U.S. patent application number 16/053540 was filed with the patent office on 2019-02-14 for anti-pd-l1 combinations for treating tumors.
The applicant listed for this patent is BIRDIE BIOPHARMACEUTICALS, INC.. Invention is credited to Lixin Li.
Application Number | 20190048084 16/053540 |
Document ID | / |
Family ID | 55063593 |
Filed Date | 2019-02-14 |
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United States Patent
Application |
20190048084 |
Kind Code |
A1 |
Li; Lixin |
February 14, 2019 |
ANTI-PD-L1 COMBINATIONS FOR TREATING TUMORS
Abstract
The present invention relates to therapeutic combinations and
methods for treating cancers using combination therapy.
Inventors: |
Li; Lixin; (Beijing,
CN) |
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Applicant: |
Name |
City |
State |
Country |
Type |
BIRDIE BIOPHARMACEUTICALS, INC. |
Grand Cayman |
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KY |
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Family ID: |
55063593 |
Appl. No.: |
16/053540 |
Filed: |
August 2, 2018 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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15401843 |
Jan 9, 2017 |
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16053540 |
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PCT/CN2015/083585 |
Jul 8, 2015 |
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15401843 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 31/7064 20130101;
A61K 31/4745 20130101; A61K 31/55 20130101; A61K 31/4375 20130101;
C07K 16/3069 20130101; C07K 16/2827 20130101; C07K 16/3046
20130101; C07K 16/303 20130101; A61K 31/519 20130101; C07K 2317/76
20130101; C07K 16/3038 20130101; C07K 16/3023 20130101; A61K
39/39558 20130101; C07K 16/3061 20130101; A61K 31/4985 20130101;
C07K 16/3015 20130101; A61K 31/5513 20130101; A61K 31/522 20130101;
A61K 31/52 20130101; A61P 35/00 20180101; C07K 16/3053 20130101;
A61K 2039/505 20130101; A61K 45/06 20130101; A61K 31/708 20130101;
C07K 16/2818 20130101; A61K 39/39558 20130101; A61K 2300/00
20130101; A61K 31/4745 20130101; A61K 2300/00 20130101; A61K 31/708
20130101; A61K 2300/00 20130101; A61K 31/4375 20130101; A61K
2300/00 20130101; A61K 31/55 20130101; A61K 2300/00 20130101; A61K
31/5513 20130101; A61K 2300/00 20130101; A61K 31/522 20130101; A61K
2300/00 20130101; A61K 31/4985 20130101; A61K 2300/00 20130101;
A61K 31/52 20130101; A61K 2300/00 20130101; A61K 31/7064 20130101;
A61K 2300/00 20130101; A61K 31/519 20130101; A61K 2300/00
20130101 |
International
Class: |
C07K 16/28 20060101
C07K016/28; C07K 16/30 20060101 C07K016/30; A61K 39/395 20060101
A61K039/395; A61K 45/06 20060101 A61K045/06; A61K 31/4985 20060101
A61K031/4985; A61K 31/519 20060101 A61K031/519; A61K 31/52 20060101
A61K031/52; A61K 31/522 20060101 A61K031/522; A61K 31/55 20060101
A61K031/55; A61K 31/5513 20060101 A61K031/5513; A61K 31/7064
20060101 A61K031/7064 |
Foreign Application Data
Date |
Code |
Application Number |
Jul 9, 2014 |
CN |
201410325480.9 |
Sep 1, 2014 |
CN |
201410440824.0 |
Claims
1. A combination, comprising: (i) an effective amount of a
PD-L/PD-1 Axis antagonist antibody; and (ii) an effective amount of
an immunotherapeutic, wherein the immunotherapeutic is
4-amino-2-(ethoxymethyl)-a,a-di-methyl-1H-imidazo[4,5-c]quinoline-1-ethan-
ol; wherein the PD-L/PD-1 Axis antagonist antibody and the
immunotherapeutic are not covalently linked to each other.
2. The combination of claim 1, wherein said PD-L/PD-1 Axis
antagonist is selected from the group consisting of a PD-1 binding
antagonist, a PD-L1 binding antagonist and a PD-L2 binding
antagonist.
3. The combination of claim 2, wherein the PD-L/PD-1 Axis
antagonist is a PD-1 binding antagonist.
4. The combination of claim 3, wherein the PD-1 binding antagonist
is an antibody fragment.
5. The combination of claim 5, wherein the PD-1 binding antagonist
is MDX-1106, Merck 3475, CT-011, AMP-224, or AMP-514.
6. The combination of claim 2, wherein the PD-L/PD-1 Axis
antagonist is a PD-L1 binding antagonist.
7. The combination of claim 6, wherein the PD-L1 binding antagonist
is an antibody.
8. The combination of claim 7, wherein the PD-L1 binding antagonist
is selected from the group consisting of: YW243.55.S70, MPDL3280A,
MDX-1105, MEDI-4736, and MSB0010718C.
9. The combination of claim 2, wherein the PD-L/PD-1 Axis
antagonist is a PD-L2 binding antagonist.
10. The combination of claim 9, wherein the PD-L2 binding
antagonist is an antibody.
11. The combination of claim 9, wherein the PD-L2 binding
antagonist is an immunoadhesin.
12. The combination of any one of claim 1, wherein said
immunotherapeutics is a compound of any one of formula (I) to
(XIXb), or a pharmaceutically acceptable salt or solvate
thereof.
13. The combination of claim 1, further comprising an effective
amount of an an anticancer agent.
14. The combination of claim 13, wherein said anticancer agent is
an antimetabolite, an inhibitor of topoisomerase I and II, an
alkylating agent, a microtubule inhibitor, an antiandrogen agent, a
GNRh modulator or mixtures thereof.
15. The combination of claim 13, wherein said anticancer agent is a
chemotherapeutic agent selected from the group consisting of
tamoxifen, raloxifene, anastrozole, exemestane, letrozole,
imatanib, paclitaxel, cyclophosphamide, lovastatin, minosine,
gemcitabine, cytarabine, 5-fluorouracil, methotrexate, docetaxel,
goserelin, vincristine, vinblastine, nocodazole, teniposide
etoposide, gemcitabine, epothilone, vinorelbine, camptothecin,
daunorubicin, actinomycin D, mitoxantrone, acridine, doxorubicin,
epirubicin, or idarubicin.
16. The combination of claim 1, wherein said immunotherapeutic is
of an amount that is capable of: (1) inducing IFN-.alpha. in an
enriched human blood DCs; (2) inducing TNF-.alpha. in an enriched
human blood DCs; (3) inducing IL-12-.alpha. in an enriched human
blood DCs; (4) activating CD45+ immune cells in tumor
microenvironment; (5) activating CD4+ and CD8+ T cells in tumor
microenvironment; (6) activating NK cells in tumor
microenvironment; (7) activating plasmacytoid dendritic cells (pDC)
and myeloid dendritic cells (mDc) in tumor microenvironment; (8)
activating macrophages and Monocytes in tumor microenvironment;
and/or (9) increasing migratory DCs in draining lymph nodes.
17. A kit, comprising the combination of claim 1.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation of U.S. patent
application Ser. No. 15/401,843, filed Jan. 9, 2017, which is a
continuation-in-part of PCT/CN2015/083585, filed Jul. 8, 2015,
which claims the benefit of, and priority to, Chinese Patent
Application Serial Nos. 201410325480.9, filed Jul. 9, 2014, and
201410440824.0, filed Sep. 1, 2014, the entire disclosures of which
are hereby incorporated by reference in their entireties.
FIELD OF THE INVENTION
[0002] The present invention relates to therapeutic combinations
and methods for treating cancers using combination therapy.
BACKGROUND OF THE INVENTION
[0003] Therapeutic antibodies have been used in clinical
applications for over twenty years. Currently, there are fifteen
anti-tumor antibody drugs in clinic, including Rituxan (1997),
Herceptin (1998), Mylotarg (2000), Campath (2001), Zevalin (2002),
Bexxer (2003), Avastin (2004), Erbitux (2004), Vectibix (2006),
Arzerra (2009); Benlysta (2011); Yervoy (2011), Adcetris (2011),
Perjeta (2012), and Kadcyla (2013). These antibodies target mainly
four molecules: EGFR, Her2, CD20 and VEGF.
[0004] In general, therapeutic antibodies kill tumor cells via
three mechanisms (Scott A M, Wolchok J D, Old L J. Antibody therapy
of cancer. Nat Rev Cancer. (2012), 12:278-87): (1) Direct antibody
action, that is, blockade or agonist activity of ligand/receptor
signaling, induction of apoptosis, and delivery of drugs or
cytotoxic agents. Antibody receptor activation activity can produce
direct tumor cell killing effect. For example, some antibodies can
bind to receptors on the surface of tumor cells, activate the
receptor, leading to apoptosis (e.g., in mitochondria). Antibodies
can also mediate tumor cell killing by receptor-antagonistic
activity. For example, certain antibodies can bind to cell surface
receptors and block dimerization, kinase activation and downstream
signaling, thereby inhibiting proliferation and promote apoptosis.
Binding of antibodies to an enzyme can lead to neutralization,
signal abrogation, and cell death. (2) Through immune-mediated cell
killing mechanisms include complement-dependent cytotoxicity (CDC),
antibody-dependent cell-mediated cytotoxicity (ADCC), T cell
function regulation, etc. Immune-mediated killing of tumor cells
can be accomplished through the following ways: induction of
phagocytosis, complement activation, antibody-dependent
cell-mediated cytotoxicity, genetically-modified T cells being
targeted to the tumor by single-chain variable fragment (scFv),
through antibody-mediated antigenic cross presentation to dendritic
cell to activate T cells, inhibition of T cell inhibitory
receptors, such as cytotoxic T lymphocyte-associated antigen 4
(CTLA4). Of them, the Fc portion of the antibody feature is
especially important for CDC and ADCC-mediated tumor cell killing
effect. (3) Specific effect of antibody on tumor vasculature and
matrix, through trapping of vascular receptor antagonist or ligand
to induce vascular and stromal cells ablation, including: stromal
cell inhibition, delivery of toxins to stromal cells, and delivery
of toxins to the vasculature. (Scott A M, Wolchok J D, Old L J.
Antibody therapy of cancer. Nat Rev Cancer. 2012, 12
(4):278-87).
[0005] Therapeutic monoclonal antibody drugs have advanced
anti-cancer drug research and development. However, some issues
still need further study to be solved, such as antibody
immunogenicity, tolerance of long-term use of tumor target, and
long-term effects of simple single blockade of signal transduction
pathway. In short, a simple majority of antibodies are difficult to
achieve long-term efficient inhibition and killing of tumor
cells.
[0006] Antibody-drug conjugates combine targeting function and
small molecule drug with particular pharmacokinetics. The structure
of antibody-drug conjugates is the attachment of a monoclonal
antibody with targeting function to a compound with specific
pharmacological properties. This technique requires the therapeutic
antibody have binding specificity to a target, to be coupled to a
molecule with therapeutic effect or other functions such as
cyto-toxins. Many factors affect the effect of this type of
antibodies, such as endocytosis of the coupled antibody, stability
of the coupling, and release and killing activity of the
toxins.
[0007] Antibodies-drug conjugates have direct and indirect
anti-cancer effect. The antibody blocks or activates
ligand/receptor signaling, induces apoptosis, and at the same time
can present or deliver payload drug directly or indirectly (such as
a drug, toxin, small interfering RNA or radioisotope) to the tumor
cells. Therapeutic antibody drug conjugate utilizes dual
characteristics of the antibody and the coupled drug, first is the
binding function that it specifically binds to the target molecule,
second is the tumor cell killing function of the antibody itself,
and the third is the particular effect of the conjugated drug.
Current antibody-drug conjugates drugs are limited in how to kill
tumor cells directly. However, because of the tough requirement of
technologies in antibody, linker molecule, toxin molecules, and
conjugation, as well as the limitation of bringing toxins within
the tumor microenvironment molecules, there are still some
difficulties in actual clinical studies.
SUMMARY OF THE INVENTION
[0008] In one aspect, the present invention provides a therapeutic
combination, comprising: (i) an effective amount of a PD-L/PD-1
Axis antagonist; and (ii) an effective amount of an
immunotherapeutic that is capable of activating a human
plasmacytoid dendritic cell, myeloid dendritic cell, or NK cell, or
a combination thereof.
[0009] In some embodiments, the PD-L/PD-1 Axis antagonist is
selected from the group consisting of a PD-1 binding antagonist, a
PD-L1 binding antagonist and a PD-L2 binding antagonist.
[0010] In some embodiments, the PD-L/PD-1 Axis antagonist is a PD-1
binding antagonist.
[0011] In some embodiments, the PD-1 binding antagonist inhibits
the binding of PD-1 to its ligand binding partners.
[0012] In some embodiments, the PD-1 binding antagonist inhibits
the binding of PD-1 to PD-L1.
[0013] In some embodiments, the PD-1 binding antagonist inhibits
the binding of PD-1 to PD-L2.
[0014] In some embodiments, the PD-1 binding antagonist inhibits
the binding of PD-1 to both PD-L1 and PD-L2.
[0015] In some embodiments, the PD-1 binding antagonist is an
antibody, such as MDX-1106, Merck 3745, CT-011, AMP-224 or
AMP-514.
[0016] In some embodiments, the PD-L/PD-1 Axis antagonist is a
PD-L1 binding antagonist.
[0017] In some embodiments, the PD-L1 binding antagonist inhibits
the binding of PD-L1 to PD-1.
[0018] In some embodiments, the PD-L1 binding antagonist inhibits
the binding of PD-L1 to B7-1.
[0019] In some embodiments, the PD-L1 binding antagonist inhibits
the binding of PD-L1 to both PD-1 and B7-1.
[0020] In some embodiments, the PD-L1 binding antagonist is an
antibody, such as one selected from the group consisting of:
YW243.55.S70, MPDL3280A, MDX-1105, MEDI-4736, and MSB0010718C.
[0021] In some embodiments, the PD-L/PD-1 Axis antagonist is a
PD-L2 binding antagonist.
[0022] In some embodiments, the PD-L2 binding antagonist is an
antibody.
[0023] In some embodiments, the PD-L2 binding antagonist is an
immunoadhesin.
[0024] In some embodiments, the treatment results in a sustained
response in the individual after cessation of the treatment.
[0025] In some embodiments, the immunotherapeutic is administered
continuously, intermittently.
[0026] In some embodiments, the immunotherapeutic is administered
before the PD-L/PD-1 Axis antagonist.
[0027] In some embodiments, the immunotherapeutic is administered
simultaneous with the PD-L/PD-1 Axis antagonist.
[0028] In some embodiments, the immunotherapeutic is administered
after the PD-L/PD-1 Axis antagonist.
[0029] In some embodiments, the individual has colorectal cancer,
melanoma, non-small cell lung cancer, ovarian cancer, breast
cancer, pancreatic cancer, a hematological malignancy or renal cell
carcinoma.
[0030] In some embodiments, wherein the PD-L/PD-1 Axis antagonist
is administered intravenously, intramuscularly, subcutaneously,
topically, orally, transdermally, intraperitoneally,
intraorbitally, by implantation, by inhalation, intrathecally,
intraventricularly, or intranasally.
[0031] In some embodiments, the immunotherapeutic is capable of
binding specifically to human TLR7 and/or TLR8.
[0032] In some embodiments, the immunotherapeutic comprises: (a)
single-stranded RNA (ssRNA), preferably ORN02, ORN06, ssPoly(U),
ssRNA40, ssRNA41, ssRNA-DR, or Poly(dT); or (b) a receptor ligand
analogs, preferably CL075, CL097, CL264, CL307, Gardiquimod,
Loxoribine, Imiquimod, or Resiquimod.
[0033] In some embodiments, the immunotherapeutics is a compound of
any one of formula (I) to (XIXb), or a pharmaceutically acceptable
salt or solvate thereof.
[0034] In some embodiments, the immunotherapeutic has a structure
of Formula (I):
##STR00001##
wherein dashed line represents bond or absence of bond: X is S or
--NR.sub.1, R.sub.1 is -W.sub.0-W.sub.1-W.sub.2-W.sub.3-W.sub.4.
W.sub.0 is a bond, alkyl, alkenyl, alkynyl, alkoxy, or
-alkyl-S-alkyl-, W.sub.1 is a bond, --O--, or --NR.sub.2--, wherein
R.sub.2 is hydrogen, alkyl or alkenyl, W.sub.2 is a bond, --O--,
--C(O)--, --C(S)--, or --S(O).sub.2--, W.sub.3 is a bond,
--NR.sub.3--, wherein R.sub.3 is hydrogen, alkyl or alkenyl,
W.sub.4 is hydrogen, alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl,
aryl, aryloxy, heteroaryl, or heterocyclyl, each of which is
optionally substituted by one or more substituents selected from
the group consisting of hydroxyl, alkoxy, alkyl, alkenyl, alkynyl,
cycloalkyl, aryl, heteroaryl, heterocyclyl, --NH.sub.2, nitro,
-alkyl-hydroxyl, -alkyl-aryl, -alkyl-heteroaryl,
-alkyl-heterocyclyl, --O--R.sub.4, --O-alkyl-R.sub.4,
-alkyl-O--R.sub.4, --C(O)--R.sub.4, -alkyl-C(O)--R.sub.4,
-alkyl-C(O)--O--R.sub.4, --C(O)--O--R.sub.4, --S--R.sub.4,
--S(O).sub.2--R.sub.4, --NH--S(O).sub.2--R.sub.4,
-alkyl-S--R.sub.4, -alkyl-S(O).sub.2--R.sub.4, --NHR.sub.4,
--NR.sub.4R.sub.4, --NH-alkyl-R.sub.4, halogen, --CN, --NO.sub.2,
and --SH, wherein R.sub.4 is independently hydrogen, alkyl,
alkenyl, -alkyl-hydroxyl, aryl, heteroaryl, heterocyclyl, or
haloalkyl; Z is hydrogen, alkyl, alkenyl, alkynyl, alkoxy, aryl,
haloalkyl, heteroaryl, heterocyclyl, each of which can be
optionally substituted by one or more substituents selected from
the group consisting of hydroxyl, alkoxy, alkyl, alkenyl, alkynyl,
aryl, heteroaryl, heterocyclyl, halogen, cyano, nitro,
--N(R.sub.5).sub.2, -alkoxy-alkyl, -alkoxy-alkenyl, --C(O)-alkyl,
--C(O)--O-alkyl, --O--C(O)-alkyl, --C(O)--N(R.sub.5).sub.2, aryl,
heteroaryl, --CO-aryl, and --CO-heteroaryl, wherein each R.sub.5 is
independently hydrogen, alkyl, haloalkyl, -alkyl-aryl, or
-alkyl-heteroaryl; R is hydrogen, alkyl, alkoxy, haloalkyl,
halogen, aryl, heteroaryl, heterocyclyl, each of which is
optionally substituted by one or more substituents selected from
the group consisting of hydroxyl, alkoxy, alkyl, alkenyl, alkynyl,
cycloalkyl, aryl, heteroaryl, heterocyclyl, --NH.sub.2, nitro,
-alkyl-hydroxyl, -alkyl-aryl, -alkyl-heteroaryl,
-alkyl-heterocyclyl, --O--R.sub.4, --O-alkyl-R.sub.4,
-alkyl-O--R.sub.4, --C(O)--R.sub.4, --C(O)--NH--R.sub.4,
--C(O)--NR.sub.4R.sub.4, -alkyl-C(O)--R.sub.4,
-alkyl-C(O)--O--R.sub.4, --C(O)--O--R.sub.4, --O--C(O)--R.sub.4,
--S--R.sub.4, --C(O)--S--R.sub.4, --S--C(O)--R.sub.4,
--S(O).sub.2--R.sub.4, --NH--S(O).sub.2--R.sub.4,
-alkyl-S--R.sub.4, -alkyl-S(O).sub.2--R.sub.4, --NHR.sub.4,
--NR.sub.4R.sub.4, --NH-alkyl-R.sub.4, halogen, --CN, and --SH,
wherein R.sub.4 is independently hydrogen, alkyl, alkenyl, alkoxy,
-alkyl-hydroxyl, aryl, heteroaryl, heterocyclyl, or haloalkyl; n is
0, 1, 2, 3, or 4; Y is --NR.sub.6R.sub.7, --CR.sub.6R.sub.7R.sub.8,
or -alkyl-NH.sub.2, each of which can be optionally substituted by
one or more substituents selected from the group consisting of
hydroxyl, alkoxy, alkyl, alkenyl, alkynyl, --NH.sub.2, halogen,
--N(R.sub.5).sub.2, -alkoxyalkyl, -alkoxy-alkenyl, --C(O)-alkyl,
--C(O)--O-alkyl, --C(O)--N(R.sub.5).sub.2, aryl, heteroaryl,
--CO-aryl, and --CO-heteroaryl. wherein R.sub.6, R.sub.7 and
R.sub.8 are independently hydrogen, alkyl, alkenyl, alkoxy,
alkylamino, dialkylamino, alkylthio, arylthio, -alkyl-hydroxyl,
-alkyl-C(O)--O--R.sub.9, -alkyl-C(O)--R.sub.9, or
-alkyl-O--C(O)--R.sub.9, wherein each R.sub.5 is independently
hydrogen, alkyl, haloalkyl, -alkyl-aryl, or -alkyl-heteroaryl,
wherein R.sub.9 is hydrogen, alkyl, alkenyl, halogen, or haloalkyl;
and X and Z taken together may optionally form a (5-9)-membered
ring.
[0035] In some embodiments, the immunotherapeutic is a compound
selected from the group consisting of:
2-propylthiazolo[4,5-c]quinolin-4-amine,
1-(2-methylpropyl)-1H-imidazo[4,5-c]quinolin-4-amine,
4-amino-2-(ethoxymethyl)-a,a-di-methyl-1H-imidazo[4,5-c]quinoline-1-ethan-
ol,
1-(4-amino-2-ethylaminomethylimidazo-[4,5-c]quinolin-1-yl)-2-methylpro-
pan-2-ol,
N-[4-(4-amino-2-ethyl-1H-imidazo[4,5-c]quinolin-1-yl)butyl-]meth-
anesulfonamide,
4-amino-2-ethoxymethyl-aa-dimethyl-6,7,8,9-tetrahydro-1h-imidazo[4,5-c]qu-
inoline-1-ethanol,
4-amino-aa-dimethyl-2-methoxyethyl-1h-imidazo[4,5-c]quinoline-1-ethanol,
1-{2-[3-(benzyloxy)propoxy]ethyl}-2-(ethoxymethyl)-1H-imidazo[4,5-c]quino-
lin-4-amine,
N-[4-(4-amino-2-butyl-1H-imidazo[4,5-c][1,5]naphthyridin-1-yl)butyl]-n'-b-
utylurea,
N1-[2-(4-amino-2-butyl-1H-imidazo[4,5-c][1,5]naphthyridin-1-yl)e-
thyl]-2-amino-4-methylpentanamide,
N-(2-{2-[4-amino-2-(2-methoxyethyl)-1H-imidazo[4,5-c]quinolin-1-yl]ethoxy-
}ethyl)-n'-phenylurea,
1-(2-amino-2-methylpropyl)-2-(ethoxymethyl)-1H-imidazo[4,5-c]quinolin-4-a-
mine,
1-{4-[(3,5-dichlorophenyl)sulfonyl]butyl}-2-ethyl-1H-imidazo[4,5-c]q-
uinolin-4-amine,
N-(2-{2-[4-amino-2-(ethoxymethyl)-1H-imidazo[4,5-c]quinolin-1-yl]ethoxy}e-
thyl)-n'-cyclohexylurea,
N-{3-[4-amino-2-(ethoxymethyl)-1H-imidazo[4,5-c]quinolin-1-yl]propyl}-n'--
(3-cyanophenyl)thiourea,
N-[3-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)-2,2-dimethylpropyl]-
benzamide,
2-butyl-1-[3-(methylsulfonyl)propyl]-1H-imidazo[4,5-c]quinolin--
4-amine,
N-{2-[4-amino-2-(ethoxymethyl)-1H-imidazo[4,5-c]quinolin-1-yl]-1,-
1-dimethylethyl}-2-ethoxyacetamide,
1-[4-amino-2-ethoxymethyl-7-(pyridin-4-yl)-1H-imidazo[4,5-c]quinolin-1-yl-
]-2-methylpropan-2-ol,
1-[4-amino-2-(ethoxymethyl)-7-(pyridin-3-yl)-1H-imidazo[4,5-c]quinolin-1--
yl]-2-methylpropan-2-ol,
N-{3-[4-amino-1-(2-hydroxy-2-methylpropyl)-2-(methoxyethyl)-1H-imidazo[4,-
5-c]quinolin-7-yl]phenyl}methanesulfonamide,
1-[4-amino-7-(5-hydroxymethylpyridin-3-yl)-2-(2-methoxyethyl)-1H-imidazo[-
4,5-c]quinolin-1-yl]-2-methylpropan-2-ol,
3-[4-amino-2-(ethoxymethyl)-7-(pyridin-3-yl)-1H-imidazo[4,5-c]quinolin-1--
yl]propane-1,2-diol,
1-[2-(4-amino-2-ethoxymethyl-1H-imidazo[4,5-c]quinolin-1-yl)-1,1-dimethyl-
ethyl]-3-propylurea,
1-[2-(4-amino-2-ethoxymethyl-1H-imidazo[4,5-c]quinolin-1-yl)-1,1-dimethyl-
ethyl]-3-cyclopentylurea,
1-[(2,2-dimethyl-1,3-dioxolan-4-yl)methyl]-2-(ethoxymethyl)-7-(4-hydroxym-
ethylphenyl)-1H-imidazo[4,5-c]quinolin-4-amine,
4-[4-amino-2-ethoxymethyl-1-(2-hydroxy-2-methylpropyl)-1H-imidazo[4,5-c]q-
uinolin-7-yl]-N-methoxy-N-methylbenzamide,
2-ethoxymethyl-N1-isopropyl-6,7,8,9-tetrahydro-1H-imidazo[4,5-c]quinoline-
-1,4-diamine,
1-[4-amino-2-ethyl-7-(pyridin-4-yl)-1H-imidazo[4,5-c]quinolin-1-yl]-2-met-
hylpropan-2-ol,
N-[4-(4-amino-2-ethyl-1H-imidazo[4,5-c]quinolin-1-yl)butyl]methanesulfona-
mide, and
N-[4-(4-amino-2-butyl-1H-imidazo[4,5-c][1,5]naphthyridin-1-yl)bu-
tyl]-n'-cyclohexylurea.
[0036] In some embodiments, the immunotherapeutic has a structure
of Formula (II):
##STR00002##
wherein V is --NR.sub.6R.sub.7, wherein each of R.sub.6 and R.sub.7
is independently hydrogen, alkyl, alkenyl, alkoxy, alkylamino,
dialkylamino, alkylthio, arylthio, -alkyl-hydroxyl,
-alkyl-C(O)--O--R.sub.9, -alkyl-C(O)--R.sub.9, or
-alkyl-O--C(O)--R.sub.9, wherein R.sub.9 is hydrogen, alkyl,
alkenyl, halogen, or haloalkyl; R.sub.10 and R.sub.11 are
independently hydrogen, alkyl, alkenyl, aryl, haloalkyl,
heteroaryl, heterocyclyl, or cycloalkyl, each of which is
optionally substituted by one or more substituents selected from
the group consisting of hydroxyl, alkoxy, alkyl, alkenyl, alkynyl,
halogen, --N(R.sub.5).sub.2, -alkoxy-alkyl, -alkoxy-alkenyl,
--C(O)-alkyl, --C(O)--O-alkyl, --C(O)--N(R.sub.5).sub.2, aryl,
heteroaryl, --CO-aryl, and --CO-heteroaryl, wherein each R.sub.5 is
independently hydrogen, alkyl, haloalkyl, -alkyl-aryl, or
-alkyl-heteroaryl.
[0037] In some embodiments, the therapeutic combination or
pharmaceutical composition of the present invention further
comprises an effective amount of an additional therapeutic agent,
such as an anticancer agent.
[0038] In some embodiments, the anticancer agent is an
antimetabolite, an inhibitor of topoisomerase I and II, an
alkylating agent, a microtubule inhibitor, an antiandrogen agent, a
GNRh modulator or mixtures thereof.
[0039] In some embodiments, the additional therapeutic agent is a
chemotherapeutic agent selected from the group consisting of
tamoxifen, raloxifene, anastrozole, exemestane, letrozole,
imatanib, paclitaxel, cyclophosphamide, lovastatin, minosine,
gemcitabine, cytarabine, 5-fluorouracil, methotrexate, docetaxel,
goserelin, vincristine, vinblastine, nocodazole, teniposide
etoposide, gemcitabine, epothilone, vinorelbine, camptothecin,
daunorubicin, actinomycin D, mitoxantrone, acridine, doxorubicin,
epirubicin, or idarubicin.
[0040] In another aspect, the present invention provides a method
for treating a disease condition in a subject that is in need of
such treatment, comprising administering to the subject the
therapeutic combination or pharmaceutical composition provided
herein.
[0041] In some embodiments, the diseases condition is tumor. In
some embodiments, the disease condition comprises abnormal cell
proliferation.
[0042] In some embodiments, the abnormal cell proliferation
comprises a pre-cancerous lesion. In some embodiments, the abnormal
proliferation is of cancer cells.
[0043] In some embodiments, the cancer is selected from the group
consisting of: breast cancer, colorectal cancer, diffuse large
B-cell lymphoma, endometrial cancer, follicular lymphoma, gastric
cancer, glioblastoma, head and neck cancer, hepatocellular cancer,
lung cancer, melanoma, multiple myeloma, ovarian cancer, pancreatic
cancer, prostate cancer, and renal cell carcinoma.
[0044] In some embodiments, the immunotherapeutic is of an amount
that is capable of: [0045] (1) inducing IFN-.alpha. in an enriched
human blood DCs; [0046] (2) inducing TNF-.alpha. in an enriched
human blood DCs; [0047] (3) inducing IL-12-.alpha. in an enriched
human blood DCs; [0048] (4) activating CD45+ immune cells in tumor
microenvironment; [0049] (5) activating CD4+ and CD8+ T cells in
tumor microenvironment; [0050] (6) activating NK cells in tumor
microenvironment; [0051] (7) activating plasmacytoid dendritic
cells (pDC) and myeloid dendritic cells (mDc) in tumor
microenvironment; [0052] (8) activating macrophages and Monocytes
in tumor microenvironment; and/or [0053] (9) increasing migratory
DCs in draining lymph nodes.
[0054] In some embodiments, the method comprises administering to
the subject an oral formulation comprising the immunotherapeutic
(such as R848 and its analogues) in a dose of between about 0.0005
mg/kg, 0.0006 mg/kg, 0.0007 mg/kg, 0.0008 mg/kg, 0.0009 mg/kg,
0.001 mg/kg, 0.002 mg/kg, 0.003 mg/kg, 0.004 mg/kg, 0.005 mg/kg,
0.006 mg/kg, 0.007 mg/kg, 0.008 mg/kg, 0.009 mg/kg, or 0.01 mg/kg,
to about 0.02 mg/kg, twice per week.
[0055] In some embodiments, the method comprises administering to
the subject an oral formulation comprising the immunotherapeutic
(such as R848 and its analogues) in a dose of less than or about
0.005 mg/kg, 0.006 mg/kg, 0.007 mg/kg, 0.008 mg/kg, 0.009 mg/kg, or
0.01 mg/kg, twice per week.
[0056] In some embodiments, the method comprises administering to
said subject an intravenous formulation comprising said
immunotherapeutic (such as R848 and its analogues) in a dose of
between about 0.0005 mg/kg, 0.0006 mg/kg, 0.0007 mg/kg, 0.0008
mg/kg, 0.0009 mg/kg, 0.001 mg/kg, 0,002 mg/kg, 0,003 mg/kg, 0,004
mg/kg, 0,005 mg/kg, or 0.006 mg/kg to about 0.015 mg/kg, weekly. In
some embodiments, the method comprises administering to said
subject an intravenous formulation comprising said
immunotherapeutic (such as R848 and its analogues) in a dose of
between about 0.0008 mg/kg to about 0.0067 mg/kg, weekly.
[0057] In some embodiments, the method comprises administering to
said subject an intravenous formulation comprising said
immunotherapeutic in a dose of less than or about 0,003 mg/kg,
0,004 mg/kg, 0,005 mg/kg, or 0.006 mg/kg to about 0.007 mg/kg,
weekly.
[0058] In some embodiments, the immunotherapeutic in the subject
has a local concentration that is between about 0.005 .mu.g/ml to
about 12 .mu.g/ml.
[0059] In some embodiments, the immunotherapeutic in the subject
has a local concentration that is is between about 0.05 .mu.g/ml,
0.1 .mu.g/ml, 0.15 .mu.g/ml, 0.2 .mu.g/ml, 0.3 .mu.g/ml, or 0.4
.mu.g/ml, to about 0.5 .mu.g/ml.
[0060] In a further aspect, the present invention provides a kit
that contains the therapeutic combination provided herein, and
optionally with an instruction.
BRIEF DESCRIPTION OF THE DRAWINGS
[0061] The novel features of the invention are set forth with
particularity in the appended claims. A better understanding of the
features and advantages of the present invention will be obtained
by reference to the following detailed description that sets forth
illustrative embodiments, in which the principles of the invention
are utilized, and the accompanying drawings of which:
[0062] FIG. 1 depicts effects of anti-PDL1 mAb treatment on SCCVII
tumour growth. SCCVII Tumours were inoculated as described in the
Materials and methods. Tumour-inoculated mice received an
intraperitoneal injection of control rat immunoglobulin or
anti-PDL1 mAb (200 ug/mouse) combination with TLRL three times a
week. The mean tumour volume.+-.SD was determined in each group of
five to eight mice.
[0063] FIG. 2 depicts effects of anti-PDL1 mAb treatment on CT26
tumour growth. CT26 Tumours were inoculated as described in the
Materials and methods. Tumour-inoculated mice received an
intraperitoneal injection of control rat immunoglobulin or
anti-PDL1 mAb (200 ug/mouse) combination with TLRL three times a
week. The mean tumour volume.+-.SD was determined in each group of
five to eight mice.
[0064] FIGS. 3A-3G depict analysis of cytokine production by
enriched human DCs from three healthy donors. Enriched human DCs
were plated in a 96-well plate and cultured with allogeneic
untreated (medium) or treated different concentration of TLRL
directly for 20-22 h in 37.degree. C. incubator. The supernatants
were collected and human IFN-.alpha., IL-12(p70) and TNF-.alpha.
were analyzed by ELISA. Data are given as mean.+-.SD of triplicate
cultures. Three independent experiments from three healthy donors
were performed. A: TLRL induced IFN-.alpha. expression in enriched
human blood DCs (CD3+/CD19+/CD14+/CD16+) from donor 1. B: TLRL
induced IFN-.alpha. expression in enriched human blood DCs in
Experiment #2 from donor 2. C: TLRL induced TNF-.alpha. expression
in enriched human blood DCs in Experiment #2 from donor 2. D: TLRL
induced IL-12 expression in enriched human blood DCs in Experiment
#2 from donor 2. E: TLRL induced IFN-.alpha. expression in enriched
human blood DCs Experiment #3 from donor 3. F: TLRL induced
TNF-.alpha. expression in enriched human blood DCs in Experiment #3
from donor 3. G: TLRL induced IL-12 expression in enriched human
blood DCs in Experiment #3 from donor 3.
[0065] FIGS. 4A-C depicts expression of IFN inducible genes in
mouse PBMC after TLRL injection. RNA was isolated from PBMCs
cryopreserved with TRIzol reagent at variable time points and
Relative expression of IFN inducible genes were determined by
quantitative RT-PCR. MX2 gene was detected over time course of 5
hours post TLRL injection (4A) and MX2 and ISG15 genes were
measured with various dose of TLRL at 2 hours post injection (4B)
and (4C), respectively. Values indicate the mRNA expression of
indicated IFN inducible genes relative to housekeeping gene Actin.
Bar graphs represent data from 3 individual animals. **P<0.01;
***P<0.001.
[0066] FIG. 5 depicts tumor volumes of mice treated with anti-PD1
alone. R848 alone, and combination of cnti-PD1 and R848 against
couse colon cells C26 in BALB/c mice. Tumor volumes are shown as
the mean+SEM of 10 mice per group. Mice treated with the
combination of ANTI-PD-1 and R848 intravenously had the best
antitumor response with 96.15% (p<0.01) tumor inhibition.
[0067] FIG. 6 depicts Tumor columes of mice treated with anti-PD-L1
alone, R848 alone, and combination of anti-PD-L1 and R848 against
aouse colon cells C26 in BALB/c mice. Tumor volumes are shown as
the mean+SEM of 10 mice per group. Mice treated with the
combination of Anti-PD-L1 and R848 intravenously had the best
antitumor response with 96.15% (p<0.01) tumor inhibition.
DETAILED DESCRIPTION OF THE INVENTION
[0068] Several aspects of the invention are described below with
reference to example applications for illustration. It should be
understood that numerous specific details, relationships, and
methods are set forth to provide a full understanding of the
invention. One having ordinary skill in the relevant art, however,
will readily recognize that the invention can be practiced without
one or more of the specific details or with other methods. The
present invention is not limited by the illustrated ordering of
acts or events, as some acts may occur in different orders and/or
concurrently with other acts or events.
[0069] Furthermore, not all illustrated acts or events are required
to implement a methodology in accordance with the present
invention.
[0070] The terminology used herein is for the purpose of describing
particular embodiments only and is not intended to be limiting of
the invention. As used herein, the singular forms "a", "an" and
"the" are intended to include the plural forms as well, unless the
context clearly indicates otherwise. Furthermore, to the extent
that the terms "including", "includes", "having", "has", "with", or
variants thereof are used in either the detailed description and/or
the claims, such terms are intended to be inclusive in a manner
similar to the term "comprising".
[0071] The term "about" or "approximately" means within an
acceptable error range for the particular value as determined by
one of ordinary skill in the art, which will depend in part on how
the value is measured or determined, i.e., the limitations of the
measurement system. For example, "about" can mean within 1 or more
than 1 standard deviation, per the practice in the art.
Alternatively, "about" can mean a range of up to 20%, preferably up
to 10% more preferably up to 5%, and more preferably still up to 1%
of a given value. Alternatively, particularly with respect to
biological systems or processes, the term can mean within an order
of magnitude, preferably within 5-fold, and more preferably within
2-fold, of a value. Where particular values are described in the
application and claims, unless otherwise stated the term "about"
meaning within an acceptable error range for the particular value
should be assumed.
I. Definitions and Abbreviations
[0072] Unless defined otherwise, all technical and scientific terms
used herein generally have the same meaning as commonly understood
by one of ordinary skill in the art to which this invention
belongs. Generally, the nomenclature used herein and the laboratory
procedures in cell culture, molecular genetics, organic chemistry
and nucleic acid chemistry and hybridization are those well-known
and commonly employed in the art. Standard techniques are used for
nucleic acid and peptide synthesis. The techniques and procedures
are generally performed according to conventional methods in the
art and various general references, which are provided throughout
this document. The nomenclature used herein and the laboratory
procedures in analytical chemistry, and organic synthetic described
below are diose well-known and commonly employed in the art.
Standard techniques, or modifications thereof, are used for
chemical syntheses and chemical analyses.
[0073] The term "alkyl," by itself or as part of another
substituent, means, unless otherwise stated, a straight or branched
chain, or cyclic hydrocarbon radical, or combination thereof, which
may be fully saturated, mono- or polyunsaturated and can include
di- and multivalent radicals, having the number of carbon atoms
designated (i.e. C.sub.1-C.sub.10 means one to ten carbons).
Examples of saturated hydrocarbon radicals include, but are not
limited to, groups such as methyl, ethyl, n-propyl, isopropyl,
n-butyl, t-butyl, isobutyl, sec-butyl, cyclohexyl,
(cyclohexyl)methyl, cyclopropylmethyl, homologs and isomers of, for
example, n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like. An
unsaturated alkyl group is one having one or more double bonds or
triple bonds. Examples of unsaturated alkyl groups include, but are
not limited to, vinyl, 2-propenyl, crotyl, 2-isopentenyl,
2-(butadienyl), 2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1-
and 3-propynyl, 3-butynyl, and the higher homologs and isomers. The
term "alkyl," unless otherwise noted, is also meant to include
those derivatives of alkyl defined in more detail below, such as
"heteroalkyl." Alkyl groups, which are limited to hydrocarbon
groups, are termed "homoalkyl".
[0074] The term "alkylene" by itself or as part of another
substituent means a divalent radical derived from an alkane, as
exemplified, but not limited, by
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2--, and further includes those
groups described below as "heteroalkylene." Typically, an alkyl (or
alkylene) group will have from 1 to 24 carbon atoms, with those
groups having 10 or fewer carbon atoms being preferred in the
present invention. A "lower alkyl" or "lower alkylene" is a shorter
chain alkyl or alkylene group, generally having eight or fewer
carbon atoms.
[0075] The terms "alkoxy," "alkylamino" and "alkylthio" (or
thioalkoxy) are used in their conventional sense, and refer to
those alkyl groups attached to the remainder of the molecule via an
oxygen atom, an amino group, or a sulfur atom, respectively.
[0076] The term "heteroalkyl," by itself or in combination with
another term, means, unless otherwise stated, a stable straight or
branched chain, or cyclic hydrocarbon radical, or combinations
thereof, consisting of the stated number of carbon atoms and at
least one heteroatom selected from the group consisting of O, N, Si
and S, and wherein the nitrogen and sulfur atoms may optionally be
oxidized and the nitrogen heteroatom may optionally be quaternized.
The heteroatom(s) O, N and S and Si may be placed at any interior
position of the heteroalkyl group or at the position at which the
alkyl group is attached to the remainder of the molecule. Examples
include, but are not limited to, --CH.sub.2--CH.sub.2--O--CH.sub.3,
--CH.sub.2--CH.sub.2--NH--CH.sub.3,
--CH.sub.2--CH.sub.2--N(CH.sub.3)--CH.sub.3,
--CH.sub.2--S--CH.sub.2--CH.sub.3, --CH.sub.2--CH.sub.2,
--S(O)--CH.sub.3, --CH.sub.2--CH.sub.2--S(O).sub.2--CH.sub.3,
--CH.dbd.CH--O--CH.sub.3, --Si(CH.sub.3).sub.3,
--CH.sub.2--CH.dbd.N--OCH.sub.3, and
--CH.dbd.CH--N(CH.sub.3)--CH.sub.3. Up to two heteroatoms may be
consecutive, such as, for example, --CH.sub.2--NH--OCH.sub.3 and
--CH.sub.2--O--Si(CH.sub.3).sub.3. Similarly, the term
"heteroalkylene" by itself or as part of another substituent means
a divalent radical derived from heteroalkyl, as exemplified, but
not limited by, --CH.sub.2--CH.sub.2--S--CH.sub.2--CH.sub.2-- and
--CH.sub.2--S--CH.sub.2--CH.sub.2--NH--CH.sub.2--. For
heteroalkylene groups, heteroatoms can also occupy either or both
of the chain termini (e.g., alkyleneoxy, alkylenedioxy,
alkyleneamino, alkylenediamino, and the like). Still further, for
alkylene and heteroalkylene linking groups, no orientation of the
linking group is implied by the direction in which the formula of
the linking group is written. For example, the formula
--C(O).sub.2R'-- represents both --C(O).sub.2R'-- and
--R'C(O).sub.2--.
[0077] In general, an "acyl substituent" is also selected from the
group set forth above. As used herein, the term "acyl substituent"
refers to groups attached to, and fulfilling the valence of a
carbonyl carbon that is either directly or indirectly attached to
the polycyclic nucleus of the compounds of the present
invention.
[0078] The terms "cycloalkyl" and "heterocycloalkyl", by themselves
or in combination with other terms, represent, unless otherwise
stated, cyclic versions of "alkyl" and "heteroalkyl", respectively.
Additionally, for heterocycloalkyl, a heteroatom can occupy the
position at which the heterocycle is attached to the remainder of
the molecule. Examples of cycloalkyl include, but are not limited
to, cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl,
cycloheptyl, and the like. Examples of heterocycloalkyl include,
but are not limited to, 1-(1,2,5,6-tetrahydropyridyl),
1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-morpholinyl,
3-morpholinyl, tetrahydrofuran-2-yl, tetrahydrofuran-3-yl,
tetrahydrothien-2-yl, tetrahydrothien-3-yl, 1-piperazinyl,
2-piperazinyl, and the like.
[0079] The terms "halo" or "halogen," by themselves or as part of
another substituent, mean, unless otherwise stated, a fluorine,
chlorine, bromine, or iodine atom. Additionally, terms such as
"haloalkyl," are meant to include monohaloalkyl and polyhaloalkyl.
For example, the term "halo(C.sub.1-C.sub.4)alkyl" is mean to
include, but not be limited to, trifluoromethyl,
2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the
like.
[0080] As used herein, the term "haloalkyl" refers to an alkyl as
defined herein, that is substituted by one or more halo groups as
defined herein. Preferably the haloalkyl can be monohaloalkyl,
dihaloalkyl or polyhaloalkyl including perhaloalkyl. A
monohaloalkyl can have one iodo, bromo, chloro or fluoro within the
alkyl group. Dihaloalkyl and polyhaloalkyl groups can have two or
more of the same halo atoms or a combination of different halo
groups within the alkyl. Preferably, the polyhaloalkyl contains up
to 12, 10, or 8, or 6, or 4, or 3, or 2 halo groups. Non-limiting
examples of haloalkyl include fluoromethyl, difluoromethyl,
trifluoromethyl, chloromethyl, dichloromethyl, trichloromethyl,
pentafluoroethyl, heptafluoropropyl, difluorochloromethyl,
dichlorofluoromethyl, difluoroethyl, difluoropropyl, dichloroethyl
and dichloropropyl. A perhaloalkyl refers to an alkyl having all
hydrogen atoms replaced with halo atoms.
[0081] As used herein, the term "heteroaryl" refers to a 5-14
membered monocyclic- or bicyclic- or fused polycyclic-ring system,
having 1 to 8 heteroatoms selected from N, O, S or Se. Preferably,
the heteroaryl is a 5-10 membered ring system. Typical heteroaryl
groups include 2- or 3-thienyl, 2- or 3-furyl, 2- or 3-pyrrolyl,
2-, 4-, or 5-imidazolyl, 3-, 4-, or 5-pyrazolyl, 2-, 4-, or
5-thiazolyl, 3-, 4-, or 5-isothiazolyl, 2-, 4-, or 5-oxazolyl, 3-,
4-, or 5-isoxazolyl, 3- or 5-1,2,4-triazolyl, 4- or
5-1,2,3-triazolyl, tetrazolyl, 2-, 3-, or 4-pyridyl, 3- or
4-pyridazinyl, 3-, 4-, or 5-pyrazinyl, 2-pyrazinyl, 2-, 4-, or
5-pyrimidinyl.
[0082] The term "heteroaryl" also refers to a group in which a
heteroaromatic ring is fused to one or more aryl, cycloaliphatic,
or heterocycloalkyl rings, where the radical or point of attachment
is on the heteroaromatic ring. Nonlimiting examples include but are
not limited to 1-, 2-, 3-, 5-, 6-, 7-, or 8-indolizinyl, 1-, 3-,
4-, 5-, 6-, or 7-isoindolyl, 2-, 3-, 4-, 5-, 6-, or 7-indolyl, 2-,
3-, 4-, 5-, 6-, or 7-indazolyl, 2-, 4-, 5-, 6-, 7-, or 8-purinyl,
1-, 2-, 3-, 4-, 6-, 7-, 8-, or 9-quinolizinyl, 2-, 3-, 4-, 5-, 6-,
7-, or 8-quinoliyl, 1-, 3-, 4-, 5-, 6-, 7-, or 8-isoquinoliyl, 1-,
4-, 5-, 6-, 7-, or 8-phthalazinyl, 2-, 3-, 4-, 5-, or
6-naphthyridinyl, 2-, 3-, 5-, 6-, 7-, or 8-quinazolinyl, 3-, 4-,
5-, 6-, 7-, or 8-cinnolinyl, 2-, 4-, 6-, or 7-pteridinyl, 1-, 2-,
3-, 4-, 5-, 6-, 7-, or 8-4aH carbazolyl, 1-, 2-, 3-, 4-, 5-, 6-,
7-, or 8-carbzaolyl, 1-, 3-, 4-, 5-, 6-, 7-, 8-, or 9-carbolinyl,
1-, 2-, 3-, 4-, 6-, 7-, 8-, 9-, or 10-phenanthridinyl, 1-, 2-, 3-,
4-, 5-, 6-, 7-, 8-, or 9-acridinyl, 1-, 2-, 4-, 5-, 6-, 7-, 8-, or
9-perimidinyl, 2-, 3-, 4-, 5-, 6-, 8-, 9-, or 10-phenathrolinyl,
1-, 2-, 3-, 4-, 6-, 7-, 8-, or 9-phenazinyl, 1-, 2-, 3-, 4-, 6-,
7-, 8-, 9-, or 10-phenothiazinyl, 1-, 2-, 3-, 4- , 6-, 7-, 8-, 9-,
or 10-phenoxazinyl, 2-, 3-, 4-, 5-, 6-, or 1-, 3-, 4-, 5-, 6-, 7-,
8-, 9-, or 10-benzisoqinolinyl, 2-, 3-, 4-, or
5-thieno[2,3-b]furanyl, 2-, 3-, 5-, 6-, 7-, 8-, 9-, 10-, or
11-7H-pyrazino[2,3-c]carbazolyl, 2-, 3-, 5-, 6-, or
7-2H-furo[3,2-b]-pyranyl, 2-, 3-, 4-, 5-, 7-, or
8-5H-pyrido[2,3-d]-o-oxazinyl, 1-, 3-, or
5-1H-pyrazolo[4,3-d]-oxazolyl, 2-, 4-, or 54H-imidazo[4,5-d]
thiazolyl, 3-, 5-, or 8-pyrazino[2,3-d]pyridazinyl, 2-, 3-, 5-, or
6-imidazoyl-[2,1-b] thiazolyl, 1-, 3-, 6-, 7-, 8-, or
9-furo[3,4-c]cinnolinyl, 1-, 2-, 3-, 4-, 5-, 6-, 8-, 9-, 10, or 1
l-4H-pyrido[2,3-c]carbazolyl, 2-, 3-, 6-, or
7-imidazo[1,2-b][1,2,4]triazinyl, 7-benzo[b]thienyl, 2-, 4-, 5-,
6-, or 7-benzoxazolyl, 2-, 4-, 5-, 6-, or 7-benzimidazolyl, 2-, 4-,
4-, 5-, 6-, or 7-benzothiazolyl, 1-, 2-, 4-, 5-, 6-, 7-, 8-, or
9-benzoxapinyl, 2-, 4-, 5-, 6-, 7-, or 8-benzoxazinyl, 1-, 2-, 3-,
5-, 6-, 7-, 8-, 9-, 10-, or 11-1H-pyrrolo[1,2-b][2]benzazapinyl.
Typical fused heteroaryl groups include, but are not limited to 2-,
3-, 4-, 5-, 6-, 7-, or 8-quinolinyl, 1-, 3-, 4-, 5-, 6-, 7-, or
8-isoquinolinyl, 2-, 3-, 4-, 5-, 6-, or 7-indolyl, 2-, 3-, 4-, 5-,
6-, or 7-benzo[b]thienyl, 2-, 4-, 5-, 6-, or 7-benzoxazolyl, 2-,
4-, 5-, 6-, or 7-benzimidazolyl, 2-, 4-, 5-, 6-, or
7-benzothiazolyl.
[0083] As used herein, the term "heterocyclyl" or "heterocyclo"
refers to an optionally substituted, fully saturated or
unsaturated, aromatic or nonaromatic cyclic group, e.g., which is a
4- to 7-membered monocyclic, 7- to 12-membered bicyclic or 10- to
15-membered tricyclic ring system, which has at least one
heteroatom in at least one carbon atom-containing ring. Each ring
of the heterocyclic group containing a heteroatom may have 1, 2 or
3 heteroatoms selected from nitrogen atoms, oxygen atoms and sulfur
atoms, where the nitrogen and sulfur heteroatoms may also
optionally be oxidized. The heterocyclic group may be attached at a
heteroatom or a carbon atom.
[0084] Exemplary monocyclic heterocyclic groups include
pyrrolidinyl, pyrrolyl, pyrazolyl, oxetanyl, pyrazolinyl,
imidazolyl, imidazolinyl, imidazolidinyl, triazolyl, oxazolyl,
oxazolidinyl, isoxazolinyl, isoxazolyl, thiazolyl, thiadiazolyl,
thiazolidinyl, isothiazolyl, isothiazolidinyl, furyl,
tetrahydrofuryl, thienyl, oxadiazolyl, piperidinyl, piperazinyl,
2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolodinyl,
2-oxoazepinyl, azepinyl, 4-piperidonyl, pyridyl, pyrazinyl,
pyrimidinyl, pyridazinyl, tetrahydropyranyl, morpholinyl,
thiamorpholinyl, thiamorpholinyl sulfoxide, thiamorpholinyl
sulfone, 1,3-dioxolane and tetrahydro-1,1-dioxothienyl,
1,1,4-trioxo-1,2,5-thiadiazolidin-2-yl and the like.
[0085] Exemplary bicyclic heterocyclic groups include indolyl,
dihydroidolyl, benzothiazolyl, benzoxazinyl, benzoxazolyl,
benzothienyl, benzothiazinyl, quinuclidinyl, quinolinyl,
tetrahydroquinolinyl, decahydroquinolinyl, isoquinolinyl,
tetrahydroisoquinolinyl, decahydroisoquinolinyl, benzimidazolyl,
benzopyranyl, indolizinyl, benzofuryl, chromonyl, coumarinyl,
benzopyranyl, cinnolinyl, quinoxalinyl, indazolyl, pyrrolopyridyl,
furopyridinyl (such as furo[2,3-c]pyridinyl, furo[3,2-b]-pyridinyl]
or furo[2,3-b]pyridinyl), dihydroisoindolyl,
1,3-dioxo-1,3-dihydroisoindol-2-yl, dihydroquinazolinyl (such as
3,4-dihydro-4-oxo-quinazolinyl), phthalazinyl and the like.
[0086] Exemplary tricyclic heterocyclic groups include carbazolyl,
dibenzoazepinyl, dithienoazepinyl, benzindolyl, phenanthrolinyl,
acridinyl, phenanthridinyl, phenoxazinyl, phenothiazinyl,
xanthenyl, carbolinyl and the like.
[0087] The term "heterocyclyl" further refers to heterocyclic
groups as defined herein substituted with 1, 2 or 3 substituents
selected from the groups consisting of the following:
[0088] (a) alkyl;
[0089] (b) hydroxy (or protected hydroxy);
[0090] (c) halo;
[0091] (d) oxo, i.e., .dbd.O;
[0092] (e) amino, alkylamino or dialkylamino;
[0093] (f) alkoxy;
[0094] (g) cycloalkyl;
[0095] (h) carboxy;
[0096] (i) heterocyclooxy, wherein heterocyclooxy denotes a
heterocyclic group bonded through an oxygen bridge;
[0097] (j) alkyl-O--C(O)--;
[0098] (k) mercapto;
[0099] (l) nitro;
[0100] (m) cyano;
[0101] (n) sulfamoyl or sulfonamido;
[0102] (o) aryl;
[0103] (p) alkyl-C(O)--O--;
[0104] (q) aryl-C(O)--O--;
[0105] (r) aryl-S--;
[0106] (s) aryloxy;
[0107] (t) alkyl-S--;
[0108] (u) formyl, i.e., HC(O)--;
[0109] (v) carbamoyl;
[0110] (w) aryl-alkyl-; and
[0111] (x) aryl substituted with alkyl, cycloalkyl, alkoxy,
hydroxy, amino, alkyl-C(O)--NH--, alkylamino, dialkylamino or
halogen.
[0112] As used herein, the term "alkenyl" refers to a straight or
branched hydrocarbon group having 2 to 20 carbon atoms and that
contains at least one double bonds. The alkenyl groups preferably
have about 2 to 8 carbon atoms.
[0113] The term "aryl" means, unless otherwise stated, a
polyunsaturated, aromatic, hydrocarbon substituent, which can be a
single ring or multiple rings (preferably from 1 to 3 rings), which
are fused together or linked covalently. The term "heteroaryl"
refers to aryl groups (or rings) that contain from one to four
heteroatoms selected from N, O, and S, wherein the nitrogen and
sulfur atoms are optionally-oxidized, and the nitrogen atom(s) are
optionally quaternized. A heteroaryl group can be attached to the
remainder of the molecule through a heteroatom. Non-limiting
examples of aryl and heteroaryl groups include phenyl, 1-naphthyl,
2-naphthyl, 4-biphenyl, 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl,
3-pyrazolyl, 2-imidazolyl, 4-imidazolyl, pyrazinyl, 2-oxazolyl,
4-oxazolyl, 2-phenyl-4-oxazolyl, 5-oxazolyl, 3-isoxazolyl,
4-isoxazolyl, 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl,
2-furyl, 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl,
4-pyridyl, 2-pyrimidyl, 4-pyrimidyl, 5-benzothiazolyl, purinyl,
2-benzimidazolyl, 5-indolyl, 1-isoquinolyl, 5-isoquinolyl,
2-quinoxalinyl, 5-quinoxalinyl, 3-quinolyl, and 6-quinolyl.
Substituents for each of the above noted aryl and heteroaryl ring
systems are selected from the group of acceptable substituents
described below.
[0114] For brevity, the term "aryl" when used in combination with
other terms (e.g., aryloxy, arylthioxy, arylalkyl) includes both
aryl and heteroaryl rings as defined above. Thus, the term
"arylalkyl" is meant to include those radicals in which an aryl
group is attached to an alkyl group (e.g., benzyl, phenethyl,
pyridylmethyl and the like) including diose alkyl groups in which a
carbon atom (e.g., a methylene group) has been replaced by, for
example, an oxygen atom (e.g., phenoxymethyl, 2-pyridyloxymethyl,
3-(1-naphthyloxy)propyl, and the like).
[0115] Each of the above terms (e.g., "alkyl," "heteroalkyl."
"aryl" and "heteroaryl") include both substituted and unsubstituted
forms of the indicated radical. Preferred substituents for each
type of radical are provided below.
[0116] Substituents for the alkyl, and heteroalkyl radicals
(including those groups often referred to as alkylene, alkenyl,
heteroalkylene, heteroalkenyl, alkynyl, cycloalkyl,
heterocycloalkyl, cycloalkenyl, and heterocycloalkenyl) are
generally referred to as "alkyl substituents" and "heteroakyl
substituents," respectively, and they can be one or more of a
variety of groups selected from, but not limited to: --OR', .dbd.O,
.dbd.NR', .dbd.N--OR', --NR'R'', --SR', -halogen, --SiR'R'R''',
--OC(O)R', --C(O)R', --CO.sub.2R', --CONR'R'', --OC(O)NR'R'',
--NR''C(O)R', --NR'--C(O)NR''R''', --NR''C(O).sub.2R',
--NR--C(NR'R''R''').dbd.NR'''', --NR--C(NR'R'').dbd.NR''',
--S(O)R', --S(O).sub.2R', --S(O).sub.2NR'R'', --NRSO.sub.2R', --CN
and --NO.sub.2 in a number ranging from zero to (2m'+1), where m'
is the total number of carbon atoms in such radical. R', R'', R'''
and R'''' each preferably independently refer to hydrogen,
substituted or unsubstituted heteroalkyl, substituted or
unsubstituted aryl, e.g., aryl substituted with 1-3 halogens,
substituted or unsubstituted alkyl, alkoxy or thioalkoxy groups, or
arylalkyl groups. When a compound of the invention includes more
than one R group, for example, each of the R groups is
independently selected as are each R', R'', R''' and R'''' groups
when more than one of these groups is present. When R' and R'' are
attached to the same nitrogen atom, they can be combined with the
nitrogen atom to form a 5-, 6-, or 7-membered ring. For example,
--NR'R'' is meant to include, but not be limited to, 1-pyrrolidinyl
and 4-morpholinyl. From the above discussion of substituents, one
of skill in the art will understand that the term "alkyl" is meant
to include groups including carbon atoms bound to groups other than
hydrogen groups, such as haloalkyl (e.g., --CF.sub.3 and
--CH.sub.2CF.sub.3) and acyl (e.g., --C(O)CH.sub.3, --C(O)CF.sub.3,
--C(O)CH.sub.2OCH.sub.3, and the like).
[0117] Similar to the substituents described for the alkyl radical,
the aryl substituents and heteroaryl substituents are generally
referred to as "aryl substituents" and "heteroaryl substituents,"
respectively and are varied and selected from, for example:
halogen, --OR', .dbd.O, .dbd.NR', .dbd.N--OR', --NR'R'', --SR',
-halogen, --SiR'R''R''', --OC(O)R'', --C(O)R', --CO.sub.2R',
--CONR'R'', --OC(O)NR'R'', --NR''C(O)R', --NR*--C(O)NR''R''',
--NR''C(O).sub.2R', --NR--C(NR'R'').dbd.NR''', --S(O)R',
--S(O).sub.2R', --S(O).sub.2NR'R'', --NRSO.sub.2R', --CN and
--NO.sub.2, --R', --N.sub.3, --CH(Ph).sub.2,
fluoro(C.sub.1-C.sub.4)alkoxy, and fluoro(C.sub.1-C.sub.4)alkyl, in
a number ranging from zero to the total number of open valences on
the aromatic ring system; and where R', R'', R''' and R'''' are
preferably independently selected from hydrogen,
(C.sub.1-C.sub.8)alkyl and heteroalkyl, unsubstituted aryl and
heteroaryl, (unsubstituted aryl)-(C.sub.1-C.sub.4)alkyl, and
(unsubstituted aryl)oxy-(C.sub.1-C.sub.4)alkyl. When a compound of
the invention includes more than one R group, for example, each of
the R groups is independently selected as are each R', R'R'' and
R'''' groups when more than one of these groups is present.
[0118] Two of the aryl substituents on adjacent atoms of the aryl
or heteroaryl ring may optionally be replaced with a substituent of
the formula -T-C(O)--(CRR').sub.q-U-, wherein T and U are
independently --NR--, --O--, --CRR'-- or a single bond, and q is an
integer of from 0 to 3. Alternatively, two of the substituents on
adjacent atoms of the aryl or heteroaryl ring may optionally be
replaced with a substituent of the formula -A-(CH.sub.2).sub.r-B-,
wherein A and B are independently --CRR'--, --O--, --NR--, --S--,
--S(O)--, --S(O).sub.2--, --S(O).sub.2NR'-- or a single bond, and r
is an integer of from 1 to 4. One of the single bonds of the new
ring so formed may optionally be replaced with a double bond.
Alternatively, two of the substituents on adjacent atoms of the
aryl or heteroaryl ring may optionally be replaced with a
substituent of the formula --(CRR').sub.s--X--(CR''R''').sub.d,
where s and d are independently integers of from 0 to 3, and X is
--O--, --NR'--, --S--, --S(O)--, --S(O).sub.2--, or
--S(O).sub.2NR'--. The substituents R, R', R'' and R''' are
preferably independently selected from hydrogen or substituted or
unsubstituted (C.sub.1-C.sub.6) akyl.
[0119] As used herein, the term "heteroatom" includes oxygen (O),
nitrogen (N), sulfur (S), phosphorus (P) and silicon (Si).
[0120] As used herein, the term "aryloxy" refers to both an
--O-aryl and an --O-heteroaryl group, wherein aryl and heteroaryl
are defined herein.
[0121] As used herein, the term "pharmaceutically acceptable salts"
refers to salts that retain the biological effectiveness and
properties of the compounds of this invention and, which are not
biologically or otherwise undesirable. In many cases, the compounds
of the present invention are capable of forming acid and/or base
salts by virtue of the presence of amino and/or carboxyl groups or
groups similar thereto (e.g., phenol or hydroxyamic acid).
Pharmaceutically acceptable acid addition salts can be formed with
inorganic acids and organic acids. Inorganic acids from which salts
can be derived include, for example, hydrochloric acid, hydrobromic
acid, sulfuric acid, nitric acid, phosphoric acid, and the like.
Organic acids from which salts can be derived include, for example,
acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic
acid, maleic acid, malonic acid, succinic acid, fumaric acid,
tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic
acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic
acid, salicylic acid, and the like. Pharmaceutically acceptable
base addition salts can be formed with inorganic and organic bases.
Inorganic bases from which salts can be derived include, for
example, sodium, potassium, lithium, ammonium, calcium, magnesium,
iron, zinc, copper, manganese, aluminum, and the like; particularly
preferred are the ammonium, potassium, sodium, calcium and
magnesium salts.
[0122] Organic bases from which salts can be derived include, for
example, primary, secondary, and tertiary amines, substituted
amines including naturally occurring substituted amines, cyclic
amines, basic ion exchange resins, and the like, specifically such
as isopropylamine, trimethylamine, diethylamine, triethylamine,
tripropylamine, and ethanolamine. The pharmaceutically acceptable
salts of the present invention can be synthesized from a parent
compound, a basic or acidic moiety, by conventional chemical
methods. Generally, such salts can be prepared by reacting free
acid forms of these compounds with a stoichiometric amount of the
appropriate base (such as Na, Ca, Mg, or K hydroxide, carbonate,
bicarbonate, or the like), or by reacting free base forms of these
compounds with a stoichiometric amount of the appropriate acid.
Such reactions are typically carried out in water or in an organic
solvent, or in a mixture of the two. Generally, non-aqueous media
like ether, ethyl acetate, ethanol, isopropanol, or acetonitrile
are preferred, where practicable. Lists of additional suitable
salts can be found, e.g., in Remington's Pharmaceutical Sciences,
20th ed., Mack Publishing Company, Easton, Pa., (1985), which is
herein incorporated by reference.
[0123] As used herein, the term "pharmaceutically acceptable
carrier/excipient" includes any and all solvents, dispersion media,
coatings, surfactants, antioxidants, preservatives (e.g.,
antibacterial agents, antifungal agents), isotonic agents,
absorption delaying agents, salts, drugs, drug stabilizers,
binders, excipients, disintegration agents, lubricants, sweetening
agents, flavoring agents, dyes, such like materials and
combinations thereof, as would be known to one of ordinary skill in
the art (see, for example, Remington's Pharmaceutical Sciences,
18th Ed. Mack Printing Company, 1990, pp. 1289-1329, incorporated
herein by reference). Except in so far as any conventional carrier
is incompatible with the active ingredient, its use in the
therapeutic or pharmaceutical compositions is contemplated.
[0124] As used herein, the term "subject" refers to an animal.
Preferably, the animal is a mammal. A subject also refers to for
example, primates (e.g., humans), cows, sheep, goats, horses, dogs,
cats, rabbits, rats, mice, fish, birds and the like. In a preferred
embodiment, the subject is a human.
[0125] As used herein, the term "therapeutic combination" or
"combination" refers to a combination of one or more active drug
substances, i.e., compounds having a therapeutic utility.
Typically, each such compound in the therapeutic combinations of
the present invention will be present in a pharmaceutical
composition comprising that compound and a pharmaceutically
acceptable carrier. The compounds in a therapeutic combination of
the present invention may be administered simultaneously or
separately, as part of a regimen.
II. Compositions
[0126] In general, the present invention provides therapeutic
combinations, pharmaceutical compostions, and methods for treating
cancers using combination therapy. More specifically, the
combination of immunotherapy (such as using Toll-like Receptor
Ligand "TLRL" to activate DCs in innate immunity and link to
adaptive immunity) and targeted therapy (such as a PD-L/PD-1 Axis
antagonist) are used to treats cancers such as gastric cancer and
lung cancers.
[0127] In one aspect, the present invention provides therapeutic
combinations, or pharmaceutical compositions, comprising: (i) an
effective amount of a an effective amount of a PD-L/PD-1 Axis
antagonist; (ii) an effective amount of an immunotherapeutic that
is capable of activating a human dendritic cell, NK cell. Monocyte,
Macrophage or tumor cell, or a combination thereof; and optionally
(iii) one or more pharmaceutically acceptable carriers.
[0128] A therapeutic combination may be provided in a single
pharmaceutical composition so that both the targeted therapeutics
and the immunotherapeutic can be administered together. In
alternative embodiments, a therapeutic combination may be provided
using more than one pharmaceutical composition. In such
embodiments, a targeted therapeutic may be provided in one
pharmaceutical composition and an immunotherapeutic may be provided
in a second pharmaceutical composition so that the two compounds
can be administered separately such as, for example, at different
times, by different routes of administration, and the like. Thus,
it also may be possible to provide the targeted therapeutic and the
immunotherapeutic in different dosing regimens.
[0129] Unless otherwise indicated, reference to a compound can
include the compound in any pharmaceutically acceptable form,
including any isomer (e.g., diastereomer or enantiomer), salt,
solvate, polymorph, and the like. In particular, if a compound is
optically active, reference to the compound can include each of the
compound's enantiomers as well as racemic mixtures of the
enantiomers.
[0130] In general, the targeted therapeutics and the
immunotherapeutics are not linked to each other, such as by a
covalent linker.
A. PD-L/PD-1 Axis Antagonists
[0131] In general, the combination provided herein comprises an
entity, such as a PD-L/PD-1 Axis antagonist that is capable of
specifically binding to a particular target, such as PD-L1, PD-L2
or PD-1. The entity is capable of binding to PD-L1, PD-L2, or PD-1
specifically or preferably in comparison to a non-target.
[0132] By "specifically binds" or "preferably binds" herein is
meant that the binding between two binding partners (e.g., between
a targeting moiety and its binding partner) is selective for the
two binding partners and can be discriminated from unwanted or
non-specific interactions. For example, the ability of an
antigen-binding moiety to bind to a specific antigenic determinant
can be measured either through an enzyme-linked immunosorbent assay
(ELISA) or other techniques familiar to one of skill in the art,
e.g. surface plasmon resonance technique (analyzed on a BIAcore
instrument) (Liljeblad et al., Glyco J 17, 323-329 (2000)), and
traditional binding assays (Heeley, Endocr Res 28, 217-229 (2002)).
The terms "anti-[antigen] antibody" and "an antibody that binds to
[antigen]" refer to an antibody that is capable of binding the
respective antigen with sufficient affinity such that the antibody
is useful as a diagnostic and/or therapeutic agent in targeting the
antigen. In some embodiments, the extent of binding of an
anti-[antigen] antibody to an unrelated protein is less than about
10% of the binding of the antibody to the antigen as measured,
e.g., by a radioimmunoassay (RIA). In some embodiments, an antibody
that binds to [antigen] has a dissociation constant (KD) of <1
.mu.M, <100 nM, <10 nM, <1 nM, <0.1 nM, <0.01 nM, or
<0.001 nM (e.g. 10.sup.-8 M or less, e.g. from 10.sup.-8 M to
10.sup.-13 M, e.g., from 10.sup.-9 M to 10.sup.-13 M). It is
understood that the above definition is also applicable to
antigen-binding moieties that bind to an antigen.
[0133] By "PD-L/PD-1 Axis antagonist" herein is meant is a molecule
that inhibits the interaction of a PD-L/PD-1 axis binding partner
with either one or more of its binding partner, so as to remove
T-cell dysfunction resulting from signaling on the PD-L/PD-1
signaling axis--with a result being to restore or enhance T-cell
function {e.g., proliferation, cytokine production, target cell
killing). As used herein, a PD-L/PD-1 Axis antagonist includes a
PD-1 binding antagonist, a PD-L1 binding antagonist and a PD-L2
binding antagonist.
[0134] By "PD-1 binding antagonists" herein is meant is a molecule
that decreases, blocks, inhibits, abrogates or interferes with
signal transduction resulting from the interaction of PD-1 with one
or more of its binding partners, such as PD-L1, PD-L2. In some
embodiments, the PD-1 binding antagonist is a molecule that
inhibits the binding of PD-1 to its binding partners. In a specific
aspect, the PD-1 binding antagonist inhibits the binding of PD-1 to
PD-L1 and/or PD-L2. For example, PD-1 binding antagonists include
anti-PD-1 antibodies, antigen binding fragments thereof,
immunoadhesins, fusion proteins, oligopeptides and other molecules
that decrease, block, inhibit, abrogate or interfere with signal
transduction resulting from the interaction of PD-1 with PD-L1
and/or PD-L2. In one embodiment, a PD-1 binding antagonist reduces
the negative co-stimulatory signal mediated by or through cell
surface proteins expressed on T lymphocytes mediated signaling
through PD-1 so as render a dysfunctional T-cell less dysfunctional
(e.g., enhancing effector responses to antigen recognition). In
some embodiments, the PD-1 binding antagonist is an anti-PD-1
antibody. In a specific aspect, a PD-1 binding antagonist is MDX-1
106 described herein. In another specific aspect, a PD-1 binding
antagonist is Merck 3745 described herein. In another specific
aspect, a PD-1 binding antagonist is CT-01 1 described herein.
[0135] By "PD-L1 binding antagonists" herein is meant a molecule
that decreases, blocks, inhibits, abrogates or interferes with
signal transduction resulting from the interaction of PD-L1 with
either one or more of its binding partners, such as PD-1, B7-1. In
some embodiments, a PD-L1 binding antagonist is a molecule that
inhibits the binding of PD-L1 to its binding partners. In a
specific aspect, the PD-L1 binding antagonist inhibits binding of
PD-L1 to PD-1 and/or B7-1. In some embodiments, the PD-L1 binding
antagonists include anti-PD-L1 antibodies, antigen binding
fragments thereof, immunoadhesins, fusion proteins, oligopeptides
and other molecules that decrease, block, inhibit, abrogate or
interfere with signal transduction resulting from the interaction
of PD-L1 with one or more of its binding partners, such as PD-1,
B7-1. In one embodiment, a PD-L1 binding antagonist reduces the
negative co-stimulatory signal mediated by or through cell surface
proteins expressed on T lymphocytes mediated signaling through
PD-L1 so as to render a dysfunctional T-cell less dysfunctional
(e.g., enhancing effector responses to antigen recognition). In
some embodiments, a PD-L1 binding antagonist is an anti-PD-L1
antibody. In a specific aspect, an anti-PD-L1 antibody is
YW243.55.S70 described herein. In another specific aspect, an
anti-PD-L1 antibody is MDX-1105 described herein. In still another
specific aspect, an anti-PD-L1 antibody is MPDL3280A described
herein.
[0136] By "PD-L2 binding antagonists" herein is meant a molecule
that decreases, blocks, inhibits, abrogates or interferes with
signal transduction resulting from the interaction of PD-L2 with
either one or more of its binding partners, such as PD-1. In some
embodiments, a PD-L2 binding antagonist is a molecule that inhibits
the binding of PD-L2 to its binding partners. In a specific aspect,
the PD-L2 binding antagonist inhibits binding of PD-L2 to PD-1. In
some embodiments, the PD-L2 antagonists include anti-PD-L2
antibodies, antigen binding fragments thereof, immunoadhesins,
fusion proteins, oligopeptides and other molecules that decrease,
block, inhibit, abrogate or interfere with signal transduction
resulting from the interaction of PD-L2 with either one or more of
its binding partners, such as PD-1. In one embodiment, a PD-L2
binding antagonist reduces the negative co-stimulatory signal
mediated by or through cell surface proteins expressed on T
lymphocytes mediated signaling through PD-L2 so as render a
dysfunctional T-cell less dysfunctional (e.g., enhancing effector
responses to antigen recognition). In some embodiments, a PD-L2
binding antagonist is an immunoadhesin.
Antibodies
[0137] In some embodiments, the targeted therapeutic comprises an
antibody, or a functional fragment thereof.
[0138] By immunoglobulin" or "antibody" herein is meant a
full-length (i.e., naturally occurring or formed by normal
immunoglobulin gene fragment recombinatorial processes)
immunoglobulin molecule (e.g., an IgG antibody) or an
immunologically active (i.e., specifically binding) portion of an
immunoglobulin molecule, like an antibody fragment. An antibody or
antibody fragment may be conjugated or otherwise derivatized within
the scope of the claimed subject matter. Such antibodies include
IgG1, IgG2a, IgG3, IgG4 (and IgG4 subforms), as well as IgA
isotypes.
[0139] The term "antibody" herein is used in the broadest sense and
encompasses various antibody structures, including but not limited
to monoclonal antibodies, polyclonal antibodies, multispecific
antibodies (e.g. bispecific antibodies), and antibody fragments so
long as they exhibit the desired antigen-binding activity and
comprise an Fc region or a region equivalent to the Fc region of an
immunoglobulin The terms "full-length antibody", "intact antibody",
"and "whole antibody" are used herein interchangeably to refer to
an antibody having a structure substantially similar to a native
antibody structure or having heavy chains that contain an Fc region
as defined herein.
[0140] By "native antibodies" herein is meant naturally occurring
immunoglobulin molecules with varying structures. For example,
native IgG antibodies are heterotetrameric glycoproteins of about
150,000 daltons, composed of two identical light chains and two
identical heavy chains that are disulfide-bonded. From N- to
C-terminus, each heavy chain has a variable region (VH), also
called a variable heavy domain or a heavy chain variable domain,
followed by three constant domains (CHI, CH2, and CH3), also called
a heavy chain constant region. Similarly, from N- to C-terminus,
each light chain has a variable region (VL), also called a variable
light domain or a light chain variable domain, followed by a
constant light (CL) domain, also called a light chain constant
region. The light chain of an antibody may be assigned to one of
two types, called kappa (.kappa.) and lambda (.lamda.), based on
the amino acid sequence of its constant domain.
[0141] By "antibody fragment" herein is meant a molecule other than
an intact antibody that comprises a portion of an intact antibody
that binds the antigen to which the intact antibody binds. Examples
of antibody fragments include but are not limited to Fv, Fab. Fab',
Fab'-SH, F(ab')2, diabodies, linear antibodies, single-chain
antibody molecules (e.g. scFv), single-domain antibodies, and
multispecific antibodies formed from antibody fragments. For a
review of certain antibody fragments, see Hudson et al., Nat Med 9,
129-134 (2003). For a review of scFv fragments, see e.g.
Pliickthun, in The Pharmacology of Monoclonal Antibodies, vol. 113.
Rosenburg and Moore eds., Springer-Verlag, New York, pp. 269-315
(1994); see also WO 93/16185; and U.S. Pat. Nos. 5,571,894 and
5,587,458. For discussion of Fab and F(ab')2 fragments comprising
salvage receptor binding epitope residues and having increased in
vivo half-life, see U.S. Pat. No. 5,869,046. Diabodies are antibody
fragments with two antigen-binding sites that may be bivalent or
bispecific. See, for example, EP 404,097; WO 1993/01161; Hudson et
al., Nat Med 9, 129-134 (2003); and Hollinger et al., Proc Natl
Acad Sci USA 90, 6444-6448 (1993). Triabodies and tetrabodies are
also described in Hudson et al., Nat Med 9, 129-134 (2003).
Single-domain antibodies are antibody fragments comprising all or a
portion of the heavy chain variable domain or all or a portion of
the light chain variable domain of an antibody. In certain
embodiments, a single-domain antibody is a human single-domain
antibody (Domantis, Inc., Waltham, Mass.; see e.g. U.S. Pat. No.
6,248,516 B1). Antibody fragments can be made by various
techniques, including but not limited to proteolytic digestion of
an intact antibody as well as production by recombinant host cells
(e.g. E. coli or phage), as described herein.
[0142] By "antigen binding domain" herein is meant the part of an
antibody that comprises the area which specifically binds to and is
complementary to part or all of an antigen. An antigen binding
domain may be provided by, for example, one or more antibody
variable domains (also called antibody variable regions).
Particularly, an antigen binding domain comprises an antibody light
chain variable region (VL) and an antibody heavy chain variable
region (VH).
[0143] By "variable region" or "variable domain" herein is meant
the domain of an antibody heavy or light chain that is involved in
binding the antibody to antigen. The variable domains of the heavy
chain and light chain (VH and VL, respectively) of a native
antibody generally have similar structures, with each domain
comprising four conserved framework regions (FRs) and three
hypervariable regions (HVRs). See, e.g., Kindt et al., Kuby
Immunology, 6th ed., W.H. Freeman and Co., page 91 (2007). A single
VH or VL domain may be sufficient to confer antigen-binding
specificity.
[0144] By "hypervariable region" or "HVR" herein is meant each of
the regions of an antibody variable domain which are hypervariable
in sequence and/or form structurally defined loops ""hypervariable
loops"). Generally, native four-chain antibodies comprise six HVRs;
three in the VH (H1, H2, H3), and three in the VL (L1, L2, L3).
HVRs generally comprise amino acid residues from the hypervariable
loops and/or from the complementarity determining regions (CDRs),
the latter being of highest sequence variability and/or involved in
antigen recognition. With the exception of CDR1 in VH, CDRs
generally comprise the amino acid residues that form the
hypervariable loops. Hypervariable regions (HVRs) are also referred
to as "complementarity determining regions" (CDRs), and these terms
are used herein interchangeably in reference to portions of the
variable region that form the antigen binding regions. This
particular region has been described by Kabat et al., U.S. Dept. of
Health and Human Services, Sequences of Proteins of Immunological
Interest (1983) and by Chothia et al., J Mol Biol 196:901-917
(1987), where the definitions include overlapping or subsets of
amino acid residues when compared against each other. Nevertheless,
application of either definition to refer to a CDR of an antibody
or variants thereof is intended to be within the scope of the term
as defined and used herein. The exact residue numbers which
encompass a particular CDR will vary depending on the sequence and
size of the CDR. Those skilled in the art can routinely determine
which residues comprise a particular CDR given the variable region
amino acid sequence of the antibody.
[0145] The antibody of the present invention can be chimeric
antibodies, humanized antibodies, human antibodies, or antibody
fusion proteins.
[0146] By "chimeric antibody" herein is meant a recombinant protein
that contains the variable domains of both the heavy and light
antibody chains, including the complementarity determining regions
(CDRs) of an antibody derived from one species, preferably a rodent
antibody, more preferably a murine antibody, while the constant
domains of the antibody molecule are derived from those of a human
antibody. For veterinary applications, the constant domains of the
chimeric antibody may be derived from that of other species, such
as a subhuman primate, cat or dog.
[0147] By "humanized antibody" herein is meant a recombinant
protein in which the CDRs from an antibody from one species; e.g.,
a rodent antibody, are transferred from the heavy and light
variable chains of the rodent antibody into human heavy and light
variable domains. The constant domains of die antibody molecule are
derived from those of a human antibody. In some embodiments,
specific residues of the framework region of the humanized
antibody, particularly those that are touching or close to the CDR
sequences, may be modified, for example replaced with the
corresponding residues from the original rodent, subhuman primate,
or other antibody.
[0148] By "human antibody" herein is meant an antibody obtained,
for example, from transgenic mice that have been "engineered" to
produce specific human antibodies in response to antigenic
challenge. In this technique, elements of the human heavy and light
chain locus are introduced into strains of mice derived from
embryonic stem cell lines that contain targeted disruptions of the
endogenous heavy chain and light chain loci. The transgenic mice
can synthesize human antibodies specific for human antigens, and
the mice can be used to produce human antibody-secreting
hybridomas. Methods for obtaining human antibodies from transgenic
mice are described by Green et al. Nature Genet. 7: 13 (1994),
Lonberg et al, Nature 368:856 (1994), and Taylor et al, Int. Immun.
6:579 (1994). A fully human antibody also can be constructed by
genetic or chromosomal transfection methods, as well as phage
display technology, all of which are known in the art. See for
example, McCafferty et al, Nature 348:552-553 (1990) for the
production of human antibodies and fragments thereof in vitro, from
immunoglobulin variable domain gene repertoires from unimmunized
donors. In this technique, antibody variable domain genes are
cloned in-frame into either a major or minor coat protein gene of a
filamentous bacteriophage, and displayed as functional antibody
fragments on the surface of the phage particle. Because the
filamentous particle contains a single-stranded DNA copy of the
phage genome, selections based on the functional properties of the
antibody also result in selection of the gene encoding the antibody
exhibiting those properties. In this way, the phage mimics some of
the properties of the B cell. Phage display can be performed in a
variety of formats, for their review, see e.g. Johnson and
Chiswell, Current Opinion in Structural Biology 3:5564-571 (1993).
Human antibodies may also be generated by in vitro activated B
cells. See U.S. Pat. Nos. 5,567,610 and 5,229,275, which are
incorporated herein by reference in their entirety.
[0149] By "antibody fusion protein" herein is meant a
recombinantly-produced antigen-binding molecule in which two or
more of the same or different natural antibody, single-chain
antibody or antibody fragment segments with the same or different
specificities are linked. A fusion protein comprises at least one
specific binding site. Valency of the fusion protein indicates the
total number of binding arms or sites the fusion protein has to
antigen(s) or epitope(s); i.e., monovalent, bivalent, trivalent or
multivalent. The multivalency of the antibody fusion protein means
that it can take advantage of multiple interactions in binding to
an antigen, thus increasing the avidity of binding to the antigen,
or to different antigens. Specificity indicates how many different
types of antigen or epitope an antibody fusion protein is able to
bind; i.e., monospecific, bispecific, trispecific, multispecific.
Using these definitions, a natural antibody, e.g., an IgG, is
bivalent because it has two binding arms but is monospecific
because it binds to one type of antigen or epitope. A monospecific,
multivalent fusion protein has more than one binding site for the
same antigen or epitope. For example, a monospecific diabody is a
fusion protein with two binding sites reactive with the same
antigen. The fusion protein may comprise a multivalent or
multispecific combination of different antibody components or
multiple copies of the same antibody component. The fusion protein
may additionally comprise a therapeutic agent.
[0150] In some embodiments, the targeting moiety comprises a
probody, such as those disclosed in U.S. Pat. Nos. 8,518,404;
8,513,390; and US Pat. Appl. Pub. Nos.; 20120237977A1,
20120149061A1, 20130150558A1, the disclosures of which are
incorporated by reference in their entireties.
[0151] Probodies are monoclonal antibodies that are selectively
activated within the cancer microenvironment, focusing the activity
of therapeutic antibodies to tumors and sparing healthy tissue.
[0152] In general, the probody comprises at least an antibody- or
antibody fragment thereof (collectively referred to as "AB"),
capable of specifically binding a target, wherein the AB is
modified by a masking moiety (MM). When the AB is modified with a
MM and is in the presence of the target, specific binding of the AB
to its target is reduced or inhibited, as compared to the specific
binding of the AB not modified with an MM or the specific binding
of the parental AB to the target. The dissociation constant (Kd) of
the MM towards the AB is generally greater than the Kd of the AB
towards the target. When the AB is modified with a MM and is in the
presence of the target, specific binding of the AB to its target
can be reduced or inhibited, as compared to the specific binding of
the AB not modified with an MM or the specific binding of the
parental AB to the target. When an AB is coupled to or modified by
a MM, the MM can `mask` or reduce, or inhibit the specific binding
of the AB to its target. When an AB is coupled to or modified by a
MM, such coupling or modification can effect a structural change
which reduces or inhibits the ability of the AB to specifically
bind its target.
[0153] In some embodiments, the probody is an activatable
antibodies (AAs) where the AB modified by an MM can further include
one or more cleavable moieties (CM). Such AAs exhibit
activatable/switchable binding, to the AB's target. AAs generally
include an antibody or antibody fragment (AB), modified by or
coupled to a masking moiety (MM) and a modifiable or cleavable
moiety (CM). In some embodiments, the CM contains an amino acid
sequence that serves as a substrate for a protease of interest. In
other embodiments, the CM provides a cysteine-cysteine disulfide
bond that is cleavable by reduction. In yet other embodiments the
CM provides a photolytic substrate that is activatable by
photolysis.
[0154] The CM and AB of the AA may be selected so that the AB
represents a binding moiety for a target of interest, and the CM
represents a substrate for a protease that is co-localized with the
target at a treatment site in a subject. Alternatively, or in
addition, the CM is a cysteine-cysteine disulfide bond that is
cleavable as a result of reduction of this disulfide bond. AAs
contain at least one of a protease-cleavable CM or a
cysteine-cysteine disulfide bond, and in some embodiments, include
both kinds of CMs. The AAs can alternatively or further include a
photolabile substrate, activatable by a light source. The AAs
disclosed herein find particular use where, for example, a protease
capable of cleaving a site in the CM is present at relatively
higher levels in target-containing tissue of a treatment site (for
example diseased tissue; for example, for therapeutic treatment or
diagnostic treatment) than in tissue of non-treatment sites (for
example in healthy tissue). The AAs disclosed herein also find
particular use where, for example, a reducing agent capable of
reducing a site in the CM is present at relatively higher levels in
target-containing tissue of a treatment or diagnostic site than in
tissue of non-treatment non-diagnostic sites. The AAs disclosed
herein also find particular use where, for example, a light source,
for example, by way of laser, capable of photolysing a site in the
CM is introduced to a target-containing tissue of a treatment or
diagnostic site.
[0155] In some embodiments, AAs can provide for reduced toxicity
and/or adverse side effects that could otherwise result from
binding of the AB at non-treatment sites if the AB were not masked
or otherwise inhibited from binding its target. Where the AA
contains a CM that is cleavable by a reducing agent that
facilitates reduction of a disulfide bond, the ABs of such AAs may
be selected to exploit activation of an AB where a target of
interest is present at a desired treatment site characterized by
elevated levels of a reducing agent, such that the environment is
of a higher reduction potential than, for example, an environment
of a non-treatment site.
[0156] In general, an AA can be designed by selecting an AB of
interest and constructing the remainder of the AA so that, when
conformationally constrained, the MM provides for masking of the AB
or reduction of binding of the AB to its target. Structural design
criteria to be taken into account to provide for this functional
feature.
Anti-PD-1 Antibodies
[0157] In some embodiments, the TM is a monoclonal anti-PD-1
antibody.
[0158] Programmed Death-1 ("PD-1") is a receptor of PD-L1 (also
known as CD274, B7-H1, or B7-DC). PD-1 is an approximately 31 kD
type I membrane protein member of the extended CD28/CTLA4 family of
T cell regulators (Ishida, Y. et al. (1992) EMBO J. 11:3887-3895;
US Pat. Appl. Pub. No. 2007/0202100; 2008/0311117; 2009/00110667;
U.S. Pat. Nos. 6,808,710; 7,101,550; 7,488,802; 7,635,757;
7,722,868; PCT Publication No. WO 01/14557). In comparison to
CTLA4, PD-1 more broadly negatively regulates immune responses.
[0159] PD-1 is expressed on activated T cells, B cells, and
monocytes (Agata, Y. et al. (1996) Int. Immunol. 8(5):765-772;
Yamazaki, T. et al. (2002 J. Immunol. 169:5538-5545) and at low
levels in natural killer (NK) T cells (Nishimura, H. et al. (2000)
J. Exp. Med. 191:891-898; Martin-Orozco, N. et al. (2007), Semin.
Cancer Biol. 17(4):288-298).
[0160] The extracellular region of PD-1 consists of a single
immunoglobulin (Ig) V domain with 23% identity to the equivalent
domain in CTLA4 (Martin-Orozco, N. et al. (2007) Semin. Cancer
Biol. 17(4):288-298). The extracellular IgV domain is followed by a
transmembrane region and an intracellular tail. The intracellular
tail contains two phosphorylation sites located in an
immunoreceptor tyrosine-based inhibitory motif and an
immunoreceptor tyrosine-based switch motif, which suggests that
PD-1 negatively regulates TCR signals (Ishida, Y. et al. (1992 EMBO
J. 11:3887-3895; Blank, C. et al. (Epub 2006 Dec. 29) Immunol.
Immunother. 56(5):739-745).
[0161] Antibodies capable of immunospecifically binding to murine
PD-1 have been reported (see, e.g., Agata, T. et al. (1996) Int.
Immunol. 8(5):765-772).
[0162] Anti-PD-1 antibodies bind to PD-1 and enhance T-cell
function to upregulate cell-mediated immune responses and for the
treatment of T cell dysfunctional disorders, such as tumor
immunity.
[0163] In some embodiments, the anti-PD-1 antibody is MK-3475
(formerly Iambrolizumab, Merck), AMP-514, AMP-224
(MedImmune/AstraZeneca), BMS-936558 (MDX-1106, Bristol-Myers
Squibb), or CT-011 (Curetech).
[0164] Pembrolizumab (MK-3475) is a humanized, monoclonal anti-PD-1
antibody designed to reactivate anti-tumor immunity. Pembrolizumab
exerts dual ligand blockade of the PD-1 pathway by inhibiting the
interaction of PD-1 on T cells with its ligands PD-L1 and
PD-L2.
[0165] In some embodiments, the anti-PD-1 antibody is one of the
antibodies disclosed in U.S. Pat. No. 8,354,509, and U.S. Pat. No.
8,168,757, the disclosure of which is incorporated by reference in
their entirety.
[0166] Nivolumab (also known as BMS-936558 or MDX1106, is a fully
human IgG4 monoclonal antibody developed by Bristol-Myers Squibb
for the treatment of cancer.
[0167] In some embodiments, the anti-PD-1 antibody is one of the
antibodies disclosed in WO2004/056875, U.S. Pat. No. 7,488,802 and
U.S. Pat. No. 8,008,449, the disclosure of which is incorporated by
reference in their entirety.
[0168] AMP-514 and AMP-224 are an anti-programmed cell death 1
(PD-1) monoclonal antibody (mAb) developed by Amplimmune, which was
acquired by MedImmune.
[0169] In some embodiments, the anti-PD-1 antibody is one of the
antibodies disclosed in US Appl. Pub. No. 20140044738, the
disclosure of which is incorporated by reference in their
entirety.
[0170] In some embodiments, the six CDRs are: (A) the three light
chain and the three heavy chain CDRs of anti-PD-1 antibody 1E3; (B)
the three light chain and the three heavy chain CDRs of anti-PD-1
antibody 1E8; or (C) the three light chain and the three heavy
chain CDRs of anti-PD-1 antibody 1H3.
[0171] Pidilizumab (CT-011) is an anti-PD-1 monoclonal antibody
developed by Israel-based Curetech Ltd.
[0172] In some embodiments, the anti-PD-1 antibody is one of the
antibodies disclosed in US Pat. Appl. Pub. Nos. 20080025980 and
20130022595, the disclosure of which is incorporated by reference
in their entirety.
Anti-PD-L1 Antibodies
[0173] In some embodiments, the TM is a monoclonal anti-PD-L1
antibody.
[0174] Programmed cell death 1 ligand 1 (PD-L1, also known as CD274
and B7-H1) is a ligand for PD-1, found on activated T cells, B
cells, myeloid cells and macrophages. Although there are two
endogenous ligands for PD-1, PD-L1 and PD-L2, anti-tumor therapies
have focused on anti-PD-L1 antibodies. The complex of PD-1 and
PD-L1 inhibits proliferation of CD8+ T cells and reduces the immune
response (Topalian et al., 2012, N Engl J Med 366:2443-54; Brahmer
et al., 2012, N Eng J Med 366:2455-65). Anti-PD-L1 antibodies have
been used for treatment of non-small cell lung cancer, melanoma,
colorectal cancer, renal-cell cancer, pancreatic cancer, gastric
cancer, ovarian cancer, breast cancer, and hematologic malignancies
(Brahmer et al., N Eng J Med 366:2455-65; Ott et al., 2013, Clin
Cancer Res 19:5300-9; Radvanyi et al., 2013, Clin Cancer Res
19:5541; Menzies & Long, 2013, Ther Adv Med Oncol 5:278-85;
Berger et al., 2008, Clin Cancer Res 14:13044-51). PD-L1 is a B7
family member that is expressed on many cell types, including APCs
and activated T cells (Yamazaki et al. (2002) J. Immunol.
169:5538). PD-L1 binds to both PD-1 and B7-1. Both binding of
T-cell-expressed B7-1 by PD-L1 and binding of T-cell-expressed
PD-L1 by B7-1 result in T cell inhibition (Butte et al. (2007)
Immunity 27:111). There is also evidence that, like other B7 family
members, PD-L1 can also provide costimulatory signals to T cells
(Subudhi et al. (2004) J. Clin. Invest. 113:694; Tamura et al.
(2001) Blood 97:1809).
[0175] By "PD-L1" herein is meant to include any variants or
isoforms which are naturally expressed by cells, and/or fragments
thereof having at least one biological activity of the full-length
polypeptide, unless otherwise expressly defined. In addition, the
term "PD-L1" includes PD-L1 (Freeman et al. (2000) J. Exp. Med.
192:1027) and any variants or isoforms which are naturally
expressed by cells, and/or fragments thereof having at least one
biological activity of the full-length polypeptides. For example,
PD-L1 sequences from different species, including humans, are well
known in the art (see, for example, herein incorporated in their
entirety by reference, Chen et al., U.S. Pat. No. 6,803,192, which
discloses human and mouse PD-L1 sequences; Wood et al., U.S. Pat.
No. 7,105,328, which discloses human PD-L1 sequences.
[0176] Anti-PD-L1 antibodies bind to PD-L1 and enhance T-cell
function to upregulate cell-mediated immune responses and for the
treatment of T cell dysfunctional disorders, such as tumor
immunity.
[0177] In some embodiments, the anti-PD-L1 antibody is MPDL3280A
and YW243.55.S70, (Genentech/Roche), MEDI-4736
(MedImmune/AstraZeneca), BMS-936559 (MDX-1105, Bristol-Myers
Squibb), and MSB0010718C (EMD Serono/Merck KGaA).
[0178] MPDL3280A (Genentech) is an engineered anti-PD-L1 antibody
designed to target PD-L1 expressed on tumor cells and
tumor-infiltrating immune cells. MPDL3280A is designed to prevent
PD-L1 from binding to PD-1 and B7.1. This blockade of PD-L1 may
enable the activation of T cells, restoring their ability to detect
and attack tumor cells. MPDL3280A contains an engineered fragment
crystallizable (Fc) domain designed to optimize efficacy and safety
by minimizing antibody-dependent cellular cytotoxicity (ADCC).
[0179] In some embodiments, the anti-PD-L1 antibody is one of the
antibodies disclosed in U.S. Pat. No. 7,943,743, the disclosure of
which is incorporated by reference in their entirety.
[0180] BMS-936559 (MDX-1105, Bristol-Myers Squibb) is a fully human
IgG4 anti-PD-L1 mAb that inhibits the binding of the PD-L1 ligand
to both PD-1 and CD80.
[0181] In some embodiments, the anti-PD-L1 antibody is one of the
antibodies disclosed in U.S. Pat. No. 7,943,743, the disclosure of
which is incorporated by reference in their entirety.
[0182] MSB0010718C (EMD Serono of Merck KGaA) is fully human IgG1
monoclonal antibody that binds to PD-L1.
[0183] In some embodiments, the anti-PD-L1 antibody is one of the
antibodies disclosed in WO 2013079174 A1, the disclosure of which
is incorporated by reference in their entirety.
[0184] MEDI4736 (MedImmune/AstraZeneca) is a human IgG1 antibody
which binds specifically to PD-L1, preventing binding to PD-1 and
CD80.
[0185] In some embodiments, the anti-PD-L1 antibody is one of the
antibodies disclosed in WO 2011066389 A1 and U.S. Pat. No.
8,779,108, the disclosure of which is incorporated by reference in
their entirety.
[0186] In some embodiments, the anti-PD-L1 antibody is one of the
antibodies disclosed in U.S. Pat. No. 8,552,154, the disclosure of
which is incorporated by reference in their entirety.
[0187] In some embodiments, the targeting moiety comprises a Fab,
Fab', F(ab')2, single domain antibody, T and Abs dimer, Fv, scFv,
dsFv, ds-scFv, Fd, linear antibody, minibody, diabody, bispecific
antibody fragment, bibody, tribody, sc-diabody, kappa (lamda) body,
BiTE, DVD-Ig, SIP, SMIP, DART, or an antibody analogue comprising
one or more CDRs.
PD-L/PD-1 Axis Antagonist Comprising a Targeting Moiety
[0188] In some aspects, the PD-L/PD-1 Axis antagonist is a targeted
therapeutic comprise a targeting moiety, such as an ADC.
[0189] By "targeting moiety (TM)" or "targeting agent" here in is
meant a molecule, complex, or aggregate, that binds specifically or
selectively to a target molecule, cell, particle, tissue or
aggregate, which generally is referred to as a "target" or a
"marker," and these are discussed in further detail herein.
[0190] In some embodiments, the targeting moiety comprises an
immunoglobulin, a protein, a peptide, a small molecule, a
nanoparticle, or a nucleic acid.
[0191] Exemplary targeting agents such as antibodies (e.g.,
chimeric, humanized and human), ligands for receptors, lecitins,
and saccharides, and substrate for certain enzymes are recognized
in the art and are useful without limitation in practicing the
present invention. Other targeting agents include a class of
compounds that do not include specific molecular recognition motifs
include nanoparticles, macromolecules such as poly(ethylene
glycol), polysaccharide, and polyamino acids which add molecular
mass to the activating moiety. The additional molecular mass
affects the pharmacokinetics of the activating moiety, e.g., serum
half-life.
[0192] In some embodiments, a targeting moiety is an antibody,
antibody fragment, bispecific antibody or other antibody-based
molecule or compound. However, other examples of targeting moieties
are known in the art and may be used, such as aptamers, avimers,
receptor-binding ligands, nucleic acids, biotin-avidin binding
pairs, binding peptides or proteins, etc. The terms "targeting
moiety" and "binding moiety" are used synonymously herein.
[0193] By "target" or "marker" herein is meant any entity that is
capable of specifically binding to a particular targeting moiety.
In some embodiments, targets are specifically associated with one
or more particular cell or tissue types. In some embodiments,
targets are specifically associated with one or more particular
disease states. In some embodiments, targets are specifically
associated with one or more particular developmental stages. For
example, a cell type specific marker is typically expressed at
levels at least 2 fold greater in that cell type than in a
reference population of cells. In some embodiments, the cell type
specific marker is present at levels at least 3 fold, at least 4
fold, at least 5 fold, at least 6 fold, at least 7 fold, at least 8
fold, at least 9 fold, at least 10 fold, at least 50 fold, at least
100 fold, or at least 1,000 fold greater than its average
expression in a reference population. Detection or measurement of a
cell type specific marker may make it possible to distinguish the
cell type or types of interest from cells of many, most, or all
other types. In some embodiments, a target can comprise a protein,
a carbohydrate, a lipid, and/or a nucleic acid, as described
herein.
[0194] A substance is considered to be "targeted" for the purposes
described herein if it specifically binds to a nucleic acid
targeting moiety. In some embodiments, a nucleic acid targeting
moiety specifically binds to a target under stringent conditions.
An inventive complex or compound comprising targeting moiety is
considered to be "targeted" if the targeting moiety specifically
binds to a target, thereby delivering the entire complex or
compound composition to a specific organ, tissue, cell,
extracellular matrix component, and/or intracellular
compartment.
[0195] In certain embodiments, compound in accordance with the
present invention comprise a targeting moiety which specifically
binds to one or more targets (e.g. antigens) associated with an
organ, tissue, cell, extracellular matrix component, and/or
intracellular compartment. In some embodiments, compounds comprise
a targeting moiety which specifically binds to targets associated
with a particular organ or organ system. In some embodiments,
compounds in accordance with the present invention comprise a
nuclei targeting moiety which specifically binds to one or more
intracellular targets (e.g. organelle, intracellular protein). In
some embodiments, compounds comprise a targeting moiety which
specifically binds to targets associated with diseased organs,
tissues, cells, extracellular matrix components, and/or
intracellular compartments. In some embodiments, compounds comprise
a targeting moiety which specifically binds to targets associated
with particular cell types (e.g. endothelial cells, cancer cells,
malignant cells, prostate cancer cells, etc.).
[0196] In some embodiments, compounds in accordance with the
present invention comprise a targeting moiety which binds to a
target that is specific for one or more particular tissue types
(e.g. liver tissue vs. prostate tissue). In some embodiments,
compounds in accordance with the present invention comprise a
targeting moiety which binds to a target that is specific for one
or more particular cell types (e.g. T cells vs. B cells). In some
embodiments, compounds in accordance with the present invention
comprise a targeting moiety which binds to a target that is
specific for one or more particular disease states (e.g. tumor
cells vs. healthy cells). In some embodiments, compounds in
accordance with the present invention comprise a targeting moiety
which binds to a target that is specific for one or more particular
developmental stages (e.g. stem cells vs. differentiated
cells).
[0197] In some embodiments, a target may be a marker that is
exclusively or primarily associated with one or a few cell types,
with one or a few diseases, and/or with one or a few developmental
stages. A cell type specific marker is typically expressed at
levels at least 2 fold greater in that cell type than in a
reference population of cells which may consist, for example, of a
mixture containing cells from a plurality (e.g., 5-10 or more) of
different tissues or organs in approximately equal amounts. In some
embodiments, the cell type specific marker is present at levels at
least 3 fold, at least 4 fold, at least 5 fold, at least 6 fold, at
least 7 fold, at least 8 fold, at least 9 fold, at least 10 fold,
at least 50 fold, at least 100 fold, or at least 1000 fold greater
than its average expression in a reference population. Detection or
measurement of a cell type specific marker may make it possible to
distinguish the cell type or types of interest from cells of many,
most, or all other types.
[0198] In some embodiments, a target comprises a protein, a
carbohydrate, a lipid, and/or a nucleic acid. In some embodiments,
a target comprises a protein and/or characteristic portion thereof,
such as a tumor-marker, integrin, cell surface receptor,
transmembrane protein, intercellular protein, ion channel, membrane
transporter protein, enzyme, antibody, chimeric protein,
glycoprotein, etc. In some embodiments, a target comprises a
carbohydrate and/or characteristic portion thereof, such as a
glycoprotein, sugar (e.g., monosaccharide, disaccharide,
polysaccharide), glycocalyx (i.e., the carbohydrate-rich peripheral
zone on the outside surface of most eukaryotic cells) etc. In some
embodiments, a target comprises a lipid and/or characteristic
portion thereof, such as an oil, fatty acid, glyceride, hormone,
steroid (e.g., cholesterol, bile acid), vitamin (e.g. vitamin E),
phospholipid, sphingolipid, lipoprotein, etc. In some embodiments,
a target comprises a nucleic acid and/or characteristic portion
thereof, such as a DNA nucleic acid; RNA nucleic acid; modified DNA
nucleic acid; modified RNA nucleic acid; nucleic acid that includes
any combination of DNA, RNA, modified DNA, and modified RNA.
[0199] Numerous markers are known in the art. Typical markers
include cell surface proteins, e.g., receptors. Exemplary receptors
include, but are not limited to, the transferrin receptor; LDL
receptor; growth factor receptors such as epidermal growth factor
receptor family members (e.g., EGFR, Her2, Her3, Her4) or vascular
endothelial growth factor receptors, cytokine receptors, cell
adhesion molecules, integrins, selectins, and CD molecules. The
marker can be a molecule that is present exclusively or in higher
amounts on a malignant cell, e.g., a tumor antigen.
[0200] In some embodiments, the targeting moiety binds to a tumor
cell specifically or preferably in comparison to a non-tumor
cell.
[0201] The binding of target moiety to tumor cell can be measured
using assays known in the art.
[0202] In some embodiments, the tumor cell is of a carcinoma, a
sarcoma, a lymphoma, a myeloma, or a central nervous system
cancer.
[0203] In some embodiments, the targeting moiety is capable of
binding to a tumor antigen specifically or preferably in comparison
to a non-tumor antigen.
[0204] In certain specific embodiments, a target is a tumor marker.
In some embodiments, a tumor marker is an antigen that is present
in a tumor that is not present in normal organs, tissues, and/or
cells. In some embodiments, a tumor marker is an antigen that is
more prevalent in a tumor than in normal organs, tissues, and/or
cells. In some embodiments, a tumor marker is an antigen that is
more prevalent in malignant cancer cells than in normal cells.
[0205] In some embodiments, the targeting moiety comprises folic
acid or a derivative thereof.
[0206] In recent years, research on folic acid had made great
progress. Folic acid is a small molecule vitamin that is necessary
for cell division. Tumor cells divide abnormally and there is a
high expression of folate receptor (FR) on tumor cell surface to
capture enough folic acid to support cell division.
[0207] Data indicate FR expression in tumor cells is 20-200 times
higher than normal cells. The expression rate of FR in various
malignant tumors are: 82% in ovarian cancer, 66% in non-small cell
lung cancer, 64% in kidney cancer, 34% in colon cancer, and 29% in
breast cancer (Xia W, Low P S. Late-targeted therapies for cancer.
J Med Chem. 2010; 14; 53 (19):6811-24). The expression rate of FA
and the degree of malignancy of epithelial tumor invasion and
metastasis is positively correlated. FA enters cell through FR
mediated endocytosis, and FA through its carboxyl group forms FA
complexes with drugs which enter the cells. Under acidic conditions
(pH value of 5), FR separates from the FA, and FA releases drugs
into the cytoplasm.
[0208] Clinically, the system can be used to deliver drugs
selectively attack the tumor cells. Folic acid has small molecular
weight, has non-immunogenicity and high stability, and is
inexpensive to synthesis. More importantly, chemical coupling
between the drug and the carrier is simple, and as such using FA as
targeting molecule to construct drug delivery system has become a
research hotspot for cancer treatment. Currently EC 145 (FA
chemotherapy drug conjugate compound) that is in clinical trials
can effectively attack cancer cells (Pribble P and Edelman M J. EC
145: a novel targeted agent for adenocarcinoma of the lung. Expert
Opin. Investig. Drugs (2012) 21:755-761).
[0209] In some embodiments, the targeting moiety comprises
extracellular domains (ECD) or soluble form of PD-1, PDL-1, CTLA4,
CD47, BTLA, KIR, TIM3, 4-1BB, and LAG3, full length of partial of a
surface ligand Amphiregulin, Betacellulin, EGF, Ephrin, Epigen,
Epiregulin, IGF, Neuregulin, TGF, TRAIL, or VEGF.
[0210] In some embodiments, the targeting moiety comprises a Fab,
Fab', F(ab')2, single domain antibody, T and Abs dimer, Fv, scFv,
dsFv, ds-scFv, Fd, linear antibody, minibody, diabody, bispecific
antibody fragment, bibody, tribody, sc-diabody, kappa (lamda) body,
BiTE, DVD-Ig, SIP, SMIP, DART, or an antibody analogue comprising
one or more CDRs.
[0211] In some embodiments, the targeting moiety is an antibody, or
antibody fragment, that is selected based on its specificity for an
antigen expressed on a target cell, or at a target site, of
interest. A wide variety of tumor-specific or other
disease-specific antigens have been identified and antibodies to
those antigens have been used or proposed for use in the treatment
of such tumors or other diseases. The antibodies that are known in
the art can be used in the compounds of the invention, in
particular for the treatment of the disease with which the target
antigen is associated. Examples of target antigens (and their
associated diseases) to which an antibody-linker-drug conjugate of
the invention can be targeted include: CD2, CD19, CD20, CD22, CD27,
CD33, CD37, CD38, CD40, CD44, CD47, CD52, CD56, CD70, CD79, CD137,
4-1BB, 5T4, AGS-5, AGS-16, Angiopoietin 2, B7.1, B7.2, B7DC, B7H1,
B7H2, B7H3, BT-062, BTLA, CAIX, Carcinoembryonic antigen, CTLA4,
Cripto, ED-B, ErbB1, ErbB2, ErbB3, ErbB4, EGFL7, EpCAM, EphA2,
EphA3, EphB2, FAP, Fibronectin, Folate Receptor, Ganglioside GM3,
GD2, glucocorticoid-induced tumor necrosis factor receptor (GITR),
gp100, gpA33, GPNMB, ICOS, IGF1R, Integrin .alpha..nu., Integrin
.alpha..nu..beta., KIR, LAG-3, Lewis Y, Mesothelin, c-MET, MN
Carbonic anhydrase IX, MUC1, MUC16, Nectin-4, NKGD2, NOTCH, OX40,
OX40L, PD-1, PDL1, PSCA, PSMA, RANKL, ROR1, ROR2, SLC44A4,
Syndecan-1, TACI, TAG-72, Tenascin, TIM3, TRAILR1, TRAILR2,
VEGFR-1, VEGFR-2, VEGFR-3.
[0212] In some embodiments, the targeting moiety comprises a
particle (target particle), preferably a nanoparticle, optionally a
targeted nanoparticle that attached to a targeting molecule that
can binds specifically or preferably to a target. In some
embodiments, the targeting particle by itself guides die compound
of the present invention (such as by enrichment in tumor cells or
tissue) and there is no additional targeting molecules attached
therein.
[0213] By "nanoparticle" herein is meant any particle having a
diameter of less than 1000 nm. In some embodiments, a therapeutic
agent and/or targeting molecule can be associated with the
polymeric matrix. In some embodiments, the targeting molecule can
be covalently associated with the surface of a polymeric matrix. In
some embodiments, covalent association is mediated by a linker. In
some embodiments, the therapeutic agent can be associated with the
surface of, encapsulated within, surrounded by, and/or dispersed
throughout the polymeric matrix. U.S. Pat. No. 8,246,968, which is
incorporated in its entirety.
[0214] In general, nanoparticles of the present invention comprise
any type of particle. Any particle can be used in accordance with
the present invention. In some embodiments, particles are
biodegradable and biocompatible. In general, a biocompatible
substance is not toxic to cells. In some embodiments, a substance
is considered to be biocompatible if its addition to cells results
in less than a certain threshold of cell death. In some
embodiments, a substance is considered to be biocompatible if its
addition to cells does not induce adverse effects. In general, a
biodegradable substance is one that undergoes breakdown under
physiological conditions over the course of a therapeutically
relevant time period (e.g., weeks, months, or years). In some
embodiments, a biodegradable substance is a substance that can be
broken down by cellular machinery. In some embodiments, a
biodegradable substance is a substance that can be broken down by
chemical processes. In some embodiments, a particle is a substance
that is both biocompatible and biodegradable. In some embodiments,
a particle is a substance that is biocompatible, but not
biodegradable. In some embodiments, a particle is a substance that
is biodegradable, but not biocompatible.
[0215] In some embodiments, particles are greater in size than the
renal excretion limit (e.g. particles having diameters of greater
than 6 nm). In some embodiments, particles are small enough to
avoid clearance of particles from the bloodstream by the liver
(e.g. particles having diameters of less than 1000 nm). In general,
phytochemical features of particles should allow a targeted
particle to circulate longer in plasma by decreasing renal
excretion and liver clearance.
[0216] It is often desirable to use a population of particles that
is relatively uniform in terms of size, shape, and/or composition
so that each particle has similar properties. For example, at least
80%, at least 90%, or at least 95% of the particles may have a
diameter or greatest dimension that falls within 5%, 10%, or 20% of
the average diameter or greatest dimension. In some embodiments, a
population of particles may be heterogeneous with respect to size,
shape, and/or composition.
[0217] Zeta potential is a measurement of surface potential of a
particle. In some embodiments, particles have a zeta potential
ranging between -50 mV and +50 mV. In some embodiments, particles
have a zeta potential ranging between -25 mV and +25 mV. In some
embodiments, particles have a zeta potential ranging between -10 mV
and +10 mV. In some embodiments, particles have a zeta potential
ranging between -5 mV and +5 mV. In some embodiments, particles
have a zeta potential ranging between 0 mV and +50 mV. In some
embodiments, particles have a zeta potential ranging between 0 mV
and +25 mV.
[0218] In some embodiments, particles have a zeta potential ranging
between 0 mV and +10 mV. In some embodiments, particles have a zeta
potential ranging between 0 mV and +5 mV. In some embodiments,
particles have a zeta potential ranging between -50 mV and 0 mV. In
some embodiments, particles have a zeta potential ranging between
-25 mV and 0 mV. In some embodiments, particles have a zeta
potential ranging between -10 mV and 0 mV. In some embodiments,
particles have a zeta potential ranging between -5 mV and 0 mV. In
some embodiments, particles have a substantially neutral zeta
potential (i.e. approximately 0 mV).
[0219] A variety of different particles can be used in accordance
with the present invention. In some embodiments, particles are
spheres or spheroids. In some embodiments, particles are spheres or
spheroids. In some embodiments, particles are flat or plate-shaped.
In some embodiments, particles are cubes or cuboids. In some
embodiments, particles are ovals or ellipses. In some embodiments,
particles are cylinders, cones, or pyramids.
[0220] In some embodiments, particles are microparticles (e.g.
microspheres). In general, a "microparticle" refers to any particle
having a diameter of less than 1000 .mu.m. In some embodiments,
particles are picoparticles (e.g. picospheres). In general, a
"picoparticle" refers to any particle having a diameter of less
than 1 nm. In some embodiments, particles are liposomes. In some
embodiments, particles are micelles.
[0221] Particles can be solid or hollow and can comprise one or
more layers (e.g., nanoshells, nanorings). In some embodiments,
each layer has a unique composition and unique properties relative
to the other layers). For example, particles may have a core/shell
structure, wherein the core is one layer and the shell is a second
layer. Particles may comprise a plurality of different layers. In
some embodiments, one layer may be substantially cross-linked, a
second layer is not substantially cross-linked, and so forth. In
some embodiments, one, a few, or all of the different layers may
comprise one or more therapeutic or diagnostic agents to be
delivered. In some embodiments, one layer comprises an agent to be
delivered, a second layer does not comprise an agent to be
delivered, and so forth. In some embodiments, each individual layer
comprises a different agent or set of agents to be delivered.
[0222] In some embodiments, a particle is porous, by which is meant
that the particle contains holes or channels, which are typically
small compared with the size of a particle. For example, a particle
may be a porous silica particle, e.g., a mesoporous silica
nanoparticle or may have a coating of mesoporous silica (Lin et
al., 2005, J. Am. Chem. Soc., 17:4570). Particles may have pores
ranging from about 1 nm to about 50 nm in diameter, e.g., between
about 1 and 20 nm in diameter. Between about 10% and 95% of the
volume of a particle may consist of voids within the pores or
channels.
[0223] Particles may have a coating layer. Use of a biocompatible
coating layer can be advantageous, e.g., if the particles contain
materials that are toxic to cells. Suitable coating materials
include, but are not limited to, natural proteins such as bovine
serum albumin (BSA), biocompatible hydrophilic polymers such as
polyethylene glycol (PEG) or a PEG derivative, phospholipid-(PEG),
silica, lipids, polymers, carbohydrates such as dextran, other
nanoparticles that can be associated with inventive nanoparticles
etc.
[0224] Coatings may be applied or assembled in a variety of ways
such as by dipping, using a layer-by-layer technique, by
self-assembly, conjugation, etc. Self-assembly refers to a process
of spontaneous assembly of a higher order structure that relies on
the natural attraction of the components of the higher order
structure (e.g., molecules) for each other. It typically occurs
through random movements of the molecules and formation of bonds
based on size, shape, composition, or chemical properties.
[0225] Examples of polymers include polyalkylenes (e.g.
polyethylenes), polycarbonates (e.g. poly(1,3-dioxan-2one)),
polyanhydrides (e.g. poly(sebacic anhydride)), polyhydroxyacids
(e.g. poly(.beta.-hydroxyalkanoate)), polyfumarates,
polycaprolactones, polyamides (e.g. polycaprolactam), polyacetals,
polyethers, polyesters (e.g. polylactide, polyglycolide),
poly(orthoesters), polyvinyl alcohols, polyurethanes,
polyphosphazenes, polyacrylates, polymethacrylates,
polycyanoacrylates, polyureas, polystyrenes, and polyamines. In
some embodiments, polymers in accordance with the present invention
include polymers which have been approved for use in humans by the
U.S. Food and Drug Administration (FDA) under 21 C.F.R. .sctn.
177.2600, including but not limited to polyesters (e.g. polylactic
acid, polyglycolic acid, poly(lactic-co-glycolic acid),
polycaprolactone, polyvalerolactone, poly(1,3-dioxan-2one));
polyanhydrides (e.g. poly(sebacic anhydride)); polyethers (e.g.,
polyethylene glycol); polyurethanes; polymethacrylates;
polyacrylates; and polycyanoacrylates.
[0226] In some embodiments, particles can be non-polymeric
particles (e.g. metal particles, quantum dots, ceramic particles,
polymers comprising inorganic materials, bone-derived materials,
bone substitutes, viral particles, etc.). In some embodiments, a
therapeutic or diagnostic agent to be delivered can be associated
with the surface of such a non-polymeric particle. In some
embodiments, a non-polymeric particle is an aggregate of
non-polymeric components, such as an aggregate of metal atoms (e.g.
gold atoms). In some embodiments, a therapeutic or diagnostic agent
to be delivered can be associated with the surface of and/or
encapsulated within, surrounded by, and/or dispersed throughout an
aggregate of non-polymeric components.
[0227] Particles (e.g. nanoparticles, microparticles) may be
prepared using any method known in the art. For example,
particulate formulations can be formed by methods as
nanoprecipitation, flow focusing fluidic channels, spray drying,
single and double emulsion solvent evaporation, solvent extraction,
phase separation, milling, microemulsion procedures,
microfabrication, nanofabrication, sacrificial layers, simple and
complex coacervation, and other methods well known to those of
ordinary skill in the art. Alternatively or additionally, aqueous
and organic solvent syntheses for monodisperse semiconductor,
conductive, magnetic, organic, and other nanoparticles have been
described (Pellegrino et al., 2005, Small, 1:48; Murray et al.,
2000, Ann. Rev. Mat. Sci., 30:545; and Trindade et al., 2001, Chem.
Mat., 13:3843).
[0228] Methods for making microparticles for delivery of
encapsulated agents are described in the literature (see, e.g.,
Doubrow, Ed., "Microcapsules and Nanoparticles in Medicine and
Pharmacy," CRC Press, Boca Raton, 1992; Mathiowitz et al., 1987, J.
Control. Release, 5:13; Mathiowitz et al., 1987, Reactive Polymers,
.delta.: 275; and Mathiowitz et al., 1988, J. Appl. Polymer Sci.,
35:755).
[0229] In some embodiments, the targeting moiety comprises an
nucleic acid targeting moiety.
[0230] In general, a nucleic acid targeting moiety is any
polynucleotide that binds to a component associated with an organ,
tissue, cell, extracellular matrix component, and/or intracellular
compartment (the target).
[0231] In some embodiments, the nucleic acid targeting moieties are
aptamers.
[0232] An aptamer is typically a polynucleotide that binds to a
specific target structure that is associated with a particular
organ, tissue, cell, extracellular matrix component, and/or
intracellular compartment. In general, the targeting function of
the aptamer is based on the three-dimensional structure of the
aptamer. In some embodiments, binding of an aptamer to a target is
typically mediated by the interaction between the two- and/or
three-dimensional structures of both the aptamer and the target. In
some embodiments, binding of an aptamer to a target is not solely
based on the primary sequence of the aptamer, but depends on the
three-dimensional structure(s) of the aptamer and/or target. In
some embodiments, aptamers bind to their targets via complementary
Watson-Crick base pairing which is interrupted by structures (e.g.
hairpin loops) that disrupt base pairing.
[0233] In some embodiments, the nucleic acid targeting moieties are
spiegelmers (PCT Publications WO 98/08856, WO 02/100442, and WO
06/117217). In general, spiegelmers are synthetic, mirror-image
nucleic acids that can specifically bind to a target (i.e. mirror
image aptamers). Spiegelmers are characterized by structural
features which make them not susceptible to exo- and
endo-nucleases.
[0234] One of ordinary skill in the art will recognize that any
nucleic acid targeting moiety (e.g. aptamer or spiegelmer) that is
capable of specifically binding to a target can be used in
accordance with the present invention. In some embodiments, nucleic
acid targeting moieties to be used in accordance with the present
invention may target a marker associated with a disease, disorder,
and/or condition. In some embodiments, nucleic acid targeting
moieties to be used in accordance with the present invention may
target cancer-associated targets. In some embodiments, nucleic acid
targeting moieties to be used in accordance with the present
invention may target tumor markers. Any type of cancer and/or any
tumor marker may be targeted using nucleic acid targeting moieties
in accordance with the present invention. To give but a few
examples, nucleic acid targeting moieties may target markers
associated with prostate cancer, lung cancer, breast cancer,
colorectal cancer, bladder cancer, pancreatic cancer, endometrial
cancer, ovarian cancer, bone cancer, esophageal cancer, liver
cancer, stomach cancer, brain tumors, cutaneous melanoma, and/or
leukemia.
[0235] Nucleic acids of the present invention (including nucleic
acid nucleic acid targeting moieties and/or functional RNAs to be
delivered, e.g., RNAi-inducing entities, ribozymes, tRNAs, etc.,
described in further detail below) may be prepared according to any
available technique including, but not limited to chemical
synthesis, enzymatic synthesis, enzymatic or chemical cleavage of a
longer precursor, etc. Methods of synthesizing RNAs are known in
the art (see, e.g., Gait, M. J. (ed.) Oligonucleotide synthesis: a
practical approach. Oxford [Oxfordshire], Washington, D.C.: IRL
Press, 1984; and Herdewijn, P. (ed.) Oligonucleotide synthesis:
methods and applications, Methods in molecular biology, v. 288
(Clifton, N.J.) Totowa, N.J.: Humana Press, 2005).
[0236] The nucleic acid that forms the nucleic acid nucleic acid
targeting moiety may comprise naturally occurring nucleosides,
modified nucleosides, naturally occurring nucleosides with
hydrocarbon linkers (e.g., an alkylene) or a polyether linker
(e.g., a PEG linker) inserted between one or more nucleosides,
modified nucleosides with hydrocarbon or PEG linkers inserted
between one or more nucleosides, or a combination of thereof. In
some embodiments, nucleotides or modified nucleotides of the
nucleic acid nucleic acid targeting moiety can be replaced with a
hydrocarbon linker or a poly ether linker provided that the binding
affinity and selectivity of the nucleic acid nucleic acid targeting
moiety is not substantially reduced by the substitution (e.g., the
dissociation constant of the nucleic acid nucleic acid targeting
moiety for the target should not be greater than about
1.times.10.sup.-3 M).
[0237] It will be appreciated by those of ordinary skill in the art
that nucleic acids in accordance with the present invention may
comprise nucleotides entirely of the types found in naturally
occurring nucleic acids, or may instead include one or more
nucleotide analogs or have a structure that otherwise differs from
that of a naturally occurring nucleic acid. U.S. Pat. Nos.
6,403,779; 6,399,754; 6,225,460; 6,127,533; 6,031,086; 6,005,087;
5,977,089; and references therein disclose a wide variety of
specific nucleotide analogs and modifications that may be used. See
Crooke, S. (ed.) Antisense Drug Technology: Principles, Strategies,
and Applications (1st ed), Marcel Dekker; ISBN: 0824705661; 1st
edition (2001) and references therein. For example,
2'-modifications include halo, alkoxy and allyloxy groups. In some
embodiments, the 2'-OH group is replaced by a group selected from
H, OR, R, halo, SH, SR, NH2, NHR, NR2 or CN, wherein R is C1-C6
alkyl, alkenyl, or alkynyl, and halo is F, Cl, Br, or I. Examples
of modified linkages include phosphorothioate and
5'-N-phosphoramidite linkages.
[0238] Nucleic acids comprising a variety of different nucleotide
analogs, modified backbones, or non-naturally occurring
internucleoside linkages can be utilized in accordance with the
present invention.
[0239] Nucleic acids of the present invention may include natural
nucleosides (i.e., adenosine, thymidine, guanosine, cytidine,
uridine, deoxyadenosine, deoxythymidine, deoxyguanosine, and
deoxycytidine) or modified nucleosides. Examples of modified
nucleotides include base modified nucleoside (e.g., aracytidine,
inosine, isoguanosine, nebularine, pseudouridine,
2,6-diaminopurine, 2-aminopurine, 2-thiothymidine,
3-deaza-5-azacytidine, 2'-deoxyuridine, 3-nitorpyrrole,
4-methylindole, 4-thiouridine, 4-thiothymidine, 2-aminoadenosine,
2-thiothymidine, 2-thiouridine, 5-bromocytidine, 5-iodouridine,
inosine, 6-azauridine, 6-chloropurine, 7-deazaadenosine,
7-deazaguanosine, 8-azaadenosine, 8-azidoadenosine, benzimidazole,
M1-methyladenosine, pyrrolo-pyrimidino, 2-amino-6-chloropurine,
3-methyl adenosine, 5-propynylcytidine, 5-propynyluridine,
5-bromouridine, 5-fluorouridine, 5-methylcytidine,
7-deazaadenosine, 7-deazaguanosine, 8-oxoadenosine, 8-oxoguanosine,
O(6)-methylguanine, and 2-thiocytidine), chemically or biologically
modified bases (e.g., methylated bases), modified sugars (e.g.,
2'-fluororibose, 2'-aminoribose, 2'-azidoribose, 2'-O-methylribose,
L-enantiomeric nucleosides arabinose, and hexose), modified
phosphate groups (e.g., phosphorothioates and 5'-N-phosphoramidite
linkages), and combinations thereof. Natural and modified
nucleotide monomers for the chemical synthesis of nucleic acids are
readily available. In some cases, nucleic acids comprising such
modifications display improved properties relative to nucleic acids
consisting only of naturally occurring nucleotides. In some
embodiments, nucleic acid modifications described herein are
utilized to reduce and/or prevent digestion by nucleases (e.g.
exonucleases, endonucleases, etc.). For example, the structure of a
nucleic acid may be stabilized by including nucleotide analogs at
the 3' end of one or both strands order to reduce digestion.
[0240] Modified nucleic acids need not be uniformly modified along
the entire length of the molecule. Different nucleotide
modifications and/or backbone structures may exist at various
positions in the nucleic acid. One of ordinary skill in the art
will appreciate that the nucleotide analogs or other
modification(s) may be located at any position(s) of a nucleic acid
such that the function of the nucleic acid is not substantially
affected. To give but one example, modifications may be located at
any position of a nucleic acid targeting moiety such that the
ability of the nucleic acid targeting moiety to specifically bind
to the target is not substantially affected. The modified region
may be at the 5'-end and/or the 3'-end of one or both strands. For
example, modified nucleic acid targeting moieties in which
approximately 1-5 residues at the 5' and/or 3' end of either of
both strands are nucleotide analogs and/or have a backbone
modification have been employed. The modification may be a 5' or 3'
terminal modification. One or both nucleic acid strands may
comprise at least 50% unmodified nucleotides, at least 80%
unmodified nucleotides, at least 90% unmodified nucleotides, or
100% unmodified nucleotides.
[0241] Nucleic acids in accordance with the present invention may,
for example, comprise a modification to a sugar, nucleoside, or
internucleoside linkage such as those described in U.S. Patent
Application Publications 2003/0175950, 2004/0192626, 2004/0092470,
2005/0020525, and 2005/0032733. The present invention encompasses
the use of any nucleic acid having any one or more of the
modification described therein. For example, a number of terminal
conjugates, e.g., lipids such as cholesterol, lithocholic acid,
aluric acid, or long alkyl branched chains have been reported to
improve cellular uptake. Analogs and modifications may be tested
using, e.g., using any appropriate assay known in the art, for
example, to select diose that result in improved delivery of a
therapeutic or diagnostic agent, improved specific binding of an
nucleic acid targeting moiety to a target, etc. In some
embodiments, nucleic acids in accordance with the present invention
may comprise one or more non-natural nucleoside linkages. In some
embodiments, one or more internal nucleotides at the 3'-end,
5'-end, or both 3'- and 5'-ends of the nucleic acid targeting
moiety are inverted to yield a linkage such as a 3'-3' linkage or a
5'-5' linkage.
[0242] In some embodiments, nucleic acids in accordance with the
present invention are not synthetic, but are naturally-occurring
entities that have been isolated from their natural
environments.
[0243] Any method can be used to design novel nucleic acid
targeting moieties (see, e.g., U.S. Pat. Nos. 6,716,583; 6,465,189;
6,482,594; 6,458,543; 6,458,539; 6,376,190; 6,344,318; 6,242,246;
6,184,364; 6,001,577; 5,958,691; 5,874,218; 5,853,984; 5,843,732;
5,843,653; 5,817,785; 5,789,163; 5,763,177; 5,696,249; 5,660,985;
5,595,877; 5,567,588; and 5,270,163; and U.S. Patent Application
Publications 2005/0069910, 2004/0072234, 2004/0043923,
2003/0087301, 2003/0054360, and 2002/0064780). The present
invention provides methods for designing novel nucleic acid
targeting moieties. The present invention further provides methods
for isolating or identifying novel nucleic acid targeting moieties
from a mixture of candidate nucleic acid targeting moieties.
[0244] Nucleic acid targeting moieties that bind to a protein, a
carbohydrate, a lipid, and/or a nucleic acid can be designed and/or
identified. In some embodiments, nucleic acid targeting moieties
can be designed and/or identified for use in the complexes of the
invention that bind to proteins and/or characteristic portions
thereof, such as tumor-markers, integrins, cell surface receptors,
transmembrane proteins, intercellular proteins, ion channels,
membrane transporter proteins, enzymes, antibodies, chimeric
proteins etc. In some embodiments, nucleic acid targeting moieties
can be designed and/or identified for use in the complexes of the
invention that bind to carbohydrates and/or characteristic portions
thereof, such as glycoproteins, sugars (e.g., monosaccharides,
disaccharides and polysaccharides), glycocalyx (i.e., the
carbohydrate-rich peripheral zone on the outside surface of most
eukaryotic cells) etc. In some embodiments, nucleic acid targeting
moieties can be designed and/or identified for use in the complexes
of the invention that bind to lipids and/or characteristic portions
thereof, such as oils, saturated fatty acids, unsaturated fatty
acids, glycerides, hormones, steroids (e.g., cholesterol, bile
acids), vitamins (e.g. vitamin E), phospholipids, sphingolipids,
lipoproteins etc. In some embodiments, nucleic acid targeting
moieties can be designed and/or identified for use in the complexes
of the invention that bind to nucleic acids and/or characteristic
portions thereof, such as DNA nucleic acids; RNA nucleic acids;
modified DNA nucleic acids; modified RNA nucleic acids; and nucleic
acids that include any combination of DNA, RNA, modified DNA, and
modified RNA; etc.
[0245] Nucleic acid targeting moieties (e.g. aptamers or
spiegelmers) may be designed and/or identified using any available
method. In some embodiments, nucleic acid targeting moieties are
designed and/or identified by identifying nucleic acid targeting
moieties from a candidate mixture of nucleic acids. Systemic
Evolution of Ligands by Exponential Enrichment (SELEX), or a
variation thereof, is a commonly used method of identifying nucleic
acid targeting moieties that bind to a target from a candidate
mixture of nucleic acids.
[0246] Nucleic acid targeting moieties that bind selectively to any
target can be isolated by the SELEX process, or a variation
thereof, provided that the target can be used as a target in the
SELEX process.
B. Immunotherapeutics
[0247] In general, the combination or composition of the present
invention comprises an immunotherapeutic.
[0248] By "immunotherapeutics" herein is meant a compound, a
molecule, or an agent that is capable of stimulating or enhancing
the body's immune system or tumor cells. Immunotherapeutics are
used for the treatment of disease by inducing, enhancing, or
suppressing an immune response. Immunotherapeutics of the present
invention generally are designed to elicit or amplify an immune
response, rather than suppress an immune response.
[0249] In general, the immunotherapeutics of the present invention
act, directly or indirectly, on toll like receptors,
nucleotide-oligomerization domain-like receptors, RIG-I-Like
receptors, c-type lectin receptors, or cytosolic DNA Sensors, or a
combination thereof. Particually, the immunotherapeutics of the
present invention are capable of activating a human plasmacytoid
dendritic cell, myeloid dendritic cell, NK cell, or tumor cell, or
a combination thereof.
[0250] In some embodiments, the immunotherapeutics of the present
invention activate human immune cells, including but not limited to
dendritic cells, macrophages, monocytes, myeloid-derived suppressor
cells, NK cells, B cells, T cells, or tumor cells, or a combination
thereof.
[0251] Dendritic cells are the most powerful antigen-presenting
cells. Dendritic cells play an essential role for the initiation of
both innate and adaptive immune responses. Dendritic cells also
play a key role in the induction and maintenance of immune
tolerance.
[0252] By "dendritic cells" (DC) herein is meant a heterogeneous
cell population including two main subtypes: namely, myeloid DC
(mDC) and plasmacytoid DC (pDC) (Steinman et al., 1979, J. Exp.
Med., 149, 1-16). These two blood DC subsets were originally
differentiated by their expression of CD11c (integrin complement
receptor) and CD123 (IL-3R.alpha.). Each of the pDC and mDC
populations constitutes between about 0.2 to about 0.6% of the PBMC
population in humans.
[0253] By "pDC" herein is meant plasmacytoid dendritic cells and
they represent a subtype of dendritic cells found in the blood and
peripheral lymphoid organs. These cells express the surface markers
CD123, BDCA-2(CD303) and BDCA-4(CD304) and HLA-DR, but do not
express CD11c, CD14, CD3, CD20 or CD56, which distinguishes them
from conventional dendritic cells, monocytes, T-cells, B cells and
NK cells. As components of the innate immune system, these cells
express intracellular Toll-like receptors 7 and 9, which enable the
detection of viral and bacterial nucleic acids, such as ssRNA or
CpG DNA motifs. Upon stimulation and subsequent activation, these
cells produce large amounts of Type I interferon (mainly
IFN-.alpha. and IFN-.beta.) and Type III interferon (e.g.,
IFN-.gamma.), which are critical pleiotropic anti-viral compounds
mediating a wide range of effects. By generating a large number of
type I interferon, cytokines and chemokines, plasmacytoid dendritic
cells are widely involved in the body's innate and adaptive immune
responses. They can regulate NK cells, T cells, B cells and other
cells involved in immune response intensity, duration, and response
mode, thus play a very important function in tumor, infection and
autoimmune disease. (Liu Y J. IPC: professional type 1
interferon-producing cells and plasmacytoid dendritic cell
precursors. Annu Rev Immunol. 2005; 23:275-306. Gilliet M, Cao W,
Liu Y J. Plasmacytoid dendritic cells: sensing nucleic acids in
viral infection and autoimmune diseases. Nat Rev Immunol. 2008
August; 8 (8):0.594-606).
[0254] By "mDC" herein is meant myeloid dendritic cells and they
represent a subtype of circulating dendritic cells found in blood
and peripheral lymphoid organs. These cells express the surface
markers CD11c, CD1a, HLA-DR and either BDCA-1 (CD1c) or BDCA-3
(CD141). They do not express BDCA-2 or CD123, which distinguishes
them from pDC. mDC also do not express CD3, CD20 or CD56. As
components of the innate immune system, mDC express Toll-like
receptors (TLR), including TLR2, 3, 4, 5, 6 and 8, which enable the
detection of bacterial and viral components. Upon stimulation and
subsequent activation, these cells are the most potent antigen
presenting cells to activate antigen-specific CD4 as well as CD8 T
cells. In addition, mDCs has the ability to produce large amounts
of IL-12 and IL23, which is critical for the induction of
Th1-mediated or Th17 cell-mediated immunity.
[0255] Study found that many solid tumors such as breast cancer and
head and neck cancer, ovarian cancer has pDC's invasion (Treilleux
I, Blay J Y, Bendriss-Vermare N et al. Dendritic cell infiltration
and prognosis of early stage breast cancer. Clin Cancer Res 2004;
10:7466-7474. Hartmann E, Wollenberg B, Rothenfusser S et al.
Identification and functional analysis of tumor-infiltrating
plasmacytoid dendritic cells in head and neck cancer. Cancer Res
2003; 63:6478-6487. Zou W P, Machelon V, Coulomb-L'Hermin A, et al.
Stromal-derived factor-1 in human tumors recruits and alters the
function of plasmacytoid precursor dendritic cells. Nat Med 2001;
7:1339-1346) and factors secreted by tumor cells inhibit DC
maturation. (Gabrilovich D I, Corak J, Ciemik I F et al. Decreased
antigen presentation by dendritic cells in patients with breast
cancer. Clin Cancer Res 1997; 3:483-490. Bell D, Chomarat P,
Broyles D et al. In breast carcinoma tissue, immature dendritic
cells reside within the tumor, whereas mature dendritic cells are
located in peritumoral areas. J Exp Med 1999; 190:1417-1425.
Menetrier-Caux C, Montmain G, Dieu M C et al. Inhibition of the
differentiation of dendritic cells from CD34 (+) progenitors by
tumor cells: role of interleukin-6 and macrophage
colony-stimulating factor. Blood 1998; 92:4778-4791). These
immature DC cells did not play a role in promoting anti-tumor
immunity. By contrast, DCs within the tumor microenvironment
promote tumor growth by inhibiting antitumor immunity and by
promoting angiogenesis. There is evidence that Toll-like receptor 7
agonist Imiquimod, and Toll-like receptor 9 agonist CpG drugs can
stimulate pDC within the tumor microenvironment to inhibit tumor
development. (Dummer R, Urosevic M, Kempf W et al. Imiquimod in
basal cell carcinoma: how does it work? Br J Dermatol 2003;
149:57-58. Miller R L, Gerster J F, Owens M L et al Imiquimod
applied topically: a novel immune response modifier and new class
of drag. Int J Immunopharmacol 1999; 21:1-14. Hofmann M A, Kors C,
Audring H et al Phase 1 evaluation of intralesionally injected
TLR9-agonist PF-3512676 in patients with basal cell carcinoma or
metastatic melanoma. J Immunother 2008; 31:520-527).
[0256] Natural killer (NK) cells are a type of cytotoxic lymphocyte
that constitutes a major component of the immune system. NK cells
are a subset of peripheral blood lymphocytes defined by the
expression of CD56 or CD 16 and the absence of the T cell receptor
(CD3). They recognize and kill transformed cell lines without
priming in an MHC-unrestricted fashion. NK cells play a major role
in the rejection of tumors and cells infected by viruses. The
process by which an NK cell recognizes a target cell and delivers a
sufficient signal to trigger target lysis is determined by an array
of inhibitory and activating receptors on the cell surface. NK
discrimination of self from altered self involves inhibitory
receptor recognition of MHC-I molecules and non-MHC ligands like
CD48 and Clr-1b. NK recognition of infected or damaged cells
(altered self) is coordinated through stress induced ligands (e.g.,
MICA, MICE, Rae1, H60, Mult1) or vitally encoded ligands (e.g.,
m157, hemaglutinin) recognized by various activating receptors,
including NKG2D, Ly49H and NKp46/Ncr1.
[0257] NK cells represent the predominant lymphoid cell in the
peripheral blood for many months after allogeneic or autologous
stem cell transplant and they have a primary role in immunity to
pathogens during this period (Reittie et al (1989) Blood 73:
1351-1358; Lowdell et al (1998) Bone Marrow Transplant 21:
679-686). The role of NK cells in engraftment, graft-versus-host
disease, anti-leukemia activity and post-transplant infection is
reviewed in Lowdell (2003) Transfusion Medicine 13:399-404.
[0258] Human NK cells mediate the lysis of tumor cells and
virus-infected cells via natural cytotoxicity and
antibody-dependent cellular cytotoxicity (ADCC).
[0259] Human NK cells are controlled by positive and negative
cytolytic signals. Negative (inhibitory) signals are transduced by
C-lectin domain containing receptors CD94/NKG2A and by some Killer
Immunoglobulin-like Receptors (KIRs). The regulation of NK lysis by
inhibitory signals is known as the "missing self" hypothesis in
which specific HLA-class I alleles expressed on the target cell
surface ligate inhibitory receptors on NK cells. The
down-regulation of HLA molecules on tumor cells and some vitally
infected cells (e.g. CMV) lowers this inhibition below a target
threshold and the target cells may become susceptible to NK
cell-mediated lysis if the target cells also carry NK-priming and
activating molecules. TLR7, TLR8 or TLR9 agonists can activate both
mDC and pDCs to produce type I IFNs and express costimulatory
molecules such as GITR-ligand, which subsequently activate NK cells
to produce IFN-g and potently promote NK cell killing function.
[0260] Inhibitory receptors fall into two groups, those of the
Ig-superfamily called Killer Immunoglobulin-like Receptors (KIRs)
and those of the lectin family, the NKG2, which form dimers with
CD94 at the cell surface. KIRs have a 2- or 3-domain extracellular
structure and bind to HLA-A, -B or -C. The NKG2/CD94 complexes
ligate HLA-E.
[0261] Inhibitory KIRs have up to 4 intracellular domains which
contain ITIMs and the best characterized are KIR2DL1, KIR2DL2 and
KIR2DL3 which are known to bind HLA-C molecules. KIR2DL2 and
KIR2DL3 bind the group 1 HLA-C alleles while KIR2DL1 binds to group
2 alleles. Certain leukemia/lymphoma cells express both group 1 and
2 HLA-C alleles and are known to be resistant to NK-mediated cell
lysis.
[0262] With regards to positive activating signals, ADCC is thought
to be mediated via CD 16, and a number of triggering receptors
responsible for natural cytotoxicity have been identified,
including CD2, CD38, CD69, NKRP-I, CD40, B7-2, NK-TR, NKp46, NKp30
and NKp44. In addition, several KIR molecules with short
intracytoplasmic tails are also stimulatory. These KIRs (KIR2DS1,
KIR2DS2 and KIR2DS4) are known to bind to HLA-C; their
extracellular domains being identical to their related inhibitory
KIRs. The activatory KIRs lack the ITIMs and instead associate with
DAP 12 leading to NK cell activation. The mechanism of control of
expression of inhibitory versus activatory KIRs remains
unknown.
[0263] Several reports have described the expression of TLRs in
mouse or human cancer or cancer cell lines. For example, TLR1 to
TLR6 are expressed by colon, lung, prostate, and melanoma mouse
tumor cell lines (Huang B, et al. Toll-like receptors on tumor
cells facilitate evasion of immune surveillance. Cancer Res. 2005;
65(12):5009-5014), TLR3 is expressed in human breast cancer cells
(Salaun B, Coste I, Rissoan M C, Lebecque S J, Renno T. TLR3 can
directly trigger apoptosis in human cancer cells. J Immunol. 2006;
176(8):4894-4901), hepatocarcinoma and gastric carcinoma cells
express TLR2 and TLR4 (Huang B, et al. Listeria monocytogenes
promotes tumor growth via tumor cell toll-like receptor 2
signaling. Cancer Res. 2007; 67(9):4346-4352), and TLR9 (Droemann
D, et al. Human lung cancer cells express functionally active
Toll-like receptor 9. Respir Res. 2005; 6:1.) and TLR4 (He W, Liu
Q, Wang L, Chen W, Li N, Cao X. TLR4 signaling promotes immune
escape of human lung cancer cells by inducing immunosuppressive
cytokines and apoptosis resistance. Mol Immunol. 2007;
44(11):2850-2859.) are expressed by human lung cancer cells. TLR7
and TLR8 are found in tumor cells of human lung cancer
(Cherfils-Vicini J, Platonova S, Gillard M, Laurans L, Validire P,
Caliandro R, Magdeleinat P, Mami-Chouaib F, Dieu-Nosjean M C,
Fridman W H, Damotte D, Sautes-Fridman C, Cremer I. J. Clin Invest.
2010; 120(4): 1285-1297).
[0264] TLR are a family of proteins that sense a microbial product
and/or initiates an adaptive immune response. TLRs activate a
dendritic cell (DC). TLRs are conserved membrane spanning molecules
containing an ectodomain of leucine-rich repeats, a transmembrane
domain and an intracellular TIR (Toll/interleukin receptor) domain.
TLRs recognize distinct structures in microbes, often referred to
as "PAMPs" (pathogen associated molecular patterns). Ligand binding
to TLRs invokes a cascade of intracellular signaling pathways that
induce the production of factors involved in inflammation and
immunity.
[0265] In some embodiments, the immunotherapeutic is a TLR7 and/or
TLR8 agonist. TLR7 and TLR8 are phylogenetically and structurally
related. TLR7 is selectively expressed by human pDCs and B cells.
TLR8 is predominantly expressed mDCs, monocytes, macrophages and
myeloid suppressor cells. TLR7-specific agonists activate
plasmacytoid DCs (pDCs) to produce large amounts of type 1 IFNs and
expressing high levels of costimulatory molecules that promote
activation of T cells, NK cells, B cells and mDCs. TLR8-specific
agonists activate myeloid DCs, monocytes, macrophages or
myeloid-derived suppressor cells to produce large amounts of type 1
IFN, IL-12 and IL-23, and express high levels of MHC class I, MHC
class II and costimulatory molecules that promote the activation of
antigen specific CD4 and CD8+ T cells.
[0266] In some embodiments, the immunotherapeutic is a TLR7 and/or
TLR8 agonist that is represented by the structure of Formula
(I):
##STR00003##
wherein dashed line represents bond or absence of bond; X is S or
--NR.sub.1, R.sub.1 is -W.sub.0-W.sub.1-W.sub.2-W.sub.3-W.sub.4,
W.sub.0 is a bond, alkyl alkenyl, alkynyl, alkoxy, or
-alkyl-S-alkyl-, W.sub.1 is a bond, --O--, or --NR.sub.2--, wherein
R.sub.2 is hydrogen, alkyl or alkenyl, W.sub.2 is a bond, --O--,
--C(O)--, --C(S)--, or --S(O).sub.2--, W.sub.3 is a bond,
--NR.sub.3--, wherein R.sub.3 is hydrogen, alkyl or alkenyl,
W.sub.4 is hydrogen, alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl,
aryl, aryloxy, heteroaryl, or heterocyclyl, each of which is
optionally substituted by one or more substituents selected from
the group consisting of hydroxyl, alkoxy, alkyl, alkenyl, alkynyl,
cycloalkyl, aryl, heteroaryl, heterocyclyl. --NH.sub.2, nitro,
-alkyl-hydroxyl, -alkyl-aryl, -alkyl-heteroaryl,
-alkyl-heterocyclyl, --O--R.sub.4, --O-alkyl-R.sub.4,
-alkyl-O--R.sub.4, --C(O)--R.sub.4, -alkyl-C(O)--R.sub.4,
-alkyl-C(O)--O--R.sub.4, --C(O)--O--R.sub.4, --S--R.sub.4,
--S(O).sub.2--R.sub.4, --NH--S(O).sub.2--R.sub.4, -alkyl-S--R4,
-alkyl-S(O).sub.2--R.sub.4, --NHR.sub.4, --NR.sub.4R.sub.4,
--NH-alkyl-R.sub.4, halogen, --CN, --NO.sub.2, and --SH, wherein
R.sub.4 is independently hydrogen, alkyl, alkenyl, -alkyl-hydroxyl,
aryl, heteroaryl, heterocyclyl, or haloalkyl; Z is hydrogen, alkyl,
alkenyl, alkynyl, alkoxy, aryl, haloalkyl, heteroaryl,
heterocyclyl, each of which can be optionally substituted by one or
more substituents selected from the group consisting of hydroxyl,
alkoxy, alkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclyl,
halogen, cyano, nitro, --N(R.sub.5).sub.2, -alkoxy-alkyl,
-alkoxy-alkenyl, --C(O)-alkyl, --C(O)--O-alkyl, --O--C(O)-alkyl,
--C(O)--N(R.sub.5).sub.2, aryl, heteroaryl, --CO-aryl, and
--CO-heteroaryl, wherein each R.sub.5 is independently hydrogen,
alkyl, haloalkyl, -alkyl-aryl, or -alkyl-heteroaryl; R is hydrogen,
alkyl, alkoxy, haloalkyl, halogen, aryl, heteroaryl, heterocyclyl,
each of which is optionally substituted by one or more substituents
selected from the group consisting of hydroxyl, alkoxy, alkyl,
alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, heterocyclyl,
--NH.sub.2, nitro, -alkyl-hydroxyl, -alkyl-aryl, -alkyl-heteroaryl,
-alkyl-heterocyclyl. --O--R.sub.4, --O-alkyl-R.sub.4,
-alkyl-O--R.sub.4, --C(O)--R.sub.4, --C(O)--NH--R.sub.4,
--C(O)--NR.sub.4R.sub.4, -alkyl-C(O)--R.sub.4,
-alkyl-C(O)--O--R.sub.4, --C(O)--O--R.sub.4, --O--C(O)--R.sub.4,
--S--R.sub.4, --C(O)--S--R.sub.4, --S--C(O)--R.sub.4,
--S(O).sub.2--R.sub.4, --NH--S(O).sub.2--R.sub.4,
-alkyl-S--R.sub.4, -alkyl-S(O).sub.2--R.sub.4, --NHR.sub.4,
--NR.sub.4R.sub.4, --NH-alkyl-R.sub.4, halogen, --CN, and --SH,
wherein R.sub.4 is independently hydrogen, alkyl, alkenyl, alkoxy,
-alkyl-hydroxyl, aryl, heteroaryl, heterocyclyl, or haloalkyl; n is
0, 1, 2, 3, or 4; Y is --NR.sub.6R.sub.7, --CR.sub.6R.sub.7R.sub.8,
or -alkyl-NH.sub.2, each of which can be optionally substituted by
one or more substituents selected from the group consisting of
hydroxyl, alkoxy, alkyl, alkenyl, alkynyl, --NH.sub.2, halogen,
--N(R.sub.5).sub.2, -alkoxyalkyl, -alkoxy-alkenyl, --C(O)-alkyl,
--C(O)--O-alkyl, --C(O)--N(R.sub.5).sub.2, aryl, heteroaryl,
--CO-aryl, and --CO-heteroaryl. wherein R.sub.6, R.sub.7 and
R.sub.8 are independently hydrogen, alkyl, alkenyl, alkoxy,
alkylamino, dialkylamino, alkylthio, arylthio, -alkyl-hydroxyl,
-alkyl-C(O)--O--R.sub.9, -alkyl-C(O)--R.sub.9, or
-alkyl-O--C(O)--R.sub.9, wherein each R.sub.5 is independently
hydrogen, alkyl, haloalkyl, -alkyl-aryl, or -alkyl-heteroaryl,
wherein R.sub.9 is hydrogen, alkyl, alkenyl, halogen, or haloalkyl;
X and Z taken together may optionally form a (5-9)-membered ring;
or a pharmaceutically acceptable salt or solvate thereof.
[0267] In some embodiments, X of Formula (I) is S.
[0268] In some embodiments, X of Formula (I) is --NR.sub.1, R.sub.1
is alkyl, -alkyl-W.sub.4, -alkyl-O--W.sub.4,
-alkyl-NH--C(O)--W.sub.4, -alkoxy-NH--C(O)--W.sub.4,
-alkyl-NH--C(O)--NH--W.sub.4, -alkoxy-NH--C(O)--NH--W.sub.4,
-alkyl-S(O).sub.2--W.sub.4, or -alkyl-NH--C(S)--W.sub.4, wherein
W.sub.4 is defined above.
[0269] In some embodiments, Z of Formula (I) is hydrogen, alkyl,
alkoxy, aryl, heteroaryl, haloalkyl, each of which is optionally
substituted by one to three substituents selected from the group
consisting of hydroxyl, alkyl, aryl, heteroaryl, heterocyclyl,
cyano, -alkoxy-alkyl, nitro, and --N(R.sub.5).sub.2, wherein each
R.sub.5 is independently hydrogen, alkyl, haloalkyl, -alkyl-aryl,
or -alkyl-heteroaryl.
[0270] In some embodiments, Y of Formula (I) is --NH.sub.2,
-alkyl-NH.sub.2, each of which is optionally substituted by one to
three substituents selected from the group consisting of alkyl,
alkoxy, alkenyl, and alkynyl.
[0271] In some embodiments, n of Formula (I) is 1 or 2.
[0272] In some embodiments, R of Formula (I) is aryl or heteroaryl
each of which is optionally substituted by one to three
substituents selected from the group consisting of hydroxyl,
alkoxy, -alkyl-hydroxyl, --O--R.sub.4, --O-alkyl-R.sub.4,
-alkyl-O--R.sub.4, --C(O)--R.sub.4, --C(O)--NH--R.sub.4,
--C(O)--NR.sub.4R.sub.4, -alkyl-C(O)--R.sub.4,
-alkyl-C(O)--O--R.sub.4, --C(O)--O--R.sub.4, --O--C(O)--R.sub.4,
--S--R.sub.4, --C(O)--S--R.sub.4, --S--C(O)--R.sub.4,
--S(O).sub.2--R.sub.4, --NH--S(O).sub.2--R.sub.4,
-alkyl-S--R.sub.4, -alkyl-S(O).sub.2--R.sub.4, --NHR.sub.4,
--NR.sub.4R.sub.4, --NH-alkyl-R.sub.4, halogen, --CN, and --SH,
wherein R is independently hydrogen, alkyl, alkenyl, alkoxy,
-alkyl-hydroxyl, aryl, heteroaryl, heterocyclyl, or haloalkyl.
[0273] In some embodiments, the immunotherapeutic is a TLR7 and/or
TLR8 agonist that is selected from Table 2. The compounds in Table
2 are described and characterized in more details in U.S. Pat. No.
4,689,338, U.S. Pat. No. 5,389,640, U.S. Pat. No. 5,226,575, U.S.
Pat. No. 6,110,929, U.S. Pat. No. 6,194,425, U.S. Pat. No.
5,352,784. U.S. Pat. No. 6,331,539, U.S. Pat. No. 5,482,936, U.S.
Pat. No. 6,451,810, WO2002/46192, WO2002/46193, WO2002/46194,
US2004/0014779 and US2004/0162309.
TABLE-US-00001 TABLE 2 Representative TLR7 and/or TLR8 Agonists
Name Structure 2-propylthiazolo[4,5- c]quinolin-4-amine (CL075)
##STR00004## 1-(2-methylpropyl)-1H- imidazo[4,5-c]quinolin-4- amine
(Imiquimod) ##STR00005## 4-amino-2- (ethoxymethyl)-a,a-di-
methyl-1H-imidazo[4,5- c]quinoline-1-ethanol (Resiquimod)
##STR00006## 1-(4-amino-2- ethylaminomethylimidazo-
[4,5-c]quinolin-1-yl)-2- methylpropan-2-ol (Gardiquimod)
##STR00007## N-[4-(4-amino-2-ethyl- 1H-imidazo[4,5-
c]quinolin-1-yl)butyl-] methanesulfonamide (CM001) ##STR00008##
7-allyl-7,8-dihydro-8- oxo-guanosine (Loxoribine) ##STR00009##
4-amino-2-ethoxymethyl- aa-dimethyl-6,7,8,9- tetrahydro-1h-
imidazo[4,5-c]quinoline- 1-ethanol ol ##STR00010##
4-amino-aa-dimethyl-2- methoxyethyl-1h- imidazo[4,5-
c]quinoline-1-ethanol ##STR00011## 1-(2-(3-
(benzyloxy)propoxy)ethyl)- 2-(ethoxymethyl)-1H-
imidazo[4,5-c]quinolin-4- amine ##STR00012## N-[4-(4-amino-2-butyl-
1H-imidazo[4,5- c][1,5]naphthyridin-1- yl)butyl]-n'-butylurea
##STR00013## N1-[2-(4-amino-2-butyl- 1H-imidazo[4,5-c][1,5]
naphthyridin-1- yl)ethyl]-2-amino-4- methylpentanamide ##STR00014##
N-(2-{2-[4-amino-2-(2- methoxyethyl)-1H- imidazo[4,5-c]quinolin-
1-yl]ethoxy}ethyl)-n'- phenylurea ##STR00015## 1-(2-amino-2-
methylpropyl)-2- (ethoxymethyl)-1H- imidazo[4,5-c]quinolin- 4-amine
##STR00016## l-{4-[(3,5- dichlorophenyl)sulfonyl] butyl}-2-ethyl-
1H-imidazo[4,5- c]quinolin-4-amine ##STR00017## N-(2-{2-[4-amino-2-
(ethoxymethyl)-1H- imidazo[4,5- c]quinolin-1- yl]ethoxy}ethyl)-n'-
cyclohexylurea ##STR00018## N-{3-[4-amino-2- (ethoxymethyl)-1H-
imidazo[4,5- c]quinolin-1-yl]propyl}- n'-(3- cyanophenyl)thiourea
##STR00019## N-[3-(4-amino-2-butyl- 1H-imidazo[4,5- c]quinolin-1-
yl)-2,2-dimethylpropyl] benzamide ##STR00020## 2-butyl-1-[3-
(methylsulfonyl)propyl]- 1H-imidazo[4,5-c] quinolin-4-amine
##STR00021## N-{2-[4-amino-2- (ethoxymethyl)-1H- imidazo[4,5-
c]quinolin-1-yl]-1,1- dimethylethyl}-2- ethoxyacetamide
##STR00022## 1-[4-amino-2- ethoxymethyl-7-(pyridin- 4-yl)-1H-
imidazo[4,5-c]quinolin- 1-yl]-2-methylpropan-2- ol ##STR00023##
1-[4-amino-2- (ethoxymethyl)-7- (pyridin-3-yl)-1H-
imidazo[4,5-c]quinolin- 1-yl]-2-methylpropan-2- ol ##STR00024##
N-{3-[4-amino-1-(2- hydroxy-2- methylpropyl)-2- (methoxyethyl)-1H-
imidazo[4,5-c]quinolin-7- yl]phenyl}methane- sulfonamide
##STR00025## 1-[4-amino-7-(5- hydroxymethylpyridin-3- yl)-2-(2-
methoxyethyl)-1H- imidazo[4,5-c]quinolin-1- yl]-2-methylpropan-2-ol
##STR00026## 3-[4-amino-2- (ethoxymethyl)-7- (pyridin-3-yl)-1H-
imidazo[4,5-c]quinolin- 1-yl]propane-1,2-diol ##STR00027##
1-[2-(4-amino-2- ethoxymethyl-1H- imidazo[4,5- c]quinolin-1-yl)-1,1
- dimethylethyl]-3- propylurea ##STR00028## 1-[2-(4-amino-2-
ethoxymethyl-1H- imidazo[4,5- c]quinolin-1-yl)-1,1-
dimethylethyl]-3- cyclopentylurea ##STR00029##
1-[(2,2-dimethyl-1,3- dioxolan-4-yl)methyl]-2- (ethoxymethyl)-7-(4-
hydroxymethylphenyl)- 1H-imidazo[4,5-c] quinolin-4-amine
##STR00030## 4-[4-amino-2- ethoxymethyl-1-(2- hydroxy-2-
methylpropyl)-1H- imidazo[4,5-c]quinolin- 7-yl]-N- methoxy-N-
methylbenzamide ##STR00031## 2-ethoxymethyl-N1- isopropyl-6,7,8,9-
tetrahydro-1H- imidazo[4,5- c]quinoline-1,4-diamine ##STR00032##
1-[4-amino-2-ethyl-7- (pyridin-4-yl)-1H- imidazo[4,5-
c]quinolin-l-yl]-2- methylpropan-2-ol ##STR00033##
N-[4-(4-amino-2-ethyl- 1H-imidazo[4,5- c]quinolin-1-
yl)butyl]methane- sulfonamide ##STR00034## N-[4-(4-amino-2-butyl-
1H-imidazo[4,5- c][1,5]naphthyridin-1- yl)butyl]-n'- cyclohexylurea
##STR00035## 3M-34240 ##STR00036## 3M-052 ##STR00037## 3M-854A
##STR00038## ##STR00039## ##STR00040##
[0274] Preferably in some embodiments, the immunotherapeutic is
Resiquimod or Imiquimod.
[0275] In some embodiments, the immunotherapeutic is a TLR
modulator (e.g., TLR7 and/or TLR8 agonist) that is represented by
structure of Formula (II):
##STR00041##
wherein V is --NR.sub.6R.sub.7, wherein each of R.sub.6 and R.sub.7
is independently hydrogen, alkyl, alkenyl, alkoxy, alkylamino,
dialkylamino, alkylthio, arylthio, -alkyl-hydroxyl,
-alkyl-C(O)--O--R.sub.9, -alkyl-C(O)--R.sub.9, or
-alkyl-O--C(O)--R.sub.9, wherein R.sub.9 is hydrogen, alkyl,
alkenyl, hydrogen, or haloalkyl; R.sub.10 and R.sub.11 are
independently hydrogen, alkyl, alkenyl, aryl, haloalkyl,
heteroaryl, heterocyclyl, or cycloalkyl, each of which is
optionally substituted by one or more substituents selected from
the group consisting of hydroxyl, alkoxy, alkyl, alkenyl, alkynyl,
halogen. --N(R.sub.5).sub.2, -alkoxyalkyl, -alkoxy-alkenyl,
--C(O)-alkyl, --C(O)--O-alkyl, --C(O)--N(R.sub.5).sub.2, aryl,
heteroaryl, --CO-aryl, and --CO-heteroaryl, wherein each R.sub.5 is
independently hydrogen, alkyl, haloalkyl, -alkyl-aryl, or
-alkyl-heteroaryl, or a pharmaceutically acceptable salt or solvate
thereof.
[0276] In some embodiments, the immunotherapeutic is a TLR
modulator (e.g., TLR7 and/or TLR8 agonist) that is represented by
structure of Formula (III):
##STR00042##
wherein is a double bond or a single bond; R.sub.2 and R.sub.3 are
independently selected from H and lower alkyl, or R.sub.2 and
R.sub.3 are connected to form a saturated carbocycle having from 3
to 7 ring members; one of R.sub.7 and R.sub.8 is
##STR00043##
and the other is hydrogen; R.sub.4 is --NR.sub.cR.sub.d or
--OR.sub.10; R.sub.c and R.sub.d are lower alkyl, where the alkyl
is optionally substituted with one or more --OH; R.sub.10, is
alkyl, where the alkyl is optionally substituted with one or more
--OH; Z is C and is a double bond, or Z is N and is a single bond;
R.sub.a and R.sub.b are independently selected from H, alkyl,
alkenyl, alkynyl, and R.sub.e, wherein the alkyl is optionally
substituted with one or more --OR.sub.10, or R.sub.e, R.sub.e is
selected from --NH.sub.2, --NH(alkyl), and --N(alkyl).sub.2,
R.sub.1 is absent when is a double bond, or when is a single bond.
N.sub.1--R.sub.1 and one of R.sub.a or R.sub.b are connected to
form a saturated, partially unsaturated, or unsaturated heterocycle
having 5-7 ring members and the other of R.sub.a or R.sub.b may be
hydrogen or absent as necessary to accommodate ring unsaturation;
and at least one of the following A-D applies: A) R.sub.7 is not
hydrogen B) R.sub.8 is not hydrogen and at least one of R.sub.a and
R.sub.b is not hydrogen; C) Z is N; or D) N.sub.1--R.sub.1 and one
of R.sub.a or R.sub.b are connected to form a saturated, partially
unsaturated, or unsaturated heterocycle having 5-7 ring members. US
20140088085A1, the disclosure of which is incorporated by
references in its entirety.
[0277] In some embodiments, R.sub.7 of the compound of Formula
(III) is
##STR00044##
Additionally, at least one of R.sub.a and R.sub.b is not hydrogen
in the compound of Formula (III), or, for example, one of R.sub.a
and R.sub.b is alkyl and the other of R.sub.a and R.sub.b is
hydrogen. Further, the alkyl of Formula (III) is substituted with
R.sub.e. In a different embodiment, both R.sub.a and R.sub.b are
alkyl or, one of R.sub.a and R.sub.b is R.sub.e and the other
R.sub.a and R.sub.b is hydrogen. For example, R.sub.8 of formula
(III) is not hydrogen.
[0278] In some alternative embodiments, N.sub.1 and one of R.sub.a
or R.sub.b of Formula (III) are connected to form a saturated,
partially unsaturated, or unsaturated heterocycle having 5-7 ring
members and the other of R.sub.a or R.sub.b is hydrogen, or absent
as necessary to accommodate ring unsaturation, where the ring is a
5 membered ring, or, for example, the ring is:
##STR00045##
[0279] In some embodiments, at least one of R.sub.2 and R.sub.3 in
the compound of Formula (III) is not hydrogen, or, for example,
R.sub.2 and R.sub.3 are connected to form a saturated carbocycle,
where the saturated carbocycle is cyclopropyl. Alternatively, Z is
N in the compound of Formula (III).
[0280] In some embodiments, the TLR agonist or modulator has the
structure of Formula (IV):
##STR00046##
wherein R.sub.4 is selected from --NR.sub.cR.sub.d and --OR.sub.10;
R.sub.c and R.sub.d are lower alkyl, where the alkyl is optionally
substituted with one or more --OH; R.sub.10 is alkyl, where the
alkyl is optionally substituted with one or more --OH, R.sub.f and
R.sub.g are lower alkyl or R.sub.f and R.sub.g together with the
nitrogen atom to which they are attached form a saturated
heterocyclic ring having 4-6 ring members. For example, R.sub.f and
R.sub.g in the compound of Formula (IV), together with the nitrogen
atom to which they are attached form a saturated heterocyclic ring,
where the heterocyclic ring is pyrrolidine.
[0281] In some alternative embodiments, R.sub.4 of either Formula
(III) or Formula (IV) is --OR.sub.10, where R.sub.10 is alkyl or is
ethyl. In another embodiment, R.sub.4 of either Formula (III) or
Formula (IV) is --NR.sub.cR.sub.d, where both are alkyl or both are
propyl. Moreover, in certain embodiments, at least one of R.sub.c
or R.sub.d is alkyl substituted with one --OH and at least one of
R.sub.c and R.sub.d is
##STR00047##
and the remaining R.sub.c or R.sub.d is propyl.
[0282] In some alternative embodiments, the TLR is a a compound
selected from
##STR00048##
Alternatively, the compound is selected from
##STR00049##
[0283] In some alternative embodiments, the TLR agonist ound is
either
##STR00050##
[0284] In some alternative embodiments, the TLR agonist is a
compound selected from
##STR00051##
[0285] In some alternative embodiments, the TLR agonist is
##STR00052##
[0286] In some alternative embodiments, the TLR agonist is a
compound selected from:
##STR00053## ##STR00054## ##STR00055## ##STR00056##
##STR00057##
[0287] In some embodiments, the immunotherapeutic is a TLR
modulator (e.g., TLR7 and/or TLR8 agonist) that is represented by
structure of Formula (V):
##STR00058##
and metabolites, solvates, tautomers, and prodrugs thereof,
wherein:
[0288] Y is CF.sub.2CF.sub.3, CF.sub.2CF.sub.2R.sup.6 or an aryl or
heteroaryl ring, wherein said aryl and heteroaryl rings are
substituted with one or more groups independently selected from
alkenyl, alkynyl, Br, CN, OH, NR.sup.6R.sup.7, C(.dbd.O)R.sup.8,
NR.sup.6SO.sub.2R.sup.7, (C.sub.1-C.sub.6 alkyl)amino,
R.sup.6OC(.dbd.O)CH.dbd.CH.sub.2--, SR.sup.6 and SO.sub.2R.sup.6,
and wherein the aryl and heteroaryl rings are optionally further
substituted with one or more groups independently selected from F,
Cl, CF.sub.3, CF.sub.3O--, HCF.sub.2O--, alkyl, heteroalkyl and
ArO--;
[0289] R.sup.1, R.sup.3 and R.sup.4 are independently selected from
H, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, cycloalkenyl,
heterocycloalkyl, aryl and heteroaryl, wherein the alkyl, alkenyl,
alkynyl, heteroalkyl, cycloalkyl, cycloalkenyl, heterocycloalkyl,
aryl and heteroaryl are optionally substituted with one or more
groups independently selected from alkyl, alkenyl, alkynyl, F,
Cl.sub.5, Br, I, CN, OR.sup.6, NR.sup.6R.sup.7, C(.dbd.O)R.sup.6,
C(.dbd.O)OR.sup.6, OC(.dbd.O)R.sup.6, C(.dbd.O)NR.sup.6R.sup.7,
(C.sub.1-C.sub.6 alkyl)amino, CH.sub.3OCH.sub.2O--, R.sup.6OC(
O)CH.dbd.CH.sub.2--, NR.sup.6SO.sub.2R.sup.7, SR.sup.6 and
SO.sub.2R.sup.6,
[0290] or R.sup.3 and R.sup.4 together with the atom to which they
are attached form a saturated or partially unsaturated carbocyclic
ring, wherein the carbocyclic ring is optionally substituted with
one or more groups independently selected from alkyl, alkenyl,
alkynyl, F, Cl, Br, I, CN, OR.sup.6, NR.sup.6R.sup.7,
C(.dbd.O)R.sup.6, C(.dbd.O)OR.sup.6, OC(.dbd.O)R.sup.6,
C(.dbd.O)NR.sup.6R.sup.7, (C.sub.1-C.sub.6 alkyl)amino,
CH.sub.3OCH.sub.2O--, R.sup.6OC(.dbd.O)CH.dbd.CH.sub.2--,
NR.sup.6SO.sub.2R.sup.7, SR.sup.6 and SO.sub.2R.sup.6:
[0291] R.sup.2 and R.sup.8 are independently selected from H,
OR.sup.6, NR.sup.6R.sup.7, alkyl, alkenyl, alkynyl, heteroalkyl,
cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl and heteroaryl,
wherein the alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl,
cycloalkenyl, heterocycloalkyl, aryl and heteroaryl are optionally
substituted with one or more groups independently selected from
alkyl, alkenyl, alkynyl, F, Cl, Br.sub.5 I, CN, OR.sup.6,
NR.sup.6R.sup.7, C(.dbd.O)R.sup.6, C(.dbd.O)OR.sup.6,
OC(.dbd.O)R.sup.6, C( O)NR.sup.6R.sup.7, (C.sub.1-C.sub.6
alkyl)amino, CH.sub.3OCH.sub.2O--,
R.sup.6OC(.dbd.O)CH.dbd.CH.sub.2--, NR.sup.6SO.sub.2R.sup.7,
SR.sup.6 and SO.sub.2R.sup.6;
[0292] R.sup.5a, R.sup.5b, and R.sup.5c are independently H, F, Cl,
Br, I.sub.5 OMe.sub.5 CH.sub.3, CH.sub.2F.sub.5 CHF.sub.2 or
CF.sub.3; and
[0293] R.sup.6 and R.sup.7 are independently selected from H.sub.5
alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, cycloalkenyl,
heterocycloalkyl, aryl and heteroaryl, wherein said alkyl, alkenyl,
alkynyl, heteroalkyl, cycloalkyl, cycloalkenyl, heterocycloalkyl,
aryl and heteroaryl are optionally substituted with one or more
groups independently selected from alkyl, alkenyl, alkynyl, F, Cl,
Br, I, CN, OR.sup.6, NR.sup.6R.sup.7, C(.dbd.O)R.sup.6,
C(.dbd.O)OR.sup.6, OC(O)R.sup.6, C(O)NR.sup.6R.sup.7,
(C.sub.1-C.sub.6 alkyl)amino, CH.sub.3OCH.sub.2O--, R.sup.6OC(
O)CH.dbd.CH.sub.2--, NR.sup.6SO.sub.2R.sup.7, SR.sup.6 and
SO.sub.2R.sup.6,
[0294] or R.sup.6 and R.sup.7 together with the atom to which they
are attached form a saturated or partially unsaturated heterocyclic
ring, wherein said heterocyclic ring is optionally substituted with
one or more groups independently selected from alkyl, alkenyl,
alkynyl, F, Cl, Br, I, CN, OR.sup.6, NR.sup.6R.sup.7,
C(.dbd.O)R.sup.6, C(.dbd.O)OR.sup.6, OC(.dbd.O)R.sup.6,
C(.dbd.O)NR.sup.6R.sup.7, (C.sub.1-C.sub.6 alkyl)amino,
CH.sub.3OCH.sub.2O--, R.sup.6OC(.dbd.O)CH.dbd.CH.sub.2--,
NR.sup.6SO.sub.2R.sup.7, SR.sup.6 and SO.sub.2R.sup.6. In certain
embodiments, R.sup.1, R.sup.3 and R.sup.4 are each hydrogen. In
certain embodiments, R.sup.5a, R.sup.5b and R.sup.5c are each
hydrogen. WO 2007024612 A2, the disclosure of which is incorporated
by reference in its entirety.
[0295] In some embodiments of the compound of Formula (V), R.sup.2
is OR.sup.6. In some embodiments, R.sup.6 is alkyl, such as
(1-4C)alkyl. In particular embodiments, R.sup.6 is ethyl.
[0296] In some embodiments of the compound of Formula (V), R.sup.2
is NR.sup.6R.sup.7. In some embodiments, R.sup.6 and R.sup.7 are
independently H, alkyl, such as (1-6C)alkyl, or heteroalkyl, such
as (1-4C)alkoxy(2-4C)alkyl. In particular embodiments, R.sup.6 and
R.sup.7 are independently H, ethyl, propyl, or
CH.sub.2CH.sub.2OCH.sub.3. In some embodiments of the compound of
Formula V, Y is aryl, such as phenyl. In some embodiments, the aryl
is substituted with C(.dbd.O)R.sup.8, such as in
para-R.sup.8C(.dbd.O)phenyl. In some embodiments, R.sup.8 is
OR.sup.6, NR.sup.6R.sup.7 or heterocycloalkyl. In some embodiments,
R.sup.6 and R.sup.7 are independently H or alkyl, such as
(1-6C)alkyl. In some other embodiments, R.sup.6 and R.sup.7
together with the nitrogen atom to which they are attached form a
4-6 membered azacycloalkyl ring, such as pyrrolidinyl. In some
embodiments, Y is
##STR00059##
[0297] In some embodiments of the compound of Formula (V), Y is
CF.sub.2CF.sub.3.
[0298] In some embodiments, the immunotherapeutic is a TLR
modulator (e.g., TLR8 agonist) that is represented by structure of
formula (VI):
##STR00060##
and metabolites, solvates, tautomers, and pharmaceutically
acceptable prodrugs and salts thereof, wherein:
[0299] Z is H, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl,
heterocycloalkyl, aryl, heteroaryl. OR.sup.6 or NR.sup.6R.sup.7,
wherein said alkyl, alkenyl, alkynyl; heteroalkyl, cycloalkyl,
heterocycloalkyl, aryl and heteroaryl are optionally substituted
with one or more groups independently selected from alkyl, alkenyl,
alkynyl, F, Cl.sub.3 Br, I, CN, OR.sup.6, NR.sup.6R.sup.7,
C(.dbd.O)R.sup.6, C(.dbd.O)OR.sup.6, OC(.dbd.O)R.sup.6,
C(.dbd.O)NR.sup.6R.sup.7, CC.sub.1-C.sub.6alkyl)amino,
CH.sub.3OCH.sub.2O--, R.sup.6OCC.dbd.O)CH.dbd.CH.sub.2--,
NR.sup.6SO.sub.2R.sup.7, SR.sup.6 and SO.sub.2R.sup.6;
[0300] R.sup.1, R.sup.2, R.sup.3 and R.sup.4 are independently
selected from H, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl,
cycloalkenyl, heterocycloalkyl, aryl and heteroaryl, wherein said
alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, cycloalkenyl,
heterocycloalkyl, aryl, and heteroaryl are optionally substituted
with one or more groups independently selected from alkyl, alkenyl,
alkynyl, F, Cl, Br, I, CN, OR.sup.6, NR.sup.6R.sup.7,
CC.dbd.O)R.sup.6, C(.dbd.O)OR.sup.6, OC(.dbd.O)R.sup.6,
CC.dbd.O)NR.sup.6R.sup.7, (C.sub.1-C.sub.6 alkyl)amino,
CH.sub.3OCH.sub.2O--, R.sup.6OCC.dbd.O)CH.dbd.CH.sub.2--,
NR.sup.6SO.sub.2R.sup.7, SR.sup.6 and SO.sub.2R.sup.6,
[0301] or R.sup.1 and R.sup.2 together with the atom to which they
are attached form a saturated or partially unsaturated carbocyclic
ring, wherein said carbocyclic ring is optionally substituted with
one or more groups independently selected from alkyl, alkenyl,
alkynyl, F, Cl, Br, I, CN, OR.sup.6, NR.sup.6R.sup.7,
C(.dbd.O)R.sup.6, CC.dbd.O)OR.sup.6, OC(.dbd.O)R.sup.6,
CC.dbd.O)NR.sup.6R.sup.7, (C.sub.1-C.sub.6 alkyl)amino,
CH.sub.3OCH.sub.2O--, R.sup.6OC(.dbd.O)CH.dbd.CH.sub.2--,
NR.sup.6SO.sub.2R.sup.7, SR.sup.6 and SO.sub.2R.sup.6;
[0302] or R.sup.3 and R.sup.4 together are oxo;
[0303] each R.sup.5 is independently selected from H, F, Cl, Br, I,
OMe, CH.sub.3, CH.sub.2F, CHF.sub.2, CF.sub.3 and
CF.sub.2CF.sub.3;
[0304] R.sup.6 and R.sup.7 are independently selected from H,
alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, cycloalkenyl,
heterocycloalkyl, aryl, and heteroaryl, wherein said alkyl,
alkenyl, alkynyl, heteroalkyl, cycloalkyl, cycloalkenyl,
heterocycloalkyl, aryl, and heteroaryl are optionally substituted
with one or more groups independently selected from alkyl, alkenyl,
alkynyl, F, Cl, Br, I, CN, OR.sup.6, NR.sup.6R.sup.7,
CC.dbd.O)R.sup.6, C(=0)0R.sup.6, 0C(=0)R.sup.6,
CC.dbd.O)NR.sup.6R.sup.7, (C.sub.1-C.sub.6 alkyl)amino,
CH.sub.3OCH.sub.2O--, R.sup.6OC(.dbd.O)CH.dbd.CH.sub.2--,
NR.sup.6SO.sub.2R.sup.7, SR.sup.6 and SO.sub.2R.sup.6;
[0305] or R.sup.6 and R.sup.7 together with the atom to which they
are attached form a saturated or partially unsaturated heterocyclic
ring, wherein the heterocyclic ring is optionally substituted with
one or more groups independently selected from alkyl, alkenyl,
alkynyl, F, Cl, Br, I, CN, OR.sup.6, NR.sup.6R.sup.7,
CC.dbd.O)R.sup.6, C(=0)0R.sup.6, 0C(=0)R.sup.6,
C(.dbd.O)NR.sup.6R.sup.7, (C.sub.1-C.sub.6 alkyl)amino,
CH.sub.3OCH.sub.2O--, R.sup.6OC(.dbd.O)CH.dbd.CH.sub.2--,
NR.sup.6SO.sub.2R.sup.7, SR.sup.6 and SO.sub.2R.sup.6; and n is O,
1, 2, 3 or 4. WO2007040840A2, the disclosure of which is
incorporated by reference in its entirety.
[0306] In some embodiments, the immunotherapeutic is a TLR
modulator (e.g., TLR8 agonist) that is represented by structure of
Formula (VI):
##STR00061##
and metabolites, solvates, tautomers, and pharmaceutically
acceptable salts and prodrugs thereof, wherein.
[0307] Z is H, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl,
heterocycloalkyl, aryl, heteroaryl, OR.sup.6 or NR.sup.6R.sup.7,
wherein the alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl,
heterocycloalkyl, aryl and heteroaryl are optionally substituted
with one or more groups independently selected from alkyl, alkenyl,
alkynyl, F, Cl, Br, I, CN, OR.sup.6, NR.sup.6R.sup.7,
C(.dbd.O)R.sup.6, C(.dbd.O)OR.sup.6, OC(.dbd.O)R.sup.6,
C(.dbd.O)NR.sup.6R.sup.7, (C.sub.1-C.sub.6 alkyl)amino,
CH.sub.3OCH.sub.2O--, R.sup.6OCC.dbd.O)CH.dbd.CH.sub.2,
NR.sup.6SO.sub.2R.sup.7, SR.sup.6 and SO.sub.2R.sup.6;
[0308] R.sup.1, R.sup.2, R.sup.3 and R.sup.4 are independently
selected from H, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl,
cycloalkenyl, heterocycloalkyl, aryl and heteroaryl, wherein said
alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, cycloalkenyl,
heterocycloalkyl, aryl, and heteroaryl are optionally substituted
with one or more groups independently selected from alkyl, alkenyl,
alkynyl, F, Cl, Br, I.sub.9 CN, OR.sup.6, NR.sup.6R.sup.7,
C(.dbd.O)R.sup.6, C(.dbd.O)OR.sub.6, OC(.dbd.O)R.sup.6,
C(.dbd.O)NR.sup.6R.sup.7, (C.sub.1-C.sub.6 alkyl)amino,
CH.sub.3OCH.sub.2O--, R.sup.6OCC.dbd.O)CH.dbd.CH.sub.2--,
NR.sup.6SO.sub.2R.sup.7, SR.sup.6 and SO.sub.2R.sup.6,
[0309] or R.sup.1 and R.sup.2 together with the atom to which they
are attached form a saturated or partially unsaturated carbocyclic
ring, wherein said carbocyclic ring is optionally substituted with
one or more groups independently selected from alkyl, alkenyl,
alkynyl, F, Cl, Br, I, CN, OR.sup.6, NR.sup.6R.sup.7,
C(.dbd.O)R.sup.6, C(.dbd.O)OR.sup.6, OC(.dbd.O)R.sup.6,
C(.dbd.O)NR.sup.6R.sup.7, (C.sub.1-C.sub.6alkyl)amino,
CH.sub.3OCH.sub.2O--, R.sup.6OC(.dbd.O)CH.dbd.CH.sub.2--,
NR.sup.6SO.sub.2R.sup.7, SR.sup.6 and SO.sub.2R.sup.6,
[0310] or R.sup.3 and R.sup.4 together are oxo;
[0311] R.sup.5 is H, F, Cl, Br, I, OMe, CH.sub.3, CH.sub.2F,
CHF.sub.2, CF.sub.3 or CF.sub.2CF.sub.3;
[0312] R.sup.6 and R.sup.7 are independently selected from H,
alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, cycloalkenyl,
heterocycloalkyl, aryl, and heteroaryl, wherein said alkyl,
alkenyl, alkynyl, heteroalkyl, cycloalkyl, cycloalkenyl,
heterocycloalkyl, aryl, and heteroaryl are optionally substituted
with one or more groups independently selected from alkyl, alkenyl,
alkynyl, F, Cl, Br, I, CN, OR.sup.6, NR.sup.6R.sub.7,
C(.dbd.O)R.sup.6, C(=0)0R.sup.6, 0C(=0)R.sup.6,
C(O)NR.sup.6R.sup.7, (C.sub.1-C.sub.6 alkyl)amino.sub.5
CH.sub.3OCH.sub.2O--, R.sup.6OC(.dbd.O)CH.dbd.CH.sub.2--,
NR.sup.6SO.sub.2R.sup.7, SR.sup.6 and SO.sub.2R.sup.6;
[0313] or R.sup.6 and R.sup.7 together with the atom to which they
are attached form a saturated or partially unsaturated heterocyclic
ring, wherein said heterocyclic ring is optionally substituted with
one or more groups independently selected from alkyl, alkenyl,
alkynyl, F, Cl, Br, I, CN, OR.sup.6, NR.sup.6R.sup.7,
C(.dbd.O)R.sup.6, C(.dbd.O)OR.sup.6, OC(.dbd.O)R.sup.6,
C(.dbd.O)NR.sup.6R.sup.7, (C.sub.1-C.sub.6 alkyl)amino,
CH.sub.3OCH.sub.2O--, R.sup.6OC(.dbd.O)CH.dbd.CH.sub.2--,
NR.sup.6SO.sub.2R.sup.7, SR.sup.6 and SO.sub.2R.sup.6, and
[0314] n is O, 1, 2, 3 or 4.
[0315] In some embodiments, Z is OR.sup.6. In some embodiments,
R.sup.6 is alkyl, such as (1-6C)alkyl. In particular embodiments,
R.sup.6 is ethyl, propyl, isopropyl or isobutyl.
[0316] In some embodiments, Z is NR.sup.6R.sup.7. In some
embodiments, R.sup.6 and R.sup.7 are independently H or alkyl, such
as (1-6C)alkyl. In some embodiments, R.sup.6 and R.sup.7 are ethyl.
In some embodiments, n is O or 1.
[0317] In some embodiments, R.sup.5 is CF.sub.2CF.sub.3. In certain
embodiments, R.sup.3 is H or alkyl, such as (1-4C)alkyl, and
R.sup.4 is H. In certain embodiments, R is alkyl, such as
(1-4C)alkyl. In some embodiments, R is methyl. In other particular
embodiments, R.sup.3 is H. In some embodiments, R is H or alkyl,
such as (1-4C)alkyl and R is H. In some embodiments, R.sup.1 is
alkyl. In some embodiments, R.sup.1 is methyl. In some particular
embodiments, R.sup.1 is H.
[0318] In some embodiments, the activating moiety is a TLR7 and/or
TLR8 agonist that is represented by structure of Formula (XV):
##STR00062##
wherein ring A represents a 6-10 membered aromatic carbocyclic ring
or a 5-10 membered heteroaromatic ring; R represents a halogen
atom, an alkyl group, a hydroxyalkyl group, a haloalkyl group, an
alkoxy group, a hydroxyalkoxy group, a haloalkoxy group, amino
group, an alkylamino group, a dialkylamino group, or a 4-7 membered
cyclic group containing in the ring 1-2 hetero atoms selected from
1-2 nitrogen atoms and optionally 0-1 oxygen atom or 0-1 sulfur
atom; n represents an integer of 0-2, and when n is 2, the Rs may
be the same or different; Z.sup.1 represents a substituted or
unsubstituted alkylene group or a substituted or unsubstituted
cycloalkylene group; X.sup.2 represents oxygen atom, sulfur atom,
SO.sub.2, NR.sup.5, CO, CONR.sup.5, NR.sup.5CO, SO.sub.2NR.sup.5,
NR.sup.5SO.sub.2, NR.sup.5CONR.sup.6 or NR.sup.5CSNR.sup.6 (in
which R.sup.5 and R.sup.6 are each independently hydrogen atom, a
substituted or unsubstituted alkyl group, or a substituted or
unsubstituted cycloalkyl group); Y.sup.1, Y.sup.2 and Y.sup.3
represent each independently a single bond or an alkylene group;
X.sup.1 represents oxygen atom, sulfur atom, SO.sub.2, NR.sup.4
(wherein R.sup.4 is hydrogen atom or an alkyl group) or a single
bond; R.sup.2 represents hydrogen atom, a substituted or
unsubstituted alkyl group, a substituted or unsubstituted alkenyl
group, a substituted or unsubstituted alkynyl group or a
substituted or unsubstituted cycloalkyl group; and R.sup.1
represents hydrogen atom, hydroxy group, an alkoxy group, an
alkoxycarbonyl group, a haloalkyl group, a haloalkoxy group, a
substituted or unsubstituted aryl group, a substituted or
unsubstituted heteroaryl group or a substituted or unsubstituted
cycloalkyl group. The linker is linked to one of the possible
linking site of the angonist, such as to --NH.sub.2.
[0319] In some embodiments, R.sup.1 represents hydrogen, hydroxyl,
or a C.sub.1-C.sub.6 alkoxy, C.sub.2-C.sub.5alkoxycarbonyl,
C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6 haloalkoxy,
C.sub.6-C.sub.10 aryl, C.sub.5-C.sub.10heteroaryl or
C.sub.1-C.sub.8 cycloalkyl group, each group being optionally
substituted by one or more substituents independently selected from
halogen, hydroxyl, a C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6
haloalkyl, C.sub.1-C.sub.6 alkoxy, C.sub.1-C.sub.6haloalkoxy,
C.sub.2-C.sub.5 alkoxycarbonyl, amino (NH.sub.2),
(mono)-C.sub.1-C.sub.6 alkylamino and (di)-C.sub.1-C.sub.6
alkylamino group;
Y.sup.1 represents a single bond or C.sub.1-C.sub.6 alkylene;
X.sup.1 represents a single bond, an oxygen, sulphur atom,
sulphonyl (SO.sub.2) or NR.sup.3; Z.sup.1 represents a
C.sub.2-C.sub.6 alkylene or C.sub.3-C.sub.8 cycloalkylene group,
each group being optionally substituted by at least one hydroxyl;
X.sup.2 represents NR.sup.4; Y.sup.2 represents a single bond or
C.sub.1-C.sub.6 alkylene; Y.sup.3 represents a single bond or
C.sub.1-C.sub.6 alkylene; n is an integer 0, 1 or 2; R represents
halogen or a C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 hydroxyalkyl,
C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6alkoxy, C.sub.1-C.sub.6
hydroxyalkoxy, C.sub.1-C.sub.6 haloalkoxy, amino (NH.sub.2),
(mono)-C.sub.1-C.sub.6 alkylamino, (di)-C.sub.1-C.sub.6 alkylamino
group or a C.sub.3-C.sub.8saturated heterocyclic ring containing a
ring nitrogen atom and optionally one or more further heteroatoms
independently selected from nitrogen, oxygen and sulphur, the
heterocyclic ring being optionally substituted by one or more
substituents independently selected from halogen, hydroxyl, oxo,
C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, C.sub.2-C.sub.5
alkylcarbonyl and C.sub.2-C.sub.5alkoxycarbonyl; R.sup.2 represents
hydrogen or a C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl or C.sub.3-C.sub.8 cycloalkyl group, each
group being optionally substituted by one or more substituents
independently selected from halogen, hydroxyl or a C.sub.1-C.sub.6
alkoxy, a C.sub.2-C.sub.10 acyloxy, group selected from a C.sub.2-5
alkylcarbonyloxy group, a C.sub.2-C.sub.5 alkenylcarbonyloxy group,
a C.sub.2-C.sub.5 alkynylcarbonyloxy group, a C.sub.6-C.sub.9
arylcarbonyloxy group and a C.sub.5-C.sub.9heteroarylcarbonyloxy
group, each of which acyloxy groups may be optionally substituted
by one or more substituents independently selected from halogen,
hydroxyl, C.sub.1-C.sub.3 alkoxy and phenyl providing that the
total number of carbon atoms in the acyloxy group does not exceed
10, amino (NH.sub.2), (mono)-C.sub.1-C.sub.6 alkylamino,
(di)-C.sub.1-C.sub.6 alkylamino group and a C.sub.3-C.sub.8
saturated heterocyclic ring containing a ring nitrogen atom and
optionally one or more further heteroatoms independently selected
from nitrogen, oxygen and sulphur, the heterocyclic ring in turn
being optionally substituted by one or more substituents
independently selected from halogen, hydroxyl, oxo, C.sub.1-C.sub.6
alkyl, C.sub.1-C.sub.6 alkoxy, C.sub.2-C.sub.5alkylcarbonyl and
C.sub.2-C.sub.5 alkoxycarbonyl group; R.sup.3 represents hydrogen
or C.sub.1-C.sub.6 alkyl; R.sup.4 represents CO.sub.2R.sup.5,
SO.sub.2R.sup.5, COR.sup.5, SO.sub.2NR.sup.6R.sup.7 and
CONR.sup.6R.sup.7; R.sup.5 independently represents (i) 3- to
8-membered heterocyclic ring containing 1 or 2 heteroatoms selected
from a ring group NR.sup.8, S(O).sub.m or oxygen, the 3- to
8-membered heterocyclic ring being optionally substituted by one or
more substituents independently selected from halogen, hydroxyl or
a C.sub.1-C.sub.6 alkyl and C.sub.1-C.sub.6 alkoxy group, or (ii) a
C.sub.6-C.sub.10 aryl or C.sub.5-C.sub.10 heteroaryl group, each of
which may be optionally substituted by one or more substituents
independently selected from halogen, cyano, C.sub.1-C.sub.6 alkyl,
C.sub.1-C.sub.3 haloalkyl, carboxyl, S(O).sub.mR.sup.9, OR.sup.10,
CO.sub.2R.sup.10, SO.sub.2NR.sup.10R.sup.11, CONR.sup.10R.sup.11,
NR.sup.10R.sup.11, NR.sup.10SO.sub.2R.sup.9,
NR.sup.10CO.sub.2R.sup.9, NR.sup.10COR.sup.9, or (iii) a
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl or C.sub.3-C.sub.8 cycloalkyl group, each of which may be
optionally substituted by one or more substituents independently
selected from halogen, CN, C.sub.3-C.sub.8cycloalkyl,
S(O).sub.pR.sup.12, OR.sup.13, COR.sup.13, CO.sub.2R.sup.13,
SO.sub.2NR.sup.13R.sup.14, CONR.sup.13R.sup.14, NR.sup.13R.sup.14,
NR.sup.13SO.sub.2R.sup.12, NR.sup.13CO.sub.2R.sup.12,
NR.sup.13COR.sup.12, NR.sup.13SO.sub.2R.sup.12 or a
C.sub.6-C.sub.10 aryl or C.sub.5-C.sub.10 heteroaryl group or a
heterocyclic ring, the latter three groups may be optionally
substituted by one or more substituents independently selected from
C.sub.1-C.sub.6 alkyl (optionally substituted by hydroxy,
C.sub.1-C.sub.6 alkoxy, C.sub.1-C.sub.6 alkoxycarbonyl, amino,
C.sub.1-C.sub.6 alkylamino, di-C.sub.1-C.sub.6 alkylamino,
NH.sub.2C(O)--, C.sub.1-C.sub.6 alkylNHC(O), di-C.sub.1-C.sub.6
alkyl NC(O), --OCH.sub.2CH.sub.2OH, pyrrolidinyl,
pyrrolidinylcarbonyl, furanyl, piperidyl, methylpiperidyl or
phenyl), C.sub.2-C.sub.6 alkenyl (optionally substituted by
phenyl), halogen, hydroxy, cyano, carboxy, amino, C.sub.1-C.sub.6
alkylamino, di-C.sub.1-C.sub.6 alkylamino, NH.sub.2C(O)--,
C.sub.1-C.sub.6 alkyl NHC(O)--, di-C.sub.1-C.sub.6 alkyl NC(O),
C.sub.1-C.sub.6 alkoxycarbonyl, C.sub.1-C.sub.6 alkylsulphonyl,
C.sub.1-C.sub.6 alkylcarbonylamino, C.sub.1-C.sub.6
alkylcarbonylmethylamino, phenyl (optionally substituted by
hydroxy, fluoro or methyl), pyrrolidinyl, pyridyl, piperidinyl,
benzothiazolyl or pyrimidinyl; R.sup.6 represents hydrogen or a
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, C.sub.3-C.sub.8 cycloalkyl group or heterocyclic ring,
each of which may be optionally substituted by one or more
substituents independently-selected from halogen, hydroxyl, oxo,
cyano, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl, C.sub.3-C.sub.8 cycloalkyl, OR.sup.15,
S(O).sub.qR.sup.15, CO.sub.2R.sup.16, COR.sup.16,
NR.sup.16R.sup.17, CONR.sup.16R.sup.17, NR.sup.16COR.sup.17,
NR.sup.16CO.sub.2R.sup.15, SO.sub.2NR.sup.16R.sup.17,
NR.sup.16SO.sub.2R.sup.15, or a C.sub.6-C.sub.10 aryl or
C.sub.5-C.sub.10 heteroaryl group or heterocyclic ring, the latter
three groups being optionally substituted by one or more
substituents independently selected from, C.sub.1-C.sub.6 alkyl,
C.sub.3-C.sub.8 cycloalkyl, halogen, S(O).sub.qR.sup.15,
CO.sub.2R.sup.16, COR.sup.16, hydroxy or cyano; and R.sup.7
represents hydrogen, a C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, or C.sub.3-C.sub.8 cycloalkyl
group, each group may be optionally substituted by one or more
substituents independently selected from halogen,
C.sub.3-C.sub.8cycloalkyl, a C.sub.6-C.sub.10 aryl or
C.sub.5-C.sub.10 heteroaryl group, carboxy, cyano, OR.sup.15,
hydroxy or NR.sup.18R.sup.19, or R.sup.6 and R.sup.7 together with
the nitrogen atom to which they are attached fowl a 3- to
8-membered saturated or partially saturated heterocyclic ring,
optionally containing further heteroatoms or heterogroups selected
from nitrogen. S(O).sub.m or oxygen, the heterocyclic ring, may be
optionally substituted by one or more substituents independently
selected from halogen, hydroxyl, carboxyl, cyano, OR.sup.20,
NR.sup.21R.sup.22, S(O).sub.qR.sup.23, COR.sup.24,
CO.sub.2R.sup.24, NR.sup.24R.sup.25, CONR.sup.24R.sup.25,
NR.sup.24COR.sup.25, NR.sup.24CO.sub.2R.sup.23,
SO.sub.2NR.sup.24R.sup.25, NR.sup.24SO.sub.2R.sup.23,
C.sub.6-C.sub.10 aryl, C.sub.5-C.sub.10 heteroaryl group,
heterocyclic ring, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl or C.sub.3-C.sub.8cycloalkyl group, the
latter seven groups being optionally substituted by one or more
substituents independently selected from halogen, hydroxyl, oxo,
cyano, OR.sup.20, S(O).sub.qR.sup.23, COR.sup.24, CO.sub.2R.sup.24,
NR.sup.24R.sup.25, CONR.sup.24R.sup.25, NR.sup.24CO.sub.2R.sup.23,
NR.sup.24COR.sup.25, SO.sub.2NR.sup.24R.sup.25,
NR.sup.24SO.sub.2R.sup.23, a heterocyclic ring or a
C.sub.6-C.sub.10 aryl or C.sub.5-C.sub.10 heteroaryl group, the
latter three groups being optionally substituted by one or more
substituents independently selected from C.sub.1-C.sub.6 alkyl,
halogen, hydroxy or cyano; R.sup.8 represents hydrogen,
CO.sub.2R.sup.26, COR.sup.26, SO.sub.2R.sup.26, C.sub.1-C.sub.6
alkyl or C.sub.3-C.sub.6 cycloalkyl group, each group may be
optionally substituted by one or more substituents independently
selected from halogen, hydroxyl, and NR.sup.27R.sup.28; R.sup.10,
R.sup.11, R.sup.16, R.sup.17, R.sup.18, R.sup.19, R.sup.21,
R.sup.22, R.sup.26, R.sup.27 or R.sup.28 each independently
represents hydrogen, and a C.sub.1-C.sub.6 alkyl or C.sub.3-C.sub.6
cycloalkyl group; R.sup.24 and R.sup.25 each independently
represents hydrogen, and a C.sub.1-C.sub.6 alkyl or C.sub.3-C.sub.6
cycloalkyl group; or R.sup.24 and R.sup.25 together with the
nitrogen atom to which they are attached form a 3- to 8-membered
saturated or partially saturated heterocyclic ring, optionally
containing further heteroatoms or heterogroups selected from
nitrogen, S(O).sub.m or oxygen; R.sup.9, R.sup.12, R.sup.15 and
R.sup.23 represent C.sub.1-C.sub.6 alkyl or C.sub.3-C.sub.6
cycloalkyl; R.sup.13 and R.sup.14 are defined as for R.sup.6 and
R.sup.7 respectively; R.sup.20 represents a C.sub.1-C.sub.6 alkyl
optionally substituted by one or more substituents independently
selected from halogen, hydroxyl or OR.sup.23; m, p, q and r each
independently represent an integer 0, 1 or 2; and A represents a
C.sub.6-C.sub.10 aryl or C.sub.5-C.sub.12 heteroaryl group. See
WO2008004948A1, U.S. Pat. No. 8,138,172, and U.S. Pat. No.
8,575,180 the disclosure of which is incorporated by reference.
[0320] In some embodiments, the activating moiety is a TLR7 and/or
TLR8 agonist having the structure of:
##STR00063##
wherein R is Me or H.
[0321] In some embodiments, the activating moiety is a TLR7 and/or
TLR8 agonist having the structure of:
##STR00064## ##STR00065##
[0322] In some embodiments, the activating moiety is a TLR7 and/or
TLR8 agonist having the structure of Formula (XVI):
##STR00066##
wherein: R.sup.1 is independently H, --C(O)R.sup.3, or a racemic,
L-, or D-amino acid group --C(O)CHNH.sub.2R.sup.4, wherein R.sup.3
is a substituted or unsubstituted alkyl, and R.sup.4 is H, or a
substituted or unsubstituted alkyl; R.sup.2 is H, O, OR.sup.5, or
N(R.sup.6).sub.2, wherein R.sup.5 is independently H or alkyl, and
wherein R.sup.6 is independently H, substituted or unsubstituted
alkyl, cycloalkyl, or together with nitrogen forms a substituted or
unsubstituted heterocycloalkyl ring; and wherein if R is --OH, at
least one of the R groups is a racemic, L-, or D-amino acid group
--C(O)CHNH.sub.2R.sup.4. See U.S. Pat. No. 6,924,271, the
disclosure of which is incorporated by reference in its
entirety.
[0323] In some embodiments, at least one of the R.sup.1 groups is a
racemic. L-, or D-amino acid group --C(O)CHNH.sub.2R.sup.4, wherein
R.sup.4 is a substituted or unsubstituted alkyl, and wherein the
remaining R.sup.1 groups are H; R.sup.2 is OR.sup.5 or
N(R.sup.6).sub.2, wherein R.sup.5 is independently selected from H
or alkyl, and wherein R is independently H, substituted or
unsubstituted alkyl, cycloalkyl, or together with nitrogen forms a
substituted or unsubstituted heterocycloalkyl ring.
[0324] In some embodiments, at least one of the R.sup.1 groups is a
L-amino acid group --C(O)CHNH.sub.2R.sup.4, wherein R.sup.4 is a
substituted or unsubstituted alkyl, and wherein the remaining
R.sup.1 groups are H; R.sup.2 is OR.sup.5 or N(R.sup.6).sub.2,
wherein R.sup.4 is a substituted alkyl, and wherein R.sup.6 is
independently H or substituted or unsubstituted alkyl.
[0325] In some embodiments, at least one of the R.sup.1 groups is a
L-amino acid group --C(O)CHNH.sub.2R, wherein R.sup.4 is
--CH(CH.sub.3).sub.2, and wherein the remaining R.sup.1 groups are
H; and R.sup.2 is OH.
[0326] In some embodiments, the TLR7 and/or agonist is selected
from the group consisting of:
##STR00067## ##STR00068## ##STR00069## ##STR00070##
[0327] In some embodiments, the activating moiety is a TLR7 and/or
TLR8 agonist having the structure of:
##STR00071## ##STR00072##
wherein:
[0328] each R.sup.1 is H, or a substituted or unsubstituted alkyl,
alkenyl, or alkynyl, which may be interrupted by one or more O, S,
or N heteroatoms, or a substituted or unsubstituted aryl or
heteroaryl;
[0329] R.sup.2 is H, OH, SH, halo, or a substituted or
unsubstituted alkyl, alkenyl, or alkynyl, which may be interrupted
by one or more O, S, or N heteroatoms, or a substituted or
unsubstituted --O-(alkyl), --O-(aryl), --O-(heteroaryl),
--S-(alkyl), --S-(aryl), --S-(heteroaryl), aryl, or heteroaryl;
[0330] R.sup.3 is H, OH, or SH, or a substituted or unsubstituted
alkyl, alkenyl, alkynyl, aryl, heteroaryl, --O-(alkyl), --O-(aryl),
--O-(heteroaryl), --S-(alkyl), --S-(aryl), --S-(heteroaryl),
--NH(alkyl), --NH(aryl), --NH(heteroaryl), --NH(R.sup.4)(alkyl),
--NH(R.sup.4)(aryl), or --NH(R.sup.4)(heteroaryl), wherein R.sup.4
is a substituted or unsubstituted alkyl;
[0331] X is O or S;
[0332] V is H, halo, OH, OR.sup.4, SH, SR.sup.4, or a substituted
or unsubstituted alkyl or aryl; and
[0333] Z is H, halo, OH, OR.sup.4, SH, or SR.sup.4. See U.S. Pat.
No. 7,576,068, the disclosure of which is incorporated by reference
in its entirety.
[0334] In some embodiments, the activating moiety is a TLR7 and/or
TLR8 agonist having the structure of Formula (XVIII):
##STR00073##
wherein: Y--Z is --CR.sup.4R.sup.5--,
--CR.sup.4R.sup.5--CR.sup.4R.sup.5--, --C(O)CR.sup.4R.sup.5--,
--CR.sup.4R.sup.5C(O)--, --NR.sup.8C(O)--, --C(O)NR.sup.8--,
CR.sup.4R.sup.5S(O).sub.2--, or --CR.sup.5.dbd.CR.sup.5--; L.sup.1
is --NR.sup.8--, --O--, --S--, --N(R.sup.8)C(O)--, --S(O).sub.2--,
--S(O)--C(O)N(R.sup.8)--, --N(R.sup.8)S(O).sub.2--,
--S(O).sub.2N(R.sup.8)-- or a covalent bond: R.sup.1 is alkyl,
substituted alkyl, haloalkyl, alkenyl, substituted alkenyl,
alkynyl, substituted alkynyl, heteroalkyl, substituted heteroalkyl,
carbocyclyl, substituted carbocyclyl, carbocyclylalkyl, substituted
carbocyclylalkyl, heterocyclyl, substituted heterocyclyl,
heterocyclylalkyl, or substituted heterocyclylalkyl, arylalkyl,
substituted arylalkyl, heteroarylalkyl, substituted
heteroarylalkyl, carbocyclylheteroalkyl, substituted
carbocyclylheteroalkyl, heterocyclylheteroalkyl, substituted
heterocyclylheteroalkyl, arylheteroalkyl, substituted
arylheteroaryl, heteroarylheteroalkyl, or substituted
heteroarylheteroalkyl; X.sup.1 is alkylene, substituted alkylene,
heteroalkylene, substituted heteroalkylene, alkenylene, substituted
alkenylene, alkynylene, substituted alkynylene, carbocyclylene,
substituted carbocyclylene, heterocyclylene, substituted
heterocyclylene, --NR.sup.8--, --O--, --C(O)--, --S(O)--,
--S(O).sub.2--, or a bond; D is carbocyclyl, substituted
carbocyclyl, heterocyclyl or substituted heterocyclyl wherein said
carbocyclyl, substituted carbocyclyl, heterocyclyl or substituted
heterocyclyl is substituted with one or two
-L.sup.2-NR.sup.6R.sup.7; or D is a heterocyclyl, substituted
heterocyclyl, heteroaryl or substituted heteroaryl wherein said
heterocyclyl, substituted heterocyclyl, heteroaryl or substituted
heteroaryl comprises one to four nitrogen atoms; each L.sup.2 is
independently alkylene, substituted alkylene, heteroalkylene,
substituted heteroalkylene, or a covalent bond; each R.sup.3 is
independently halogen, cyano, azido, nitro, alkyl, substituted
alkyl, hydroxyl, amino, heteroalkyl, substituted heteroalkyl,
alkoxy, haloalkyl, haloalkoxy, --CHO, --C(O)OR.sup.8,
--S(O)R.sup.8, --S(O).sub.2R.sup.8; --C(O)NR.sup.9R.sup.10,
--N(R.sup.9)C(O)R.sup.8, carbocyclyl, substituted carbocyclyl,
carbocyclylalkyl, substituted carbocyclylalkyl, alkenyl,
substituted alkenyl, alkynyl, substituted alkynyl,
--S(O).sub.2NR.sup.9R.sup.10, --N(R.sup.9)S(O).sub.2R.sup.8,
--N(R.sup.9)S(O).sub.2OR.sup.10, --OS(O).sub.2NR.sup.9R.sup.10; n
is 0, 1, 2, 3, 4 or 5; R.sup.4 and R.sup.5 are each independently
H, alkyl, substituted alkyl, haloalkyl, heteroalkyl, substituted
heteroalkyl, carbocyclyl, substituted carbocyclyl,
carbocyclylalkyl, substituted carbocyclylalkyl, heterocyclyl,
substituted heterocyclyl, heterocyclylalkyl, substituted
heterocyclylalkyl, arylalkyl, substituted arylalkyl,
heteroarylalkyl, substituted heteroarylalkyl,
carbocyclylheteroalkyl, substituted carbocyclylheteroalkyl,
heterocyclylheteroalkyl, substituted heterocyclylheteroalkyl,
arylheteroalkyl, substituted arylheteroalkyl,
heteroarylheteroalkyl, or substituted heteroarylheteroalkyl, cyano,
azido, OR.sup.8, --C(O)H, --C(O)R.sup.8, --S(O)R.sup.8,
--S(O).sub.2R.sup.8, --C(O)OR.sup.8, or --C(O)NR.sup.9R.sup.10; or
R.sup.4 and R.sup.5, taken together with the carbon to which they
are both attached, form a carbocycle, substituted carbocycle,
heterocycle or substituted heterocycle; or R.sup.4 and R.sup.5,
when on the same carbon atom, taken together with the carbon to
which they are attached are --C(O)-- or --C(NR.sup.8)--; or two
R.sup.4 or two R.sup.5 on adjacent carbon atoms when taken together
with the carbons to which they are attached form a 3 to 6 membered
carbocycle, substituted carbocycle, heterocycle or substituted
heterocycle; R.sup.6 and R7 are each independently H, alkyl,
substituted alkyl, alkenyl, substituted alkenyl, alkynyl,
substituted alkynyl, haloalkyl, heteroalkyl, substituted
heteroalkyl, carbocyclyl, substituted carbocyclyl,
carbocyclylalkyl, substituted carbocyclylalkyl, heterocyclyl,
substituted heterocyclyl, heterocyclylalkyl, substituted
heterocyclylalkyl, arylalkyl, substituted arylalkyl,
heteroarylalkyl, substituted heteroarylalkyl,
carbocyclylheteroalkyl, substituted carbocyclylheteroalkyl,
heterocyclylheteroalkyl, substituted heterocyclylheteroalkyl,
arylheteroalkyl, substituted arylheteroalkyl,
heteroarylheteroalkyl, or substituted heteroarylheteroalkyl,
--C(O)H, --C(O)R.sup.8, --S(O)R.sup.8, --S(O).sub.2R.sup.8,
--C(O)OR.sup.8, or --C(O)NR.sup.9R.sup.10,
S(O).sub.2NR.sup.9R.sup.10; or R.sup.6 and R.sup.7, taken together
with the nitrogen to which they are both attached, form a
substituted or unsubstituted heterocycle, which may contain one or
more additional heteroatoms selected from N, O, P, or S; or R.sup.7
taken together with L.sup.2, and the N to which they are both
attached, forms a substituted or unsubstituted 3 to 8 membered
heterocycle which may contain one or more additional heteroatoms
selected from N, O, S, or P; R.sup.8 is H, alkyl, substituted
alkyl, haloalkyl, alkenyl, substituted alkenyl, alkynyl,
substituted alkynyl, heteroalkyl, substituted heteroalkyl,
carbocyclyl, substituted carbocyclyl, carbocyclylalkyl, substituted
carbocyclylalkyl, heterocyclyl, substituted heterocyclyl,
heterocyclylalkyl, substituted heterocyclylalkyl, arylalkyl,
substituted arylalkyl, heteroarylalkyl, substituted
heteroarylalkyl, carbocyclylheteroalkyl, substituted
carbocyclylheteroalkyl, heterocyclylheteroalkyl, substituted
heterocyclylheteroalkyl, arylheteroalkyl, substituted
arylheteroalkyl, heteroarylheteroalkyl, or substituted
heteroarylheteroalkyl; and R.sup.9 and R.sup.10 are each
independently H, alkyl, substituted alkyl, alkenyl, substituted
alkenyl, alkynyl, substituted alkynyl, haloalkyl, heteroalkyl,
substituted heteroalkyl, carbocyclyl, substituted carbocyclyl,
carbocyclylalkyl, substituted carbocyclylalkyl, heterocyclyl,
substituted heterocyclyl, heterocyclylalkyl, substituted
heterocyclylalkyl, arylalkyl, substituted arylalkyl,
heteroarylalkyl, substituted heteroarylalkyl,
carbocyclylheteroalkyl, substituted carbocyclylheteroalkyl,
heterocyclylheteroalkyl, substituted heterocyclylheteroalkyl,
arylheteroalkyl, substituted arylheteroalkyl,
heteroarylheteroalkyl, or substituted heteroarylheteroalkyl; or
R.sup.9 and R.sup.10, taken together with the nitrogen to which
they are both bonded, form a substituted or unsubstituted
heterocycle; wherein each substituted alkyl, substituted alkenyl,
substituted alkynyl, substituted heteroalkyl, substituted
carbocyclyl, substituted carbocyclylalkyl, substituted
heterocyclyl, substituted heterocyclylalkyl, substituted arylalkyl,
substituted heteroarylalkyl, substituted carbocyclylheteroalkyl,
substituted heterocyclylheteroalkyl, substituted arylheteroalkyl,
substituted heteroarylheteroalkyl, substituted alkylene,
substituted heteroalkylene, substituted alkenylene, substituted
alkynylene, substituted carbocyclylene, or substituted
heterocyclylene is independently substituted with one to four
substituents selected from the group consisting of -halogen, --R,
--O.sup.-, .dbd.O, --OR, --SR, --S.sup.-, --NR.sub.2,
--N(+)R.sub.3, .dbd.NR, --C(halogen).sub.3, --CR(halogen).sub.2,
--CR.sub.2(halogen), --CN, --OCN, --SCN, --N.dbd.C.dbd.O, --NCS,
--NO, --NO.sub.2, .dbd.N.sub.2, --N.sub.3, --NRC(.dbd.O)R,
--NRC(.dbd.O)OR, --NRC(.dbd.O)NRR, --C(.dbd.O)NRR, --C(.dbd.O)OR,
--OC(.dbd.O)NRR, OC(.dbd.O)OR, --C(.dbd.O)R, --S(.dbd.O).sub.2OR,
--S(.dbd.O).sub.2R, --OS(.dbd.O).sub.2OR, --S(.dbd.O).sub.2NR,
--S(.dbd.O)R, --NRS(.dbd.O).sub.2R, --NRS(.dbd.O).sub.2NRR,
--NRS(.dbd.O).sub.2OR, --OP(.dbd.O)(OR).sub.2,
--P(.dbd.O)(OR).sub.2, --P(O)(OR)(O)R, --C(.dbd.O)R, --C(.dbd.S)R,
--C(.dbd.O)OR, --C(.dbd.S)OR, --C(.dbd.C)SR, --C(.dbd.S)SR,
--C(.dbd.O)NRR, --C(.dbd.S)NRR, --C(.dbd.NR)NRR, and
--NRC(.dbd.NR)NRR; wherein each R is independently H, alkyl,
cycloalkyl, aryl, arylalkyl, or heterocyclyl. See US 20100143301
A1, the disclosure of which is incorporated by reference in its
entirety.
[0335] In some embodiments, the activating moiety is a TLR7 and/or
TLR8 agonist having the structure of:
##STR00074##
wherein:
L.sup.1 is --NH-- or --O--;
[0336] R.sup.1 is alkyl, substituted alkyl, heteroalkyl,
substituted heteroalkyl, heterocyclylalkyl, substituted
heterocyclylalkyl, carbocyclylalkyl or substituted
carbocyclylalkyl; each of R.sup.4 and R.sup.5 independently is H or
C.sub.1-C.sub.6 alkyl or R.sup.4 and R.sup.5 taken together with
the carbon to which they are attached is --C(O)--; X.sup.1 is
C.sub.1-C.sub.6 alkylene. C.sub.1-C.sub.6 heteroalkylene or
C.sub.1-C.sub.6 substituted heteroalkylene; D is phenyl, biphenyl
or pyridinyl, wherein said phenyl, biphenyl or pyridinyl is
substituted with -L.sup.2-NR.sup.6R.sup.7; or D is pyridinyl,
piperidinyl, piperazinyl or 1,2,3,4-tetrahydroisoquinolinyl; n is 0
or 1; R.sup.3 is halogen, cyano, alkyl, carbocyclyl,
carbocyclylalkyl, haloalkyl, --C(O)OR.sup.6, --C(O)NR.sup.9R.sup.10
or --CHO; L.sup.2 is C.sub.1-C.sub.6 alkylene or a covalent bond;
each of R.sup.6 and R.sup.7 independently is H, alkyl, or
heteroaryl; or R.sup.6 and R.sup.7 taken together with the nitrogen
to which they are attached form a substituted or unsubstituted 4-6
membered heterocycle comprising 0 to 2 heteroatoms selected from N,
O or S.
[0337] In some embodiments, the activating moiety is a TLR7 and/or
TLR8 agonist having the structure of:
##STR00075##
C. Amount of Immunotherapeutics in the Therapeutic Combinations
[0338] In another aspect, the present invention provides a
therapeutic combination comprising a target therapeutic and an
immunotherapeutic in an amount that is suitable for the combination
therapy treatment of diseases such as tumors and cancers.
[0339] In some embodiments, the immunotherapeutic is of an amount
that is capable of: (1) inducing IFN-.alpha. in a enriched human
blood DCs; (2) inducing TNF-.alpha. in a enriched human blood DCs;
and/or (3) inducing IL-12-.alpha. in a enriched human blood
DCs.
[0340] Methods for measuring the activity of the immunotherapeutics
are: 1) an assay to measure cytokines release from human dendritic
cell stimulated by immunotherapy; 2) an assay to detect antibody
dependent cell mediated cytotoxicity enhanced by immunotherapy; and
3) an efficacy study in a tumor model treated by immunotherapy.
[0341] In some embodiments, the immunotherapeutic (e.g. resiquimod
or its analogues) is adminstered, either orally or intravenously
using oral formulation or intravenous formulation, of an amount so
that the local concentration of the immunotherapeutics (e.g. near
or at the tumor site of a solid tumor) is between about 0.005
.mu.g/ml to about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 .mu.g/ml
(all inclusive).
[0342] The local concentration of the immunotherapeutics (e.g. near
or at the tumor site of a solid tumor) can measured using methods
known in the art, such as measuring the tissue or serum
concentration.
[0343] Local effective concentration of therapeutic agent is
depended on its absorption from various routes, tissue
distribution, and metabolism process, and plasma pharmacokinetics
of agent and tissue concentration could be measured routinely using
methods known in the art.
[0344] In some embodiments, the immunotherapeutic is adminstered of
an amount so that the local concentration of the immunotherapeutics
(e.g. near or at the tumor site of a solid tumor) is between about
0.05 .mu.g/ml, 0.1 .mu.g/ml, 0.15 .mu.g/ml, 0.2 .mu.g/ml, 0.3
.mu.g/ml, or 0.4 .mu.g/ml, to about 0.5 .mu.g/ml (all
inclusive).
[0345] In some embodiments, the subject is administered an oral
formulation comprising the immunotherapeutic (e.g. resiquimod or
its analogues) in a dose of between about 0.0005 mg/kg, 0.0006
mg/kg, 0.0007 mg/kg, 0.0008 mg/kg, 0.0009 mg/kg, 0.001 mg/kg, 0.002
mg/kg, 0.003 mg/kg, 0.004 mg/kg, 0.005 mg/kg, 0.006 mg/kg, 0.007
mg/kg, 0.008 mg/kg, 0.009 mg/kg, 0.01 mg/kg, or 0.015 mg/kg, to
about 0.02 mg/kg (all inclusive), two times per week. In some
embodiments, the subject is administered an oral formulation
comprising the immunotherapeutic (e.g. resiquimod or its analogues)
in a dose of between about 0.0005 mg/kg, to about 0.0006 mg/kg,
0.0007 mg/kg, 0.0008 mg/kg, 0.0009 mg/kg, 0.001 mg/kg, 0.002 mg/kg,
0.003 mg/kg, 0.004 mg/kg, 0.005 mg/kg, 0.006 mg/kg, 0.007 mg/kg,
0.008 mg/kg, 0.009 mg/kg, 0.01 mg/kg, 0.015 mg/kg, or 0.02 mg/kg
(all inclusive), two times per week.
[0346] In some embodiments, the subject is administered an oral
formulation comprising the immunotherapeutic (e.g. resiquimod or
its analogues) in a dose of less than or about 0.0005 mg/kg, 0.0006
mg/kg, 0.0007 mg/kg, 0.0008 mg/kg, 0.0009 mg/kg, 0.001 mg/kg, 0.002
mg/kg, 0.003 mg/kg, 0.004 mg/kg, 0.005 mg/kg, 0.006 mg/kg, 0.007
mg/kg, 0.008 mg/kg, 0.009 mg/kg, 0.01 mg/kg, two times per
week.
[0347] In some embodiments, the subject is administered an
intravenous formulation comprising the immunotherapeutic (e.g.
resiquimod or its analogues) in a dose of between about 0.0005
mg/kg, 0.0006 mg/kg, 0.0007 mg/kg, 0.0008 mg/kg, 0.0009 mg/kg,
0.001 mg/kg, 0.002 mg/kg, 0.003 mg/kg, 0.004 mg/kg, 0.005 mg/kg,
0.006 mg/kg, 0.007 mg/kg, 0.008 mg/kg, 0.009 mg/kg, 0.01 mg/kg, or
about 0.015 mg/kg, to about 0.02 mg/kg (inclusive), weekly. In some
embodiments, the subject is administered an intravenous formulation
comprising the immunotherapeutic (e.g. resiquimod or its analogues)
in a dose of between about 0.0005 mg/kg, to about 0.0006 mg/kg,
0.0007 mg/kg, 0.0008 mg/kg, 0.0009 mg/kg, 0.001 mg/kg, 0.002 mg/kg,
0.003 mg/kg, 0.004 mg/kg, 0.005 mg/kg, 0.006 mg/kg, 0.007 mg/kg,
0.008 mg/kg, 0.009 mg/kg, 0.01 mg/kg, 0.015 mg/kg, or 0.02 mg/kg
(inclusive), weekly.
[0348] In some embodiments, the subject is administered a
formulation comprising the immunotherapeutic (e.g. resiquimod or
its analogues) in a dose of between about 0.0005 mg/kg, 0.0006
mg/kg, 0.0007 mg/kg, 0.0008 mg/kg, 0.0009 mg/kg, 0.001 mg/kg, 0.002
mg/kg, 0.003 mg/kg, 0.004 mg/kg, 0.005 mg/kg, 0.006 mg/kg, 0.007
mg/kg, 0.008 mg/kg, 0.009 mg/kg, 0.01 mg/kg, 0.02 mg/kg, 0.025
mg/kg, 0.05 mg/kg, 0.075 mg/kg, 0.1 mg/kg, 0.125 mg/kg, 0.15 mg/kg,
0.175 mg/kg, 0.2 mg/kg, 0.215 mg/kg, 0.25 mg/kg, 0.5 mg/kg, 0.75
mg/kg, 1 mg/kg, 1.25 mg/kg, 1.5 mg/kg, 1.75 mg/kg, 2 mg/kg, 2.25
mg/kg, to about 2.5 mg/kg, all inclusive, twice daily, once daily,
once every two, three, four, five or six days, or once, twice, or
three times per week.
[0349] In some embodiments, the subject is administered a
formulation comprising the immunotherapeutic (e.g. resiquimod or
its analogues) in a dose of between about 0.0005 mg/kg, to about
0.0006 mg/kg, 0.0007 mg/kg, 0.0008 mg/kg, 0.0009 mg/kg, 0.001
mg/kg, 0.002 mg/kg, 0.003 mg/kg, 0.004 mg/kg, 0.005 mg/kg, 0.006
mg/kg, 0.007 mg/kg, 0.008 mg/kg, 0.009 mg/kg, 0.01 mg/kg, 0.02
mg/kg, 0.025 mg/kg, 0.05 mg/kg, 0.075 mg/kg, 0.1 mg/kg, 0.125
mg/kg, 0.15 mg/kg, 0.175 mg/kg, 0.2 mg/kg, 0.215 mg/kg, 0.25 mg/kg,
0.5 mg/kg, 0.75 mg/kg, 1 mg/kg, 1.25 mg/kg, 1.5 mg/kg, 1.75 mg/kg,
2 mg/kg, 2.25 mg/kg, or 2.5 mg/kg o, all inclusive, twice daily,
once daily, once every two, three, four, five or six days, or once,
twice, or three times per week.
[0350] In some embodiments, the subject is administered a
formulation comprising die immunotherapeutic (e.g. resiquimod or
its analogues) in a dose of less than or about 0.0005 mg/kg, 0.0006
mg/kg, 0.0007 mg/kg, 0.0008 mg/kg, 0.0009 mg/kg, 0.001 mg/kg, 0.002
mg/kg, 0.003 mg/kg, 0.004 mg/kg, 0.005 mg/kg, 0.006 mg/kg, 0.007
mg/kg, 0.008 mg/kg, 0.009 mg/kg, 0.01 mg/kg, 0.02 mg/kg, 0.025
mg/kg, 0.05 mg/kg, 0.075 mg/kg, 0.1 mg/kg, 0.125 mg/kg, 0.15 mg/kg,
0.175 mg/kg, 0.2 mg/kg, 0.215 mg/kg, 0.25 mg/kg, 0.5 mg/kg, 0.75
mg/kg, 1 mg/kg, 1.25 mg/kg, 1.5 mg/kg, 1.75 mg/kg, 2 mg/kg, 2.25
mg/kg, or 2.5 mg/kg, twice daily, once daily, once every two,
three, four, five or six days, or once, twice, or three times per
week.
[0351] In some embodiments, the administration is orally,
sublingually, intravenously, intramuscularly, subcutaneously, or
intratumorally. In some embodiments, the subject is administered a
formulation comprising the immunotherapeutic (e.g. resiquimod or
its analogues) in a dose of a range between any two doses selected
from the following doses: about 0.0005 mg/kg, 0.0006 mg/kg, 0.0007
mg/kg, 0.0008 mg/kg, 0.0009 mg/kg, 0.001 mg/kg, 0.002 mg/kg, 0.003
mg/kg, 0.004 mg/kg, 0.005 mg/kg, 0.006 mg/kg, 0.007 mg/kg, 0.008
mg/kg, 0.009 mg/kg, 0.01 mg/kg, 0.02 mg/kg, 0.025 mg/kg, 0.05
mg/kg, 0.075 mg/kg, 0.1 mg/kg, 0.125 mg/kg, 0.15 mg/kg, 0.175
mg/kg, 0.2 mg/kg, 0.215 mg/kg, 0.25 mg/kg, 0.5 mg/kg, 0.75 mg/kg, 1
mg/kg, 1.25 mg/kg, 1.5 mg/kg, 1.75 mg/kg, 2 mg/kg, 2.25 mg/kg, and
2.5 mg/kg.
[0352] In some embodiments, the method comprises administering to
said subject an intravenous formulation comprising said
immunotherapeutic (e.g. resiquimod or its analogues) in a dose of
between about from 0.0008 mg/kg to about 0.0133 mg/kg, weekly.
[0353] In some embodiments, the subject is administered an
intravenous formulation comprising the immunotherapeutic (e.g.
resiquimod or its analogues) in a dose of less than or about 0.003
mg/kg, 0.004 mg/kg, 0.005 mg/kg, or 0.006 mg/kg to about 0.007
mg/kg, weekly. For references regarding safe dosage of
immunotherapeutics, see Jurk et al., Nature Immunology, Vol. 4, No.
6''499 (2002), and Pockros et al., J. Hepatology, 47:174-182
(2007), the disclosure of which is incorporated by reference in
their entirety.
III. Pharmaceutical Formulations and Administration
[0354] The present invention further relates to a pharmaceutical
formulation comprising a compound of the invention or a
pharmaceutically acceptable salt thereof, and one or more
pharmaceutically acceptable carriers.
[0355] The compounds described herein including pharmaceutically
acceptable carriers such as addition salts or hydrates thereof can
be delivered to a patient using a wide variety of routes or modes
of administration. Suitable routes of administration include, but
inhalation, transdermal, oral, rectal, transmucosal, intestinal and
parenteral administration, including intramuscular, subcutaneous
and intravenous injections. Preferably, the compounds of the
invention comprising an antibody or antibody fragment as the
targeting moiety are administered parenterally, more preferably
intravenously.
[0356] As used herein, the terms "administering" or
"administration" are intended to encompass all means for directly
and indirectly delivering a compound to its intended site of
action.
[0357] The compounds described herein, or pharmaceutically
acceptable salts and/or hydrates thereof, may be administered
singly, in combination with other compounds of the invention,
and/or in cocktails combined with other therapeutic agents. Of
course, the choice of therapeutic agents that can be
co-administered with the compounds of the invention will depend, in
part, on the condition being treated.
[0358] For example, when administered to patients suffering from a
disease state caused by an organism that relies on an autoinducer,
the compounds of the invention can be administered in cocktails
containing agents used to treat the pain, infection and other
symptoms and side effects commonly associated with the disease.
Such agents include, e.g., analgesics, antibiotics, etc.
[0359] When administered to a patient undergoing cancer treatment,
the compounds may be administered in cocktails containing
anti-cancer agents and/or supplementary potentiating agents. The
compounds may also be administered in cocktails containing agents
that treat the side-effects of radiation therapy, such as
anti-emetics, radiation protectants, etc.
[0360] Supplementary potentiating agents that can be
co-administered with the compounds of the invention include, e.g.,
tricyclic anti-depressant drugs (e.g., imipramine, desipramine,
amitriptyline, clomipramine, trimipramine, doxepin, nortriptyline,
protriptyline, amoxapine and maprotiline); non-tricyclic and
anti-depressant drugs (e.g., sertraline, trazodone and citalopram);
Ca+2 antagonists (e.g., verapamil, nifedipine, nitrendipine and
caroverine); amphotericin; triparanol analogues (e.g., tamoxifen);
antiarrhythmic drugs (e.g., quinidine); antihypertensive drugs
(e.g., reserpine); thiol depleters (e.g., buthionine and
sulfoximine); and calcium leucovorin.
[0361] The active compound(s) of the invention are administered per
se or in the form of a pharmaceutical composition wherein the
active compound(s) is in admixture with one or more
pharmaceutically acceptable carriers, excipients or diluents.
Pharmaceutical compositions for use in accordance with the present
invention are typically formulated in a conventional manner using
one or more physiologically acceptable carriers comprising
excipients and auxiliaries, which facilitate processing of the
active compounds into preparations which, can be used
pharmaceutically. Proper formulation is dependent upon the route of
administration chosen.
[0362] For transmucosal administration, penetrants appropriate to
the barrier to be permeated are used in the formulation. Such
penetrants are generally known in the art.
[0363] For oral administration, the compounds can be formulated
readily by combining the active compound(s) with pharmaceutically
acceptable carriers well known in the art. Such carriers enable the
compounds of the invention to be formulated as tablets, pills,
dragees, capsules, liquids, gels, syrups, slurries, and suspensions
for oral ingestion by a patient to be treated. Pharmaceutical
preparations for oral use can be obtained solid excipient,
optionally grinding a resulting mixture, and processing the mixture
of granules, after adding suitable auxiliaries, if desired to
obtain tablets or dragee cores. Suitable excipients are, in
particular, fillers such as sugars, including lactose, sucrose,
mannitol, or sorbitol; cellulose preparations such as, for example,
maize starch, wheat starch, rice starch, potato starch, gelatin,
gum tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose,
sodium carboxymethylcellulose, and/or polyvinylpyrrolidone (PVP).
If desired, disintegrating agents may be added, such as the
cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt
thereof such as sodium alginate.
[0364] Dragee cores are provided with suitable coatings. For this
purpose, concentrated sugar solutions may be used, which may
optionally contain gum arabic, talc, polyvinyl pyrrolidone,
carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquer
solutions, and suitable organic solvents or solvent mixtures.
Dyestuffs or pigments may be added to the tablets or dragee
coatings for identification or to characterize different
combinations of active compound doses.
[0365] Pharmaceutical preparations, which can be used orally,
include push-fit capsules made of gelatin, as well as soft, sealed
capsules made of gelatin and a plasticizer, such as glycerol or
sorbitol. The push-fit capsules can contain the active ingredients
in admixture with filler such as lactose, binders such as starches,
and/or lubricants such as talc or magnesium stearate and,
optionally, stabilizers. In soft capsules, the active compounds may
be dissolved or suspended in suitable liquids, such as fatty oils,
liquid paraffin, or liquid polyethylene glycols. In addition,
stabilizers may be added. All formulations for oral administration
should be in dosages suitable for such administration.
[0366] For buccal administration, the compositions may take the
form of tablets or lozenges formulated in conventional manner.
[0367] For administration by inhalation, the compounds for use
according to the present invention are conveniently delivered in
the form of an aerosol spray presentation from pressurized packs or
a nebulizer, with the use of a suitable propellant e.g.,
dichlorodifluoromethane, trichlorofluoromethane,
dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In
the case of a pressurized aerosol the dosage unit may be determined
by providing a valve to deliver a metered amount. Capsules and
cartridges of e.g., gelatin for use in an inhaler or insufflator
may be formulated containing a powder mix of the compound and a
suitable powder base such as lactose or starch.
[0368] The compounds may be formulated for parenteral
administration by injection, e.g., by bolus injection or continuous
infusion. Injection is a preferred method of administration for the
compositions of the current invention. Formulations for injection
may be presented in unit dosage form, e.g., in ampoules or in
multi-dose containers, with an added preservative. The compositions
may take such forms as suspensions, solutions or emulsions in oily
or aqueous vehicles, and may contain formulatory agents such as
suspending, stabilizing and/or dispersing agents may be added, such
as the cross-linked polyvinyl pyrrolidone, agar, or alginic acid or
a salt thereof such as sodium alginate.
[0369] Pharmaceutical formulations for parenteral administration
include aqueous solutions of the active compounds in water-soluble
form. Additionally, suspensions of the active compounds may be
prepared as appropriate oily injection suspensions. Suitable
lipophilic solvents or vehicles include fatty oils such as sesame
oil, or synthetic fatty acid esters, such as ethyl oleate or
triglycerides, or liposomes. Aqueous injection suspensions may
contain substances, which increase the viscosity of the suspension,
such as sodium carboxymethyl cellulose, sorbitol, or dextran.
Optionally, the suspension may also contain suitable stabilizers or
agents, which increase the solubility of the compounds to allow for
the preparation of highly, concentrated solutions. For injection,
the agents of the invention may be formulated in aqueous solutions,
preferably in physiologically compatible buffers such as Hanks's
solution, Ringer's solution, or physiological saline buffer.
[0370] Alternatively, the active ingredient may be in powder form
for constitution with a suitable vehicle, e.g., sterile
pyrogen-free water, before use.
[0371] The compounds may also be formulated in rectal compositions
such as suppositories or retention enemas, e.g., containing
conventional suppository bases such as cocoa butter or other
glycerides.
[0372] In addition to the formulations described previously, the
compounds may also be formulated as a depot preparation. Such long
acting formulations may be administered by implantation or
transcutaneous delivery (e.g., subcutaneously or intramuscularly),
intramuscular injection or a transdermal patch. Thus, for example,
the compounds may be formulated with suitable polymeric or
hydrophobic materials (e.g., as an emulsion in an acceptable oil)
or ion exchange resins, or as sparingly soluble derivatives, for
example, as a sparingly soluble salt.
[0373] The pharmaceutical compositions also may comprise suitable
solid or gel phase carriers or excipients. Examples of such
carriers or excipients include calcium carbonate, calcium phosate,
various sugars, starches, cellulose derivatives, gelatin, and
polymers such as polyethylene glycols.
[0374] A preferred pharmaceutical composition is a composition
formulated for injection such as intravenous injection and includes
about 0.01% to about 100% by weight of the compound of the present
invention, based upon 100% weight of total pharmaceutical
composition. The drug-ligand conjugate may be an antibody-cytotoxin
conjugate where the antibody has been selected to target a
particular cancer.
[0375] In some embodiments, the pharmaceutical composition of the
present invention further comprises an additional therapeutic
agent.
[0376] In some embodiments, the additional therapeutic agent is an
anticancer agent.
[0377] In some embodiments, the additional anticancer agent is
selected from an antimetabolite, an inhibitor of topoisomerase I
and II, an alkylating agent, a microtubule inhibitor, an
antiandrogen agent, a GNRh modulator or mixtures thereof.
[0378] In some embodiments, the additional therapeutic agent is a
chemotherapeutic agent.
[0379] By "chemotherapeutic agent" herein is meant a chemical
compound useful in the treatment of cancer. Examples are but not
limited to: Gemcitabine, Irinotecan, Doxorubicin, 5-Fluorouracil,
Cytosine arabinoside ("Ara-C"), Cyclophosphamide, Thiotepa,
Busulfan, Cytoxin, TAXOL, Methotrexate, Cisplatin, Melphalan,
Vinblastine and Carboplatin.
[0380] In some embodiments, the second chemotherapeutic agent is
selected from the group consisting of tamoxifen, raloxifene,
anastrozole, exemestane, letrozole, imatanib, paclitaxel,
cyclophosphamide, lovastatin, minosine, gemcitabine, cytarabine,
5-fluorouracil, methotrexate, docetaxel, goserelin, vincristine,
vinblastine, nocodazole, teniposide etoposide, gemcitabine,
epothilone, vinorelbine, camptothecin, daunorubicin, actinomycin D,
mitoxantrone, acridine, doxorubicin, epirubicin, or idarubicin.
IV. Kits
[0381] In another aspect, the present invention provides kits
containing the therapeutic combinations provided herein and
directions for using the therapeutic combinations. The kit may also
include a container and optionally one or more vial, test tube,
flask, bottle, or syringe. Other formats for kits will be apparent
to those of skill in the art and are within the scope of the
present invention.
V. Medical Use
[0382] In another aspect, the present invention provides a method
for treating a disease condition in a subject that is in need of
such treatment, comprising: administering to the subject a
therapeutic combination or pharmaceutical composition comprising a
therapeutically effective amount of the compound of the present
invention or a pharmaceutically acceptable salt thereof, and a
pharmaceutical acceptable carrier.
[0383] In addition to the compositions and constructs described
above, the present invention also provides a number of uses of the
combinations of the invention. Uses of the combinations of the
current invention include: killing or inhibiting the growth,
proliferation or replication of a tumor cell or cancer cell,
treating cancer, treating a pre-cancerous condition, preventing the
multiplication of a tumor cell or cancer cell, preventing cancer,
preventing the multiplication of a cell that expresses an
autoimmune antibody. These uses comprise administering to an animal
such as a mammal or a human in need thereof an effective amount of
a compound of the present invention.
[0384] The combination of the current invention is useful for
treating diseases such as cancer in a subject, such as a human
being. Combinations and uses for treating tumors by providing a
subject the composition in a pharmaceutically acceptable manner,
with a pharmaceutically effective amount of a composition of the
present invention are provided.
[0385] By "cancer" herein is meant the pathological condition in
humans that is characterized by unregulated cell proliferation.
Examples include but are not limited to: carcinoma, lymphoma,
blastoma, and leukemia. More particular examples of cancers include
but are not limited to: lung (small cell and non-small cell),
breast, prostate, carcinoid, bladder, gastric, pancreatic, liver
(hepatocellular), hepatoblastoma, colorectal, head and neck
squamous cell carcinoma, esophageal, ovarian, cervical,
endometrial, mesothelioma, melanoma, sarcoma, osteosarcoma,
liposarcoma, thyroid, desmoids, chronic myelocytic leukemia (AML),
and chronic myelocytic leukemia (CML).
[0386] By "inhibiting" or "treating" or "treatment" herein is meant
to reduction, therapeutic treatment and prophylactic or
preventative treatment, wherein the objective is to reduce or
prevent the aimed pathologic disorder or condition. In one example,
following administering of a compound of the present invention, a
cancer patient may experience a reduction in tumor size.
"Treatment" or "treating" includes (1) inhibiting a disease in a
subject experiencing or displaying the pathology or symptoms of the
disease, (2) ameliorating a disease in a subject that is
experiencing or displaying the pathology or symptoms of the
disease, and/or (3) affecting any measurable decrease in a disease
in a subject or patient that is experiencing or displaying the
pathology or symptoms of the disease. To the extent a compound of
the present invention may prevent growth and/or kill cancer cells,
it may be cytostatic and/or cytotoxic.
[0387] By "therapeutically effective amount" herein is meant an
amount of a compound provided herein effective to "treat" a
disorder in a subject or mammal. In the case of cancer, the
therapeutically effective amount of the drug may either reduce the
number of cancer cells, reduce the tumor size, inhibit cancer cell
infiltration into peripheral organs, inhibit tumor metastasis,
inhibit tumor growth to certain extent, and/or relieve one or more
of the symptoms associated with the cancer to some extent.
[0388] Administration "in combination with" one or more further
therapeutic agents includes simultaneous (concurrent) and
consecutive administration in any order. As used herein, the term
"pharmaceutical combination" refers to a product obtained from
mixing or combining active ingredients, and includes both fixed and
non-fixed combinations of the active ingredients. The term "fixed
combination" means that the active ingredients, e.g. a compound of
Formula (1) and a co-agent, are both administered to a patient
simultaneously in the form of a single entity or dosage. The term
"non-fixed combination" means that the active ingredients, e.g. a
compound of Formula (1) and a co-agent, are both administered to a
patient as separate entities either simultaneously, concurrently or
sequentially with no specific time limits, wherein such
administration provides therapeutically effective levels of the
active ingredients in the body of the patient. The latter also
applies to cocktail therapy, e.g. the administration of three or
more active ingredients.
[0389] In some embodiments, the diseases condition is tumor or
cancer. In some embodiments, the cancer or tumor is selected from
stomach, colon, rectal, liver, pancreatic, lung, breast, cervix
uteri, corpus uteri, ovary, testis, bladder, renal, brain/CNS, head
and neck, throat, Hodgkin's disease, non-Hodgkin's lymphoma,
multiple myeloma, leukemia, melanoma, non-melanoma skin cancer,
acute lymphocytic leukemia, acute myelogenous leukemia, Ewing's
sarcoma, small cell lung cancer, choriocarcinoma, rhabdomyosarcoma,
Wilms' tumor, neuroblastoma, hairy cell leukemia, mouth/pharynx,
oesophagus, larynx, kidney cancer or lymphoma.
[0390] In some embodiments, the disease condition comprises
abnormal cell proliferation, such as a pre-cancerous lesion.
[0391] The current invention is particularly useful for the
treatment of cancer and for the inhibition of the multiplication of
a tumor cell or cancer cell in an animal. Cancer, or a precancerous
condition, includes a tumor, metastasis, or any disease or disorder
characterized by uncontrolled cell growth, can be treated or
prevented by administration the drug-ligand complex of the current
invention. The compound delivers the activating moiety to a tumor
cell or cancer cell. In some embodiments, the targeting moiety
specifically binds to or associates with a cancer-cell or a
tumor-cell-associated antigen. Because of its close proximity to
the ligand, after being internalized, the activating moiety can be
taken up inside a tumor cell or cancer cell through, for example,
receptor-mediated endocytosis. The antigen can be attached to a
tumor cell or cancer cell or can be an extracellular matrix protein
associated with the tumor cell or cancer cell. Once inside the
cell, the linker is hydrolytically or enzymatically cleaved by a
tumor-cell or cancer-cell-associated proteases, thereby releasing
the activating moiety. The released activating moiety is then free
to diffuse and induce or enhance immune activity of immune cells or
tumor cells. In an alternative embodiment, the activating moiety is
cleaved from the compound tumor microenvironment, and the drug
subsequently penetrates the cell.
[0392] Representative examples of precancerous conditions that may
be targeted by the compounds of the present invention, include:
metaplasia, hyperplasia, dysplasia, colorectal polyps, actinic
ketatosis, actinic cheilitis, human papillomaviruses, leukoplakia,
lychen planus and Bowen's disease.
[0393] Representative examples of cancers or tumors that may be
targeted by compounds of the present invention include: lung
cancer, colon cancer, prostate cancer, lymphoma, melanoma, breast
cancer, ovarian cancer, testicular cancer, CNS cancer, renal
cancer, kidney cancer, pancreatic cancer, stomach cancer, oral
cancer, nasal cancer, cervical cancer and leukemia. It will be
readily apparent to the ordinarily skilled artisan that the
particular targeting moiety used in the compound can be chosen such
that it targets the activating moiety to the tumor tissue to be
treated with the drug (i.e., a targeting agent specific for a
tumor-specific antigen is chosen). Examples of such targeting
moiety are well known in die art, examples of which include
anti-Her2 for treatment of breast cancer, anti-CD20 for treatment
of lymphoma, anti-PSMA for treatment of prostate cancer and
anti-CD30 for treatment of lymphomas, including non-Hodgkin's
lymphoma.
[0394] In some embodiments, the abnormal proliferation is of cancer
cells.
[0395] In some embodiments, the cancer is selected from the group
consisting of: breast cancer, colorectal cancer, diffuse large
B-cell lymphoma, endometrial cancer, follicular lymphoma, gastric
cancer, glioblastoma, head and neck cancer, hepatocellular cancer,
lung cancer, melanoma, multiple myeloma, ovarian cancer, pancreatic
cancer, prostate cancer, and renal cell carcinoma.
[0396] In some embodiments, the present invention provides a
compound for use in killing a cell. The compound is administered to
the cell in an amount sufficient to kill said cell. In an
exemplary-embodiment, the compound is administered to a subject
bearing the cell. In a further exemplary embodiment, the
administration serves to retard or stop the growth of a tumor that
includes the cell (e.g., the cell can be a tumor cell). For the
administration to retard the growth, the rate of growth of the cell
should be at least 10% less than the rate of growth before
administration. Preferably, the rate of growth will be retarded at
least 20% 30%, 40%, 50%, 60%, 70%, 80%, 90%, or completely
stopped.
[0397] Additionally, the present invention provides a compound or a
pharmaceutical composition of the present invention for use as a
medicament. The present invention also provides a compound or a
pharmaceutical composition for killing, inhibiting or delaying
proliferation of a tumor or cancer cell, or for treating a disease
wherein TLR7 and/or TLR8 are implicated.
Effective Dosages
[0398] Pharmaceutical compositions suitable for use with the
present invention include compositions wherein the active
ingredient is contained in a therapeutically effective amount,
i.e., in an amount effective to achieve its intended purpose. The
actual amount effective for a particular application will depend,
inter alia, on the condition being treated. Determination of an
effective amount is well within the capabilities of those skilled
in the art, especially in light of the detailed disclosure
herein.
[0399] For any compound described herein, the therapeutically
effective amount can be initially determined from cell culture
assays. Target plasma concentrations will be those concentrations
of active compound(s) that are capable of inhibition cell growth or
division. In preferred embodiments, the cellular activity is at
least 25% inhibited. Target plasma concentrations of active
compound(s) that are capable of inducing at least about 30%, 50%,
75%, or even 90% or higher inhibition of cellular activity are
presently preferred. The percentage of inhibition of cellular
activity in the patient can be monitored to assess the
appropriateness of the plasma drug concentration achieved, and the
dosage can be adjusted upwards or downwards to achieve the desired
percentage of inhibition.
[0400] As is well known in the art, therapeutically effective
amounts for use in humans can also be determined from animal
models. For example, a dose for humans can be formulated to achieve
a circulating concentration that has been found to be effective in
animals. The dosage in humans can be adjusted by monitoring
cellular inhibition and adjusting the dosage upwards or downwards,
as described above.
[0401] A therapeutically effective dose can also be determined from
human data for compounds which are known to exhibit similar
pharmacological activities. The applied dose can be adjusted based
on the relative bioavailability and potency of the administered
compound as compared with the known compound.
[0402] Adjusting the dose to achieve maximal efficacy in humans
based on the methods described above and other methods as are
well-known in the art is well within the capabilities of the
ordinarily skilled artisan.
[0403] In some embodiments, the composition of the present
invention is delivered local or regional to a tumor located in the
subject, delivered systemically, or delivered via intratumoral
injection or by direct injection into tumor vasculature.
[0404] In the case of local administration, the systemic
circulating concentration of administered compound will not be of
particular importance. In such instances, the compound is
administered so as to achieve a concentration at the local area
effective to achieve the intended result.
[0405] Therapeutic amounts of specific antibodies disclosed herein
can also be administered, as a component of the combination, with
the immunotherapeutics, either in a single mixture form, or
separately. In some embodiments, therapeutic amounts are amounts
which eliminate or reduce the patient's tumor burden, or which
prevent or reduce the proliferation of metastatic cells. The dosage
will depend on many parameters, including the nature of the tumor,
patient history, patient condition, the possible co-use of other
oncolytic agents, and methods of administration. Methods of
administration include injection (e.g., parenteral, subcutaneous,
intravenous, intraperitoneal, etc.) for which the antibodies are
provided in a nontoxic pharmaceutically acceptable carrier such as
water, saline, Ringer's solution, dextrose solution, 5% human serum
albumin, fixed oils, ethyl oleate, or liposomes. Typical dosages
may range from about 0.01 to about 20 mg/kg, such as from about 0.1
to about 10 mg/kg. Other effective methods of administration and
dosages may be determined by routine experimentation and are within
the scope of this invention.
[0406] The therapeutically effective amount of the agents
(disclosed herein) administered, when it is used for combination
therapy, can vary depending upon the desired effects and the
subject to be treated. For example, the subject can receive at
least 1 mg/kg (such as 1 mg/kg to 20 mg/kg, 2.5 mg/kg to 10 mg/kg,
or 3.75 mg/kg to 5 mg/kg) intravenously of each antibody agent. The
dosage can be administered in divided doses (such as 2, 3, or 4
divided doses per day), or in a single dosage.
[0407] In the method for combined administration, the agent may be
simultaneously administered with the antibody used in the present
invention, or the agent may be administered before or after the
administration of the antibody used in the present invention.
[0408] For other modes of administration, dosage amount and
interval can be adjusted individually to provide plasma levels of
the administered compound effective for the particular clinical
indication being treated. For example, in one embodiment, a
compound according to the invention can be administered in
relatively high concentrations multiple times per day.
Alternatively, it may be more desirable to administer a compound of
the invention at minimal effective concentrations and to use a less
frequent administration regimen. This will provide a therapeutic
regimen that is commensurate with the severity of the individual's
disease.
[0409] Utilizing the teachings provided herein, an effective
therapeutic treatment regimen can be planned which does not cause
substantial toxicity and yet is entirely effective to treat the
clinical symptoms demonstrated by the particular patient. This
planning should involve the careful choice of active compound by
considering factors such as compound potency, relative
bioavailability, patient body weight, presence and severity of
adverse side effects, preferred mode of administration and the
toxicity-profile of the selected agent.
[0410] While preferred embodiments of the present invention have
been shown and described herein, it will be obvious to those
skilled in the art that such embodiments are provided by way of
example only. Numerous variations, changes, and substitutions will
now occur to those skilled in the art without departing from the
invention. It should be understood that various alternatives to the
embodiments of the invention described herein may be employed in
practicing the invention. It is intended that the following claims
define the scope of the invention and that methods and structures
within the scope of these claims and their equivalents be covered
thereby.
EXAMPLES
[0411] The present invention is further exemplified, but not
limited, by the following and Examples that illustrate the
preparation of the compounds of the invention.
Example 1
[0412] Tumour Inoculation and Evaluation of Tumour Growth
[0413] Mice: Female 6-week-old BALB/c and C3H/HeN (C3H) mice were
purchased from Japan SLC (Hamamatsu, Japan). All procedures were
reviewed and approved by the Animal Care and Use Committee of the
Tokyo Medical and Dental University. The SCCVII (C3H-originated,
3.times.10.sup.5), or Colon26 (BALB/c-originated, 5.times.10.sup.5)
parental cells were injected subcutaneously (s.c.) into the shaved
right flank of syngeneic mice and tumour volumes were evaluated. In
experiments examining the effects of anti-PDL1 (MIH5) mAb with
TLRL, 200 ug of anti-PDL1 mAb or 200 ug of anti-PDL1 mAb mixture
with TLRL or control rat IgG was injected i.p. three times a week
after tumour inoculation. Tumor volumes were measured along three
orthogonal axes (x, y, and z) and calculated as tumor
volume=(xyz)/2, if a mouse lose more than 20% of body weight or is
very sick and cannot get to adequate food or water, it will be
removed from the study and euthanized. (FIGS. 1 and 2)
Example 2
[0414] Enrichment of Human Dendritic Cells (DCs) from PBMC
[0415] Human PBMC was prepared from Buffy coats obtained from
healthy volunteer donors by Ficoll centrifugation. Dendritic cells
were enriched by using negative depletion with magnetic beads
(Miltenyi Biotec Inc. San Diego, Calif.) with mixture of anti-CD3,
CD19, CD20, CD14, and CD16 antibodies from human PBMC. The
enrichment of DCs was stained with goat anti-mouse FITC (lineages),
HLA-DR-APCCy7, CD123-BV421 and CD11C-APC. The stained cells were
analyzed on BD LSR Fortessa (BD Biosciences). The anti-CD3, CD4,
CD11C, CD19, CD14, CD16, CD123 monoclonal antibody were purchased
from BD Biosciences, CA or Biolegend, San Diego, Calif.
[0416] Stimulation of Enriched Human DCs and Cytokines
Expression
[0417] 1-2.times.10.sup.5 enriched DCs were plated in a 96-well
plate in 100 .mu.l media, 100 .mu.l diluted stimulators (including
TLRL were add to the plate and cultured for 20-22 h in 37.degree.
C. incubator. The supernatant were collected and human IFN-.alpha.,
IL-12(p70) and TNF-.alpha. were analyzed by ELISA (Mabtech AB,
Sweden).
[0418] FIGS. 3A-3G depict analysis of cytokine production by
enriched human DCs from three healthy donors. Enriched human DCs
were plated in a 96-well plate and cultured with allogeneic
untreated (medium) or treated different concentration of TLRL
directly for 20-22 h in 37.degree. C. incubator. The supernatant
were collected and human IFN-.alpha., IL-12(p70) and TNF-.alpha.
were analyzed by ELISA. Data are given as mean.+-.SD of triplicate
cultures. Three independent experiments from three healthy donors
were performed (Donor 1: FIG. 3A, Donor 2: FIG. 3B-D; Donor 3: FIG.
3E-G).
Example 3
[0419] Detection of Systemic Immune Activation with IFN Inducible
Genes Expression in Mouse PBMC by TLRL
[0420] Balb/c mice, 6-8 weeks of age, female, purchased from Vital
River were injected intravenously with TLRL, at indicated time
point, mice were bled and IFN inducible genes were examined by
qPCR. Once pick time of expression IFN inducible genes was
determined, a separated experiment was performance with various
dose of TLRL. At indicated time point, mice were bled and IFN
inducible genes were examined. The Quantitative Real-Time PCR was
performed and gene expression data were normalized relative to
geometric mean of two housekeeping genes (Actin):
TABLE-US-00002 Mouse Actin: F: (SEQ ID NO.: 1)
CATTGCTGACAGGATGCAGAAGG, Mouse Actin R: (SEQ ID NO.: 2)
TGCTGGAAGGTGGACAGTGAGG; Mouse Inf-b: F: (SEQ ID NO.: 3)
CTCCAGCACTGGGTGGAATG, Mouse Inf-b R: (SEQ ID NO.: 4)
AGTGGAGAGCAGTTGAGGAC; Mouse Mx2: F; (SEQ ID NO.: 5)
GTGGCAGAGGGAGAATGTCG, Mouse Mx2 R: (SEQ ID NO.: 6)
TAAAACAGCATAACCTTTTGCGA; Mouse Ifn-a: F: (SEQ ID NO.: 7)
CCTGAGAGAGAAGAAACACAGCC, Mouse Ifn-a R: (SEQ ID NO.: 8)
GGCTCTCCAGACTTCTGCTCTG; Mouse ISG15: F: (SEQ ID NO.: 9)
CAGCAATGGCCTGGGACCTAA, Mouse ISG15R: (SEQ ID NO.: 10)
GGAAAGCCGGCACACCAATC.
[0421] FIGS. 4A-4C depict expression of IFN inducible genes in
mouse PBMC after TLRL injection. RNA was isolated from PBMCs
cryopreserved with TRIzol reagent at variable time points and
Relative expression of IFN inducible genes were determined by
quantitative RT-PCR. MX2 gene was detected over time course of 5
hours post TLRL injection (FIG. 4A) and MX2 and ISG15 genes were
measured with various dose of TLRL at 2 hours post injection (FIG.
4B and FIG. 4C). Values indicate the mRNA expression of indicated
IFN inducible genes relative to housekeeping gene Actin. Bar graphs
represent data from 3 individual animals. **P<0.01;
***P<0.001.
[0422] Statistical Analysis
[0423] The significance of all comparisons was calculated using a
Student's two-tailed t test assuming unequal variance between mock
and sample groups, and results considered significant when
p<0.05. Correlations between parameters were assessed using
Spearman's rank correlation test, P values <0.05 were consider
to be statistically significant.
Example 4
[0424] PD-L1 Expressed C26 Tumor Model
[0425] Combinational Efficacy of R848 and Anti-PD1 Antibody in
Immune-Competent Mouse of PD-L1 Expressed C26 Tumor Model
[0426] The objective of this study was to determine the single or
combinational efficacy of R848 and anti-PD-1 monoclonal antibody in
the mouse colon adenocarcinoma 26 (C26) tumor model in
immune-competent mouse. C26 cells were inoculated by subcutaneous
injection into the right flank of BALB/c mice on Day 0. Tumors were
monitored until they reached a mean tumor volume of between 60 and
100 mm3. Mice were then administered over a three-week period 10
mg/kg IV (via tail vein) of rat anti-mouse PD-1 monoclonal antibody
alone twice weekly, 0.08 mg/kg of R848 alone once weekly, or a
combination of anti-PD-1 antibody and R848. Tumor volumes were
recorded twice weekly over the course of the study and the mice
were observed regularly. Tumor-bearing mice (treatment, control)
were sacrificed at Day 26 or if they became moribund before the
tumor volume reached 2000 mm3. Limited inhibition in tumor growth
was detected following treatment with R848 alone or anti-PD-1
antibody alone. The antitumor activity of R848 alone or anti-PD-1
antibody alone was markedly enhanced when given in combination.
Following combination treatment with R848 and anti-PD-1 antibody,
tumor size was greatly reduced with 83.75% tumor inhibition. The
differences in tumor inhibition by the combination treatment
compared to that of each component is shown graphically (FIG. 5)
and was highly significant (p=0.0002, anti-PD-1 and R848 combined
versus R848 alone; p=0.0063 anti-PD-1 and R848 combined versus
anti-PD-1 alone). No adverse effects on clinical signs, body weight
or survival were observed in animals following treatment with the
combination of R848 and anti-PD-1 antibody.
[0427] Combinational Efficacy of Anti-PD-L1 and R848 in
Immune-Competent Mouse of PD-L1 Expressed C26 Tumor Model (Study
No. R-BDB001-0008)
[0428] The objective of this study was to determine the single or
combinational efficacy of R848 and anti-PD-L1 monoclonal antibody
in the C26 tumor model in immune-competent mouse. C26 cells were
inoculated by subcutaneous injection into the right flank of BALB/c
mice on Day 0. Tumors were monitored until they reached a mean
tumor volume of between 60 and 100 mm3, inclusive. Mice were
administered, over a 3-week period, 10 mg/kg IV (via tail vein) of
rat anti-mouse PD-L1 monoclonal antibody alone twice weekly, 0.08
mg/kg of R848 alone once weekly, or a combination of anti-PD-L1
antibody and R848.
[0429] Limited inhibition in tumor growth was detected following
treatment with R848 alone or anti-PD-L1 antibody alone. The
antitumor activity of R848 alone or anti-PD-L1 antibody alone was
markedly enhanced when given in combination. Following combination
treatment with R848 and anti-PD-L1 antibody, tumor size was greatly
reduced with 96% tumor inhibition. The differences in tumor
inhibition by the combination treatment compared to that of each
component is shown graphically (FIG. 6)) and was highly significant
(p=0.0029, anti-PDL1 and R848 combined versus R848 alone; p=0.0074,
anti-PDL1 and R848 combined versus anti-PDL1 alone). No adverse
effects on clinical signs, body weight or survival were observed in
animals following treatment with the combination of R848 and
anti-PD-L1 antibody.
Sequence CWU 1
1
10123DNAArtificial SequenceMouse actin forward primer 1cattgctgac
aggatgcaga agg 23222DNAArtificial SequenceMouse actin reverse
primer 2tgctggaagg tggacagtga gg 22320DNAArtificial SequenceMouse
Inf-b forward primer 3ctccagcact gggtggaatg 20420DNAArtificial
SequenceMouse Inf-b reverse primer 4agtggagagc agttgaggac
20520DNAArtificial SequenceMouse Mx2 forward primer 5gtggcagagg
gagaatgtcg 20623DNAArtificial SequenceMouse Mx2 reverse primer
6taaaacagca taaccttttg cga 23723DNAArtificial SequenceMouse Ifn-a
forward primer 7cctgagagag aagaaacaca gcc 23822DNAArtificial
SequenceMouse Ifn-a reverse primer 8ggctctccag acttctgctc tg
22921DNAArtificial SequenceMouse ISG15 forward primer 9cagcaatggc
ctgggaccta a 211020DNAArtificial SequenceMouse ISG15 reverse primer
10ggaaagccgg cacaccaatc 20
* * * * *