WO2002020478A1 - Derives d'acide acetique n-benzylindole-3 pouvant etre utilises dans le traitement de cancers pharmacoresistants - Google Patents

Derives d'acide acetique n-benzylindole-3 pouvant etre utilises dans le traitement de cancers pharmacoresistants Download PDF

Info

Publication number
WO2002020478A1
WO2002020478A1 PCT/IE2001/000109 IE0100109W WO0220478A1 WO 2002020478 A1 WO2002020478 A1 WO 2002020478A1 IE 0100109 W IE0100109 W IE 0100109W WO 0220478 A1 WO0220478 A1 WO 0220478A1
Authority
WO
WIPO (PCT)
Prior art keywords
compound
mrp
cancer
drug
methoxy
Prior art date
Application number
PCT/IE2001/000109
Other languages
English (en)
Inventor
Michael Clynes Martin
Robert Andrew O'connor
Samantha Mary Touhey
Anita Rose Maguire
Stephen James Plunkett
Original Assignee
Dublin City University
National University Or Ireland, Cork
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Dublin City University, National University Or Ireland, Cork filed Critical Dublin City University
Priority to AU2001282410A priority Critical patent/AU2001282410A1/en
Publication of WO2002020478A1 publication Critical patent/WO2002020478A1/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
    • C07F9/553—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having one nitrogen atom as the only ring hetero atom
    • C07F9/572—Five-membered rings
    • C07F9/5728—Five-membered rings condensed with carbocyclic rings or carbocyclic ring systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • A61K31/403—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil condensed with carbocyclic rings, e.g. carbazole
    • A61K31/404—Indoles, e.g. pindolol
    • A61K31/405—Indole-alkanecarboxylic acids; Derivatives thereof, e.g. tryptophan, indomethacin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D209/00—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom
    • C07D209/02—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom condensed with one carbocyclic ring
    • C07D209/04—Indoles; Hydrogenated indoles
    • C07D209/10—Indoles; Hydrogenated indoles with substituted hydrocarbon radicals attached to carbon atoms of the hetero ring
    • C07D209/18—Radicals substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
    • C07D209/22—Radicals substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals with an aralkyl radical attached to the ring nitrogen atom
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • the invention relates to N-benzylrndole-3-acetic acid derivatives for use in the prophylaxis and/or treatment of drug resistant cancer.
  • cytotoxic agents have been developed that can improve the treatment outcome of many cancers, including solid tumours.
  • the natural product isolates and their derivatives - including anthracyclines, vinca alkaloids, epipodophyllotoxins, and taxanes - have proved to be effective in inducing remissions and cures in many malignancies .
  • Resistance to drugs may be due to one or more general mechanisms including, changes in drug distribution in the body (for example modifications of blood flow), changes in cellular permeability to the drug in question, modifications in target mechanisms (e.g. modifications of the topoisomerase enzyme), adaptive changes in the metabolism of the drug, changes in the downstream effectors of drug action (e.g. inhibition of apoptosis mechanism in cancer treatment), sequestration of drug into areas of the cell where it cannot affect the cellular target or active efflux of the drug from the target cell 5 .
  • modifications in target mechanisms e.g. modifications of the topoisomerase enzyme
  • adaptive changes in the metabolism of the drug e.g. inhibition of apoptosis mechanism in cancer treatment
  • sequestration of drug into areas of the cell where it cannot affect the cellular target or active efflux of the drug from the target cell 5 e.g. inhibition of apoptosis mechanism in cancer treatment
  • P-glycoprotein also known as PI 70 6 ' 7,8 .
  • This pump utilises the energy of ATP to actively efflux a variety of agents including a broad spectrum of cancer chemotherapeutic drugs 9 .
  • the net effect of this efflux mechanism is to reduce the time a cell is exposed to the agent and to reduce its maximum intracellular concentration. Since most cytotoxic cancer therapies are given at or near the maximum tolerable dose, and have little selectivity for cancer cells, the dose given cannot be increased to compensate without becoming toxic for the patient.
  • P-gp can pump a wide variety of structurally dissimilar agents including many of the more effective, first-line anti-cancer drugs such as doxorubicin and vinca alkaloids 10 . If cells already express a high level of P-gp activity they will be intrinsically resistant to the cytotoxic effects of these agents. Likewise, if P-gp expression is induced, or a P-gp-overexpressing subpopulation is selected, in a tumour by initial cytotoxic exposure, all of the other substrate agents will be less effective if given subsequently 10 . Because P-gp was the earliest multidrug resistance (MDR) mechanism to be investigated clinically, a significant amount of information has amassed on the clinical impact of P-gp inhibition in different cancers and with different combinations of drugs.
  • MDR multidrug resistance
  • MRP multidrug resistance protein
  • MRP-1 the most studied member of the family, has similar (but not identical) substrate specificities to P-gp but the two proteins only share a 30 % similarity in protein sequence 12 . It is accepted in the literature that
  • MRP substrates include the anthracyclines (e.g. doxorubicin or epirubicin), epipodophyllotoxins (e.g.etoposide), vinca alkaloids (especially vincristine) methotrexate and taxanes (especially taxol) 15 ' 16 .
  • anthracyclines e.g. doxorubicin or epirubicin
  • epipodophyllotoxins e.g.etoposide
  • vinca alkaloids especially vincristine
  • methotrexate especially taxanes 15 ' 16 .
  • MRP-1 is over-expressed in a number of multidrug-resistant cell lines, but, more interestingly, it now appears that it may be over-expressed, ahead of the over- expression of P-170, in a variety of human cancers, including breast carcinoma 17,18 .
  • MRP is also present in many normal cells but over-expression in tumour cells appears to be associated with poor prognostic outcome in several forms of cancer and chemotherapeutic treatment 19 .
  • MRP expression in drug resistance remains to be fully explored, although evidence suggests a correlation with prognosis or impact of chemotherapy in neuroblastoma 20 , retinoblastoma 21 , primary breast carcinoma 22 , relapsed acute leukemia 23 , acute myeloid leukemia 24 , malignant melanoma 25 , and non-small cell lung cancer 26 .
  • MRP-1 Inhibition of MRP-1 has been suggested as a mechanism to improve the cancer therapy with MRP-1 substrate drugs. To date no clinical trials of specific MRP-1 inhibitors have been initiated but several agents have been suggested as viable, specific inhibitors of MRP for clinical use 27 .
  • NSAIDs are currently used to treat a variety of inflammatory conditions where their therapeutic activity is associated with the ability to inhibit the enzyme cyclooxygenase (COX) in the body.
  • COX cyclooxygenase
  • Two forms of cyclooxygenase have been described, a constitutively expressed variant COX-1, which is largely responsible for cytoprotective mechanisms, such as those operating in the stomach lining, and COX-2, which is an inducible form, primarily responsible for a variety of inflammatory reactions 1,32 .
  • COX-2 activity has also been associated with progression of certain forms of colon cancers and specific inhibitors of COX-2 have received approval for use in preventing the emergence of cancer in patients genetically predisposed to develop colon cancer as young adults (Familial Adenomatous Polyposis) 33 ' 34 ' 35 .
  • the NSAIDs currently in common clinical use generally have little specificity and inhibit both forms of COX. This means that prolonged use, which is often required with chronic immunological conditions such as arthritis, is commonly associated with negative effects such as gastrointestinal intolerance, gastro-intestinal ulceration and reduced renal blood flow (all associated with COX-1 inhibition). Agents such as indomethacin are also quite toxic directly and this limits the maximum dose which can be given acutely 36 .
  • Indomethacin in common with most other NSAIDs, has been shown to inhibit both COX-1 and COX-2 isoforms; the gastrotoxicity associated with this class of compounds is believed to be due to COX-1 inhibition and accordingly extensive searching for selective COX-2 inhibitors has been undertaken in recent years 39,4 °. Indeed the indomethacin derivative in which the benzoyl group is replaced by a
  • 4-bromobenzyl group has been reported as a highly selective COX-2 inhibitor 4I .
  • some NSAIDs also inhibit other enzymes, including phospholipase A 2 (PLA 2 ).
  • PPA 2 phospholipase A 2
  • Kaplan and coworkers found that indomethacin inhibits PLA 2 from rabbit polymorphonuclear leukocytes, ⁇ while more recent reports have studied the inhibitory abilities of other NSAIDs 43 .
  • MRP- inhibitory NSAIDs represent the most likely agents to provide therapeutically useful MRP inhibition.
  • MRP-inhibitory NSAIDS particularly indomethacin
  • This invention is directed towards providing inhibitors of MRP which provide for a similar level of MRP inhibition while having less toxicity and associated side effects than known MRP inhibitors.
  • X is H, Cl, Br, F with the provisos that:
  • the C ⁇ -C 6 alkyl are substituted with one or more of the same or different of:
  • the compound of Formula I has a carboxylic acid or a phosphonic acid functional group.
  • the pharmacologically acceptable salt of a compound of Formula I is an alkali metal salt selected from the group comprising sodium, potassium and lithium or an alkali earth metal salt selected from the group comprising magnesium and calcium.
  • Compounds of formula III have been found in particular to increase the effect of anti-cancer drugs such as adriamycin and taxol. Because the compounds have low toxicity and low side effects the compounds have major therapeutic potential in the prophylaxis and/ or treatment of drug resistant cancer.
  • One embodiment of the invention provides use of a compound of formula I, II or III for the prophylaxis and/ or treatment of multiple drug resistance related disease.
  • Another embodiment of the invention provides use of a compound of Formula I, II or III for the prophylaxis and/ or treatment of leishmaniasis.
  • One embodiment of the invention provides for the use of a compound of formula I, II or III with an identifiable marker, preferably the identifiable marker is a radiolabelled marker.
  • a further embodiment of the invention provides for the use of compounds of formula I, II or III for identifying tumours which are mutidrug resistant.
  • the invention also provides a pharmaceutical composition
  • a pharmaceutical composition comprising a compound of formula I, II or III as hereinbefore described.
  • the C r C D alkyl are substituted with one or more of the same or different of hydroxy, carboxy, phosphonic groups.
  • the compound of formula I has a carboxylic acid or a phosphonic acid functional group.
  • the compound of formula I comprises a pharmacologically acceptable salt preferably an alkali metal salt selected from the group comprising sodium, potassium and lithium or an alkali earth metal salt selected from the group comprising magnesium and calcium.
  • the MRP substrate drug is a MRP-1 substrate drug.
  • the substrate drug is an anti-cancer drug selected from the group comprising an anthracycline, a vinca alkaloid, an epipodophyllotoxin or a taxane.
  • the anti-cancer drug is selected from any one or more of anthracycline, adriamycin, daunorubicin, epirubicin, vinca alkaloid, vincristine, epipodophyllotoxin etoposide (VP-16), teniposide or taxol.
  • the pharmaceutical composition of the invention may be in the form of an emulsion, liposome, patch, powder and/or complex.
  • the composition comprises an ingestable carrier in which case the composition is in the form of a tablet, capsule, syrup or powder.
  • the pharamceutical composition is in combination with an agent which is a substrate for multidrug resistance related protein for simultaneous, sequential or separate use.
  • the pharmaceutical composition comprises an adjuvant such as an anti-emetic, an anti-inflammatory agent or a cancer chemotherapy medicament.
  • an adjuvant such as an anti-emetic, an anti-inflammatory agent or a cancer chemotherapy medicament.
  • the pharmaceutical composition of the invention is in a form for oral, intravenous, intramuscular, intraperitoneal, intradermal, intravesicular and/or rectal administration.
  • the invention provides a compound of formula I or pharmacologically acceptable salts thereof and an identificable marker, preferably the identifiable marker is a radiolabelled marker. These compounds are potentially therapeutically valuable in identifying whether tumours are mutidrug resistant or not.
  • the invention also provides a compound of the formula
  • the invention further provides a compound of the formula
  • X is F, Cl, Me or MeS.
  • the invention also provides a compound selected from
  • the invention further provides a compound selected from
  • the invention further provides a compound selected from
  • the invention also provides a pharmaceutical composition comprising a compound of the invention.
  • the composition comprises a compound of the invention and an MRP substrate drug.
  • the invention further provides use of a compound of formula I, II or III in the preparation of a medicament for the prophylaxis and/ or treatment of drug resistant cancer, multiple drug resistance related disease or leishmaniasis.
  • the compound may be present in the medicament at an amount suitable to produce a concentration in the bloodstream of from 20ng/ml to lOO ⁇ g/ml most preferably from l ⁇ g /ml to 20 ⁇ g/ml.
  • the compounds of the invention are likely to be less toxic than NSAID-based inhibitors and can also have less side effects on other enzymes including COX-1 or have specific effects on potentially damaging enzymes such as COX-2. This is likely to make these compounds more useful as they are likely to have better specificity and less toxicity and side effects.
  • N-Benzylindole-3-acetic acid derivatives which are active as inhibitors of MRP while having less direct toxicity, less COX -1 inhibitory activity and, in some cases, having useful activities such as COX-2 inhibition, in comparison to other known MRP inhibitors.
  • a number of these compounds are known per se, however none have previously been described as having MRP inhibitory activity 48 ' 60 ' 61 .
  • MRP-1 substrate drugs appear to be more useful in combination with MRP-1 substrate drugs in the treatment of, for example, drug resistant cancer or tumours likely to develop drug resistant cancers because they can be given at higher concentration (producing better inhibition of MRP-1) while being less toxic and having less side effects than other known MRP inhibitors. These compounds have been found to potentiate the toxicity of MRP substrate anti-cancer drugs.
  • the compounds may also be more beneficial in the treatment of diseases which are dependent on MRP-1 activity or where MRP-1 activity reduces the effectiveness of existing therapies.
  • MRP inhibitors of the present invention may be used with radiolabelled markers, and the like, attached, to identify whether tumours are multidrug resistant or not allowing the treatment regime to be altered accordingly.
  • N-benzyl derivatives compounds 5, 13, 15-17, 19 and 20 were found to be inactive in inhibiting MRP even though they have similar chemical structures to the active compounds of examples 1 to 4.
  • toxicity assays were performed which compared the effect of the drug adriamycin (doxorubicin) in the absence and presence of an inhibitor.
  • test compounds were prepared as described previously.
  • the DLKP ceU line used in these examples was derived from a lung cancer patient and developed by researchers in the National CeU and Tissue Culture Centre, Dublin City University.
  • the CORL23 (R) lung ceU line was a gift of
  • Indomethacin and aU other named chemicals used in this example were supplied by the Sigma Chemical Company Ltd. Fancy Road, Dorset, BH12 4QH,
  • CeUs were trypsinised from the flask in the exponential phase of growth. CeU suspensions containing lxlO 4 ceUs/ml were prepared in ceU culture medium. Volumes of lOO ⁇ l of this ceU suspension were added into 96-weU plates using a multichannel pipette. Plates were agitated gently in order to ensure even dispersion of ceUs over a given weU. CeUs were then incubated overnight at 37°C in an atmosphere containing 5% CO 2 .
  • Cytotoxic drug dUutions and test compound dUutions were prepared at 4X their final concentration in media. Nolumes of 50 ⁇ L of the drug dUution and 50 ⁇ L of the test compound dUution were then added to each relevant weU so that a total final volume of 200 ⁇ L was present in each weU.
  • AU potential toxicity-enhancing agents were dissolved in DMSO, ethanol or media. Stock solutions were prepared at approximately 15mg/10ml media, filter sterUised with a 0.22 ⁇ m filter and then used to prepare aU subsequent dUutions. Solvent control experiments showed that no toxicity enhancement effects were due to the presence of DMSO or ethanol.
  • CeUs were incubated for a further 6 days at 37°C in an atmosphere containing 5% C0 2 . At this point the control weUs would have reached approximately 80-
  • CeU number was assessed using the acid phosphatase assay. Each weU on the plate was washed with lOO ⁇ l PBS. This was then removed and lOO ⁇ l of freshly prepared phosphatase substrate (lOmM / nitrophenol phosphate in 0.1M sodium, 0.1% triton X-100, pH 5.5) was added to each weU.
  • the plates were then incubated in the dark at 37°C for 2 hours and the enzymatic reaction was stopped by the addition of 50 ⁇ l of IN ⁇ aOH.
  • the plate was read in a dual beam plate reader at 405nm with a reference wavelength of
  • % ceU survival in the drug-free control is 100%.
  • AU results are the ceU survival means ⁇ the standard deviation (SD) of that mean from a minimum of three experiments.
  • the derivative concentrations used in aU cases are non-toxic, i.e. there is no expectation of significantly increasing ceU death by combining the two drugs.
  • Indomethacin a known potentiator of toxicity in MRP-1-expressing ceU lines, is used as a positive control.
  • the Combination Index values quoted are a statistical measure of the toxic synergism (or lack of) between the compound and adriamycin. The smaUer the value the greater the synergism evident. Values of 1 or greater indicate only additive toxicity or antagonism of the toxic effect of adriamycin.
  • Table 1.11 Ulustrates that a synergistic increase in the toxicity of taxol was evident in the MRP-1 resistant ceU line COR L23 R when this anti cancer drug was combined with Compound 27.
  • Table 1.12 Ulustrates that this effect was not evident when the structuraUy simUar agent compound 28 was combined with taxol.
  • Table l.ll
  • MRP-1 protein MRP-1 protein.
  • IOVs Inside Out Vesicles
  • MRP-1 substrates such as radiolabeUed Leukotriene C
  • the activity of the pump can be measured by quantifying the total radioactivity of the vesicles over time.
  • Agents which inhibit the action of MRP-1 reduce the amount of radiation accumulated into the vesicles.
  • test compounds were prepared as described previously.
  • the PMSF was added to the buffer immediately before use. CeUs were lysed by gentle agitation at 4 °C for 1.5 hours. The ceU lysate was then centrifuged at 28,000 rpm. (100,000g) for 35 minutes at 4° C with a Beckman SW28 rotar in a Beckman XL-80 ultracentrifuge. The resulting peUets were then resuspended in 10ml of hypotonic buffer and then homogenised for 15 minutes at 4° C with a Braun Potter S886 homogeniser.
  • the homogenised ceU extract was dUuted to a final volume of 20ml with incubation buffer which contained the foUowing
  • the crude membrane fraction was layered over 38%> sucrose/ lOmM TRIS-HCl pH 7.4, (38g sucrose in 100ml lOmM TRIS HCl pH 7.4) and centrifuged at 28,000 r.p.m (100,000g) for 35 minutes at 4°C with a SW28 rotor.
  • peUets were then resuspended in 0.2ml incubation buffer. Vesicles were formed by passing resuspended peUets through a 27-gauge needle 20 times using a lml syringe.
  • a protein assay was then performed and the IOV preparation was then dUuted to a concentration of 5mg protein /ml with incubation buffer. Volumes of 50 ⁇ L IOVs were then frozen at -80° C.
  • Protein levels were determined using the Bio-Rad protein assay kit (Bio-Rad, 5000006) as foUows.
  • BSA bovine serum albumin
  • the ATP/creatine phosphate/MgCl 2 /10mM TRIS-HCl (pH 7.4) solution was prepared as foUows
  • thermomixer was placed in the thermomixer and the foUowing added sequentiaUy: 60 ⁇ L incubation buffer, 30 ⁇ L ATP, 5 ⁇ L creatine kinase, 5ul [ 3 H]- LTC 4 (DuPont NEN, NET-1018, 0.01 mCi/ml) and 10 ⁇ L IOVs. After every sequential addition the thermomixer was adjusted to half speed mixing to aUow agitation of the various components of the mixture.
  • the compound of interest was dissolved in incubation buffer at the desired concentration. 5 ⁇ L of this was added to an eppendorf in the thermomixer. 55 ⁇ L of incubation buffer was then added. The standard volumes of ATP, AMP, creatine kinase, LTC 4 and IOV were then added to a total final volume of HO ⁇ L.
  • Relative ATP-dependent rates shown are expressed as a percentage of untreated control, taken as 100%, by subtracting the rate in the presence of AMP, which was used as the blank. Data given are from a minimum of three assay repeats.
  • test compounds were prepared as described previously.
  • DLKP ceUs were seeded into 75cm 2 flasks (Costar, 3375) at 0.5x10° ceUs per flask. CeUs were incubated for 48 hours, after which time medium was removed and fresh medium containing adriamycin (lO ⁇ m), indomethacin or test compound (27.95 ⁇ m), or combination of both adriamycin and indomethacin/compound of interest, was added. Flasks were incubated at 37°C for a period of two hours.
  • the media was removed from aU flasks and replaced with fresh media, or media containing indomethacin/test compound or adriamycin, as the experiment required.
  • the flasks were returned to the 37°C incubator. At relevant time points the media was removed from the flasks and the flasks were washed twice with PBS. CeUs were then trypsinised and counted. PeUets were then washed with PBS and frozen at -20°C. When required for HPLC analysis, the frozen peUets were thawed, resuspended in lOO ⁇ l UHP water and added to glass tubes (test samples).
  • Untreated ceUs were resuspended in 800 ⁇ l UHP and lOO ⁇ l of this were placed in to 8 glass tubes. These were the adriamycin control tubes and were labeUed as foUows: 50 ⁇ g/ml, lO ⁇ g/ml, 5 ⁇ g/ml, 2 ⁇ g/ml, l ⁇ g/ml, 0.5 ⁇ g/ml, 0.25 ⁇ g/ml, O ⁇ g/ml adriamycin.
  • the HPLC mobUe phase was prepared as foUows: 64ml of 0.1M phosphoric acid (Sigma, P6560) was added to 488 UHP water. The pH was then adjusted to 2.3 with IN potassium hydroxide (Sigma P6310). A volume of 248 ml acetonitrile was added finally and the completed mobUe phase aUowed to degas at 4°C overnight.
  • the samples for analysis were automaticaUy injected in to the HPLC system (Beckman System Gold 507 autosampler, 125 pump and 166 detector). MobUe phase flow rate was set at 0.5ml per minute with a total run time of 16 minutes.
  • the column used for HPLC analysis of adriamycin in DLKP was a C 18 reversed phase Prodigy 5 ⁇ m particle size ODS-3 column (Phenomenex, U.K.). Absorbance was monitored at 253nm.
  • a standard curve of adriamycin peak area /daunorubicin internal standard versus adriamycin concentration was used to quantify the levels of adriamycin present in the samples. Results were finally reported as the content of adriamycin per miUion ceUs.
  • T5 Time point 5 hours after initial 2 hour loading period
  • (+) Indomethacin/analogue Flasks re-fed with either indomethacin or test compound (28 um) after initial 2 hour loading period.
  • (-) Indomethacin/test compound Flasks re-fed with media only after initial 2 hour loading period.
  • Indomethacin has inherent toxicity to ceUs and this may limit any future application to the field of MRP -resistance circumvention. Experiments were undertaken to examine the toxicity of these agents by measuring the highest concentration of agent which did not produce a toxic effect in the ceUs (highest non-toxic concentration).
  • test compounds were prepared as described previously.
  • CeUs in the exponential phase of growth were harvested by trypsinisation.
  • CeU suspensions containing lxlO 4 ceUs/ml were prepared in ceU culture medium. Volumes of lOO ⁇ l of these ceU suspensions were added in to 96 weU plates
  • Test compound dUutions were prepared at 2X their final concenttation in ceU culture medium. Volumes of the drug dUutions (lOO ⁇ L) were then added to each weU using a multichannel pipette. Plates were then mixed gently as above. The ceUs were incubated for a further 6 days at 37°C and 5% CO 2 . At this point the control weUs would have reached approximately 80-90% confluency.
  • COX-1 Cyclooxygenase-1 (COX-1) inhibition.
  • COX-1 is a constitutive enzyme and is thought to be associated with the important positive aspects of prostaglandins (cytoprotection and maintenance of renal blood flow).
  • Indomethacin is a potent inhibitor of COX-1 and as a result has a number of side effects associated with its use. Less inhibition of COX-1 is likely to be associated with fewer of these side effects.
  • test compounds were prepared as described previously.
  • the COX -1 assay was based on that used by Boopathy et al. 57 and Piazza et al. 58 .
  • the foUowrng compounds were weighed out and added to the lOOmM TRIS HCl: 0.05mM Glutathione 15.37mgs
  • the reaction mixture for the COX-1 assay was prepared as foUows:
  • TRIS-HCl + components (as prepared above) - added to make a final volume of lml in the reaction mixture. (Note : 250 units of COX-1 enzyme was used in the assay as gave the optimum level of activity when incubated with arachidonic acid.)
  • reaction mixture was incubated at 37°C for 20 minutes and then terminated by the addition of 0.2ml of 100% (w/v) ttichloroacetic acid in lm- HCl (91.7ml H 2 0 + 8.3ml Cone HCl ).
  • the enzymatic activity of COX-1 was assessed by its abUity to act on the substrate arachidonic acid and form malonaldehyde. The greater the optical density reading the greater the activity of COX-1 in the reaction mixture.
  • Indomethacin was used as the positive control.
  • the test compounds were added to the reaction mixture at the same concentration as indomethacin and their abUities to inhibit COX-1 were compared.
  • the controls used in the experiment were:
  • Arachidonic negative control Indomethacin negative control DMSO control. In the control reaction mixture arachidonic acid or enzyme was added to the reaction mixture after the addition the trichloroacetic acid.
  • results are the average of a minimum of three assay repeats. Data is expressed as % inhibition relative to an untreated control. Negative values indicate an increased activity of the enzyme in the presence of the compound. These results indicate that aU of the compounds tested (both active and inactive) have less COX-1 inhibitor activity and are therefore likely to have less side effects in the clinical setting.
  • Cyclooxygenase-2 (COX-2) is associated with the negative aspects of inflammatory conditions. Specific inhibition of this enzyme is thought to be therapeuticaUy useful in the treatment of chronic inflammatory conditions and may also have a beneficial role to play in the chemopreventation of certain form of colon cancer. Inhibition of COX-2, therefore, would be expected to provide additional useful effects in treatment using any of the positive compounds outlined.
  • test compounds were prepared as described previously.
  • COX-2-inhibitory activity was measured using an indirect method. It was previously shown by Asano et al., 59 , that COX-2 is the constitutive and dominant isoform in un-stimulated and stimulated cultured human lung epithelial ceUs. A lung adenocarcinoma ceU line, A549 was used as a source of COX-2. This ceU line was purchased from the American Type Culture CoUection, 10801 University Boulevard Manassas, VA 20110-2209, USA.
  • PGE 2 prostaglandin E 2
  • COX-2 interleukin l ⁇
  • the amount of PGE 2 produced is quantified by ELISA.
  • Inhibitors of COX-2 reduce the production of PGE 2 .
  • CeUs were seeded at high density (2.5 x 10 5 ceUs per weU) in 6 weU plates (Falcon, 3046).
  • Results are represented as means ⁇ S.D. for duplicate determinations carried out on three separate occasions. Inhibition is expressed as a percentage of untreated control (IL-l ⁇ (lOmg/ml)), taken as 100%.
  • indomethacin is also a potent inhibitor of COX-2 activity. Activity varies among the aU of compounds tested (both positive and negative) with Compound 4 having COX-2-inhibitory activity near to that of indomethacin.
  • GS-X pump is functionaUy overexpressed in cis-diamminedichloroplatinum(II)-resistant human leukemia HL-60 ceUs and down regulated by ceU differentiation. J.Biol
  • Apoptosis profundUy accounts for the growth inhibitory properties of sulindac metabolites and involves a mechanism that is independent of cyclooxygenase inhibition, ceU cycle arrest and p53 induction. Cancer Research, 57: 2452-2459.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Epidemiology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Indole Compounds (AREA)

Abstract

Cette invention concerne une famille de composés constituée de dérivés de l'acide acétique N-benzylindole-3, qui agissent en tant qu'inhibiteurs de protéine polypharmacorésistante (MRP) tout en présentant une faible toxicité directe, une faible activité inhibitrice de COX-1 et, dans certains cas, une action utile, notamment en ternes d'inhibition de COX-1. Ces agents sont utiles en combinaison avec des médicaments à substrat MRP pour le traitement de cancers pharmacorésistants ou de tumeurs risquant de se transformer en cancers pharmacorésistants dans la mesure où ils peuvent être administrés en concentrations plus fortes tout en étant moins toxiques et en ayant moins d'effets secondaires que les inhibiteurs de MRP connus. Ces agents peuvent également s'avérer utiles dans le traitement de maladies qui dépendent de l'activité de MRP-1 ou dans lesquelles l'activité de MRP-1 réduit l'efficacité des thérapies existantes.
PCT/IE2001/000109 2000-08-24 2001-08-24 Derives d'acide acetique n-benzylindole-3 pouvant etre utilises dans le traitement de cancers pharmacoresistants WO2002020478A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU2001282410A AU2001282410A1 (en) 2000-08-24 2001-08-24 N-benzylindole-3-acetic acid derivatives for use in the treatment of drug resistan cancer

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
EP00650115 2000-08-24
EP00650115.9 2000-08-24

Publications (1)

Publication Number Publication Date
WO2002020478A1 true WO2002020478A1 (fr) 2002-03-14

Family

ID=8174468

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/IE2001/000109 WO2002020478A1 (fr) 2000-08-24 2001-08-24 Derives d'acide acetique n-benzylindole-3 pouvant etre utilises dans le traitement de cancers pharmacoresistants

Country Status (3)

Country Link
AU (1) AU2001282410A1 (fr)
IE (1) IE20010784A1 (fr)
WO (1) WO2002020478A1 (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1744747A2 (fr) * 2004-04-26 2007-01-24 Vanderbilt University Derives d'acides indoleacetique et indenacetique comme agents therapeutiques a toxicite gastrointestinale reduite
US7736624B2 (en) 2006-06-19 2010-06-15 Univ Vanderbilt Methods and compositions for diagnostic and therapeutic targeting of COX-2
US9346803B2 (en) 2011-10-17 2016-05-24 Vanderbilt University Indomethacin analogs for the treatment of castrate-resistant prostate cancer

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1991006537A2 (fr) * 1989-10-27 1991-05-16 American Home Products Corporation Derives d'acide alcanoique indole, indene, pyranoindole et tetrahydrocarbazole substitues utilises comme inhibiteurs de la phospholipase a2 (pla2) et de la lipoxygenase
EP0620215A1 (fr) * 1993-04-16 1994-10-19 Eli Lilly And Company Dérivés de 1H-idole-3-acétamide comme inhibiteurs de sPLA2
EP0709090A2 (fr) * 1994-10-14 1996-05-01 Eli Lilly And Company Compositions pour le traitement de tumeurs résistantes
WO1998018490A1 (fr) * 1996-10-31 1998-05-07 Dublin City University Combinaisons destinees a augmenter la puissance d'un substrat pour une proteine associee a la resistance multiple aux medicaments
WO1998039330A1 (fr) * 1997-03-04 1998-09-11 Abbott Laboratories Composes heterocycliques inhibiteurs du cox-2

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1991006537A2 (fr) * 1989-10-27 1991-05-16 American Home Products Corporation Derives d'acide alcanoique indole, indene, pyranoindole et tetrahydrocarbazole substitues utilises comme inhibiteurs de la phospholipase a2 (pla2) et de la lipoxygenase
EP0620215A1 (fr) * 1993-04-16 1994-10-19 Eli Lilly And Company Dérivés de 1H-idole-3-acétamide comme inhibiteurs de sPLA2
EP0709090A2 (fr) * 1994-10-14 1996-05-01 Eli Lilly And Company Compositions pour le traitement de tumeurs résistantes
WO1998018490A1 (fr) * 1996-10-31 1998-05-07 Dublin City University Combinaisons destinees a augmenter la puissance d'un substrat pour une proteine associee a la resistance multiple aux medicaments
WO1998039330A1 (fr) * 1997-03-04 1998-09-11 Abbott Laboratories Composes heterocycliques inhibiteurs du cox-2

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
CHEMICAL ABSTRACTS, vol. 70, no. 17, 28 April 1969, Columbus, Ohio, US; abstract no. 76217s, BRASHER, W. JAMES ET AL.: "Effects of prednisolone, indomethacin, and Aloe vera gel on tissue culture cells." XP002161717 *
DATABASE HEMICAL ABSTRACTS XP002161718 *
EDWARD WALTON ET AL.: "Some analogs of 1-p-Chlorobenzyl-5-methylindole-3-acetic acid", JOURNAL OF MEDICINAL CHEMISTRY., vol. 11, - 1968, AMERICAN CHEMICAL SOCIETY. WASHINGTON., US, pages 1252 - 1255, XP002161716, ISSN: 0022-2623 *
ORAL SURG., ORAL MED., ORAL PATHOL., vol. 27, no. 1, - 1969, pages 122 - 128 *
ROBERT D. DILLARD ET AL.: "Indole inhibitors of human nonpancreatic secretory phospholipase A2. 2. Indole-3-acetamides with additional functionality", JOURNAL OF MEDICINAL CHEMISTRY., vol. 39, no. 26, - 1996, AMERICAN CHEMICAL SOCIETY. WASHINGTON., US, pages 5137 - 5158, XP002161715, ISSN: 0022-2623 *

Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1744747A2 (fr) * 2004-04-26 2007-01-24 Vanderbilt University Derives d'acides indoleacetique et indenacetique comme agents therapeutiques a toxicite gastrointestinale reduite
JP2007534702A (ja) * 2004-04-26 2007-11-29 バンダービルト・ユニバーシティ 胃腸毒性の低い治療薬としてのインドール酢酸、及びインデン酢酸誘導体
EP1744747A4 (fr) * 2004-04-26 2009-12-02 Univ Vanderbilt Derives d'acides indoleacetique et indenacetique comme agents therapeutiques a toxicite gastrointestinale reduite
US8168656B2 (en) 2004-04-26 2012-05-01 Vanderbilt University Indoleacetic acid and indenacetic acid derivatives as therapeutic agents with reduced gastrointestinal toxicity
US7736624B2 (en) 2006-06-19 2010-06-15 Univ Vanderbilt Methods and compositions for diagnostic and therapeutic targeting of COX-2
US8143302B2 (en) 2006-06-19 2012-03-27 Vanderbilt University Methods and compositions for diagnostic and therapeutic targeting of COX-2
US8865130B2 (en) 2006-06-19 2014-10-21 Vanderbilt University Methods and compositions for diagnostic and therapeutic targeting of COX-2
US9346803B2 (en) 2011-10-17 2016-05-24 Vanderbilt University Indomethacin analogs for the treatment of castrate-resistant prostate cancer
US9895351B2 (en) 2011-10-17 2018-02-20 Vanderbilt University Indomethacin analogs for the treatment of castrate-resistant prostate cancer
US10398678B2 (en) 2011-10-17 2019-09-03 Vanderbilt University Indomethacin analogs for the treatment of castrate-resistant prostate cancer
US11738004B2 (en) 2011-10-17 2023-08-29 Vanderbilt University Indomethacin analogs for the treatment of castrate-resistant prostate cancer

Also Published As

Publication number Publication date
AU2001282410A1 (en) 2002-03-22
IE20010784A1 (en) 2003-04-16

Similar Documents

Publication Publication Date Title
Guan et al. Expression of peroxisome proferator-activated receptor γ (PPARγ) in human transitional bladder cancer and its role in inducing cell death
Krysan et al. COX‐2‐dependent stabilization of survivin in non‐small cell lung cancer
US6756399B2 (en) Use of lipoxygenase inhibitors and PPAR ligands as anti-cancer therapeutic and intervention agents
AU730261B2 (en) Method of treating malignant tumors with thyroxine analogues having no significant hormonal activity
US20190314371A1 (en) Use of modulators of ccr5 in the treatment of cancer and cancer metastasis
JP3234818B2 (ja) 腫瘍性病変を抑制する化合物の同定方法
Aneja et al. Rational design of the microtubule-targeting anti–breast cancer drug EM015
JP2004512381A (ja) Ｈｓｐ９０インヒビターの使用により細胞毒性剤の効力を増強させる方法
WO2011035321A1 (fr) Traitement de maladies liées à wnt/frizzled
WO2012050985A1 (fr) Inhibiteurs du facteur sv40 tardif (lsf) en tant qu'agents chimio-thérapeutiques anticancéreux
EP1267874A2 (fr) Utilisations therapeutiques des mediateurs ppar
WO2006047716A2 (fr) Utilisation d'indoles lies au diindolylmethane et d'inhibiteurs du recepteur de facteur de croissance pour le traitement d'une maladie associee au cytomegalovirus humain
EP1328513A2 (fr) Compositions de diindolylmethane et de diindolylmethane c-substitue et procedes de traitement de cancers multiples
Kang et al. Expression of NSAID-activated gene-1 by EGCG in head and neck cancer: involvement of ATM-dependent p53 expression
US20040097408A1 (en) Compounds regulating cell proliferation and differentiation
Touhey et al. Structure–activity relationship of indomethacin analogues for MRP-1, COX-1 and COX-2 inhibition: identification of novel chemotherapeutic drug resistance modulators
KR20040079911A (ko) 리포좀에 의한 비타민 ε계 화합물의 전달
Ding et al. Ubiquitination of NOTCH2 by DTX3 suppresses the proliferation and migration of human esophageal carcinoma
Mahmud et al. Orally available tubulin inhibitor VERU-111 enhances antitumor efficacy in paclitaxel-resistant lung cancer
Bai et al. 5-(3, 4, 5-trimethoxybenzoyl)-4-methyl-2-(p-tolyl) imidazol (BZML) targets tubulin and DNA to induce anticancer activity and overcome multidrug resistance in colorectal cancer cells
Fang et al. Discovery and optimization of 2-(trifluoromethyl) benzimidazole derivatives as novel ferroptosis inducers in vitro and in vivo
WO2002020478A1 (fr) Derives d'acide acetique n-benzylindole-3 pouvant etre utilises dans le traitement de cancers pharmacoresistants
Kyriakou et al. Efficacy of cannabinoids against glioblastoma multiforme: A systematic review
JP2013518906A (ja) 複製タンパク質ａを阻害する物質および方法ならびにその使用
Keedwell et al. A retinoid-related molecule that does not bind to classical retinoid receptors potently induces apoptosis in human prostate cancer cells through rapid caspase activation

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BY BZ CA CH CN CO CR CU CZ DE DK DM DZ EC EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NO NZ PH PL PT RO RU SD SE SG SI SK SL TJ TM TR TT TZ UA UG US UZ VN YU ZA ZW

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): GH GM KE LS MW MZ SD SL SZ TZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE TR BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG

DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
121 Ep: the epo has been informed by wipo that ep was designated in this application
REG Reference to national code

Ref country code: DE

Ref legal event code: 8642

122 Ep: pct application non-entry in european phase
NENP Non-entry into the national phase

Ref country code: JP