U.S. patent application number 12/007694 was filed with the patent office on 2008-11-20 for pyrazolidinol compounds.
This patent application is currently assigned to A-Viral AS. Invention is credited to Jo Klaveness, Enok Tjotta.
Application Number | 20080287390 12/007694 |
Document ID | / |
Family ID | 10856272 |
Filed Date | 2008-11-20 |
United States Patent
Application |
20080287390 |
Kind Code |
A1 |
Tjotta; Enok ; et
al. |
November 20, 2008 |
Pyrazolidinol compounds
Abstract
The invention provides the use of an optionally
hydroxyl-protected 4-hydroxy or hydroperoxy-3,5-dioxopyrazolidine
or an equivalent wherein a pyrazolidine ring attached oxygen is
replaced by a sulphur, or a physiologically acceptable salt
thereof, for the manufacture of a medicament for use in drug
therapy or prophylaxis. Additionally, the invention provides a
method of combating HIV infection which comprises administering to
an HIV-infected patient a T-lymphocyte growth suppressing agent,
preferably a pyrazolidinol, in an amount sufficient to suppress
T-lymphocyte growth in said patient for a period sufficient to
reduce the T-lymphocyte concentration in lymph nodes in said
patient.
Inventors: |
Tjotta; Enok; (Oslo, NO)
; Klaveness; Jo; (Oslo, NO) |
Correspondence
Address: |
BACON & THOMAS, PLLC
625 SLATERS LANE, FOURTH FLOOR
ALEXANDRIA
VA
22314-1176
US
|
Assignee: |
A-Viral AS
Oslo
NO
|
Family ID: |
10856272 |
Appl. No.: |
12/007694 |
Filed: |
January 14, 2008 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11371226 |
Mar 9, 2006 |
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12007694 |
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10701498 |
Nov 6, 2003 |
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11371226 |
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10019229 |
May 29, 2002 |
6852749 |
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PCT/GB00/02513 |
Jun 29, 2000 |
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10701498 |
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Current U.S.
Class: |
514/46 ; 514/220;
514/253.01; 514/404; 514/49 |
Current CPC
Class: |
A61K 31/495 20130101;
C07D 231/30 20130101; A61K 31/7076 20130101; A61P 5/38 20180101;
A61P 15/08 20180101; C07D 231/32 20130101; A61P 3/10 20180101; A61P
21/04 20180101; A61P 31/08 20180101; A61K 31/551 20130101; A61K
31/55 20130101; A61P 29/00 20180101; A61K 31/415 20130101; A61K
31/415 20130101; A61K 2300/00 20130101; A61K 2300/00 20130101; A61P
7/00 20180101; A61K 31/55 20130101; A61K 31/4152 20130101; A61P
25/28 20180101; A61P 1/04 20180101; A61K 31/495 20130101; A61K
31/495 20130101; A61P 21/00 20180101; A61P 31/18 20180101; A61K
31/55 20130101; A61P 31/12 20180101; A61K 31/415 20130101; A61P
7/06 20180101; A61K 45/06 20130101; A61K 31/522 20130101; A61P
37/02 20180101; A61K 31/70 20130101; A61P 37/06 20180101 |
Class at
Publication: |
514/46 ; 514/404;
514/49; 514/253.01; 514/220 |
International
Class: |
A61K 31/7064 20060101
A61K031/7064; A61K 31/4152 20060101 A61K031/4152; A61K 31/496
20060101 A61K031/496; A61P 31/12 20060101 A61P031/12; A61K 31/551
20060101 A61K031/551; A61K 31/7076 20060101 A61K031/7076 |
Foreign Application Data
Date |
Code |
Application Number |
Jun 29, 1999 |
GB |
9915184.7 |
Claims
1. (canceled)
2. A method of treatment of the human or non-human body to combat
an inflammatory autoimmune or viral disease or a tumor, which
method comprises administering to said body an optionally
hydroxy-protected 4-hydroxy or hydroperoxy-3,5-dioxo-pyrazolidine
or an equivalent wherein a pyrazolidine ring attached oxygen is
replaced by a sulphur, or a physiologically acceptable salt
thereof.
3. A method as claimed in claim 2 comprising administering said
optionally hydroxy-protected 4-hydroxy or
hydroperoxy-3,5-dioxo-pyrazolidine or an equivalent wherein a
pyrazolidine ring attached oxygen is replaced by a sulphur, or a
physiologically acceptable salt thereof in combination with another
agent.
4. A method as claimed in claim 27 wherein said additional
antiviral agent is at least one antiviral agent selected from a
reverse transcriptase inhibitor and a protease inhibitor.
5. A method as claimed in claim 3 wherein said additional antiviral
agent is an agent selected from the group of AZT, indinavir,
nevirapine and 2',3'-dideoxyinosine (daI).
6. (canceled)
7. A method of combating HIV infection which comprises
administering to an HIV-infected patient a T-lymphocyte growth
suppressing agent in an amount sufficient to suppress T-lymphocyte
growth in said patient for a period sufficient to reduce the
T-lymphocyte concentration in the lymphatic system in said patient
by at least 25% said administration being repeated at intervals of
at least 3 months.
11-26. (canceled)
27. A method as claimed in claim 2 wherein said disease is a
disease caused by a pathogen from the group of togaviridea,
reoviridea, picornaviridea, hantaviridea, orthomyxoviridea,
paramyxoviridea, mononegaviralis, viral hepatitis, haemorrhagic
fevers, flaviviridea, viral encephalitis, coronaviridea,
calciviridea, adenoviridea, papovaviridea, arboviridea, pox virus,
rhabdoviridea, arenaviridea HIV-1, HIV-2, HTLV-I, HTLV-II and
herpes viruses.
28. A method of combating HIV infection as claimed in claim 7
wherein said T-lymphocyte growth suppressing agent is a
pyrazolidinol.
29. A method as claimed in claim 7 wherein said interval is at
least 9 months.
30. A method as claimed in claim 7 wherein a 4-hydroxy or
hydroperoxy-3,5-dioxo-pyrazolidine or an equivalent wherein a
pyrazolidine ring attached oxygen is replaced by a sulphur, or a
physiologically acceptable salt thereof is administered in a daily
dose of 0.1 to 10 .mu.mol/kg bodyweight.
31. A method as claimed in claim 7 wherein a 4-hydroxy or
hydroperoxy-3,5-dioxo-pyrazolidine or an equivalent wherein a
pyrazolidine ring attached oxygen is replaced by a sulphur, or a
physiologically acceptable salt thereof is administered in a daily
dose of 0.1 to 10 pmol/kg bodyweight.
32. A pharmaceutical composition comprising an optionally
hydroxy-protected 4-hydroxy or hydroperoxy3,5-dioxo-pyrazolidine or
an equivalent wherein a pyrazolidine ring attached oxygen is
replaced by a sulphur, or a physiologically acceptable salt
thereof, together with at least one pharmaceutically acceptable
carrier or excipient.
33. A method of treatment of the human or non-human body to combat
an autoimmune disease or tissue rejection, which method comprises
administering to said body an optionally hydroxy-protected
4-hydroxy or hydroperoxy-3,5-dioxo-pyrazolidine or an equivalent
wherein a pyrazolidine ring attached oxygen is replaced by a
sulphur, or a physiologically acceptable salt thereof.
34. A method of claim 33 wherein said disease is selected from
Addison's disease, Behcet's syndrome, diabetes mellitus, haemolytic
anaemia, lupus erythematosus, multiple sclerosis, myasthenia
gravis, pernicious anaemia, polyglandular deficiency, polymyositis,
dermatomyositis, testicular failure, thrombocytopenic purpura,
Crohns disease, ulcerative colitis and rheumatoid arthritis.
35. A method of claim 24 wherein said tissue rejection is tissue
rejection following transplant.
36. The method of claim 3 to combat a viral disease wherein said
another agent is an antiviral agent.
37. The method of claim 2 wherein said tumour is a T-cell tumour or
Karposi's sarcoma.
38. The method of claim 27 wherein said T-cell tumour is selected
from Sezary Syndrome, mycosis fungoides, T-cell lymphoma, and CD4
cell tumours.
Description
[0001] The present invention relates to certain pyrazolidinols and
their sulphur (i.e. thio/thiol) analogs and pharmaceutical
compositions thereof for use in antiviral, e.g. anti-HIV therapy
and as anti-inflammatories and immunomodulators.
[0002] Phenbutazone and oxyphenbutazone are 1,2-bis
aromatic-3,5-pyrazolidinediones which have been used as
non-steroidal anti-inflammatory drugs (NSAIDs)
##STR00001##
[0003] Other 3,5-pyrazolidinediones have likewise been proposed for
use as NSAIDs (see for example U.S. Pat. No. 3,968,219 (Rahtz)) and
the hydroxy-protected enol forms have been proposed as pro-drug
forms of phenbutazone and oxyphenbutazone (see U.S. Pat. No.
4,117,232 (Bodor), U.S. Pat. No. 3,957,803 (Bodor), U.S. Pat. No.
4,169,147 (Bodor), U.S. Pat. No. 4,036,845 (Bodor) and U.S. Pat.
No. 4,139,709 (Bodor)).
[0004] In U.S. Pat. No. 4,956,377 (Miesch) it was proposed that
this class of NSAIDs had utility as an antiviral agent, in
particular for the treatment of HIV.
[0005] We have now surprisingly found that where the 4-carbon of
the N.sub.2C.sub.3 ring carries an optionally protected hydroxy or
thiol group, the compounds have very significantly enhanced
antiviral, in particular anti-HIV, efficacy.
[0006] Thus viewed from one aspect the invention provides the use
of an optionally hydroxy-protected 4-hydroxy or
hydroperoxy-3,5-dioxo-pyrazolidine or an equivalent wherein a
pyrazolidine ring attached oxygen is replaced by a sulphur, or a
physiologically acceptable salt thereof, for the manufacture of a
medicament for use in therapy or prophylaxis.
[0007] Where a particular 4-hydroxy or
hydroperoxy-3,5-dioxo-pyrazolidine may exist in more than one
stereoisomeric form, it may be used in single isomer form or as an
isomer mixture, e.g. a racemic mixture.
[0008] Viewed from a further aspect, the invention provides an
optionally hydroxy-protected 4-hydroxy or
hydroperoxy-3,5-dioxo-pyrazolidine or an equivalent wherein a
pyrazolidine ring attached oxygen is replaced by a sulphur, or a
physiologically acceptable salt thereof.
[0009] Viewed from a still further aspect the invention provides a
method of treatment of the human or non-human (e.g. mammalian,
reptilian or avian) body to combat an inflammatory or viral
disease, preferably an immuno-deficiency viral disease, in
particular HIV, which method comprises administering to said body
an optionally hydroxy-protected 4-hydroxy or
hydroperoxy-3,5-dioxo-pyrazolidine or an equivalent wherein a
pyrazolidine ring attached oxygen is replaced by a sulphur, or a
physiologically acceptable salt thereof.
[0010] Viewed from a still further aspect, the invention provides a
pharmaceutical composition comprising an optionally
hydroxy-protected 4-hydroxy or hydroperoxy-3,5-dioxo-pyrazolidine
or an equivalent wherein a pyrazolidine ring attached oxygen is
replaced by a sulphur, or a physiologically acceptable salt
thereof, together with at least one pharmaceutically acceptable
carrier or excipient.
[0011] The applicants have found that oxyphenbutazone, as
commercially available, contains minute quantities of certain
impurities, presumably as a result of undesired oxidative
breakdown. One of these, present at about 0.4% wt, is
4-butyl-4-hydroxy-2(p-hydroxyphenyl)-1-phenyl-3,5-pyrazolidinedione
(hereinafter "4-OH--OPB"), i.e.
##STR00002##
4-OH--OPB is of course a compound according to the invention and
thus it should be understood that references to the 4-hydroxy
compounds of the invention, their use and compositions thereof
should not be taken to include references to such compounds when in
intimate admixture with overwhelmingly larger quantities of a
3,5-pyrazolidinedione which carries no optionally protected
4-hydroxy or 4-thiol group. By overwhelmingly larger is meant a
relative weight ratio of at least 98:2. In general, the compounds
of the invention should not desirably be used in intimate admixture
with larger quantities (i.e. a relative weight ratio of more than
50:50) of such compounds carrying no O or S attached group at the
4-position, and more desirably they should not be used with such
compounds present in greater than 10:90 weight ratio.
[0012] The compounds of the invention, hereinafter referred to as
pyrazolidinols for convenience, will preferably be of formula I
##STR00003##
(where each X.sub.2, which may be the same or different is O or S,
preferably O; X.sub.1 is O, OO or S, preferably O or S, most
preferably O; R.sub.1 is hydrogen or a hydroxyl or thiol protecting
group (e.g. an acyl group, preferably containing up to 6 carbons,
e.g. an acyl group such as an alkylcarbonyl group, for example
acetyl), preferably hydrogen; R.sub.2 is hydrogen or more
preferably a carbon attached organic group containing up to 10
carbons, e.g. an alkyl, alkenyl, alkynyl, alkaryl, aralkyl or
aralkenyl group, optionally substituted, e.g. by a sulphonyl group;
and each Ar, which may be the same or different, is a homo or
heterocyclic aromatic group, optionally substituted, e.g. by
C.sub.1-6 alkyl or alkoxy groups) or a salt thereof.
[0013] In the compounds of the invention 0, 1 or 2 of the X.sub.1
and X.sub.2 groups may be S. It is thought that it is especially
preferred that one thio X.sub.2 group be present.
[0014] In the compounds of the invention, the R.sub.2 group is
preferably other than hydrogen and may for example be straight
chain, branched, cyclic or cyclic-attached-to-straight chain.
Preferably it is an alkyl or alkenyl group, especially a C.sub.1-6
alkyl or alkenyl group, e.g. n-propyl, n-butyl, n-pentyl or
1-methyl-but-2-en-4-yl or an aralkyl (e.g. benzyl) or alkaryl (e.g.
methylphenyl) or arylsulphonylalkyl (eg phenylsulphonylethyl)
group.
[0015] Where R.sub.1 in the compounds of the invention is other
than hydrogen it is preferably a metabolically labile hydroxy- or
thiol-protecting group which yields a physiologically tolerable
R.sub.1OH metabolite. Acyl groups are preferred in this regard.
[0016] In the compounds of formula I, where each X.sub.2 is oxygen
and one Ar is phenyl, the other Ar is preferably other than phenyl
e.g. parahydroxyphenyl.
[0017] A wide range of hydroxy- and thiol-protecting groups however
is known from the literature (see McOmie, "Protective groups in
organic chemistry", Plenum, 1973 and Greene, "Protective groups in
organic synthesis", Wiley Interscience, NY, 1981) and many
compounds of formula I in which R.sub.1 is a protecting group may
be useful as intermediates in the production of compounds of
formula I in which R.sub.1 is hydrogen.
[0018] The Ar groups in the compounds of formula I are preferably 5
to 7 membered aromatic rings, optionally carrying a fused aromatic
ring and optionally substituted on ring atoms, for example by
C.sub.1-6 alkyl groups but especially by electron withdrawing
substituents, e.g. hydroxy, thiol, phenyl, C.sub.1-6 alkoxy, cyano,
halo (e.g. Cl, F, Br or I), protected hydroxy, or protected thiol.
Ring heteroatoms will generally be selected from O, N and S,
preferably with a single ring heteroatom in any aromatic Ar
heterocycle. Ar is preferably phenyl optionally substituted,
especially in the para-position by --X.sub.1--R.sub.1, or Cl (where
--X.sub.1--R.sub.1 is as defined above). Especially preferably one
Ar is phenyl and the other is p-hydroxy-phenyl.
[0019] Where the substitution of the pyrazolidinols of the
invention is such that they may form addition salts with acids or
bases, the addition salts which have physiologically tolerable
counterions are of course preferred, e.g. sodium, organic amine,
halides, phosphates, hydrogen carbonates, etc.
[0020] The pyrazolidinols of the invention may particularly
advantageously be used in combination therapy with other antiviral,
especially anti-HIV, agents, in particular reverse transcriptase
inhibitors and/or protease inhibitors, e.g. zidovudine, didanovine,
zalcitabine, stavudine, lamivudine, nevirapine, delavirdine,
indinavir, ritonavir, nelfinavir, hydroxyurea kolchicine, AZT and
2',3'-dideoxyinosine (ddI). Such combination therapy forms a
further aspect of the present invention.
[0021] A drawback of traditional combination therapy, has often
been that even under intensive antiviral treatment with a
combination of drugs, a little HIV production continues and is
unaffected by treatment. The compounds of the invention may prove
to have an effect in reducing this residual HIV production when
given in combination with other antiviral agents. This may be due
to the increasing antiviral effect which has been seen in long term
cell culture experiments and which may counteract any development
of resistance to the compounds.
[0022] The pyrazolidinols of the invention may be prepared by
oxidation of a corresponding compound where R.sub.1X.sub.1 is
replaced by hydrogen; by reaction of a corresponding compound where
R.sub.1X.sub.1 is HX.sub.1 with a hydroxy or thiol protecting agent
to introduce a non-hydrogen R.sub.1 group; or by condensation of a
hydrazine derivative with an optionally protected 2-hydroxy-propane
dioic acid ester (or a sulphur analog), e.g. by condensation of a
compound of formula II
Ar--HN--NH--Ar (II)
with a compound of formula III
##STR00004##
where R.sub.1, R.sub.2, X.sub.1, X.sub.2 and Ar are as hereinbefore
defined and X.sub.2R.sub.3 is a leaving group, for example where
R.sub.3 is an alkyl group, e.g. a C.sub.1-6 alkyl group.
[0023] Alternatively, a compound of formula. II may be condensed
with a compound of formula IV
##STR00005##
(where X.sub.2 and R.sub.3 are as defined above) and then reacted
with an alkylating agent, e.g. (R.sub.2).sub.2Zn to produce a
compound of formula I in which X.sub.1R.sub.1 is OH or SH.
[0024] For administration, the pyrazolidinols of the invention may
be formulated in any convenient form, e.g. tablets, coated tablets
(e.g. delayed release tablets), capsules, solutions, suspensions,
dispersions, syrups, powders, sprays, suppositories, transdermal
patches, gels, emulsions and creams. Administration may be via any
convenient route, e.g. oral, rectal, transdermal, nasal,
subcutaneous, intravenous, intramuscular, etc. Oral administration,
e.g. of tablets or capsules is preferred. The pyrazolidinols may be
formulated together with conventional pharmaceutical carriers,
diluents or excipients, e.g. aqueous carriers (for example water
for injections), binders, fillers, stabilizers, osmolality
adjusting agents, effervescing agents, pH modifiers, viscosity
modifiers, sweeteners, lubricants, emulsifiers, flavours, coating
agents (e.g. gastric juice resistant coatings), etc. Where any
formulation results in a loss of compound, this loss should be
calculated and the dosage increased proportionally to obtain the
desired active concentration.
[0025] The dosage of the pyrazolidinols given according to the
invention will depend on the size and species of the subject being
treated but will generally be in the range of 0.05 to 2000 mg/day,
more particularly 0.5 to 1000 mg/day, especially 1 to 100 mg/day,
preferably with administration being effected once, twice, three
times or four times daily. For mice, doses of up to 2000 mg/kg
(corresponding to 20 mM maximal concentration in extracellular
fluid) could be given before lethal dosage was reached, ie
effective treatment doses were up to 200000 times smaller than the
lethal dose.
[0026] For regular, e.g. continuous daily treatment according to
the invention, the daily dosage of the pyrazolidinol will
preferably be in the range 5 nmol to 2 .mu.mol/kg bodyweight, more
preferably 100 nmol to 1.5 .mu.mol/kg, especially 500 nmol to 1
.mu.mol/kg.
[0027] Inhibition of virus production may be achieved by small
intermittent doses of pyrazolidinol, and are expected to induce
inhibition of the virus after a latency of about 11 weeks.
Subsequently, inhibition may be expected to level out, should
resistance to the compound develop. Such doses may be administered
at a frequency of 1-14 days, preferably 7 days. The doses should be
equivalent to a concentration in plasma/tissue fluid of from
100-1000 nM and may be obtained by ingestion or injection of from
0.7-7 mg in a 70 Kg human.
[0028] However, in a particularly preferred embodiment of the
invention, a pyrazolidinol according to the invention is
administered at a dose sufficient to suppress T-lymphocyte (CD4 and
CD8 cell) growth (e.g. a daily dose of 0.1 to 10 .mu.mol/kg) for a
period of 1 to 14 days, preferably 2 to 7 days at intervals of at
least 3 months, preferably at least 9 months, e.g. 10 to 18 months.
In this way the patient's immune system may be "refreshed" by
removal of the preponderance of T-lymphocytes directed to HIV
antigens. Such a treatment indeed is novel and forms a further
aspect of the invention. Viewed from this aspect the invention
provides a method of combating HIV infection which comprises
administering to an HIV-infected patient a T-lymphocyte growth
suppressing agent, e.g. a pyrazolidinol, in an amount sufficient to
suppress T-lymphocyte growth in said patient for a period
sufficient to reduce the T-lymphocyte concentration in the
lymphatic system, e.g. the lymph nodes, in said patient by at least
25%, more preferably at least 50%, said administration being
repeated at intervals of at least 3 months, preferably at least 9
months.
[0029] High tissue concentrations intended to give an
immunomodulating effect should preferably be given for limited
periods at doses of 1 .mu.M or above in plasma/tissue fluid. Such
doses and lengths of administration will vary according to the
condition of each patient and may be decided with the guidance of
tests such as the count of HIV memory subsets of T8 and T4. As
stated above, the goal of treatment according to this aspect of the
invention should be to reduce subsets which are found to be too
prevalent without overly affecting naive T-cells.
[0030] In order to obtain the desired reduction of HIV specific
lymphocytes (e.g. HIV memory CD8 and CD4 lymphocytes) without
overly affecting naive T-lymphocytes or other essential blood
cells, monoclonal antibodies against the unwanted subtypes may also
be administered. Further, drugs such as kolchicine and/or
hydroxy-urea may be included in the intermittent intensive
treatment. Such additional drugs are anticipated to have a somewhat
different immunomodulating effect to the compounds of the invention
and so may be used advantageously in combination with
pyrazolidinols for refreshing the immune system.
[0031] Besides HIV, the pyrazolidinols of the invention may be used
to combat other viral infections, especially retroviral infections
but also infections by togaviridea, reoviridea, picornaviridea,
hantaviridea, orthomyxoviridea, paramyxoviridea, mononegaviralis,
viral hepatitis, haemorrhagic fevers, flaviviridea, viral
encephalitis, coronaviridea, calciviridea, adenoviridea,
papovaviridea, arboviridea, pox virus, rhabdoviridea, herpes virus
and arenaviridea. The pyrazolidinols of the invention may in
particular be used to combat viral infections of CD4 cells, e.g.
HIV-1, HIV-2, HTLV-I, HTLV-II and herpes viruses, for example to
combat AIDS, T-cell tumours (e.g. Sezary Syndrome, mycosis
fungoides and T-cell lymphoma, and particularly CD4 cell tumours),
tropic spastic paraparesis, and Karposi's sarcoma. Moreover despite
not being of the accepted formula for NSAIDs (which would require
an acid proton in place of R.sub.1X.sub.1 at the 4-position), they
may be used as anti-inflammatory drugs. All these uses form aspects
of the invention.
[0032] Due to the immunomodulating effect of the compounds of the
invention, they are expected to have uses in control of other
immune-system related diseases, such as auto immune diseases and as
immunosuppressants. In particular, the compounds of the invention
are expected to have a positive effect on the generation of
autoimmune diseases, on developed autoimmune diseases and on
diseases related to such diseases, such as Addison's disease,
Behcet's syndrome, diabetes mellitus and other endocrine diseases,
haemolytic anaemia, lupus erythematosus, multiple sclerosis,
myasthenia gravis, pernicious anaemia, polyglandular deficiency,
polymyositis, dermatomyositis, testicular failure, thrombocytopenic
purpura, Crohns disease, ulcerative colitis, rheumatic disorders
(e.g. rheumatoid arthritis) etc.
[0033] The effect of the compounds of the invention on the immune
system may also be that of immunosuppression. Such an effect may be
used, for example, to control rejection of a medical transplant or
implant. In particular, the compounds may be used to reduce
rejection following tissue or organ transplant.
[0034] Various 4-hydroxy-3,5-dioxo-pyrazolidines are known in the
literature (although not for medical purposes such as HIV therapy).
These are compounds of formula V
##STR00006##
where R.sub.a to R.sub.d are as set out in Table 1 below:
TABLE-US-00001 TABLE 1 R.sub.a R.sub.b R.sub.c R.sub.d H H H
C.sub.6H.sub.5 H H C.sub.6H.sub.5 C.sub.6H.sub.5 CH.sub.3 H H
C.sub.6H.sub.5 CH.sub.3 H H --CH.sub.2--C.sub.6H.sub.5 CH.sub.3 H H
p-CH.sub.3O--C.sub.6H.sub.4 CH.sub.3 H H p-Cl--C.sub.6H.sub.4
C.sub.2H.sub.5 H H C.sub.6H.sub.5 C.sub.2H.sub.5 H C.sub.6H.sub.5
C.sub.6H.sub.5 C.sub.2H.sub.5 H H N-methyl-piperidin-4-yl
iC.sub.3H.sub.7 H H C.sub.6H.sub.5 nC.sub.3H.sub.7 H H
C.sub.6H.sub.5 nC.sub.3H.sub.7 H C.sub.6H.sub.5 C.sub.6H.sub.5
nC.sub.3H.sub.7 H H 5-phenyl-triazol-1-yl C.sub.4H.sub.9 H H
C.sub.6H.sub.5 C.sub.4H.sub.9 H C.sub.6H.sub.5 C.sub.6H.sub.5
C.sub.4H.sub.9 H C.sub.6H.sub.5 p-OH--C.sub.6H.sub.4 C.sub.4H.sub.9
OH C.sub.6H.sub.5 C.sub.6H.sub.5 C.sub.4H.sub.9 OH C.sub.6H.sub.5
p-OH--C.sub.6H.sub.4 C.sub.4H.sub.9 H H N-methyl-piperidin-4-yl
C.sub.5H.sub.11 H H C.sub.6H.sub.5 C.sub.5H.sub.11 H C.sub.6H.sub.5
C.sub.6H.sub.5 C.sub.5H.sub.11 H H 5-phenyl-triazol-1-yl Cyclohexyl
H H C.sub.6H.sub.5 Phenyl H H C.sub.6H.sub.5 Phenyl H
C.sub.6H.sub.5 C.sub.6H.sub.5 Benzyl H H C.sub.6H.sub.5 Benzyl H
C.sub.6H.sub.5 C.sub.6H.sub.5 CH.sub.3CO(CH.sub.2).sub.2 H
C.sub.6H.sub.5 C.sub.6H.sub.5 (CH.sub.3).sub.2C.dbd.CH-- H
C.sub.6H.sub.5 C.sub.6H.sub.5 (CH.sub.2).sub.2C.dbd.CHCH.sub.2 H
C.sub.6H.sub.5 C.sub.6H.sub.5 C.sub.6H.sub.5SCH.sub.2CH.sub.2 H
C.sub.6H.sub.5 C.sub.6H.sub.5 Pyrrolidin-1-yl H C.sub.6H.sub.5
C.sub.6H.sub.5 Piperidin-1-yl H C.sub.6H.sub.5 C.sub.6H.sub.5
Morpholin-4-yl H C.sub.6H.sub.5 C.sub.6H.sub.5
Such compounds are thus not claimed per se herein; however their
use and pharmaceutical compositions containing them do form part of
the scope of the invention.
[0035] The invention will now be illustrated further by the
following non-limiting Examples and by reference to the Figures, in
which:
[0036] FIG. 1 shows the HIV antigen concentration in human CD4
cells infected with HIV and treated with 4-butyl-4-hydroxy-2
(p-hydroxyphenyl)-1-phenyl-3,5-pyrazolidinedione (4OH--OPB) or
phenbutazone (PB) at various concentrations;
[0037] FIG. 2 shows the effect of
4-butyl-4-hydroxy-2(p-hydroxyphenyl)-1-phenyl-3,5-pyrazolidinedione
(4OH--OPB) when used in combination with AZT;
[0038] FIG. 3 shows the effect of
4-butyl-4-hydroxy-2(p-hydroxyphenyl)-1-phenyl-3,5-pyrazolidinedione
(4OH--OPB) when used in combination with indinavir; and;
[0039] FIG. 4 shows the effect of
4-butyl-4-hydroxy-2(p-hydroxyphenyl)-1-phenyl-3,5-pyrazolidinedione
(4OH--OPB) when used in combination with nevirapine;
[0040] A similar effect to those shown in FIGS. 2-4 is seen when
OPB is used in combination with 2',3'-dideoxyinosine (daI).
EXAMPLE 1
Preparation of 4-Methoxyazobenzene
[0041] A mixture of 4-phenylazophenol (9.9 g; 50 mmol), iodomethane
(7.1 g; 50 mmol), potassium carbonate (6.9 g; 50 mmol), and acetone
(100 ml) was refluxed 48 h. After evaporating off the solvent, the
residue was dissolved in water (25 ml), diethyl ether (50 ml) and
THF (30 ml). The aqueous layer was extracted with ether (3.times.20
ml) and the combined organic solutions were washed with saturated
NaCl solution (1.times.20 ml) and dried (MgSO.sub.4). After
filtration and evaporation, the residue was recrystallized from 96%
ethanol to give 8.7 g (82%).
EXAMPLE 2
Preparation of 1-(4-Methoxyphenyl)-2-phenylhydrazine
[0042] Zinc powder (10.0 g; 0.15 mol) was added to a stirred
mixture of 4-methoxyazobenzene (4.24 g; 20.0 mmol) in 96% ethanol
(75 ml) and saturated NH.sub.4Cl solution (2.0 ml) at 0.degree. C.
(bath temperature). Two more portions of saturated NH.sub.4Cl
solution (2.0 ml) were added at 1.5 h intervals. The yellowish
solution was poured into cold water (100 ml) and filtered. The
residue was extracted with methylene chloride (5.times.50 ml). The
combined aqueous phases were extracted with methylene chloride
(3.times.25 ml). The combined organic solutions were dried
(Na.sub.2SO.sub.4), filtered, and evaporated to give 4.3 g crude
1-(4-methoxyphenyl)-2-phenylhydrazine as a reddish oil.
EXAMPLE 3
Preparation of
4-(1-Butyl)-1-(4-methoxyphenyl)-2-phenyl-3,5-pyrazolidinedione
[0043] Diethyl butylmalonate (4.33 g; 20.0 mmol) was added to a
stirred solution of sodium (0.46 g; 20.0 mmol) in absolute ethanol
(20 ml), followed by crude 1-(4-methoxyphenyl)-2-phenylhydrazine
(4.3 g; 20 mmol max.) in absolute ethanol (5 ml). About 2/3 of the
ethanol was distilled off and xylene (20 ml) was added to the
residue. The reaction mixture was heated to 140-145.degree. C.
(bath temperature) for 15 h to distill off the rest of the ethanol.
The reaction mixture was cooled to 0.degree. C. (bath temperature)
and poured into ice water (ca. 100 ml). The aqueous layer was
extracted with CH.sub.2Cl.sub.2 (2.times.15 ml); the extracts were
discarded. The cold aqueous layer was acidified with 6 M HCl (5 ml)
and extracted with CH.sub.2Cl.sub.2 (3.times.10 ml). The combined
extracts were washed with water (2.times.10 ml) and dried
(MgSO.sub.4). Filtration and evaporation gave 3.84 g amber oil.
Purified by flash chromatography on a 130.times.65 mm silica gel 60
column eluted with ethyl acetate-heptane (1:3) to give 1.45 g (21%)
colourless oil.
[0044] .sup.1H NMR (200 MHz; CDCl.sub.3): .delta. 0.90 (3H, t,
J=7.5 Hz), 1.25-1.6 (4H, m), 2.0-2.15 (2H, m), 3.37 (3H, t, J=6.0
Hz), 3.69 (3H, s), 6.81 (2H, d, J=8.4 Hz), 7.22 (2H, d, J=8.6 Hz),
7.1-7.35 (5H, m).
[0045] .sup.13C NMR (50 MHz; CDCl.sub.3): .delta. 13.6, 22.2, 27.5,
27.6, 45.6, 54.7, 112.9, 121.6, 123.3, 125.4, 127.1, 127.4, 133.9,
156.5, 168.2, 168.7.
EXAMPLE 4
Preparation of
1,2-Diphenyl-4-(4-methylphenyl)-3,5-pyrazolidinedione
[0046] Prepared from 1,2-diphenylhydrazine (3.70 g; 20.0 mmol),
diethyl 2-(p-tolyl)malonate (5.0 g; 20.0 mmol), and sodium (0.46 g;
20.0 mmol) using the procedure of Example 3. The crude product
crystallized on standing and was recrystallized twice from absolute
ethanol to give 1.22 g (18%), mp 184-185.degree. C.
[0047] .sup.1H NMR (200 MHz; CDCl.sub.3): .delta. 2.31 (3H, s),
4.51 (1H, s), 7.1-7.4 (14H, m).
[0048] .sup.13C NMR (50 MHz; CDCl.sub.3): .delta. 21.1, 51.9,
122.7, 126.9, 128.3, 129.0, 129.9, 135.8, 138.3, 168.6.
EXAMPLE 5
Preparation of 4-Benzyl-1,2-diphenyl-3,5-pyrazolidinedione
[0049] Prepared from 1,2-diphenylhydrazine (4.60 g; 25.0 mmol),
diethyl benzylmalonate (5.0 g; 20 mmol), and sodium (0.46 g; 20.0
mmol) using the procedure of Example 3. The crude product was
recrystallized from absolute ethanol to gave 3.51 g (50%), mp
136-137.degree. C. [lit. 137-138.degree. C. (Beil. III/IV, 24,
1463)].
[0050] .sup.1H NMR (200 MHz; CDCl.sub.3): .delta. 3.41 (2H, d,
J=4.6 Hz), 3.63 (1H, t, J=5.0 Hz), 6.85-7.3 (10H, m).
[0051] .sup.13C NMR (50 MHz; CDCl.sub.3): .delta. 33.9, 48.5,
123.2, 126.9, 127.3, 128.6, 128.7, 129.9, 135.2, 135.4, 169.3.
EXAMPLE 6
Preparation of 4-Allyl-1,2-diphenyl-3,5-pyrazolidinedione
[0052] Prepared from 1,2-diphenylhydrazine (5.2 g; 28.0 mmol),
diethyl allylmalonate (5.0 g; 25.0 mmol), and sodium (0.58 g; 25.0
mmol) using the procedure of Example 3. The crude product was
recrystallized from absolute ethanol to give 2.21 g (30%) tan
crystals, mp 135-137.degree. C.
[0053] .sup.1H NMR (200 MHz; CDCl.sub.3): .delta. 2.82 (2H, t,
J=6.0 Hz), 3.46 (2H, t, J=5.4 Hz), 5.1-5.3 (2H, dd), 5.7-5.95 (1H,
m), 7.1-7.3 (10H, m).
[0054] .sup.13C NMR (50 MHz; CDCl.sub.3): .delta. 31.7, 46.4,
119.9, 122.7, 126.8, 128.9, 131.7, 135.6, 169.5.
EXAMPLE 7
Preparation of
4-(1-Butyl)-4-hydroxy-1-(4-hydroxyphenyl)-2-phenyl-3,5-pyrazolidinedione
(4OH--OPB)
Method A
[0055] Oxyphenbutazone.H.sub.2O (1 mmol), 30% H.sub.2O.sub.2 (0.7
mL), 1N NaOH (0.1 mL) and methanol (3.5 mL) are allowed to stand
for 13 hours at ambient temperature. The mixture is then poured
into 5% HCl (20 mL) and extracted with ethyl acetate (2.times.20
mL). The ethyl acetate phase is separated, dried over sodium
carbonate and the solvent is removed under reduced pressure without
heating. The residue is subjected to flash chromatography
(silica/ethyl acetate). The title product is recrystallized from
ethyl acetate.
Method B
[0056] A solution of oxyphenbutazone hydrate (2.0 g; 5.8 mmol), 35%
hydrogen peroxide solution (3.4 ml; 40 mmol), and 1 M sodium
hydroxide solution (0.6 ml; 0.6 mmol) in methanol (20 ml) was
allowed to stand for 24 h at ambient temperature. The mixture was
acidified with 1 M HCl solution (50 ml) and extracted with ethyl
acetate (4.times.15 ml). The combined extracts were washed with
saturated NaCl solution (1.times.10 ml) and dried (MgSO.sub.4).
After filtration and evaporation, the residue was purified by flash
chromatography on a 100.times.65 mm silica gel 60 column eluted
with ethyl acetate-heptane (1:1), taking 50-ml fractions, giving
1.3 g (66%).
[0057] .sup.1H NMR (200 MHz; CDCl.sub.3): .delta. 0.88 (3H, t,
J=6.6 Hz), 1.25-1.5 (4H, m), 1.95-2.05 (2H, m), 6.49 (1H, br s),
6.75 (2H, d, J=8.9 Hz), 7.12 (2H, d, J=8.9 Hz), 7.1-7.35 (5H,
m).
[0058] .sup.13C NMR (50 MHz; CDCl.sub.3): .delta. 13.6, 22.3, 24.3,
36.2, 72.8, 114.3, 121.9, 123.8, 125.3, 125.7, 127.2, 133.5, 154.6,
169.0, 169.5.
EXAMPLE 8
Preparation of
4-(1-Butyl)-4-hydroxy-1-(4-methoxyphenyl)-2-phenyl-3,5-pyrazolidinedione
[0059] Prepared from
4-(1-butyl)-1-(4-methoxyphenyl)-2-phenyl-3,5-pyrazolidinedione
(1.35 g; 3.8 mmol), 35% H.sub.2O.sub.2 (4.3 ml; 50 mmol), 2 M NaOH
(0.35 ml; 0.7 mmol), and methanol (50 ml) using the procedure of
Example 7. Purified by flash chromatography on a 110.times.65 mm
silica gel 60 column eluted with ethyl acetate-heptane (1:1) to
give 0.7 g (52%).
[0060] .sup.1H NMR (200 MHz; CDCl.sub.3): .delta. 0.85 (3H, t,
J=6.2 Hz), 1.2-1.5 (4H, m), 2.0-2.1 (2H, m), 3.69 (3H, s), 4.8 (1H,
br s), 6.77 (2H, d, J=9.0 Hz), 7.19 (2H, d, J=9.0 Hz), 7.1-7.35
(5H, m).
[0061] .sup.13C NMR (50 MHz; CDCl.sub.3): .delta. 13.5, 22.3, 24.3,
36.7, 54.7, 73.3, 113.0, 122.1, 123.8, 125.8, 126.1, 127.5, 133.0,
156.7, 168.5, 169.0.
EXAMPLE 9
Preparation of
1,2-Diphenyl-4-hydroxy-4-[2-(phenylsulfonyl)ethyl]-3,5-pyrazolidinedione
[0062] Prepared from (.+-.)-sulfinpyrazone (2.02 g; 5.0 mmol), 35%
H.sub.2O.sub.2 (4.3 ml; 50 mmol), 2 M NaOH (0.35 ml; 0.7 mmol), and
methanol (50 ml) using the procedure of Example 7. Purified by
flash chromatography on a 130.times.65 mm silica gel 60 column
eluted with ethyl acetate-acetic acid (20:1) to give 80 mg
(4%).
[0063] .sup.1H NMR (200 MHz; CDCl.sub.3): .delta. 2.1-2.5 (2H, m),
3.0-3.7 (2H, m), 5.5 (1H, br s), 6.4-7.9 (15H, m).
[0064] .sup.13C NMR (50 MHz; CDCl.sub.3): .delta. 28.7, 47.5, 70.2,
121.5, 122.9, 125.8, 126.5, 127.4, 127.7, 127.9, 129.9, 133.3,
133.4, 136.9, 139.2, 167.5, 168.0.
EXAMPLE 10
1,2-Diphenyl-4-hydroxy-4-(4-methylphenyl)-3,5-pyrazolidinedione
[0065] A mixture of
1,2-diphenyl-4-(4-methylphenyl)-3,5-pyrazolidinedione (1.10 g; 3.2
mmol), 35% H.sub.2O.sub.2 (0.47 ml; 5.5 mmol), and acetic acid (40
ml) was stirred 16 days at room temperature. Sodium metabisulfite
(1.0 g) was added and excess acetic acid evaporated off. The
residue was dissolved in hot ethyl acetate (25 ml) and benzene (25
ml) and filtered. After cooling to room temperature, the mixture
was filtered and the residue recrystallized from 50% aqueous
ethanol (20 ml) to give 0.58 g (53%).
[0066] .sup.1H NMR (200 MHz; CDCl.sub.3: .delta. 2.32 (3H, s),
7.0-7.45 (14H, m).
[0067] .sup.13C NMR (50 MHz; CDCl.sub.3): .delta. 21.1, 57.9,
123.9, 124.5, 127.0, 128.3, 128.4, 128.7, 130.4, 135.6, 139.0,
168.5.
EXAMPLE 11
Preparation of
4-Benzyl-1,2-diphenyl-4-hydroxy-3,5-pyrazolidinedione
[0068] Prepared from 4-benzyl-1,2-diphenyl-3,5-pyrazolidinedione
(3.3 g; 9.6 mmol), 35% H.sub.2O.sub.2 (1.4 ml; 16.3 mmol), and
acetic acid (50 ml) using the procedure of Example 10 to give 1.0 g
(30%).
[0069] .sup.1H NMR (200 MHz; CDCl.sub.3): .delta. 3.30 (2H, s),
6.75-7.3 (15H, m).
[0070] .sup.13C NMR (50 MHz; CDCl.sub.3): .delta. 43.1, 75.4,
123.0, 126.7, 127.5, 128.4, 130.2, 132.1, 134.7, 170.1.
EXAMPLE 12
Antiviral Activity of 4OH--OPB (Example 7)
[0071] 4OH--OPB was added to cultures of growing MT4 cells (a human
CD4 cell line). HIV-1, stored in the culture medium at -75.degree.
C. was thawed and added in an amount which infected about 1 in 7
cells in each culture. The virus was absorbed to the cells for 2.3
hours at ambient temperature whereafter the cultures were
centrifuged at 1200 rpm, the medium was removed, the cells were
suspended in fresh growth medium and 4OH--OPB was added to
concentrations of 1, 10 and 100 .mu.M (diluted in medium from a
stock solution of 20 mM in DMSO). After 72 hours the HIV antigen
concentration was determined using Abbott's test. By way of
comparison phenbutazone (PB) was tested analogously. The results
are shown in FIG. 1 and demonstrate inhibition of virus production
by 4OH--OPB at concentrations above the lowest tested.
EXAMPLE 13
Combination Antiviral Effect with 4OH--OPB
[0072] Cell culture experiments were carried out as in Example 12,
but in place of 4OH--OPB (0-100 .mu.M) was added:
i) 4OH--OPB (0-10 .mu.M) with AZT (0-1 .mu.M) ii) 4OH--OPB (0-100
.mu.M) with Indinavir (0-100 .mu.M) iii) 4OH--OPB (0-100 .mu.M)
with Nevirapin (0-10 .mu.M) iv) 4OH--OPB (0-10 .mu.M) with ddI
(0-100 .mu.M)
[0073] The results are shown in FIGS. 2-5 respectively and
demonstrate the enhanced anti-HIV effect of 4OH--OPB in combination
with other anti-viral agents.
EXAMPLE 14
TABLE-US-00002 [0074] Preparation of capsules for oral use 4-OH OPB
(Example 7) 50 mg Amylum maydis q.s.
[0075] The powder is mixed and filled into hard gelatin capsules
(Capsugel size 00).
EXAMPLE 15
TABLE-US-00003 [0076] Preparation of tablets Gram 4-OH OPB (Example
7) 200 Lactose 85 Polyvinylpyrrolidone 5 Starch 42 Talcum powder 15
Magnesium stearate 3
[0077] 4-OH OPB and lactose are screened through a 0.15 mm sieve
and mixed together with an aqueous solution of
polyvinyl-pyrrolidone. The mass is granulated, and the dried
(40.degree. C.) granulate is mixed with starch, talcum powder and
magnesium stearate. The granulate is compressed into tablets. The
tablet diameter is 11 mm, the tablet weight is 350 mg and each
tablet contains 200 mg 4-OH OPB.
EXAMPLE 16
Preparation of a Suspension for Rectal Administration
[0078] Methyl p-hydroxybeznzoate (70 mg) and
propyl-p-hydroxybenzoate (15 mg) are dissolved in water (100 ml) at
90.degree. C. After cooling to 30.degree. C., methyl cellulose (2
g) is added and the mixture is agitated for 3 hours. 1 gram 4-OH
OPB (Example 7) is screened through a 0.15 mm sieve, and dispersed
in the solution under vigorous stirring. The suspension is filled
in a 100 ml tube. The suspension contains 10 mg 4-OH OPB/ml.
EXAMPLE 17
TABLE-US-00004 [0079] Preparation of oral suspension Gram 4OH OPB
(Example 7) 10 Carboxymethyl cellulose 1.5 Sorbitol 200 Sodium
benzoate 1.0 Orange essence 0.3 Apricot essence 0.7 Ethanol 50
Water 236.5
[0080] Carboxymethyl cellulose, sorbitol and sodium benzoate are
dissolved in water with stirring for 2 hours. A solution of the
essences in ethanol is added. 4-OH OPB is screened through a 0.15
mm sieve and dispersed in the solution under vigorous stirring. The
suspension (10 gram) is filled in a 20 ml tube. Each tube contains
200 mg 4-OH OPB.
EXAMPLE 18
Mouse Toxicity
[0081] 20 g mice were given single doses of 4OH--OPB (20 mM in
DMSO) intraperitoneally. Doses of 1 to 100 .mu.M (in ECF),
corresponding to 0.29 to 29 .mu.M/kg bodyweight, produced no toxic
effect. Furthermore, injection of 4OH--OPB could be increased to
2000 mg/kg (corresponding to 20 mM in the extracellular fluid)
before the mice started to die (6 out of 10 died at 2000 mg/kg).
Thus the concentrations that effectively inhibit HIV replication in
cell cultures are up to 200000 times lower than the lethal dose in
mice.
* * * * *