U.S. patent application number 16/405011 was filed with the patent office on 2019-10-24 for novel combination and use.
This patent application is currently assigned to HELPERBY THERAPEUTICS LIMITED. The applicant listed for this patent is HELPERBY THERAPEUTICS LIMITED. Invention is credited to Anthony COATES, Yanmin HU.
Application Number | 20190321441 16/405011 |
Document ID | / |
Family ID | 55071060 |
Filed Date | 2019-10-24 |
![](/patent/app/20190321441/US20190321441A1-20191024-C00001.png)
![](/patent/app/20190321441/US20190321441A1-20191024-C00002.png)
![](/patent/app/20190321441/US20190321441A1-20191024-C00003.png)
![](/patent/app/20190321441/US20190321441A1-20191024-D00001.png)
![](/patent/app/20190321441/US20190321441A1-20191024-D00002.png)
![](/patent/app/20190321441/US20190321441A1-20191024-D00003.png)
![](/patent/app/20190321441/US20190321441A1-20191024-D00004.png)
![](/patent/app/20190321441/US20190321441A1-20191024-D00005.png)
![](/patent/app/20190321441/US20190321441A1-20191024-D00006.png)
![](/patent/app/20190321441/US20190321441A1-20191024-D00007.png)
![](/patent/app/20190321441/US20190321441A1-20191024-D00008.png)
View All Diagrams
United States Patent
Application |
20190321441 |
Kind Code |
A1 |
COATES; Anthony ; et
al. |
October 24, 2019 |
NOVEL COMBINATION AND USE
Abstract
The present invention relates to the use of one or more
compounds selected from the following: caffeic acid, thymol,
aspirin, benzydamine hydrochloride, diclofenac sodium,
flurbiprofen, ibuprofen, indomethacin, trifluoperazine
hydrochloride, chlorprothixene hydrochloride, triflupromazine
hydrochloride, suloctidil, thioridazine hydrochloride,
dichlorophen, saccharin and piroxicam, in combination with a
polymyxin selected from colistin or polymyxin B or a
pharmaceutically acceptable derivative thereof, for use in the
treatment of a microbial infection, and in particular for killing
clinically latent microorganisms associated with microbial
infections. The invention also provides a combination comprising
suloctidil or a pharmaceutically acceptable derivative or prodrug
thereof, and a polymyxin selected from polymyxin E and polymyxin B
or a pharmaceutically acceptable derivative thereof. This
combination is particularly useful for the treatment and/or
prevention of microbial infections.
Inventors: |
COATES; Anthony; (London,
GB) ; HU; Yanmin; (London, GB) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
HELPERBY THERAPEUTICS LIMITED |
London |
|
GB |
|
|
Assignee: |
HELPERBY THERAPEUTICS
LIMITED
London
GB
|
Family ID: |
55071060 |
Appl. No.: |
16/405011 |
Filed: |
May 7, 2019 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
15537007 |
Jun 16, 2017 |
10335454 |
|
|
PCT/GB2015/054069 |
Dec 18, 2015 |
|
|
|
16405011 |
|
|
|
|
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P 19/08 20180101;
A61K 31/428 20130101; Y02A 50/406 20180101; A61P 31/10 20180101;
Y02A 50/483 20180101; A61P 1/04 20180101; A61P 9/10 20180101; Y02A
50/401 20180101; A61P 1/18 20180101; A61P 9/00 20180101; A61P 43/00
20180101; Y02A 50/481 20180101; A61K 31/382 20130101; A61P 1/12
20180101; A61P 11/02 20180101; A61P 1/02 20180101; A61P 17/02
20180101; A61P 27/16 20180101; A61P 31/04 20180101; A61K 31/405
20130101; A61P 31/08 20180101; A61P 13/08 20180101; A61K 31/5415
20130101; Y02A 50/479 20180101; A61P 29/00 20180101; A61P 15/02
20180101; A61P 11/04 20180101; A61K 31/135 20130101; A61P 31/00
20180101; A61P 39/02 20180101; A61P 1/16 20180101; A61P 15/00
20180101; A61P 27/02 20180101; A61K 31/145 20130101; A61P 13/10
20180101; Y02A 50/473 20180101; A61K 38/12 20130101; A61P 13/02
20180101; Y02A 50/469 20180101; A61K 31/05 20130101; A61P 11/06
20180101; A61P 17/00 20180101; A61K 31/216 20130101; A61K 31/416
20130101; A61K 31/616 20130101; A61P 19/02 20180101; Y02A 50/404
20180101; A61K 31/192 20130101; A61P 11/00 20180101; A61P 31/06
20180101; Y02A 50/30 20180101; A61P 31/02 20180101; A61K 31/196
20130101; A61K 38/12 20130101; A61K 2300/00 20130101; A61K 31/05
20130101; A61K 2300/00 20130101; A61K 31/135 20130101; A61K 2300/00
20130101; A61K 31/216 20130101; A61K 2300/00 20130101; A61K 31/616
20130101; A61K 2300/00 20130101; A61K 31/416 20130101; A61K 2300/00
20130101; A61K 31/196 20130101; A61K 2300/00 20130101; A61K 31/192
20130101; A61K 2300/00 20130101; A61K 31/405 20130101; A61K 2300/00
20130101; A61K 31/5415 20130101; A61K 2300/00 20130101 |
International
Class: |
A61K 38/12 20060101
A61K038/12; A61K 31/145 20060101 A61K031/145; A61K 31/135 20060101
A61K031/135; A61K 31/192 20060101 A61K031/192; A61K 31/196 20060101
A61K031/196; A61K 31/216 20060101 A61K031/216; A61K 31/405 20060101
A61K031/405; A61K 31/416 20060101 A61K031/416; A61K 31/5415
20060101 A61K031/5415; A61K 31/428 20060101 A61K031/428; A61K 31/05
20060101 A61K031/05; A61K 31/382 20060101 A61K031/382; A61K 31/616
20060101 A61K031/616 |
Foreign Application Data
Date |
Code |
Application Number |
Dec 18, 2014 |
GB |
1422670.8 |
Jan 8, 2015 |
GB |
1500278.5 |
Dec 11, 2015 |
GB |
1521901.7 |
Claims
1-17. (canceled)
18. A method of treating a microbial infection, wherein the method
comprises administering to a mammal, one or more compounds selected
from the following: caffeic acid, thymol, aspirin, benzydamine
hydrochloride, diclofenac sodium, flurbiprofen, ibuprofen,
indomethacin, trifluoperazine hydrochloride, chlorprothixene
hydrochloride, triflupromazine hydrochloride, thioridazine
hydrochloride, dichlorophen, saccharin and piroxicam, in
combination with a polymyxin selected from colistin or polymyxin B
or a pharmaceutically acceptable derivative thereof.
19. The method according to claim 18, wherein the method comprises
killing multiplying, non-multiplying and/or clinically latent
microorganisms associated with the microbial infection.
20. The method according to claim 19, wherein the polymyxin is
colistin or a pharmaceutically acceptable derivative thereof.
21. The method according to claim 18, wherein the infection is a
bacterial infection.
22. The method according to claim 21, wherein the bacterial
infection is caused by E. coli, Enterobacteriaceae, Haemophilis
influenzae, Mycobacteria or Klebsiella.
23. The method according to claim 22, wherein the infection is
caused by E. coli or Klebsiella.
24. The method according to claim 23, wherein the infection is
caused by a drug-resistant strain.
25. The method according to claim 24, wherein the infection is
caused by a carbapenemase-resistant strain or "extended spectrum
.beta.-lactamase" (ESPL) strain.
26. The method according to claim 25, wherein the infection is
caused by New Delhi Metallo-beta-lactamase-1 (NDM-1) resistant
Klebs. Pneumonia or NDM-1 E. coli.
27. The method according to claim 18 for the treatment of
tuberculosis, anthrax, abscesses, acne vulgaris, actinomycosis,
asthma, bacilliary dysentry, bacterial conjunctivitis, bacterial
keratitis, bacterial vaginosis, botulism, Buruli ulcer, bone and
joint infections, bronchitis (acute or chronic), brucellosis, burn
wounds, cat scratch fever, cellulitis, chancroid, cholangitis,
cholecystitis, cutaneous diphtheria, cystic fibrosis, cystitis,
diffuse panbronchiolitis, diphtheria, dental caries, diseases of
the upper respiratory tract, eczema, empymea, endocarditis,
endometritis, enteric fever, enteritis, epididymitis, epiglottitis,
erysipelis, erysipclas, erysipeloid, erythrasma, eye infections,
furuncles, gardnerella vaginitis, gastrointestinal infections
(gastroenteritis), genital infections, gingivitis, gonorrhoea,
granuloma inguinale, Haverhill fever, infected burns, infections
following dental operations, infections in the oral region,
infections associated with prostheses, intraabdominal abscesses,
Legionnaire's disease, leprosy, leptospirosis, listeriosis, liver
abscesses, Lyme disease, lymphogranuloma venerium, mastitis,
mastoiditis, meningitis and infections of the nervous system,
mycetoma, nocardiosis, non-specific urethritis, opthalmia,
osteomyelitis, otitis, orchitis, pancreatitis, paronychia,
pelveoperitonitis, peritonitis, peritonitis with appendicitis,
pharyngitis, phlegmons, pinta, plague, pleural effusion, pneumonia,
postoperative wound infections, postoperative gas gangrene,
prostatitis, pseudo-membranous colitis, psittacosis, pulmonary
emphysema, pyelonephritis, pyoderma, Q fever, rat-bite fever,
reticulosis, ricin poisoning, Ritter's disease, salmonellosis,
salpingitis, septic arthritis, septic infections, septicameia,
sinusitis, skin infections, syphilis, systemic infections,
tonsillitis, toxic shock syndrome, trachoma, tularaemia, typhoid,
typhus, urethritis, wound infections, yaws, aspergillosis,
candidiasis, cryptococcosis, favus, histoplasmosis, intertrigo,
mucormycosis, tinea, onychomycosis, pityriasis versicolor, ringworm
and sporotrichosis; or infections with MSSA, MRSA, Staph.
epidermidis, Strept. agalactiae, Strept. pyogenes, Escherichia
coli, Klebs. pneumoniae, Klebs. oxytoca, Pr. mirabilis, Pr.
rettgeri, Pr. vulgaris, Haemophilis influenzae, Enterococcus
faecalis or Enterococcus faecium.
28. A pharmaceutical composition comprising one or more compounds
selected from the following: caffeic acid, thymol, aspirin,
benzydamine hydrochloride, diclofenac sodium, flurbiprofen,
ibuprofen, indomethacin, trifluoperazine hydrochloride,
chlorprothixene hydrochloride, triflupromazine hydrochloride,
suloctidil, thioridazine hydrochloride, dichlorophen, saccharin and
piroxicam, in combination with a polymyxin selected from colistin
or polymyxin B or a pharmaceutically acceptable derivative thereof,
and a pharmaceutically acceptable adjuvant, diluent or carrier.
29. A product comprising one or more compounds selected from the
following: caffeic acid, thymol, aspirin, benzydamine
hydrochloride, diclofenac sodium, flurbiprofen, ibuprofen,
indomethacin, trifluoperazine hydrochloride, chlorprothixene
hydrochloride, triflupromazine hydrochloride, suloctidil,
thioridazine hydrochloride, dichlorophen, saccharin and piroxicam,
in combination with a polymyxin selected from colistin or polymyxin
B or a pharmaceutically acceptable derivative thereof, as a
combined preparation for simultaneous, separate or sequential use
in treating a microbial infection.
30. The product according to claim 29 for killing clinically latent
microorganisms associated with the microbial infection.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application is a Continuation of U.S. application Ser.
No. 15/537,007 filed Jun. 16, 2017, which is a National Stage of
International Application No. PCT/GB2015/054069 filed Dec. 18,
2015, claiming priorities based on British Patent Application Nos.
1422670.8, filed Dec. 18, 2014, 1500278.5, filed Jan. 8, 2015, and
1521901.7, filed Dec. 11, 2015, the contents of all of which are
incorporated herein by reference in their entirety.
BACKGROUND OF THE INVENTION
[0002] The present invention relates to the use of certain known
compounds in combination with an anti-microbial agent for the
treatment of microbial infections. Additionally the present
invention relates to the use of suloctidil or a pharmaceutically
acceptable derivative or prodrug thereof in combination with
polymyxin E or polymyxin B or a pharmaceutically acceptable
derivative thereof for the treatment of microbial infections. In
particular, it relates to the use of such combinations to kill
multiplying and/or clinically latent microorganisms associated with
microbial infections.
[0003] Before the introduction of antibiotics, patients suffering
from acute microbial infections (e.g. tuberculosis or pneumonia)
had a low chance of survival. For example, mortality from
tuberculosis was around 50%. Although the introduction of
antimicrobial agents in the 1940s and 1950s rapidly changed this
picture, bacteria have responded by progressively gaining
resistance to commonly used antibiotics. Now, every country in the
world has antibiotic-resistant bacteria. Indeed, more than 70% of
bacteria that give rise to hospital acquired infections in the USA
resist at least one of the main antimicrobial agents that are
typically used to fight infection (Nature Reviews, Drug Discovery,
1, 895-910 (2002)).
[0004] One way of tackling the growing problem of resistant
bacteria is the development of new classes of antimicrobial agents.
However, until the introduction of linezolid in 2000, there had
been no new class of antibiotic marketed for over 37 years.
Moreover, even the development of new classes of antibiotic
provides only a temporary solution, and indeed there are already
reports of resistance of certain bacteria to linezolid (Lancet,
357, 1179 (2001) and Lancet, 358, 207-208 (2001)).
[0005] In order to develop more long-term solutions to the problem
of bacterial resistance, it is clear that alternative approaches
are required. One such alternative approach is to minimise, as much
as is possible, the opportunities that bacteria are given for
developing resistance to important antibiotics. Thus, strategies
that can be adopted include limiting the use of antibiotics for the
treatment of non-acute infections, as well as controlling which
antibiotics are fed to animals in order to promote growth.
[0006] However, in order to tackle the problem more effectively, it
is necessary to gain an understanding of the actual mechanisms by
which bacteria generate resistance to antibiotic agents. To do this
requires first a consideration of how current antibiotic agents
work to kill bacteria.
[0007] Antimicrobial agents target essential components of
bacterial metabolism. For example, the .beta.-lactams (e.g.
penicillins and cephalosporins) inhibit cell wall synthesis,
whereas other agents inhibit a diverse range of targets, such as
DNA gyrase (quinolones) and protein synthesis (e.g. macrolides,
aminoglycosides, tetracyclines and oxazolidinones). The range of
organisms against which the antimicrobial agents are effective
varies, depending upon which organisms are heavily reliant upon the
metabolic step(s) that is/are inhibited. Further, the effect upon
bacteria can vary from a mere inhibition of growth (i.e. a
bacteriostatic effect, as seen with agents such as the
tetracyclines) to full killing (i.e. a bactericidal effect, as
seen, e.g. with penicillin).
[0008] Bacteria have been growing on Earth for more than 3 billion
years and, in that time, have needed to respond to vast numbers of
environmental stresses. It is therefore perhaps not surprising that
bacteria have developed a seemingly inexhaustible variety of
mechanisms by which they can respond to the metabolic stresses
imposed upon them by antibiotic agents. Indeed, mechanisms by which
the bacteria can generate resistance include strategies as diverse
as inactivation of the drug, modification of the site of action,
modification of the permeability of the cell wall, overproduction
of the target enzyme and bypass of the inhibited steps.
Nevertheless, the rate of resistance emerges to a particular agent
has been observed to vary widely, depending upon factors such as
the agent's mechanism of action, whether the agent's mode of
killing is time- or concentration-dependent, the potency against
the population of bacteria and the magnitude and duration of the
available serum concentration.
[0009] It has been proposed (Science, 264, 388-393 (1994)) that
agents that target single enzymes (e.g. rifampicin) are the most
prone to the development of resistance. Further, the longer that
suboptimal levels of antimicrobial agent are in contact with the
bacteria, the more likely the emergence of resistance.
[0010] Moreover, it is now known that many microbial infections
include sub-populations of bacteria that are phenotypically
resistant to antimicrobials (J. Antimicrob. Chemother., 4, 395-404
(1988); J. Med. Microbiol., 38, 197-202 (1993); J. Bacteriol., 182,
1794-1801 (2000); ibid. 182, 6358-6365 (2000); ibid. 183, 6746-6751
(2001); FEMS Microbiol. Lett., 202, 59-65 (2001); and Trends in
Microbiology, 13, 34-40 (2005)). There appear to be several types
of such phenotypically resistant bacteria, including persisters,
stationary-phase bacteria, as well as those in the depths of
biofilms. However, each of these types is characterised by its low
rate of growth compared to log-phase bacteria under the same
conditions. Nutritional starvation and high cell densities are also
common characteristics of such bacteria.
[0011] Although resistant to antimicrobial agents in their
slow-growing state, phenotypically resistant bacteria differ from
those that are genotypically resistant in that they regain their
susceptibility to antimicrobials when they return to a fast-growing
state (e.g. when nutrients become more readily available to
them).
[0012] The presence of phenotypically resistant bacteria in an
infection leads to the need for prolonged courses of antimicrobial
agents, comprising multiple doses. This is because the resistant,
slowly multiplying bacteria provide a pool of "latent" organisms
that can convert to a fast-growing state when the conditions allow
(thereby effectively re-initiating the infection). Multiple doses
over time deal with this issue by gradually killing off the
"latent" bacteria that convert to "active" form.
[0013] However, dealing with "latent" bacteria by administering
prolonged courses of antimicrobials poses its own problems. That
is, prolonged exposure of bacteria to suboptimal concentrations of
antimicrobial agent can lead to the emergence of genotypically
resistant bacteria, which can then multiply rapidly in the presence
of even high concentrations of the antimicrobial.
[0014] Long courses of antimicrobials are more likely to encourage
the emergence of genotypic resistance than shorter courses on the
grounds that non-multiplying bacterial will tend to survive and,
interestingly, probably have an enhanced ability to mutate to
resistance (Proc. Natl. Acad. Sci. USA, 92, 11736-11740 (1995); J.
Bacteriol., 179, 6688-6691 (1997); and Antimicrob. Agents
Chemother., 44, 1771-1777 (2000)).
[0015] In the light of the above, a new approach to combating the
problem of bacterial resistance might be to select and develop
antimicrobial agents on the basis of their ability to kill "latent"
microorganisms. The production of such agents would allow, amongst
other things, for the shortening of chemotherapy regimes in the
treatment of microbial infections, thus reducing the frequency with
which genotypical resistance arises in microorganisms.
[0016] Recently, there has been report of an anti-retroviral drug,
zidovudine being active as an anti-microbial when combined with
gentamicin. Thus, Doleans-Jordheim A. et al., disclosed (Eur J Clin
Microbiol Infect Dis. 2011 October; 30(10):1249-56) that Zidovudine
(AZT) had a bactericidal effect on some enterobacteria, yet could
induce resistance in Escherichia coli. These resistances were
associated with various modifications in the thymidine kinase gene.
Furthermore, an additive or synergistic activity between AZT and
the two aminoglycoside antibiotics amikacin and gentamicin was
observed against enterobacteria.
[0017] International Patent Application, Publication Number
WO2012032360 discloses that certain classes of biologically active
compounds possess bactericidal activity. One of these classes is
vasodilators including compounds such as perhexiline maleate,
suloctidil or nisoldipine.
[0018] International Patent Application published as WO2014/147405
describes the use of zidovudine in combination with a polymyxin
selected from colistin and polymyxin B for treating a microbial
infection.
[0019] Polymyxins are antibiotic compounds with a general structure
consisting of a cyclic peptide and a long hydrophobic tail. They
are known to disrupt the structure of the bacterial cell membrane
by interacting with its phospholipids, and polymyxins B and E are
typically used in the treatment of Gram-negative bacterial
infections. Polymyxin E is otherwise known as "Colistin", and is
commercially available in Europe under the trade name
Colomycin.RTM. in tablet form. Colomycin.RTM. tablets include the
sulphate salt of colistin and are indicated for the treatment of
gastrointestinal infections caused by sensitive Gram negative
organisms, as well as for bowel preparation. Polymyxin B is
commercially available in Europe under the trade name Maxitrol.RTM.
in the form of eye drops. Maxitrol.RTM. eye drops include polymyxin
B in form of the sulphate salt, and are indicated for the short
term treatment of steroid responsive conditions of the eye when
prophylactic antibiotic treatment is also required, after excluding
the presence of fungal and viral disease.
[0020] Given the importance of antimicrobial agents such as
polymyxins in the fight against bacterial infection, the
identification of further agents capable of enhancing their
anti-bacterial activity addresses an important need.
BRIEF SUMMARY OF THE INVENTION
[0021] The present invention is thus based on the unexpected
finding that the combinations and in particular the combination of
suloctidil and colistin exhibit synergistic antimicrobial activity
against log phase (i.e. multiplying) and/or clinically latent
microorganisms, i.e. greater than the expected additive effect of
each agent at the stated dosage level. The surprising biological
activity of the combinations of the present invention offers the
opportunity to shorten chemotherapy regimens and may result in a
reduction in the emergence of microbial resistance associated with
the use of such combinations.
[0022] Synergy in the context of antimicrobial drugs is measured in
a number of ways that conform to the generally accepted opinion
that "synergy is an effect greater than additive". One of the ways
to assess whether synergy has been observed is to use the
"chequerboard" technique. This is a well-accepted method that leads
to the generation of a value called the fractional inhibitory
concentration index (FICI). Orhan et al J. Clin. Microbiol. 2005,
43(1):140 describes the chequerboard method and analysis in the
paragraph bridging pages 140-141, and explains that the FICI value
is a ratio of the sum of the MIC (Minimum Inhibitory Concentration)
level of each individual component alone and in the mixture. The
combination is considered synergistic when the .SIGMA.FIC is
<0.5, indifferent when the .SIGMA.FIC is >0.5 to <4.0, and
antagonistic when the .SIGMA.FIC is >4.0.
[0023] Another accepted test for ascertaining the presence or
absence of synergy is to use time-kill methods where the dynamic
effect of a drug combination is compared to each drug alone when
assessing the effect on bacterial log or stationary-growth over
time. Again, the possible results are for synergistic, additive or
antagonistic effects.
BRIEF DESCRIPTION OF THE DRAWINGS
[0024] The patent or application file contains at least one drawing
executed in color. Copies of this patent or patent application
publication with color drawing(s) will be provided by the Office
upon request and payment of the necessary fee.
[0025] FIG. 1 is the time kill curve showing combination of
colistin and HT013015 against NDM-1 K. pneumoniae.
[0026] FIG. 2 is the time kill curve showing combination of
colistin and HT0120566 against NDM-1 Klebsiella pneumoniae.
[0027] FIG. 3 is the time kill curve showing combination of
colistin and HT0120707 against NDM-1 E. coli.
[0028] FIG. 4 is the time kill curve showing combination of
colistin and HT0120700 against NDM-1 E. coli.
[0029] FIG. 5 is the time kill curve showing combination of
colistin and HT0120093 against NDM-1 E. coli.
[0030] FIG. 6 is the time kill curve showing combination of
colistin and HT0120553 against NDM-1 Klebsiella pneumoniae.
[0031] FIG. 7 is the time kill curve showing combination of
colistin and HT0121567 against NDM-1 K.pneumoniae.
[0032] FIG. 8 is the time kill curve showing combination of
colistin and HT0121219 against NDM-1 K.pneumoniae
[0033] FIGS. 9 and 10 are the time kill curves showing combination
of colistin and HT0120451 against NDM-1 K.pneumoniae.
[0034] FIG. 11 is the time kill curve showing combination of
colistin and HT0120492 against NDM-1 E. coli.
[0035] FIG. 12 is the time kill curve showing combination of
colistin and HT0120969 against NDM-1 E. coli.
[0036] FIG. 13 is the time kill curve showing combination of
colistin and HT0120448 against NDM-1 Klebsiella pneumoniae.
[0037] FIG. 14 is the time kill curve showing combination of
colistin and HT0120365 against NDM-1 E. coli.
[0038] FIG. 15 is the time kill curve showing combination of
colistin and HT0120417 against NDM-1 E. coli
[0039] FIG. 16 is the time kill curve showing combination of
colistin and HT013001 against NDM-1 E. coli
[0040] FIG. 17 is the time kill curve showing combination of
colistin and HT0120098 against NDM-1 E. coli.
[0041] FIGS. 18-27 are time kill curves showing the combinations
listed in Table 1. Suloctidil is referenced as "93".
[0042] FIGS. 28-34 are time kill curves showing the combinations
listed in Table 2. Suloctidil is referenced as "93".
[0043] FIGS. 35-40 are each a chequerboard showing synergy between
colistin and each of HT013015 (thymol), HT0121219 (aspirin),
HT0120448 (ibuprofen), HT0120451 (indomethacin), HT0120566
(trifluoperazine hydrochloride), and HT0121567 (dichlorophen)
against log phase NDM-1 Klebsiella pneumonia, respectively.
[0044] FIGS. 41-49 are each a chequerboard showing synergy between
colistin and each of HT0120969 (benzydamine hydrochloride),
HT0120365 (diclofenac sodium), HT0120417 (flurbiprofen), HT0120707
(chlorprothixene hydrochloride), HT0120700 (triflupromazine
hydrochloride), HT0120093 (suloctidil), HT0120492 (piroxicam) and
HT0120098 (saccharin) against log phase NDM-1 E. coli,
respectively.
DETAILED DESCRIPTION OF THE INVENTION
[0045] Thus, in one embodiment, the present invention provides a
combination comprising suloctidil or a pharmaceutically acceptable
derivative or prodrug thereof, and a polymyxin selected from
polymyxin E and polymyxin B or a pharmaceutically acceptable
derivative thereof. Preferably the polymyxin is polymyxin E or a
pharmaceutically acceptable derivative thereof.
[0046] Additionally there is provided a pharmaceutical composition
comprising suloctidil or a pharmaceutically acceptable derivative
or prodrug thereof in combination with a polymyxin selected from
polymyxin E and polymyxin B or a pharmaceutically acceptable
derivative thereof, and a pharmaceutically acceptable adjuvant,
diluent or carrier. Preferably the pharmaceutical composition is
for use in the treatment of a microbial infection.
[0047] In a further embodiment the present invention provides the
use of suloctidil or a pharmaceutically acceptable derivative or
prodrug thereof in combination with a polymyxin selected from
polymyxin E and polymyxin B or a pharmaceutically acceptable
derivative thereof, for the manufacture of a medicament for the
prevention and/or treatment of a microbial infection.
[0048] In a further embodiment, the invention provides a method of
preventing or treating a microbial infection which comprises
administering to a mammal, including man, suloctidil or a
pharmaceutically acceptable derivative or prodrug thereof in
combination with a polymyxin selected from polymyxin E and
polymyxin B or a pharmaceutically acceptable derivative
thereof.
[0049] In another embodiment, the invention provides the use of
suloctidil or a pharmaceutically acceptable derivative or prodrug
thereof in combination with a polymyxin selected from polymyxin E
and polymyxin B or a pharmaceutically acceptable derivative thereof
for the prevention and/or treatment of a microbial infection.
[0050] Additionally the present invention provides the use of one
or more compounds selected from the following: caffeic acid,
thymol, aspirin, benzydamine hydrochloride, diclofenac sodium,
flurbiprofen, ibuprofen, indomethacin, trifluoperazine
hydrochloride, chlorprothixene hydrochloride, triflupromazine
hydrochloride, suloctidil, thioridazine hydrochloride,
dichlorophen, saccharin and piroxicam in combination with a
polymyxin selected from colistin or polymyxin B or a
pharmaceutically acceptable derivative thereof, for treating a
microbial infection.
[0051] In a further embodiment, the invention provides a method of
treating a microbial infection which comprises administering to a
mammal, including man, one or more compounds selected from the
following: caffeic acid, thymol, aspirin, benzydamine
hydrochloride, diclofenac sodium, flurbiprofen, ibuprofen,
indomethacin, trifluoperazine hydrochloride, chlorprothixene
hydrochloride, triflupromazine hydrochloride suloctidil,
thioridazine hydrochloride, dichlorophen, saccharin and piroxicam
in combination with a polymyxin selected from colistin or polymyxin
B or a pharmaceutically acceptable derivative thereof.
[0052] There is also provided a pharmaceutical composition
comprising one or more compounds selected from the following:
caffeic acid, thymol, aspirin, benzydamine hydrochloride,
diclofenac sodium, flurbiprofen, ibuprofen, indomethacin,
trifluoperazine hydrochloride, chlorprothixene hydrochloride,
triflupromazine hydrochloride suloctidil, thioridazine
hydrochloride, dichlorophen, saccharin and piroxicam, in
combination with a polymyxin selected from colistin and polymyxin B
or a pharmaceutically acceptable derivative thereof, and a
pharmaceutically acceptable adjuvant, diluent or carrier, for use
in the treatment of a microbial infection, preferably the microbial
infection is a bacterial infection.
[0053] In a further embodiment, the invention relates to a product
comprising one or more compounds selected from: caffeic acid
(hydroxycinnamic acid), thymol, aspirin, benzydamine hydrochloride,
diclofenac sodium, flurbiprofen, ibuprofen, indomethacin,
trifluoperazine hydrochloride, chlorprothixene hydrochloride,
triflupromazine hydrochloride suloctidil, thioridazine
hydrochloride, dichlorophen, saccharin and piroxicam, in
combination with a polymyxin selected from colistin and polymyxin B
or a pharmaceutically acceptable derivative thereof, as a combined
preparation for simultaneous, separate or sequential use in killing
clinically latent microorganism associated with a microbial
infection.
[0054] According to a further embodiment of the invention, there is
provided a product comprising suloctidil or a pharmaceutically
acceptable derivative or prodrug thereof, and a polymyxin selected
from polymyxin E and polymyxin B or a pharmaceutically acceptable
derivative thereof as a combined preparation for simultaneous,
separate or sequential use in the prevention and/or treatment of a
microbial infection.
[0055] The present invention therefore relates to:
[0056] the use of caffeic acid for the treatment of a microbial
infection in combination with colistin;
[0057] the use of thymol for the treatment of a microbial infection
in combination with colistin;
[0058] the use of aspirin for the treatment of a microbial
infection in combination with colistin;
[0059] the use of benzydamine hydrochloride for the treatment of a
microbial infection in combination with colistin;
[0060] the use of diclofenac sodium for the treatment of a
microbial infection in combination with colistin;
[0061] the use of flurbiprofen for the treatment of a microbial
infection in combination with colistin;
[0062] the use of ibuprofen for the treatment of a microbial
infection in combination with colistin;
[0063] the use of indomethacin for the treatment of a microbial
infection in combination with colistin;
[0064] the use of trifluoperazine hydrochloride for the treatment
of a microbial infection in combination with colistin;
[0065] the use of chlorprothixene hydrochloride for the treatment
of a microbial infection in combination with colistin;
[0066] the use of triflupromazine hydrochloride for the treatment
of a microbial infection in combination with colistin;
[0067] the use of suloctidil or a pharmaceutically acceptable
derivative or prodrug thereof for the treatment of a microbial
infection in combination with colistin;
[0068] a combination comprising suloctidil or a pharmaceutically
acceptable derivative or prodrug thereof and colistin or a
pharmaceutically acceptable derivative thereof;
[0069] the use of thioridazine hydrochloride for the treatment of a
microbial infection in combination with colistin;
[0070] the use of dichlorophen for the treatment of a microbial
infection in combination with colistin;
[0071] the use of saccharin for the treatment of a microbial
infection in combination with colistin; and
[0072] the use of piroxicam for the treatment of a microbial
infection in combination with colistin.
[0073] Colistin can also be included in the above combinations in
the form of a pharmaceutically acceptable derivative as defined
herein, e.g. as a salt such as colistin sulphate.
[0074] As described below, the combinations of the present
invention have been demonstrated to be particularly effective
against drug-resistant bacteria, particularly Gram-negative
bacteria, opening the way for said combinations to be administered
both to drug-resistant strains and in said strains before
drug-resistance is built up, i.e. as a first line treatment.
[0075] As used herein, the term "in combination with" covers both
separate and sequential administration of the compound and the
polymyxin. When the compound and polymyxin are administered
sequentially, either the compound or the polymyxin may be
administered first. When administration is simultaneous, the
compound and polymyxin may be administered either in the same or a
different pharmaceutical composition. Adjunctive therapy, i.e.
where one agent is used as a primary treatment and the other agent
is used to assist that primary treatment, is also an embodiment of
the present invention.
[0076] The combinations of the present invention may be used to
treat microbial infections. In particular they may be used to kill
multiplying and/or clinically latent microorganisms associated with
microbial infections. References herein to the treatment of a
microbial infection therefore include killing multiplying and/or
clinically latent microorganisms associated with such infections.
Preferably, the combinations of the present invention are used to
kill clinically latent microorganisms associated with microbial
infections.
[0077] As used herein, "kill" means a loss of viability as assessed
by a lack of metabolic activity.
[0078] As used herein, "clinically latent microorganism" means a
microorganism that is metabolically active but has a growth rate
that is below the threshold of infectious disease expression. The
threshold of infectious disease expression refers to the growth
rate threshold below which symptoms of infectious disease in a host
are absent.
[0079] The metabolic activity of clinically latent microorganisms
can be determined by several methods known to those skilled in the
art; for example, by measuring mRNA levels in the microorganisms or
by determining their rate of uridine uptake. In this respect,
clinically latent microorganisms, when compared to microorganisms
under logarithmic growth conditions (in vitro or in vivo), possess
reduced but still significant levels of: [0080] (I) mRNA (e.g. from
0.0001 to 50%, such as from 1 to 30, 5 to 25 or 10 to 20%, of the
level of mRNA); and/or [0081] (II) uridine (e.g. [.sup.3H]uridine)
uptake (e.g. from 0.0005 to 50%, such as from 1 to 40, 15 to 35 or
20 to 30% of the level of [.sup.3H]uridine uptake).
[0082] Clinically latent microorganisms typically possess a number
of identifiable characteristics. For example, they may be viable
but non-culturable; i.e. they cannot typically be detected by
standard culture techniques, but are detectable and quantifiable by
techniques such as broth dilution counting, microscopy, or
molecular techniques such as polymerase chain reaction. In
addition, clinically latent microorganisms are phenotypically
tolerant, and as such are sensitive (in log phase) to the biostatic
effects of conventional antimicrobial agents (i.e. microorganisms
for which the minimum inhibitory concentration (MIC) of a
conventional antimicrobial is substantially unchanged); but possess
drastically decreased susceptibility to drug-induced killing (e.g.
microorganisms for which, with any given conventional antimicrobial
agent, the ratio of minimum microbiocidal concentration (e.g.
minimum bactericidal concentration, MBC) to MIC is 10 or more).
[0083] As used herein, the term "microorganisms" means fungi and
bacteria. References herein to "microbial", "antimicrobial" and
"antimicrobially" shall be interpreted accordingly. For example,
the term "microbial" means fungal or bacterial, and "microbial
infection" means any fungal or bacterial infection.
[0084] In one embodiment of the invention, one or more of the
aforementioned combinations is used to treat a bacterial infection
in particular, the combinations may be used to kill clinically
latent microorganisms associated with a bacterial infection. As
used herein, the term "bacteria" (and derivatives thereof, such as
"microbial infection") includes, but is not limited to, references
to organisms (or infections due to organisms) of the following
classes and specific types:
[0085] Gram-positive cocci, such as Staphylococci (e.g. Staph.
aureus, Staph. epidermidis, Staph. saprophyticus, Staph.
auricularis, Staph. capitis capitis, Staph. c. ureolyticus, Staph.
caprae, Staph. cohnii cohnii, Staph. c. urealyticus, Staph.
equorum, Staph. gallinarum, Staph. haemolyticus, Staph. hominis
hominis, Staph. h. novobiosepticius, Staph. hyicus, Staph.
intermedius, Staph. lugdunensis, Staph. pasteuri, Staph.
saccharolyticus, Staph. schleiferi schleiferi, Staph. S. coagulans,
Staph. sciuri, Staph. simulans, Staph. warneri and Staph.
xylosus);
[0086] Streptococci (e.g.beta-haemolytic, pyogenic streptococci
(such as Strept. agalactiae, Strept. canis, Strept. dysgalactiae
dysgalactiae, Strept. dysgalactiae equisimilis, Strept. equi equi,
Strept. equi zooepidemicus, Strept. iniae, Strept. porcinus and
Strept. pyogenes),
[0087] microaerophilic, pyogenic streptococci (Streptococcus
"milleri", such as Strept. anginosus, Strept. constellatus
constellatus, Strept. constellatus pharyngidis and Strept.
intermedius), oral streptococci of the "mitis"
(alpha-haemolytic--Streptococcus "viridans", such as Strept. mitis,
Strept. oralis, Strept. sanguinis, Strept. cristatus, Strept.
gordonii and Strept. parasanguinis), "salivarius" (non-haemolytic,
such as Strept. salivarius and Strept. vestibularis) and "mutans"
(tooth-surface streptococci, such as Strept. criceti, Strept.
mutans, Strept. ratti and Strept. sobrinus) groups, Strept.
acidominimus, Strept. bovis, Strept. faecalis, Strept. equinus,
Strept. pneumoniae and Strept. suis, or Streptococci alternatively
classified as Group A, B, C, D, E, G, L, P, U or V
Streptococcus);
[0088] Gram-negative cocci, such as Neisseria gonorrhoeae,
Neisseria meningitidis, Neisseria cinerea, Neisseria elongata,
Neisseria flavescens, Neisseria lactamica, Neisseria mucosa,
Neisseria sicca, Neisseria subflava and Neisseria weaveri;
[0089] Bacillaceae, such as Bacillus anthracis, Bacillus subtilis,
Bacillus thuringiensis, Bacillus stearothermophilus and Bacillus
cereus;
[0090] Enterobacteriaceae, such as Escherichia coli, Enterobacter
(e.g. Enterobacter aerogenes, Enterobacter agglomerans and
Enterobacter cloacae), Citrobacter (such as Citrob. freundii and
Citrob. divernis), Hafnia (e.g. Hafnia alvei), Erwinia (e.g.
Erwinia persicinus), Morganella morganii, Salmonella (Salmonella
enterica and Salmonella typhi), Shigella (e.g. Shigella
dysenteriae, Shigella flexneri, Shigella boydii and Shigella
sonnei), Klebsiella (e.g. Klebs. pneumoniae, Klebs. oxytoca, Klebs.
ornitholytica, Klebs. planticola, Klebs. ozaenae, Klebs. terrigena,
Klebs. granulomatis (Calymmatobacterium granulomatis) and Klebs.
rhinoscleromatis), Proteus (e.g. Pr. mirabilis, Pr. rettgeri and
Pr. vulgaris), Providencia (e.g. Providencia alcalifaciens,
Providencia rettgeri and Providencia stuartii), Serratia (e.g.
Serratia marcescens and Serratia liquifaciens), and Yersinia (e.g.
Yersinia enterocolitica, Yersinia pestis and Yersinia
pseudotuberculosis);
[0091] Enterococci (e.g. Enterococcus avium, Enterococcus
casseliflavus, Enterococcus cecorum, Enterococcus dispar,
Enterococcus durans, Enterococcus faecalis, Enterococcus faecium,
Enterococcus flavescens, Enterococcus gallinarum, Enterococcus
hirae, Enterococcus malodoratus, Enterococcus mundtii, Enterococcus
pseudoavium, Enterococcus raffinosus and Enterococcus
solitarius);
[0092] Helicobacter (e.g. Helicobacter pylori, Helicobacter cinaedi
and Helicobacter fennelliae); Acinetobacter (e.g. A. baumanfi, A.
calcoaceticus, A. haemolyticus, A. johnsonii, A. junfi, A. Iwoffi
and A. radioresistens);
[0093] Pseudomonas (e.g. Ps. aeruginosa, Ps. maltophilia
(Stenotrophomonas maltophilia), Ps. alcaligenes, Ps. chlororaphis,
Ps. fluorescens, Ps. luteola. Ps. mendocina, Ps. monteilii, Ps.
oryzihabitans, Ps. pertocinogena, Ps. pseudalcaligenes, Ps. putida
and Ps. stutzeri); Bacteroides fragilis;
[0094] Peptococcus (e.g. Peptococcus niger);
Peptostreptococcus;
[0095] Clostridium (e.g. C. perfringens, C. difficile, C.
botulinum, C. tetani, C. absonum, C. argentinense, C. baratii, C.
bifermentans, C. beijerinckii, C. butyricum, C. cadaveris, C.
camis, C. celatum, C. clostridioforme, C. cochlearium, C.
cocleatum, C. fallax, C. ghonii, C. glycolicum, C. haemolyticum, C.
hastiforme, C. histolyticum, C. indolis, C. innocuum, C.
irregulare, C. leptum, C. limosum, C. malenominatum, C. novyi, C.
oroticum, C. paraputrificum, C. piliforme, C. putrefasciens, C.
ramosum, C. septicum, C. sordelii, C. sphenoides, C. sporogenes, C.
subterminale, C. symbiosum and C. tertium); Mycoplasma (e.g. M.
pneumoniae, M. hominis, M. genitalium and M. urealyticum);
[0096] Mycobacteria (e.g. Mycobacterium tuberculosis, Mycobacterium
avium, Mycobacterium fortuitum, Mycobacterium marinum,
Mycobacterium kansasii, Mycobacterium chelonae, Mycobacterium
abscessus, Mycobacterium leprae, Mycobacterium smegmitis,
Mycobacterium africanum, Mycobacterium alvei, Mycobacterium
asiaticum, Mycobacterium aurum, Mycobacterium bohemicum,
Mycobacterium bovis, Mycobacterium branderi, Mycobacterium brumae,
Mycobacterium celatum, Mycobacterium chubense, Mycobacterium
confluentis, Mycobacterium conspicuum, Mycobacterium cookii,
Mycobacterium flavescens, Mycobacterium gadium, Mycobacterium
gastri, Mycobacterium genavense, Mycobacterium gordonae,
Mycobacterium goodii, Mycobacterium haemophilum, Mycobacterium
hassicum, Mycobacterium intracellulare, Mycobacterium interjectum,
Mycobacterium heidelberense, Mycobacterium lentiflavum,
Mycobacterium malmoense, Mycobacterium microgenicum, Mycobacterium
microti, Mycobacterium mucogenicum, Mycobacterium neoaurum,
Mycobacterium nonchromogenicum, Mycobacterium peregrinum,
Mycobacterium phlei, Mycobacterium scrofulaceum, Mycobacterium
shimoidei, Mycobacterium simiae, Mycobacterium szulgai,
Mycobacterium terrae, Mycobacterium thermoresistabile,
Mycobacterium triplex, Mycobacterium triviale, Mycobacterium
tusciae, Mycobacterium ulcerans, Mycobacterium vaccae,
Mycobacterium wolinskyi and Mycobacterium xenopi);
[0097] Haemophilus (e.g. Haemophilus influenzae, Haemophilus
ducreyi, Haemophilus aegyptius, Haemophilus parainfluenzae,
Haemophilus haemolyticus and Haemophilus parahaemolyticus);
[0098] Actinobacillus (e.g. Actinobacillus actinomycetemcomitans,
Actinobacillus equuli, Actinobacillus hominis, Actinobacillus
lignieresii, Actinobacillus suis and Actinobacillus ureae);
Actinomyces (e.g. Actinomyces israelii);
[0099] Brucella (e.g. Brucella abortus, Brucella canis, Brucella
melintensis and Brucella suis); Campylobacter (e.g. Campylobacter
jejuni, Campylobacter coli, Campylobacter lari and Campylobacter
fetus); Listeria monocytogenes;
[0100] Vibrio (e.g. Vibrio cholerae and Vibrio parahaemolyticus,
Vibrio alginolyticus, Vibrio carchariae, Vibrio fluvialis, Vibrio
fumissii, Vibrio hollisae, Vibrio metschnikovii, Vibrio mimicus and
Vibrio vulnificus);
[0101] Erysipelothrix rhusopathiae;
[0102] Corynebacteriaceae (e.g. Corynebacterium diphtheriae,
Corynebacterium jeikeum and Corynebacterium urealyticum);
[0103] Spirochaetaceae, such as Borrelia (e.g. Borrelia
recurrentis, Borrelia burgdorferi, Borrelia afzelii, Borrelia
andersonii, Borrelia bissettii, Borrelia garinii, Borrelia
japonica, Borrelia lusitaniae, Borrelia tanukii, Borrelia turdi,
Borrelia valaisiana, Borrelia caucasica, Borrelia crocidurae,
Borrelia duttoni, Borrelia graingeri, Borrelia hermsii, Borrelia
hispanica, Borrelia latyschewii, Borrelia mazzottii, Borrelia
parkeri, Borrelia persica, Borrelia turicatae and Borrelia
venezuelensis) and Treponema (Treponema pallidum ssp. pallidum,
Treponema pallidum ssp. endemicum, Treponema pallidum ssp. pertenue
and Treponema carateum); Pasteurella (e.g. Pasteurella aerogenes,
Pasteurella bettyae, Pasteurella canis, Pasteurella dagmatis,
Pasteurella gallinarum, Pasteurella haemolytica, Pasteurella
multocida multocida, Pasteurella multocida gallicida, Pasteurella
multocida septica, Pasteurella pneumotropica and Pasteurella
stomatis);
[0104] Bordetella (e.g. Bordetella bronchiseptica, Bordetella
hinzii, Bordetella holmseii, Bordetella parapertussis, Bordetella
pertussis and Bordetella trematum);
[0105] Nocardiaceae, such as Nocardia (e.g. Nocardia asteroides and
Nocardia brasiliensis); Rickettsia (e.g. Ricksettsii or Coxiella
burnetii);
[0106] Legionella (e.g. Legionalla anisa, Legionalla
birminghamensis, Legionalla bozemanii, Legionalla cincinnatiensis,
Legionalla dumoffii, Legionalla feeleii, Legionalla gormanii,
Legionalla hackeliae, Legionalla israelensis, Legionalla jordanis,
Legionalla lansingensis, Legionalla longbeachae, Legionalla
maceachernii, Legionalla micdadei, Legionalla oakridgensis,
Legionalla pneumophila, Legionalla sainthelensi, Legionalla
tucsonensis and Legionalla wadsworthii);
[0107] Moraxella catarrhalis;
[0108] Cyclospora cayetanensis;
[0109] Entamoeba histolytica;
[0110] Giardia lamblia;
[0111] Trichomonas vaginalis;
[0112] Toxoplasma gondii;
[0113] Stenotrophomonas maltophilia;
[0114] Burkholderia cepacia; Burkholderia mallei and Burkholderia
pseudomallei;
[0115] Francisella tularensis;
[0116] Gardnerella (e.g. Gardneralla vaginalis and Gardneralla
mobiluncus);
[0117] Streptobacillus moniliformis;
[0118] Flavobacteriaceae, such as Capnocytophaga (e.g.
Capnocytophaga canimorsus, Capnocytophaga cynodegmi, Capnocytophaga
gingivalis, Capnocytophaga granulosa, Capnocytophaga haemolytica,
Capnocytophaga ochracea and Capnocytophaga sputigena); Bartonella
(Bartonella bacilliformis, Bartonella clarridgeiae, Bartonella
elizabethae, Bartonella henselae, Bartonella quintana and
Bartonella vinsonii arupensis);
[0119] Leptospira (e.g. Leptospira biflexa, Leptospira
borgpetersenii, Leptospira inadai, Leptospira interrogans,
Leptospira kirschneri, Leptospira noguchii, Leptospira santarosai
and Leptospira weilii);
[0120] Spirillium (e.g. Spirillum minus);
[0121] Baceteroides (e.g. Bacteroides caccae, Bacteroides
capillosus, Bacteroides coagulans, Bacteroides distasonis,
Bacteroides eggerthii, Bacteroides forsythus, Bacteroides fragilis,
Bacteroides merdae, Bacteroides ovatus, Bacteroides putredinis,
Bacteroides pyogenes, Bacteroides splanchinicus, Bacteroides
stercoris, Bacteroides tectus, Bacteroides thetaiotaomicron,
Bacteroides uniformis, Bacteroides ureolyticus and Bacteroides
vulgatus);
[0122] Prevotella (e.g. Prevotella bivia, Prevotella buccae,
Prevotella corporis, Prevotella dentalis (Mitsuokella dentalis),
Prevotella denticola, Prevotella disiens, Prevotella enoeca,
Prevotella heparinolytica, Prevotella intermedia, Prevotella
loeschii, Prevotella melaninogenica, Prevotella nigrescens,
Prevotella oralis, Prevotella oris, Prevotella oulora, Prevotella
tannerae, Prevotella venoralis and Prevotella zoogleoformans);
[0123] Porphyromonas (e.g. Porphyromonas asaccharolytica,
Porphyromonas cangingivalis, Porphyromonas canoris, Porphyromonas
cansulci, Porphyromonas catoniae, Porphyromonas circumdentaria,
Porphyromonas crevioricanis, Porphyromonas endodontalis,
Porphyromonas gingivalis, Porphyromonas gingivicanis, Porphyromonas
levii and Porphyromonas macacae);
[0124] Fusobacterium (e.g. F. gonadiaformans, F. mortiferum, F.
naviforme, F. necrogenes, F. necrophorum necrophorum, F.
necrophorum fundiliforme, F. nucleatum nucleatum, F. nucleatum
fusiforme, F. nucleatum polymorphum, F. nucleatum vincentii, F.
periodonticum, F. russii, F. ulcerans and F. varium);
[0125] Chlamydia (e.g. Chlamydia trachomatis);
[0126] Cryptosporidium (e.g. C. parvum, C. hominis, C. canis, C.
felis, C. meleagridis and C. muris); Chlamydophila (e.g.
Chlamydophila abortus (Chlamydia psittaci), Chlamydophila
pneumoniae (Chlamydia pneumoniae) and Chlamydophila psittaci
(Chlamydia psittaci)); Leuconostoc (e.g. Leuconostoc citreum,
Leuconostoc cremoris, Leuconostoc dextranicum, Leuconostoc lactis,
Leuconostoc mesenteroides and Leuconostoc pseudomesenteroides);
[0127] Gemella (e.g. Gemella bergeri, Gemella haemolysans, Gemella
morbillorum and Gemella sanguinis); and
[0128] Ureaplasma (e.g. Ureaplasma parvum and Ureaplasma
urealyticum).
[0129] Preferably, the bacterial infections treated by the
combinations described herein are gram-negative infections.
[0130] Particular bacteria that may be treated using a combination
of the invention include Gram negative bacteria:
[0131] Enterobacteriaceae, such as Escherichia coli, Klebsiella
(e.g. Klebs. pneumoniae and Klebs. oxytoca) and Proteus (e.g. Pr.
mirabilis, Pr. rettgeri and Pr. vulgaris);
[0132] Haemophilis influenzae;
[0133] Mycobacteria, such as Mycobacterium tuberculosis.
[0134] Preferably, the bacterium is Enterobacteriaceae, such as
Escherichia coli, Klebsiella (e.g. Klebs. pneumoniae and Klebs.
oxytoca) and Proteus (e.g. Pr. mirabilis, Pr. rettgeri and Pr.
vulgaris). The combination of the present invention is particularly
beneficial in treating (multi)-drug-resistant ((M)DR) bacteria.
With respect to Enterobacteriaceae, drug resistance most often
builds up to carbapenemase i.e. carbapenemase-resistant strains and
"extended spectrum .beta.-lactamase" (ESBL) strains for example New
Delhi Metallo-beta-lactamase-1 (NDM-1) resistant Klebs.
Pneumonia.
[0135] More preferably the bacterium is Escherichia coli or
Klebsiella (e.g. Klebs. pneumoniae).
[0136] It should be kept in mind that although a combination such
as that claimed may initially be demonstrated to be functional in
treating (M)DR strains, they can then be used in treating
non-resistant strains. This is especially valuable in the context
of the presently claimed combination where the primary therapy for
Enterobacteriaceae, such as Escherichia coli, Klebsiella (e.g.
Klebs. pneumoniae and Klebs. oxytoca) and Proteus (e.g. Pr.
mirabilis, Pr. rettgeri and Pr. vulgaris) are anti-microbial drugs
that are expensive due to prevailing patent protection. The
replacement of such "ethical" drugs by a combination of "generic"
antibiotics is thought to be beneficial from a therapeutic
perspective as well as financial/economic perspective in times
where governments are seeking to reduce the cost of healthcare.
[0137] The combinations of the present invention may be used to
treat infections associated with any of the above-mentioned
bacterial organisms, and in particular they may be used for killing
multiplying and/or clinically latent microorganisms associated with
such an infection.
[0138] Particular conditions which may be treated using the
combination of the present invention include tuberculosis (e.g.
pulmonary tuberculosis, non-pulmonary tuberculosis (such as
tuberculosis lymph glands, genito-urinary tuberculosis,
tuberculosis of bone and joints, tuberculosis meningitis) and
miliary tuberculosis), anthrax, abscesses, acne vulgaris,
actinomycosis, asthma, bacillary dysentery, bacterial
conjunctivitis, bacterial keratitis, bacterial vaginosis, botulism,
Buruli ulcer, bone and joint infections, bronchitis (acute or
chronic), brucellosis, burn wounds, cat scratch fever, cellulitis,
chancroid, cholangitis, cholecystitis, cutaneous diphtheria, cystic
fibrosis, cystitis, nephritis, diffuse panbronchiolitis,
diphtheria, dental caries, diseases of the upper respiratory tract,
eczema, empyema, endocarditis, endometritis, enteric fever,
enteritis, epididymitis, epiglottitis, erysipelas, erysipelas,
erysipeloid, erythrasma, eye infections, furuncles, gardnerella
vaginitis, gastrointestinal infections (gastroenteritis), genital
infections, gingivitis, gonorrhoea, granuloma inguinale, Haverhill
fever, infected burns, infections following dental operations,
infections in the oral region, infections associated with
prostheses, intraabdominal abscesses, Legionnaire's disease,
leprosy, leptospirosis, listeriosis, liver abscesses, Lyme disease,
lymphogranuloma venerium, mastitis, mastoiditis, meningitis and
infections of the nervous system, mycetoma, nocardiosis (e.g.
Madura foot), non-specific urethritis, opthalmia (e.g. opthalmia
neonatorum), osteomyelitis, otitis (e.g. otitis externa and otitis
media), orchitis, pancreatitis, paronychia, pelveoperitonitis,
peritonitis, peritonitis with appendicitis, pharyngitis, phlegmons,
pinta, plague, pleural effusion, pneumonia, postoperative wound
infections, postoperative gas gangrene, prostatitis,
pseudo-membranous colitis, psittacosis, pulmonary emphysema,
pyelonephritis, pyoderma (e.g. impetigo), Q fever, rat-bite fever,
reticulosis, ricin poisoning, Ritter's disease, salmonellosis,
salpingitis, septic arthritis, septic infections, septicameia,
sinusitis, skin infections (e.g. skin granulomas, impetigo,
folliculitis and furunculosis), syphilis, systemic infections,
tonsillitis, toxic shock syndrome, trachoma, tularaemia, typhoid,
typhus (e.g. epidemic typhus, murine typhus, scrub typhus and
spotted fever), urethritis, urinary tract infections, wound
infections, yaws, aspergillosis, candidiasis (e.g. oropharyngeal
candidiasis, vaginal candidiasis or balanitis), cryptococcosis,
favus, histoplasmosis, intertrigo, mucormycosis, tinea (e.g. tinea
corporis, tinea capitis, tinea cruris, tinea pedis and tinea
unguium), onychomycosis, pityriasis versicolor, ringworm and
sporotrichosis; or infections with MSSA, MRSA, Staph. epidermidis,
Strept. agalactiae, Strept. pyogenes, Escherichia coli, Klebs.
pneumoniae, Klebs. oxytoca, Pr. mirabilis, Pr. rettgeri, Pr.
vulgaris, Haemophilis influenzae, Enterococcus faecalis and
Enterococcus faecium. In particular, the combination in kidney
stone associated infections and catheter-associated infections
arising from any of the bacteria described.
[0139] It will be appreciated that references herein to "treatment"
extend to prophylaxis as well as the treatment of established
diseases or symptoms.
[0140] As used herein the term "pharmaceutically acceptable
derivative" means:
[0141] (a) pharmaceutically acceptable salts; and/or
[0142] (b) solvates (including hydrates).
[0143] Pharmaceutically acceptable salts and solvates (including
hydrates) are also understood to include polymorphs such as
pseudopolymorphs, packing polymorphs and conformational
polymorphs.
[0144] A review of suitable pharmaceutical salts may be found in
Berge et al, J Pharm Sci, 66, 1-19 (1977) as well as P. H. Stahl
and C. G. Wermuth, editors, Handbook of Pharmaceutical Salts:
Properties, Selection and Use, Weinheim/Zurich:Wiley-VCH/VHCA,
2002. Suitable acid addition salts include carboxylate salts (e.g.
formate, acetate, trifluoroacetate, propionate, isobutyrate,
heptanoate, decanoate, caprate, caprylate, stearate, acrylate,
caproate, propiolate, ascorbate, citrate, glucuronate, glutamate,
glycolate, .alpha.-hydroxybutyrate, lactate, tartrate,
phenylacetate, mandelate, phenylpropionate, phenylbutyrate,
benzoate, chlorobenzoate, methylbenzoate, hydroxybenzoate,
methoxybenzoate, dinitrobenzoate, o-acetoxybenzoate, salicylate,
nicotinate, isonicotinate, cinnamate, oxalate, malonate, succinate,
suberate, sebacate, fumarate, malate, maleate, hydroxymaleate,
hippurate, phthalate or terephthalate salts), halide salts (e.g.
chloride, bromide or iodide salts), sulfonate salts (e.g.
benzenesulfonate, methyl-, bromo- or chloro-benzenesulfonate,
xylenesulfonate, methanesulfonate, ethanesulfonate,
propanesulfonate, hydroxyethanesulfonate, 1- or
2-naphthalene-sulfonate or 1,5-naphthalenedisulfonate salts) or
sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate,
monohydrogenphosphate, dihydrogenphosphate, metaphosphate,
pyrophosphate or nitrate salts, and the like.
[0145] The antibacterial agent is preferably a polymyxin such as
colistin and/or polymyxin B or a pharmaceutically acceptable
derivative thereof. Particularly preferred is colistin or a
pharmaceutically acceptable derivative thereof. A preferred salt of
colistin is the sulfate salt thereof, i.e. polymyxin B sulfate. A
preferred salt of polymyxin E is the sulfate salt thereof, i.e.
polymyxin E sulfate.
[0146] Several of the compounds included in the combinations of the
present invention including suloctidil, polymyxin B, colistin
sulfate, polymyxin E sulfate and colistin are commercially
available, for example from Sigma Aldrich Limited. Others may be
prepared using conventional methods known in the art.
[0147] The invention further includes suloctidil in prodrug form,
i.e. in the form of a covalently bonded compound which releases the
active suloctidil in vivo. Suloctidil has the following
formula:
##STR00001##
[0148] Such prodrugs are generally the active ingredient, i.e.
suloctidil, wherein one or more appropriate groups (typically the
OH or NH group) have been modified such that the modification may
be reversed upon administration to a human or mammalian subject.
Reversion is usually performed by an enzyme naturally present in
such subject, though it is possible for a second agent to be
administered together with such a prodrug in order to perform the
reversion in vivo. Examples of such modifications to suloctidil
include 1-N-substituted (acyloxy)-alkyl carbamate compounds and
esters. With an ester prodrug the reversion to suloctidil may be
carried out by an esterase.
[0149] Esters are typically formed using organic acids. Organic
acids that may be used include carboxylic acids, such as
alkanecarboxylic acids of 1 to 12 carbon atoms which are
unsubstituted or substituted (e.g., by halogen), such as acetic
acid; with saturated or unsaturated dicarboxylic acid, for example
oxalic, malonic, succinic, maleic, fumaric, phthalic or
tetraphthalic; with hydroxycarboxylic acids, for example ascorbic,
glycolic, lactic, malic, tartaric or citric acid; with amino acids,
for example aspartic or glutamic acid; with benzoic acid; or with
organic sulfonic acids, such as (C.sub.1-C.sub.4)-alkyl- or
aryl-sulfonic acids which are unsubstituted or substituted (for
example, by a halogen) such as methane- or p-toluene sulfonic acid.
In some cases it may be desirable to prepare double ester type
prodrugs such as (acyloxy)alkyl esters or
((alkoxycarbonyl)oxy)alkyl esters).
[0150] In one embodiment the prodrug of suloctidil is a compound of
formula (1):
##STR00002##
[0151] wherein:
[0152] R is H or COR.sup.a where R.sup.a is hydrogen, an optionally
substituted alkyl group, an optionally substituted alkenyl group,
an optionally substituted alkynyl group, an optionally substituted
aryl group, an optionally substituted heteroaryl group, an
optionally substituted carbocyclyl group, an optionally substituted
heterocyclyl group, or an alkoxy group.
[0153] Representative COR.sup.a groups include, but are not limited
to, formyl (--CHO), acetyl, (--C(O)CH.sub.3), cyclohexylcarbonyl,
cyclohexylmethylcarbonyl, benzoyl (C(O)Ph), benzylcarbonyl
(C(O)CH.sub.2Ph), C(O)--C.sub.1-8alkyl,
C(O)(CH.sub.2).sub.t(C.sub.6-C.sub.10aryl),
C(O)(CH.sub.2).sub.t(5-10 membered heteroaryl),
C(O)(CH.sub.2).sub.t(C.sub.3-C.sub.10cycloalkyl), and
C(O)(CH.sub.2).sub.t(4-10 membered heterocyclyl), wherein t is an
integer from 0 to 4.
[0154] R.sup.a is preferably selected from C.sub.1-C.sub.8 alkyl or
C.sub.1-C.sub.4 alkyl, each optionally substituted with halo, OH,
OR.sup.b, or NHSO.sub.2R.sup.b wherein R.sup.b is C.sub.1-4 alkyl;
or C.sub.3-C.sub.10cycloalkyl, 4-10 membered heterocyclyl,
C.sub.6-C.sub.10aryl, arylalkyl, 5-10 membered heteroaryl or
heteroarylalkyl, each optionally substituted with C.sub.1-4 alkyl,
C.sub.1-4 hydroxyalkyl, C.sub.1-4 haloalkoxy, OH, OR.sup.b, or
NHSO.sub.2R.sup.b wherein R.sup.b is C.sub.1-4 alkyl.
[0155] and wherein:
[0156] R.sup.1 is H, or COCR.sup.cR.sup.dOC(O)R.sup.e, wherein
R.sup.c is H or an optionally substituted alkyl group, an
optionally substituted alkenyl group, an optionally substituted
alkynyl group, or an alkoxy group.
[0157] R.sup.c is preferably H or a group selected from
C.sub.1-C.sub.8 alkyl or C.sub.1-C.sub.4 alkyl, each optionally
substituted with halo, OH, OR.sup.b, or NHSO.sub.2R.sup.b wherein
R.sup.b is C.sub.1-4 alkyl, C.sub.3-C.sub.10cycloalkyl, 4-10
membered heterocyclyl, C.sub.6-C.sub.10aryl, 5-10 membered
heteroaryl, wherein the C.sub.3-C.sub.10cycloalkyl, 4-10 membered
heterocyclyl, C.sub.6-C.sub.10aryl, and 5-10 membered heteroaryl
are each optionally substituted with C.sub.1-4 alkyl, C.sub.1-4
hydroxyalkyl, C.sub.1-4 haloalkoxy, OH, OR.sup.b, or
NHSO.sub.2R.sup.b wherein R.sup.b is C.sub.1-4 alkyl.
[0158] More preferably R.sup.c is H or a group selected from
C.sub.1-C.sub.8 alkyl or C.sub.1-C.sub.4 alkyl, each optionally
substituted with halo, OH, OR.sup.b, or NHSO.sub.2R.sup.b wherein
R.sup.b is C.sub.1-4 alkyl. Most preferably R.sup.c is H or an
unsubstituted C.sub.1-4 alkyl.
[0159] R.sup.d is an optionally substituted alkyl group, an
optionally substituted alkenyl group, an optionally substituted
alkynyl group, or an alkoxy group.
[0160] R.sup.d is preferably a group selected from C.sub.1-C.sub.8
alkyl or C.sub.1-C.sub.4 alkyl, each optionally substituted with
halo, OH, OR.sup.b, or NHSO.sub.2R.sup.b wherein R.sup.b is
C.sub.1-4 alkyl, C.sub.3-C.sub.10cycloalkyl, 4-10 membered
heterocyclyl, C.sub.6-C.sub.10aryl, 5-10 membered heteroaryl,
wherein the C.sub.3-C.sub.10cycloalkyl, 4-10 membered heterocyclyl,
C.sub.6-C.sub.10aryl, and 5-10 membered heteroaryl are each
optionally substituted with C.sub.1-4 alkyl, C.sub.1-4
hydroxyalkyl, C.sub.1-4 haloalkoxy, OH, OR.sup.b, or
NHSO.sub.2R.sup.b wherein R.sup.b is C.sub.1-4 alkyl.
[0161] More preferably R.sup.d is selected from C.sub.1-C.sub.8
alkyl or C.sub.1-C.sub.4 alkyl, each optionally substituted with
halo, OH, OR.sup.b, or NHSO.sub.2R.sup.b wherein R.sup.b is
C.sub.1-4 alkyl.
[0162] R.sup.e is an optionally substituted alkyl group, an
optionally substituted alkenyl group, an optionally substituted
alkynyl group, an optionally substituted aryl group, an optionally
substituted heteroaryl group, an optionally substituted carbocyclyl
group, an optionally substituted heterocyclyl group, or an alkoxy
group.
[0163] Representative COR.sup.e groups include, but are not limited
to, formyl (--CHO), acetyl, (--C(O)CH.sub.3), cyclohexylcarbonyl,
cyclohexylmethylcarbonyl, benzoyl (C(O)Ph), benzylcarbonyl
(C(O)CH.sub.2Ph), C(O)--C.sub.1-8alkyl,
C(O)(CH.sub.2).sub.t(C.sub.6-C.sub.10aryl),
C(O)(CH.sub.2).sub.t(5-10 membered heteroaryl),
C(O)(CH.sub.2).sub.t(C.sub.3-C.sub.10cycloalkyl), and
C(O)(CH.sub.2).sub.t(4-10 membered heterocycyl), wherein t is an
integer from 0 to 4.
[0164] Preferably R.sup.e is a group selected from C.sub.1-C.sub.8
alkyl or C.sub.1-C.sub.4 alkyl, each optionally substituted with
halo, OH, OR.sup.b, or NHSO.sub.2R.sup.b wherein R.sup.b is
C.sub.1-4 alkyl, C.sub.3-C.sub.10cycloalkyl, 4-10 membered
heterocyclyl, C.sub.6-C.sub.10aryl, 5-10 membered heteroaryl,
wherein the C.sub.3-C.sub.10cycloalkyl, 4-10 membered heterocyclyl,
C.sub.6-C.sub.10aryl, and 5-10 membered heteroaryl are each
optionally substituted with C.sub.1-4 alkyl, C.sub.1-4
hydroxyalkyl, C.sub.1-4 haloalkoxy, OH, OR.sup.b, or
NHSO.sub.2R.sup.b wherein R.sup.b is C.sub.1-4 alkyl; or
C.sub.3-C.sub.10cycloalkyl, 4-10 membered heterocyclyl,
C.sub.6-C.sub.10aryl, arylalkyl, 5-10 membered heteroaryl or
heteroarylalkyl, each optionally substituted with C.sub.1-4 alkyl,
C.sub.1-4 hydroxyalkyl, C.sub.1-4 haloalkoxy, OH, OR.sup.b, or
NHSO.sub.2R.sup.b wherein R.sup.b is C.sub.1-4 alkyl.
[0165] More preferably R.sup.d and R.sup.e are independently an
unsubstituted C.sub.1-4 alkyl group.
[0166] Provided that if R is H, R.sup.1 is not H, and when R.sup.1
is H, R is not H.
[0167] Alternatively R.sup.1 is a group of formula (2) linked via
the bond indicated, wherein R.sup.f is an optionally substituted
C.sub.1-4 alkyl group, aryl or alkoxyaryl. Preferably R.sup.f is an
unsubstituted C.sub.1-4 alkyl, phenyl or methoxyphenyl.
##STR00003##
[0168] Compounds of formula (1) and (2) can be prepared by known
methods by those skilled in the art. Suitable methods are disclosed
in for example WO 2014/134005, or J. Med. Chem 1996, 39, 480. These
methods are incorporated herein by reference.
[0169] Other prodrug systems will be well known to those skilled in
the art.
[0170] As used herein, the term "alkyl" includes both saturated
straight chain and branched alkyl groups which may be substituted
(mono- or poly-) or unsubstituted. Preferably, the alkyl group is a
C.sub.1-15 alkyl group, more preferably a C.sub.1-10 alkyl group,
more preferably still a C.sub.1-8 alkyl group, more preferably
still a C.sub.1-6 alkyl group. Particularly preferred alkyl groups
include, for example, methyl, ethyl, n-propyl, iso-propyl, n-butyl,
sec-butyl, tert-butyl, n-pentyl, tert-pentyl, neo-pentyl,
iso-pentyl, sec-pentyl and 4-pentyl. In certain embodiments the
alkyl group is substituted with halo, OH, OR.sup.b,
NHSO.sub.2R.sup.b wherein R.sup.b is C.sub.1-4 alkyl.
[0171] The term "halo" refers to fluoro, chloro, bromo or iodo.
[0172] As used herein, the term "aryl" refers to a C.sub.6-18
aromatic group which may be substituted (mono- or poly-) or
unsubstituted. Preferably the aryl group is a C.sub.6-14 aryl
group, more preferably a C.sub.6-10 aryl group. Typical examples
include phenyl, naphthyl, mesityl, benzyl, and anthracenyl, and a
particularly preferred aryl group is phenyl, mesityl or benzyl,
e.g. phenyl.
[0173] As used herein, the term "alkenyl" refers to a carbon chain
containing one or more carbon-carbon double bonds, which may be
branched or unbranched, and substituted (mono- or poly-) or
unsubstituted. Preferably the alkenyl group is a C.sub.2-20 alkenyl
group, more preferably a C.sub.2-15 alkenyl group, more preferably
still a C.sub.2-10 alkenyl group, more preferably still a C.sub.2-8
alkenyl group, or more preferably still a C.sub.2-6 alkenyl
group.
[0174] As used herein, the term "alkynyl" refers to a carbon chain
containing one or more carbon-carbon triple bonds, which may be
branched or unbranched, and substituted (mono- or poly-) or
unsubstituted. Preferably the alkynyl group is a C.sub.2-20 alkynyl
group, more preferably a C.sub.2-15 alkynyl group, more preferably
still a C.sub.2-10 alkynyl group, more preferably still a C.sub.2-8
alkynyl group, or more preferably still a C.sub.2-6 alkynyl
group.
[0175] As used herein, the term "cycloalkyl" refers to a mono- or
multi-ringed cyclic alkyl group which may be substituted (mono- or
poly-) or unsubstituted. Preferably the cycloalkyl is a mono-ringed
group. Preferably a C.sub.3-C.sub.7 cycloalkyl group, particularly
preferred are cyclopentane, cyclohexane and cycloheptane groups,
e.g. cyclopentane or cyclohexane. In another embodiment, the
cycloalkyl is a multi-ringed group, e.g. adamantyl.
[0176] As used herein, the term "heterocyclyl" refers to
heteroaryl, heterocycloalkyl and heterocycloalkenyl groups. The
term "heteroaryl" refers to an aryl group as defined above wherein
at least one ring atom is a heteroatom. Suitable heteroatoms will
be apparent to those skilled in the art and include, for example,
sulphur, nitrogen, oxygen, phosphorus and silicon. Particularly
preferred is when the heteroatom is sulphur, nitrogen or
oxygen.
[0177] Monocyclic heteroaryl groups include for example, furan,
pyrrole, thiophene, imidazole, oxazole, thiazole,
1,3,4-thiadiazole, isothiazole, isoxazole, oxadiazole, oxazole,
1,2,3-oxadiazole, pyrazole, triazole, tetrazole, pyridine,
pyrazine, pyrimidine, pyridazines, triazine and tetrazine. Bicyclic
or polycyclic heteroaryl groups may include a monocyclic heteroaryl
group as defined herein, fused to one or more groups independently
selected from an aryl group, a cycloalkyl group, a cycloalkenyl
group and another monocyclic heteroaryl group. For example, the
heteroaryl group may be indole, benzimidazole, benzothiazole,
benzofuran, indoline, quinolone, isoquinoline, isoindole, indazole,
phenylpiperidine or benzothiene.
[0178] The terms "heterocycloalkyl" and "heterocycloalkenyl"
respectively refer to a cycloalkyl group or a cycloalkenyl group as
defined above, wherein at least one ring atom in the cycloalkyl or
cycloalkenyl group is a heteroatom. Again, suitable heteroatoms
will be apparent to those skilled in the art and include, for
example, sulphur, nitrogen, oxygen, phosphorus and silicon.
Particularly preferred is when the heteroatom is sulphur, nitrogen
or oxygen, e.g. aziridine, tetrahydrofuran, pyrrolidine, pyrroline,
piperidine, piperazine, thiazolidine, oxazolidine, morpholine,
thiane, thiazine, pyrazolidine, pyrazoline, imidazolidine or
imidazoline.
[0179] The term "alkoxy" refers to an O-alkyl group, wherein alkyl
is as defined above. Preferably, the alkoxy group is a C.sub.1-20
alkoxy group, more preferably a C.sub.1-15 alkoxy group, more
preferably still a C.sub.1-10 alkoxy group, more preferably still a
C.sub.1-8 alkoxy group, more preferably still a C.sub.1-6 alkoxy
group. Particularly preferred alkoxy groups include, for example,
methoxy, ethoxy, iso-propoxy, propoxy, butoxy, iso-butoxy, pentoxy
and hexyloxy.
[0180] Further preferred antimicrobial compounds for use in the
present invention are those capable of killing clinically latent
microorganisms. Methods for determining activity against clinically
latent bacteria include a determination, under conditions known to
those skilled in the art (such as those described in Nature
Reviews, Drug Discovery, 1, 895-910 (2002), the disclosures of
which are hereby incorporated by reference), of Minimum
Stationary-cidal Concentration ("MSC") or Minimum Dormicidal
Concentration ("MDC") for a test compound. A suitable compound
screening method against clinically latent microorganisms is
described in WO2000028074, the contents of which are incorporated
herein by reference as if the publication was specifically and
fully set forth herein.
[0181] Compounds for use according to the invention may be
administered as the raw material but the active ingredients are
preferably provided in the form of pharmaceutical compositions.
[0182] The active ingredients may be used either as separate
formulations or as a single combined formulation. When combined in
the same formulation it will be appreciated that the two compounds
must be stable and compatible with each other and the other
components of the formulation.
[0183] Formulations of the invention include those suitable for
oral, parenteral (including subcutaneous e.g. by injection or by
depot tablet, intradermal, intrathecal, intramuscular e.g. by depot
and intravenous), rectal and topical (including dermal, buccal and
sublingual) or in a form suitable for administration by inhalation
or insufflation administration. The most suitable route of
administration may depend upon the condition and disorder of the
patient. Preferably, the compositions of the invention are
formulated for oral or topical administration. In a preferred
embodiment, the composition is a cream or an ointment adapted for
nasal administration, in particular for delivery to the anterior
nares.
[0184] The formulations may conveniently be presented in unit
dosage form and may be prepared by any of the methods well known in
the art of pharmacy e.g. as described in "Remington: The Science
and Practice of Pharmacy", Lippincott Williams and Wilkins,
21.sup.st Edition, (2005). Suitable methods include the step of
bringing into association to active ingredients with a carrier
which constitutes one or more excipients. In general, formulations
are prepared by uniformly and intimately bringing into association
the active ingredients with liquid carriers or finely divided solid
carriers or both and then, if necessary, shaping the product into
the desired formulation. It will be appreciated that when the two
active ingredients are administered independently, each may be
administered by a different means.
[0185] When formulated with excipients, the active ingredients may
be present in a concentration from 0.1 to 99.5% (such as from 0.5
to 95%) by weight of the total mixture; conveniently from 30 to 95%
for tablets and capsules and 0.01 to 50% (such as from 3 to 50%)
for liquid preparations.
[0186] Formulations suitable for oral administration may be
presented as discrete units such as capsules, cachets or tablets
(e.g. chewable tablets in particular for paediatric
administration), each containing a predetermined amount of active
ingredient; as powder or granules; as a solution or suspension in
an aqueous liquid or non-aqueous liquid; or as an oil-in-water
liquid emulsion or water-in-oil liquid emulsion. The active
ingredients may also be presented a bolus, electuary or paste.
[0187] A tablet may be made by compression or moulding, optionally
with one or more excipients. Compressed tablets may be prepared by
compressing in a suitable machine the active ingredient in a
free-flowing form such as a powder or granules, optionally mixed
with other conventional excipients such as binding agents (e.g.
syrup, acacia, gelatin, sorbitol, tragacanth, mucilage of starch,
polyvinylpyrrolidone and/or hydroxymethyl cellulose), fillers (e.g.
lactose, sugar, microcrystalline cellulose, maize-starch, calcium
phosphate and/or sorbitol), lubricants (e.g. magnesium stearate,
stearic acid, talc, polyethylene glycol and/or silica),
disintegrants (e.g. potato starch, croscarmellose sodium and/or
sodium starch glycolate) and wetting agents (e.g. sodium lauryl
sulphate). Moulded tablets may be made by moulding in a suitable
machine a mixture of the powdered active ingredient with an inert
liquid diluent. The tablets may be optionally coated or scored and
may be formulated so as to provide controlled release (e.g.
delayed, sustained, or pulsed release, or a combination of
immediate release and controlled release) of the active
ingredients.
[0188] Alternatively, the active ingredients may be incorporated
into oral liquid preparations such as aqueous or oily suspensions,
solutions, emulsions, syrups or elixirs. Formulations containing
the active ingredients may also be presented as a dry product for
constitution with water or another suitable vehicle before use.
Such liquid preparations may contain conventional additives such as
suspending agents (e.g. sorbitol syrup, methyl cellulose,
glucose/sugar syrup, gelatin, hydroxymethyl cellulose,
carboxymethyl cellulose, aluminium stearate gel and/or hydrogenated
edible fats), emulsifying agents (e.g. lecithin, sorbitan
mono-oleate and/or acacia), non-aqueous vehicles (e.g. edible oils,
such as almond oil, fractionated coconut oil, oily esters,
propylene glycol and/or ethyl alcohol), and preservatives (e.g.
methyl or propyl p-hydroxybenzoates and/or sorbic acid).
[0189] Topical compositions, which are useful for treating
disorders of the skin or of membranes accessible by digitation
(such as membrane of the mouth, vagina, cervix, anus and rectum),
include creams, ointments, lotions, sprays, gels and sterile
aqueous solutions or suspensions. As such, topical compositions
include those in which the active ingredients are dissolved or
dispersed in a dermatological vehicle known in the art (e.g.
aqueous or non-aqueous gels, ointments, water-in-oil or
oil-in-water emulsions). Constituents of such vehicles may comprise
water, aqueous buffer solutions, non-aqueous solvents (such as
ethanol, isopropanol, benzyl alcohol, 2-(2-ethoxyethoxy)ethanol,
propylene glycol, propylene glycol monolaurate, glycofurol or
glycerol), oils (e.g. a mineral oil such as a liquid paraffin,
natural or synthetic triglycerides such as Miglyol.TM., or silicone
oils such as dimethicone). Depending, inter alia, upon the nature
of the formulation as well as its intended use and site of
application, the dermatological vehicle employed may contain one or
more components selected from the following list: a solubilising
agent or solvent (e.g. a .beta.-cyclodextrin, such as hydroxypropyl
.beta.-cyclodextrin, or an alcohol or polyol such as ethanol,
propylene glycol or glycerol); a thickening agent (e.g.
hydroxymethyl cellulose, hydroxypropyl cellulose, carboxymethyl
cellulose or carbomer); a gelling agent (e.g. a
polyoxyethylene-polyoxypropylene copolymer); a preservative (e.g.
benzyl alcohol, benzalkonium chloride, chlorhexidine, chlorbutol, a
benzoate, potassium sorbate or EDTA or salt thereof); and pH
buffering agent(s) (e.g. a mixture of dihydrogen phosphate and
hydrogen phosphate salts, or a mixture of citric acid and a
hydrogen phosphate salt). Topical formulations may also be
formulated as a transdermal patch.
[0190] Methods of producing topical pharmaceutical compositions
such as creams, ointments, lotions, sprays and sterile aqueous
solutions or suspensions are well known in the art. Suitable
methods of preparing topical pharmaceutical compositions are
described, e.g. in WO9510999, U.S. Pat. No. 6,974,585,
WO2006048747, as well as in documents cited in any of these
references.
[0191] Topical pharmaceutical compositions according to the present
invention may be used to treat a variety of skin or membrane
disorders, such as infections of the skin or membranes (e.g.
infections of nasal membranes, axilla, groin, perineum, rectum,
dermatitic skin, skin ulcers, and sites of insertion of medical
equipment such as i.v. needles, catheters and tracheostomy or
feeding tubes) with any of the bacteria, fungi described above,
(e.g. any of the Staphylococci, Streptococci, Mycobacteria or
Pseudomonas organisms mentioned hereinbefore, such as S. aureus
(e.g. Methicillin resistant S. aureus (MRSA))).
[0192] Particular bacterial conditions that may be treated by
topical pharmaceutical compositions of the present invention also
include the skin- and membrane-related conditions disclosed
hereinbefore, as well as: acne vulgaris; rosacea (including
erythematotelangiectatic rosacea, papulopustular rosacea, phymatous
rosacea and ocular rosacea); erysipelas; erythrasma; ecthyma;
ecthyma gangrenosum; impetigo; paronychia; cellulitis; folliculitis
(including hot tub folliculitis); furunculosis; carbunculosis;
staphylococcal scalded skin syndrome; surgical scarlet fever;
streptococcal peri-anal disease; streptococcal toxic shock
syndrome; pitted keratolysis; trichomycosis axillaris; pyoderma;
external canal ear infections; green nail syndrome; spirochetes;
necrotizing fasciitis; Mycobacterial skin infections (such as lupus
vulgaris, scrofuloderma, warty tuberculosis, tuberculides, erythema
nodosum, erythema induratum, cutaneous manifestations of
tuberculoid leprosy or lepromatous leprosy, erythema nodosum
leprosum, cutaneous M. kansasii, M. malmoense, M. szulgai, M.
simiae, M. gordonae, M. haemophilum, M. avium, M. intracellulare,
M. chelonae (including M. abscessus) or M. fortuitum infections,
swimming pool (or fish tank) granuloma, lymphadenitis and Buruli
ulcer (Bairnsdale ulcer, Searles' ulcer, Kakerifu ulcer or Toro
ulcer)); as well as infected eczma, burns, abrasions and skin
wounds.
[0193] Compositions for use according to the invention may be
presented in a pack or dispenser device which may contain one or
more unit dosage forms containing the active ingredients. The pack
may, e.g. comprise metal or plastic foil, such as a blister pack.
Where the compositions are intended for administration as two
separate compositions these may be presented in the form of a twin
pack.
[0194] Pharmaceutical compositions may also be prescribed to the
patient in "patient packs" containing the whole course of treatment
in a single package, usually a blister pack. Patient packs have an
advantage over traditional prescriptions, where a pharmacist
divides a patients' supply of a pharmaceutical from a bulk supply,
in that the patient always has access to the package insert
contained in the patient pack, normally missing in traditional
prescriptions. The inclusion of the package insert has been shown
to improve patient compliance with the physician's
instructions.
[0195] Suitable dosages and formulations for the administration of
colistin are described in the product label for Colomycin.RTM.
which can be found at
http://www.medicines.org/uk/emc/medicine/6301/SPC/Colomycin+Tablets/
[0196] Suitable dosages and formulations for the administration of
caffeic acid (hydroxycinnamic acid), thymol, aspirin, benzydamine
hydrochloride, diclofenac sodium, flurbiprofen, ibuprofen,
indomethacin, trifluoperazine hydrochloride, chlorprothixene
hydrochloride, triflupromazine hydrochloride, suloctidil,
thioridazine hydrochloride, dichlorophen, saccharin and piroxicam
may be obtained from conventional sources such as
www.medicine.org.uk,
http://www.accessdata.fda.gov/scripts/cder/drugsatfda/index,cfm,
www.rxlist.com and/or www.drugs.com. These sources disclose the
therapeutic, safe doses for each of these drugs. When used in
combination in accordance with the present invention, the dosage of
said drug may be decreased from that known.
[0197] The administration of the combination of the invention by
means of a single patient pack, or patients packs of each
composition, including a package insert directing the patient to
the correct use of the invention is a desirable feature of this
invention.
[0198] According to a further embodiment of the present invention
there is provided a patient pack comprising at least one active
ingredient of the combination according to the invention and an
information insert containing directions on the use of the
combination of the invention.
[0199] In another embodiment of the invention, there is provided a
double pack comprising in association for separate administration,
an antimicrobial agent, preferably having biological activity
against clinically latent microorganisms, and one or more of the
compounds disclosed herein preferably having biological activity
against clinically latent microorganisms.
[0200] According to a further embodiment of the present invention
there is provided a patient pack comprising at least one active
ingredient of the combinations according to the invention, i.e. at
least one of suloctidil or a pharmaceutically acceptable derivative
or prodrug thereof and a polymyxin selected from polymyxin E and
polymyxin B or a pharmaceutically acceptable derivative thereof,
and an information insert containing directions on the use of the
combination.
[0201] The amount of active ingredients required for use in
treatment will vary with the nature of the condition being treated
and the age and condition of the patient, and will ultimately be at
the discretion of the attendant physician or veterinarian. In
general however, doses employed for adult human treatment will
typically be in the range of 0.02 to 5000 mg per day, preferably 1
to 1500 mg per day. The desired dose may conveniently be presented
in a single dose or as divided doses administered at appropriate
intervals, e.g. as two, three, four or more sub-doses per day.
[0202] Biological Tests
[0203] Test procedures that may be employed to determine the
biological (e.g. bactericidal or antimicrobial) activity of the
active ingredients include those known to persons skilled in the
art for determining: [0204] (a) bactericidal activity against
clinically latent bacteria; and [0205] (b) antimicrobial activity
against log phase bacteria.
[0206] In relation to (a) above, methods for determining activity
against clinically latent bacteria include a determination, under
conditions known to those skilled in the art (such as those
described in Nature Reviews, Drug Discovery 1, 895-910 (2002), the
disclosures of which are hereby incorporated by reference), of
Minimum Stationary-cidal Concentration ("MSC") or Minimum
Dormicidal Concentration ("MDC") for a test compound.
[0207] By way of example, WO2000028074 describes a suitable method
of screening compounds to determine their ability to kill
clinically latent microorganisms. A typical method may include the
following steps: [0208] (1) growing a bacterial culture to
stationery phase; [0209] (2) treating the stationery phase culture
with one or more antimicrobial agents at a concentration and or
time sufficient to kill growing bacteria, thereby selecting a
phenotypically resistant sub-population; [0210] (3) incubating a
sample of the phenotypically resistant subpopulation with one or
more test compounds or agents; and [0211] (4) assessing any
antimicrobial effects against the phenotypically resistant
subpopulation.
[0212] According to this method, the phenotypically resistant
sub-population may be seen as representative of clinically latent
bacteria which remain metabolically active in vivo and which can
result in relapse or onset of disease.
[0213] In relation to (b) above, methods for determining activity
against log phase bacteria include a determination, under standard
conditions (i.e. conditions known to those skilled in the art, such
as those described in WO 2005014585, the disclosures of which
document are hereby incorporated by reference), of Minimum
Inhibitory Concentration ("MIC") or Minimum Bactericidal
Concentration ("MBC") for a test compound. Specific examples of
such methods are described below.
EXAMPLES
[0214] The chequerboard method and Time kill experiments are
described below and in Antimicrob Chemo (2013) 68, 374-384.
Example 1: Time Kill Experiments
[0215] (a) Thymol (HT013015) and Colistin Against NDM-1 Klebsiella
pneumoniae
[0216] FIG. 1 is the time kill curve showing combination of
colistin and HT013015 against NDM-1 K.pneumoniae.
[0217] (b): Trifluoperazine Hydrochloride (HT0120566) and Colistin
Against NDM-1 Klebsiella pneumoniae
[0218] FIG. 2 is the time kill curve showing combination of
colistin and HT0120566 against NDM-1 Klebsiella pneumoniae.
[0219] (c): Chlorprothixene Hydrochloride (HT0120707) and Colistin
Against NDM-1 E. coli
[0220] FIG. 3 is the time kill curve showing combination of
colistin and HT0120707 against NDM-1 E. coli.
[0221] (d): Triflupromazine Hydrochloride (HT0120700) and Colistin
Against NDM-1 E. coli
[0222] FIG. 4 is the time kill curve showing combination of
colistin and HT0120700 against NDM-1 E. coli.
[0223] (e): Suloctidil (HT0120093) and Colistin Against NDM-1 E.
coli
[0224] FIG. 5 is the time kill curve showing combination of
colistin and HT0120093 against NDM-1 E. coli.
[0225] (f): Thioridazine Hydrochloride (HT0120553) and Colistin
Against NDM-1 Klebsiella pneumoniae
[0226] FIG. 6 is the time kill curve showing combination of
colistin and HT0120553 against NDM-1 Klebsiella pneumoniae.
[0227] (g): Dichlorophen (HT0121567) and Colistin Against NDM-1 K.
pneumoniae
[0228] FIG. 7 is the time kill curve showing combination of
colistin and HT0121567 against NDM-1 K.pneumoniae.
[0229] (h): Aspirin (HT0121219) and Colistin Against NDM-1
Klebsiella pneumoniae
[0230] FIG. 8 is the time kill curve showing combination of
colistin and HT0121219 against NDM-1 K.pneumoniae
[0231] (i): Indomethacin (HT0120451) and Colistin Against NDM-1 K.
pneumoniae
[0232] FIGS. 9 and 10 are the time kill curves showing combination
of colistin and HT0120451 against NDM-1 K.pneumoniae.
[0233] (j): Piroxicam (HT0120492) and Colistin Against NDM-1 E.
coli
[0234] FIG. 11 is the time kill curve showing combination of
colistin and HT0120492 against NDM-1 E. coli.
[0235] (k): Benzydamine Hydrochloride (HT0120969) and Colistin
Against NDM-1 E. coli
[0236] FIG. 12 is the time kill curve showing combination of
colistin and HT0120969 against NDM-1 E. coli.
[0237] (I): Ibuprofen (HT0120448) and Colistin Against NDM-1 K.
pneumoniae
[0238] FIG. 13 is the time kill curve showing combination of
colistin and HT0120448 against NDM-1 Klebsiella pneumoniae.
[0239] (m): Diclofenac Sodium (HT0120365) and Colistin Against
NDM-1 E. coli
[0240] FIG. 14 is the time kill curve showing combination of
colistin and HT0120365 against NDM-1 E. coli.
[0241] (n): Flurbiprofen (HT0120417) and Colistin Against NDM-1 E.
coli
[0242] FIG. 15 is the time kill curve showing combination of
colistin and HT0120417 against NDM-1 E. coli
[0243] (o): Caffeic Acid (HT013001) and Colistin Against NDM-1 E.
coli
[0244] FIG. 16 is the time kill curve showing combination of
colistin and HT013001 against NDM-1 E. coli
[0245] (p): Saccharin (HT0120098) and Colistin Against NDM-1 E.
coli
[0246] FIG. 17 is the time kill curve showing combination of
colistin and HT0120098 against NDM-1 E. coli.
Example 2: Chequerboard Method
[0247] (a) In Vitro Synergy Effect of Colistin and Each of HT013015
(Thymol), HT0121219 (Aspirin), HT0120448 (Ibuprofen), HT0120451
(Indomethacin), HT0120566 (Trifluoperazine Hydrochloride), and
HT0121567 (Dichlorophen) Against Log Phase NDM-1 Klebsiella
pneumoniae Using the Chequerboard Method
[0248] Growth of Bacteria
[0249] Log phase growth of NDM-1 Klebsiella pneumonia was carried
out as described in the art.
[0250] The effects of each combination of the present invention
were examined by calculating the fractional inhibitory
concentration index (FICI) of each combination, as follows: (MIC of
drug A, tested in combination)/(MIC of drug A, tested alone)+(MIC
of drug B, tested in combination)/(MIC of drug B, tested alone).
The interaction of the combination was defined as showing synergy
if the FICI was .ltoreq.3.5, no interaction if the FICI was >0.5
but <4.0 and antagonism if the FICI was >4.0. The results are
depicted in FIGS. 35-40.
[0251] (b) In Vitro Synergy Effect of Colistin and Each of
HT0120969 (Benzydamine Hydrochloride), HT0120365 (Diclofenac
Sodium), HT0120417 (Flurbiprofen), HT0120707 (Chlorprothixene
Hydrochloride), HT0120700 (Triflupromazine Hydrochloride),
HT0120093 (Suloctidil), HT0120492 (Piroxicam) and HT0120098
(Saccharin) Against Log Phase NDM-1 E. coli Using the Chequerboard
Method
[0252] Growth of Bacteria
[0253] Log phase growth of NDM-1 E. coli was carried out as
described in the art.
[0254] The effects of each combination of the present invention
were examined by calculating the fractional inhibitory
concentration index (FICI) of each combination, as follows: (MIC of
drug A, tested in combination)/(MIC of drug A, tested alone)+(MIC
of drug B, tested in combination)/(MIC of drug B, tested
alone).
[0255] The interaction of the combination was defined as showing
synergy if the FICI was .ltoreq.3.5, no interaction if the FICI was
>0.5 but <4.0 and antagonism if the FICI was >4.0. The
results are depicted in FIGS. 41-49.
Example 3
[0256] In Vitro Synergy Effect of Suloctidil in Combination with
Colistin Against Log Phase NDM-1 Klebsiella pneumoniae
[0257] The synergistic effect of suloctidil in combination with
colistin was tested against log phase NDM-1 Klebsiella pneumoniae
using time-kill methods over a period of 24 hours.
[0258] Materials and Methods [0259] Bacterial strain used: NCTC
13443 strain of NDM-1 Klebsiella pneumoniae [0260] Growth of
bacteria: Log phase growth of the bacteria was carried out
according to methods known in the art.
[0261] Compounds and Preparation:
[0262] (i) Suloctidil was obtained from a commercial source and
dissolved in DMSO to make a stock concentration of 10 mg/ml.
[0263] (ii) Colistin was obtained from a commercial source at a
concentration of 20 mg/ml.
[0264] Both suloctidil and colistin were then added to 96 well
plates either alone or in the combinations shown below in Table
1.
TABLE-US-00001 TABLE 1 Agent (Concentration) Number/Letter
Combination Combination Colistin (32 .mu.g/ml) 1 1&A 1&C
Colistin (16 .mu.g/ml) 2 2&A 2&C Colistin (8 .mu.g/ml) 3
3&A 3&C Colistin (4 .mu.g/ml) 4 4&A 4&C Suloctidil
(32 .mu.g/ml) A 1&B 1&D Suloctidil (16 .mu.g/ml) B 2&B
2&D Suloctidil (8 .mu.g/ml) C 3&B 3&D Suloctidil (4
ug/ml) D 4&B 4&D
[0265] The overnight culture was diluted with nutrient broth
(Oxoid) to 10.sup.7 CFU/ml and 280 .mu.l and 290 .mu.l of the
culture was added to each combination well and single agent well,
respectively, to make a final concentration of 300 .mu.l.
Incubation of the compounds with the bacterial suspension was
carried out for 24 hours. At 0, 2, 4, 7 and 25 hours, CFU counts
were performed to measure the kill effects of the drug
combination.
[0266] Results
[0267] The time-kill curves are shown in FIGS. 18 to 27, where
suloctidil is referenced as "93".
[0268] Summary and Conclusion [0269] 1. It can be seen from FIGS.
18 to 27 that suloctidil (i.e. "93") had no antimicrobial effect
against log phase NDM-1 K.pneumoniae when used alone at
concentrations of 32, 16, 8 and 4 .mu.g/ml. [0270] 2. It can be
seen from FIGS. 20, 21, 24, 25 and 27 that colistin also had no
antimicrobial effect against log phase NDM-1 K.pneumoniae when used
alone at concentrations of 8 and 4 .mu.g/ml. [0271] 3. FIGS. 18,
19, 22, 23 and 26 then demonstrate that colistin had an
antimicrobial effect against log phase NDM-1 K.pneumoniae at
concentrations of 32 and 16 .mu.g/ml, but that this effect was not
long term. [0272] 4. FIG. 19 for example shows that colistin (16
.mu.g/ml) caused complete kill after 7 hours and bacteria re-growth
after 25 hours. [0273] 5. In each of FIGS. 18 to 27, it can,
however, be seen that there was a significant synergistic effect
against log phase NDM-1 K.pneumoniae when suloctidil and colistin
were used in combination. This synergistic effect resulted in both
a faster and longer term kill of the bacteria.
Example 4: In Vitro Synergy Effect of Suloctidil in Combination
with Polymyxin E (Colistin) Against Log Phase NDM-1 E. coli
[0274] The synergistic effect of suloctidil in combination with
colistin was tested against log phase NDM-1 E. coli using time-kill
methods over a period of 24 hours.
[0275] Materials and Methods [0276] Bacterial strain used: BAA2469
strain of NDM-1 E. coli [0277] Growth of bacteria: Log phase growth
of bacteria was carried out according to methods known in the
art.
[0278] Compounds and Preparation:
[0279] (i) Suloctidil was obtained from a commercial source and
dissolved in DMSO to make a stock concentration of 10 mg/ml.
[0280] (ii) Colistin was obtained from a commercial source at a
concentration of 20 mg/ml. Both suloctidil and colistin were then
added to 96 well plates either alone or in the combinations shown
below in Table 2.
TABLE-US-00002 TABLE 2 Agent (Concentration) Number/Letter
Combination Combination Colistin (8 .mu.g/ml) 1 1&A 1&C
Colistin (4 .mu.g/ml) 2 2&A 2&C Colistin (2 .mu.g/ml) 3
3&A 3&C Colistin (1 .mu.g/ml) 4 4&A 4&C Suloctidil
(32 .mu.g/ml) A 1&B 1&D Suloctidil (16 .mu.g/ml) B 2&B
2&D Suloctidil (8 .mu.g/ml) C 3&B 3&D Suloctidil (4
ug/ml) D 4&B 4&D
[0281] The overnight culture was diluted with nutrient broth
(Oxoid) to 10.sup.7 CFU/ml and 280 .mu.l and 290 .mu.l of the
culture was added to each combination well and single agent well,
respectively, to make a final concentration of 300 .mu.l.
Incubation of the compounds with the bacterial suspension was
carried out for 24 hours. At 0, 2, 4, 7 and 25 hours, CFU counts
were performed to measure the kill effects of the drug
combination.
[0282] Results
[0283] The time-kill curves are shown in FIGS. 28 to 34, where
suloctidil is referenced as "93".
SUMMARY AND CONCLUSION
[0284] 1. It can be seen from FIGS. 28 to 34 that suloctidil (i.e.
"93") had no antimicrobial effect against log phase NDM-1 E. coli
when used alone at concentrations of 16, 8 and 4 .mu.g/ml. [0285]
2. It can be seen from FIGS. 28 to 34 that colistin also had no
antimicrobial effect against log phase NDM-1 E. coli when used
alone at concentrations of 8, 4, 2 and 1 .mu.g/ml. [0286] 3. In
each of FIGS. 28 to 34, it can, however, be seen that there was a
significant synergistic effect against log phase NDM-1 E. coli when
suloctidil and colistin were used in combination. This synergistic
effect resulted in a faster time-kill of the bacteria.
* * * * *
References