U.S. patent application number 15/713813 was filed with the patent office on 2018-02-01 for combinations of albumin-based drug delivery systems.
The applicant listed for this patent is KTB TUMORFORSCHUNGSGESELLSCHAFT MBH. Invention is credited to Felix Kratz, Andre Warnecke.
Application Number | 20180028679 15/713813 |
Document ID | / |
Family ID | 47988875 |
Filed Date | 2018-02-01 |
United States Patent
Application |
20180028679 |
Kind Code |
A1 |
Kratz; Felix ; et
al. |
February 1, 2018 |
Combinations Of Albumin-Based Drug Delivery Systems
Abstract
The present invention relates to a composition comprising at
least two different albumin-based drug delivery systems, as well as
to a pharmaceutical composition comprising said composition.
Inventors: |
Kratz; Felix; (Ehrenkirchen,
DE) ; Warnecke; Andre; (Freiburg, DE) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
KTB TUMORFORSCHUNGSGESELLSCHAFT MBH |
Freiburg Im Breisgau |
|
DE |
|
|
Family ID: |
47988875 |
Appl. No.: |
15/713813 |
Filed: |
September 25, 2017 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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14380190 |
Aug 21, 2014 |
9801949 |
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PCT/EP2013/000513 |
Feb 21, 2013 |
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15713813 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 31/519 20130101;
A61K 31/704 20130101; A61P 35/00 20180101; A61K 2300/00 20130101;
A61K 2300/00 20130101; A61K 38/07 20130101; A61K 31/519 20130101;
A61K 47/64 20170801; A61K 31/704 20130101; A61K 47/643
20170801 |
International
Class: |
A61K 38/07 20060101
A61K038/07; A61K 31/704 20060101 A61K031/704; A61K 31/519 20060101
A61K031/519 |
Foreign Application Data
Date |
Code |
Application Number |
Feb 21, 2012 |
EP |
12001136.6 |
Claims
1. A method of treating a disease comprising: administering an
effective amount of a pharmaceutical composition comprising a
combination of at least two different albumin-based drug delivery
systems, wherein at least one of said at least two different
albumin-based drug delivery systems is selected from
albumin-binding prodrugs, albumin drug conjugates, albumin peptide
conjugates, albumin fusion proteins, albumin-binding peptide
conjugates, albumin drug nanoparticles, and albumin-based antibody
constructs and a pharmaceutically acceptable carrier and/or a
pharmaceutically acceptable adjuvant and/or diluent, wherein the
disease is selected from the group consisting of cancer, autoimmune
disease, acute or chronic inflammatory diseases, or diseases caused
by viruses and/or microorganisms.
2. The method of claim 1, wherein at least one of said at least two
different albumin-based drug delivery systems is an albumin-binding
prodrug comprising an albumin-binding group, a drug, and a linker
that can be cleaved hydrolytically, reductively, enzymatically, or
in a pH-dependent manner.
3. The method of claim 1, wherein at least two of said at least two
different albumin-based drug delivery systems are albumin-binding
prodrugs, each comprising an albumin-binding group, a drug, and a
linker that can be cleaved hydrolytically, reductively,
enzymatically, or in a pH-dependent manner.
4. The method of claim 1, wherein at least one of said at least two
different albumin-based drug delivery systems is an albumin-binding
prodrug comprising an albumin-binding group, a drug, and a linker
that can be cleaved hydrolytically, reductively, enzymatically, or
in a pH-dependent manner, and wherein at least one of said two
different albumin-based drug delivery systems is an albumin-drug
nanoparticle.
5. The method of claim 1, wherein the drug contained in each of the
albumin-based drug delivery systems is independently selected from
the group consisting of a cytostatic agent, a cytokine, an
immunosuppressant, an antirheumatic, an antiphlogistic, an
antibiotic, an analgesic, a virostatic, an antimycotic agent, a
transcription factor inhibitor, a cell cycle modulator, an MDR
modulator, a proteasome or protease inhibitor, an apoptosis
modulator, an enzyme inhibitor, an angiogenesis inhibitor, a
hormone or hormone derivative, an antibody or a fragment thereof, a
therapeutically or diagnostically active peptide, a radioactive
substance, a light emitting substance, or a light absorbing
substance.
6. The method of claim 1, wherein the drug contained in each of the
albumin-based drug delivery systems is a cytostatic agent
independently selected from the group consisting of N-nitrosoureas;
the anthracyclines doxorubicin, 2-pyrrollinoanthracycline,
morpholinoanthracycline, diacetatoxyalkylanthracycline,
daunorubicin, epirubicin, idarubicin, mitoxantrone and ametantrone,
and any derivatives thereof; the alkylating agents chlorambucil,
bendamustine, melphalan, and oxazaphosphorines, and any derivatives
thereof; the antimetabolites 5-fluorouracil, 2'-deoxy-5-fluoridine,
cytarabine, cladribine, fludarabine, pentostatine, gemcitabine, and
thioguanine, and any derivatives thereof; the folic acid
antagonists methotrexate, raltitrexed, pemetrexed, and plevitrexed,
and any derivatives thereof; the taxanes paclitaxel and docetaxel,
and any derivatives thereof; the camptothecins topotecan,
irinotecan, SN-38, 10-hydroxycamptothecin, GG211, lurtotecan,
9-aminocamptothecin and camptothecin, and any derivatives thereof;
the Vinca alkaloids vinblastine, vincristine, vindesine, and
vinorelbine, and any derivatives thereof; calicheamicins and and
derivatives thereof; maytansinoids and any derivatives thereof;
auristatins and any derivatives thereof; epothilones and any
derivatives thereof; bleomycin, dactinomycin, plicamycin, miromycin
C and cis-configured platinum(II) complexes, and any derivatives
thereof.
7. The method of claim 2, wherein one or more of the
albumin-binding group(s) bind in situ to cysteine-34 of
albumin.
8. The method of claim 2, wherein one or more of the
albumin-binding group(s) are independently selected from the group
consisting of a maleinimide group, a halogenacetamide group, a
halogenacetate group, a pyridylthio group, a vinyl-carbonyl group,
an aziridin group, a thiol group, a disulfide group, a substituted
or unsubstituted acetylene group, or an N-hydroxysuccinimide ester
group.
9. The method of claim 2, wherein one or more of the
albumin-binding group(s) are independently selected from
phthalocyanines, coumarins, flavonoids, tetracyclines,
naphthalenes, aryl- and heteroarylcarboxylic acids, lipids and
fatty acids, cyclic or linear tetrapyrroles and organometallic
compounds thereof, aromatic acid derivatives substituted with 2 to
5 halogen atoms (CI, Br or I), organic dyes, and the tryptophan and
thyroxine analog compounds, and any derivatives thereof.
10. The method of claim 2, wherein one or more of the cleavable
linker(s) independently comprise a substituted or unsubstituted,
branched-chain or straight-chain aliphatic alkyl group with 1 to 20
carbon atoms, which may comprise one or more oxygen or nitrogen
atoms, and/or a substituted or unsubstituted aryl residue.
11. The method of claim 2, wherein one or more of the cleavable
linkers are enzymatically cleavable and comprise a peptide sequence
selected from Arg, Arg-Arg, Phe-Arg, Phe-Cit, Ile-Pro, Lys,
Lys-Lys, Arg-Lys, Ala-Leu-Ala-Leu (SEQ ID No: 1), Phe-Lys,
Phe-Lys-Ala, Val-Cit, Val-Arg, Ala-Phe-Lys, D-Ala-Phe-Lys, Met,
Met-Met, Phe-Met, Tyr-Met, Ala-Met, Ala-Phe-Met, Phe-Ala-Met,
Ala-Tyr-Met, Phe-Tyr-Met, Ser-Ser-Tyr-Tyr-Ser-Arg (SEQ ID No: 2),
Ser-Ser-Tyr-Tyr-Ser-Leu (SEQ ID No: 3), Arg-Ser-Ser-Tyr-Tyr-Ser-Leu
(SEQ ID No: 4), Phe-Pro-Lys-Phe-Phe-Ser-Arg-Gln (SEQ ID No: 5),
Lys-Pro-Ile-Glu-Phe-Nph-Arg-Leu (SEQ ID No: 6),
Gly-Pro-Leu-Gly-Ile-Ala-Gly-Gln (SEQ ID No: 7),
Gly-Pro-Leu-Gly-Ile-Ala-Gly-Gln (SEQ ID No: 8),
Gly-Pro-Gln-Gly-Ile-Trp-Gly-Gln (SEQ ID No: 9), Gly-Phe-Leu-Gly
(SEQ ID No: 10), or is a p-aminobenzyloxycarbonyl (PABC) linker or
a N-methyl- or symmetric N,N-dimethylethylene linker, or wherein
one or more of the cleavable linker(s) are cleaved upon reduction
and comprise disulfide bonds, or wherein one or more of the
cleavable linker(s) are acid-labile linkers and comprise an
acid-labile bond selected from ester, acetal, ketal, imine,
aconityl, hydrazone, carboxyl-hydrazone and sulfonylhydrazone bonds
and bonds containing a trityl group.
12. The method of claim 1, wherein the at least two different
albumin-based drug delivery systems are each present in separate
containers to be sequentially administered.
13. The method of claim 1, wherein one of said at least two
different albumin-based drug delivery systems is the
6-maleimidocaproyl(hydrazone) derivative of doxorubicin
(DOXO-EMCH), and wherein one of the at least two different
albumin-based drug delivery systems is selected from (i) an
albumin-binding prodrug selected from the methotrexate derivative
EMC-D-Ala-Phe-Lys-Lys(.gamma.-MTX)-OH, wherein
EMC=6-maleimidocaproic acid, (AW054), (ii) an albumin-drug
nanoparticle selected from nab-paclitaxel (ABI-007),
Nab.RTM.-docetaxel (ABI-008) or nab-rapamycin (ABI-010), (iii) an
albumin-binding antibody construct selected from camelid anti-HSA
trivalent nanobodies, and (iv) an albumin fusion protein with
interferons or interleukins selected from albinterferon alfa-2b or
albuleukin.
14. The method of claim 13, wherein one of said at least two
different albumin-based drug delivery systems is the
6-maleimidocaproyl(hydrazone) derivative of doxorubicin
(DOXO-EMCH), and wherein one of the at least two different
albumin-based drug delivery systems is the methotrexate derivative
EMC-D-Ala-Phe-Lys-Lys(.gamma.-MTX)-OH, wherein
EMC=6-maleimidocaproic acid, (AW054).
15. The method of claim 14, wherein the disease to be treated is
cancer.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] The present application claims priority to U.S. application
Ser. No. 14/380,190, filed on Aug. 21, 2014, which is a U.S.
National Stage Application filed under 35 U.S.C. 371 based on
PCT/EP2013/000513 filed on Feb. 21, 2013, which claims the benefit
of EP Application No. 12001136.6 filed on Feb. 21, 2012.
DESCRIPTION
[0002] The present invention relates to a composition comprising at
least two different albumin-based drug delivery systems, as well as
to a pharmaceutical composition comprising said composition.
[0003] Most of the drugs used at present are compounds having low
molecular weights and exhibit, when systemically administered to a
patient, a high plasma clearance or total body clearance.
Furthermore, said low-molecular weight compounds show a high
tendency to penetrate body tissues by diffusion, resulting in a
uniform biodistribution. These are the two main reasons why only
small quantities of the drug reach the site of action and, due to
distribution over healthy tissues of the body, said drugs give rise
to problematic side-effects. These disadvantages are of particular
concern for those drugs having a high cytotoxic potential, such as
cytotoxic agents, immunosuppressive agents or virostatic
agents.
[0004] Several strategies have been pursued for improving the
selectivity of low-molecular weight drugs and thus to increase the
concentration of the active agent in the desired tissue, while the
concentration of the same is decreased in healthy tissues in order
to reduce side-effects.
[0005] Carriers, such as for example albumin, or its drug
conjugates exhibit a markedly long half-life in the systemic
circulation of up to 19 days. Because of an elevated permeability
of vessel walls of for example malignant, infected or inflamed
tissue for macromolecules, the carrier, such as for example serum
albumin, accumulates in the target tissue. Accordingly, such
albumin-based drug delivery systems represent a promising approach
to a more specific and tolerable treatment of such target
tissues.
[0006] In this context, prodrugs have specifically been presented
which bind in situ to e.g. human serum albumin and show improved
properties in contrast to the drug alone. In addition, antibodies,
peptides or synthetic polymers have been investigated as drug
carriers for the development of prodrugs.
[0007] However, although such prodrugs have been shown to allow a
more specific delivery of the active agent to the target tissue, in
most cases it is desirable to further enhance the efficacy and in
vivo tolerability of such agents for an improved treatment, thus
inter alia enabling the effective treatment of diseases that have
been known to be refractory to the drugs currently known in the
art.
[0008] Therefore, the technical problem underlying the present
invention is to provide more effective and/or more tolerable
pharmaceutical compositions that can be used in the treatment of
various diseases.
[0009] The solution to the above technical problem is achieved by
the embodiments characterized in the claims.
[0010] In particular, it has been found that combinations of at
least two albumin-based drug delivery systems surprisingly produce
better therapeutic effects and tolerability in vivo than using
combinations of the low-molecular drugs as such.
[0011] Therefore, in one aspect the present invention relates to a
composition comprising a combination of at least two different
albumin-based drug delivery systems, wherein at least one of the
two different albumin-based drug delivery systems is selected from
albumin-binding prodrugs, albumin drug conjugates, albumin peptide
conjugates, albumin fusion proteins, albumin-binding peptide
conjugates, albumin drug nanoparticles, and albumin-based antibody
constructs.
[0012] According to the present invention, the term "composition"
is not specifically restricted and in general relates to a
combination of at least two components which are present as a
mixture or in separate form and which show an additive or
synergistic effect when used in a combined manner.
[0013] In particular, in the present invention, the term
"composition" includes mixtures of the at least two albumin-based
drug delivery systems, either in a separable or in a non-separable
manner, but also explicitly includes the case where the respective
albumin-based drug delivery systems are present as a combination of
separate components, e.g. in different containers. For Example,
according to the present invention, the composition may be a
mixture of at least two different albumin-based drug delivery
systems e.g. for simultaneous administration of said systems, or
the at least two albumin-based drug delivery systems may be present
in separate containers e.g. for sequential administration, wherein
the combined use of said systems results in an improved
pharmacological effect or improved tolerability, when compared to
the separate use of the single drugs.
[0014] The same principle applies for compositions comprising more
than two components. For example, according to the present
invention, a composition comprising three albumin-based drug
delivery systems may be present in form of a mixture of these three
components, or may be present as a mixture of the first two systems
and, in a separate container, of the third system. As an
alternative, each of the three albumin-based drug delivery systems
may be present in a separate container.
[0015] In the present invention, the expression "albumin-based drug
delivery system" is not specifically restricted and includes those
systems which comprise one or more therapeutically or
diagnostically active agents and use albumin, a fragment or a
derivative thereof, as a protein carrier to improve the delivery of
the drug to the site of action. According to the present invention,
said albumin-based drug delivery systems include those cases where
a therapeutically or diagnostically active agent is physically
and/or chemically bound to albumin, or is capable of binding
thereto. The binding of the drug or its derivative may for example
take place ex vivo to exogenous albumin, i.e. before administering
the albumin-based drug delivery system to the patient, or may take
place in vivo, i.e. after the therapeutically or diagnostically
active agent has been administered to the patient. Examples of
albumin-based drug delivery systems according to the present
invention include those e.g. disclosed in B. Elsadek, F. Kratz:
"Albumin as a drug carrier--new applications on the horizon", J.
Controlled Release, 2011, 157, pp. 4 to 28. Specific examples of
albumin-based drug delivery systems according to the present
invention include Abraxane.TM., which is an albumin-paclitaxel
nanoparticle, and aggregated albumin nano- or microparticles,
drug-albumin conjugates as well as albumin-binding antibody
constructs such as single chain antibodies with high-affinity
albumin-binding peptide domains, camelid anti-HSA trivalent
nanobodies, conjugates of proteins with albumin-binding domain
antibodies and albumin fusion proteins where the protein is an
antibody, an antibody fragment or a cytokine. A further specific
example of the albumin-based drug delivery systems of the present
invention includes albumin-binding prodrugs.
[0016] Herein, the term "different" means that the at least two
different albumin-based drug delivery systems comprised in the
composition of the present invention are not identical and at least
differ in one aspect, such as the molecular structure of at least
one of the components, the ratio and amount of the respective
components, the drugs included therein, the mode of interaction
with the albumin carrier, etc. For example, the composition of the
present invention may contain two albumin-based drug delivery
systems which are identical except that different drugs are
included therein, such as two different cytostatic agents, or a
cytostatic agent and an MDR modulator. According to another
example, the composition as defined above may comprise a
combination of an albumin-binding antibody construct as one
albumin-based drug delivery system, and an albumin-binding prodrug,
e.g. containing a cytostatic agent, as another albumin-based drug
delivery system. According to a further example, the composition as
defined above may comprise a combination of an albumin-drug
nanoparticle as one albumin-based drug delivery system, and an
albumin-binding prodrug, e.g. containing a cytostatic agent, as
another albumin-based drug delivery system.
[0017] According to the present invention, the source of the
albumin present in or used for the albumin-based drug delivery
systems can for example be endogenous or exogenous human serum
albumin or human recombinant albumin or a fragment of albumin.
[0018] One embodiment of the present invention relates to a
composition as defined above, wherein at least one of said at least
two different albumin-based drug delivery systems is an
albumin-binding prodrug comprising an albumin-binding group, a
drug, and a linker that can be cleaved hydrolytically, reductively,
enzymatically, or in a pH-dependent manner.
[0019] Another embodiment of the present invention relates to a
composition as defined above, wherein at least two of said at least
two different albumin-based drug delivery systems are
albumin-binding prodrugs, each comprising an albumin-binding group,
a drug, and a linker that can be cleaved hydrolytically,
reductively, enzymatically, or in a pH-dependent manner.
[0020] According to the present invention, the drug in each of the
albumin-based drug delivery systems may be the same or may be
different. The same applies to compositions which comprise more
than two, i.e. three, four, five or more albumin-based drug
delivery systems. In such cases, each of said albumin-based drug
delivery systems may contain a different drug or the same drug, or
some may contain the same drug while others contain a different
drug. In such a way, it is possible to construct a tailor-made
composition which is individually adjusted to the therapeutic
and/or diagnostic needs.
[0021] The term "prodrug" as used herein relates to any form of a
drug which is administered to an organism, such as a human, in an
inactive or less active form and is converted, e.g. by
metabolization, into the active form. Said conversion of the
prodrug into the active form is not specifically restricted and
includes any chemical and/or physical alteration of the prodrug
which occurs after administration, such as for example release of
an active part of the prodrug at the site of action by hydrolytic,
enzymatic and/or pH-dependent cleavage.
[0022] According to the present invention, there is no specific
restriction as to how the components of the albumin-binding prodrug
of the composition of the present invention, i.e. the
albumin-binding group, the drug, and the cleavable linker are
connected to each other, as long as the drug is bound to the
cleavable linker, either directly or through a spacer group, and
the biological function of the albumin-binding group and the drug
are not negatively affected by the structural setup. The molecular
structure of the albumin-binding prodrug of the composition of the
present invention may, for example, have a linear form or a
branched form or is present in a circular form.
[0023] According to the present invention, there is no specific
restriction concerning the structural setup of the albumin-binding
prodrug of the composition of the present invention, i.e. the way
the constituents of the above-defined prodrug are chemically bonded
together. In particular, the albumin-binding prodrug of the
composition according to the present invention may contain one or
more spacers in any position between the constituents of the
above-defined prodrug, i.e. the albumin-binding group may for
example be bound to the rest of the prodrug through a spacer or, as
another example, the drug may be bound to the cleavable linker
through a spacer. Furthermore, the function of e.g. the cleavable
linker may be incorporated in such a spacer, i.e. a spacer may be
used between the drug and the rest of the albumin-binding prodrug
which can also serve as the cleavable linker. It is also possible
to bind the drug, the cleavable linker, and/or the albumin-binding
group to a central group, which may be linear, branched or cyclic,
such as a peptide, a sugar, a heterocyclic group, or any inorganic
or organic compound suitable to bind one or more of the
constituents of the prodrug. Structural examples of the
albumin-binding prodrugs usable in the composition as defined above
include linear constructs such as (drug)-(cleavable
linker)-(albumin-binding group), (drug)-(spacer)-(cleavable
linker)-(albumin-binding group), (drug)-(cleavable
linker)-(spacer)-(albumin-binding group), or
(drug)-(spacer)-(cleavable linker)-(spacer)-(albumin-binding
group).
[0024] The term "albumin-binding group" as used herein is not
specifically restricted and relates to any functional group which
is capable of binding to albumin by any mechanism, such as covalent
and non-covalent binding. According to the present invention, the
term "albumin-binding group" may include any protein-binding group
which is inter alia or exclusively capable of binding to albumin, a
fragment or a derivative thereof. For example, the albumin-binding
group may be a chemical group capable of binding to an amino, a
hydroxy or a thiol group of albumin. Preferred examples of an
albumin-binding group according to the present invention are a
maleimide group, a halogenacetamide group, a halogenacetate group,
a pyridylthio group, a vinylcarbonyl group, an aziridin group, a
disulfide group, a substituted or unsubstituted acetylene group, a
thiol group, and a hydroxysuccinimide ester group. In a
particularly preferred embodiment of the composition of the present
invention, the albumin-binding group is a maleimide group. The
albumin-binding group may, for example, include functional groups,
such as --COOH or --SO.sub.3H, that can be activated by standard
coupling agents, e.g. dicyclocarbodiimides, acid chlorides, or
peptide coupling reagents (e.g., BOP, HATU, PyBOP).
[0025] Further examples of said albumin-binding group include
compounds or derivatives of phthalocyanines, coumarins, flavonoids,
tetracyclines, naphthalenes, aryl- and heteroarylcarboxylic acids,
lipids and fatty acids, for example long-chain fatty acids such as
C.sub.10-C.sub.20 fatty acids or C.sub.10-C.sub.20 alkyl amines,
cyclic or linear tetrapyrroles and organometallic compounds
thereof, for example porphyrins and protoporphyrins (such as
bilirubin, hematin and derivatives thereof), aromatic acid
derivatives substituted with 2-5 halogen atoms (e.g. F, Cl, Br or
I) such as iophenoxic acid, organic dyes, for example Evans blue
and bromcresol dyes such as bromcresol green and bromcresol purple,
and the tryptophan and thyroxine analog compounds as well as
derivatives of the above-indicated classes of compounds.
Furthermore, according to the present invention, said organic dyes
and their derivatives used as albumin-binding groups can be
chemically modified or derivatized before or after binding to the
therapeutically and/or diagnostically active substance or to the
spacer molecule, whereby however the binding behavior to albumin is
maintained as compared with the unmodified compound. As an example,
a dye used as an albumin-binding group, e.g. an azo dye, may be
derivatized by the above-indicated chemical modification, for
example by reduction of the azo group or by replacement of the azo
group by a C--C single bond or a C--C double bond, in such a way
that it is no longer colored.
[0026] One or several prodrugs can be bound to any suitable
albumin-based carrier such as albumin or fragments and derivatives
thereof. The albumin-based carrier in general contains suitable
functional groups such as hydroxy, amino or thiol groups to bind
the albumin-binding prodrug. If necessary, these can be introduced
in the carrier molecule by chemical modification through techniques
known to those skilled in the art.
[0027] In a preferred embodiment, the albumin-binding group of the
albumin-binding prodrugs usable in the composition according to the
present invention allows said albumin-binding prodrugs to bind in
situ after administration by e.g. injection to serum albumin, to
yield macromolecular prodrugs which carry the drug to the target
site. In a particularly preferred embodiment of the composition of
the present invention, the albumin-binding group of the
above-defined prodrug binds in situ to cysteine-34 of albumin.
According to the present invention, the term "in situ" includes the
binding of the albumin-binding prodrug of the composition according
to the present invention to serum albumin inside the organism to
which said prodrug has been administered.
[0028] The term "cleavable linker" as used herein relates to any
linker which can be cleaved physically or chemically. Examples for
physical cleavage may be cleavage by light, radioactive emission or
heat, while examples for chemical cleavage include cleavage by
redox-reactions, hydrolysis, pH-dependent cleavage or cleavage by
enzymes. Cleavage of the cleavable linker according to the present
invention can be performed in vivo, e.g. in the body of a patient,
e.g. a human patient.
[0029] According to a preferred embodiment of the composition of
the present invention, the cleavable linker comprises one or more
hydrolytically cleavable bonds, the hydrolysis of which releases
the respective drug. Examples for hydrolytically cleavable bonds
are ester bonds or metal-complex bonds, such as are present in
platinum-dicarboxylate complexes, where a diaminediaquoplatinum(II)
complex is liberated.
[0030] In another preferred embodiment of the composition of the
present invention, the cleavable linker may be cleavable by an
enzyme. For example, the cleavable linker of the composition of the
present invention may contain at least one peptide bond which
preferably lies within a cleavable peptide sequence of a protease.
A peptide bond can therefore be implemented by the insertion of a
respective peptide sequence into the cleavable linker. Suitable
enzymes are, for example, proteases and peptidases, e.g. matrix
metalloproteases (MMP), cysteine proteases, serine proteases and
plasmin activators, which are formed or activated in intensified
manner in diseases such as rheumatoid arthritis or cancer, leading
to excessive tissue degradation, inflammations and metastasis.
Preferred examples of proteases according to the present invention
are in particular MMP-2, MMP-3 and MMP-9, cathepsin B, H, L and D,
plasmin, urokinase, and prostate-specific antigen (PSA). Preferred
peptide sequences that are incorporated in the prodrug are: Arg,
Arg-Arg, Phe-Arg, Phe-Cit, Ile-Pro, Lys, Lys-Lys, Arg-Lys,
Ala-Leu-Ala-Leu, Phe-Lys, Phe-Lys-Ala, Val-Cit, Val-Arg,
Ala-Phe-Lys, D-Ala-Phe-Lys, Met, Met-Met, Phe-Met, Tyr-Met,
Ala-Met, Ala-Phe-Met, Phe-Ala-Met, Ala-Tyr-Met, Phe-Tyr-Met,
Ser-Ser-Tyr-Tyr-Ser-Arg, Ser-Ser-Tyr-Tyr-Ser-Leu,
Arg-Ser-Ser-Tyr-Tyr-Ser-Leu, Phe-Pro-Lys-Phe-Phe-Ser-Arg-Gln,
Lys-Pro-Ile-Glu-Phe-Nph-Arg-Leu, Gly-Pro-Leu-Gly-Ile-Ala-Gly-Gln,
Gly-Pro-Leu-Gly-Ile-Ala-Gly-Gln, Gly-Pro-Gln-Gly-Ile-Trp-Gly-Gln,
Gly-Phe-Leu-Gly. In addition, the enzymatically cleavable linker
may contain a self-immolative linker such as a self-immolative
p-aminobenzyloxycarbonyl (PABC) linker or a N-methyl- or symmetric
N,N-dimethylethylene linker.
[0031] In another embodiment, the cleavable linker according to the
present invention contains at least one reductively cleavable bond.
One example of such a reductively cleavable bond is the disulfide
bond. According to a specific example, the disulfide bond both
serves as the albumin-binding group and the cleavable linker.
[0032] In another preferred embodiment of the composition of the
present invention, the cleavable linker according to the present
invention preferably contains at least one acid-labile bond.
Examples of acid-labile bonds are ester, acetal, ketal, imine,
aconityl, hydrazone, acyl hydrazone and sulfonylhydrazone bonds and
bonds containing a trityl group.
[0033] In a further preferred embodiment of the composition of the
present invention, the cleavable linker comprises a substituted or
unsubstituted, branched-chain or straight-chain aliphatic alkyl
group with 1 to 20 carbon atoms, which may comprise one or more
oxygen or nitrogen atoms, and/or a substituted or unsubstituted
aryl residue.
[0034] In a particularly preferred embodiment of the composition of
the present invention, the cleavable linker is an acyl hydrazone
linker.
[0035] The term "drug" as used herein relates to any compound which
brings about a pharmacological effect either by itself or after its
conversion in the organism in question, and thus also includes the
derivatives from these conversions. The pharmacological effect of
the drugs of the composition according to the present invention can
be a single effect only, e.g. a cytostatic effect, or a broad
pharmacological spectrum of actions, such as an immunosuppressive
and antiphlogistic effect at the same time.
[0036] In a preferred embodiment of the composition of the present
invention, the drug contained in each of the albumin-based drug
delivery systems is independently selected from the group
consisting of a cytostatic agent, a cytokine, an immunosuppressant,
an antirheumatic, an antiphlogistic, an antibiotic, an analgesic, a
virostatic, an antimycotic agent, a transcription factor inhibitor,
a cell cycle modulator, an MDR modulator, a proteasome or protease
inhibitor, an apoptosis modulator, an enzyme inhibitor, an
angiogenesis inhibitor, a hormone or hormone derivative, an
antibody or a fragment thereof, a therapeutically or diagnostically
active peptide, a radioactive substance, a light emitting
substance, or a light absorbing substance.
[0037] In a preferred embodiment of the composition of the present
invention, the drug contained in each of the albumin-based drug
delivery systems is a cytostatic agent independently selected from
the group consisting of N-nitrosoureas; the anthracyclines
doxorubicin, 2-pyrrollinoanthracycline, morpholinoanthracycline,
diacetatoxyalkylanthracycline, daunorubicin, epirubicin,
idarubicin, mitoxantrone and ametantrone, and any derivatives
thereof; the alkylating agents chlorambucil, bendamustine,
melphaIan, and oxazaphosphorines, and any derivatives thereof; the
antimetabolites 5-fluorouracil, 2'-deoxy-5-fluoridine, cytarabine,
cladribine, fludarabine, pentostatine, gemcitabine, and
thioguanine, and any derivatives thereof; the folic acid
antagonists methotrexate, raltitrexed, pemetrexed, and plevitrexed,
and any derivatives thereof; the taxanes paclitaxel and docetaxel,
and any derivatives thereof; the camptothecins topotecan,
irinotecan, SN-38, 10-hydroxycamptothecin, GG211, lurtotecan,
9-aminocamptothecin and camptothecin, and any derivatives thereof;
the Vinca alkaloids vinblastine, vincristine, vindesine, and
vinorelbine, and any derivatives thereof; calicheamicins and and
derivatives thereof; maytansinoids and any derivatives thereof;
auristatins and any derivatives thereof; epothilones and any
derivatives thereof; bleomycin, dactinomycin, plicamycin, miromycin
C and cis-configured platinum(II) complexes, or any derivatives
thereof.
[0038] Especially suitable cytokines according to the present
invention are, for example, interleukin-2, interferon .alpha.-2a,
interferon .alpha.-2b, interferon .delta.-1a, interferon .beta.-1b,
interferon .gamma.-1b, tumor necrosis factor, and any derivatives
thereof.
[0039] Especially suitable immunosuppressants according to the
present invention are, for example, cyclosporin A, tacrolimus,
sirolimus, everolimus, mycophenolatmofetil, and any derivatives
thereof.
[0040] Especially suitable antirheumatics according to the present
invention are, for example, methotrexate, leflunomid,
sulfasalazine, chloroquine, and any derivatives thereof.
[0041] Especially suitable antiphlogistic and/or analgesic agents
according to the present invention are, for example, salicylic acid
derivatives such as for example acetylsalicylic acid, and any
derivatives thereof; drug derivatives having an acetic or propionic
acid group such as diclofenac or, respectively, naproxen, and
aminophenol derivatives such as for example paracetamol.
[0042] Especially preferred antibiotics according to the present
invention are, for example, sulfanilamide, sulfacarbamide and
sulfamethoxydiazine, and any derivatives thereof; penicillins, for
example 6-aminopenicillanic acid, penicillin G as well as
penicillin V, and any derivatives thereof; isoxazolylpenicillins
such as oxacillin, cloxacillin and clucloxacillin, and any
derivatives thereof; .alpha.-substituted benzylpenicillins such as
ampicillin, carbenicillin, pivampicillin, amoxicillin, and any
derivatives thereof; acylaminopenicillins, for example mezlocillin,
azlocillin, piperacillin, apalcillin, and any derivatives thereof;
amidinopenicillins, for example mecillinam; atypical .beta.-lactams
such as imipenam and aztreonam; cephalosporins, for example
cephalexin, cefradin, cefaclor, cefadroxil, cefixime, cefpodoxime,
cefazolin, cefazedone, cefuroxime, cefamandole, cefotiam,
cefoxitin, cefotetan, cefmetazole, latamoxef, cefotaxmine,
ceftriaxone, ceftizoxime, cefmonoxime, ceftazidime, cefsulodin and
cefoperazone, and any derivatives thereof; tetracyclines such as
tetracycline, chlorotetracycline, oxytetracycline, demeclocycline,
rolitetracycline, doxycycline, minocycline, and any derivatives
thereof; chloramphenicols such as chloramphenicol and
thiamphenicol, and any derivatives thereof; gyrase inhibitors, for
example nalidixic acid, pipemidic acid, norfloxacin, ofloxacin,
ciprofloxacin and enoxacin, and any derivatives thereof; and
antituberculotics such as isoniazid, and any derivatives
thereof.
[0043] Especially preferred virostatic agents according to the
present invention are, for example, nucleoside analogs such as
acyclovir, ganciclovir, idoxuridine, ribavirin, vidaribine,
zidovudine, didanosine and 2',3'-dideoxycytidine (ddC), and any
derivatives thereof, as well as amantadine.
[0044] Especially suitable antimycotic agents according to the
present invention are, for example, amphotericin B, and any
derivatives thereof.
[0045] Especially preferred MDR modulators according to the present
invention are, for example, verapamil, dihydropyridins, cyclosporin
A and D, tacrolismus, rapamyin, digoxin, digitoxin, quinidin,
lovastatin, atorvastin, analogues of reserpine, trifluoroperazine,
pervilleines A-F, valspodar, dexverapamil, biricodar, bepridil,
erythromycin, levofloxacin, losartan, morphin, rifampin, phenytoin,
colchicin, rhodamin 123, amprenavir, indinavir, nelfinavir,
saqunavir, ritonavir, XR9576, LY335979, OC-144093, R101933,
GF120918, ONT-093, MS-209, S-9788, reversin 205 and 121, or any
related derivative.
[0046] Especially preferred transcription factor inhibitors
according to the present invention are, for example, compounds that
inhibit activation of NF-.kappa.B such as alpha-lipoic acid,
alpha-tocopherol, anetholdithiolthione (ADT), butylated
hydroxyanisole (BHA), cepharanthine, caffeic acid phenethyl ester
(3,4-dihydroxycinnamic acid, CAPE), catechol derivatives,
diethyldithiocarbamate (DDC), diferoxamine, dihydrolipoic acid,
disulfram, dimethyldithiocarbamates (DM DTC), curcumin
(diferuloylmethane), EPC-K1 (phosphodiester compound of vitamin E
and vitamin C), epigallocatechin-3-gallate (EGCG; green tea
polyphenols), ethylene glycol tetraacetic acid (EGTA), glutathione,
L-cysteine, lacidipine, melatonin, N-acetyl-L-cysteine (NAC),
nordihydroguaiaritic acid (NDGA), phenanthrolines,
pyrrolinedithiocarbamate (PDTC), quercetin, tepoxaline
(5-(4-chlorophenyl)-N-hydroxy-(4-methoxyphenyl)-N-methyl-1H-pyrazole-3-pr-
opan-amide), vitamin C, vitamin E derivatives, alpha-torphryl
succinate, alpha-torphryl acetate, PMC
(2,2,5,7,8-pentamethyl-6-hydroxychromane), benzyisocyanate,
resveratol, genistein, lupeol, lycopene, panepoxydone,
epoxyquinomicin C, dehydroxymethylepoxyquinomicin (DHMEQ),
cycloepoxydon, gliotoxin, as well as 1-.kappa.B-alpha
phosphorylation and/or degradation inhibitors such as PS-1,145,
aspirin, salicylic acid, BAY-11-7082
(E3[(4-methylphenyl)-sulfonyl]-2-propenenitrile), BAY-11-7085
(E3[(4-t-butylphenyl)-sulfonyl]-2-propenenitrile), cycloepoxydon;
1-hydroxy-2-hydroxymethyl-3-pent-1-enylbenzene, ibuprofen,
prostaglandin A1, sanguinarine (pseudochelerythrine,
13-methyl-[1,3]-benzodioxolo-[5,6-c]-1,3-dioxolo-4,5
phenanthridinium), sulfasalazine, sulindac, capsaicin
(8-methyl-N-vanillyl-6-nonenamide), emodin
(3-methyl-1,6,8-trihydroxyanthraquinone), erbstatin (tyrosine
kinase inhibitor), estrogen (E2), gliotoxin, genistein,
resiniferatoxin, and miscellaneous inhibitors of NF-.kappa.B such
as beta-amyloid protein, glucocorticoids (dexamethasone,
prednisone, methylprednisolone), leptomycin B (LMB),
o,o'-bismyristoyl thiamine disulfide (BMT), ADP ribosylation
inhibitors e.g., nicotinamide, 3-aminobenzamide, bi-, tri, or
tetracyclic lactames, 1,8-naphtalimide derivatives,
phenanthridin-6-ones, 3,4-dihydro-5-methyl-isoquinolin-1(2H)-one,
benzoxazole-4-carboxamide, 1,6-naphthyridine-5(6H)-ones,
quinazolin[3,4-d]pyrimidin-4(3H)-ones, 1,5-dihydroxyisoquinoline,
2-methyl-quinazolin-4[3H]-ones, 1,11b-dihydro-[2H]benzopyrano
[4,3,2-de]isoquinolin-3-one, atrial natriuretic peptide (ANP),
atrovastatin (HMG-CoA reductase inhibitor), calcitriol
(1a,25-dihydroxyvitamine D3), E3330 (quinone derivative),
herbimycin A, hypericin, hydroquinone (HQ), KT-90 (morphine
synthetic derivatives), mevinolin, 5'-methylthioadenosine (MTA),
pentoxifylline (1-(5'-oxohexyl) 3,7-dimethylxanthine, PTX),
phenyl-N-tert-butylnitrone (PBN), pituitary adenylate
cyclase-activating polypeptide (PACAP), quinadril (ACE inhibitor),
ribavirin, secretory leukocyte protease inhibitor (SLPI), serotonin
derivative (N-(p-coumaroyl) serotonin), silymarin, vasoactive
intestinal peptide (VIP), D609 (phosphatidylcholine-phospholipase C
inhibitor), RO31-8220 (PKC inhibitor), SB203580 (p38 MAPK
inhibitor), triptolide (PG490, extract of Chinese herb), LY294,002,
mesalamine, wortmannin (fungal metabolite), or CHS 828
(N-(6-(p-chlorophenoxy)-hexyl)-N'-cyano-N''-4-pyridylguanidine),
sesquiterpene lactones such as parthenoilde, helenalin,
miller-9E-enolid and budlein A.
[0047] Especially preferred proteasome and protease inhibitors
according to the present invention are, for example, peptide
aldehydes: ALLnL (N-acetyl-leucinyl-leucinyl-norleucynal, MG101),
LLM (N-acetyl-leucinyl-leucinyll-methional), Z-LLnV
(carbobenzoxyl-leucinyl-leucinyl-norvalinal, MG115), Z-LLL
(carbobenzoxyl-leucinyl-leucinyl-leucynal, MG132),
Z-LLL-B(OH).sub.2 (MG-262), boronic acid derivatives, e.g. PS-273,
PS-293, PS-296, PS-303, PS-305, PS-313, PS-321, PS-325, PS-334,
PS-341, PS-364, PS-352, PS-383, lactacystine, beta-lactone, boronic
acid peptide, ubiquitin ligase inhibitors deoxyspergualin, APNE
(N-acetyl-DL-phenylalanine-beta-naphthylester), BTEE (N-benzoyl
L-tyrosine-ethylester), DCIC (3,4-dichloroisocoumarin), DFP
(diisopropyl-uorophosphate), TPCK (N-alpha-tosyl-L-phenylalanine
chloromethyl ketone), TLCK (N-alpha-tosyl-L-lysine chloromethyl
ketone), FK506 (Tacrolimus), Cyclosporin A.
[0048] Especially preferred apoptosis modulators according to the
present invention are, for example, farnesyl transferase
inhibitors, e.g. R115777, SCH66336, BMS214662, Imatinib, 17-AAG,
EGFR inhibitors, e.g., ZD1839, MEK inhibitors, e.g., PD 032590, RAF
inhibitors e.g., BAY43-9006, erlotinib, PKC inhibitors, e.g.
UCN-01, PKC-412, Bryostatin, ISIS-3521, LY333531, safingol,
CGP-41251 (midostaurin), HDAC inhibitors, e.g.,
suberoyl-3-aminopyridineamide hydroxamic acid, lonidamine, apoptin,
survivin, rapamycin, CCI-779, RAD001 (everolimus), PXD101, tyrosine
kinase inhibitors, e.g. Iressa, OSI-774, STI-571, inhibitors of
enzymes in the mitogen-activated protein kinase pathway e.g.,
PD-098059, U-0126.
[0049] Especially preferred cell cycle modulators according to the
present invention are, for example, flavopiridol, bryostain-1,
roscovitine, BMS-387032, perifosine, or lovastatin.
[0050] Especially preferred enzyme inhibitors according to the
present invention are, for example, inhibitors of .gamma.-glutamyl
cystine synthetase e.g., buthione, sulfoxime.
[0051] Especially preferred angiogenesis inhibitors according to
the present invention are, for example, thalidomide, endostatin,
celecoxib, ABT-510, combretastatin A4, dalteparin,
dimethylxanthenone acetic acid, lenalidomide, LY317615
(enzastaurin), PPI-2458, ADH-1 (exherin), AG-013736, AMG-706,
AZD2171, Bay 43-9006 (sorafenib), BMS-582664, CHIR-265, GW786034
(pazopanib), PI-88, PTK787/ZK 222584 (vatalanib), RAD001
(everolimus), SU11248 (sunitinib), suramin, XL184, ZD6474, ATN-161,
or EMD 121974 (cilenigtide).
[0052] Especially preferred hormones or hormone derivatives
according to the present invention are, for example,
aminogluthemid, buserilin, cyproteronacetate, droloxifen,
ethinylestradiol, flutamid, formesta, fosfestrol,
gestonoroncaproate, goserilin, leuprolein, lynestrenol,
medrogeston, medroxyprogesteronacetate, megestrolactetate,
octreotid, tamoxifen, toremifin, triptorelin, anastrazole,
exemestane, or letrozone.
[0053] According to the present invention, there is no specific
limitation regarding the combination of drugs in the composition as
defined above. For example, two or more albumin-based drug delivery
systems which contain drugs with similar pharmacological action,
such as two or more cytostatic agents, two or more antibody
(fragments) or two or more MDR modulators may be combined. On the
other hand, combinations of different types of drugs may be
comprised in the composition as defined above, in case such a
combination is suited to improve the therapeutic effect of the
overall composition.
[0054] In a preferred embodiment of the composition of the present
invention, the drug in each of the said albumin-based drug delivery
systems may be the same or a different specific cytostatic agent,
such as doxorubicin, methotrexate, paclitaxel, docetaxel, or
rapamycin.
[0055] For preparing the albumin-binding prodrug of the composition
of the present invention the drug is bound to a linker through an
acid-sensitive and/or hydrolytically and/or reductively and/or
enzymatically cleavable bond. This derivatization is carried out
with a suitable functional group of the respective drug which is a
HO--, H.sub.2N--, HOOC--, HO.sub.3S--, or carbonyl group. If the
drug does not contain a suitable functional group, then it is
introduced through chemical modification, i.e. the above-mentioned
drugs additionally include all derivatives that possess a HO--,
H.sub.2N--, HOOC--, HO.sub.3S--, and/or carbonyl group.
[0056] A further embodiment of the present invention relates to a
composition as defined above, wherein at least one of said at least
two different albumin-based drug delivery systems is an
albumin-binding prodrug comprising an albumin-binding group, a
drug, and a linker that can be cleaved hydrolytically, reductively,
enzymatically, or in a pH-dependent manner, and wherein at least
one of said two different albumin-based drug delivery systems is an
albumin-drug nanoparticle.
[0057] Specific examples of said albumin-binding nanoparticle
include nab-paclitaxel (ABRAXANE.RTM. ABI-007, Abraxis BioScience),
Nab.RTM.-docetaxel (ABI-008) or Nab.RTM.-rapamycin (ABI-010).
[0058] In a further embodiment of the above-defined composition,
the at least two different albumin-based drug delivery systems are
either present as a mixture, e.g. for simultaneous administration,
or are each present in separate containers e.g. for sequential
administration.
[0059] Herein, the term "simultaneous administration" is not
specifically restricted and means that the individual albumin-based
drug delivery systems are substantially administered at the same
time, e.g. as a mixture or in immediate subsequent sequence.
[0060] Moreover, the term "sequential administration" used herein
is not specifically restricted and means that the individual
albumin-based drug delivery systems are not administered at the
same time but one after the other, or in groups, with a specific
time interval between administrations.
[0061] In a further embodiment, the present invention relates to
the above-defined composition, wherein in the sequential
administration each of the albumin-based drug delivery systems is
administered in specific time interval after the preceding
administration. According to the present invention, said time
interval may be the same or different between the respective
administrations of each of the albumin-based drug delivery systems
and may be selected, for example, from the range of 2 minutes to 96
hours, 1 to 7 days or one, two or three weeks. Generally, according
to the present invention, the time interval between the
administration of the individual albumin-based drug delivery
systems may be in the range of a few minutes to hours, such as in
the range of 2 minutes to 72 hours, 30 minutes to 24 hours, or 1 to
12 hours. Further examples include time intervals in the range of
24 to 96 hours, 12 to 36 hours, 8 to 24 hours, and 6 to 12
hours.
[0062] A specific embodiment of the present invention relates to
the above-defined composition, wherein one of said at least two
different albumin-based drug delivery systems is the
6-maleimidocaproyl(hydrazone) derivative of doxorubicin
(DOXO-EMCH), and wherein one of the at least two different
albumin-based drug delivery systems is selected from (i) an
albumin-binding prodrug, selected from the methotrexate derivative
EMC-D-Ala-Phe-Lys-Lys(.gamma.-MTX)-OH, wherein
EMC=6-maleimidocaproic acid, (AW054), (ii) an albumin-drug
nanoparticle, selected from nab-paclitaxel (ABRAXANE.RTM. ABI-007,
Abraxis BioScience), nab-docetaxel (ABI-008) or nab-rapamycin
(ABI-010), (iii) an albumin-binding antibody construct selected
from camelid anti-HSA trivalent nanobodies, and (iv) an albumin
fusion protein with interferons or interleukins selected from
albinterferon alfa-2b or albuleukin.
[0063] The molar ratio of the albumin-based drug delivery systems
of the composition of the present invention is not particularly
restricted. For example, in case two albumin-based drug delivery
systems are combined in the composition as defined above, the molar
ratio of said systems may be in the range of 1:500 to 500:1, or of
1:100 to 100:1. According to a further example, the molar ratio is
in a range of 1:50 to 50:1, or of 1:20 to 20:1. However, the molar
ratio may also be in the range of 1:5 to 5:1, or 1:1. Similar molar
ratios apply when using three or more albumin-based drug delivery
systems in the composition of the present invention.
[0064] Preferably, the composition of the present invention does
not contain any additional drug, except for the drugs comprised in
each of the at least two different albumin-based drug delivery
systems. That is, according to the present invention, it is
preferred that the above-defined composition comprises the at least
two different albumin-based drug delivery systems, and optionally
one or more adjuvants, such as solvents, diluents, etc., but does
not contain a drug as such which is not part of said albumin-based
drug delivery system.
[0065] The present invention further relates to a kit, comprising
the composition as defined above, and optionally a pharmaceutically
acceptable carrier and/or a pharmaceutically acceptable adjuvant
and/or diluent. In the kit of the present invention, the
albumin-based drug delivery systems may be present as a mixture
e.g. for simultaneous administration, or may be present in separate
containers, e.g. for subsequent administration.
[0066] In another aspect, the present invention relates to a
pharmaceutical composition comprising the composition of the
present invention, and optionally a pharmaceutically acceptable
carrier and/or a pharmaceutically acceptable adjuvant and/or
diluent.
[0067] The kit or the pharmaceutical composition of the present
invention may for example contain solvents and diluents such as a
sodium chloride solution or a solution containing any
pharmaceutically acceptable buffers. Moreover, the pharmaceutical
composition of the present invention may be in any form suitable
for administration to a patient, for example in an injectable form,
as a tablet or a capsule, or as a composition for inhalation.
[0068] According to a further embodiment of the present invention,
the kit or the pharmaceutical composition as defined above are for
use in the treatment of a disease selected from the group
consisting of cancer, autoimmune diseases, acute or chronic
inflammatory diseases, or diseases caused by viruses and/or
microorganisms.
[0069] In a further aspect, the present invention relates to the
use of the composition of the present invention in the manufacture
of a medicament for treating a patient suffering from a disease
selected from the group consisting of cancer, autoimmune diseases,
acute or chronic inflammatory diseases, or diseases caused by
viruses and/or microorganisms.
[0070] A further aspect of the present invention relates to the
composition, the kit, or the pharmaceutical composition as defined
above for use in the treatment of a disease selected from the group
consisting of cancer, autoimmune diseases, acute or chronic
inflammatory diseases, or diseases caused by viruses and/or
microorganisms.
[0071] In a further aspect, the present invention relates to a
method for treating a patient suffering from a disorder or a
disease such as cancer, autoimmune diseases, acute or chronic
inflammatory diseases, or diseases caused by viruses and/or
microorganisms, comprising the step of administering the
composition or the pharmaceutical composition as defined above.
[0072] The Figures show:
[0073] FIGS. 1A and 1B: Chemical structure of the albumin-binding
prodrug DOXO-EMCH (FIG. 1A) which is the
6-maleimidocaproyl(hydrazone) derivative of doxorubicin, and of the
albumin-binding prodrug AW054 (FIG. 1B) which is a methotrexate
derivative.
[0074] FIG. 2: Therapeutic efficacy of DOXO-EMCH (3.times.24
mg/kg), AW054 (3.times.20 mg/kg), and a combination of DOXO-EMCH
(3.times.12 mg/kg) and AW054 (3.times.10 mg/kg) or of doxorubicin
(3.times.6 mg/kg), of methotrexate (3.times.125 mg/kg) or a
combination of doxorubicin (3.times.3 mg/kg doxorubicin) and
methotrexate (3.times.62.5 mg/kg) against the MiaPaCa-2 pancreatic
carcinoma xenograft model (7 animals per group); tumor volumes are
depicted on a logarithmic scale.
[0075] FIG. 3: Body weight curves under therapy with DOXO-EMCH
(3.times.24 mg/kg), AW054 (3.times.20 mg/kg), and a combination of
DOXO-EMCH (3.times.12 mg/kg) and AW054 (3.times.10 mg/kg) or of
doxorubicin (3.times.6 mg/kg), of methotrexate (3.times.125 mg/kg)
or a combination of doxorubicin (3.times.3 mg/kg doxorubicin) and
methotrexate (3.times.62.5 mg/kg) in the MiaPaCa-2 pancreatic
carcinoma xenograft model (7 animals per group) are depicted.
[0076] The compositions of the present invention surprisingly and
advantageously produce an improved efficacy and in vivo
tolerability over the use of the drugs as such, and further provide
an effective treatment of diseases which have previously been
refractory to the therapeutics available in the prior art.
Accordingly, it is advantageously possible to efficiently treat a
great variety of diseases by combining at least two albumin-based
drug delivery systems, such as albumin-binding prodrugs, whereby
the advantageous properties of said drugs can be improved beyond
the properties of the individual drugs alone.
[0077] The present invention will now be further illustrated in the
following example without being limited thereto. Many further
combinations according to the present invention can be realized as
outlined above.
Example
Example: Evaluation of the Antitumor-Efficacy of a Combination of
Two Albumin-Binding Prodrugs
[0078] The antitumor efficacy of a combination of the
6-maleimidocaproyl(hydrazone) derivative of doxorubicin (DOXO-EMCH)
and an albumin-binding prodrug of methotrexate, AW054 (see FIGS. 1A
and B), that is cleaved by two proteases that are over-expressed in
solid tumors, cathepsin B and plasmin was evaluated in the
MiaPaCa-2 pancreatic carcinoma xenograft model.
[0079] The 6-maleimidocaproyl(hydrazone) derivative of doxorubicin
(DOXO-EMCH) (FIG. 1A) is an albumin-binding prodrug of doxorubicin
that binds rapidly to the cysteine-34 position of circulating
albumin after administration and is taken up by solid tumors.
Following tumor uptake, doxorubicin is cleaved in the acidic
environment of tumor tissue, either extra- or intracellularly.
DOXO-EMCH has an maximum tolerated dose (MTD) of 3.times.24 mg/kg
doxorubicin equivalents in nude mice and has shown superior
efficacy over free doxorubicin in several xenograft and orthotopic
tumor models (F. Kratz et al. (2002): Probing the Cysteine-34
Position of Endogenous Serum Albumin with Thiol-binding Doxorubicin
Derivatives: Improved Efficacy of an Acid-sensitive Doxorubicin
Derivative with Specific Albumin-binding Properties Compared to the
Parent Compound, J. Med. Chem. 45, 5523-5533; R. Graeser et al.
(2009): INNO-206, the (6-maleimidocaproyl hydrazone derivative of
doxorubicin); and R. Graeser, N. Esser, H. Unger, I. Fichtner, A.
Zhu, C. Unger, F. Kratz (2010): INNO-206, the (6-maleimidocaproyl
hydrazone derivative of doxorubicin), shows superior antitumor
efficacy compared to doxorubicin in different tumor xenograft
models and in an orthotopic pancreas carcinoma model,
Investigational New Drugs 28, 14-19).
[0080] AW054, i.e. EMC-D-Ala-Phe-Lys-Lys(.gamma.-MTX)-OH
(EMC=6-maleimidocaproic acid), rapidly binds to the cysteine-34
position of circulating albumin after administration to form a
conjugate which is stable in human plasma. Due to two lysine
residues it can be cleaved by cathepsin B and plasmin, two enzymes
which are found in high levels in solid tumors. After binding to
endogenous albumin, AW054 accumulates in tumor tissue and tumor
cells where a methotrexate-lysine derivative is released either
extraor intracellularly by plasmin and cathepsin B, respectively
(A. Warnecke, I. Fichtner, G. Sass, F. Kratz (2007): Synthesis,
cleavage profile, and antitumor efficacy of an albumin-binding
prodrug of methotrexate that is cleaved by plasmin and cathepsin
B., Arch. Pharm., Pharm. Med. Chem. 340, 389-395). AW054 has a MTD
of 3.times.20 mg/kg (methotrexate equivalents) and is superior
compared to methotrexate (MTD 3.times.125 mg/kg) in an ovarian
carcinoma xenograft model (OVCAR3) (A. Warnecke, I. Fichtner, G.
Sass, F. Kratz (2007): Synthesis, cleavage profile, and antitumor
efficacy of an albumin-binding prodrug of methotrexate that is
cleaved by plasmin and cathepsin B., Arch. Pharm., Pharm. Med.
Chem. 340, 389-395).
[0081] The combination of DOXO-EMCH with AW054 was evaluated in
comparison to the respective albumin-binding prodrugs alone as well
as to the conventional drugs doxorubicin and methotrexate and to a
combination of doxorubicin and methotrexate in the MiaPaCa-2
pancreatic carcinoma xenograft model.
[0082] The experiment in the ovarian carcinoma MiaPaCa-2 xenograft
model was carried out as follows:
[0083] In vivo efficacy studies in the MiaPaCa-2 xenograft model
were carried out with doxorubicin, the
6-maleimidocaproyl(hydrazone) derivative of doxorubicin
(DOXO-EMCH), the albumin-binding prodrug of methotrexate AW054, a
combination of the 6-maleimidocaproyl(hydrazone) derivative of
doxorubicin (DOXO-EMCH) and the albumin-binding prodrug of
methotrexate AW054, doxorubicin, methotrexate and a combination of
doxorubicin and methotrexate.
[0084] DOXO-EMCH was dissolved in sterile 10 mM sodium phosphate,
5% D-(+)-glucose (pH 5.8) and the respective dose administered
intravenously within 30 minutes after dissolution. All doses of
DOXO-EMCH and AW054 administered in the studies are stated in
doxorubicin and methotrexate equivalents, respectively.
Adrimedac.RTM. from medac, Germany, was used as the doxorubicin
reference (c=2 mg/mL). Methotrexate (Sodium salt from medac) was
used as a stock solution (12.5 mg methotrexate/mL in 5% D-Glucose
from Braun, sterile). The stock solution AW054 for intravenous
administration was 2.0 mg AW054/mL in 5% D-Glucose from Braun,
sterile.
[0085] For in vivo testing in the MiaPaCa-2 xenograft model, female
NMRI: nu/nu mice (Taconic, Denmark) were used. The mice were held
in individually ventilaged cages (IVC) under sterile and
standardized environmental conditions (25.+-.2.degree. C. room
temperature, 50.+-.10% relative humidity, 12 hour
light-dark-rhythm). They received autoclaved food and bedding
(ssniff, Soest, Germany) and acidified (pH 4.0) drinking water ad
libitum. All animal experiments were performed under the auspices
of the German Animal Protection Law.
[0086] MiaPaCa-2 cells (5.times.10.sup.6 cells/mouse) were
transplanted subcutaneously (s.c.) into the left flank region of
each mouse on day zero. Mice were randomly distributed to the
experimental groups (7 mice per group). Treatment was initiated at
day 10 when the tumors were grown to a size of -60 mm.sup.3. Mice
were treated on a weekly schedule with 3.times.6 mg/kg doxorubicin
(MTD), 2.times.125 mg/kg methotrexate (MTD), 3.times.24 mg/kg
DOXO-EMCH (doxorubicin equivalents) (MTD), 3.times.20 mg/kg AW054
(methotrexate equivalents) (MTD), and a combination of 3.times.12
mg/kg DOXO-EMCH and 3.times.10 mg/kg AW054 or a combination of
3.times.3 mg/kg doxorubicin and 3.times.62.5 mg/kg methotrexate.
All compounds were injected once a week for three weeks (days 10,
17, 24). The injection volume was 0.2 mL/20 g body weight. Tumor
size was measured twice weekly with a caliper-like instrument in
two dimensions. Individual tumor volumes (V) were calculated by the
formula V=(length.times.[width].sup.2)/2 and related to the values
on the first day of treatment (relative tumor volume, RTV). The
experiment was ended on day 43. Statistical analysis was performed
with the U-test (Mann and Whitney) with p<0.05. The body weight
of mice was determined every 3 to 4 days.
[0087] Pancreatic cancer has a poor prognosis: For locally advanced
and for metastatic disease, which collectively represent over 80%
of individuals, median survival is approximately 10 and 6 months,
respectively, when treated with either 5-fluorouracil or
gemcitabine.
[0088] 3.times.6 mg/kg doxorubicin is the maximum tolerated dose
(MTD) in nude mice models. The MTD of methotrexate in nude mice
models is 3.times.125 mg/kg (i.v.). Combination therapy with
doxorubicin and methotrexate was carried out at in a three weekly
schedule (i.v.) at half of their maximum tolerated dose (MTD), i.e.
3.times.3 mg/kg for doxorubicin and 3.times.62.5 mg/kg
methotrexate. Both DOXO-EMCH and AW054 were administered
simultaneously (i.v.) in a three weekly schedule at half of their
maximum tolerated dose (MTD), i.e. 3.times.12 mg/kg for DOXO-EMCH
(doxorubicin equivalents) and 3.times.10 mg/kg (methotrexate
equivalents) and at their respective MTDs, i.e. 3.times.24 mg/kg
for DOXO-EMCH (doxorubicin equivalents) and 3.times.20 mg/kg for
AW054 (methotrexate equivalents) (see the following Table 1).
TABLE-US-00001 TABLE 1 Comparison of doses, mortality, and final
tumor volume of doxorubicin, DOXO-EMCH, and the combination of
DOXO-EMCH and doxorubicin against human MiaPaca-2 cancer xenografts
(median initial tumor volume on day 1 was 0.062 cm.sup.3) Dose
(i.v.) Tumor volume Schedule [mg/kg/ (day 43) Mice Compounds [days]
inject.] toxic death cm.sup.3/d43 7 Glucose- 10, 17, 24 0 0.992
.+-. 0.362- Phosphate Puffer 7 Doxorubicin 10, 17, 24 6 0 0.190
.+-. 0.125* 7 Methotrexate 10, 17, 24 125 0 0.396 .+-. 0.307*
Doxorubicin + 10, 17, 24 3 + 62.5 0 0.199 .+-. 0.071* Methotrexate
7 DOXO-EMCH 10, 17, 24 24 1 (d 39) 0.034 .+-. 0.064* 7 AW054 10,
17, 24 20 0 0.264 .+-. 0.226* 7 DOXO-EMCH + 10, 17, 24 12 + 10 0
0.001 .+-. 0.001* ** AW054 5 .times. 10.sup.6 MiaPaca-2
cells/mouse, s.c. day 0 *Statistically significant to control
(Glucose phosphate buffer treated group ** Statistically
significant to all groups Mann-Whitney test, p < 0.05 NMRI:
nu/nu mice, female
[0089] The efficacy results of the drugs alone and their
combination are shown in FIG. 2
[0090] Compared to the combination of DOXO-EMCH or AW054 alone,
where DOXO-EMCH showed a partial remission (one of the seven mice
died on day 39) and AW054 only showed moderate antitumor effects
over control animals treated with 5% glucose phosphate buffer (see
FIG. 2), the combination of the two albumin-binding prodrugs
produced complete remissions and an acceptable body weight loss of
-13% at the end of the experiment compared to the starting tumor
volume (see FIG. 3).
[0091] With respect to antitumor efficacy, the free drugs
doxorubicin and methotrexate at their optimal and maximum tolerated
doses or as combination of the two drugs only showed moderate
antitumor efficacy up to the end of the experiment (day 43) (see
FIG. 2).
[0092] In summary, treatment with a combination of the two
albumin-binding prodrugs DOXO-EMCH at 3.times.12 mg/kg and AW054 at
3.times.10 mg/kg was the only therapy schedule which produced
complete remissions against subcutaneously growing MiaPaCa-2
pancreatic carcinoma xenografts, this effect being statistically
significant (p<0.05, Mann-Whitney test) compared to all other
groups. Thus, the treatment combining DOXO-EMCH with AW054 has a
distinct advantage over treatment with either DOXO-EMCH or AW054
alone or doxorubicin, methotrexate or a combination thereof with
respect to antitumor efficacy.
Sequence CWU 1
1
1014PRTArtificial SequencePreferred peptide equence incorporated in
the prodrug 1Ala Leu Ala Leu 1 26PRTArtificial SequencePreferred
peptide equence incorporated in the prodrug 2Ser Ser Tyr Tyr Ser
Arg 1 5 36PRTArtificial Sequencecleavable linker 3Ser Ser Tyr Tyr
Ser Leu 1 5 47PRTArtificial SequencePreferred peptide sequence
incorporated in the prodrug 4Arg Ser Ser Tyr Tyr Ser Leu 1 5
58PRTArtificial SequencePreferred peptide sequence incorporated in
the prodrug 5Phe Pro Lys Phe Phe Ser Arg Gln 1 5 68PRTArtificial
SequencePreferred peptide sequence incorporated in the
prodrugnph(6)..(6)x = nitrophenylalanine 6Lys Pro Ile Glu Phe Xaa
Arg Leu 1 5 78PRTArtificial SequencePreferred peptide sequence
incorporated in the prodrug 7Gly Pro Leu Gly Ile Ala Gly Gln 1 5
88PRTArtificial SequencePreferred peptide sequence incorporated in
the prodrug 8Gly Pro Leu Gly Ile Ala Gly Gln 1 5 98PRTArtificial
SequencePreferred peptide sequence incorporated in the prodrug 9Gly
Pro Gln Gly Ile Trp Gly Gln 1 5 104PRTArtificial SequencePreferred
peptide sequence incorporated in the prodrug 10Gly Phe Leu Gly
1
* * * * *