Classifications

• • 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
  • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
  • A61K31/42—Oxazoles
• • 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
  • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
  • A61K31/19—Carboxylic acids, e.g. valproic acid
  • A61K31/192—Carboxylic acids, e.g. valproic acid having aromatic groups, e.g. sulindac, 2-aryl-propionic acids, ethacrynic acid 
• • 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
  • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
  • A61K31/19—Carboxylic acids, e.g. valproic acid
  • A61K31/195—Carboxylic acids, e.g. valproic acid having an amino group
  • A61K31/197—Carboxylic acids, e.g. valproic acid having an amino group the amino and the carboxyl groups being attached to the same acyclic carbon chain, e.g. gamma-aminobutyric acid [GABA], beta-alanine, epsilon-aminocaproic acid, pantothenic acid
  • A61K31/198—Alpha-aminoacids, e.g. alanine, edetic acids [EDTA]
• • 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
  • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
  • A61K31/425—Thiazoles
  • A61K31/426—1,3-Thiazoles
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P35/00—Antineoplastic agents

Definitions

• the invention relates to the use of a KMO inhibitor for the production of a pharmaceutical composition.
• KMO is a NADPH-dependent monooxygenase that catalyzes the reaction of L-kynurenine to L-3-hydroxykynurenine in tryptophan metabolism [see Bertazzo, A., Ragazzi, E., Biasiolo, M., Costa, C.V. and Allegri, G,; Enzyme activities involved in tryptophan metabolism along the kynurenine pathway in rabbits. Biochim Biophys Acta 1527 (2001) 167-75.]. KMO is also called kynurenine 3-monooxy-genase or kynurenine 3-hydroxylase
• Human KMO consists of 486 amino acids and is under accession no. NM__003679 known.
• the invention is based on the technical problem of specifying a pharmaceutical composition for the diagnosis and treatment of cancer.
• the invention teaches the use of a KMO inhibitor for the production of a pharmaceutical composition for the treatment of
• KMO-binding substance for the manufacture of a pharmaceutical composition for the detection of tumor cells.
• the invention is based on the knowledge that KMO is expressed or overexpressed in tumor cells compared to normal cells of the same tissue. On the one hand, this allows KMO to be used as a marker for identifying tumor cells.
• the inhibition of KMO offers the possibility, inter alia, to intervene in the tumor-related metabolism via NAD and thus ultimately to disrupt the tumor cell-specific metabolism and to contribute to the death or at least growth inhibition of the tumor cells.
• a KMO-binding substance is used in a pharmaceutical composition for the detection of tumor cells, it is preferred if the KMO-binding substance
• Substance is a KMO inhibitor.
• the uses according to the invention are particularly suitable for the treatment or diagnosis of breast cancer or uterine cancer.
• the inhibition can take place at the protein level or nucleic acid level. It is therefore preferred if the inhibitor is selected from the group consisting of a) inhibitor of the KMO protein, b) inhibitor of the KMO-RMA.
• the inhibitor of the KMO protein can be a molecule according to Formula I.
• 0 is a double bond and R2 is selected from: H, 2-C1, 2-N02, 3-Cl, 3-Br, 3- F, 4-C1, 4-Br, 4-F, 4-Me, 2,3-di-Cl, 2,3-di-F, 3,4-di-Cl, 3,4-di-F, 3-C1-4-F, 3-Me-4-Cl, 3-N02-4-C1.
• Z COOMe, COOH or a group selected from "furan, thiofuran, azole, isoazole, diazole, isodiazole, traizole, dithiol, oxathiol, isoxalzol, oxazole, thiazole, isothiazole, oxadiazole, tetrazole, oxatriazole, dioxazole, oxathiazole, thiatriazo'l
• the inhibitor of the KMO protein is a molecule of the formula II
• the inhibitor of the KMO protein can be an antibody against (human) KMO.
• Such antibodies can be polyclonal or monoclonal. They can be obtained in a customary manner by immunizing non-human mammals, for example rodents, with human KMO or with peptides which are specific to human KMO. However, it is not necessary to use a complete antibody, rather only the variable part can be used.
• Antibodies, complete or fragmentary can come from non-human organisms and can be humanized or represent chimeric antibodies. It is also possible to produce synthetic substances that mimic the binding region of an antibody, i.e. on the one hand have corresponding binding sites and on the other hand fix these binding sites in a defined spatial orientation to one another, corresponding to an antibody.
• an inhibitor of the KMO-RNA is used, which is selected from the group consisting of "ribozyme, aptamer and antisense nucleic acid against KMO".
• the embodiment of the invention is preferably used as a pharmaceutical composition for the treatment of cancer, in which KMO is specifically expressed or overexpressed by tumor cells.
• KMO is specifically expressed or overexpressed by tumor cells.
• a pharmaceutical composition according to the invention can be used to test for KMO dependency in vivo. If expression or overexpression of KMO compared to normal tissue of the same type is found, the use of the pharmaceutical composition according to the invention is indicated.
• the KMO-binding substance also carries a cytotoxic component. This ultimately leads to the fact that almost exclusively tumor cells are killed, while normal cells are almost completely retained in the tissue. If a cytotoxic component is used, it can be recommended that the pharmaceutical composition be prepared for local application in tissue containing tumor cells, for example for injection.
• Tumor cells preferably carry the KMO-binding substance additionally with a reporter group.
• a reporter group By using an assay tailored to the reporter group, it can be determined whether or to what extent KMO is expressed or overexpressed in cells of the examined tissue. In this way it can easily be determined whether tissue contains KMO-dependent tumor cells. It is also possible to determine whether one with other methods identified tumor tissue is KMO-dependent and is suitable for treatment with a pharmaceutical composition according to the invention.
• the invention relates to a method for diagnosing a cancer, a pharmaceutical composition according to the invention being applied in the embodiment with a reporter group to the tissue to be examined, the tissue to be examined then being subjected to a detection method step which is sensitive to the reporter group, and in the case of the detection of a defined minimum value of the reporter group in the tissue, the tissue is qualified as containing tumor cells and a method for the treatment of a tumor disease, a pharmaceutical composition according to the invention being given to a patient.
• the explanations given in this description apply analogously to these methods according to the invention.
• KMO is also called kynurenine 3-monooxygenase or kynurenine 3-hydroxylase.
• the protein is known under the accession number NM_003679.
• a KMO inhibitor is a compound or substance which either inhibits the formation of KMO or reduces the KMO activity formed, based on the KMO activity in the absence of the KMO inhibitor.
• a KMO inhibitor can be a substance that interferes with the KMO cascade.
• it can be a KMO inhibitor be a substance which forms a bond with the KMO formed, in such a way that the KMO can no longer catalyze the reaction from L-kynurenine to 3-hydroxykynurenine or only with a reduced conversion rate.
• mimicry molecules of L-kynurenine in particular are considered, but not the latter itself.
• Mimicry molecules are compounds which either completely prevent further reactions or lead to physiologically inactive catalytic products in the downstream region of the kynurenine pathway of tryptophan metabolism. Typical examples of this are shown in formula I.
• Counter ions for ionic compounds are, for example, Na + , K + , Li + or cyclohexylammonium.
• Suitable solid or liquid galenical forms of preparation are, for example, granules, powders, dragees, tablets, (micro) capsules, suppositories, syrups, juices, suspensions, emulsions, drops or injectable solutions (IV, IP, I.) and preparations with a protracted active ingredient -Release, in the production of which customary auxiliaries such as carriers, disintegrants, binders, coating agents, swelling agents, lubricants or lubricants, flavorings, sweeteners and solubilizers are used.
• customary auxiliaries such as carriers, disintegrants, binders, coating agents, swelling agents, lubricants or lubricants, flavorings, sweeteners and solubilizers are used.
• a pharmaceutical composition according to the invention can be produced in that at least one KMO inhibitor used according to the invention is mixed in a defined dose with a pharmaceutically suitable and physiologically compatible carrier and, if appropriate, further suitable active substances, additives or auxiliaries with a defined inhibitor dose and is prepared for the desired dosage form.
• An inhibitor of the KMO protein is a substance or compound which binds to KMO and reduces its activity towards KMO in the absence of the inhibitor.
• the KMO-RNA inhibitor refers to compounds or substances which inhibit transcription, bind RNA, cut if necessary, and / or inhibit translation.
• Substituted Cl-8-alkyl, aryl, or aralkyl carry a chemical group instead of one or more hydrogen atoms. Examples include OH, ether, ester, primary amine, secondary amine, tertiary amine, carboxyl, nitro and halogen.
• antibody encompasses both monoclonal antibodies and polyclonal antibodies.
• Tumor cells express KMO specifically when normal cells of the same tissue type do not express KMO. Tumor cells overexpress KMO specifically if KMO is expressed in at least double the amount compared to normal cells of the same tissue.
• Cytotoxic components or groups are compounds which directly or indirectly induce apoptosis or at least inhibit growth.
• Such groups or Compounds can be cytostatics in particular, which are used in tumor therapy. Examples of these are: alkylating agents (for example mechlorethamine, ifosfamide, chlorambucil, cyclophosphamide, melphalan, alkylsulfonates, butulfan, nitrosoureas, carmustine, lomustine, semustin, triazenes, dacarbazine), antimetabolites (for example folic acid antagonists, pyrimidine) -Analogues, fluorouracil, fluorodesoxyuridine, cytarabine, gemcitabine, purine analogues, mercaptopurine), mitosis inhibitors (e.g.
• vinca alkaloids voncristin, vinblastine, paclitaxal, docetaxel
• epipodophyllo-toxins e.g. etoposide, yeastipodin, eg, antibiotics, eg. Daunorubicin, idarubicin, anthracycline, bleomycin, L-asparaginase
• platinum complex compounds eg cisplatin
• hormones and related compounds eg adrenal gland steroids, aminogluthetides, progestogens, estrogens, androgens, anti-oestrogens, tamoxifen, steriodanalogens).
• the coupling takes place in such a way that the affinity for KMO is reduced by no more than 90%, preferably 50%, based on the substance without a cytostatic group, and the cytostatic effect of the group is not reduced by more than 90%, preferably 50%, based on the compound without substance.
• a reporter group is an atom, molecule or a compound which, in conjunction with an assay placed thereon, enables the detection of the reporter group and the compound or substance thus connected to the reporter group.
• reporter groups and associated detection methods are: 32P labeling and intensity measurement using phosphoimager. Many other examples are known to the average person skilled in the art and do not require a detailed list.
• a substance that binds to KMO can be a substance that binds a KMO protein or KMO RNA.
• the KMO coding sequence was labeled with 32P by means of the rando hex priming and hybridized to a "Cancer profiling array" from Clontech, which contains 240 cDNA library pairs, each pair comprising tumor and Represents normal tissue from a single patient.
• the signal intensities obtained were determined using a phosphor imager and the tumor / normal ratio calculated for each pair. The results obtained are shown in FIG. 1. It can be seen that 18 out of 53, ie 34%, of the breast tumors show at least two-fold overexpression of KMO. Within the subgroup of invasive ductal carcinomas, 43% of the tumors showed at least two-fold overexpression.
• the KMO coding sequence was labeled with 32P by means of the randorn hexmer priming and hybridized to a "cancer profiling array" from Clontech which contains 240 cDNA library pairs, each pair representing tumor and normal tissue from a single patient.
• the signal intensities obtained were quantified using a phosphor imager and tumor / normal ratios were calculated.
• FIG. 2. 20 of 43, i. e. 46% of the uterine organs showed at least two-fold overexpression of KMO. Some of the tumors even showed more than tenfold overexpression, the maximum being almost thirtyfold in uterine adenocarcinoma.
• FIG. 4 Various compounds are shown in FIG. 4 which are suitable as inhibitors of the KMO protein. Because of their structure related to L-kynurenine, these compounds bind to KMO competitively with L-kynurenin and thus block the reaction of L-kynurenine to 3-hydroxykynurenine in the cell.
• Seq-ID 1 shows a usable antisense nucleic acid (phosphothioate oligo, 20mer). This binds to KMO RNA and inhibits translation.
• Polyclonal KMO antibodies were generated in Gallus gallus by immunization with one or more of the KMO-specific peptides of the sequences Seq-ID 6-8, which had been conjugated with a carrier protein. Sera were removed and the antibodies contained therein were affinity purified using the immobilized three peptides mentioned.
• a radioactively labeled probe was used for hybridization, which covers the complete coding region of the KMO gene, but does not contain any region which could cross-react with other genes.
• An array commercially available from BioCat was used, which was created using tissue samples taken from human patients. Such an array contains thin sections from 55 different adenocarcinomas of the prostate as well as 2 tissue sections from normal prostate tissue.
• radioactively labeled probe was used for hybridization, which covers the complete coding region of the KMO gene, but does not contain any region which could cross-react with other genes.
• Example 9 Cell culture-based assay with tumor cell lines
• nude mice are each given 10 6 cells of the prostate tumor cell line PC3. This can be done by subcutaneous injection or by orthotopic injection into the prostate. 3 days after the injection, some of the animals (rest: control group) were given a defined dose of the inhibitor intraperitoneally, for example 400 mg / kg / day, daily.
• the influence of the inhibitor on tumor development is quantified by comparing the tumor growth rates between the test group and the control group. The first orienting results indicate that an inhibition of proliferation also takes place in vivo.

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• 2001
  • 2001-11-28 DE DE10159215A patent/DE10159215A1/de not_active Ceased
• 2002
  • 2002-11-20 AU AU2002351693A patent/AU2002351693A1/en not_active Abandoned
  • 2002-11-20 WO PCT/DE2002/004309 patent/WO2003047571A2/de not_active Application Discontinuation

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