U.S. patent application number 11/805996 was filed with the patent office on 2007-10-11 for compounds for the modulation of the glykolysis-enzyme-and/or of the transaminase-complex.
Invention is credited to Erich Eigenbrodt, Sybille Mazurek, Hans Scheefers.
Application Number | 20070238781 11/805996 |
Document ID | / |
Family ID | 31998414 |
Filed Date | 2007-10-11 |
United States Patent
Application |
20070238781 |
Kind Code |
A1 |
Eigenbrodt; Erich ; et
al. |
October 11, 2007 |
Compounds for the modulation of the glykolysis-enzyme-and/or of the
transaminase-complex
Abstract
The invention relates to compounds for the modulation of the
glycolysis enzyme complex and of the transaminase complex,
pharmaceutical compositions containing such compounds as well as
uses of such compounds for preparing pharmaceutical compositions
for treating various diseases.
Inventors: |
Eigenbrodt; Erich; (Linden,
DE) ; Scheefers; Hans; (Wettenberg-Wissmar, DE)
; Mazurek; Sybille; (Linden, DE) |
Correspondence
Address: |
Mayer Fortkort & Williams, PC
2nd Floor
251 North Avenue West
Westfield
NJ
07090
US
|
Family ID: |
31998414 |
Appl. No.: |
11/805996 |
Filed: |
May 26, 2007 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
10618578 |
Jul 11, 2003 |
7223784 |
|
|
11805996 |
May 26, 2007 |
|
|
|
Current U.S.
Class: |
514/506 ;
435/7.1 |
Current CPC
Class: |
A61P 31/04 20180101;
A61P 35/00 20180101; C07C 239/20 20130101; A61P 19/02 20180101;
C07C 311/24 20130101; A61P 21/04 20180101; A61P 3/10 20180101; C07D
261/18 20130101; A61P 17/06 20180101; A61P 7/00 20180101; A61P 1/04
20180101; A61P 29/00 20180101; A61P 11/06 20180101; A61P 3/04
20180101; A61P 37/02 20180101 |
Class at
Publication: |
514/506 ;
435/007.1 |
International
Class: |
A61K 31/21 20060101
A61K031/21; G01N 33/53 20060101 G01N033/53 |
Foreign Application Data
Date |
Code |
Application Number |
Sep 6, 2002 |
DE |
102 44 080.8 |
Sep 11, 2002 |
DE |
102 42 445.4 |
Sep 23, 2002 |
DE |
102 44 299.1 |
Claims
1. A method for treating cancer comprising administering a
pharmaceutical composition comprising a compound that is mixed with
one or several physiologically well tolerated auxiliary substances
and/or carrier substances and galenically prepared for local, oral,
or systemic administration comprising intravenous administration,
wherein the compound is according to formula I ##STR4## wherein
R1=-H, --CN, --COO+, --COS+, --COOH, --COSH, --COOR1.1, --COSR1.1,
N-phthalimidyl, wherein R1.1=-H, C1-10 alkyl, C1-10 aralkyl or
aryl, wherein R2=-H, C1-C4 alkyl, --OR1.1, -Hal (--F --Cl, --Br,
-J), --NR2.1R2.2, --Am, --O--Am, --S--Am, wherein R3=-H, C1-C4
alkyl, --OR1.1, -Hal (--F --Cl, --Br, -J), --NR2.1R2.2, --Am,
--O--Am, --S--Am, wherein R2.1=-H, C1-10 alkyl, C1-10 aralkyl or
aryl, wherein R2.2=-H, C1-10 alkyl, C1-10 aralkyl or aryl, wherein
R2.1 and R2.2 may be identical or different, wherein n and m may be
identical or different and 0 to 10, wherein o and p may be
identical or different and 0 to 3, wherein o>0, if n and m=0,
wherein R2 and R3 may be identical or different for Cn and/or Cm,
wherein R2 may be identical or different for every Cx=1 . . . n,
wherein R3 may be identical or different for every Cy=1 . . . m,
wherein --Am is an amino acid radical, wherein q and r=0 or 1 and
identical or different, wherein --O.sub.r-- and/or --O.sub.q-- may
also be replaced by --S.sub.r-- or --S.sub.q--, resp., wherein
--NR2.1R2.2 may be replaced by a linear or branched --C1-C20 alkyl,
aralkyl or aryl, wherein a group --CN, --(CO)--CN, --(CO)--O--R1 or
--(CO)--R1 or --C--O--R1 may be replaced by --SO.sub.2--NR2.1R2.2,
or a physiologically well tolerated salt of such a compound.
2. The method according to claim 1, wherein the cancer comprises
tumor cells which overexpress a tumor marker comprising glycolytic
enzyme comprising pyruvate kinase M2 as compared to non-cancerous
cells.
3. The method according to claim 2, wherein the tumor cells
comprise hepatoma cells.
4. The method according to claim 1, wherein the cancer comprises
cancer of the liver.
5. The method according to claim 1, wherein R1=-CN.
6. The method according to claim 1, wherein at least one of the R2
is --Am, wherein --Am preferably represents an amino acid radical
of an essential amino acid, wherein in particular q=0 and r=1 or
q=1 and r=0 or q=1 and r=1, m=1, R3=-H, n=o=p=0, R2.1=R2.2=-H.
7. The method according to claim 1, wherein n=o=p=0, wherein m=0 to
4, wherein R2=R3=-H, or for at least one R2, R2=-Am, wherein
R2.1=R2.2=-H, wherein q=0 and r=1.
8. The method according to claim 1, wherein m=p=0, wherein o=1,
wherein n=0 to 4, wherein R2=H, or for at least one R2, R2=-Am,
wherein R3=-H or -Hal in the case Cx=1, wherein R3=-H for all
Cx=n>1, wherein R2.1=R2.2=-H, wherein q=0 and r=1.
9. The method according to claim 1, wherein m=1 to 4, wherein
n=o=p=0, wherein R2=H, or for at least one R2, R2=-Am, wherein
R3=-H or -Hal in the case Cy=1, wherein R3=-H for all Cy=m>1,
wherein R2.1=R2.2=-H, wherein q=0 and r=1.
10. The method according to claim 1, wherein o=p=1, wherein m=0,
wherein n=0 to 4, wherein R2=R3=-H, or for at least one R2, R2=-Am,
wherein R2.1=R2.2=-H, wherein q=0 and r=1.
11. The method according to claim 1, wherein n=p=0, wherein o=1,
wherein m=0 to 4, wherein R2=R3=-H, or for at least one R2, R2=-Am,
wherein R2.1=R2.2=-H, wherein q=0 and r=1.
12. The method according to claim 1, wherein m=p=0, wherein o=1,
wherein n=1 to 4, wherein R2=R3=-H, or for at least one R2, R2=-Am,
wherein R2.1=R2.2=-H, wherein q=0 and r=1.
13. A method of treating cancer comprising tumor cells which
overexpress a tumor market comprising pyruvate kinase M2 compared
to non-cancerous cells, comprising administering a pharmaceutical
composition comprising a compound according to formula I ##STR5##
wherein R1=-H, --CN, --COO+, --COS+, --COOH, --COSH, --COOR1.1,
--COSR1.1, N-phthalimidyl, wherein R1.1=-H, C1-10 alkyl, C1-10
aralkyl or aryl, wherein R2=-H, C1-C4 alkyl, --OR1.1, -Hal (--F
--Cl, --Br, -J), --NR2.1R2.2, --Am, --O--Am, --S--Am, wherein
R3=-H, C1-C4 alkyl, --OR1.1, -Hal (--F --Cl, --Br, -J),
--NR2.1R2.2, --Am, --O--Am, --S--Am, wherein R2.1=-H, C1-10 alkyl,
C1-10 aralkyl or aryl, wherein R2.2=-H, C1-10 alkyl, C1-10 aralkyl
or aryl, wherein R2.1 and R2.2 may be identical or different,
wherein n and m may be identical or different and 0 to 10, wherein
o and p may be identical or different and 0 to 3, wherein o>0,
if n and m=0, wherein R2 and R3 may be identical or different for
Cn and/or Cm, wherein R2 may be identical or different for every
Cx=1 . . . n, wherein R3 may be identical or different for every
Cy=1 . . . m, wherein --Am is an amino acid radical, wherein q and
r=0 or 1 and identical or different, wherein --O.sub.r-- and/or
--O.sub.q-- may also be replaced by --S.sub.r-- or --S.sub.q--,
resp., wherein --NR2.1R2.2 may be replaced by a linear or branched
--C1-C20 alkyl, aralkyl or aryl, wherein a group --CN, --(CO)--CN,
--(CO)--O--R1 or --(CO)--R1 or --C--O--R1 may be replaced by
--SO.sub.2--NR2.1R2.2, or a physiologically well tolerated salt of
such a compound.
14. The method according to claim 2, wherein the compound inhibits
glycolysis of the tumor cells.
15. The method according to claim 2, wherein the compound inhibits
the activity of the pyruvate kinase M2 of the tumor cells.
16. The method according to claim 1, wherein the cancer comprises
solid tumors.
17. A method for in vitro detection of heart insufficient or
chronic cardiac failure comprising comparing a level of pyruvate
kinase type M2 in a blood plasma sample of a patient with a
predetermined standard level, wherein the level of pyruvate kinase
type M2 in the blood plasma sample is measured by an immunological
assay using monoclonal antibodies targeted to detect pyruvate
kinase type M2.
18. A method for treating one or several diseases from the group
comprising cancer, chronic inflammations, asthma, arthritis,
osteoarthritis, chronic polyarthritis, rheumatic arthritis,
inflammatory bowl disease, degenerative joint diseases, rheumatic
diseases with cartilage disorders, sepsis, autoimmune diseases,
type I diabetes, Hashimoto thyreoiditis, autoimmune
thrombocytopenia, multiple sclerosis, myasthenia gravis,
chronically inflammatory intestinal diseases, Crohn's disease,
uveitis, psoriasis, collagenoses, Goodpasture syndrome, diseases
with disturbed leukocyte adhesion, cachexia, diseases by increased
TNF-alpha concentration, diabetes, adiposity, bacterial infections
including those by antibiotic resistant bacteria comprising
administering a pharmaceutical composition prepared comprising the
compound according to claim 1.
Description
STATEMENT OF RELATED APPLICATION
[0001] This is a continuation of U.S. patent application Ser. No.
10/618,578, filed Jul. 11, 2003, entitled "Compounds For The
Modulation Of The Glykolysis-Enzyme-And/Or Of The
Transaminase-Complex, which is incorporated by reference herein in
its entirety.
FIELD OF THE INVENTION
[0002] The invention relates to compounds for the modulation of the
glycolysis enzyme and/or transaminase complex and thus in
particular to the growth inhibition of cells and/or bacteria,
pharmaceutical compositions containing such compounds as well as
uses of such compounds for preparing pharmaceutical compositions
for treating various diseases.
BACKGROUND OF THE INVENTION
[0003] Cancer is one of the most prevalent causes of death today,
and the number of cancer cases in industrialized countries
continues to grow. This is mainly based on the fact that malignant
tumors are diseases of advanced age and due to successes in
controlling infectious diseases, it is likely that more people will
reach this age.
[0004] In spite of much progress in the diagnostic and therapeutic
fields, the odds for overcoming the most prevalent types of cancer
types are seldom higher than 20%. A cancerous tumor nowadays can be
destroyed or its growth inhibited. A re-conversion of a tumor cell
into a normal cell is, however, not yet possible. Rather, the most
important therapeutic measures, via operation and irradiation,
remove cancer cells from the organism. The presently used
chemotherapeutic agents for cancer, the cytostatics, also lead to
the destruction or damaging of tumor cells only. In most cases, the
effect is so non-specific that simultaneous heavy damage to healthy
cells will occur.
[0005] In general, tumor cells have a metabolism differing from
healthy cells, in particular glycolysis. Thus, a change in the
isoenzyme system involved in glycolysis and a change in the
transport of NADH is typical for tumor cells. Among other effects,
the activity of the enzymes involved in glycolysis is increased.
This permits high reaction rates under the aerobic conditions
typical for tumor cells. For details, reference is made to E.
Eigenbrodt et al., Biochemical and Molecular Aspects of Selected
Cancers, Vol. 2, p. 311 ff (1994).
[0006] Various other diseases mentioned below are either
characterized by an excessive metabolism by glycolysis enzyme
complex and can be treated by the reduction or inhibition
thereof.
PRIOR ART
[0007] From the document E. Eigenbrodt et al., Biochemical and
Molecular Aspects of Selected Cancers, Vol. 2, p. 311 ff (1994), it
is known that glucose analogs are used for inhibiting the
glycolysis. Other approaches known here from are the use of
inhibitors of glycolysis isoenzymes, for instance by suitable
chelation or inhibition of chelations. As a result, the tumor cells
are, in a manner of speaking, starved out. However, a problem with
the above compounds is that many of them are genotoxic and/or not
sufficiently specific for tumor cells.
SUMMARY OF THE INVENTION
[0008] It is the technical object of the present invention to
specify active ingredients that are able to modulate or inhibit the
glycolysis enzyme and transaminase complex, in particular the
proliferation of cancer cells and to thus inhibit the growth of
neoplastic tumors as well as defense over-reactions of the body,
such as septic shock, autoimmune diseases, transplant rejections as
well as acute and chronic inflammatory diseases, and that
simultaneously have only slight to no cytotoxicity at all with
regard to cells which have an intact glycolysis enzyme complex or
other complex structures. In addition, it is intended to inhibit
the growth of unicellular organisms.
BRIEF DESCRIPTION OF THE DRAWINGS
[0009] FIG. 1a is a graphical representation showing the migration
of PGM from the glycolysis enzyme completed into the transaminase
complex.
[0010] FIG. 1b is a graphical representation showing the effect of
aminoacetate (AOA) and hydrozylamine (HA) on the activity of the
cytosolic and mitochondrial isoenzyme of the GOT in vitro.
[0011] FIG. 2a is the structural formula of
5-methyl-N-[4-(trifluoromethyl)phenyl]isoxazole-4-carboxamide.
[0012] FIG. 2b is the structural formula of
(2Z)-2-cyano-3-hydroxy-N-[4-(trifluoromethyl)phenyl]but-2-enamide.
[0013] FIG. 2c is the structural formula of
N-(3,5-dihalogenophenyl)-5-methylisoxazole-4-carboxamide.
[0014] FIG. 2d is the structural formula
(2Z)-2-cyano-N-(3,5-dihalogenophenyl)-3-hydroxybut-2-enamide.
[0015] FIG. 2e is the structural formula of
5-methyl-N-[4-(trihalogenomethoxy)phenyl]isoxazole-4-carboxamide.
[0016] FIG. 2f is the structural formula of
(2Z)-2-cyano-3-hydroxy-N-[4-(trihalogenomethoxy)phenyl]but-2-enamide.
[0017] FIG. 2g is the structural formula of
N-(3-halogenophenyl)-5-methylisoxazole-4-carboxamide.
[0018] FIG. 2h is the structural formula of
(2Z)-2-cyano-N-(3-halogenophenyl)-3-hydroxybut-2-enamide.
[0019] FIG. 3a is the structural formula of methyl
4-cyano-4-oxobutanoate.
[0020] FIG. 3b is the structural formula of ethyl
4-cyano-4-oxobutanoate.
[0021] FIG. 3c is the structural formula of methyl
5-cyano-5-oxopentanoate.
[0022] FIG. 3d is the structural formula of ethyl
5-cyano-5-oxopentanoate.
[0023] FIG. 3e is the structural formula of methyl
5-cyano-3-methyl-5-oxopentanoate.
[0024] FIG. 3f is the structural formula of ethyl
5-cyano-3-methyl-5-oxopentanoate.
[0025] FIG. 3g is the structural formula of methyl
6-cyano-6-oxohexanoate.
[0026] FIG. 3h is the structural formula of ethyl
6-cyano-6-oxohexanoate.
[0027] FIG. 4a is the structural formula of methyl
4-[(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)oxy]-4-oxobutanoate.
[0028] FIG. 4b is the structural formula of methyl
5-[(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)oxy]-5-oxopentanoate.
[0029] FIG. 4c is the structural formula of
2-oxo-5-phenylpentanenitrile.
[0030] FIG. 5a is the structural formula of (aminooxy)acetic
acid.
[0031] FIG. 5b is the structural formula of
(aminooxy)acetaldehyde.
[0032] FIG. 5c is the structural formula of
2-(aminooxy)ethanol.
[0033] FIG. 5d is the structural formula of
O-methylhydroxylamine.
[0034] FIG. 5e is the structural formula of hydroxylamine.
[0035] FIG. 5f is the structural formula of glycol acid.
[0036] FIG. 5g is the structural formula of glycolaldehyde.
[0037] FIG. 5h is the structural formula of ethylene glycol.
[0038] FIG. 5i is the structural formula of acetaldehyde.
[0039] FIG. 6a is the structural formula of
1-(aminooxy)methanesulfonamide.
[0040] FIG. 6b is the structural formula of
1-(aminooxy)-N-phenylmethanesulfonamide.
[0041] FIG. 6c is the structural formula of
(carboxymethoxy)ammonium.
[0042] FIG. 6d is the structural formula of (aminooxy)acetate.
[0043] FIG. 6e is the structural formula of
O-ethylhydroxylamine.
[0044] FIG. 6f is the structural formula of
O-methylhydroxylamin.
DETAILED DESCRIPTION OF THE INVENTION
[0045] For achieving said technical object, the invention teaches a
compound according to formula I ##STR1## wherein R1=-H, --CN,
--COO+, --COS+, --COOH, --COSH, --COOR1.1, --COSR1.1,
N-phthalimidyl, wherein R1.1=-H, C1-10 alkyl, C1-10 aralkyl or
aryl, wherein R2=-H, C1-C4 alkyl, --OR1.1, -Hal (--F, --Cl, --Br,
-J), --NR2.1R2.2, --Am, --O--Am, --S--Am, wherein R3=-H, C1-C4
alkyl, --OR1.1, -Hal (--F --Cl, --Br, -J), --NR2.1R2.2, --Am,
--O--Am, --S--Am, wherein R2.1=-H, C1-10 alkyl, C1-10 aralkyl or
aryl, wherein R2.2=-H, C1-10 alkyl, C1-10 aralkyl or aryl, wherein
R2.1 and R2.2 may be identical or different, wherein n and m may be
identical or different and 0 to 10, wherein o and p may be
identical or different and 0 to 3, wherein o>0, if n and m=0,
wherein R2 and R3 may be identical or different for Cn and/or Cm,
wherein R2 may be identical or different for every Cx=1 . . . n,
wherein R3 may be identical or different for every Cy=1 . . . m,
wherein --Am is an amino acid radical, wherein q and r=0 or 1 and
identical or different, wherein --O.sub.r-- and/or --O.sub.q-- may
also be replaced by --S.sub.r-- or --S.sub.q--, resp., wherein
--NR2.1R2.2 may be replaced by a linear or branched --C1-C20 alkyl,
aralkyl or aryl, wherein a group --CN, --(CO) --CN, --(CO)--O--R1
or --(CO)--R1 or --C--O--R1 may be replaced by
--SO.sub.2--NR2.1R2.2, or a physiologically well tolerated salt of
such a compound.
[0046] An amino acid radical is defined in an amino acid as
follows: NH.sub.2--CHAm--COOH. These are, in particular, amino acid
radicals of the proteinogenic amino acids, especially of the
essential amino acids. For compounds according to the invention
which possess an optical activity (for instance according to
embodiments of claim 3), the various optical variants such as L and
D types are also included. Corresponding considerations apply in
the case of chiral centers.
[0047] Particularly suited are compounds according to the
invention, wherein at least one of the R2 is --Am, wherein --Am
preferably represents an amino acid radical of an essential amino
acid, wherein in particular q=0 and r=1 or q=1 and r=0 or q=1 and
r=1, m=1, R3=-H, n=o=p=0, R2.1=R2.2=-H.
[0048] Further, various specific groups are preferred, namely: i)
wherein n=o=p=0, wherein m=0 to 4, wherein R2=R3=-H, wherein
R2.1=R2.2=-H, wherein q=0 and r=1, ii) wherein m=p=0, wherein o=1,
wherein n=0 to 4, wherein R2=H, wherein R3=-H or -Hal in the case
Cx=1, wherein R3=-H for all Cx=n>1, wherein R2.1=R2.2=-H,
wherein q=0 and r=1, iii) wherein m=1 to 4, wherein n=o=p=0,
wherein R2=H, wherein R3=-H or -Hal in the case Cy=1, wherein R3=-H
for all Cy=m>1, wherein R2.1=R2.2=-H, wherein q=0 and r=1, iv)
wherein o=p=1, wherein m=0, wherein n=0 to 4, wherein R2=R3=-H,
wherein R2.1=R2.2=-H, wherein q=0 and r=1, v) wherein n=p=0,
wherein o=1, wherein m=0 to 4, wherein R2=R3=-H, wherein
R2.1=R2.2=-H, wherein q=0 and r=1, or vi) wherein m=p=0, wherein
o=1, wherein n=1 to 4, wherein R2=R3=-H, wherein R2.1=R2.2=-H,
wherein q=0 and r=1.
[0049] Generally, one R2 may be replaced by --Am.
[0050] Examples for compounds wherein --NR2.1R2.2 is replaced by
--C1-C20 alkyl are: CH.sub.3--O--(CH.sub.2).sub.m--R1,
CH.sub.3--O--CO--(CH.sub.2).sub.m--R1,
CR5R6R7-O--(CH.sub.2).sub.m--R1,
CR5R6R7O--CO--(CH.sub.2).sub.m--R1, wherein R5, R6 and R7 may be
--C1-C10 alkyl, linear or branched, not substituted or substituted.
(CH.sub.2) may of course also be (CR2R3). --O-- or .dbd.O may be
replaced by --S-- or .dbd.S. R1 is as specified above. CR5R6R7 may
in particular be t-butyl.
[0051] Examples for the compounds according to the invention are:
NH.sub.2--O--(CH.sub.2).sub.m--R1,
NH.sub.2--O--(CH.sub.2).sub.n--CO--R1,
NH.sub.2--O--CHHal-(CO).sub.o--R1,
NH.sub.2--O--CHHal-CH.sub.2--(CO).sub.o--R1,
NH.sub.2--O--CHHal-(CH.sub.2).sub.2--(CO).sub.o--R1,
NH.sub.2--O--CHHal-(CH.sub.2).sub.3--(CO).sub.o--R1,
NH.sub.2--O--CHHal-(CH.sub.2).sub.4--(CO).sub.o--R1,
NH.sub.2--O--CO--(CH.sub.2).sub.n--CO--R1,
NH.sub.2--O--CO--(CH.sub.2).sub.n--R1,
NH.sub.2--O--(CH.sub.2).sub.n--CO--R1,
NH.sub.2--O--CO--(CH.sub.2).sub.n--CHNH.sub.2--R1,
NH.sub.2--O--(CH.sub.2).sub.n--CHNH.sub.2--R1, with R1=-CN or
--COOH, m or n=0 to 4, o=0 or 1, wherein --O-- may be replaced by
S.
[0052] Another formula according to the invention is formula II
##STR2## wherein R.sub.a.dbd.--CN, R.sub.b.dbd.--H, .dbd.O, --OH,
--NH.sub.2, R.sub.c.dbd.--NH.sub.2, --O--NH.sub.2,
--O--(C1-10)alkyl, R.sub.d.dbd.--H, -Hal, .dbd.O, --OH, wherein the
case of .dbd.O H the one CH is omitted, wherein w=0 to 10, e.g. 1
to 4.
[0053] Another formula according to the invention is formula III
##STR3## wherein Rp=-R1, --O--R1, --O--(CR2R3).sub.x-R1,
--(CR2R3).sub.x-O--R1, R.sub.q.dbd.--NR2.1R2.2, --O--NR2.1R2.2,
--O--(CR2R3).sub.x-NR2.1R2.2, --(CR2R3).sub.x-O--NR2.1R2.2,
R.sub.r.dbd.--Am, --O--Am, --O--(CR2R3).sub.x-Am,
--(CR2R3).sub.x-O--Am, --R.sub.s.dbd.--H, --C1-C10 alkyl, aryl or
aralkyl, --C1-C10 hydroxyalkyl, aryl or aralkyl, or an ether of
such a hydroxy radical, wherein --O-- may be replaced by --S-- and
x=1 to 10, in particular 1 to 4. R1 is as specified above, in
particular --CN or --COOH. Examples of such compounds are:
NH.sub.2--O--CHAm--R1, NH.sub.2--CHAm--O--R1,
NH.sub.2--O--CHAm--O--R1, NH.sub.2--CHR1-O--Am,
Am--O--CHNH.sub.2--O--R1, NH.sub.2--O--(Am--O--CH--O--R1). On one
side of one --O-- or several --O-- or on both sides of one --O-- or
several --O-- immediately --(CH.sub.2)x-- may be interposed.
[0054] Compounds according to the invention may be present in an
ionized condition in a solution, depending on the pH value (e.g. as
--COO.sup.- in basic condition or --NH.sub.3.sup.+ in acid
condition). Salts, such as hydrochlorides, may also be formed.
[0055] The invention is based on the finding that aside from the
classic metabolic diseases, such as diabetes mellitus, adiposity,
other diseases, such as cancer, autoimmune diseases and rheumatism
are caused by metabolic defects. This explains the strong influence
of diet on these diseases. A directly measurable biochemical
parameter for these metabolic ketoacidoses is the increase of
pyruvate kinase type M2 (M2-PK) growing in the blood of all
diseases above and below. Depending on the respective disease, the
M2-PK detectable in the blood of the patients originates from
different cells: for cancer from tumor cells, for sepsis from
immune cells, for rheumatism from immune and/or synovial cells. In
healthy cells, there are tetrameric forms of the M2-PK in a
high-order cytosolic complex, the glycolysis enzyme complex. By the
over-activation of oncoproteins, there is a migration of the M2-PK
out of the complex and typical changes in the tumor metabolism.
Simultaneously, the phosphoglyceromutase (PGM) leaves the complex
and migrates into another enzyme complex, where the cytosolic
transaminases are associated (see example 2). This complex is
therefore called transaminase complex. The substrate of the PGM,
glycerate-3-P, is the first stage for the synthesis of the amino
acids serine and glycine. Both amino acids are essential for DNA
and polypholipid synthesis. By the migration of the PGM into the
transaminase complex, the synthesis of serine from glutamate and
thus glutaminolysis is activated. The same changes take place in
immune cells, if the immune system fails, such as for instance in
the case of rheumatism, sepsis or polytrauma. The integration of
the metabolism of different cells in multi-cellular organisms takes
place by organ-specific association of the enzymes in the cytosol:
in the muscle, for instance, by association with contraction
proteins. For this reason, the different organs are provided with
respectively specific isoenzymes. The disruption of this order will
necessarily lead to disease. Uni-cellular organisms, such as
bacteria or yeasts which react based on sufficiency of nutrients
with dissipated proliferation, do not have a complex organization
of the cytosol. As a consequence, substances which inhibiting the
failing metabolism of multi-cellular organisms, will also inhibit
the proliferation of such uni-cellular organisms.
[0056] The invention further teaches the use of a compound
according to the invention for preparing pharmaceutical
compositions for treating one or several diseases of the group
comprising "cancer, chronic inflammations, asthma, arthritis,
osteoarthritis, chronic polyarthritis, rheumatic arthritis,
inflammatory bowl disease, degenerative joint diseases, rheumatic
diseases with cartilage disorders, sepsis, autoimmune diseases,
type I diabetes, Hashimoto thyroidotis, autoimmune
thrombocytopenia, multiple sclerosis, myasthenia gravis,
chronically inflammatory intestinal diseases, Crohn's disease,
uveitis, psoriasis, collagenosis, Goodpasture syndrome, diseases
with disturbed leukocyte adhesion, cachexia, diseases by increased
TNF-alpha concentration, diabetes, adiposity, bacterial infections,
in particular with resistant bacteria". The term treatment also
comprises the prophylaxis.
[0057] The invention further teaches a pharmaceutical composition,
wherein a compound according to the invention is mixed with one or
several physiologically well tolerated auxiliary substances and/or
carrier substances and galenically prepared for local or systemic
administration, in particular oral, parenteral, for infusion into a
target organ, for injection (e.g. IV, IM, intracapsular or
intralumbal), for application in tooth pockets (space between tooth
root and gum).
[0058] The invention also teaches the use of a compound according
to the invention for inhibiting in vitro the glycolysis enzyme
complex, in particular of pyruvate kinase, asparaginase, serine
dehydratases, transaminases, desaminases and/or glutaminases. In
particular, transamination, oxidative deamination, hydrolytic
deamination, eliminating deamination, and reductive deamination are
blocked.
[0059] It is understood that if applicable, there may exist
stereoisomers for the compounds according to formula I, such
stereoisomers all being covered by the invention. The term alkyl
comprises linear and branched alkyl groups as well as cycloalkyl,
if applicable also cycloalkyl groups having linear or branched
alkyl substituents. The term aryl also comprises aralkyl groups,
and alkyl substituents may be alkyl or cycloalkyl.
[0060] Surprisingly, it has been found that compounds according to
the invention are able to competitively inhibit the above members
of the glycolysis enzyme complex. The proliferation of cancer cells
in therapeutically relevant concentrations can be inhibited. There
are no cytotoxic effects to be expected in the respective dosage
range. Because of their pharmacological properties, the compounds
according to the invention are also very suitable for the treatment
and prophylaxis of the above-listed diseases. In conjunction with
the indications for the inhibition of inflammations or
anti-rheumatic effects, it is of special relevance that the
substances according to the invention are non-steroidal
substances.
[0061] The inhibition of the glycolysis enzyme and of the
transaminase complex in particular comprises the inhibition of the
metabolic activity and the energy gain from serine, glutamine,
ornithine, proline and arginine or from other amino acids of this
and other families, but also the synthesis of such amino acids used
for energy generation. These are important energy sources in tumor
cells, but also in bacteria and yeast. The cells or bacteria or
yeast are, in a manner of speaking, "starved out." In detail,
substances according to the invention block, for instance, the
following reactions: i) threonine to glycine, ii) threonine to
.alpha.-amino-.beta.-ketobutyrate, iii)
.alpha.-amino-.beta.-ketobutyrate to glycine, iv) serine
pyridoxalphosphate (PLP) Schiff's base to aminoacrylate, in
particular folic acid-dependent serine hydroxymethyltransferase, v)
aminoacrylate to pyruvate (by displacement of the balance of the
natural hydrolysis of the PLP Schiff's base to the Schiff's base),
vi) transamination by means of PLP for the synthesis of an amino
acid from an oxo acid, in particular of the branch-chained
transaminase, the .alpha.-ketoglutarate, oxalacetate,
3-hydropyruvate and glyoxalate transaminase, the glutamate
dehydrogenase. In particular, the formation of pyruvate from amino
acids is inhibited by substances according to the invention.
Important is the release of NH.sub.2--OH or CH3-OH (--H to --C or
--N if applicable replaced by other radicals, for instance alkyl)
by glutaminase, arginase, asparaginase or serine
hydroxymethyltransferase. This will lead to an increased
specificity, since a feature of tumor cells is a high glutaminase
and serine hydroxymethyltransferase activity. NH.sub.2-OH
(hydroxylamine, HA), for instance, can be phosphorylated by the
high pyruvate kinase activities instead of the OH of the phosphate
(e.g. of the ADP). This will lead to a decoupling of the pyruvate
kinase reaction in tumor cells. Therefore, the invention generally
also comprises all natural metabolites of the substances according
to the invention, in particular of aminooxyacetate, i.e. fractions
of these substances.
[0062] In the transaminase complex, in addition to the PGM and
NDPK, the cytosolic isoforms of the glutamate oxalacetate
transaminase (GOT), glutamate pyruvate transaminase (GPT),
glutamate dehydrogenase (GIDH) and malate dehydrogenase (MDH) are
associated. GOT and MDH are components of the malate-aspartate
shuttle, by which the hydrogen produced in the cytosol is
transported into the mitochondria. NAD+ is recycled for the
cytosolic glyceraldehyde 3-phosphate dehydrogenase reaction. The
malate-aspartate shuttle is part of the glutaminolysis. For an
active malate-aspartate shuttle, in addition to GOT, the presence
of the p36-bound form of the MDH is important, as represented in
example 3.
[0063] Various further embodiments of the invention are possible.
For instance, a pharmaceutical composition according to the
invention may comprise several different compounds of the above
definitions. Furthermore, a pharmaceutical composition according to
the invention may, in addition, comprise an active ingredient
different from the compound of formula Ito forms a combination
preparation. Therein, the various employed active ingredients may
be prepared in a single type of administration, i.e. the active
ingredients are mixed in the type of administration. It is,
however, also possible to prepare the various active ingredients in
spatially separated types of administration of identical or
different species.
[0064] As counterions for ionic compounds according to formula I
Na.sup.+, K+, Li+, cyclohexylammonium or basic amino acids (e.g.
lysine, argini, ornithine, glutamine) can be used.
[0065] Drugs prepared by the method according to the invention may
be administered in an oral, intramuscular, periarticular,
intraarticular, intravenous, intraperitoneal, subcutaneous or
rectal manner.
[0066] The invention also relates to methods for preparing drugs
which are characterized by the fact that at least one compound of
formula I is brought into a suitable dosage form by using a
pharmaceutically suitable and physiologically well tolerated
carrier and if applicable further suitable active ingredients, or
any additional or auxiliary substances.
[0067] Suitable solid or liquid dosage forms are for instance
granulates, powders, dragees, tablets, (micro) capsules,
suppositories, syrups, juices, suspensions, emulsions, drops or
injectable solutions as well as preparations with protracted
release of the active ingredient, for the preparation of which,
standard means such as carrier substances, explosion, binding,
coating, swelling, sliding or lubricating agents, flavoring
substances, sweeteners and solution mediators are used.
[0068] Auxiliary substances are, for instance, magnesium carbonate,
titanium dioxide, lactose, mannite and other sugars, talcum, milk
protein, gelatin, starch, cellulose and its derivatives, animal and
plant oils such as cod-liver oil, sunflower, peanut or sesame oil,
polyethylene glycols and solvents, such as sterile water and one or
poly-valent alcohols, e.g. glycerin.
[0069] Preferably the drugs are prepared and administered in dosage
units, each unit containing as an active component a defined dose
of the compound according to formula I of the invention. With solid
dosage units such as tablets, capsules, dragees or suppositories,
this dose may be 1 to 1,000 mg, preferably 50 to 300 mg, and for
injection solutions in an ampule form 0.3 to 300 mg, preferably 10
to 100 mg.
[0070] For treating an adult patient of 50 to 100 kg weight, for
instance 70 kg, for instance daily doses of 20 to 1,000 mg active
ingredient, preferably 100 to 500 mg, are indicated. Under certain
circumstances, higher or lower daily doses may be recommended. The
administration of the daily dose may be a one-time administration
in the form of a single dosage unit or several smaller dosage units
as well as a multi-administration of separated doses in certain
intervals.
[0071] In the following, the invention is explained in more detail
with reference to examples representing embodiments only.
EXAMPLE 1
Quantification of the Effectivity of a Compound According to the
Invention
[0072] Suitable Novikoff hepatoma cells are obtainable from the
tumor bank of the Deutsches Krebsforschungszentrum, Heidelberg
(Cancer Research 1951, 17, 1010). 100,000 cells each are sown out
per 25 cm.sup.2 cultivation bottle. A substance according to the
invention, dissolved in a solvent suitable for use in cell
cultures, for instance water, diluted ethanol, dimethylsulfoxide or
the like, is added in an increasing concentration to the culture
medium, e.g. in a concentration range of 80 .mu.M-5,000 .mu.M or of
100 .mu.M-300 .mu.M. After four days of cultivation, the number of
cells per bottle is counted. In comparison to the control sample
(without addition of a compound according to the invention but
instead with addition of a reference compound), a measurement and
dose dependence of the inhibitive effect of a particular compound
on proliferation can be observed.
EXAMPLE 2
Migration of the PGM
[0073] In FIG. 1a is shown an isoelectric focusing of a tumor cell
extract (MCF-7 cells). It can be seen that PGM leaves the
glycolysis enzyme complex and migrates into a complex associated
with the cytosolic transaminases, the transaminase complex. The
transaminase complex comprises the following: cytosolic glutamate
oxalacetate transaminase (GOT), c-malate dehydrogenase (MDH),
phosphoglyceromutase (PGM). Not shown are: c-glutamate pyruvate
transaminase (GPT), c-glutamate hydroxypyruvate transaminase,
c-alanine hydroxypyruvate transaminase, c-serine hydroxymethyl
transferase and c-glutamate dehydrogenase (GIDH). The PGM and the
nucleotide diphosphate kinase (NDPK) may be associated in the
transaminase as well as in the glycolysis enzyme complex.
EXAMPLE 3
Inhibition of the Malate-Aspartate Shuttle
[0074] In FIG. 1b is shown the effect of aminoacetate (AOA) and
hydroxylamine (HA) on the activity of the cytosolic and
mitochondrial isoenzyme of the GOT in vitro. The isoenzymes of the
GOT were dissociated by an isoelectric focusing. It can be seen
that aminooxyacetate mainly inhibits the cytosolic isoenzyme, and
hydroxylamine inhibits both isoenzymes of the GOT. The inhibition
of the GOT leads to an inhibition of the malate-aspartate shuttle.
As a consequence, NAD cannot be recycled, and the glycolysis is
inhibited at the stage of the GAPDH.
[0075] The following explanations are independent from the above
examples. The invention further teaches the use of
N-(4'-trifluoromethylphenyl)-5-methylisoxazole-4-carboxamide
(C.sub.12H.sub.9F.sub.3N.sub.2O.sub.2; MW 270.2, see also FIG. 2a)
and/or its natural active metabolites A 77 1726 according to FIG.
2b for preparing a pharmaceutical composition for treating tumor
diseases, in particular solid tumors. The benzene ring may,
alternatively to the shown substitution with --CF3, generally be
singly, doubly, triply, quadruply or quintuply substituted with
-Chal3 or --O-Chal3 or -Hal at an arbitrary position. The
pharmaceutical composition according to the invention is
particularly suited for treating large tumors, i.e. beginning from
0.1 to 1 cm.sup.3 tumor size. A pharmaceutical composition
according to the invention is, for instance, prepared for oral
administration, for instance with the following auxiliary and
carrier substances: colloidal SiO.sub.2, crospovidone,
hydroxypropylmethyl cellulose, lactose monohydrate, magnesium
stearate, polyethylene glycol, povidone, starch, talcum, TiO.sub.2
and/or yellow iron oxide. The dosage is 1 to 50 mg per day,
preferably 10 to 30 mg. It may be recommended to administer in a
therapy initially at a starting dose of 20 to 500 mg, in particular
50 to 150 mg, for the first 1 to 10 days, in particular the first 1
to 3 days. In another embodiment of the invention, the substance
mentioned above is combined with one or several sugar phosphates,
for instance fructose-1,6-biphosphate, glycerate-2,3-biphosphate,
glycerate-3-phosphate, ribose-1,5-biphosphate,
ribulose-1,5-biphosphate, and the combination of substances in a
dosage form, for instance a tablet, may be mixed. It is however
also possible to provide the components separately in identical or
different dosage forms. The sugar phosphate may be administered in
a dosage of 20 to 5,000 mg per day, for instance 100 to 500 mg.
[0076] These variants of the invention surprisingly lead to an
inhibition of the growth of tumor cells and tumor tissue, since
these substances or the metabolite can bind to the pyruvate kinase
and inhibit or reverse the failure of energy metabolism in tumor
cells. This results in the beneficial effect that these substances
specifically influence the metabolism of tumor cells and they do
not or to a lesser extent influence the metabolism of normal cells,
having only slight side effects, if at all.
[0077] The effectiveness of these substances is surprising because
the known effect of a pyrimidine synthesis inhibitor relates to a
completely different effective mechanism, and the empirical
observation of an anti-proliferative effect is substantially
directed toward immune cells and cells related to inflammatory
diseases.
[0078] Of special importance is a combination of one or several of
the active ingredients mentioned on the previous page with one or
several of the active ingredients mentioned further above or
aminooxyacetate (AOA, NH2-O--CH2-COOH, salts or esters thereof, for
instance C1-C10 alkyl or hydoxyalkyl esters). For instance, AOA is
particularly effective for small tumors (<0.1 to 1 cm.sup.3) or
prevents the development thereof, in particular, the development of
metastases, whereas compounds of the formulas 10 or 11, if
applicable in combination with sugar phosphate, is effective for
large tumors. The reason for this is due to the different
metabolisms of small and large tumors. The above explanations for
combinations apply in an analogous manner.
[0079] Substances according to the invention can further be used
for preparing a pharmaceutical composition for treating heart
insufficiency or the chronic cardiac failure (CCF). These are the
variants defined by the New York Heart Association (NYHA)
Classification or grades from NYHA I to NYHA IV. All of these
diseases are marked by an acute and/or chronic failure of the heart
muscle to provide for blood circulation or the transportation
capacity required for the metabolism of the organism under load or
at rest. The reasons are: insufficient glycolysis by glucose
deficiency in the heart muscle and/or its insufficient oxygen
supply and complex coronary inflammation processes (activation of
cells of the immune system and complement). This aspect of the
invention is based on the finding that the substances according to
the invention provide modulation of alternative energy-generating
biochemical processes, and thus make it possible to produce
replacement pathways for the above insufficiently operating
processes. For instance, this is achieved by activation of the
serinolysis and glutaminolysis or to displacement by the substances
according to the invention, the existing dynamic balance between
glycolysis on the one hand and glutaminolysis on the other hand in
favor of the glycolysis, under simultaneous administration of
oxygen (increase in the oxygen partial pressure in the blood by,
for instance, breathing). In this context, the administration of
anti-inflammatory substances according to the invention can prevent
imminent highly dangerous acidosis (by lactate production).
Compared to prior art measures, such as administration of ACE
inhibitors, diuretics, digitalis, positive inotropic substances or
isosorbide dinitrate, the substances according to the invention
directly influence energy metabolism, and the latter is improved.
Side effects are as a consequence, comparatively weak.
[0080] In this context, it has been found by the invention that at
least in the cases of the NYHA grade II to IV, the concentration of
tumor M2-PK (=M2-PK dimeric in contrast to standard M2-PK being
tetrameric) in cells and/or the blood increases, and can be
detected in a routine manner, as an alternative to other methods
typically utilized to date. Therefore the invention further teaches
the use of a tumor M2-PK detecting test system for preparing a
diagnostic substance for the in vitro diagnosis of a heart
insufficiency, in particular, the grade or the inflammatory
processes connected therewith. If increased M2-PK values (sick
collective) are found in the blood plasma of a patient compared to
standard values (defined maximum limits; normal collective), this
is indicative of the existence of a heart insufficiency and/or for
inflammatory processes correlated therewith, or at least identifies
a risk of heart insufficiency. Such a blood plasma analysis can
easily and quickly be performed. Compared thereto, previous
standard methods (gold standard, blood gas analysis) are not
suitable for routine tests and are expensive. For this aspect of
the invention, any known test systems can be used which detect
tumor M2-PK, e.g. immunological test systems with antibodies. In
particular, per se known test systems can be used which detect
tumor M2-PK as a tumor metabolism marker, for instance monoclonal
antibodies specific herefor.
[0081] Various substances which can be used according to the
invention are shown in the further figures, FIGS. 2a-2h, 3a-3h,
4a-4c, 5a-5i, and 6a-6f. In particular, the essential variation
possibilities are represented in an exemplary manner, the
permutations which are easily deduced not being shown for the sake
of simplicity. The invention finally also comprises all natural
metabolites of the described substances.
* * * * *