/ml, about 51 mg/ml, about 52
mg/ml, about 53 mg/ml, about 54 mg/ml, about 55 mg/ml, about 56
mg/ml, about 57 mg/ml, about 58 mg/ml, about 59 mg/ml, about 60
mg/ml, about 61 mg/ml, about 62 mg/ml, about 63 mg/ml, about 64
mg/ml, about 65 mg/ml, about 66 mg/ml, about 67 mg/ml, about 68
mg/ml, about 69 mg/ml, about 70 mg/ml, about 71 mg/ml, about 72
mg/ml, about 73 mg/ml, about 74 mg/ml, about 75 mg/ml, about 76
mg/ml, about 77 mg/ml, about 78 mg/ml, about 79 mg/ml, about 80
mg/ml, about 81 mg/ml, about 82 mg/ml, about 83 mg/ml, about 84
mg/ml, about 85 mg/ml, about 86 mg/ml, about 87 mg/ml, about 88
mg/ml, about 89 mg/ml, about 90 mg/ml, about 91 mg/ml, about 92
mg/ml, about 93 mg/ml, about 94 mg/ml, about 95 mg/ml, about 96
mg/ml, about 97 mg/ml, about 98 mg/ml, about 99 mg/ml, about 100
mg/ml, about 101 mg/ml, about 102 mg/ml, about 103 mg/ml, about 104
mg/ml, about 105 mg/ml, about 106 mg/ml, about 107 mg/ml, about 108
mg/ml, about 109 mg/ml, about 110 mg/ml, about 111 mg/ml, about 112
mg/ml, about 113 mg/ml, about 114 mg/ml, about 115 mg/ml, about 116
mg/ml, about 117 mg/ml, about 118 mg/ml, about 119 mg/ml, about 120
mg/ml, about 121 mg/ml, about 122 mg/ml, about 123 mg/ml, about 124
mg/ml, about 125 mg/ml, about 126 mg/ml, about 127 mg/ml, about 128
mg/ml, about 129 mg/ml, about 130 mg/ml, about 131 mg/ml, about 132
mg/ml, about 133 mg/ml, about 134 mg/ml, about 135 mg/ml, about 136
mg/ml, about 137 mg/ml, about 138 mg/ml, about 139 mg/ml, and about
140 mg/ml.
[0327] In certain embodiments, pharmaceutical compositions comprise
a solution of an oligomeric compound disclosed herein in a
pharmaceutically acceptable carrier or diluent at a concentration
of 20 mg/ml to 180 mg/ml, 20 mg/ml to 170 mg, 20 mg/ml to 160
mg/ml, 20 mg/ml to 150 mg/ml, 20 mg/ml to 140 mg/ml, 20 mg/ml to
130 mg/ml, 20 mg/ml to 120 mg/ml, 20 mg/ml to 110 mg/ml, 20 mg/ml
to 100 mg/ml, 20 mg/ml to 90 mg/ml, 20 mg/ml to 80 mg/ml, 20 mg/ml
to 70 mg/ml, 20 mg/ml to 60 mg/ml, 20 mg/ml to 50 mg/ml, 20 mg/ml
to 40 mg/ml, 20 mg/ml to 30 mg/ml, 30 mg/ml to 180 mg/ml, 30 mg/ml
to 170 mg, 30 mg/ml to 160 mg/ml, 30 mg/ml to 150 mg/ml, 30 mg/ml
to 140 mg/ml, 30 mg/ml to 130 mg/ml, 30 mg/ml to 120 mg/ml, 30
mg/ml to 110 mg/ml, 30 mg/ml to 100 mg/ml, 30 mg/ml to 90 mg/ml, 30
mg/ml to 80 mg/ml, 30 mg/ml to 70 mg/ml, 30 mg/ml to 60 mg/ml, 30
mg/ml to 50 mg/ml, 30 mg/ml to 40 mg/ml, 40 mg/ml to 180 mg/ml, 40
mg/ml to 170 mg, 40 mg/ml to 160 mg/ml, 40 mg/ml to 150 mg/ml, 40
mg/ml to 140 mg/ml, 40 mg/ml to 130 mg/ml, 40 mg/ml to 120 mg/ml,
40 mg/ml to 110 mg/ml, 40 mg/ml to 100 mg/ml, 40 mg/ml to 90 mg/ml,
40 mg/ml to 80 mg/ml, 40 mg/ml to 70 mg/ml, 40 mg/ml to 60 mg/ml,
40 mg/ml to 50 mg/ml, 50 mg/ml to 180 mg/ml, 50 mg/ml to 170 mg, 50
mg/ml to 160 mg/ml, 50 mg/ml to 150 mg/ml, 50 mg/ml to 140 mg/ml,
50 mg/ml to 130 mg/ml, 50 mg/ml to 120 mg/ml, 50 mg/ml to 110
mg/ml, 50 mg/ml to 100 mg/ml, 50 mg/ml to 90 mg/ml, 50 mg/ml to 80
mg/ml, 50 mg/ml to 70 mg/ml, 50 mg/ml to 60 mg/ml, 60 mg/ml to 180
mg/ml, 60 mg/ml to 170 mg, 60 mg/ml to 160 mg/ml, 60 mg/ml to 150
mg/ml, 60 mg/ml to 140 mg/ml, 60 mg/ml to 130 mg/ml, 60 mg/ml to
120 mg/ml, 60 mg/ml to 110 mg/ml, 60 mg/ml to 100 mg/ml, 60 mg/ml
to 90 mg/ml, 60 mg/ml to 80 mg/ml, 60 mg/ml to 70 mg/ml, 70 mg/ml
to 180 mg/ml, 70 mg/ml to 170 mg, 70 mg/ml to 160 mg/ml, 70 mg/ml
to 150 mg/ml, 70 mg/ml to 140 mg/ml, 70 mg/ml to 130 mg/ml, 70
mg/ml to 120 mg/ml, 70 mg/ml to 110 mg/ml, 70 mg/ml to 100 mg/ml,
70 mg/ml to 90 mg/ml, 70 mg/ml to 80 mg/ml, 80 mg/ml to 180 mg/ml,
80 mg/ml to 170 mg, 80 mg/ml to 160 mg/ml, 80 mg/ml to 150 mg/ml,
80 mg/ml to 140 mg/ml, 80 mg/ml to 130 mg/ml, 80 mg/ml to 120
mg/ml, 80 mg/ml to 110 mg/ml, 80 mg/ml to 100 mg/ml, 80 mg/ml to 90
mg/ml, 90 mg/ml to 180 mg/ml, 90 mg/ml to 170 mg, 90 mg/ml to 160
mg/ml, 90 mg/ml to 150 mg/ml, 90 mg/ml to 140 mg/ml, 90 mg/ml to
130 mg/ml, 90 mg/ml to 120 mg/ml, 90 mg/ml to 110 mg/ml, 90 mg/ml
to 100 mg/ml, 100 mg/ml to 180 mg/ml, 100 mg/ml to 170 mg, 100
mg/ml to 160 mg/ml, 100 mg/ml to 150 mg/ml, 100 mg/ml to 140 mg/ml,
100 mg/ml to 130 mg/ml, 100 mg/ml to 120 mg/ml, 100 mg/ml to 110
mg/ml, 110 mg/ml to 180 mg/ml, 110 mg/ml to 170 mg, 110 mg/ml to
160 mg/ml, 110 mg/ml to 150 mg/ml, 110 mg/ml to 140 mg/ml, 110
mg/ml to 130 mg/ml, 110 mg/ml to 120 mg/ml, 120 mg/ml to 180 mg/ml,
120 mg/ml to 170 mg, 120 mg/ml to 160 mg/ml, 120 mg/ml to 150
mg/ml, 120 mg/ml to 140 mg/ml, 120 mg/ml to 130 mg/ml, 130 mg/ml to
180 mg/ml, 130 mg/ml to 170 mg, 130 mg/ml to 160 mg/ml, 130 mg/ml
to 150 mg/ml, 130 mg/ml to 140 mg/ml, 140 mg/ml to 180 mg/ml, 140
mg/ml to 170 mg/ml, 140 mg/ml to 160 mg/ml, 140 mg/ml to 150 mg/ml,
150 mg/ml to 180 mg/ml, 150 mg/ml to 170 mg/ml, 150 mg/ml to 160
mg/ml, 160 mg/ml to 180 mg/ml, 160 mg/ml to 170 mg/ml, or 170 mg/ml
to 180 mg/ml.
[0328] In certain embodiments, pharmaceutical compositions
disclosed herein comprise a sterile lyophilized oligomeric compound
that can be reconstituted with a suitable diluent for injection. In
certain embodiments, pharmaceutical compositions disclosed herein
consist or consist essentially of a sterile lyophilized oligomeric
compound that can be reconstituted with a suitable diluent for
injection. In certain embodiments, the reconstituted product is
administered as a subcutaneous injection after dilution. In certain
embodiments, a sterile lyophilized oligomeric compound may consist
of the oligomeric compound which has been prepared in distilled
water for injection, adjusted to pH 7.0-9.0 with acid or base
during preparation, and then lyophilized. The sterile lyophilized
oligomeric compound may be packaged in a Type I, clear glass vial
(ammonium sulfate-treated), stoppered with a bromobutyl rubber
closure and sealed with an aluminum FLIP-OFF.RTM. overseal. In
certain embodiments, the sterile lyophilized oligomeric compound
comprises Compound No. 957943. In certain embodiments, the sterile
lyophilized oligomeric compound consists or consists essentially of
Compound No. 957943.
[0329] VIII. Certain Dosing Regimens
[0330] Pharmaceutical compositions disclosed herein may be suitable
for acute treatment, temporary treatment, ongoing (prophylactic)
treatment, chronic treatment, or a combination thereof. In certain
embodiments, methods comprise continually administering a
pharmaceutical composition disclosed herein as a prophylactic
measure. In certain embodiments, methods comprise temporarily
administering a pharmaceutical composition disclosed herein. For
example, methods may comprise administering a pharmaceutical
composition disclosed herein to an animal within 24 hours of the
animal experiencing a thromboembolic event. In certain embodiments,
methods comprise administering a pharmaceutical composition
disclosed herein to an animal before surgery, during surgery, after
surgery, or a combination thereof. In certain embodiments, methods
comprise prophylactically administering a pharmaceutical
composition disclosed herein on a monthly basis to an animal with a
FXI associated disease or condition who is at risk for a thrombotic
event. In certain embodiments, the pharmaceutical composition
comprises Compound No. 957943. In certain embodiments, the
pharmaceutical composition consists or consists essentially of
Compound No. 957943 and a pharmaceutically acceptable carrier or
diluent.
[0331] In certain embodiments, pharmaceutical compositions
disclosed herein are useful for treating an animal with a FXI
associated disease or condition. In certain embodiments, methods
comprise administering a pharmaceutical composition disclosed
herein to an animal only once. In certain embodiments, methods
comprise administering a pharmaceutical composition disclosed
herein to an animal at least twice. In certain embodiments, methods
comprise administering a pharmaceutical composition disclosed
herein at least 3, at least 4, at least 5, at least 6, at least 8,
or at least 10 times. In certain embodiments, methods comprise
administering a pharmaceutical composition disclosed herein less
than 20 times, less than 15 times, less than 10 times, or less than
5 times.
[0332] In certain embodiments, methods comprise administering to an
animal a first dose and a second dose of an oligomeric compound
disclosed herein. In certain embodiments, the first dose and the
second dose are the same. In certain embodiments, the first dose
and the second dose are different. In certain embodiments, the
first dose is greater than the second dose. In certain embodiments,
the second dose is greater than the first dose. In certain
embodiments, the first dose is at least 10%, at least 20%, at least
30%, at least 40%, at least 50%, at least 60%, at least 70%, at
least 80%, at least 90%, or at least 100% greater than the second
dose. In certain embodiments, the second dose is at least 10%, at
least 20%, at least 30%, at least 40%, at least 50%, at least 60%,
at least 70%, at least 80%, at least 90%, or at least 100% greater
than the first dose. In certain embodiments, the first dose and the
second dose are separated by 5, 10, 15, 20, 25, 30, 35, or 40 days.
In certain embodiments, the first dose and the second dose are
separated by 1, 2, 3, 4, 5, or 6 months. In certain embodiments,
the first dose and the second dose are separated by 20, 21, 22, 23,
24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or
40 days. In certain embodiments, the first dose and the second dose
are separated by 20 to 40 days, 22 to 38 days, 24 to 36 days, 26 to
34 days, 27 to 32 days, 28 to 32 days, or 29 to 31 days. In certain
embodiments, the first dose and the second dose are separated by 1,
2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks. In certain embodiments, the
first dose and the second dose are separated by 1 to 2 days, 2 to 4
days, 3 to 5 days, 4 to 6 days, 5 to 7 days, 6 to 8 days, 7 to 9
days, 8 to 10 days, 9 to 11 days, 10 to 12 days, 11 to 13 days, 12
to 14 days, 13 to 15 days, 14 to 16 days, 15 to 17 days, 16 to 18
days, 17 to 19 days, or 18 to 20 days.
[0333] In certain embodiments, methods comprise administering a
pharmaceutical composition disclosed herein monthly or about
monthly. In certain embodiments, methods comprise administering a
composition disclosed herein to an animal monthly or about monthly
for at least two months, at least three months, at least four
months, at least five months, or at least six months. In certain
embodiments, methods comprise administering a pharmaceutical
composition disclosed herein weekly or about weekly. In certain
embodiments, methods comprise administering a composition disclosed
herein to an animal weekly or about weekly for less than 2 weeks,
less than 3 weeks, less than 4 weeks, less than 5 weeks, less than
6 weeks, less than 8 week, less than 12 weeks, less than 16 weeks,
or less than 20 weeks.
[0334] In certain embodiments, methods comprise administering an
oligomeric compound according to a first dosing regimen and
subsequently administering the oligomeric compound according to a
second dosing regimen. In certain embodiments, the first dosing
regimen comprises administering the oligomeric compound at a first
dose and at a first frequency and the second dosing regimen
comprises administering the oligomeric compound at a second dose
and at a second frequency. In certain embodiments, the first
frequency is greater than the second frequency. By way of
non-limiting example, the first frequency may be greater than once
monthly (2 times, 3 times or 4 times per month) and the second
frequency may be once monthly. In certain embodiments, the first
frequency is less than the second frequency. In certain
embodiments, the first dose is greater than the second dose and the
first frequency is greater than the second frequency. In certain
embodiments, the first dose is less than the second dose and the
first frequency is greater than the second frequency. In certain
embodiments, the first dose is greater than the second dose and the
first frequency is less than the second frequency. In certain
embodiments, the first dose is less than the second dose and the
first frequency is less than the second frequency. In certain
embodiments, the first dose and the second dose are the same, and
the first frequency is greater than the second frequency. In
certain embodiments, the first dose and the second dose are the
same and the first frequency is less than the second frequency. In
certain embodiments, the first dose is greater than the second
dose, and the first frequency and the second frequency are the
same. In certain embodiments, the first dose is less than the
second dose and the first frequency and the second frequency are
the same. In certain embodiments, at least one of the first
frequency and the second frequency are selected from about every 5,
about every 10, about every 15, about every 20, about every 25,
about every 30, about every 35, or about every 40 days. In certain
embodiments, at least one of the first frequency and the second
frequency is monthly. In certain embodiments, at least one of the
first frequency and the second frequency is weekly. In certain
embodiments, the first dose is at least 10%, at least 20%, at least
30%, at least 40%, at least 50%, at least 60%, at least 70%, at
least 80%, at least 90%, or at least 100% greater than the second
dose. In certain embodiments, the second dose is at least 10%, at
least 20%, at least 30%, at least 40%, at least 50%, at least 60%,
at least 70%, at least 80%, at least 90%, or at least 100% greater
than the first dose.
[0335] In certain embodiments, methods comprise a human subject
self-administering a pharmaceutical composition disclosed herein.
In certain embodiments, methods disclosed herein comprise a human
subject self-administering a pharmaceutical composition disclosed
herein by subcutaneous injection. In certain embodiments, methods
disclosed herein comprise self-administering monthly. In general,
pharmaceutical compositions disclosed herein are prepared for
subcutaneous administration and a single dose can be provided with
a single injection, which makes these pharmaceutical compositions
amenable to self-administration. A single injection is possible
because oligomeric compounds disclosed herein are highly potent and
only a small amount needs to be dissolved in a small volume of a
carrier or diluent to provide a dosage unit. Due to the potency and
efficacy of oligomeric compounds disclosed herein, in certain
embodiments, the animal may only need to receive a dose on a
monthly basis to experience therapeutic effects. Such dosing
regimens that allow for self-administration on a monthly basis are
desirable to patients when compared to dosing regimens of other
therapies aimed to treat subjects with FXI associated conditions
and diseases, the latter of which may require in-clinic intravenous
injection and more frequent administration.
[0336] In certain embodiments, administration of a therapeutically
effective amount of a pharmaceutical composition as described
herein is accompanied by monitoring an amount of a FXI RNA, an
amount of a FXI protein, and/or FXI activity in the plasma or serum
of an animal, to determine the animal's response to administration
of the pharmaceutical composition. An animal's response to
administration of the pharmaceutical composition may be used by a
physician to determine the amount and duration of therapeutic
intervention.
[0337] IX. Certain Combination Therapies
[0338] In certain embodiments, methods comprise co-administering
one or more pharmaceutical compositions described herein with one
or more other pharmaceutical agents. In certain embodiments, such
one or more other pharmaceutical agents are designed to treat the
same disease or condition as the one or more pharmaceutical
compositions described herein. In certain embodiments, such one or
more other pharmaceutical agents are designed to treat a different
disease or condition as the one or more pharmaceutical compositions
described herein. In certain embodiments, such one or more other
pharmaceutical agents are designed to treat an undesired side
effect of one or more pharmaceutical compositions described herein.
In certain embodiments, methods comprise co-administering one or
more pharmaceutical compositions described herein with one or more
other pharmaceutical agents to treat an undesired effect of the
other pharmaceutical composition. In certain embodiments, methods
comprise co-administering one or more pharmaceutical compositions
described herein with one or more other pharmaceutical agents to
produce a combinational effect. In certain embodiments, methods
comprise co-administering one or more pharmaceutical compositions
described herein with one or more other pharmaceutical agents to
produce a synergistic effect. In certain embodiments, the
pharmaceutical composition comprises Compound No. 957943. In
certain embodiments, the pharmaceutical composition consists or
consists essentially of Compound No. 957943 and a pharmaceutically
acceptable carrier or diluent.
[0339] In certain embodiments, methods comprise co-administering
one or more pharmaceutical compositions described herein with one
or more other pharmaceutical agents at the same time. In certain
embodiments, methods comprise co-administering one or more
pharmaceutical compositions described herein with one or more other
pharmaceutical agents at different times. In certain embodiments,
one or more pharmaceutical compositions described herein and one or
more other pharmaceutical agents are prepared together in a single
formulation. In certain embodiments, one or more pharmaceutical
compositions described herein and one or more other pharmaceutical
agents are prepared separately.
[0340] In certain embodiments, pharmaceutical agents that may be
co-administered with a pharmaceutical composition described herein
include anticoagulant or antiplatelet agents. In certain
embodiments, pharmaceutical agents that may be co-administered with
a pharmaceutical composition described herein include
NSAID/Cyclooxygenase inhibitors, such as, aspirin. In certain
embodiments, pharmaceutical agents that may be co-administered with
a pharmaceutical composition described herein include adenosine
diphosphate (ADP) receptor inhibitors, such as, clopidogrel
(PLAVIX) and ticlopidine (TICLID). In certain embodiments,
pharmaceutical agents that may be co-administered with a
pharmaceutical composition described herein include
phosphodiesterase inhibitors, such as, cilostazol (PLETAL). In
certain embodiments, pharmaceutical agents that may be
co-administered with a pharmaceutical composition described herein
include, glycoprotein IIB/IIIA inhibitors, such as, abciximab
(REOPRO), eptifibatide (INTEGRILIN), tirofiban (AGGRASTAT), and
defibrotide. In certain embodiments, pharmaceutical agents that may
be co-administered with a pharmaceutical composition described
herein include, adenosine reuptake inhibitors, such as,
dipyridamole (PERSANTINE). In certain embodiments, pharmaceutical
agents that may be co-administered with a pharmaceutical
composition described herein include, but are not limited to,
warfarin (and related coumarins), heparin, direct thrombin
inhibitors (such as lepirudin, bivalirudin), apixaban, LOVENOX
(enoxaparin), and small molecular compounds that interfere directly
with the enzymatic action of particular coagulation factors (e.g.
rivaroxaban, which interferes with Factor Xa). In certain
embodiments, the anticoagulant or antiplatelet agent is
administered prior to administration of a pharmaceutical
composition described herein. In certain embodiments, the
anticoagulant or antiplatelet agent is administered following
administration of a pharmaceutical composition described herein. In
certain embodiments the anticoagulant or antiplatelet agent is
administered at the same time as a pharmaceutical composition
described herein. In certain embodiments, the dosage unit of a
co-administered anticoagulant or antiplatelet agent is the same as
the dosage unit that would be administered if the anticoagulant or
antiplatelet agent was administered alone. In certain embodiments
the dosage unit of a co-administered anticoagulant or antiplatelet
agent is lower than the dosage unit that would be administered if
the anticoagulant or antiplatelet agent was administered alone. In
certain embodiments the dosage unit of a co-administered
anticoagulant or antiplatelet agent is greater than the dosage unit
that would be administered if the anticoagulant or antiplatelet
agent was administered alone.
[0341] In certain embodiments, methods may comprise
co-administering a pharmaceutical composition described herein with
an anti-inflammatory agent. In certain embodiments, the
anti-inflammatory agent modifies a symptom of the inflammatory
disease or condition. In certain embodiments, the anti-inflammatory
agent modifies progression of the inflammatory disease or
condition. Non-limiting examples of inflammatory diseases and
conditions are autoimmune diseases (e.g. arthritis, colitis or
diabetes), trauma, surgery, sepsis, allergic inflammation, and
asthma. Non-limiting examples of anti-inflammatory agents include,
but are not limited to, methotrexate, abatacept, infliximab,
cyclophosphamide, azathioprine, corticosteroids, cyclosporin A,
aminosalicylates, sulfasalazine, hydroxychloroquine, leflunomide,
etanercept, efalizumab, 6-mercapto-purine (6-MP), and tumor
necrosis factor-alpha (TNFalpha), and other cytokine blockers or
antagonists. In certain embodiments, the anti-inflammatory agent
comprises a non-steroidal anti-inflammatory drug (NSAID). In
certain embodiments, NSAIDs reduce inflammatory reactions in an
animal. NSAIDS include, but are not limited to, acetyl salicylic
acid, choline magnesium salicylate, diflunisal, magnesium
salicylate, salsalate, sodium salicylate, diclofenac, etodolac,
fenoprofen, flurbiprofen, indomethacin, ketoprofen, ketorolac,
meclofenamate, naproxen, nabumetone, phenylbutazone, piroxicam,
sulindac, tolmetin, acetaminophen, ibuprofen, Cox-2 inhibitors,
meloxicam and tramadol. In certain embodiments, methods comprise
administering a pharmaceutical composition disclosed herein and an
anti-inflammatory agent concomitantly or sequentially. In certain
embodiments, methods comprise administering the anti-inflammatory
agent prior to administering the pharmaceutical composition
described herein. In certain embodiments, methods comprise
administering the anti-inflammatory agent after administering a
pharmaceutical composition described herein. In certain
embodiments, methods comprise administering the anti-inflammatory
agent and a pharmaceutical composition described herein at the same
time. In certain embodiments, the dosage unit of a co-administered
anti-inflammatory agent is the same as the dosage unit that would
be administered if the anti-inflammatory agent was administered
alone. In certain embodiments the dosage unit of a co-administered
anti-inflammatory agent is lower than the dosage unit that would be
administered if the anti-inflammatory agent was administered alone.
In certain embodiments the dosage unit of a co-administered
anti-inflammatory agent is greater than the dosage unit that would
be administered if the anticoagulant or antiplatelet agent was
administered alone.
[0342] In certain embodiments, the co-administration of a second
pharmaceutical agent enhances the anticoagulant or
anti-inflammatory effect of a pharmaceutical composition described
herein, such that co-administration of the two results in an
anticoagulant or anti-inflammatory effect that is greater than the
effect of administering the pharmaceutical composition described
herein. In certain embodiments, the co-administration results in
anticoagulant or anti-inflammatory effects that are additive of the
effects of the two when administered alone. In certain embodiments,
the co-administration results in anticoagulant or anti-inflammatory
effects that are supra-additive of the effects of the two when
administered alone. In certain embodiments, co-administration of a
second pharmaceutical agent increases an antithrombotic activity or
anti-inflammatory activity of the pharmaceutical composition
relative to that provided by the pharmaceutical composition alone,
without increased bleeding risk.
[0343] In certain embodiments, methods comprise co-administering a
pharmaceutical composition described herein and an antiplatelet
therapy to an animal in need thereof. In certain embodiments, the
animal in need thereof may be an animal having a condition selected
from thromboembolism, atrial fibrillation, a heart valve disorder,
valvular heart disease, stroke, coronary artery disease (CAD), and
a mechanical heart valve, and a combination thereof. In certain
embodiments, administering a pharmaceutical composition described
herein in combination with an antiplatelet therapy results in
little to no detectable increase in risk of bleeding as compared to
antiplatelet therapy alone. In certain embodiments, the risk
profile or risk indications are unchanged over antiplatelet therapy
alone.
[0344] In certain embodiments, methods comprise administering a
pharmaceutical composition described herein to a dialysis patient,
including but not limited to an animal with end stage renal disease
(ESRD) or chronic kidney disease (CKD), wherein the animal is
co-administered at least one pharmaceutical agent selected from
heparin, erythropoietin, darbopoetin, iron, Vitamin D or analogues
thereof, phosphate binders, and a combination thereof. In certain
embodimenst, the animal receives dialysis. In certain embodiments,
the pharmaceutical agent is administered at a dose that does not
differ from a comparative dose that would be prescribed in the
absence of administering the pharmaceutical composition.
[0345] X. Certain Indications
[0346] In certain embodiments, methods comprise administering a
pharmaceutical composition described herein to an animal with a
thromboembolic condition. In certain embodiments, methods comprise
administering a pharmaceutical composition described herein to an
animal at risk for a thromboembolic condition. Thromboembolic
conditions include, but are not limited to, thrombosis, embolism,
thromboembolism, infarct, deep vein thrombosis, pulmonary embolism,
myocardial infarction, stroke, and coronary artery disease (CAD).
Risk factors for developing a thromboembolic condition include, a
genetic, situational, disease, or environmental factor, or a
combination thereof. In certain embodiments, such factors may
include, but are not limited to, surgery, cancer, malignancy,
pregnancy, older age, use of oral contraceptives, immobility,
including travel-related immobility, sepsis, having a mechanical
heart valve, valvular heart disease, atrial fibrillation,
atherosclerosis atrial fibrillation, genetic predisposition,
antiphospholipid syndrome, and inherited or acquired prothrombotic
clotting disorders, such as Factor V Leiden. Identifying an animal
at risk for developing a thromboembolic condition may be
accomplished by any method including evaluating an animal's medical
history and standard clinical tests or assessments. In certain
embodiments, the pharmaceutical composition comprises Compound No.
957943. In certain embodiments, the pharmaceutical composition
consists or consists essentially of Compound No. 957943 and a
pharmaceutically acceptable carrier or diluent.
[0347] In certain embodiments, methods comprise administering a
pharmaceutical composition described herein to an animal with a
disease or condition of the kidney. In certain embodiments, the
condition is nephrotoxic drug exposure. In certain embodiments the
condition is a genetic or developmental malformation of the kidney,
such as that caused by a cystic kidney disease. Non-limiting
examples of cystic kidney diseases are autosomal dominant
polycystic kidney disease, autosomal recessive polycystic kidney
disease, medullary cystic kidney disease and glomerulocystic kidney
disease. In certain embodiments, the condition is primary or
secondary vesicoureteral reflux. In certain embodiments, the
condition is a urethral stricture. In certain embodiments, the
condition is a primary or secondary nephrotic syndrome. In certain
embodiments, the condition is a primary or secondary
glomerulonephritis. In certain embodiments, the condition is lupus
nephritis, giant cell arteritis, chronic urinary outflow
obstruction, or nephrolithiasis. In certain embodiments, the
condition is kidney failure. In certain embodiments, the disease is
chronic kidney disease (CKD). In certain embodiments, the disease
is end stage renal disease (ESRD). In certain embodiments, the
animal is a human subject at risk for CKD or ESRD. Subjects at risk
for CKD or ESRD include human subjects who smoke, are obese (e.g.,
body mass index greater than 30), have hypertension, have diabetes
mellitus, receive dialysis, or a combination thereof. In certain
embodiments, the human subject is a dialysis patient. The dialysis
patient may have previously received dialysis, undergoes dialysis,
will receive dialysis, or a combination thereof. In certain
embodiments, the human subject is a dialysis patient with ESRD or
CKD. Dialysis patients may be at high risk for thrombotic events.
Dialysis patients may also be at risk for bleeding events. For
example, blood vessels of ESRD patients may be compromised
resulting in an increased risk of a thrombotic event or a
hemorrhagic event. Thus, dialysis patients may benefit from
therapeutic agents that are anti-coagulatory, but do not increase
risk of bleeding, such as pharmaceutical compositions described
herein. In certain embodiments, the pharmaceutical composition
comprises Compound No. 957943. In certain embodiments, the
pharmaceutical composition consists or consists essentially of
Compound No. 957943 and a pharmaceutically acceptable carrier or
diluent.
[0348] In certain embodiments, methods comprise administering a
pharmaceutical composition described herein to an animal who has
been identified as in need of anticoagulation therapy. Examples of
such animals include, but are not limited to, those undergoing
major orthopedic surgery (e.g., hip/knee replacement or hip
fracture surgery) and patients in need of chronic treatment, such
as those suffering from arterial fibrillation to prevent stroke. In
certain embodiments, methods comprise administering an oligomeric
compound described herein, thereby prophylactically reducing a FXI
RNA or protein in an animal. Certain embodiments include treating
an animal in need thereof by administering to an animal a
therapeutically effective amount of a pharmaceutical composition
comprising a modified oligonucleotide complementary to a nucleic
acid encoding human FXI. In certain embodiments, the pharmaceutical
composition comprises Compound No. 957943. In certain embodiments,
the pharmaceutical composition consists or consists essentially of
Compound No. 957943 and a pharmaceutically acceptable carrier or
diluent.
[0349] In certain embodiments, methods comprise administering one
or more pharmaceutical compositions described herein to an animal
who has an inflammatory condition. In certain embodiments, an
inflammatory condition means any disease or condition related to an
inflammatory response to injury or stimulus characterized by
clinical signs of increased redness (rubor), temperature (calor),
swelling (tumor), pain (dolor) and/or loss of function (functio
laesa) in a tissue. In certain embodiments, examples of such
diseases, disorders, and conditions include, but are not limited
to, arthritis, colitis, diabetes, sepsis, allergic inflammation,
asthma, immunoproliferative disease, antiphospholipid syndrome,
graft-related disorder, trauma, autoimmune diseases, vasculitis, or
surgery-related disorders. Examples of arthritis include, but are
not limited to, rheumatoid arthritis, juvenile rheumatoid
arthritis, arthritis uratica, gout, chronic polyarthritis,
periarthritis humeroscapularis, cervical arthritis, lumbosacral
arthritis, osteoarthritis, psoriatic arthritis, enteropathic
arthritis and ankylosing spondylitis. Examples of colitis include,
but are not limited to, ulcerative colitis, Inflammatory Bowel
Disease (IBD) and Crohn's Disease. Examples of graft-related
disorders include, but are not limited to, graft versus host
disease (GVHD), disorders associated with graft transplantation
rejection, chronic rejection, and tissue or cell allografts or
xenografts. Examples of immunoproliferative diseases include, but
are not limited to, cancers (e.g., lung cancers) and benign
hyperplasias. Examples of autoimmune diseases include, but are not
limited to, lupus (e.g., lupus erythematosus, lupus nephritis),
Hashimoto's thyroiditis, primary myxedema, Graves' disease,
pernicious anemia, autoimmune atrophic gastritis, Addison's
disease, diabetes (e.g. insulin dependent diabetes mellitus, type I
diabetes mellitus, type II diabetes mellitus), good pasture's
syndrome, myasthenia gravis, pemphigus, Crohn's disease,
sympathetic ophthalmia, autoimmune uveitis, multiple sclerosis,
autoimmune hemolytic anemia, idiopathic thrombocytopenia, primary
biliary cirrhosis, chronic action hepatitis, ulcerative colitis,
Sjogren's syndrome, rheumatic diseases (e.g., rheumatoid
arthritis), polymyositis, scleroderma, psoriasis, and mixed
connective tissue disease. In certain embodiments, such animal has
been identified as at risk for developing an inflammatory
condition. In certain embodiments, the animal has a protein C
deficiency or a protein S deficiency. In certain embodiments, such
animals are at risk of developing an inflammatory condition due to
various genetic, situational, disease, or environmental factors. In
certain embodiments, such factors may include, but are not limited
to, familial history of inflammatory disease such as diabetes,
colitis or arthritis, exposure to allergens such as pollen,
exposure to material such as asbestos or environmental pollutants.
Identifying an animal at risk for developing an inflammatory
condition may be accomplished by any method including evaluating
the animal's medical history and standard clinical tests or
assessments. In certain embodiments, the animal has been identified
as in need of anti-inflammatory therapy. Examples of such animals
include, but are not limited to, those animals who have been
diagnosed with an inflammatory condition and those animals who have
a risk factor for developing an inflammatory condition. In certain
embodiments, the pharmaceutical composition comprises Compound No.
957943. In certain embodiments, the pharmaceutical composition
consists or consists essentially of Compound No. 957943 and a
pharmaceutically acceptable carrier or diluent.
[0350] XI. Potency and Efficacy
[0351] In certain embodiments, pharmaceutical compositions
disclosed herein are sufficiently potent to reduce FXI RNA, FXI
protein, FXI activity, or a combination thereof in an animal. In
certain embodiments, a single dosage unit of a pharmaceutical
composition disclosed herein is sufficiently potent to reduce FXI
RNA, FXI protein, FXI activity, or a combination thereof in an
animal. In certain embodiments, potency can be determined by
determining a first amount of a FXI RNA, FXI protein, or FXI
activity before administering a pharmaceutical composition
comprising an oligomeric compound disclosed herein and determining
a second amount of the FXI RNA, FXI protein, or FXI activity at a
relative timepoint after administering, and comparing the first
amount to the second amount. In certain embodiments, the relevant
timepoint after administering is 1 hour, 2 hours, 3 hours, 4 hours,
5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12
hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours,
19 hours, 20 hours, 21 hours, 22 hours, 23 hours, or 24 hours. In
certain embodiments, the relevant timepoint after administering is
1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9
days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16
days, 17 days, 18 days, 19 days, 20 days, 21 days, 22 days, 23
days, 24 days, 25 days, 26 days, 27 days, 28 days, 29 days, or 30
days. In certain embodiments, the relevant timepoint after
administering is greater than 30 days. In certain embodiments, the
relevant timepoint after administering is after a final dosage unit
of the oligomeric compound is administered to an animal. In certain
embodiments, the relevant timepoint is after an intermediate dosage
unit of the oligomeric compound is administered to the animal. An
amount of reduction in the second amount relative to the first
amount may serve as an indication of potency. The amount of
reduction may be expressed as percent reduction as demonstrated
herein.
[0352] In certain embodiments, methods comprise reducing a FXI RNA
in an animal. In certain embodiments, methods comprise reducing a
FXI protein in an animal. FXI is expressed in the liver but
secreted to the blood where it is active. Thus, FXI RNA, FXI
protein, and FXI activity levels may be measured in plasma or
serum. In certain embodiments, methods disclosed herein reduce a
FXI RNA, a FXI protein, and/or a FXI activity. FXI activity may
cause a change in blood clotting activity in the animal. The FXI
activity may cause a reduction in blood clotting activity in the
animal. Blood clotting may be measured by a standard test, for
example, but not limited to, activated partial thromboplastin time
(aPTT) test, prothrombin time (PT) test, thrombin time (TCT),
bleeding time, or presence of D-dimer in a blood sample of the
animal. In certain embodiments, a FXI RNA is reduced in a liver or
in plasma/serum of an animal by 1-100%, or a range defined by any
two of these values. In certain embodiments, a FXI RNA, FXI
protein, or FXI activity is reduced by 1%, 2%, 3%, 4%, 5%, 6%, 7%,
8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%,
22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%,
35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%,
48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%,
61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%,
74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%,
87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or
100%.
[0353] In certain embodiments, pharmaceutical compositions and
methods described herein are efficacious because they improve a
cardiovascular outcome in an animal. In certain embodiments,
pharmaceutical compositions and methods improve a cardiovascular
outcome in a population treated with a pharmaceutical composition
disclosed herein and/or according to a method disclosed herein
relative to a population that is not treated with the
pharmaceutical composition and/or according to the method. In
certain embodiments, the improved cardiovascular outcome is a
reduction of the risk of developing a thromboembolic condition. In
certain embodiments, the improved cardiovascular outcome is a
reduction in the occurrence of one or more major cardiovascular
events. Cardiovascular events include, but are not limited to,
death, myocardial infarction, reinfarction, stroke, cardiogenic
shock, pulmonary edema, cardiac arrest, and atrial dysrhythmia. In
certain embodiments, the cardiovascular event is an ischemic
stroke. In certain embodiments, the cardiovascular event is a
peripheral arterial ischemic event, e.g., amputation ischemia or
limb ischemia. In certain embodiments, the improved cardiovascular
outcome is evidenced by improved carotid intimal media thickness.
In certain embodiments, improved carotid intimal media thickness is
a decrease in thickness. In certain such embodiments, improved
carotid intimal media thickness is a prevention an increase of
intimal media thickness.
[0354] XII. Certain Comparator Compositions
[0355] In certain embodiments, Compound No: 416858, a 5-10-5 MOE
gapmer, having a sequence of (from 5' to 3') ACGGCATTGGTGCACAGTTT
(incorporated herein as SEQ ID NO: 3), wherein each internucleoside
linkage is a phosphorothioate internucleoside linkage, each
cytosine is a 5'-methyl cytosine, and each of nucleosides 1-5 and
16-20 (from 5' to 3') comprise a 2'-MOE modified sugar, which was
previously described in WO 2010/045509, incorporated herein by
reference, is a comparator compound. Compound No. 416858 was
selected as a comparator compound because it was selected for
clinical development and was confirmed to be tolerable in phase 1
clinical trials, and is currently in phase 2 clinicals. See, e.g.,
BETHUNE, et al, "Pharmacokinetics and Pharmacodynamics of
Ionis-FXI.sub.Rx, an Antisense Inhibitor of Factor XI, in Patients
with End-Stage Renal Disease on Hemodialysis," Blood
(2017)130:1116; BULLER, et al, "Factor XI antisense oligonucleotide
for prevention of venous thrombosis." N Engl J Med (2015) 372(3):
232-240 and LIU, et al, "ISIS-FXI.sub.Rx, A Novel and Specific
Antisense Inhibitor of Factor XI, Caused Significant Reduction in
FXI Antigen and Activity and Increased aPTT without Causing
Bleeding in Healthy Volunteers" Blood (2011)118:209.
[0356] In certain embodiments, Compound No. 957943 described
herein, is superior relative to Compound No. 416858 because it
demonstrates one or more improved properties, such as, potency and
tolerability. Compound No. 957943 may be dosed at lower amounts
than Compound No. 416858. Compound No. 957943 may be dosed less
frequently than Compound No. 416858. For example, as provided in
Example 5, below, in the phase 1 human clinical trial for Compound
No. 957943, individuals were administered Compound No. 957943 once
every four weeks.
[0357] For example, as provided in Example 1 (hereinbelow),
Compound No. 957943 demonstrated an ED.sub.50 for FXI RNA reduction
of 0.41 mg/kg in hFXI transgenic mice. Comparator Compound No.
416858 demonstrated an ED.sub.50 for RNA reduction of 8.35 mg/kg in
the same study. Therefore, Compound No. 957943 is demonstrably more
potent than comparator Compound No. 416858 in hFXI transgenic mice
for RNA reduction.
[0358] For example, as provided in Example 1 (hereinbelow),
Compound No. 957943 demonstrated greater FXI protein reduction at
each dosage level as compared to Compound No. 416858 when Compound
No. 957943 was dosed at one-tenth the amount of Compound No.
416858.
[0359] For example, as provided in Example 2 (hereinbelow),
Compound No. 957943 demonstrated lack of platelet reduction in
cynomolgus monkeys. Also, as provided in Example 4 (hereinbelow),
Compound No. 957943 demonstrated lack of platelet reduction in
human subjects.
[0360] For example, as provided in Example 3 (hereinbelow),
Compound No. 957943 demonstrated an EC.sub.50 for FXI RNA reduction
of 0.02 .mu.M with primer probe set RTS2966 and 0.03 .mu.M with
primer probe set RTS36807 in HepatoPac.RTM. cells. Comparator
Compound No. 416858 demonstrated an EC.sub.50 for FXI RNA reduction
of 0.98 .mu.M with primer probe set RTS2966 and 1.07 .mu.M with
primer probe set RTS36807 in the same study. Therefore, Compound
No. 957943 is demonstrably more potent than comparator Compound No.
416858 in this study.
[0361] For example, as provided in Examples 4 and 5 (provided
hereinbelow) Compound No. 957943 effectively reduced plasma
concentrations of FXI protein and FXI activity in human subjects at
all doses and time points tested when administered weekly and
monthly.
[0362] XIII. Certain Compositions
[0363] 1. Compound No. 957943
[0364] In certain embodiments, Compound No. 957943 is characterized
as an oligomeric compound consisting of a conjugate group and a
modified oligonucleotide, wherein the conjugate group is a
THA-GalNAc.sub.3 that is directly attached to the 5' end of the
modified oligonucleotide through a phosphodiester linkage,
(THA-GalNAc.sub.3)o; wherein (THA-GalNAc.sub.3)o is represented by
the following structure:
##STR00018##
and wherein the modified oligonucleotide is a 5-10-5 MOE gapmer,
having a sequence of (from 5' to 3') ACGGCATTGGTGCACAGTTT
(incorporated herein as SEQ ID NO: 3), wherein each of nucleosides
1-5 and 16-20 (from 5' to 3') comprise a 2'-MOE modification and
each of nucleosides 6-15 are 2'-deoxynucleosides, wherein the
internucleoside linkages between nucleosides 2 to 3, 3 to 4, 4 to
5, 5 to 6, 16 to 17, and 17 to 18 are phosphodiester
internucleoside linkages and the internucleoside linkages between
nucleosides 1 to 2, 6 to 7, 7 to 8, 8 to 9, 9 to 10, 10 to 11, 11
to 12, 12 to 13, 13 to 14, 14 to 15, 15 to 16, 18 to 19, and 19 to
20 are phosphorothioate internucleoside linkages, and wherein each
cytosine is a 5'-methyl cytosine.
[0365] In certain embodiments, Compound No. 957943 is characterized
by the following chemical notation: (THA-GalNAc.sub.3)o Aes mCeo
Geo Geo mCeo Ads Tds Tds Gds Gds Tds Gds mCds Ads mCds Aeo Geo Tes
Tes Te; wherein,
(THA-GalNAc.sub.3)o is represented by the following structure:
##STR00019##
and wherein,
[0366] A=an adenine nucleobase,
[0367] mC=a 5'-methyl cytosine nucleobase,
[0368] G=a guanine nucleobase,
[0369] T=a thymine nucleobase,
[0370] e=a 2'-MOE modified sugar,
[0371] d=a 2'-deoxyribose sugar,
[0372] s=a phosphorothioate internucleoside linkage, and
[0373] o=a phosphodiester internucleoside linkage.
[0374] In certain embodiments, Compound No. 957943 is represented
by the following chemical structure:
##STR00020##
[0375] In certain embodiments, the sodium salt of Compound No.
957943 is represented by the following chemical structure:
##STR00021##
EXAMPLES
[0376] The following examples illustrate certain embodiments of the
present disclosure and are not limiting. Moreover, where specific
embodiments are provided, the inventors have contemplated generic
application of those specific embodiments. For example, disclosure
of an oligonucleotide having a particular motif provides reasonable
support for additional oligonucleotides having the same or similar
motif. And, for example, where a particular high-affinity
modification appears at a particular position, other high-affinity
modifications at the same position are considered suitable, unless
otherwise indicated.
Example 1: Potency of Compound No. 416858 and Compound No. 957943
in Transgenic Mice
[0377] Oligomeric compounds No. 416858 and 957943 were tested in a
Factor XI PAC transgenic mouse model which uses bacterial P1
artificial chromosome (PAC) containing the entire WT human Factor
XI gene. The mouse model was generated from a human FXI gene
fragment containing the entire .about.24 Kb human FXI transgene as
well as 9 Kb upstream and 6 Kb downstream. The gene fragment was
microinjected into the pronucleus of fertilized mouse eggs, and the
complete BAC integration of the transgene was confirmed by PCR
using human specific primer probe sets. The established founder has
predominate liver expression of human FXI RNA and circulating human
FXI in plasma.
TABLE-US-00001 TABLE 1 Oligomeric compounds complementary to human
FXI 5' Chemistry SEQ Compound Modi- Notation ID Number fication (5'
to 3') NO: 416858 none
A.sub.es.sup.m.sub.esG.sub.esG.sub.es.sup.mC.sub.esA.sub.ds 3
T.sub.dsT.sub.dsG.sub.dsG.sub.dsT.sub.dsG.sub.ds
.sup.mC.sub.dsA.sub.ds.sup.mC.sub.dsA.sub.esG.sub.es
T.sub.esT.sub.esT.sub.e 957943 (THA-
A.sub.es.sup.mC.sub.eoG.sub.eoG.sub.eo.sup.mC.sub.eoA.sub.ds 3
GalNAc.sub.3)o T.sub.dsT.sub.dsG.sub.dsG.sub.dsT.sub.dsG.sub.ds
.sup.mC.sub.dsA.sub.ds.sup.mC.sub.dsA.sub.eoG.sub.eo
T.sub.esT.sub.esT.sub.e Subscripts: `d` represents a 2'-deoxyribose
sugar; `e` represents a 2'-MOE modified sugar. All cytosine
residues throughout each gapmer are 5'-methyl cytosines,
represented by a superscript `m'. The internucleoside linkages are
represented by subscript `o' or `s', wherein 'o' represents a
phosphodiester internucleoside linkage and `s' represents a
phosphorothioate internucleoside linkage.
Treatment
[0378] Transgenic hFXI mice were divided into groups of 8 mice
each, 4 male and 4 female. Five groups of mice were administered
1.0, 2.5, 5.0, 10.0, or 25.0 mg/kg of Compound No. 416858. Five
groups of mice were administered 0.1, 0.25, 0.50, 1.00, or 2.50
mg/kg of Compound No. 957943. Mice were administered oligomeric
compounds twice a week for two weeks by subcutaneous injection and
sacrificed at the end of the study. A group of 6 mice (3 male and 3
female) was administered PBS twice a week for two weeks and
sacrificed at the end of the study. The PBS-injected group served
as the control group to which oligomeric compound treated groups
were compared.
RNA Analysis
[0379] After two weeks, mice were sacrificed and RNA was extracted
from liver for real-time PCR analysis of measurement of RNA
expression of human Factor XI using primer probe set RTS2965
(forward sequence ACGGTGTTTGCAGACAGCAA, designated herein as SEQ ID
NO: 4; reverse sequence TGCAGATTCGGCCACAGA, designated herein as
SEQ ID NO: 5; probe sequence ACAGTGTCATGGCTCCCGATGCTTTT, designated
herein as SEQ ID NO: 6.). Results are presented as percent change
of RNA, relative to PBS control, normalized to total RNA levels
determined with Ribogreen.RTM.. Compound No. 416858 achieved an
ED.sub.50 of 8.35 mg/kg and Compound No. 957943 achieved an
ED.sub.50 of 0.41 mg/kg. ED.sub.50 values reported in Table 2 are
an average of ED.sub.50 values that were calculated based on data
from a single daily dosing.
TABLE-US-00002 TABLE 2 Percent reduction of human Factor XI RNA in
female transgenic mice Compound Daily Dose Liver Factor XI
ED.sub.50 No. (mg/kg/dose) (% control) (mg/kg) PBS -- 100 416858
1.0 80 8.35 mg/kg 2.5 85 5.0 79 10.0 40 25.0 28 957943 0.10 74 0.41
mg/kg 0.25 62 0.50 56 1.00 27 2.50 7
Protein Analysis
[0380] Reduction of FXI plasma protein was demonstrated by ELISA.
At the end of the study, blood was collected under anesthesia via
cardiac puncture into sample tubes coated with citric acid. Blood
was centrifuged at 4000 g for 15 minutes and platelet poor plasma
was collected and stored at -80.degree. C. prior to analysis.
Pooled plasma samples from all groups were analyzed by VisuLize FXI
Antigen Kit (Affinity Biologicals INC). Data are presented as the
mean of the individual samples.
TABLE-US-00003 TABLE 3 Percent reduction of human Factor XI protein
in the plasma of transgenic mice Compound Dose Plasma Factor XI No.
(mg/kg/dose) (% control) PBS -- 100 416858 1.0 81 2.5 82 5.0 71
10.0 56 25.0 30 957943 0.1 77 0.25 72 0.50 59 1.00 33 2.50 21
Example 2: Tolerability of Oligomeric Compounds Complementary to
Human Factor XI in Cynomolgus Monkeys
[0381] Compound No. 416858 was administered to cynomolgus monkeys
at 4, 8, 12, and 40 mg/kg/week by subcutaneous injection for 13
weeks, as described in Husam, et. al., "Antisense inhibition of
coagulation factor XI prolongs APTT without increased bleeding risk
in cynomolgus monkeys", Blood, 2012, 119: 2401-2408, incorporated
by reference herein in its entirety.
[0382] Compound No. 957943 was administered to groups of 14-18
cynomolgus monkeys, half male and half female, at 1, 6, and 25
mg/kg once a month or 1.5 mg/kg weekly by subcutaneous injection.
Platelet levels were measured during routine CBC measurements.
TABLE-US-00004 TABLE 4 Platelet Counts in Cynomolgus Monkeys after
Treatment with Compound No. 416858, measured at day 93 Dose
(weekly) none 4 8 12 40 (PBS) mg/kg mg/kg mg/kg mg/kg Platelets
(.times.10.sup.3/.mu.L) 506 516 452 404 357
TABLE-US-00005 TABLE 5 Platelet Counts in Cynomolgus Monkeys after
treatment with Compound No. 957943 for up to 87 days Dose 1.5 1 6
25 none mg/kg, mg/kg, mg/kg, mg/kg, (PBS) weekly monthly monthly
monthly Platelets 367 380 355 398 409 (.times.10.sup.3/.mu.L)
baseline (9 days pre-treatment) Platelets 378 366 347 372 370
(.times.10.sup.3/.mu.L) day 31 Platelets 381 386 359 391 396
(.times.10.sup.3/.mu.L) day 59 Platelets 373 369 352 378 379
(.times.10.sup.3/.mu.L) day 87
Example 3: Potency of Compound No. 416858 and Compound No. 957943
In Vitro in HepatoPac.RTM. Cells
[0383] The HepatoPac.RTM. kit is a commercially-available in vitro
liver model system available from BIOIVT that consists of
micropatterned hepatocyte "islands" co-cultured with supportive
stromal cells. A 96-well HepatoPac plate was equilibrated for 48
hours at 37.degree. C. and 10% CO.sub.2 in fresh maintenance medium
prior to treatment. Oligomeric compounds were diluted into
maintenance medium at 0.0002, 0.0020, 0.0200, 0.2000, 2.0000, or
20.0000 .mu.M for 48 hours. After 48 hours, medium was replaced
with fresh maintenance medium without additional compound. Cell
lysates were collected at 96 hours post compound addition and
analyzed by RT-PCR using primer probe set RTS2966 (forward sequence
CAGCCTGGAGCATCGTAACA, designated herein as SEQ ID NO: 7; reverse
sequence TTTATCGAGCTTCGTTATTCTGGTT designated herein as SEQ ID NO:
8; probe sequence TTGTCTACTGAAGCACACCCAAACAGGGA designated herein
as SEQ ID NO. 9.). IC.sub.50 was calculated using a linear
regression on a log/linear plot of the data in excel. Compound No.
416858 exhibited an IC.sub.50 of 0.98 .mu.M and Compound No. 957943
exhibited an IC.sub.50 of 0.02 .mu.M. Data was confirmed using a
second primer probe set RTS36807 (forward sequence
GCCAGGTAGTCTGCACTTAC, designated herein as SEQ ID NO: 10; reverse
sequence GTCCTATTCACTCTTGGCAGT, designated herein as SEQ ID NO. 11;
probe sequence CCACCCGATGGTTTACTTGTGTCCT, designated herein as SEQ
ID No. 12.). IC.sub.50 was calculated as described above. Compound
No. 416858 exhibited an IC.sub.50 of 1.07 .mu.M and Compound No.
957943 exhibited an IC.sub.50 of 0.03 .mu.M.
TABLE-US-00006 TABLE 6 Percent reduction of human Factor XI mRNA in
Hepatopac .RTM. cells Factor Factor XI (% IC.sub.50 XI (% IC.sub.50
Compound Dose control) (.mu.M) control) (.mu.M) No. (.mu.M) RTS2966
RTS2966 RTS36807 RTS36807 PBS -- 100 100 416858 0.0002 104 0.98 125
1.07 0.0020 107 110 0.0200 102 109 0.2000 93 100 2.0000 32 28
20.0000 8 6 957943 0.0002 91 0.02 101 0.03 0.0020 64 56 0.0200 62
63 0.2000 18 20 2.0000 9 7 20.0000 11 11
Example 4: Phase 1 Human Clinical Trial with Compound No.
957943--Single Dose Cohorts
[0384] Varying doses of Compound No. 957943 were evaluated in a
randomized, double-blind, placebo-controlled study to evaluate the
safety, tolerability, pharmacokinetics and pharmacodynamics of
single doses of Compound No. 957943 in healthy volunteers.
[0385] Subjects received doses according to Table 7. Four cohorts
of healthy volunteers were enrolled sequentially and randomized to
receive Compound No. 957943 or placebo by subcutaneous
injection.
TABLE-US-00007 TABLE 7 Single Dose Cohorts Treatment Healthy Total
Dose of Assignment Volunteers Doses Compound No. 957943 Placebo 8 1
0 mg A 6 1 40 mg B 6 1 60 mg C 6 1 80 mg D 6 1 120 mg
[0386] FXI protein concentrations in plasma were measured at
baseline, 8 days after treatment, 15 days after treatment, 30 days
after treatment and 60 days after treatment. Plasma FXI protein
concentrations were measured by a standard ELISA assay. To assay
FXI activity, plasma samples from test subjects were added to
plasma samples that were immunodepleted of FXI and clotting time
for each sample was assayed. Clotting time is proportional to the
level of FXI activity in the patient plasma since all other factors
are initially present at normal levels. The FXI content of the
patient plasma was determined from a reference curve prepared with
the FXI deficient plasma and varying dilutions of standard human
plasma. The reference value, which was derived from control plasma,
was 1.0 unit per milliliter.
[0387] Results of the single dose study are presented in Tables
9-10 and FIGS. 1A-1B, and FIGS. 2A-2B. Table 9 and FIGS. 1A-1B show
results of the activity assay. Table 10 and FIGS. 2A-2B show the
results of the ELISA assays.
TABLE-US-00008 TABLE 9 Plasma Factor XI Activity for Single Dose
Cohorts Baseline Day 8 Day 15 Day 30 Day 60 Placebo Mean 0.999
1.076 1.063 1.034 0.983 (N = 8) Mean SEM 0.037 0.055 0.055 0.039
0.046 % change 8.2 6.2 3.6 0.7 % change SEM 5.6 2.9 1.6 3.1 40 mg
Mean 1.07 0.718 0.605 0.593 0.702 (N = 6) Mean SEM 0.042 0.057
0.098 0.103 0.07 % change -33.1 -44.8 -45.9 -34.9 % change SEM 3.6
7.5 8.3 5.3 % change P-value <0.001 <0.001 <0.001 0.001 60
mg Mean 0.978 0.642 0.487 0.52 0.656 (N = 6) Mean SEM 0.041 0.057
0.071 0.087 0.074 % change -34.5 -50.2 -47.2 -34.8 % change SEM 4.8
6.8 8.2 6 % change P-value <0.001 <0.001 <0.001 0.003 80
mg Mean 0.987 0.588 0.446 0.417 0.647 (N = 6) Mean SEM 0.053 0.067
0.072 0.071 0.087 % change -38.8 -54.0 -57.8 -34.7 % change SEM 4.6
5.2 6.5 7.8 % change P-value 0.002 0.002 <0.001 0.001 120 mg
Mean 0.94 0.438 0.227 0.245 0.542 (N = 6) Mean SEM 0.042 0.029
0.027 0.058 0.084 % change -53.4 -75.9 -74.4 -43.1 % change SEM 2.3
2.5 5.5 7.6 % change P-value <0.001 <0.001 <0.001 0.001
Input datasets: ADSL and ADEFF. Note: Baseline is defined as the
last non-missing measurement prior to the first dose of study drug.
.sup.[A] ANOVA test P-value, .sup.[T] Wilcoxon rank sum t
approximation test two-sided P-value, .sup.[E] Wilcoxon rank sum
exact test two-sided P-value.
TABLE-US-00009 TABLE 10 Plasma Factor XI Protein Concentrations for
Single Dose Cohorts Baseline Day 8 Day 15 Day 30 Day 60 Placebo
Mean 1.19 1.22 1.22 1.22 1.18 (N = 8) Mean SEM 0.07 0.06 0.08 0.1
0.04 % change 3.3 3.1 3.9 3.0 % change SEM 4.6 5.8 8.5 6.2 40 mg
Mean 1.14 0.68 0.54 0.52 0.81 (N = 6) Mean SEM 0.03 0.09 0.1 0.09
0.1 % change -41.2 -53.8 -55.1 -29.7 % change SEM 6.8 7.6 7.6 8.6 %
change P-value <0.001 <0.001 <0.001 0.014 60 mg Mean 0.93
0.61 0.45 0.44 0.68 (N = 6) Mean SEM 0.04 0.07 0.08 0.1 0.08 %
change -35.0 -52.1 -53.5 -26.9 % change SEM 5.3 7.4 9.5 6.8 %
change P-value <0.001 <0.001 0.003 0.018 80 mg Mean 1.14 0.56
0.36 0.4 0.69 (N = 6) Mean SEM 0.08 0.08 0.06 0.08 0.1 % change
-49.2 -67.0 -64.9 -38.9 % change SEM 5.4 4.6 6.9 9.2 % change
P-value 0.002 0.002 <0.001 0.005 120 mg Mean 1.1 0.44 0.22 0.2
0.54 (N = 6) Mean SEM 0.05 0.02 0.03 0.05 0.08 % change -59.8 -80.0
-82.4 -51.5 % change SEM 2.2 2.1 4.3 6.9 % change P-value <0.001
<0.001 <0.001 0.001 Input datasets: ADSL and ADEFF. Note:
Baseline is defined as the last non-missing measurement prior to
the first dose of study drug. [A] ANOVA test P-value, [T] Wilcoxon
rank sum t approximation test two-sided P-value, [E] Wilcoxon rank
sum exact test two-sided P-value.
[0388] Notably, all doses reduced plasma FXI protein concentration
and activity by an average of at least 30% at 8 days after
receiving the single dose, including the lowest dose of 40 mg.
Also, it is noted that plasma FXI protein concentration and
activity was reduced by an average of at least 50% at 8 days after
receiving the single dose of 120 mg. Plasma FXI protein
concentration and activity were further reduced by an average of at
least 30% of baseline levels by 15 days after receiving the 120 mg
dose. This reduction was maintained at 30 days after dosing. These
data suggest that a monthly dosing regimen is sufficient to obtain
therapeutic effects of a single dose of Compound No. 957943.
Safety and Tolerability Evaluations
[0389] Patient safety was monitored closely during the study.
Safety and tolerability evaluations included: physical examination,
vital signs (HR, BP, orthostatic changes, weight), ECG, adverse
events and concomitant medications, plasma laboratory tests
(clinical chemistry, hematology), urinalysis, and complete blood
counts (CBC). Platelet levels were measured during routine CBC
measurements. Overall, Compound No. 975943 was well-tolerated.
There were no safety concerns in vital signs including heart rate
and blood pressure, and no clinically relevant changes in liver
chemistry, renal function, or platelet values. There were no
deaths, no serious adverse events or spontaneous bleeding
events.
Example 5: Phase 1 Human Clinical Trial with Compound No. 957943
Multiple Dose Cohorts
[0390] Varying doses of Compound No. 957943 were evaluated in a
randomized, double-blind, placebo-controlled study to evaluate the
safety, tolerability, pharmacokinetics and pharmacodynamics of
multiple doses of Compound No. 957943 in healthy volunteers.
Subjects received doses according to Table 11. Healthy volunteers
enrolled in the weekly multiple-dose treatment cohorts (AA, BB, and
CC) received six subcutaneous doses of Study Drug (Compound No.
957943 or placebo) starting on Week 1, Day 1 followed by
once-weekly subcutaneous administration during Weeks 2 to 6 (Days
8, 15, 22, 29, and 36). Healthy volunteers enrolled in the
multiple-dose treatment cohort (DDD) received four subcutaneous
doses of Study Drug starting on Week 1, Day 1 followed every four
weeks with SC administration during Week 5 (Day 29), Week 9 (Day
57), and Week 13 (Day 85).
TABLE-US-00010 TABLE 11 Multiple Dose Cohorts Cohort and Healthy
Total Dose of Dose Level Volunteers Doses Compound No. 957943 AA:10
mg weekly 8 6 60 mg for 6 weeks (3:1) BB:20 mg weekly 8 6 120 mg
for 6 weeks (3:1) CC:30 mg weekly 8 6 180 mg for 6 weeks (3:1)
DDD:80 mg every 10 4 320 mg four weeks for 13 weeks (3:2)
[0391] Plasma FXI concentrations for cohorts AA, BB and CC were
measured by a standard ELISA assay at various time points. Results
of the ELISA assay are presented in Table 12, FIG. 3A and FIG. 3B.
Plasma FXI activity was also measured (as described in Example 4)
at various time points. Results of the activity assay are presented
in Table 13, FIG. 4A and FIG. 4B.
TABLE-US-00011 TABLE 12 Plasma Factor XI Activity for Multiple Dose
Cohorts AA, BB, CC Baseline Day 15 Day 29 Day 43 Day 64 Day 78 Day
92 Placebo Mean 0.962 0.96 0.91 0.91 0.89 0.856 0.906 (N = 6) Mean
SEM 0.074 0.078 0.06 0.07 0.041 0.07 0.073 % change -0.2 -4.6 -5.1
-6.0 -11.4 -5.8 % change SEM 2.9 3.5 2.2 4.1 5 7.3 Mean 1.035 0.89
0.635 0.515 0.615 0.637 0.75 10 mg Mean SEM 0.089 0.099 0.095 0.113
0.119 0.101 0.092 weekly % change -13.8 -38.2 -51.3 -41.6 -38.1
-26.8 (N = 6) % change SEM 7.8 9.6 8.6 9.1 10 8.9 % change P-value
0.093 0.065 0.002 0.009 0.082 0.126 20 mg Mean 0.98 0.775 0.608
0.498 0.494 0.568 0.822 weekly Mean SEM 0.083 0.091 0.14 0.136
0.082 0.096 0.091 (N = 6) % change -20.6 -40.8 -51.4 -50.8 -43.7
-17.3 % change SEM 7.3 10.2 11.6 7.7 6.8 8.2 % change P-value 0.065
0.017 0.004 0.004 0.008 0.31 30 mg Mean 0.812 0.448 0.22 0.15 0.215
0.372 0.49 weekly Mean SEM 0.054 0.036 0.028 0.021 0.03 0.053 0.069
(N = 6) % change -44.6 -73.2 -81.7 -74.1 -55.1 -41.1 % change SEM
2.1 2.6 2.2 2.2 4.7 5.6 % change P-value 0.004 0.004 0.002 0.002
0.004 0.004 Input datasets: ADSL and ADEFF. Note: Baseline is
defined as the last non-missing measurement prior to the first dose
of study drug. [A] ANOVA test P-value, [T] Wilcoxon rank sum t
approximation test two-sided P-value, [E] Wilcoxon rank sum exact
test two-sided P-value.
TABLE-US-00012 TABLE 13 Plasma Factor XI Protein Concentrations for
Multiple Dose Cohorts AA, BB, CC Baseline Day 15 Day 29 Day 43 Day
64 Day 78 Day 92 Placebo Mean 1.13 1.08 1.08 1.08 1.08 1.01 1.03 (N
= 6) Mean SEM 0.06 0.08 0.08 0.08 0.08 0.05 0.06 % change -4.6 -5.1
-4.9 -4.6 -9.1 -7.3 % change SEM 3.5 3.6 3.4 4.6 3 1.6 10 mg Mean
1.17 0.94 0.74 0.54 0.68 0.78 0.9 weekly Mean SEM 0.12 0.12 0.13
0.11 0.14 0.14 0.12 (N = 6) % change -20.1 -38.5 -55.8 -44.1 -35.0
-23.6 % change SEM 5.3 5.9 5.8 7.1 7.1 4.2 % change P-value 0.041
0.002 0.002 0.004 0.017 0.017 20 mg Mean 1.24 0.84 0.64 0.56 0.58
0.72 0.89 weekly Mean SEM 0.16 0.14 0.2 0.15 0.11 0.11 0.14 (N = 6)
% change -31.5 -53.0 -57.1 -55.1 -43.9 -31.6 % change SEM 8.3 10.1
10.6 7.3 6.2 4.6 % change P-value 0.065 0.004 0.004 0.004 0.008
0.008 30 mg Mean 1.05 0.51 0.23 0.15 0.23 0.39 0.57 weekly Mean SEM
0.08 0.04 0.04 0.03 0.04 0.06 0.07 (N = 6) % change -51.0 -78.7
-85.5 -78.2 -63.0 -46.3 % change SEM 2.3 2.8 2.3 2.9 4.1 5.3 %
change P-value 0.004 0.004 0.002 0.002 0.004 0.004 Input datasets:
ADSL and ADEFF. Note: Baseline is defined as the last non-missing
measurement prior to the first dose of study drug. [A] ANOVA test
P-value, [T] Wilcoxon rank sum t approximation test two-sided
P-value, [E] Wilcoxon rank sum exact test two-sided P-value.
[0392] Plasma FXI concentrations for cohort DDD were measured by a
standard ELISA assay at various time points. Results of the ELISA
assay are presented in Table 14 and FIG. 5A and FIG. 5B. Plasma FXI
activity was measured with an assay conducted in FXI-depleted
plasma as described in Example 4 at various time points. FXI
activity is presented in Table 15, FIG. 6A and FIG. 6B.
TABLE-US-00013 TABLE 14 Plasma Factor XI Activity for Multiple Dose
Cohort DDD Baseline Day 15 Day 29 Day 43 Day 71 Day 99 Day 113
Placebo Mean 0.975 0.958 0.95 0.953 1.03 0.898 0.838 (N = 4) Mean
SEM 0.095 0.153 0.114 0.115 0.118 0.115 0.085 % change -3.0 -3.0
-2.9 -2.1 -8.5 -14.1 % change SEM 7.3 3.2 3.5 3.5 4.4 2 80 mg Mean
1.145 0.467 0.422 0.238 0.192 0.147 0.198 every Mean SEM 0.073
0.099 0.085 0.048 0.051 0.028 0.039 4 weeks % change -60.8 -63.7
-79.6 -83.9 -87.3 -82.8 (N = 6) % change SEM 6.4 7.2 3.9 3.9 2.4
3.4 % change P-value 0.01 0.01 0.01 0.024 0.01 0.01 Input datasets:
ADSL and ADEFF. Note: Baseline is defined as the last non-missing
measurement prior to the first dose of study drug. [A] ANOVA test
P-value, [T] Wilcoxon rank sum t approximation test two-sided
P-value, [E] Wilcoxon rank sum exact test two-sided P-value.
TABLE-US-00014 TABLE 15 Plasma Factor XI Protein Concentrations for
Multiple Dose Cohort DDD Baseline Day 15 Day 29 Day 43 Day 71 Day
99 Day 113 Placebo Mean 1.09 1.17 1.02 1.06 1.15 1.1 1.01 (N = 4)
Mean SEM 0.18 0.12 0.14 0.14 0.16 0.16 0.12 % change 12.0 -3.9 -0.7
-5.6 3.2 -4.4 % change SEM 11.3 7.7 3.8 3.1 6.6 6 80 mg Mean 1.33
0.51 0.45 0.26 0.23 0.19 0.25 every Mean SEM 0.15 0.13 0.1 0.06
0.08 0.05 0.05 4 weeks % change -63.8 -66.1 -81.1 -83.9 -85.5 -80.6
(N = 6) % change SEM 6.9 7 3.7 4.5 3.1 3.9 % change P-value 0.01
0.01 0.01 0.024 0.01 0.01 Input datasets: ADSL and ADEFF. Note:
Baseline is defined as the last non-missing measurement prior to
the first dose of study drug. [A] ANOVA test P-value, [T] Wilcoxon
rank sum t approximation test two-sided P-value, [E] Wilcoxon rank
sum exact test two-sided P-value.
[0393] FXI plasma protein and activity was reduced in all subjects
receiving Compound No. 957943 at all doses at all time points.
Results of these studies support a monthly dosing regimen.
Safety and Tolerability Evaluations
[0394] Patient safety was monitored closely during the study.
Safety and tolerability evaluations included: physical examination,
vital signs (HR, BP, orthostatic changes, weight), ECG, adverse
events and concomitant medications, plasma laboratory tests
(clinical chemistry, hematology), urinalysis, and complete blood
counts (CBC). Platelet levels were measured during routine CBC
measurements. Overall, Compound No. 975943 was well-tolerated.
There were no safety concerns in vital signs including heart rate
and blood pressure, and no clinically relevant changes in liver
chemistry, renal function, or platelet values. No deaths,
spontaneous bleeding, or serious adverse events were observed.
Example 6: Phase 2 Human Clinical Trial with Compound No.
957943
[0395] Multiple doses of Compound No. 957943 are evaluated in a
Phase 2 multicenter, randomized, placebo-controlled study in ESRD
patients receiving hemodialysis. Patients are randomized to receive
subcutaneous treatment (low, mid, high dose) with either Compound
No. 957943 or placebo. All standard of care hemodialysis therapies
as prescribed by their providers are continued, except
anticoagulants or antiplatelet medications other than
acetylsalicylic acid (e.g., aspirin) up to 150 mg daily.
[0396] All cohorts consist of a screening and approximately 6
months treatment period followed by a post-treatment follow-up
period.
[0397] Pharmacodynamics and efficacy are assessed at multiple time
points. Patient safety is monitored closely during the study.
Safety and tolerability evaluations may include e.g. physical
examination, vital signs, adverse events, concomitant medications,
and plasma laboratory tests.
Sequence CWU 1
1
12126001DNAHomo sapien 1atgccctcca taggctttca gaccttcttc aaggccaaag
ggaaggcctt catggaaata 60taattatgtg aaacacccca gaattttttc acaaactttc
ctgcctataa acgccaggtc 120ctaccactgt ccagtgtccc acttcccact
gtccagtggg aagactccct ccaggacaaa 180accacccttt ccttcaggga
tgtgacgtgc tgtgctttcg ttgacagtca tgcagctgct 240agacatgtca
cttcctagct ctccattcgg gtctgtgtcc caggacctaa agaaagctaa
300aagaagccgg gcgtggtggc tcatgcctgt catcccagca ttttgggagg
ctgaggctgg 360cggatcactg gagatcagga gtttgagacc agtctgacca
gcatggtgaa accctgtctc 420tactaaaaat acaaaaaaag aaaagaaaaa
agaaaaaaaa ataagctggg cgtggtggcg 480cgggcctgta gtcccagcta
ctccggaggc ggaggcagga caatcacttg aacctggaag 540gcggaggttg
cagtgagctg agatcgcgtc attgcactcc agcctgtgcg acgagagact
600ctgtctcaaa agaaagaaag aaagagagaa agaaagaaag aaagaaaaga
aagaaagaaa 660gaagaaagaa agaaaagaaa gaaagtaaga agcaagcaag
ctgaaatagc ttatttttct 720gttttaaaaa atagactttt agtgttcaca
agtaagaata aaaaaaaaaa aagcttggcc 780ttaggaggaa tcctatctgc
ttaggcctct gagaggcagc gtcacctgaa gggaaactga 840ctgggcaaga
gccaggctct taggagctgt ctgtactttg cttcccatcc gtccgctctc
900ccccagcagc caaagtctcc tccctccatt atattgcaaa tcaatgaatc
cattagactt 960tccatgtttt ctttttagac tttgagattc aaggtcagct
agaattaatg gttaacagct 1020gacaggtgaa agtgtggtta aatgcagctt
tggctagcag gcacacaggc aaaatcaagt 1080tctacatctg tccctgtgta
tgtcacttgt ttgaatacga aataaaatta aaaaaataaa 1140ttcagtgtat
tgagaaagca agcaattctc tcaaggtata tttctgacat actaagattt
1200taacgacttt cacaaatatg ctgtactgag agagaatgtt acataacatt
gagaactagt 1260acaagtaaat attaaagtga agtgaccatt tcctacacaa
gctcattcag aggaggatga 1320agaccatttt ggaggaagaa aagcaccctt
attaagaatt gcagcaagta agccaacaag 1380gtcttttcag gtacagtttc
agaacttact atttaacatt cctctcaagc aaatacgcct 1440tgaaatgctt
tttttaaatc ataggaattt aaaaacactt tacaatagag aatgattgat
1500ttttaaaatg tgtctgattt agctttgtag agatgttccg ctaatatcca
taactaatct 1560gagaggaaat gtggaacaac agaagagtaa cagtgtctac
tcagtaacaa gcgttttacg 1620agttaaaaag acatccaaat gcagtactga
aaaatcagaa gtcttgattt gtctcactga 1680tgactccgtt tttcctagag
cagtctgttt aatgcttact ggagataaat agatttatag 1740gtgaccaaga
caatcgatta atgtatcagc cacagacttt tttaaataga aaattttcta
1800agtaggaaat cattcatagc tctttgaaag atatagggag aggcctcaag
gaaagaaaga 1860aaggaaaaaa attgggaaag gaaacaaaga tgaaaaattg
gggtggggag agcggtcaga 1920tggtggccat gagaaggatc tgaacacaga
gagcggcggg gccggcgggg aaggagggag 1980gaggggagag cgctgcttcc
ctgtgggttc cggcttctgc agagctgtaa gagttgaatg 2040ccacacacag
tcacactaag gaatgctcca ggattgggaa agaaaattca acattataat
2100gagaacactg tgaatgctat tgaattaact actcccctct ctccctattt
cttgtaagtc 2160ttagtgtcag taaactaatt ataaatttac attttatgtt
ctaaaagcat gcaccttttt 2220ctcattgtag gatgattttc ttatatcaag
tggtacattt cattttattt acttcagttt 2280ctggtggtaa gtagagtgtt
atcttaacta tgggctggga gagggaaatc acactgcaat 2340ctccacacat
gtgggagaat cccacaccat ttatgccggg aaggaaataa aatgttttta
2400ttaacttcct gcctgaggct ccagaggttt tcaaagcagg gtaggaattg
aggtgaaaaa 2460attgtttgta ctggtaggaa tctgtgtcta tatgtgtacc
atatctacat ccatgcctac 2520atacatcatt catttaaatg atatttaaaa
gcaatattaa aaagaagaaa tcttgatttg 2580cctcactaat tagttggttt
ttcataaagc agtcagttca atggaacata cacacacaca 2640taaaatagga
attttgtact gaaaaatgtt atgtaactat tcatacacag tttatgcatt
2700tcaataattt aactaccctg ttaattctcc accctatgtc acatctgcta
aatttgttta 2760taaattactt agccaatatt aagtaggata tatcagtaca
ggtattgttt agttccatga 2820ccattcttta tttaaaatga aagctgaagc
attcctttag aaaaatagct tatgtctaca 2880tgttatattt aatttctgat
actattgcta gcattttaag catagtaaat tctttttgct 2940ggcctatgtg
aaaaaaggca aacctaggtg aagatattaa aagtaaattc cacttaatat
3000attaaattac acagctcata atttcaattt tctctgtgac aagactactg
ctcaggttat 3060aatattcctt tgttataatt atctggaaat tgtattgtta
tttcattgat aaatatgtca 3120gcgctaaata atttgatata ttttgaacac
tggacccaat aactacataa acaattgacc 3180ctgaatcagg ggttccacat
ccatggattt aaacaacatt gaagtgaaat tttttaaaaa 3240aaattatatc
tgcactcaac atgtgcagag ttttttcttg tcattattcc cgaaacaaca
3300cagtataaca acaatttgca tagtatttac attgtactag gtattataag
taatctagag 3360atgatttaaa gtacacagga ggctgtgagt aggttatatg
taaatactat gccattttct 3420atcagggact tgagcgttca tggattttgg
tatctgcagg aggtctggca gccaatccct 3480catagatatc aaggaatgat
agtgtacata tttaggtcct ttatacatgc acatggaaca 3540tttacaaata
ttgaccacac acaagcctct gaaacaaggc tcaacaaatt ttaaggggtt
3600aaaattatac tgatcatgtt attcaatcac cgtgaatggg agctagaata
ataataaaaa 3660gggaattgaa aatctgcaaa cgtttgaaaa ttgagtattt
ataatcaccg tgaatgggag 3720ctagaataat aataaaaagg gaattgaaaa
tctgcaaatg tttgaaaatt gagtatttat 3780acctttttag attaagatga
atcagagatg aaatcacaat ggtaattaga aaaggaattg 3840tacaataatg
aaagtacagt atatcaaaac tttgagatgt aaccgaatca gtgttgagat
3900caaatgtata gatttacatg tatacattag aatacagaaa agggaaaaat
taatgatata 3960agatgacaag aagatagaga atagcaactt aagtgctaag
aaattagaag gcagaaagaa 4020gaaatcgcag aaattaatta aatggaagac
acacatattt gagaggtcaa cataacccct 4080ttctagccca ttttataagg
tcagggtaat cttgacacca aaatctgata ggaaaattct 4140gaggaaaaaa
agtcacaggt cttttatctt atgaacatat gtacaaatgt cctaaacaaa
4200atattatcca acttaaccca gtagattgtt aaaaaaatta aaggatgatt
acattgtgat 4260attgtgacaa agttgggttt attccaggaa tgaaaagttg
atttaacatt caaaaatcaa 4320tagatgtgtg acttcggggt acataccgaa
aaaaaagaag ggaaagcagg aactcacaca 4380gatatacttg tacacccatc
tcatagcagc attattcaca ctaacccaaa ggtggaagca 4440gcccatgtgt
ttatcaacag gtgactggat aaagtgtggt acatacatac aaaggaatat
4500tactcaaccc taaaaagaaa ggaaattctg acacatgcta tgacatggat
gaaccttgag 4560gacattacgc taagtgaaat aagccagtca caaaagaaca
aatactgtgg gatctcactg 4620ataggaggta cttagggtag tcaatttcat
caagacggaa attagaacag aggttgccag 4680gggccgaggg gccaggggaa
gggctgaggg ttggtgttta atgggcacag agcatcagct 4740ggcgaagttg
aaaagttctg gaggtggatg gtgctgatgg ttgcacagca atgtaaatac
4800ccttagtgcc acagaactgt acactcaaga tggctacatg gcacatggta
tgctatctgt 4860cataataaaa aataaaaata attttaaatg ttaatatgtg
ccgaaaaatg cttgctaaaa 4920ttctattaat gatcaaaact ttaaataaac
tggaaatgga agagaacttt catcagctat 4980taaacgatat tttaaacaaa
agcaaacaaa aatccaaacc aaaaatcttc aagaaacata 5040atacgtaata
gcggaatatt gaaagctttc ccaagggatt gagaatagga caagaatact
5100tcccattatt tccatttact cttgactgga ggtactcgtt ggtgcggtaa
ggaagggaaa 5160taaaaggaaa acgattagga ggaacataag ctctcgtttt
tcacagatga tgtgattgag 5220tacctagaaa aatccaaaat taaccatcac
ataaattatt tgagccaatt aattaaatga 5280gtaagaagtc tcgatacaac
gtcaatatac aacagcaagc acttgctcgt tcgcaaaagc 5340agatgaacta
gtttttactg acactagaag cagcatgtgt tccatgtgtt ccacatccat
5400gcactcaaac aaccttgacg tgaaatattt tttaaaaaat tgtatctgta
ctcaacgtgt 5460gcagagtttt tttccaaaat tgttttccaa aaatttttaa
atgtttatat gtttaggggg 5520tgtaagtgca gatttcttac atacatatat
tatgcggcgg tgaagtgccg gcttttggtg 5580tacccatcac ccaagtagtg
aacacagcct ccaacaggta atttttcaac gctcacccca 5640ctcccatcct
cccatctagt ggcatattga aaatcaattt gtcttgtaaa attaaataat
5700ccaattagga gggggatata ttctaaggaa attagtgcat gatgcacaca
cacacacaca 5760cacagaacac gtgtgtgcgc atgtgcacat gagagagagt
gagagagaaa ctgggtcttg 5820ctctgtcgcc caggctggat tgcagtggta
aaatcacagc tcactgcagc ctcaaactcc 5880caggctcagg agatcctcct
acctcagcct cccgagtagc tgggattaca ggtggaaaca 5940accatgccca
gctagtattt tttttttttt ttgtattttt tatagagaca gggtcttgcc
6000atgttgccca ggctggtctt gaactcctca gctcaagcaa tctacctacc
ttagcctccc 6060aaagtgctgg gattacacgc atgagccact gcgcccactc
cgcattatta aatatagaac 6120atttatttga ttcatcagtt aatattcttc
ttaaaagtac tattttaatg tagcaagatt 6180gctttccacc aaaaggtggg
gtttccacgg tggggtttcc aaattattct caatggggtg 6240aggatgtgtg
ttatcacacc cccgagccat cagatgctgt cagaaagtga tcactctgaa
6300gtctttgttt caaataagca cagggtttgg ataaagagac gcaattagga
aaggaaaaag 6360cagaaggctc gttccagacc tggatgagat cctaaaaagc
agcagctttt gccagtaaag 6420atccttgaaa tgattcaatt accctcaaag
cactccttgt ctccaagaca atcactcata 6480agcacaattc cattgaagcc
aacgtaccat tttgtgattt tcgtttccac ctgaggctgt 6540tcattcaata
aactcacata aaagtgttta ttgccttgat ttccaaattc aggcgtattt
6600cctggtaagt agagctactt gccttgcctt tatgagatta ccacctaact
agatgtatgc 6660ccagtaaaat ccaacataac gcatgccatg tactacatca
cagaatgtgt gactcagttg 6720ttgaaggaca cctgctttga aggaggggac
attactacgg tcttcacacc aagcgccaag 6780tactgccagg tagtctgcac
ttaccaccca agatgtttac tcttcacttt cacggcggaa 6840tcaccatctg
aggatcccac ccgatggtaa atgcttatgt ttctacatcg aggagacaga
6900tttttaaagg gagattgcta ttcttaacac atttccatct aacattttat
aaaatttaac 6960attaacaact ggaagataaa ttgtctttca gttgaaatat
tgttacagaa agaagtgatg 7020gtgtttacgc aatttagaaa agaataaata
tgcctccaag tgtagacttt ccagcctctc 7080ctatagtctc atattaatgg
tatgtttctt ctgtttgtct caatttttac acttcttaaa 7140catttcacac
tttgcttttc tatcgattat taatttttgt cgtgcttctc aacaaactgg
7200aacttctccc aactatttta ttagaaaaaa ataaaatatt taaaaagaaa
atttgaaaaa 7260aagtacagta cagtgatccc cctgccacca ccaaaaccgc
aatgtttgct atatttgtac 7320agcaatatac aatatacaat acctatattt
gtatacatgt ttaaccgttt gaaataattt 7380taaaacatga cactttaccc
ctaaatactt cagcatgcac tatcctacac aaaagacata 7440cgaaatttaa
caagaattcc tttatattat ttcagttttc cctcaaatat aatttatagt
7500aattaaccag aatcttacca agagtcactc actgcattgg gtggtttgtc
aggtttaaaa 7560cattttaaac agtccaccaa ccatttgtat tcccatcctg
agcttgaaaa tttaataata 7620agcctttctg tagaattaag ttttcaacat
ctttattatt gctacattca caggcattta 7680tgtagcaccc agaacttata
aaatttacta ttccagaacc tagagcaggg attggcaaat 7740gtcttcttaa
taacgcagag taaatatgtt aggctttgtg ggcaaaaccc acagtaaagc
7800caaggatatt atttaagtat ttatgtcacc acttaaaatg taacaatttg
aaaatataaa 7860aatcattttg tatagctaac aggctaaaca gaaacacaca
gatttttggt tgcattttac 7920caacaggtcc tagttgacac attcctgttt
gttcctatta gaaaggagta ttacatgcag 7980tctcttaagt gtagggatat
tgaagtaaaa aacaaactca gaatcttgct aagaaaatat 8040ttgttttggc
atgagataaa gtagtttgtt tccttctttt tggctttctg tgtgctgact
8100tttaagatcc attattttaa aaacataaat tcctattcat taatatgtat
tttttaaaaa 8160aacaggttta cttgtgtcct gaaagacagt gttacagaaa
cactgccaag agtgaatagg 8220acagcagcga tttctgggta ttctttcaag
caatgctcac accaaataag cggtaagata 8280tgttctcaga atcaacaaat
accagctgtg atgtacacat atcgccacat cggatgtggt 8340tttaaggcta
tgaaatgaaa cactgctatg tggaaataaa cccccttaat gaagttcttt
8400cagtgtagag tataaactag tatacataca tgcctgccct ccaacacact
gtaaaaacct 8460ctttacctca tagaaagaca tatcttacta cctcacttcc
catcatttat ttatattctt 8520tctatttccc agcctaaaat cttaaatgaa
agtctttttt tttttgagac agggtctcac 8580tctgttgtcc aggctggagt
gcagtggtgc aatcacagct cactccagcc tcaacctcct 8640gggttcaagt
gatcttcctg ccttagcctc tggagtagct gggaccagag gcatgcacca
8700acattcccag ctaatttgtt catttttcct agagacaggg tctcactgta
ttgcccaggc 8760tggtctcaaa ctcttggcct caagtgatct gcccgcctcg
gccttccaga gtgccgggat 8820tccatggtgc ccagtcgaaa ttctttatta
aacgtattaa tccaaattga aaggagcaaa 8880tataaaggtt gaagtgacac
ttgtcttaat agtgaataga tacttgaatc agttaattag 8940cgaaataatc
agtgcagtta gggagaagag aggctaggtc agaaaatcaa aatgtgaatt
9000tacaagtcta aaattgttac agtgtaagaa ggacattggc attcttttac
tgcttccatt 9060caagaataag aattttgcag attaatataa cgaaagacct
ctgaggaaag gtgggtgaaa 9120aagttgaaag gatgagtcag gagggacagt
tgcttaggtc attgccccta gaatctggaa 9180ggtactcatg tcttctgctt
ttatttccag cttgcaacaa agacatttat gtggacctag 9240acatgaaggg
cataaactat aacagctcag ttgccaagag tgctcaagaa tgccaagaaa
9300gatgcacgga tgacgtccac tgccactttt tcacgtacgc cacaaggcag
tttcccagcc 9360tggagcatcg gtgagtgagt cccaggacat tcgagtggtc
gatgaaaaac agaatcgtga 9420tttactaaaa agcttttgcc atcaacttta
tgccagaatt tattttgaac ccctaaaaga 9480catttctata atagtactcc
tagttttctt catgaaaaat acacttaaag cctaatttgg 9540atgcatttca
tttatggtaa ggagtctatc ttttaataac actgtcagaa aaatatatat
9600acttggctaa tttcaaaagc gctacacttt taaattggca cttttgaaac
agctgcaatt 9660ggtatgattg tcagtgccct tcccagtcta aaaaatgtta
cagtctaaca gaataaaaat 9720aaaaacctac tctctctctc tctaaataac
agttccttac ctaagacaaa atactcatgt 9780aaaaagtctt atcctgctcc
atactggatt ttgaaatatt tcaaggataa atctatcaca 9840taaggattta
aaaattatct gatctctaat aaccaaatct gtgttctcat ctttaaaaat
9900ttactaggga aatagattat taatttgtat attcagaaat atttgagatg
atttagattt 9960tcatagtaaa ctgcatttat ctggaatcaa cagaaaagtg
aaaaacattc aaattactaa 10020tacttgcgtt ttaacattgg attttaacat
tctgctctcc acattcacaa agaggagtga 10080acagaaagca aacaaagcat
caacgagtta tttcaaaaac aacagtggtg aaaaacacac 10140acaccaaacc
cctaaattca tgatttgact tgtaaggctt atctttagct cagctcagac
10200gacagctttt atgtctaaga cttaacagaa tgtgaactgc aagacaagaa
attggaggtt 10260tctaagcaag ataaagttaa gtcattaaaa gtaagaagga
cttagccagg cgcggtggct 10320cacacctgta atcccagcac tttgggaggc
cgaggcgggc agatcacctg aggtcaggag 10380ttcgacacca gcctgaccaa
tatggtgaaa ccccgtctct actaaaaaag aatacaaaaa 10440ttagccaggc
gcggtggtgg gcgcttgtaa tcccagctac ttgggaggct gagacaagag
10500aatcgcttga acccaggagg cggaggttgc agtgagccaa gatcgtgcca
ttgcactcca 10560acctgggcaa cagagtgaga ctccgtctca aataaaaaaa
aacaaaaaat gagaagggct 10620tgagaagtca ttcattcatg cactctcctt
cttcatgtgg tcactctctc aagctgtcat 10680tatactgaag aagaaataaa
cttacacaat tcacaggtgc ttagcaacac tgctgggacc 10740atgcccagcc
attcagcctc ccagatggat gcttcggggt ctcgcaggtc ctctctccaa
10800aggggacttt cttaatatct catgtttttt cctccttgca gttggaagaa
taagacactt 10860ttcctttttc tttttattca gtaacatttg tctactgaag
cacacccaaa cagggacacc 10920aaccagaata acgaagctcg ataaagtggt
gtctggattt tcactgaaat cctgtgcact 10980ttctaatctg ggtaattatc
gacttcttga tgatgtaatt caaccattaa atatgctgat 11040gattacagta
gatctcactc aggataccag cttatgctca cgatgaaacg gacccaaaga
11100tctttacctt cttcatgtga tagatttcat catgtcctat acagttagat
cctctattta 11160aatttccagt ttaaaataat catgccattt tcttctaaat
aaaaaaaaat taaaagatct 11220tgggatacac ttaaattttt taatatggaa
tttacacata ctgtgaccgg aattttcctg 11280atagctggtg aattgagtcc
ctgacatagt tcttccgtcg cgcagcttgt attagggaca 11340ttttccctaa
tacggtgttt gcagacagca acatcgacag tgtcatggct cccgatgctt
11400ttgtctgtgg ccgaatctgc actcatcatc ccggttgctt gttttttacc
ttcttttccc 11460aggaatggcc caaagaatct caaaggtaag gagttaacaa
gtaaggataa tttgttatct 11520tctaaaaata gctgatcaaa atccatcatt
aaaaattcca agtaactaaa aatttactct 11580aaatgtcagt ataggataaa
agttgcaaag aatttctagc ccctctccct ttctattccc 11640cacctactta
ccacaaaccc aacattaccg aggactcttt tttttttttt tttttttttg
11700agatggagtc tcgctctgct gcccaggctg gagtgcggtg gcatgatttc
agctcactgc 11760aaccttcgcc tcccaggtcc aagcgattct cctgcctcag
cctcctgagt agctgggact 11820acaggcatgg gacaccacgc ccagtaattt
tttttgtatt tttagtagag atggggtttc 11880accatgttgg ccaggctggt
ctcaaactcc tgacttcagg tgatccacct gcctcggcct 11940cccaaagtgc
tgagattaca gggttgggcc accgtgcccg gccagtaaat tttaaaataa
12000atataaatat tacttcacct aaataaattt taggtacagg tacagttgtg
ttacatggat 12060atactgtgta gtggtgaagt ctgggctttc agtgtagcca
aatagtatac attattccca 12120ttagataatt tctcctgcct caccctcctc
tatcctccca actctctgag tctccaatag 12180ctttcattcc actgtctctg
tgcctgtgta cactttattt agctcccact taaaagtgag 12240aatctacaat
atttgacttt ctgtttctga gttgtttcac ttgagataat agcctccagt
12300tctatccatg ttgctgcaaa agatatgatt ttatgatttt tttatggcta
agtaacattc 12360tatagtatat tctatacacc acattttctt tatccattca
tttgctgata gactcttagg 12420ttgatccata tctttgttat tgtgaatagt
gctgcagtaa acatgtgagt gtaggtatct 12480ttgtgacatg attttttttt
ctttcttttc ctttggctat gtacccagta gtgggattgc 12540tggaggtctc
caccttggca aaggggcagt tgtctagttc taaatgaaat gaagagattc
12600catttccatt tcatgacaac taaaagacaa ttcagtccaa tgctttgttt
aaaataattg 12660aaccaggaac agaaagagct gaaaatgtca gtgaaatgtc
aaacccaaag tggagagaga 12720gagagaggtg gaaatgaatg tctccatcaa
gtgggttagg gtggtggtgg agggaggttt 12780gagaattgag tccctgtttc
cttaccaatg gaaattaaca taggacatca gaaacagcat 12840ggaaaatttc
tttgattatc aaaagtacac tagctaaggt tgctgtccct cttttattta
12900tttatttatt tgagacaggg tcttgctctg tcactcagat ttggttgcac
tgggtgtgat 12960ctcagcccac tgcagcctcc acttcctagg ctcaagcaat
ccacccgtct catcctccca 13020agttgctggg accacaggtg tgcaccacca
cccccagcta atgtttgttt ttttgtagag 13080acagagtttc gccatgttgg
ccaggctggt ctagaactcc tggcctcaag caatccacct 13140gccttggtct
cccacagtgc agggattaca agcctgagcc actatgccca gcttgctatt
13200cctcattgac aacattcact taaacaagca aacaaatatc cgtaaaatta
agtcagcttt 13260aaaacctggc ttgtatatat ctctgaattg gaactctcag
agccctcagc acttgcctga 13320ttggcctcct gttgatgaaa agttgtcctc
cagacaactc agccaaggga ggccctgtgt 13380gccttgccta tcacatgagc
ctcactttcc actgagtgag gctgtcattt cagaagcacc 13440gggtctgtca
catgaaaata tatctgttac catcacttac taaacaaatt tagtagaatt
13500tgtttggtgc ttattatgta tcaggcattg ttctgaaggc tggggatacc
atttagtgaa 13560ctaaatcgac aaaaactttg cctattggac cagagtggag
atgagagaga agtgaccaga 13620tcgatctgtg ttatcagcag acagccaata
agatttgctg ataggttgga ccacggattg 13680tgaaggagtc agtgatagca
ccaagacttt tggcccaagg aactggaaag atggaaatgt 13740caatgacaga
atgtggagga tttcgcaagc agcagagtgt aggagaagca atgcccttgg
13800ctcatctagg gcaaagggtt gaattggcaa ctggaaatag aagtcaaatt
cagaagagag 13860gtttatgtgg acatctgtgt ttaggtgtca ccaatataca
gctgatactt tttaaaaatc 13920tagtgaaatt atcaaagggt taaatgcaga
gagggaagaa agtaggtcca aagatcaagc 13980cttgggacat tggaagttta
gaaataagaa agatgtcatt gtcacttgta attttgtgct 14040agtcactgct
ctttttcttt gtcttattac cttactgacc aattcctaga ataggaataa
14100cacatttgat ctttaataca gtatgtgata ggaacatggc ttctataagc
ccaaacttgg 14160caatttaaat ttaaatttat taaaattaaa taaaactggc
tgggtgcagt ggctcacgcc 14220tgtaatccca gcactttggg aggccgaggc
ggtgaatcac gaggtcagga gtacgagacc 14280atcctggcca acatggtgaa
accccgtctc tactaaaaat acaaaagtta gccaggtgtg 14340gtggcatgtg
cctgtaatcc cagctactca agaggctgag gcaggagaat cacttgaacc
14400cgggaggcgg aggttgcagt gagccaagat cgcgacactg cactccagcc
tgggtgacaa 14460gagaaagact atgtctcaaa aaaaaaaaaa attttaaata
aaactaaaaa ttcaccttcc 14520cagctgtgtt agccacattt caagtgccca
atagccacat gtagttggca gttactgtat 14580tggatggcac aggcatagaa
tatttccatc actgcagaaa gatctatgga cagtcttgct 14640ctagactgtg
attttgtcta cttaggaaaa gtcactcttt tccaggaaga tgcttccagg
14700gtgtggagta aacgacggac ttcacccatc tccttataaa cctattgcaa
ccctggatag 14760gaggtttcta ggtagcatga agactcttga atctttaaga
taggctggga gtgttgaaag 14820gaaggaaatg aaaagggaag atactaggaa
gactgacaat agagcaagct cagaaaattt 14880ttatggaggt gatttagtta
gaaaatttgt ctgcaaccta agggccatgg agtgtgactc 14940catggtttat
gggtgtaact ccgtggttta tgaagagtac tttcaaaata ggaaaatctt
15000cacaactaag tgctagcatg agctgacttt actttctcta
ggtgctgtaa aaatgttttt 15060atgtgtttga tatgatatat ttctacttcc
cttttgtttt tgttagaaat ctttgtctcc 15120ttaaaacatc tgagagtgga
ttgcccagta cacgcattaa aaagagcaaa gctctttctg 15180gtttcagtct
acaaagctgc aggcacagca tcccaggtaa actgagagtt ctgcattctg
15240gctgagagtg accagccccg aggaggctga tacatgctga gggagggtct
cactctgaca 15300tgtggtctgc tgtctagtgt tctgccattc ttcattttac
catgacactg atttcttggg 15360agaagaactg gatattgttg ctgcaaaaag
tcacgaggcc tgccagaaac tgtgcaccaa 15420tgccgtccgc tgccagtttt
ttacctatac cccagcccaa gcatcctgca acgaagggaa 15480gtaagccata
tgaagggtta tgcagacacc cttgtcccgt ctgcctgtga ggtgcattat
15540gtttataccg ttttgtttcc aactgcaggg gcaagtgtta cttaaagctt
tcttcaaacg 15600gatctccaac taaaatactt cacgggagag gaggcatctc
tggatacaca ttaaggttgt 15660gtaaaatgga taatggtgag tataatgtca
cttgaaaaaa tatagctgaa ggaattattc 15720catgcttcat acatcacaat
caagactgtc agttatagcc acagaaggga gaacattcag 15780gaaataacaa
attttgcaat tttctattat tttcactcct gtcactcaag ctgaccatgt
15840tttaaaggta aatattgagg cttgactaaa ctgtacattg cctagtatta
actaaatata 15900tgctttaatt tgacacattt atacacctgg catttctgtt
ttctttcttt cttttttttt 15960tttttttttt tgagacacag tctcactctg
ttgcccaggc tggagtgcag tgatgtggtc 16020agtgttcact gcagcctctg
gcttgaactc ctggactcga acaatcctcc caccttagct 16080ccctgagtag
ctggggctac aggcgtgcac cacaacactc ggctaatttg ttgtagagat
16140gggatcttgc catgttgccc aggctggtct tgaactcctg ggctcaagca
atcctcccgc 16200tttggcctcc caaagttctg ggattacagg cttcagccac
tgcacccagc cacaactggc 16260atttcctaat gagacccaga ctctgccaac
actcctgtta tcaaggccag taatctttcc 16320tattatcctt tgtaagggaa
ttatcgatca gcactttggt tagtcaaatt actaattttc 16380ttccaaaaat
tgtgtatcta ttcaagaaat actggagcac ctccttttta gggtctttat
16440tcagattcca tgcagggttc tggagactta gggattggca aaggttaagg
taaaacttta 16500ctagtaacaa tgagtgtggt aggatggaaa taagtgttta
gattacacga gacctgtaat 16560aacataaaag ttacaataaa aatccaacca
gcagtgtttc catcccagtg gccaatttct 16620gagcatagtc acgagagatg
ctttgtaggc aaacagaatt gttctaaggg acaaaatccc 16680caacaggaaa
gaacacacca caaacactcc tgcttaaatt accatagtaa cttaggatgg
16740ctttactata tattgattta cataaagctg catgttctta tactgtttat
tgccaaatgt 16800ccaatattac aatacactat aaggtgcaac tgagatttaa
tgaataaaat ggaagtaaca 16860atcctgcctc gtgatagttt tagaagcaca
aaaacattct gtgtcagatt atctgctgta 16920ccgagaaggc gaatcaatcc
ttaatttctg agaacttgtt ttgtagaaca taaagacgtt 16980atattgcctc
caacactggt atcctaacta acagactatg ccttcctaga gcttagaggc
17040gctccgatga aaatctctgg atggctcaag acttcttaaa aagcaagtca
attacgtcgt 17100atctcataca ttctgttttc ctcacaataa atttccctaa
gacaagaagg agcattcggc 17160accattctgt tgtctttctt ctacttctaa
gcgttagaag ggacacttag caaatgttgc 17220tgttaagtaa tgttgacatg
gtttaataaa atgggaatga gcacgtatac ctcaatacat 17280tgcagactgc
attttccccc ttccttcttc attatggttt tctctgttgg atttatagac
17340tctggcctgt agaagttaca gaatatgcca ggtatagatt gatagctaca
ggagaaaatg 17400taagataaag gaaaataaag tcttacgtct tttcagtgca
actttggagg gagtgaatta 17460gataactagg tttttactgc gctgtatttg
atgaaataac cccctaatgt gaaagggaat 17520agctgcgtga gatatttatg
gtgccttgtc tgtcactggt ctacaatgta acttaacttt 17580ctgaagatag
atagcagcac cattaaaata aacatttctt accacaaaat atgattctaa
17640acacatattt tcagcatttc gtttaaactg agaaacagca taggatgaac
ctcaaggcct 17700ctcacctgga ccttgagtta tttctaaaat atcttagtta
ctatttacct attaattttc 17760ctaaaattta cctttatgac gcttcccacc
ttgcagaaat tccagaatag atggccctcc 17820aaaatgaatg ttcacccttc
cggctctaaa atgagagcct ctgttcaggc ttccgaagtc 17880acatcgtgct
cgttctcacc tcagttgctg ttagctgctg tttccttccg aactccttct
17940ctcatcctct cctcctattt gaaatctgcc ccagaattat acactcattt
tcctaccaag 18000gaaaaaaagg cctagaaagg ttgttttaca cccacaaaac
tagtgaatgg accttctagg 18060acccggcttc tcatcagtga ttcttctgtt
aacttagact cctcccttag ctcaggacgc 18120ggagccttct gagcacctga
gcctggttat tctaaatgtg atctgggcac agcacattga 18180catcacctgg
gagcttgtta gaaagaattt caggacccac acagatgtta ctgagtcaga
18240atctgcattt tgaaagctac acaggcaacc cacaggcaca taaattttga
gtcgcatagg 18300tgtgtgcgtg tgtgtgcgca tgtatgtgtg cgtgtgtttg
tgtgcacgtg tgtgtgtgtg 18360tgtctagaat actgctgtct acgaacaccc
tattcccatc catctgtgtt ccatggctcc 18420aaccgggagg gtgggttctt
gtgtcgggca tccagtaagt agaaatagag ggcactgtcc 18480tgtctaggca
gccccaagag aaaagaaaca gagggatgag cctgagtcaa agtccctgaa
18540aagtaccaag gaccccagag aatcccaaac tgtcaacaag gccaaaggtc
agaggaagtt 18600catagcagat cagtcttact ttggacgtgg gtggagcagg
agtgactggg atcatggtca 18660gcagagtcac tgggacaggg caggaactgg
acaatggctg cagcctgcgg gcaaggtgct 18720tgccttttct ttctaagagc
agttctcaaa cgccagcagg gctggttcaa acacagatgg 18780ctgagcttcc
aggctggagt ttctcattca ctggatctga ggttgggctg aagaatgtgc
18840atttctaaca cgttcccagg tgacgctgtt ggtctggaga ctgcacttga
caaccactgg 18900tttaaaaaca ccattcacgt tatcatttga aggagggtaa
gacagccttg tagtaccaca 18960caaggagggc tacattctta ggggtgtgta
attacaagat gacttagtca attccatttt 19020tcatgtgcat gttttgcttt
ggcagcttga ttataaagtc tctgtaactc agggtcatga 19080taaactattg
acttgaggaa aggttttctt cttgttcctg aaggagcata attactgatg
19140gaaaggaaga tgtaggaagc tgctcatcac aatgcttctg ttgcagagtg
taccaccaaa 19200atcaagccca ggatcgttgg aggaactgcg tctgttcgtg
gtgagtggcc gtggcaggtg 19260accctgcaca caacctcacc cactcagaga
cacctgtgtg gaggctccat cattggaaac 19320cagtggatat taacagccgc
tcactgtttc tatgggtcag taccacggct gtttttatta 19380gttcatcttc
ttcacacatt tataaaaaat attactagca tgttaggaaa taaatacttt
19440aaccaattag attgtcttat ttgcaaaatt aattaattgc ttcagtggta
aaaaacgcaa 19500aaaggaagag ctcatggtct cccagcatca gaacaggtgc
aggtacaagg ctgcttgact 19560gcctgctatt ccgcttccca tttaaccgca
ttcacatccc caagggcctt catgttattc 19620cctgcaagag catacctccc
tctgtgcctc gctctgtgca ctgtgcccgg aactaaactc 19680acagaggatt
taccattgtc tgaatcaaat ttctaatatg tgtgtgtgtg tgtgtgcgtg
19740tgtgtttaaa tacagaaagt ggccaggtgt ggtggttcac gcctgtcatc
tcagcacttt 19800gggtggctga ggtgggagga ttgctcaatg ccaggagtct
gaggtcagcc tgggcaacat 19860agtgagacct cgtctctaaa aaatatttaa
aaactagcct ggcatggtgt tttgtgtgcc 19920tgtagcacca gctgctcaga
agtctgaggt aggaggattg cttgagccca agagttcaag 19980ggtgcagtga
gttactacag tgccactgtg ctccagcctg agcaacagag caagacccta
20040tctctaagta aataaataaa atacagaacg agttcggtat gcatcctcac
attggattcc 20100ttacttaggt cactttcagc gttggccaaa caaaaaggct
ccagctgggg gtatatatat 20160tccagggaag ttaagttggc caaactttcc
gtttgctact tcagtatcct ccgagttgtt 20220tccagacaca gttttgtgtg
ctttttagtt ttggttttct tttttaatca atctgcagca 20280cactcatgtt
aaactcaaga acacatgtga ggccaagagt tcccgttacc tgtcacatat
20340gggaatggag tagcagaaag cctagtttct gtcacagcta gttactggcg
agattgagac 20400tgagtccagc ggtcttcgtg tgtgtgtgtg cgtgtgtgtc
tgtgcagtgt gcgtgtgtgt 20460gcgtgtctgt gtgtgcgtgt gcgggtgtgt
gcgtgtgtgt gtgttggagc acggagggag 20520tgctcattca ttttttgtgt
ataatggatt ttctttatag ggtgaatatg ttttttatcc 20580cgaaaaatct
taggataaaa tcactttttt ctacctaaat gtccatcatt ggcagaaaat
20640attagtaata attaaacagc cacacacttc acaatgtctg ggaattattt
ttagtaaagg 20700aaatttcttt ccctctgttg tttgctcctt agggtagagt
cacctaagat tttgcgtgtc 20760tacagtggca ttttaaatca atctgaaata
aaagaggaca catctttctt tggggttcaa 20820gaaataataa tccatgatca
gtataaaatg gcagaaagcg ggtatgatat tgccttgttg 20880aaactggaaa
ccacagtgaa ttacacaggt acggagaatt ttatccggaa agttgtctcc
20940aatggtgaac tggataaaat gtttaacact actagactta cggcctgacc
ctgccaatct 21000ctccatgcgt tatcatcatg aaagggagag ggcctggaat
gctagtcatt cactctgcta 21060aggctgacac actttcctgg ctattgaaac
ttattttggg aatgtgggta aagagatacg 21120ttttcctgag tcttcttcag
gtgcatagaa tgacataatt tcataatact ttggaatagt 21180aaagataatt
tagtctaaag ataatttatt aaagataatt tagggatgaa ggattgaagg
21240ttagaacaat taagcaactt gtgcaggatc aaagtgagtt ggatgaggag
ttagcggtga 21300gggtgaggct tgtctctctc tcgccctctc atcctggcac
atgtgcgata tcgtgctgaa 21360cctgagggag gaaaatacac gacaacaagg
caaaaaatga atatagtaaa caaagaaaac 21420acagataatg tacagtggaa
gaagagtctc ttctggaaaa gaggatatat tttgcgtctc 21480atatttaaac
cacgattttt taaatttaga ttctcaacga cccatatgcc tgccttccaa
21540aggagataga aatgtaatat acactgattg ctgggtgact ggatgggggt
acagaaaact 21600aagaggtaaa aatgatgttg ttatatgtgc tccatcctag
aaatgaagag cggaaccttt 21660tctgccctgt caagtcatgt agctgaagca
caactcgagt cacactactc agttgcagga 21720agcggattaa taaagatgga
gaggcaaaaa tcacccaagt gaggctggtg cctcatatgt 21780ttgattggaa
attttaaatg tgactaaatc tctttaaaga ctaattatat ttaatgaagt
21840ttaatgtgaa gcctagcact tttcagtaaa tgttctagcc tgctatccaa
ttactttctt 21900gggaagtcat tccagttaga gtcataatta atttttgaac
ttaattaaca ttaacaaaat 21960ggtacacgca atagtgggaa taatgtcttc
ttcatacttg taattataaa aggtctgtga 22020agtaaatcta acattttttc
cttctagatt tttatataga catgagtttt gtgttgttgt 22080tgttttgaga
tggactctcg ctctgtcgcc caggctggag tgcagtggca cgatctcggc
22140tcactgttac ctccacctcc cgggttcaag tgattctcct gcctcagcct
cccgagtagc 22200tgggattata ggtacccaac caccacccca agctaatttt
ttgtattttt agtagagacg 22260gggtttcatc atgttagcca ggatggtctc
aatctcctga cctcgtgatc cacctgcctc 22320ggccttccaa agtgctggaa
ttacaggcgt gagcccccac acccgtccat gatttttatt 22380ttaaatatat
gtggcccagc accactggtg gctcacgcct gtaatcccag cactttggga
22440ggccaagatg ggtggatcac ttgaggtcag gagttcaaga ctggcctggc
caacatggtg 22500aaaccctgtc tctactaaaa atacaaaaat tagctgggca
tggtggtgtg tgcctgtaat 22560cccagctact cgggaggctg aggcaagata
atcgcttgaa cttgggaggt ggaggtagca 22620gtgagctgag attgcaccac
tgcactccag cctgggcgac agaaagagac tccgtctcaa 22680ttaaaaatat
atatatatat atatttatat gtatgcatat atgtttatgt gtattgtgta
22740tggttattct acaaacgaac caaaaaaatt tttttcagac aaaatacaaa
atactctcca 22800gaaagccaag atacccttag tgaccaacga agagtgccag
aagagataca gaggacataa 22860aataacccat aagatgatct gtgccggcta
cagggaagga gggaaggacg cttgcaaggt 22920aacagagtgt tcttagccaa
tggaatatat gcaaattgga atgcttaatg cgttggggtt 22980tttttgtttg
ttttgttttt tttgtttgtt tttttttgag acagagtctc gctctgttgc
23040ccaggctgga gtgcagtggc tcgatctcag ctcactgcaa gctctgcctc
ccaggttcac 23100gccattctcc tgcctcagcc tcccaaatag ctgggactac
aggcgccagc taccaagccc 23160agctagcgtc tttttttttt ttagttttag
tagagacggg gtttcaccat gttggccagg 23220atggtctcga tctcctgacc
tcatgatctg cctgcctggg cctcccaaag tgctgggatt 23280acaggcgtga
gccaccgcgc cgggccgctt aatgcatttt aaaaagcagt cttctgccaa
23340tgagcaggga acacagtgta tttgtttgac ttagactgaa atcaaaagca
aggagattga 23400ctggatgaac gcaagcaccc aggttctctg cagtatatta
aggggccaag acaacatttt 23460aggcaaaatc agcctgagca agatgtgctg
aagatgggaa gcgtctgagt tgatctgtgc 23520accttttctt gtctcccctc
gttctaggga gattcgggag gccctctgtc ctgcaaacac 23580aatgaggtct
ggcatctggt aggcatcacg agctggggcg aaggctgtgc tcaaagggag
23640cggccaggtg tttacaccaa cgtggtcgag tacgtggact ggattctgga
gaaaactcaa 23700gcagtgtgaa tgggttccca ggggccattg gagtccctga
aggacccagg atttgctggg 23760agagggtgtt gagttcactg tgccagcatg
cttcctccac agtaacacgc tgaaggggct 23820tggtgtttgt aagaaaatgc
tagaagaaaa caaactgtca caagttgtta tgtccaaaac 23880tcccgttcta
tgatcgttgt agtttgtttg agcattcagt ctctttgttt ttgatcacgc
23940ttctatggag tccaagaatt accataaggc aatatttctg aagattacta
tataggcaga 24000tatagcagaa aataaccaag tagtggcagt ggggatcagg
cagaagaact ggtaaaagaa 24060gccaccataa atagatttgt tcgatgaaag
atgaaaactg gaagaaagga gaacaaagac 24120agtcttcacc attttgcagg
aatctacact ctgcctatgt gaacacattt cttttgtaaa 24180gaaagaaatt
gattgcattt aatggcagat tttcagaata gtcaggaatt cttgtcattt
24240ccattttaaa atatatatta aaaaaaatca gttcgagtag acacgagcta
agagtgaatg 24300tgaagataac agaatttctg tgtggaagag gattacaagc
agcaatttac ctggaagtga 24360taccttaggg gcaatcttga agatacactt
tcctgaaaaa tgatttgtga tggattgtat 24420atttatttaa aatatcttgg
gaggggaggc tgatggagat agggagcatg ctcaaacctc 24480cctaagacaa
gctgctgctg tgactatggg ctcccaaaga gctagatcgt atatttattt
24540gacaaaaatc accatagact gcatccatac tacagagaaa aaacaattag
ggcgcaaatg 24600gatagttaca gtaaagtctt cagcaagcag ctgcctgtat
tctaagcact gggattttct 24660gtttcgtgca aatatttatc tcattattgt
tgtgatctag ttcaataacc tagaatttga 24720attgtcacca catagctttc
aatctgtgcc aacaactata caattcatca agtgtgattt 24780tttttttttt
tttttgagat gaagtctcac cctgttgccc aagctggagt gcagtggtgt
24840gatctcggct cactgtaaac tctacctcct ggattcaagc gattgtcctg
cctcagtctc 24900ccaagtagct gagattacag gcacatgcca ccatgcccgg
ctaatttttg tatttttagt 24960agagacgggg tttcactatg ttggccaggc
tggtcttgaa ctcctgacct cgtgatctgc 25020ccacctcggc ctctcaaagt
gctgggatta caggtgtgag tcactgcgtc tggccatgga 25080aaatatttat
tgagcacaat tatgtgagag catcatgctg agctttgaag atacagtggt
25140gagcaaacat atatcctggc ttcatgaaga ttatactcta gttaacatga
gcaacaaaat 25200aaaataatca cacaaaatat ataggttcaa gctgaaatga
gtggctgcac cagattctat 25260gagataagaa aggaagaagg acatttttca
ccaagttcaa agactgggat acaaaggaat 25320ttgtcctgac aaaggcaaaa
caaaaacaac aacaaacaaa aaacccaaaa gagcaaaatg 25380acagtagaac
ataacggggc cagatcaaaa atgctgacag gttcccaaaa gaataaaatg
25440acggtaggac atgacggggc cagatccaaa atgctgacag gttcaaacaa
aattggaatt 25500gaaaatcaga gtgcgttcaa gagtatcaaa caatactatc
ttgttacttg cttattacct 25560tagtagactg gaagcaacac ttcacacaaa
aaaagggttt ggatgtaatt tcggataaga 25620agagatgttt ctgtaaagtc
tttcctgaga agcatattat ttgagaaaaa cacatatttc 25680tgtttttagt
atttcacttt gtataatgtc ttaatttttg aagagctggt atattcctat
25740gattcattaa tgaaagttct ataagatata aaatatacaa tgaggagatc
tcctcttctg 25800taccagaaga gtgcacattc tacacactgc gtagcacctt
tctcacttac gttctgtctg 25860ggcacacttc tgattgacac gcagagggct
ctctctgtct ggggatattt ctgatgggta 25920ccgagagagc ttcctctatc
ttgggttatt tctgatgcgt agagaagggc tgcctctgtc 25980cattatggaa
ggctggtgtt c 2600123278DNAHomo sapien 2aggcacacag gcaaaatcaa
gttctacatc tgtccctgtg tatgtcactt gtttgaatac 60gaaataaaat taaaaaaata
aattcagtgt attgagaaag caagcaattc tctcaaggta 120tatttctgac
atactaagat tttaacgact ttcacaaata tgctgtactg agagagaatg
180ttacataaca ttgagaacta gtacaagtaa atattaaagt gaagtgacca
tttcctacac 240aagctcattc agaggaggat gaagaccatt ttggaggaag
aaaagcaccc ttattaagaa 300ttgcagcaag taagccaaca aggtcttttc
aggatgattt tcttatatca agtggtacat 360ttcattttat ttacttcagt
ttctggtgaa tgtgtgactc agttgttgaa ggacacctgc 420tttgaaggag
gggacattac tacggtcttc acaccaagcg ccaagtactg ccaggtagtc
480tgcacttacc acccaagatg tttactcttc actttcacgg cggaatcacc
atctgaggat 540cccacccgat ggtttacttg tgtcctgaaa gacagtgtta
cagaaacact gccaagagtg 600aataggacag cagcgatttc tgggtattct
ttcaagcaat gctcacacca aataagcgct 660tgcaacaaag acatttatgt
ggacctagac atgaagggca taaactataa cagctcagtt 720gccaagagtg
ctcaagaatg ccaagaaaga tgcacggatg acgtccactg ccactttttc
780acgtacgcca caaggcagtt tcccagcctg gagcatcgta acatttgtct
actgaagcac 840acccaaacag ggacaccaac cagaataacg aagctcgata
aagtggtgtc tggattttca 900ctgaaatcct gtgcactttc taatctggct
tgtattaggg acattttccc taatacggtg 960tttgcagaca gcaacatcga
cagtgtcatg gctcccgatg cttttgtctg tggccgaatc 1020tgcactcatc
atcccggttg cttgtttttt accttctttt cccaggaatg gcccaaagaa
1080tctcaaagaa atctttgtct ccttaaaaca tctgagagtg gattgcccag
tacacgcatt 1140aaaaagagca aagctctttc tggtttcagt ctacaaagct
gcaggcacag catcccagtg 1200ttctgccatt cttcatttta ccatgacact
gatttcttgg gagaagaact ggatattgtt 1260gctgcaaaaa gtcacgaggc
ctgccagaaa ctgtgcacca atgccgtccg ctgccagttt 1320tttacctata
ccccagccca agcatcctgc aacgaaggga agggcaagtg ttacttaaag
1380ctttcttcaa acggatctcc aactaaaata cttcacggga gaggaggcat
ctctggatac 1440acattaaggt tgtgtaaaat ggataatgag tgtaccacca
aaatcaagcc caggatcgtt 1500ggaggaactg cgtctgttcg tggtgagtgg
ccgtggcagg tgaccctgca cacaacctca 1560cccactcaga gacacctgtg
tggaggctcc atcattggaa accagtggat attaacagcc 1620gctcactgtt
tctatggggt agagtcacct aagattttgc gtgtctacag tggcatttta
1680aatcaatctg aaataaaaga ggacacatct ttctttgggg ttcaagaaat
aataatccat 1740gatcagtata aaatggcaga aagcgggtat gatattgcct
tgttgaaact ggaaaccaca 1800gtgaattaca cagattctca acgacccata
tgcctgcctt ccaaaggaga tagaaatgta 1860atatacactg attgctgggt
gactggatgg gggtacagaa aactaagaga caaaatacaa 1920aatactctcc
agaaagccaa gataccctta gtgaccaacg aagagtgcca gaagagatac
1980agaggacata aaataaccca taagatgatc tgtgccggct acagggaagg
agggaaggac 2040gcttgcaagg gagattcggg aggccctctg tcctgcaaac
acaatgaggt ctggcatctg 2100gtaggcatca cgagctgggg cgaaggctgt
gctcaaaggg agcggccagg tgtttacacc 2160aacgtggtcg agtacgtgga
ctggattctg gagaaaactc aagcagtgtg aatgggttcc 2220caggggccat
tggagtccct gaaggaccca ggatttgctg ggagagggtg ttgagttcac
2280tgtgccagca tgcttcctcc acagtaacac gctgaagggg cttggtgttt
gtaagaaaat 2340gctagaagaa aacaaactgt cacaagttgt tatgtccaaa
actcccgttc tatgatcgtt 2400gtagtttgtt tgagcattca gtctctttgt
ttttgatcac gcttctatgg agtccaagaa 2460ttaccataag gcaatatttc
tgaagattac tatataggca gatatagcag aaaataacca 2520agtagtggca
gtggggatca ggcagaagaa ctggtaaaag aagccaccat aaatagattt
2580gttcgatgaa agatgaaaac tggaagaaag gagaacaaag acagtcttca
ccattttgca 2640ggaatctaca ctctgcctat gtgaacacat ttcttttgta
aagaaagaaa ttgattgcat 2700ttaatggcag attttcagaa tagtcaggaa
ttcttgtcat ttccatttta aaatatatat 2760taaaaaaaat cagttcgagt
agacacgagc taagagtgaa tgtgaagata acagaatttc 2820tgtgtggaag
aggattacaa gcagcaattt acctggaagt gataccttag gggcaatctt
2880gaagatacac tttcctgaaa aatgatttgt gatggattgt atatttattt
aaaatatctt 2940gggaggggag gctgatggag atagggagca tgctcaaacc
tccctaagac aagctgctgc 3000tgtgactatg ggctcccaaa gagctagatc
gtatatttat ttgacaaaaa tcaccataga 3060ctgcatccat actacagaga
aaaaacaatt agggcgcaaa tggatagtta cagtaaagtc 3120ttcagcaagc
agctgcctgt attctaagca ctgggatttt ctgtttcgtg caaatattta
3180tctcattatt gttgtgatct agttcaataa cctagaattt gaattgtcac
cacatagctt 3240tcaatctgtg ccaacaacta tacaattcat caagtgtg
3278320DNAArtificial sequenceSynthetic oligonucleotide 3acggcattgg
tgcacagttt 20420DNAArtificial sequencePrimer 4acggtgtttg cagacagcaa
20518DNAArtificial sequencePrimer 5tgcagattcg gccacaga
18626DNAArtificial sequenceProbe 6acagtgtcat ggctcccgat gctttt
26720DNAArtificial sequencePrimer 7cagcctggag catcgtaaca
20825DNAArtificial sequencePrimer 8tttatcgagc ttcgttattc tggtt
25929DNAArtificial sequenceProbe 9ttgtctactg aagcacaccc aaacaggga
291020DNAArtificial sequencePrimer 10gccaggtagt ctgcacttac
201121DNAArtificial sequencePrimer 11gtcctattca ctcttggcag t
211225DNAArtificial sequenceProbe 12ccacccgatg gtttacttgt gtcct
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