WO2013144894A1 - Analogue de nucléoside utilisable en tant que composé anticancéreux - Google Patents

Analogue de nucléoside utilisable en tant que composé anticancéreux Download PDF

Info

Publication number
WO2013144894A1
WO2013144894A1 PCT/IB2013/052481 IB2013052481W WO2013144894A1 WO 2013144894 A1 WO2013144894 A1 WO 2013144894A1 IB 2013052481 W IB2013052481 W IB 2013052481W WO 2013144894 A1 WO2013144894 A1 WO 2013144894A1
Authority
WO
WIPO (PCT)
Prior art keywords
cancer
compound
formula
leukemia
pharmaceutically acceptable
Prior art date
Application number
PCT/IB2013/052481
Other languages
English (en)
Inventor
Abhijeet Sudhir KATE
Saji David George
Shailendra SONAWANE
Giridharan Periyasamy
Original Assignee
Piramal Enterprises Limited
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 Piramal Enterprises Limited filed Critical Piramal Enterprises Limited
Publication of WO2013144894A1 publication Critical patent/WO2013144894A1/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
    • C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
    • C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
    • C07H19/06—Pyrimidine radicals
    • 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
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/26—Preparation of nitrogen-containing carbohydrates
    • C12P19/28—N-glycosides
    • C12P19/38—Nucleosides
    • C12P19/385—Pyrimidine nucleosides
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/04—Actinomyces

Definitions

  • the present invention relates to a compound of formula (1) (as described herein), or a pharmaceutically acceptable salt or a derivative thereof.
  • the compound may be obtained by fermentation of a microorganism belonging to marine actinomycetes strain (PM0895117/MTCC 5675).
  • the present invention further relates to processes for the production of the compound of formula (1), to the microorganism belonging to marine actinomycetes strain (PM0895117/MTCC 5675) and to pharmaceutical compositions containing the compound of formula (1) as an active ingredient and use thereof for the treatment of cancer.
  • Cancer has become an increasing public health problem due to its high rates of morbidity and mortality. Despite continuing advances in treatment regimens for cancer, this disease still remains one of the leading causes of death in the world for the reason that the available treatment options are associated with undesirable side effects and limited efficacy. It is the second most common cause of death in developed countries, exceeded only by heart diseases. Cancer accounts for nearly one of every four deaths (Source: Cancer Facts and Figures 2010 of the American Cancer Society). By the year 2020, cancer could kill more than ten million people per year unless action is taken in both the field of prevention and treatment (Biotechnology Advances, 29, 2011, 531-547).
  • Cancer therapy currently falls under the following categories including surgery, radiation therapy, chemotherapy, bone marrow transplantation, stem cell transplantation, hormonal therapy, immunotherapy, antiangiogenic therapy, targeted therapy, gene therapy and others.
  • the marine environment covering seventy percent of the earth's surface and ninety five percent of its tropical biosphere represents thirty four of the thirty six phyla of life and provides a beautiful variety of biodiversity exceeding that of the terrestrial environment (Life Sciences, 78, 2005, 442 ⁇ -53).
  • Marine natural product bio prospecting which is exploration, extraction and screening of biological diversity and indigenous knowledge for commercially valuable genetic and biochemical resources, has yielded a considerable number of drug candidates (Drug Discovery Today, 8 (12), 2003, 536-544). Marine natural products have become spectacular targets for biologists and chemists for discovery of lead compounds for clinical development since the past five decades (Evidence-Based Complementary and Alternative Medicine, 2011, 2011, 1-11).
  • anticancer agents have been discovered by screening natural products from marine organisms and microorganisms.
  • anticancer agents originating from marine sources include citarabine, bryostatin-1, aplidine, dolastatin 10 and ET-743. (Current Opinion in Pharmacology, 1, 2001, 364—369).
  • Plicacetin is a nucleoside antibiotic isolated from actinomycete Streptomyces plicatus.
  • Cytosaminomycin A is a nucleoside anticoccidial antibiotic isolated from the fermentation broth of Streptomyces amakusaensis KO-81 191 (The Journal Of Antibiotics, 47 (7), 1994, 774-781, and 47 (7), 1994, 782-786). In view of this, it would be prudent to explore this area of research to identify and develop new drugs which can be used effectively to treat various types of cancers.
  • the inventors of the present invention have directed their efforts in carrying out an extensive research in this area which resulted in the present invention.
  • the present invention relates to a nucleoside analogue designated herein as compound of formula (1) (as described herein).
  • the present invention also relates to a purified compound of formula (1), isolated from the fermented broth of the marine actinomycetes strain (PM0895117/MTCC 5675) or one of its variants or mutants.
  • the invention also relates to all isomeric forms and tautomeric forms of the compound of formula (1), and a pharmaceutically acceptable salt or a derivative thereof.
  • the compound of formula (1) or an isomer, a tautomer,or a pharmaceutically acceptable salt or a derivative thereof, for use in the treatment of cancer.
  • the invention further relates to a pharmaceutical composition
  • a pharmaceutical composition comprising the compound of formula (1), or an isomer, a tautomer, a pharmaceutically acceptable salt, or a derivative thereof, as an active ingredient in association with at least one pharmaceutically acceptable excipient, carrier or a vehicle.
  • the present invention also relates to processes for the production of the compound of formula (1) and/or its isomers or its tautomers from the marine actinomycetes strain (PM0895117/MTCC 5675).
  • the present invention also relates to processes for the isolation of the marine actinomycetes strain (PM0895117/MTCC 5675), which on cultivation produces the compound of formula (1), or its isomers or tautomers.
  • the present invention relates to a method for the treatment of cancer in a subject comprising administering to the subject a therapeutically effective amount of a compound of formula (1), or an isomer or a tautomer or a pharmaceutically acceptable salt or a derivative thereof.
  • Figure 1 illustrates H NMR spectrum (DMSO-d 6 ; 500 MHz; Instrument: Bruker) of the compound of formula (1).
  • Figure 2 illustrates 13 C NMR spectrum (DMSO-d 6 ; 75 MHz; Instrument: Bruker) of the compound of formula (1).
  • Figure 3 illustrates effect of compound of formula (1) on apoptosis in Panc-1 cancer cells.
  • the compound of formula (1) has the molecular formula C 22 H 34 N 4 O 7 S and a molecular weight 498.2.
  • the compound of formula (1) may be characterized by using any one or more of the physico-chemical and spectral methods, such as high performance liquid chromatography (HPLC), mass spectrometry (MS), infra red spectroscopy (IR), and nuclear magnetic resonance (NMR) spectroscopic data as discussed herein below.
  • HPLC high performance liquid chromatography
  • MS mass spectrometry
  • IR infra red spectroscopy
  • NMR nuclear magnetic resonance
  • the compound of formula (1) is structurally represented as follows:
  • the gram-positive bacteria which may be used for the production of the compound of formula (1), is a marine actinomycetes strain (PM0895117/MTCC 5675), herein after referred to as culture no. PM0895117, which was isolated from the deep sea sediment collected from the offshore region of Mumbai, Maharashtra, India.
  • One aspect of the present invention provides processes for the production of the compound of formula (1) from the culture no. PM0895117 comprising the steps of:
  • step (c) involving purification of the compound of formula (1) is carried out by purification procedures generally used in the related art.
  • the compound of formula (1) produced according to the process of the present invention is a substantially pure compound.
  • the compound of formula (1) is an isolated pure compound produced from the culture no. PM0895117.
  • mutant refers to an organism or cell carrying a mutation, which is an alternative phenotype to the wild-type.
  • variable refers to an individual organism that is recognizably different from an arbitrary standard type in that species.
  • mammal refers to a human as well as non-human mammal, including but not limited to, cows, horses, pigs, dogs and cats.
  • mammal may be used interchangeably with the term “patient” or "subject”.
  • whole broth may be used interchangeably with the terms "nutrient broth”, “culture broth” or “fermented broth”.
  • active ingredient refers to the compound of formula (1) or to an isomer or a tautomer or a derivative or a pharmaceutically acceptable salt thereof.
  • compound of formula (1) includes the compound of formula (1) itself and an isomer, a tautomer, or a pharmaceutically acceptable salt thereof.
  • substantially pure means that the compound of formula (1) or an isomer thereof is sufficiently pure such that further purification would not detectably alter its physical and chemical properties, such as enzymatic and biological activities, of the substance.
  • the compound of formula (1) can be purified substantially by following the methods known to those skilled in the art.
  • the term "therapeutically effective amount” in reference to the treatment of cancer (as listed herein) using the compound of formula (1), a pharmaceutically acceptable salt thereof or a derivative thereof, refers to an amount capable of invoking one or more of the following effects in a subject receiving the compound of the present invention: (i) inhibition, to some extent, of tumor growth, including, slowing down and complete growth arrest; (ii) reduction in the number of tumor cells; (iii) reduction in tumor size; (iv) inhibition (i.e., reduction, slowing down or complete stopping) of tumor cell infiltration into peripheral organs; (v) inhibition (i.e., reduction, slowing down or complete stopping) of metastasis; (vi) enhancement of anti-tumor immune response, which may, but does not have to, result in the regression of the tumor; and/or (vii) relief, to some extent, of one or more symptoms associated with the cancer being treated.
  • salt(s) means those salts of compound(s) of the invention that are safe and effective in mammals and that possess the desired biological activity.
  • Pharmaceutically acceptable acid addition salts include, but are not limited to, hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, phosphate, acetate, lactate, salicylate, citrate, tartrate, ascorbate, succinate, maleate, fumarate, formate, benzoate, glutamate, methanesulfonate, benzensulfonate, or p-toluenesulfonate salts.
  • Suitable base addition salts include, but are not limited to, calcium, lithium, magnesium, potassium, sodium, or zinc, salts.
  • mutants of PM0895117 such as those produced by the use of chemical or physical mutagens including X-rays, U.V. rays etc. and organisms whose genetic makeup has been modified by molecular biology techniques, may also be cultivated to produce the compound of formula (1).
  • the screening for suitable mutants and variants which can produce the compound according to the invention can be confirmed by HPLC, NMR, IR, MS and/or determination of biological activity of the active compounds accumulated in the culture broth, for example by testing the compounds for anticancer activity or by a combination thereof.
  • the medium and/or nutrient medium used for isolation and cultivation of culture no. PM0895117, which produces the compound of formula (1), preferably contains sources of carbon, nitrogen and nutrient inorganic salts.
  • the carbon sources are, for example, one or more of soluble starch, glucose, sucrose, dextrin, fructose, molasses, glycerol, lactose, or galactose.
  • a preferred carbon source is soluble starch and glucose.
  • the sources of nitrogen are, for example, one or more of soyabean meal, peanut meal, yeast extract, beef extract, peptone, malt extract, corn steep liquor, gelatin, sodium caseinate, L-Asparagine or casamino acids.
  • Preferred nitrogen source is soyabean meal, peptone, sodium caseinate, L-Asparagine, corn steep liquor and yeast extract.
  • the nutrient inorganic salts are, for example, one or more of sodium chloride, potassium chloride, calcium chloride, manganese chloride, magnesium chloride, strontium chloride, cobalt chloride, potassium bromide, sodium fluoride, sodium hydrogen phosphate, potassium hydrogen phosphate, dipotassium hydrogen phosphate, disodium phosphate, calcium carbonate, sodium propionate, sodium bicarbonate, sodium silicate, sodium nitrate, ammonium nitrate, potassium nitrate, sodium sulphate, ammonium sulphate, ammonium heptamolybdate, ferric citrate, copper sulphate, magnesium sulphate, ferrous sulphate, zinc sulphate or boric acid.
  • culture no. PM0895117 may be carried out at a temperature ranging from 24°C to 32°C. Typically, culture no. PM0895117 is maintained at 27°C - 29°C. The well-grown cultures may be preserved in the refrigerator at 4°C - 8°C.
  • Seed culture cultivation of culture no. PM0895117 may be carried out at a temperature ranging from 27°C to 33°C and a pH of about 5.5 to 8.5, for 60-80 hrs at 210- 260 rpm (revolutions per minute).
  • culture no. PM0895117 seed is cultivated at 29 °C - 31 °C and a pH of about 6.5- 7.5, for 70-74 hrs at 230-250 rpm.
  • the production of the compound of formula (1) may be carried out by cultivating culture no. PM0895117 by fermentation in shake flasks at a temperature ranging from 27°C to 33°C and a pH of about 5.5 to 8.5, for 80-110 hrs at 200- 250 rpm.
  • culture no. PM0895117 is cultivated at 29°C-31°C and pH 6.5- 7.5, for 90-100 hrs at 210- 230 rpm.
  • the production of the compound of formula (1) may be carried out by cultivating culture no. PM0895117 by fermentation in a fermenter at a temperature ranging from 27°C to 33°C and a pH of about 5.5 to 8.5, for 80-110 hrs at 40- 70 rpm and 400- 500 1pm (liters per minute) aeration.
  • culture no. PM0895117 is cultivated at 28°C-31°C and pH 6.5- 7.5, for 90-100 hrs at 50- 60 rpm and 435- 465 1pm aeration.
  • the production of the compound of formula (1) can be carried out by cultivating culture no. PM0895117 in a suitable nutrient broth under conditions described herein, preferably under submerged aerobic conditions in shake flasks or fermenters.
  • the progress of fermentation and production of the compound of formula (1) can be detected by high performance liquid chromatography (HPLC) and by measuring the bioactivity of the culture broth by testing against the cancer cell lines.
  • HPLC high performance liquid chromatography
  • Fermentation is a process of growing microorganisms for the production of various chemical or pharmaceutical compounds. Microbes are normally incubated under specific conditions in the presence of nutrients. Whole broth is obtained after completing the process of fermentation. The whole broth is subjected to centrifugation which results in formation of cell mass and culture filtrate, which can be processed further by processes, described herein.
  • the compound of formula (1) present in the culture broth may be isolated using different extraction methods and chromatographic techniques.
  • the compound of formula (1) can be recovered from the culture filtrate by extraction with a water immiscible solvent such as petroleum ether, dichloromethane, chloroform, ethyl acetate, diethyl ether or butanol, or by hydrophobic interaction chromatography using polymeric resins such as "Diaion HP-20 ® “ (Mitsubishi Chemical Industries Limited, Japan), “Amberlite XAD ® “ (Rohm and Haas Industries, USA) or adsorption on activated charcoal. These techniques may be used repeatedly, alone or in combination.
  • a water immiscible solvent such as petroleum ether, dichloromethane, chloroform, ethyl acetate, diethyl ether or butanol
  • polymeric resins such as "Diaion HP-20 ® " (Mitsubishi Chemical Industries Limited, Japan), “Amberlite XAD ® “ (Rohm and Haas Industries, USA) or adsorption on activated charcoal.
  • the compound of formula (1) can be recovered from the cell mass by extraction with a water miscible solvent such as methanol, acetone, acetonitrile, n-propanol, or iso-propanol or with a water immiscible solvent such as petroleum ether, dichloromethane, chloroform, ethyl acetate or butanol.
  • a water miscible solvent such as methanol, acetone, acetonitrile, n-propanol, or iso-propanol
  • a water immiscible solvent such as petroleum ether, dichloromethane, chloroform, ethyl acetate or butanol.
  • the whole broth may be extracted with a solvent selected from petroleum ether, dichlorome thane, chloroform, ethyl acetate, methanol, acetone, acetonitrile, n-propanol, iso-propanol, or butanol.
  • the compound of formula (1) is extracted from the whole broth using ethyl acetate. Concentration of the extract gives the active crude material.
  • the compound of formula (1) of the present invention can be recovered from the crude material by fractionation using any of the following techniques: normal phase chromatography (using alumina or silica gel as stationary phase; eluents such as petroleum ether, ethyl acetate, chloroform, dichloromethane, acetone, methanol, or combinations thereof; and if required, additions of amines such as triethylamine); reverse phase chromatography (using reverse phase silica gel such as dimethyloctadecylsilylsilica gel, (RP- 18) or dimethyloctylsilyl silica gel (RP-8) as stationary phase; and eluents such as water, buffers (for example, phosphate, acetate, citrate (pH 2-8)), and organic solvents (for example methanol, acetonitrile, acetone, tetrahydrofuran, or combinations of these solvents)); gel permeation chromatography (using resins such as Sep
  • isomer is a general term used for all isomers of the compound of formula (1) or of the derivative thereof that differ only in the orientation of their atoms in space.
  • isomer includes mirror image isomers (enantiomers), mixtures of mirror image isomers (racemates, racemic mixtures) and isomers of compounds with more than one chiral center that are not mirror images of one another (diastereoisomers).
  • the compound of the present invention may have asymmetric centers and occur as racemates, racemic mixtures, individual diastereoisomers, or enantiomers, or may exist as geometric isomers, with all isomeric forms of said compound being included in the present invention.
  • tautomer refers to the coexistence of two (or more) compounds that differ from each other only in the position of one (or more) mobile atoms and in electron distribution, for example, keto-enol tautomers.
  • the compound of formula (1), an isomer or a tautomer thereof can be converted into their pharmaceutically acceptable salts and derivatives, such as esters and ethers, which are all contemplated by the present invention.
  • salts can be prepared by standard procedures known to one skilled in the art, for example, salts like sodium and potassium salts, can be prepared by treating the compound of formula (1), or an isomer, a tautomer or a derivative thereof, with a suitable sodium or potassium base, for example sodium hydroxide or potassium hydroxide.
  • salts like hydrochloride and sulphate salts can be prepared by treating the compound of formula (1), or an isomer, a tautomer, and a derivative thereof, with a suitable acid, for example hydrochloric acid or sulphuric acid.
  • the derivative of the compound of formula (1) is preferably an ester or an ether of the compound of formula (1).
  • the esters and ethers of the compound of formula (1) can be prepared by the methods given in the literature (Advanced Organic Chemistry, 1992, 4 th Edition, J. March, John Wiley & Sons) and known by one skilled in the art.
  • the compound of formula (1) or a pharmaceutically acceptable salt or a derivative thereof has anticancer activity against a wide range of cancer cells.
  • the compound of formula (1), or an isomer, a tautomer, a pharmaceutically acceptable salt or a derivative thereof can be administered to a subject in need thereof as a pharmaceutical and in the form of a pharmaceutical composition.
  • the compound of formula (1) or an isomer, a tautomer, a pharmaceutically acceptable salt or a derivative thereof can be administered to a subject who is diagnosed having cancer.
  • the present invention also relates to the compound of formula (1), to the use of the compound of formula (1), or an isomer, a tautomer, a pharmaceutically acceptable salt or a derivative thereof for the manufacture of a medicament for the treatment of cancer.
  • the present invention further relates to a pharmaceutical composition
  • a pharmaceutical composition comprising a therapeutically effective amount of the compound of formula (1) or an isomer or a tautomer or a pharmaceutically acceptable salt or a derivative such as an ester or ether thereof, and a pharmaceutically acceptable excipient or a carrier.
  • the pharmaceutical composition is provided for use in the treatment of cancer.
  • the therapeutically effective amount of the compound of formula (1), or its stereoisomer, or its tautomer or its pharmaceutically acceptable salt or its derivative as the active ingredient in the pharmaceutical preparations may range from about 0.01 mg to 1000 mg.
  • the compounds of the present invention find use in the treatment of cancers.
  • Compounds of present invention are used to reduce, inhibit, or diminish the proliferation of tumor cells, and thereby assist in reducing the size of a tumor.
  • Representative cancers that may be treated by such compounds include but are not limited to leukemia, lung cancer, brain tumors, non-Hodgkin's lymphoma, Hodgkin's disease, liver cancer, kidney cancer, bladder cancer, cancer of urinary tract, breast cancer, head and neck cancer, endometrial cancer, lymphoma, melanoma, cervical cancer, thyroid cancer, gastric cancer, germ cell tumor, cholangiocarcinoma, extracranial cancer, sarcoma, mesothelioma, malignant fibrous histiocytoma of bone, retinoblastoma, esophageal cancer, multiple myeloma, head and neck cancer, pancreatic cancer, ependymoma, neuroblastoma, skin cancer,
  • the cancer is selected from acute lymphocytic leukemia, acute myeloid leukemia, adult acute myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, hairy cell leukemia, non-small-cell lung cancer, small-cell lung cancer, brain stem glioma, glioblastoma, astrocytoma including cerebellar astrocytoma and cerebral astrocytoma, visual pathway glioma, hypothalamic glioma, supratentorial primitive neuroectodermal, pineal tumors, medulloblastoma, primary central nervous system lymphoma, mantle cell lymphoma, Hodgkin's disease, hepatocellular carcinoma, renal cell carcinoma, Wilms' tumor, bladder cancer, cancer of urinary tract, Ewing's sarcoma family of tumors, osteosarcoma, rhabdomyosarcoma
  • the present invention provides a method for the treatment of cancer by administering to a mammal in need thereof a therapeutically effective amount of a compound of formula (1).
  • the compound of formula (1) or a pharmaceutically acceptable salt or a derivative thereof can be administered orally, nasally, topically, subcutaneously, intramuscularly, intravenously, or by other modes of administration.
  • the method of administration which is suitable in a specific case depends on the type of cancer to be treated and on the stage of the cancer. Further, the method of administration can be optimized by a medical practitioner using methods known in the art.
  • the dosage range which are suitable in a specific case depend on the type of cancer to be treated and on the state of the respective condition or disease, and can be optimized using methods known in the art.
  • the daily dose of active compound (the compound of formula (1)) in a patient is 0.05 mg to 200 mg per kg, typically 1 mg to 100 mg per kg.
  • compositions which contain compound of formula (1) or an isomer or a tautomer, a pharmaceutically acceptable salt or a derivative thereof can be prepared by mixing the compound of formula (1) with one or more pharmacologically tolerated auxiliaries and/or excipients such as, wetting agents, solubilisers such as surfactants, vehicles, tonicity agents, fillers, colorants, masking flavors, lubricants, disintegrants, diluents, binders, plasticizers, emulsifiers, ointment bases, emollients, thickening agents, polymers, lipids, oils, cosolvents, complexation agents, or buffer substances, and converting the mixture into a suitable pharmaceutical form such as, for example, tablets, coated tablets, capsules, granules, powders, creams, ointments, gels, syrup, emulsions, suspensions, or solutions suitable for parenteral administration.
  • auxiliaries and/or excipients such as, wetting agents
  • auxiliaries and/or excipients examples include cremophor, poloxamer, benzalkonium chloride, sodium lauryl sulphate, dextrose, glycerin, magnesium stearate, polyethylene glycol, starch, dextrin, lactose, cellulose, carboxymethylcellulose sodium, talc, agar-agar, mineral oil, animal oil, vegtetable oil, organic and mineral waxes, paraffin, gels, propylene glycol, benzyl alcohol, dimethylacetamide, ethanol, polyglycols, Tween 80, solutol HS 15, and water. It is also possible to administer the compound of formula (1) as such, without vehicles or diluents, in a suitable form, for example, in capsules.
  • the pharmaceutical compositions which contain compound of formula (1) or an isomer or a tautomer, a pharmaceutically acceptable salt or a derivative thereof may be used in combination with one or more anticancer agents such as asparaginase, bleomycin, carboplatin, carmustine, chlorambucil, cisplatin, colaspase, cyclophosphamide, cytarabine, dacarbazine, dactinomycin, daunorubicin, doxorubicin, epirubicin, etoposide, fluorouracil, hexamethylmelamine, hydroxyurea, ifosfamide, leucovorin, lomustine, mechlorethamine, 6-mercaptopurine, mesna, methotrexate, mitomycin C, mitoxantrone, prednisolone, prednisone, procarbazine, streptozocin, tamoxifen
  • anticancer agents such as
  • Agar medium sodium caseinate 0.2%, L- asparagine 0.01%, sodium propionate 0.4%, dipotassium hydrogen phosphate 0.05%, magnesium sulphate 0.01%, ferrous sulphate 0.0001%, glucose 1% and agar 1.5%; demineralized water ; pH 8.1.
  • ASW artificial seawater
  • genomic DNA was isolated from the sediment samples and amplified for the presence of polyketide synthase (PKS), non- ribosomal peptide synthetases (NRPS) and glycosyl transferase biosynthetic pathways by performing PCR. Culture of interest was then isolated from the sediment sample having Glycosyl transferase pathway.
  • PKS polyketide synthase
  • NRPS non- ribosomal peptide synthetases
  • Glycosyl transferase pathway was then isolated from the sediment sample having Glycosyl transferase pathway.
  • the sample was stored at -20°C to -22°C and later thawed to room temperature
  • Agar medium sodium caseinate 0.2%, L- asparagine 0.01%, sodium propionate 0.4%, dipotassium hydrogen phosphate 0.05%, magnesium sulphate 0.01%, ferrous sulphate 0.0001%, glucose 1% and agar 1.5%; demineralized water; pH 8.1.
  • the culture no. PM0895117 was streaked on a petriplate containing modified Actinomycetes Isolation Agar. The petriplate was incubated for ten days at 27°C. One of the isolated colonies from the petriplate was transferred to slants containing ISP4 Agar medium (HiMedia, India). The slants were incubated for ten days at 27°C.
  • This particular strain was maintained on slants containing ISP4 media (HiMedia, India). The media was dissolved thoroughly by heating; the resultant solution was distributed in test tubes and sterilized at 121°C for 30 minutes. The test tubes were cooled and allowed to solidify in a slanting position. The agar slants were streaked with well grown culture no. PM0895117, by a wire loop and incubated at 27°C to 29°C until a significant growth was observed. The well-grown cultures were stored in the refrigerator at 4°C to 8°C.
  • MBA8 medium calcium carbonate 0.2%, sodium chloride 0.5%, corn steep liquor 0.5%, peptone 0.75%, glucose 1.5% and yeast extract 0.75%; demineralized water to make up the volume up to 100 ml; pH-7.0.
  • Example 4 (a) 50 ml of the seed medium as prepared in Example 4 (a) was distributed in 500 ml capacity Erlenmeyer flasks and autoclaved at 121°C for 30 minutes. The flasks were cooled to room temperature (25 ⁇ 2°C) and each flask was inoculated with a loopful of the well- grown producing strain (culture no. PM895117) on the slant and shaken on a rotary shaker for 72 hrs at 230 rpm to 250 rpm at 30°C ( ⁇ 1°C) to give seed culture.
  • a loopful of the well- grown producing strain culture no. PM895117
  • MBA8 medium calcium carbonate 0.2%, sodium chloride 0.5%, corn steep liquor 0.5%, peptone 0.75%, glucose 1.5% and yeast extract 0.75%; demineralized water to make up the volume up to 1 L; pH-7.0. d) Procedure:
  • the production flasks were incubated on shaker at 30°C and 220 rpm for 96 hrs.
  • the production flasks were harvested (harvest pH: 7.0 to 8.0) and the whole broth from each media flask was extracted with equal volume of solvent mixture (1 :9 methanol: ethyl acetate). These flasks were kept at room temperature for 4-6 hrs for extraction followed by separation of the supernatant. The supernatant was used for testing for anti cancer activity.
  • Example 5 450 L of the production medium in 1KL fermenter along with 180 ml of Desmophen as an antifoaming agent was sterilized in situ for 30 minutes at 121°C, cooled to 29-30°C and seeded with 8-10 L of the seed culture as obtained in Example 5 (b).
  • the production fermenter was incubated at 29°C -30°C, agitated at 54 rpm and aeration of 450 1pm for 96 hrs.
  • the harvest pH of the culture broth was 6.5-7.5.
  • the production of the compound of formula (1) in the fermentation broth was determined by HPLC and by testing for anti cancer activity.
  • the harvested whole broth (450 L) from the Example 6 was extracted using ethyl acetate (450 L).
  • the organic layer was separated using disc stack separator (Alfa Laval Sweden), model no LAPX404) and concentrated to obtain the crude extract (600 g).
  • MCF-IOA non- tumourigenic cell line was cultured in Mammary Epithelial Basal Medium (MEBM) with all standard additions (Lonza, Catalog. No. CC-3150). All cells were grown at 37°C with 5% CO 2 incubator. Cells were passaged at 80 - 90% confluence. Adherent cells were trypsinised using Trypsin-EDTA (Sigma) and maintained. Step 2
  • Compound of formula (1) was dissolved in DMSO to give a required stock solution of 20 mM. Eight different concentrations of the compound of formula (1) were prepared by serial dilution of the stock solution finally resulting in a 200-fold higher concentration (compared to the test concentration). Compound of formula (1) was tested at the concentrations of 30 ⁇ , 10 ⁇ , 3 ⁇ , 1 ⁇ , 0.3 ⁇ , 0.1 ⁇ , 0.03 ⁇ and 0.01 ⁇ . Each concentration was evaluated in triplicate.
  • 200 X 200 times higher than required concentration is denoted as 200 X
  • compound of formula (1) dissolved in DMSO was added to the above tissue culture plate seeded with cancer/normal cells.
  • the final concentration of DMSO was 0.5% in wells.
  • DMSO was used as a vehicle control.
  • the annexinV assay a classical technique for detecting apoptosis, is the most commonly used method for detecting apoptosis by flow cytometry.
  • One of the earliest features of apoptosis is the translocation of phosphatidylserine from the inner to the outer leaflet of the plasma membrane, thereby exposing phosphatidylserine to the external environment.
  • AnnexinV binds to phosphatidylserine exposed on the cell surface and identifies cells at an earlier stage of apoptosis.
  • Panc-1 cells [referred to in step 1 of example 8] were seeded at a density of 0.5 x 10 6 cells/2000 ⁇ ⁇ well, in a tissue culture grade 96 well plate and allowed them to recover for 24 hrs in a humidified 5 ⁇ 0.2% C0 2 incubator at 37 ⁇ 0.5 °C.
  • the processed cells were analyzed in BD flow cytometer for annexin V detection.
  • the population present in Ml and M2 were determined using FACS analysis.
  • Protein expressions of p53 (Santa cruz biotechnology, Inc. USA, Catelog. No. sc- 65334), phospho C-JUN (Santa cruz biotechnology, Inc. USA, Catelog. No. sc- 101723) and caspase 3 (Santa cruz biotechnology, Inc. USA, Catelog. No. sc-22171) was carried out in Panc-1 cells [referred to in step 1 of example 8] by using Cellomics high content imaging. Briefly, 1 X 10 4 cells were seeded in 96 well tissue culture grade black plate (Nunc brand, Thermofisher Scientific Inc. USA, Catalog. No. 165305) with transparent bottom and allowed them to adhere for 24 hrs.
  • step 2 of example 8 the media was replaced with fresh media and the cells were treated with 5 ⁇ , 3 ⁇ and 1 ⁇ concentrations of compound of formula (1) [referred to in step 2, of example 8] and incubated for 12 hrs and 24 hrs. After respective time point, cells were fixed with 3.7% formaldehyde (Sigma St. Louis, MO) for 10 minutes at RT, followed by permeabilization with 0.15% Triton X-100 (Sigma St. Louis, MO) for 10 minutes. Then the cells were blocked with 5% Bovine Serum Albumin for 2 hrs. After blocking step specific primary antibody were added for lhr and primary antibodies of different protein were localized by secondary antibody labeled with Dylight 548 (red) (Thermo Scientific, USA).
  • Dylight 548 red
  • Table 3 Effect of compound of formula (1) on p53, pC-JUN and cleaved caspase3 expression in Pane

Landscapes

  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Genetics & Genomics (AREA)
  • Biotechnology (AREA)
  • General Health & Medical Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biochemistry (AREA)
  • Microbiology (AREA)
  • General Engineering & Computer Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Virology (AREA)
  • Biomedical Technology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

La présente invention concerne un composé de formule (1). L'invention concerne l'intégralité des formes isomères et tautomères du composé de formule (1), ainsi que leurs sels et dérivés pharmaceutiquement acceptables, tels que leurs esters et éthers. La présente invention concerne, en outre, des procédés d'isolement et de production du composé de formule (1) impliquant la fermentation de la souche d'actinomycète marin PM0895117/MTCC 5675. L'invention concerne également les compositions pharmaceutiques contenant ledit composé de formule (1) en tant que principe actif et leur utilisation dans des médicaments destinés au traitement du cancer.
PCT/IB2013/052481 2012-03-30 2013-03-28 Analogue de nucléoside utilisable en tant que composé anticancéreux WO2013144894A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US201261617927P 2012-03-30 2012-03-30
US61/617,927 2012-03-30

Publications (1)

Publication Number Publication Date
WO2013144894A1 true WO2013144894A1 (fr) 2013-10-03

Family

ID=49258346

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/IB2013/052481 WO2013144894A1 (fr) 2012-03-30 2013-03-28 Analogue de nucléoside utilisable en tant que composé anticancéreux

Country Status (1)

Country Link
WO (1) WO2013144894A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2019044941A1 (fr) * 2017-09-01 2019-03-07 学校法人北里研究所 Nouveau composé présentant une activité thérapeutique contre une infection par le complexe mycobacterium avium et procédé pour sa production
CN109824746A (zh) * 2019-03-15 2019-05-31 杭州科兴生物化工有限公司 一种友霉素类化合物及其制备方法和应用

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
SHIOMI, K. ET AL.: "Cytosaminomycins, New Anicoccidial Agents Produced by Streptomyces sp. KO-8119. II. Structure Elucidation of Cytosaminomycins A, B, C and D", THE JOURNAL OF ANTIBIOTICS, vol. 47, no. 7, 1994, pages 782 - 786 *
STEVENS, C. L. ET AL.: "''Total Synthesis of the Amino Sugar Nucleoside Antibiotic, Plicacetin'' Journal of the American Chemical Society (1972), (94)93280-3281" *
ZHANG, G.: "Characterization of the Amicetin Biosynthesis Gene Cluster from Streptomyces vinaceusdrappus NRRL 2363 Implicates Two Alternative Strategies for Amide Bond Formation", APPLIED AND ENVIRONMENTAL MICROBIOLOGY, vol. 78, no. 7, 2012, pages 2393 - 2401 *

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2019044941A1 (fr) * 2017-09-01 2019-03-07 学校法人北里研究所 Nouveau composé présentant une activité thérapeutique contre une infection par le complexe mycobacterium avium et procédé pour sa production
CN111278985A (zh) * 2017-09-01 2020-06-12 学校法人北里研究所 具有针对鸟分枝杆菌复合体感染疾病治疗活性的新型化合物及其制造方法
JPWO2019044941A1 (ja) * 2017-09-01 2020-11-05 学校法人北里研究所 マイコバクテリウム・アビウム複合感染症に対する治療活性を有する新規化合物及びその製造方法
US11407742B2 (en) 2017-09-01 2022-08-09 The Kitasato Institute Compound having therapeutic activity against mycobacterium avium complex infection, and method for producing same
JP7184260B2 (ja) 2017-09-01 2022-12-06 学校法人北里研究所 マイコバクテリウム・アビウム複合感染症に対する治療活性を有する新規化合物及びその製造方法
CN111278985B (zh) * 2017-09-01 2023-09-08 学校法人北里研究所 具有针对鸟分枝杆菌复合体感染疾病治疗活性的新型化合物及其制造方法
CN109824746A (zh) * 2019-03-15 2019-05-31 杭州科兴生物化工有限公司 一种友霉素类化合物及其制备方法和应用

Similar Documents

Publication Publication Date Title
Hill Microbes from marine sponges: a treasure trove of biodiversity for natural products discovery
Giddings et al. Microbial natural products: molecular blueprints for antitumor drugs
Nikapitiya Bioactive secondary metabolites from marine microbes for drug discovery
Giddings et al. Bioactive compounds from marine extremophiles
WO2012010606A1 (fr) Polycétides hexacycliques en tant qu'inhibiteurs de kinases
Chen et al. Streptopyrazinones A− D, rare metabolites from marine-derived Streptomyces sp. ZZ446
US20160354430A1 (en) Hexadepsipeptide analogues as anticancer compounds
US20140348919A1 (en) Antibiotic compounds
WO2013144894A1 (fr) Analogue de nucléoside utilisable en tant que composé anticancéreux
WO2014106826A2 (fr) Analogue d'anthracycline et utilisations associées
JP5461082B2 (ja) ストレプトミセス属に属する新規微生物、その微生物が産生する新規化合物、及びその化合物を有効成分とする医薬
US20130130992A1 (en) Dipeptide derivative for the treatment of cancer
Cooreman et al. Emerging pharmaceutical therapies of Ascidian-derived natural products and derivatives
US20060167067A1 (en) Gm-95-containing antitumor effect potentiator, combined antitumor preparation and antitumor agent
JP5641716B2 (ja) ストレプトミセス属に属する微生物が産生する新規化合物、その化合物を産生する微生物、及びその化合物を有効成分とする医薬製剤
JP2012240974A (ja) ダイデムニンｂの製造方法
TW201444875A (zh) 六酯肽類似物作為抗癌化合物
TW201444864A (zh) 六酯肽類似物作為抗癌化合物
JP5578649B2 (ja) アクチノマデュラ属に属する新規微生物、その微生物が産生する新規化合物、及びその化合物を有効成分とする医薬
KR100225458B1 (ko) 믹소코크스 속의 점액세균, 이로부터 생산되는 신규 항종양활성 화합물 및 그의 제조방법
Figuerola Balañá et al. The Phylum Bryozoa as a promising source of anticancer drugs
EP0289354A2 (fr) L'antibiotique TAN-950A, sa production et son utilisation
JP2008074710A (ja) 新規ａ−９７０６５類物質
deMayo Localization and Investigation of Secondary Metabolite Producing Styela clava-Associated Streptomyces Bacteria
JP2010046004A (ja) ストレプトミセス属に属する新規微生物、その微生物が産生する新規化合物、及びその化合物を有効成分とする医薬

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 13768565

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 13768565

Country of ref document: EP

Kind code of ref document: A1