U.S. patent application number 09/402455 was filed with the patent office on 2002-04-25 for benzyloxy prodigiosine compounds.
Invention is credited to BARGIOTTI, ALBERTO, COLOTTA, FRANCESCO, D'ALESSIO, ROBERTO, FERRARI, MARIO, ISETTA, ANNA MARIA, TIBOLLA, MARCELLINO, VANOTTI, ERMES.
Application Number | 20020049248 09/402455 |
Document ID | / |
Family ID | 10826711 |
Filed Date | 2002-04-25 |
United States Patent
Application |
20020049248 |
Kind Code |
A1 |
D'ALESSIO, ROBERTO ; et
al. |
April 25, 2002 |
BENZYLOXY PRODIGIOSINE COMPOUNDS
Abstract
A compound which is a
2-(1H-pyrrol-2-yl)-5[(2H-pyrrol-2-ylidene)methyl]-1H- -pyrrole of
formula (I), wherein R1 is hydrogen or C.sub.1-C.sub.5 alkyl; R2 is
a C.sub.1-C.sub.5 alkyl; or a pharmaceutically acceptable salt
thereof, is useful as an immunomodulating agent and for treating
adult-T-cell leukemia-lymphoma. 1
Inventors: |
D'ALESSIO, ROBERTO;
(CINISELLO BALSAMO, IT) ; VANOTTI, ERMES; (MILAN,
IT) ; BARGIOTTI, ALBERTO; (MILAN, IT) ;
TIBOLLA, MARCELLINO; (SENAGO, IT) ; FERRARI,
MARIO; (MILAN, IT) ; ISETTA, ANNA MARIA; (RHO,
IT) ; COLOTTA, FRANCESCO; (MILAN, IT) |
Correspondence
Address: |
OBLON SPIVAK MCCLELLAND
MAIER & NEUSTADT
1755 JEFFERSON DAVIS HIGHWAY
FOURTH FLOOR
ARLINGTON
VA
22202
|
Family ID: |
10826711 |
Appl. No.: |
09/402455 |
Filed: |
October 7, 1999 |
PCT Filed: |
January 21, 1999 |
PCT NO: |
PCT/EP99/00415 |
Current U.S.
Class: |
514/422 ;
548/518 |
Current CPC
Class: |
A61P 35/02 20180101;
A61P 43/00 20180101; C07D 207/44 20130101; A61P 37/02 20180101 |
Class at
Publication: |
514/422 ;
548/518 |
International
Class: |
A61K 031/40; C07D
405/00; C07D 207/00; C07D 295/00 |
Foreign Application Data
Date |
Code |
Application Number |
Feb 9, 1998 |
GB |
9802745.1 |
Claims
1. A compound which is a prodigiosine of formula (I) 11wherein R1
is hydrogen or C.sub.1-C.sub.5 alkyl; and R2 is C.sub.1-C.sub.5
alkyl; or a pharmaceutically acceptable salt thereof.
2. A compound according to claim 1, wherein R1 is hydrogen or
methyl; and R2 is a C.sub.1-C.sub.5 alkyl.
3. A compound selected from:
4-benzyloxy-5-[(5-methyl-2H-pyrrol-2-ylidene)-
methyl]-2,2'-bi-1H-pyrrole;
4-benzyloxy-5-[(5-ethyl-2H-pyrrol-2-ylidene)me-
thyl]-2,2'-bi-1H-pyrrole;
4-benzyloxy-5-[(5-propyl-2H-pyrrol-2-ylidene)met-
hyl]-2,2'-bi-1H-pyrrole;
4-benzyloxy-5-[(5-isopropyl-2H-pyrrol-2-ylidene)m-
ethyl]-2,2'-bi-1H-pyrrole;
4-benzyloxy-5-[(5-butyl-2H-pyrrol-2-ylidene)met-
hyl]-2,2'-bi-1H-pyrrole;
4-benzyloxy-5-[(5-pentyl-2H-pyrrol-2-ylidene)meth-
yl]-2,2'-bi-1H-pyrrole;
5'-methyl-4-benzyloxy-5-[(5-methyl-2H-pyrrol-2-yli-
dene)methyl]-2,2'-bi-1H-pyrrole;
5'-methyl-4-benzyloxy-5-[(5-ethyl-2H-pyrr-
ol-2-ylidene)methyl]-2,2'-bi-1H-pyrrole; and the pharmaceutically
acceptable salts thereof.
4. A compound according to claim 3 which is a hydrochloride,
hydrobromide or methanesulphonate salt.
5. A compound as defined in claim 1 for use in a method of
treatment of the human or animal body by therapy.
6. A compound as claimed in claim 5, for use as an immunomodulating
agent or in the treatment of adult-T-cell leukemia-lymphoma.
7. A pharmaceutical composition comprising a pharmaceutically
acceptable carrier and/or diluent and, as an active principle, a
compound as defined in claim 1.
8. A product containing a compound as defined in claim 1 and an
additional drug as a combined preparation for simultaneous,
separate or sequential use in immunosuppressive therapy in
mammals.
9. A pharmaceutical composition for use in immunosuppressive
therapy in a mammal, including a human, comprising: (a) an
immunosuppressive agent in a pharmaceutically acceptable carrier
and/or excipient, and (b) at least one compound as defined in claim
1 in a pharmaceutically acceptable carrier and/or excipient, in
amounts to produce a superadditive immunosuppressant effect.
10. A pharmaceutical composition having activity against
adult-T-cell leukemia-lymphoma, which comprises a compound as
defined in claim 1 as an active ingredient, and a pharmaceutically
acceptable carrier or diluent.
11. A product containing a compound as defined in claim 1, and an
anti-tumor agent as a combined preparation for simultaneous,
separate or sequential use in adult-T-cell leukemia-lymphoma
therapy.
12. A process for the preparation of a compound as defined in claim
1, which process comprises reacting a compound of formula (II)
12wherein R2 is as defined in claim 1 and X is a leaving group,
with a compound of formula (III) 13 wherein R1 is as defined in
claim 1 and R7 is hydrogen or a lower alkyl chain; and, if desired,
salifying a compound of formula (I) and/or, if desired, converting
a salt of a compound of formula (I) into a free compound and/or, if
desired, separating a mixture of isomers of a compound of formula
(I) into the single isomers.
13. A method of treating a mammal, including a human, in need of an
immunomodulating agent, which method comprises administering to
said mammal an effective amount of a compound as defined in claim
1.
14. A method of treating a mammal, including a human, suffering
from adult-T-cell leukemia-lymphoma, which method comprises
administering to said mammal a therapeutically effective amount of
a compound as defined in claim 1.
15. An immunosuppressive therapy method for use in a mammal,
including a human, in need thereof, which method comprises
administering to the mammal (a) an immunosuppressive agent and (b)
at least one compound as defined in claim 1, in amounts effective
to produce a superadditive immunosuppressive effect.
16. A method for lowering the side effects caused by
immunosuppressive therapy with an immunosuppressive agent in a
mammal, including a human, in need thereof, which method comprises
administering to said mammal a combination preparation comprising
(a) said immunosuppressive agent and (b) at least one compound as
defined in claim 1 in a quantity effective to produce a
superadditive immunosuppressive effect.
17. A combined method of treatment of adult-T-cell
leukemia-lymphoma in a mammal, including a human, in need thereof,
which method comprises administering thereto a compound as defined
in claim 1, and an anti-tumor agent, in amounts and close enough
together in time sufficient to produce a therapeutically useful
effect.
Description
[0001] The present invention relates to novel benzyloxy
prodigiosine compounds, to a process for their preparation, to
pharmaceutical compositions containing them and to their use as
therapeutic agents, in particular as immunomodulating agents.
[0002] International application WO 95/17381 discloses
2,2'-bi-1H-pyrrole compounds endowed with high in vitro
immunosuppressive activity. However such compounds, similarly to
other known prodigiosine compounds, are characterized by high
lipophylicity, low aqueous solubility and consequently low
bioavailability.
[0003] Moreover, the task to combine in the same prodigiosine
molecule a high immunosuppressive activity and adequate
hydrosolubility cannot be achieved by merely introducing
hydrophilic groups into the structure of in vitro active
immunosuppressants, as in most cases this strategy results in a
significant loss of immunosuppressive activity. In fact, as known
in the art, the therapeutic efficacy of all drugs is thoroughly
influenced by different parameters that can affect their
bioavailability. Object of the present invention is to provide
novel prodigiosine compounds endowed with improved
bioavailability.
[0004] The present invention is based on the discovery that a
sub-genus of compounds disclosed in WO 95/17381, besides possessing
good bioavailability, have in vivo high immunosuppressive
activity.
[0005] Object of the present invention are novel compounds which
are 2-(1H-pyrrol-2-yl)-5[(2H-pyrrol-2-ylidene)methyl]-1H-pyrroles
i.e. prodigiosine compounds, having the following formula (I) 2
[0006] wherein
[0007] R1 is hydrogen or a C.sub.1-C.sub.5 alkyl; and
[0008] R2 is C.sub.1-C.sub.5 alkyl;
[0009] or pharmaceutically acceptable salts thereof.
[0010] The present invention includes within its scope all possible
isomers, stereoisomers and optical isomers and their mixtures, and
the metabolites and the metabolic precursors or bioprecursors of
the compounds of formula (I).
[0011] The compounds of the invention can be represented also by
the following tautomeric formula (Ia) 3
[0012] wherein R1 and R2 are as defined above.
[0013] Accordingly, the chemical compounds provided by the present
invention are named throughout the description of the invention
according to the chemical nomenclature provided for the compounds
of either formula (I) or (Ia), on the basis of the structural
evidence validated by people skilled in the art.
[0014] The alkyl groups may be branched or straight chain
groups.
[0015] R1 as alkyl group is preferably methyl or ethyl.
[0016] R2 is preferably a methyl, ethyl, propyl, isopropyl, butyl
or pentyl group.
[0017] Examples of pharmaceutically acceptable salts of the
compounds of the invention are the salts with inorganic, e.g.
hydrochloric, hydrobromic and sulphuric acids and with organic
acids, e.g. citric, tartaric, maleic, malic, fumaric,
methanesulphonic and ethanesulphonic acids.
[0018] As stated above, the present invention also includes within
its scope pharmaceutically acceptable bio-precursors (otherwise
known as pro-drugs) of the compounds of formula (I), i.e. compounds
which have a different formula to formula (I) above, but which
nevertheless upon administration to a human being are converted
directly or indirectly in vivo into a compound of formula (I).
[0019] Preferred compounds of the invention are those wherein, in
formula (I),
[0020] R1 is hydrogen or methyl;
[0021] R2 is a C.sub.1-C.sub.5 alkyl.
[0022] Examples of particularly preferred compounds of the
invention are:
[0023]
4-benzyloxy-5-[(5-methyl-2H-pyrrol-2-ylidene)methyl]-2,2'-bi-1H-pyr-
role;
[0024]
4-benzyloxy-5-[(5-ethyl-2H-pyrrol-2-ylidene)methyl]-2,2'-bi-1H-pyrr-
ole;
[0025]
4-benzyloxy-5-[(5-propyl-2H-pyrrol-2-ylidene)methyl]-2,2'-bi-1H-pyr-
role;
[0026]
4-benzyloxy-5-[(5-isopropyl-2H-pyrrol-2-ylidene)methyl]-2,2'-bi-1H--
pyrrole;
[0027]
4-benzyloxy-5-[(5-butyl-2H-pyrrol-2-ylidene)methyl]-2,2'-bi-1H-pyrr-
ole;
[0028]
4-benzyloxy-5-[(5-pentyl-2H-pyrrol-2-ylidene)methyl]-2,2'-bi-1H-pyr-
role;
[0029]
5'-methyl-4-benzyloxy-5-[(5-methyl-2H-pyrrol-2-ylidene)methyl]-2,2'-
-bi-1H-pyrrole;
[0030]
5'-methyl-4-benzyloxy-5-[(5-ethyl-2H-pyrrol-2-ylidene)methyl]-2,2'--
bi-1H-pyrrole;
[0031] and the pharmaceutically acceptable salts thereof, in
particular the hydrochlorides, hydrobromides and
methanesulphonates.
[0032] A further object of the present invention is to provide a
compound of formula (I), as defined above, for use in a method of
treatment of the human or animal body by therapy, in particular as
an immunomodulating agent, especially as an immunosuppressive
agent.
[0033] Object of the present invention is also to provide a
pharmaceutical composition comprising a pharmaceutically acceptable
carrier and/or diluent and as an active principle a compound of
formula (I), as herein defined, or a pharmaceutically acceptable
salt thereof.
[0034] The present invention also provides a method of treating a
mammal, including humans, in need of an immunomodulating agent,
said method comprising administering to said mammal an effective
amount of a compound of formula (I), as herein defined, or a
pharmaceutically acceptable salt thereof.
[0035] The compounds of formula (I) and the pharmaceutically
acceptable salts thereof can be obtained by an analogy process.
[0036] According to a preferred embodiment of the invention a
compound of formula (I) and the salts thereof can be prepared by a
process comprising reacting a compound of formula (II) 4
[0037] wherein
[0038] R2 is as defined above and X is a leaving group, with a
compound of formula (III) 5
[0039] wherein
[0040] R1 is as defined above and R7 is hydrogen or a lower alkyl
chain;
[0041] and, if desired, salifying a compound of formula (I) and/or,
if desired, converting a salt of a compound of formula (I) into a
free compound and/or, if desired, separating a mixture of isomers
of a compound of formula (I) into the single isomers.
[0042] When R7 is a lower alkyl chain, it is preferably a
C.sub.1-C.sub.4 alkyl chain, for instance methyl, ethyl or
isopropyl.
[0043] In a compound of general formula (II), the leaving group X
can be for instance a trifluoromethane-sulphonate group or a
halogen such as chlorine, bromine or iodine.
[0044] The reaction between a compound of formula (II) and a
compound of formula (III) may be carried out in a suitable organic
solvent such as tetrahydrofurane, dioxane, dimethoxyethane, DMF,
toluene, methanol, ethanol, water or mixtures thereof, in the
presence of a suitable palladium (0) catalyst, in the presence of a
basic agent, such as K.sub.2CO.sub.3, Na.sub.2CO.sub.3,
NaHCO.sub.3, K.sub.3PO.sub.4, NaOAc, KOH, NaOH, Ba(OH).sub.2,
EtONa, Bu.sub.4NF, Et.sub.3N, at a temperature varying between
about 60.degree. C. and about 120.degree. C., for a time of about 1
hour to about 3 days.
[0045] A wide range of palladium (0) catalysts can be used such as
for instance Pd(PPh.sub.3).sub.4, PdCl.sub.2(PPh.sub.3).sub.2,
Pd(OAc).sub.2 plus PPh.sub.3 or other ligands as described for
example in Chem. Rev. 95, 2457 (1995).
[0046] Optionally, salt such as LiCl, LiBr, KCl, KBr can be added
to stabilize the catalyst.
[0047] According to a preferred embodiment of the invention, when
in a compound of formula (II) the leaving group X is
trifluoromethanesulfonate- , a preferred catalyst is
Pd(PPh.sub.3).sub.4 in the presence of sodium or potassium
carbonate, and the reaction can be performed in dioxane or toluene,
at a temperature varying between about 65.degree. C. and about
90.degree. C., for a time from about 5 hours to about 24 hours.
[0048] Also the optional salification of a compound of formula (I)
as well as the conversion of a salt into the free compound and the
separation of a mixture of isomers into the single isomers may be
carried out by conventional methods. For example, the separation of
optical isomers may be carried out by salification with an
optically active base or acid and by subsequent fractional
crystallization of the diastereoisomeric salts, followed by
recovering of the optically active isomeric acids or, respectively,
bases.
[0049] A compound of formula (II) can be obtained from a compound
of formula (IV) 6
[0050] wherein
[0051] R2 is as defined above, by means of an opportune reagent
such as for instance trifluoromethane-sulfonic anhydride or a
halogenating agent such as POCl.sub.3, POBr.sub.3,
POCl(OEt).sub.2/TMSI in an inert organic solvent such as
dichloromethane, dichloroethane, acetonitrile, optionally in the
presence of an organic base such as Et.sub.3N or pyridine, at a
temperature varying between about -20.degree. C. and about
50.degree. C. (as described for example in PCT/EP 97/00368).
[0052] The compounds of formula (III) can be prepared as described
in published procedures, as for instance in J. Org. Chem. 46, 157
(1981) and Synthesis, 613 (1991).
[0053] The compounds of formula (IV) can be prepared reacting a
compound of formula (V) 7
[0054] wherein
[0055] R2 is as defined above,
[0056] with 4-benzyloxy-3-pyrrolin-2-one (VI) 8
[0057] The condensation between a compound of formula (V) and the
compound of formula (VI) can be performed by acidic or basic
catalysis, in a solvent such as water, methanol, ethanol, dioxane,
THF, DMF, DMSO or mixtures thereof, at a temperature varying from
about 25.degree. C. to about 120.degree. C., in a time ranging from
about 1 hour to about 24.
[0058] A acidic catalyst can be e.g. an inorganic acid such as HCl,
HBr, H.sub.2SO.sub.4, H.sub.2NO.sub.3 or an organic acid such as,
for instance, p-toluensulphonic acid, methansulphonic acid,
trifluoromethan-sulphonic acid or trifluoroacetic acid.
[0059] As well, a basic catalyst can be e.g. an inorganic base such
as NaOH, KOH, K.sub.2CO.sub.3, Ba(OH).sub.2, NaH or an organic base
such as, for instance, t-BuOk, MeLi, BuLi, LDA.
[0060] The compounds of formula (V) can be prepared, for example,
by Vilsmeier formylation of the compounds of formula (VII) 9
[0061] wherein R2 is as defined above, according to well known
chemical procedures.
[0062] The compounds of formula (VII) are known compounds or may be
prepared using mere variations of published procedures, for example
those reported in the following chemical literature: Tetrahedron
32, 1851 (1976); J.Org.Chem. 53, 1410 (1988); J.Org.Chem. 28, 857
(1963); J.Am.Chem.Soc. 84, 4655 (1962); Ann. 450, 181 (1926); Ber.
99, 1414 (1966).
[0063] The compound of formula (VI) are commercially available or
can be synthesized as described for example in Synthesis, 391
(1992) and Tetrahedron Letters 25, 1871 (1984).
[0064] When in the compounds of formula (I), and in the
intermediate products thereof, groups are present which need to be
protected before submitting them to the here above illustrated
reactions, they may be protected before the reactions take place
and then deprotected, according to well known methods in organic
chemistry.
[0065] The compounds of formulae (I) and the pharmaceutically
acceptable salts thereof are herein defined as the "compounds of
the present invention", the "compounds of the invention" and/or the
"active principles of the pharmaceutical compositions of the
invention".
PHARMACOLOGY
[0066] The compounds of the invention have immunomodulating, in
particular immunosuppressive, in vivo activity as found for example
in the "Delayed-Type Hypersensitivity" assay and a remarkable
bioavailability after oral administration as compared to products
described in the prior art.
Comparative In Vivo Activity Evaluation by DTH Assay
[0067] The immunosuppressive activity of the compounds of the
invention was evaluated in vivo by DTH (Delayed-Type
Hypersensitivity) assay. According to the test, sheep red blood
cells (SRBC) (1.times.10.sup.5 cells) suspended in 500 mcL saline,
were injected i.v. into the tail vein of female C57 Bl/6 mice (8-9
week old). Five days later 1.times.10.sup.8 SRBC suspended in 50
mcL saline were injected into the left hind footpad. The increase
in footpad thickness was measured with a dial micrometer 24 h after
challenge. The test compounds were given daily for six days at
different doses starting on the day of priming. Activity was
expressed as ED30 (dose able to reduce by 30% the thickness
increase compared to controls).
In Vivo Bioavailability Evaluation
[0068] Aim of the study is to determine the pharmacokinetics and
the oral bioavailability of the instant prodigiosine compounds in
rats.
[0069] Species/strain/sex: rat/Lewis/male
[0070] No. formulations: 1 oral; 1 intravenous
[0071] No. animals/formulation: 3+2 controls (+an ulterior rat
treated with the iv formulation); total 9
[0072] Dosages: iv: 1 mg salt/kg; oral: 10 mg salt/kg
[0073] Vehicles: iv: a solution at the conc. of 5 mg/ml in PEG
400/Tween 80 (6:1 v:v) was prepared, then diluted with dextrose at
the final concentration of 0.5 mg/ml; oral: a solution at the
concentration of 5 mg/ml in Cremophor ELP/EtOH abs (6.5:3.5 v:v)
was prepared, then diluted with saline at the final concentration
of 1 mg/ml
[0074] Experimental: Three cannulated rats/formulation were
treated. One rat/formulation was only treated with the vehicle, as
basal sample. The intravenous administration was given into the
caudal vein as bolus; the oral administration by gastric gavage as
solution. Blood was withdrawn from the superior vena cava from each
rat and collected into heparinized tubes at the following times: 2'
(only for the iv route), 5', 15', 30'; 1, 2, 4, 6, 8, 24 and 32 h
post-dosing. Plasma was immediately obtained by centrifugation
(10000 rpm for 3 min) and stored in labelled tubes at -30.degree.
C. till analysis.
[0075] Analytical assay: the extraction of the compounds was
performed by protein precipitation by adding 100 mcl of
acetonitrile to 25 mcl of plasma. The concentrations of the
compounds (as free bases) in plasma were determined by LC-MS
method. Column: APEX CN RP 5 mc, 10.times.4 mm (Jones
chromatography); mobil phase: 70% acetonitrile/30% 1 mM ammonium
formate+0.01% triethylamine adjusted to pH 2.0 with formic acid;
flow rate: 1 ml/min; injection volume: 10 mcL; oven temperature:
45.degree. C.; MS detection by APCI using MRM positive ion; MRM
transition: 330 m/z>239 m/z, retention time: 1.4 min; LLOQ: 5.03
ng/ml; ULOQ: 10060 ng/ml. Bioavailability is expressed in following
Table 1 as F%. ClogP is a lypophylic index, related to
octanol/water partition coefficient.
[0076] Following Table 1 shows the results obtained for two
representative compounds of the invention in comparison with a
closely-related prior art compound.
1TABLE 1 10 DTH ED30 (mg/kg) Compound R1 R2 ClogP iv os F % PNU
168727 H --CH.sub.3 3.98 1.7 5.3 35 PNU 169417 H --C.sub.5H.sub.11
5.64 1.1 4.4 18 PNU 156804 H --C.sub.11H.sub.23 8.02 0.44 18.7
7
[0077] In Table 1:
[0078] PNU 168727 means
4-benzyloxy-5-[(5-methyl-2H-pyrrol-2-ylidene)methy-
l]-2,2'-bi-1H-pyrrole, hydrochloride;
[0079] PNU 169417 means
4-benzyloxy-5-[(5-pentyl-2H-pyrrol-2-ylidene)methy-
l]-2,2'-bi-1H-pyrrole, hydrochloride;
[0080] PNU 156804 means
4-benzyloxy-5-[(5-undecyl-2H-pyrrol-2-ylidene)meth-
yl]-2,2'-bi-1H-pyrrole, hydrochloride.
[0081] PNU 156804 is the most active compound disclosed by WO
95/17381, as shown on page 32 of such prior art reference where it
is coded as FCE 29002.
[0082] In view of their valuable biological properties the
compounds of the invention can therefore be useful in mammals,
including humans, as immunosuppressive agents for the prevention
and treatment of rejection phenomena associated with tissue and
organ transplantations, graft-versus-host diseases and autoimmune
diseases. A mammal, comprising humans, in need of an
immunomodulating agent, in particular of an immunosuppressive
agent, can therefore be treated by a method comprising the
administration thereto of a therapeutically effective amount of a
compound of the invention or a pharmaceutically acceptable salt
thereof. The condition of the human or animal patient can thus be
improved.
[0083] Preferred cases of organ and tissue transplants which can be
successfully treated by the compounds of the invention, here above
described, are, for example, the cases of heart, kidney, bone
marrow, lung, liver, and multiple organ transplantations.
[0084] Preferred cases of autoimmune diseases which can be
successfully treated by the compounds of the invention, here above
described, are for example, the cases of rheumatoid arthritis,
systemic lupus erythematosus, juvenile diabetes, autoimmune
haemolytic anaemia, miastenia gravis, multiple sclerosis,
psoriasis, ulcerative colitis, idiopathic thrombocytopenic purpura,
active chronic hepatitis, glomerulonephritis, idiopathic
leucopenia, primary biliary cirrhosis, thyroiditis, thyrotoxicosis,
dermatomyositis, discoid lupus erythematosus, psoriatic arthritis,
regional enteritis, nephrotic syndrome, lupus nephritis, lupoid
hepatitis, Sjogren's syndrome, Goodpasture's syndrome, Wegener's
granulomatosis, scleroderma, Sezary's disease, uveitis and mumps
orchitis. Typically rheumatoid arthritis, systemic lupus
erythematosus, juvenile diabetes, miastenia gravis, multiple
sclerosis and psoriasis.
[0085] The therapeutic regimen for the different clinical syndromes
must be adapted to the type of pathology taking into account, as
usual, also the route of administration, the form in which the
compound is administered and the age, weight and conditions of the
subject involved.
[0086] The oral route is employed, in general, for all conditions
requiring such compounds. Preference is given to intravenous
injection or infusion for the acute treatments.
[0087] For maintenance regimens the oral or parenteral, e.g.
intramuscular or subcutaneous, route is preferred.
[0088] For these purposes the compounds of the invention, can be
administered orally at doses ranging e.g. from about 0.5 to about
10 mg/kg of body weight per day in adult humans.
[0089] Doses of active compounds ranging e.g. from about 0.25 to
about 5 mg/kg of body weight per day can be used for the parenteral
administration and for intravenous injection or infusion in adult
humans. Of course, these dosage regimens may be adjusted to provide
the optimal therapeutic response. The nature of the pharmaceutical
compositions containing the compounds of this invention in
association with pharmaceutically acceptable carriers or diluents
will, of course, depend upon the desired route of
administration.
[0090] The compositions may be formulated in the conventional
manner with the usual ingredients. For example, the compounds of
the invention, may be administered in the form of aqueous or oily
solutions or suspensions, tablets, pills, gelatine capsules,
syrups, drops or suppositories.
[0091] Thus, for oral administration, the pharmaceutical
compositions, containing the compounds of this invention, are
preferably tablets, pills or gelatine capsules which contain the
active substance together with diluents, such as lactose, dextrose,
sucrose, mannitol, sorbitol, cellulose; lubricants, for instance
silica, talc, stearic acid, magnesium or calcium stearate, and/or
polyethylene glycols; or they may also contain binders, such as
starches, gelatine, methylcellulose, carboxymethylcellulose,
gum-arabic, tragacanth, polyvinylpyrrolidone; disaggregating
agents, such as starches, alginic acid, alginates, sodium starch
glycolate; effervescing mixture; dyestuffs; sweeteners; wetting
agents, such as lecithin, polysorbates, laurylsulphates and in
general, non-toxic and pharmacologically inactive substances used
in pharmaceutical formulations.
[0092] Said pharmaceutical preparations may be manufactured in
known manner, for example by means of mixing, granulating,
tabletting, sugar-coating, or film-coating processes.
[0093] The liquid dispersions for oral administration may be e.g.
syrups, emulsions and suspensions.
[0094] The syrups may contain as carrier, for example, saccharose
or saccharose with glycerine and/or mannitol and/or sorbitol.
[0095] The suspensions and the emulsions may contain as carrier,
for example, a natural gum, agar, sodium alginate, pectin,
methylcellulose, carboxymethylcellulose, or polyvinyl alcohol.
[0096] The suspensions or solutions for intramuscular injections
may contain together with the active compound a pharmaceutically
acceptable carrier, e.g. sterile water, olive oil, ethyl oleate,
glycols, e.g. propylene glycol, and if desired, a suitable amount
of lidocaine hydrochloride.
[0097] The solutions for intravenous injections or infusions may
contain as carrier, for example, sterile water or preferably they
may be in the form of sterile aqueous isotonic saline
solutions.
[0098] The suppositories may contain together with the active
compound a pharmaceutically acceptable carrier, e.g. cocoa butter,
polyethylene glycol, a polyoxyethylene sorbitan fatty acid ester
surfactant or lecithin.
[0099] The present invention also provides products containing a
compound of formula (I), or a pharmaceutically acceptable salt
thereof, and an additional drug as a combined preparation for
simultaneous, separate or sequential use in immunosuppressive
therapy in mammals.
[0100] Such additional drug can be for instance a corticosteroid,
an immunosuppressive or an anti-tumor agent, or mixtures of two or
more of them.
[0101] The term "antitumor agent" is meant to comprise both a
single anti-tumor drug and "cocktails", i.e. a mixture of such
drugs according to clinical practice.
[0102] Examples of anti-tumor agents that can be formulated in
immunosuppressive therapy with a compound of formula (I), include
methotrexate and cyclophosphamide and mixtures thereof.
[0103] The term "immunosuppressive agent" is meant to comprise both
a single immunosuppressive drug and "cocktails", i.e. a mixture of
such drugs according to clinical practice.
[0104] Examples of immunosuppressive agents that can be formulated
with a compound of formula (I), include for instance one of the
following:
[0105] cyclosporin A or cyclosporin C, a non-polar cyclic
oligopeptide; FK506, a fungal macrolide immunosuppressive;
azathioprine, or
6-[(1-Methyl-4-nitro-1H-imidazol-5-yl)-thio]1H-purine;
methotrexate; rapamycin, a fungal macrolide immunosuppressive;
mycophenolate mofetil, or
6-(4-hydroxy-6-methoxy-7-methyl-3-oxo-1,3-dihydroisobenzofuran-5-yl)-4-
-methyl-4-(E)-hexenoic acid 2-(4-morpholinyl)-ethyl ester; an
immunosuppressive glucocorticoid, such as prednisone or
dexamethasone; and/or polyclonal, such as a anti-human thymocite
antibody or a monoclonal such as a anti-human CD3 antibody; or a
mixture of two or more thereof.
[0106] It has to be noted that co-administration of an
immunosuppressive agent, as defined above, and at least one
benzyloxy prodigiosine compound of formula (I), or a
pharmaceutically acceptable salt thereof, as herein defined,
produces a potentiated immunosuppressive activity in synergistic
way, thus giving a superadditive immunosuppressive effect, i.e.
effect which is greater than the sum of the actions of the
individual components. A person skilled in the art will appreciate
that such superadditive immunosuppressive effect allows
administration of lower dosage levels of immunosuppressive agents,
thus lowering the side effects caused by commonly used
immunosuppressant agents.
[0107] Accordingly, the present invention also provides a
pharmaceutical composition for use in immunosuppressive therapy in
mammals, including humans, comprising:
[0108] (a) an immunosuppressive agent in a pharmaceutically
acceptable carrier and/or excipient, and
[0109] (b) at least one compound of formula (I), as defined above,
or a pharmaceutically acceptable salt thereof in a pharmaceutically
acceptable carrier and/or excipient, in amounts to produce a
superadditive immunosuppressant effect.
[0110] A further aspect of the present invention is an
immunosuppressive therapy method for use in mammals, including
humans, in need thereof, the method comprising administering to
said mammal (a) an immunosuppressive agent and (b) at least one
compound of formula (I), as defined above, or a pharmaceutically
acceptable salt thereof, in amounts effective to produce a
superadditive immunosuppressive effect.
[0111] In view of the combined therapeutic effect obtainable by
such combined preparation, lower doses of immunosuppressive agents
can be used.
[0112] Accordingly, the invention also provides a method for
lowering the side effects, such as nephrotoxicity and/or
hepatotoxicity, caused by immunosuppressive therapy with an
immunosuppressive agent in mammals, including humans, in need
thereof, the method comprising administering to said mammal a
combination preparation comprising (a) said immunosuppressive agent
and (b) at least one compound of formula (I), as defined above, or
a pharmaceutically acceptable salt thereof, in a quantity effective
to produce a superadditive immunosuppressive effect.
[0113] In the combined preparations, pharmaceutical compositions
and method of treatment according to the present invention only one
compound of formula (I), as defined above, or a pharmaceutically
acceptable salt therapy, is preferably used. The combination
preparation according to the invention can also include combination
packs or compositions in which the constituents are placed side by
side and can therefore be administered simultaneously, separately
or sequentially to one and the same mammal, including humans.
[0114] The benzyloxy prodigiosine compounds of formula (I) and the
pharmaceutically acceptable salts thereof have also been found to
be active in treating adult-T-cell leukemia-lymphoma, in particular
brought on by infection with HTLV-I in mammals, including humans.
Such therapeutic activity of the compounds of the invention is
proven for instance by the fact that they have been found to be
active in inhibiting selectively the IL-2 induced activation and
expansion of murine and human T-cells, showing thus a
pharmacological profile consistent with the therapy of the IL-2
dependent ATL.
Inhibition of IL-2 Proliferation Induced Activity
[0115] The Th.sub.2 murine cells D10-G4.1 (ATCC TIB 224) are IL-2
dependent for their growth. They are cultured in complete RPMI 1640
medium enriched with rhIL-2 (6 ng/ml) and ConA (6 ng/ml).
[0116] For testing the inhibitory effects of the compounds of the
invention on IL-2 activity, D 10 cells are washed twice with
complete medium, resuspended at 10.sup.5 cells/ml in the same
medium and triplicately distributed (10.sup.4 cells/well) in flat
bottomed 96 well plates. 50 ml of rhIL-2 and 50 ml of the test
compound at different concentrations are simultaneously added to
the cells. The cultures are then incubated at 37.degree. C. in a
humidified 5% CO.sub.2 incubator for 48 h, the last 18 h in the
presence of 0.2 .quadrature.Ci of .sup.3H-TdR.
[0117] Uptake of 3H-TdR in the cells (cpm) is taken as a measure of
cell proliferation.
[0118] For instance for the representative compound of the
invention
4-benzyloxy-5-[(5-methyl-2H-pyrrol-2-ylidene)methyl]-2,2'-bi-1H-pyrrole
(internal code PNU 168727) the following activity data were
obtained.
2 % inhibition Compound ng/ml .sup.3HTdR uptake* vs vehicle PNU
168727 30 52 (8) 100 10 8100 (34) 87 3 54594 (424) 11 1 58245
(1133) 5 vehicle -- 61231 (1193) *mean cpm from triplicate wells
(SE)
[0119] In treating a adult-T-cell leukemia-lymphoma one or more
compound of formula (I), as defined above, can be administered
alone or in association with an anti-tumor agent. Preferably a
single compound of formula (I) is used.
[0120] Accordingly, the present invention provides a compound of
formula (I), as herein defined, or a pharmaceutically acceptable
salt thereof for use in treating adult-T-cell
leukemia-lymphoma.
[0121] A further object of the present invention is a method of
treating mammals, including humans, suffering from adult-T-cell
leukemia-lymphoma, said method comprising administering a
therapeutically effective amount of a compound of formula (I), as
herein defined, or a pharmaceutically acceptable salt thereof.
[0122] Object of the present invention is also to provide a
pharmaceutical composition having activity against adult-T-cell
leukemia-lymphoma comprising a compound of formula (I), as herein
defined, or a pharmaceutically acceptable salt thereof, as an
active ingredient, and a pharmaceutically acceptable carrier or
diluent.
[0123] A further object of the present invention is to provide a
combined method of treatment of adult-T-cell leukemia-lymphoma in
mammals, including humans, in need thereof, said method comprising
administering thereto a compound of formula (I), or a
pharmaceutically acceptable salt thereof, and an anti-tumor agent,
in amounts and close enough together in time sufficient to produce
a therapeutically useful effect.
[0124] The present invention also provides a product containing a
compound of formula (I), or a pharmaceutically acceptable salt
thereof, and an anti-tumor agent as a combined preparation for
simultaneous, separate or sequential use in adult-T-cell
leukemia-lymphoma therapy.
[0125] The term "anti-tumor agent" is meant to comprise both a
single anti-neoplastic agent and "cocktails", i.e. a mixture of
such drugs according to clinical practice.
[0126] An anti-neoplastic agent in treating adult-T-cell leukemia
lymphoma can be for example an agent selected from the group
consisting of an antineoplastic vinca alkaloid, an antineoplastic
antibiotic, an antineoplastic antimetabolite, an antineoplastic
platinum coordination complex, an antineoplastic taxane compound,
an antineoplastic ceramide compound, an antineoplastic distamycin
compound, an antineoplastic epidophyllotoxin compound and an
antineoplastic topoisomerase I inhibitor.
[0127] Examples of specific antineoplastic agents, according to the
invention, which are administered with a compound of formula (I),
are: vincristine, vinblastine, etoposide, tallimustine-amidoxime,
3-(1-methyl-4-(1-methyl-4-(1-methyl-4-(4,N,N-bis(2-chloroethyl)aminobenze-
ne-1-carboxamido)pyrrole-2-carboxamido)pyrrole-2-carboxamido)pyrrole-2-car-
boxamido)proprionamidoxime,
(2S-RR-4E)-1,3-dihydroxy-2-tetradecanoylamido-- 4-octadecene,
paclitaxel, docetaxel, 7-epitaxol, 7-epitaxotere, epirubicin,
doxorubicin, cyclophosphamide, idarubicin, 4'-iodoxorubicin,
daunorubicin, actinomicin D, bleomycin, plycamicin, mitomycin,
camptothecin, 9-aminocamptothecin, camptothecin 11 (CPT 11),
topotecan, metotrexate, cytarabine, azauridine, azarabine,
fluorodeoxyuridine, deoxycoformycin, mercaptopurine, cisplatin and
carboplatin.
[0128] In particular they are epirubicin, doxorubicin,
cyclophosphamide, 9-aminocamptothecin and camptothecin 11.
[0129] The dosage of a compound of the invention to be administered
to a patient suffering from adult-T-cell leukemia-lymphoma, in
particular brought on by infection with HTLV-I, will vary with the
precise nature of the conditions being treated and the recipient of
the treatment.
[0130] A therapeutically effective dosage of the compounds of
formula (I), for example the compound
4-benzyloxy-5-[(5-methyl-2H-pyrrol-2-ylidene]-2,- 2'-bi-1H-pyrrole
hydrochloride (PNU 168727), is in the range of about 0.03 to about
1.5 mg/kg, preferably about 0.06 mg/kg to about 0.7 mg/kg when
given i.v. whereas the dose of the same compound for oral
administration in adult humans is in general from about 0.3
mg/kg/day to about 15 mg/kg/day.
[0131] The dosage of a compound of formula (I) and of an antitumor
agent, in case of combined therapy, to be used is, of course,
dependent on various factors such as the organism to be treated
(e.g., human or animal, age, weight, general state of health), the
severity of the symptoms, the disorder to the accompanying
treatment with other pharmaceuticals, or the frequency of the
treatment. The dosages are in general administered several times
per day and preferably once to three times per day. The effective
amounts of the antitumor agent are in general those commonly used
in therapy, as known to those skilled in the art. However, the
amounts of the individual active compounds should be within the
range given above, e.g. within the tolerable, efficacious dosage
range for the organism to be treated.
[0132] The oral route is employed, in general, for all conditions
requiring the compounds of the invention. Preference is given to
intravenous injection or infusion for the acute treatments. For
maintenance regimens the oral or parenteral, e.g. intramuscular or
subcutaneous, route is preferred.
[0133] The nature of the pharmaceutical preparations and
compositions according to the invention will of course depend upon
the desired route of administration.
[0134] The compositions may be formulated in the conventional
manner with the usual ingredients, for instance as described above.
The following examples illustrate but do not limit the
invention.
EXAMPLE 1
Compound (IV)
[0135] To a solution of 2-formyl-5-methylpyrrole (4 g; 36.65 mmols)
and 4-benzyloxy-3-pyrrolin-2-one (8.24 g; 43.78 mmols) in DMSO (65
ml) 2N sodium hydroxyde (45 ml) was added under nitrogen atmosphere
and the mixture was stirred at 60.degree. C. for 8 hours. After
dilution with water (200 ml) the yellow suspension was filtered.
The crude material was taken up in ethyl acetate, stirred and
filtered to give
4-benzyloxy-5-(5-methyl-1H-pyrrol-2-yl-methylene)-1,5-dihydro-pyrrol-2-on-
e (7.7 g; 27.49 mmols) as a 4:1 Z:E mixture. Yield: 75%
[0136] .sup.1NMR (400 mhz, CDCl.sub.3), ppm: 1.73 (3H, s, E), 2.19
(3H, s, Z), 5.07 (2H, s, Z), 5.16 (2H, s, E), 5.22 (1H, s, Z), 5.46
(1H, s, E), 5.67 (1H, t, J=3.0 Hz, E), 5.80 (1H, t, J=3.0 Hz, Z),
6.08 (1H, s, Z), 6.09 (1H, s, E), 6.20 (1H, t, J=3.0 Hz, E), 6.46
(1H, t, J=3.0 Hz, Z), 7.2-7.6 (5H, m, E+Z), 9.34 (1H, s, E+Z), 9.86
(1H, bs, E), 10.69 (1H, bs, Z).
EXAMPLE 2
Compound (II)
[0137] To a suspension of
4-benzyloxy-5-(5-methyl-1H-pyrrol-2-yl-methylene-
)-1,5-dihydro-pyrrol-2-one (3 g; 10.70 mmols) in dichloromethane
(160 ml) at -10.degree. C. trifluoromethansulphonic anhydride (2.16
ml; 12.84 mmols) was added dropwise under nitrogen atmosphere.
After stirring at this temperature for 30' the reaction mixture was
poured into a cold 5% NaHCO.sub.3 solution (300 ml) and extracted
with dichloromethane (2.times.100 ml). The collected organic
extracts were shaken with brine, anhydrified over anhydrous sodium
sulphate and evaporated to dryness to give
2-trifluoromethane-sulphonyloxy-4-benzyloxy-5-[(5-methyl-2H-pyrrol-2-
-ylidene)methyl]-1H-pyrrole (4.23 g; 10.26 mmols) as a yellow
solid.
[0138] Yield: 96%.
[0139] .sup.1NMR (400 mhz, CDCl.sub.3), ppm: 2.67 (3H, s), 5.05
(2H, s), 5.46 (1H, s), 6.04 (1H, d, J=3.7 Hz), 6.64 (1H, d, J=3.7
Hz), 7.08 (1H, s), 7.41 (5H, m), 10.8 (1H, bs).
EXAMPLE 3
Compound (I)
[0140] An oxygen free solution of
2-trifluoromethane-sulphonyloxy-4-benzyl-
oxy-5-[(5-methyl-2H-pyrrol-2-ylidene)methyl]-1H-pyrrole (6.0 g;
14.70 mmols) in dioxane (350 ml) was treated in sequence, under
argon atmosphere, with (1-t-butoxycarbonyl-pyrrol-2-yl)boronic acid
(12.4 g; 58.78 mmols), potassium carbonate (16.24 g; 117.57 mmols),
tetrakis (triphenylphosphine)palladium(0) (849 mg; 0.734 mmols) and
heated to 95.degree. C., under stirring, for 5 hours. After
cooling, the reaction mixture was poured into ice-water (400 ml)
and extracted with ethyl acetate (3.times.300 ml). The organic
phase was shaken with water and brine, anhydrified over anhydrous
sodium sulphate, filtered and evaporated to dryness in vacuum. The
residue was purified over a short Al.sub.2O.sub.3 column (activity
II-III) using hexane/ethyl acetate 3/1 as eluant. The collected
fractions were concentrated, dissolved in ethyl ether, treated with
a solution of hydrochloric acid in isopropyl ether at 5.degree. C.
and, after stirring for 30 minutes, filtered to give
4-benzyloxy-5-[(5-methyl-2H-pyrrol-2-ylidene)methyl]-2,2'-bi-1H-pyrrole,
hydrochloride (2.54 g; 6.94 mmols).
[0141] Yield: 47%.
[0142] .sup.1NMR (400 mhz, CDCl.sub.3), ppm: 2.56 (3H, s), 5.22
(2H, s), 6.15 (1H, d, J=1.7 Hz), 6.18 (1H, dd, J=4.0, 1.8 Hz), 6.37
(1H, m), 6.83 (1H, dd, J=4.0, 2.4 Hz), 6.94 (1H, m), 7.06 (1H, s),
7.26 (1H, m), 7.45 (5H, m), 12.65 (1H, bs), 12.81 (1H, bs), 12.85
(1H, bs)
[0143] By analogous procedure the following compounds can be
synthesized:
[0144]
4-benzyloxy-5-[(5-pentyl-2H-pyrrol-2-ylidene)methyl]-2,2'-bi-1H-pyr-
role, hydrochloride
[0145] .sup.1NMR (400 mhz, CDCl.sub.3), ppm: 0.90 (3H, m), 1.38
(4H, m), 1.78 (2H, m),2.95 (2H, t), 5.22 2H, s), 6.15 (1H, d, J=1.8
Hz), 6.20 (1H, dd, J=4.0, 1.8 Hz), 6.36 (1H, m), 6.84 (1H, dd,
J=4.0, 3.0 Hz), 6.94 (1H, m), 7.06 (1H, s), 7.24 (1H, m), 7.44 (5H,
m), 12.68 (1H, bs), 12.75 (1H, bs), 12.93 (1H, bs)
[0146] 4-benzyloxy-5-[(5-ethyl-2H-pyrrol-2-ylidene)
methyl]-2,2'-bi-1H-pyrrole, hydrochloride
[0147] .sup.1H-NMR (400 Mhz, CDCl.sub.3), ppm: 1.36 (3H, t, J=7.6
Hz), 2.99 (2H, q, J=7.6 Hz), 5.22 (2H, s,), 6.14 (1H, d, J=1.5 Hz),
6.22 (1H, dd, J=3.9, 1.4 Hz), 6.36 (1H, m), 6.85 (1H, dd, J=3.9,
2.7 Hz), 6.94 (1H, m), 7.06 (1H, s), 7.26 (1H, m), 7.45 (H, m),
12.6 (1H, bs), 12.7 (1H, bs), 12.8 (1H, bs);
[0148]
4-benzyloxy-5-[(5-propyl-2H-pyrrol-2-ylidene)methyl]-2,2'-bi-1H-pyr-
role, hydrochloride;
[0149]
4-benzyloxy-5-[(5-isopropyl-2H-pyrrol-2-ylidene)methyl]-2,2'-bi-1H--
pyrrole, hydrochloride;
[0150]
4-benzyloxy-5-[(5-butyl-2H-pyrrol-2-ylidene)methyl]-2,2'-bi-1H-pyrr-
ole, hydrochloride;
[0151]
5'-methyl-4-benzyloxy-5-[(5-methyl-2H-pyrrol-2-ylidene)methyl]-2,2'-
-bi-1H-pyrrole, hydrochloride;
[0152]
5'-methyl-4-benzyloxy-5-[(5-ethyl-2H-pyrrol-2-ylidene)methyl]-2,2'--
bi-1H-pyrrole, hydrochloride;
EXAMPLE 4
[0153] Formulation: capsules (150 mg).
[0154] Capsules, each weighing 400 mg and containing 150 mg of the
active substance, are manufactured as follows.
[0155] Composition:
3 bi-1H-pyrrole, hydrochloride 150 mg Lactose 198 mg Corn starch 50
mg Magnesium stearate 2 mg Total 400 mg
[0156] Encapsulated in two-piece hard gelatin capsules.
* * * * *