WO2019073296A1 - Formulations liposomales de bisantrène ou de dérivés ou analogues de celui-ci - Google Patents

Formulations liposomales de bisantrène ou de dérivés ou analogues de celui-ci Download PDF

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WO2019073296A1
WO2019073296A1 PCT/IB2018/001266 IB2018001266W WO2019073296A1 WO 2019073296 A1 WO2019073296 A1 WO 2019073296A1 IB 2018001266 W IB2018001266 W IB 2018001266W WO 2019073296 A1 WO2019073296 A1 WO 2019073296A1
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bisantrene
analog
group
composition
malignancy
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PCT/IB2018/001266
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John Rothman
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Race Oncology Ltd.
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/10—Dispersions; Emulsions
    • A61K9/127—Liposomes
    • 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
    • A61K31/4164—1,3-Diazoles
    • A61K31/4168—1,3-Diazoles having a nitrogen attached in position 2, e.g. clonidine
    • 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
    • A61K31/4164—1,3-Diazoles
    • A61K31/4178—1,3-Diazoles not condensed 1,3-diazoles and containing further heterocyclic rings, e.g. pilocarpine, nitrofurantoin
    • 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
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/513—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim having oxo groups directly attached to the heterocyclic ring, e.g. cytosine
    • 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
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
    • 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
    • A61K31/53—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with three nitrogens as the only ring hetero atoms, e.g. chlorazanil, melamine
    • 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
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
    • A61K31/7052—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides
    • A61K31/706—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom
    • A61K31/7064—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines
    • A61K31/7068—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines having oxo groups directly attached to the pyrimidine ring, e.g. cytidine, cytidylic 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
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
    • A61K31/7052—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides
    • A61K31/706—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom
    • A61K31/7064—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines
    • A61K31/7068—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines having oxo groups directly attached to the pyrimidine ring, e.g. cytidine, cytidylic acid
    • A61K31/7072—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines having oxo groups directly attached to the pyrimidine ring, e.g. cytidine, cytidylic acid having two oxo groups directly attached to the pyrimidine ring, e.g. uridine, uridylic acid, thymidine, zidovudine
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2300/00—Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00

Definitions

  • This invention is directed to liposomal formulations of bisantrene or derivatives or analogs thereof, particularly combination liposomal formulations of bisantrene together with cytarabine or another pyrimidine nucleoside analog.
  • biotherapeutics designed to stimulate the human immune system with vaccines, therapeutic antibodies, cytokines, lymphokines, inhibitors of tumor blood vessel development (angiogenesis) or gene and antisense therapies to alter the genetic make-up of cancer cells, and other biological response modifiers.
  • cancer is a collection of diseases with a multitude of etiologies and that a patient's response and survival from therapeutic intervention is complex with many factors playing a role in the success or failure of treatment including disease indication, stage of invasion and metastatic spread, patient gender, age, health conditions, previous therapies or other illnesses, genetic markers that can either promote or retard
  • Bisantrene generally employed as the dihydrochloride, is an unusual agent with direct cytotoxic action as well as genomic and immunologic methods of action.
  • the chemical name for bisantrene dihydrochloride is 9, 10- anthracenedicarboxaldehyde-bis [(4, 5-dihydro-1 H-imidazole-2-yl) hydrazine]
  • anthracycline chemotherapeutic agent drugs with planar structures based around a resonant aromatic ring structure that intercalates within the helices of DNA and disrupt various functions, including replication, presumably due to a strong inhibitory effect on the enzyme topoisomerase II. It was found that, like other anthracyclines, it could kill tumor cells in donogenic assays and intercalate with DNA, where it inhibits both DNA and RNA synthesis.
  • the primary chemotherapeutic mechanism for bisantrene is its preferential binding to A-T rich regions where it effects changes to supercoiling and initiates strand breaks in association with DNA associated proteins. This results from the inhibition of the enzyme topoisomerase II, which relaxes DNA coiling during replication. It was found that while inactive orally, intravenously (i.v.), intraperitoneally (i.p.), or
  • the drug was effective in cancer models using colon 26, Lewis lung, Ridgway osteosarcoma, B16, Lieberman plasma cell, P388 or L1210 cancer cells.
  • Activity in donogenic assays from 684 patients was seen in breast, small cell lung, large cell lung, squamous cell lung, ovarian, pancreatic, renal, adrenal, head and neck, sarcoma, gastric, lymphoma and melanoma tumor cells, but not in colorectal cancer. Importantly, a lack of cross resistance with Adriamycin and mitoxantrone was found.
  • bisantrene dihydrochloride has a number of toxicities. Toxicity studies in dogs and monkeys revealed that at high doses leukopenia, anorexia, diarrhea, injection site necrosis, enterocolitis, muscle degeneration, and pulmonary edema were observed. Although anthracyclines, have limited therapeutic utility due to their propensity to cause cardiac toxicity, the toxicity of bisantrene was observed to be less than that of any other agent in the anthracycline class.
  • Bisantrene is normally administered intravenously. However, the intravenous administration of bisantrene has been associated with severe local venous toxicity. Various alternatives have been tried to minimize this toxicity. In one
  • bisantrene doses have been infused via central venous access devices over 1 hour.
  • bisantrene has been infused through peripheral veins over 2 hours, and has been "piggybacked" into a running dextrose infusion in an attempt to lessen delayed swelling in the arm used for infusion.
  • patients have been given hydrocortisone (50 mg IV) and the antihistamine diphenhydramine (50 mg IM) immediately prior to bisantrene. Resultant nausea is frequently controlled with anti-emetic agents.
  • the nucleoside analog cytarabine has been used for the treatment of acute myeloid leukemia, acute lymphocytic leukemia, chronic myelogenous leukemia, and non-Hodgkin's lymphoma. It is an antimetabolite that can be incorporated into DNA in rapidly proliferating cells but then blocks DNA synthesis. It also inhibits both DNA and RNA polymerases and nucleotide reductases.
  • Liposomal formulations including a combination of the anthracycline derivative daunorubicin and cytarabine have been developed.
  • Daunorubicin is an intercalating agent that inhibits topoisomerase II.
  • Such formulations are disclosed in United States Patent No. 8,022,279 to Mayer et al.
  • One aspect of the present invention is a liposomal formulation including bisantrene or a derivative or analog thereof that is useful for achieving a drug retention and a sustained drug release for the bisantrene or the derivative or analog thereof.
  • the formulations are useful for treatment of neoplastic diseases and conditions including, but not limited to, acute myeloid leukemia, acute lymphocytic leukemia, and chronic myelogenous leukemia.
  • the composition comprises:
  • the composition comprises bisantrene.
  • the bisantrene is bisantrene dihydrochloride.
  • composition comprises a derivative or analog of bisantrene.
  • the derivative or analog of bisantrene can be selected from the group consisting of:
  • Ri and R3 are the same or different and are hydrogen, C1 -C6 alkyi, -C(0)-Rs, wherein R5 is hydrogen, C1 -C6 alkyi, phenyl, mono-substituted phenyl (wherein the substituent can be ortho, meta, or para and is fluoro, nitro, C1 -C6 alkyi, C1-C3 aikoxy, or cyano), pentafluorophenyl, naphthyl, furanyl,
  • Ri and R3 may be hydrogen or C1 -C6 alkyi
  • R2 and R 4 are the same or different and are: hydrogen, Ci -C 4 alkyi or -C(0)-R6, where R6 is hydrogen, C1 -C6 alkyi, phenyl, mono-substituted phenyl (wherein the substituent may be in the ortho, meta, or para position and is fluoro, nitro, C1 -C6 alkyi, C1-C3 aikoxy, or cyano), pentafluorophenyl, naphthyl, furanyl, or -ChbOChh; wherein the compounds can have the schematic structure B(Q)n, wherein B is the residue formed by removal of a hydrogen atom from one or more basic nitrogen atoms of an amine, amidine, guanidine, isourea, isothiourea, or biguanide-containing pharmaceutically active compound, and Q is hydrogen or A
  • R'O such that R' and R" are the same or different and are R (where R is C1-C6 alkyl, aryl, aralkyl, heteroalkyl, NC-CH2CH2-,
  • n is an integer representing the number of primary or secondary basic nitrogen atoms in the compound such that at least one Q is A;
  • the derivative or analog of bisantrene is a derivative of bisantrene selected from the group consisting of:
  • the composition comprises bisantrene, such as bisantrene dihydrochloride.
  • the liposomes comprise at least one lipid selected from the group consisting of a phosphatidylcholine lipid, a phosphatidylglycerol lipid, a sterol, an ether lipid, a phosphatidic acid, a phosphonate, a ceramide, a ceramide analog, a sphingosine, a sphingosine analog, a serine-containing lipid, a hydrophilic polymer-lipid conjugate, phosphatidylglycerol, phosphatidylinositol, and a negatively charged lipid having a hydrophilic portion and a hydrophobic portion with a neutral non-zwitterionic moiety attached to the hydrophilic portion of the lipid.
  • the liposomes comprise a phosphatidylcholine lipid such as diastearoylphosphatidylcholine.
  • the liposomes also comprise a phosphatidylglycerol lipid such as
  • the liposomes also comprise a sterol such as cholesterol.
  • the liposomes comprise diastearoylphosphatidylcholine, distearoylphosphatidylglycerol, and cholesterol.
  • the molar ratio of the distearoylphosphatidylcholine, the distearoylphosphatidylglycerol, and the cholesterol is from about 6.5 to about 7.5 of distearoylphosphatidylcholine, about 1.5 to about 2.5 of distearoylphosphatidylglycerol, and about 0.8 to 1 .2 of cholesterol.
  • the molar ratio of the distearoylphosphatidylcholine, the distearoylphosphatidylglycerol, and the cholesterol is from about 6.8 to about 7.2 of distearoylphosphatidylcholine, about 1 .8 to about 2.2 of distearoylphosphatidylglycerol, and about 0.9 to 1 .1 of cholesterol. More preferably, the molar ratio of the distearoylphosphatidylcholine, the
  • the liposomes have a diameter of less than about 300 nm. Preferably, the liposomes have a diameter of less than 200 nm. Typically, the liposomes have an intraliposomal osmolality of 500 mOSM/kg or less.
  • Another aspect of the present invention is a method for treating a malignancy comprising the step of administering a therapeutically effective quantity of a composition according to the present invention as described above to treat the malignancy.
  • the malignancy is selected from the group consisting of breast cancer, ovarian cancer, renal cancer, small-cell lung cancer, non-small cell lung cancer, Hodgkin's lymphoma, non-Hodgkin's lymphoma, acute myelocytic leukemia, melanoma, gastric cancer, adrenal cancer, head and neck cancer, hepatocellular cancer, hypernephroma, bladder cancer, acute leukemias of childhood, chronic lymphocytic leukemia, prostate cancer, glioblastoma, and myeloma.
  • the breast cancer can be selected from the group consisting of refractory breast cancer, triple-negative breast cancer, and breast cancer characterized by overexpressed Her-2-neu.
  • the malignancy is an acute leukemia of childhood
  • the acute leukemia of childhood can be selected from the group consisting of acute myelocytic leukemia (AML) and acute lymphocytic leukemia (ALL) of childhood.
  • the malignancy is prostate cancer
  • the prostate cancer can be androgen-resistant prostate cancer.
  • the malignancy is glioblastoma
  • the glioblastoma can be glioblastoma that is resistant to one or both of the following therapeutic agents:
  • the malignancy can be a malignancy that is characterized by overexpressed topoisomerase II or overexpressed and/or mutated EGFR.
  • Suitable liposomal compositions for use in methods according to the present invention are as described above.
  • the liposomal composition is administered parenterally, such as intravenously or intraperitoneally.
  • the liposomal composition is administered in a pharmaceutical formulation suitable for administration.
  • the method further comprises administration of a therapeutically effective quantity of an additional therapeutic agent.
  • the malignancy is breast cancer
  • the additional therapeutic agent can be selected from the group consisting of tamoxifen, anastrozole, letrozole, cyclophosphamide, docetaxel, paclitaxel, methotrexate, fluorouracil, and trastuzumab.
  • the malignancy is ovarian cancer
  • the additional therapeutic agent can be selected from the group consisting of: a platinum-containing antineoplastic drugs selected from the group consisting of cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatin,
  • phenanthriplatin picoplatin, and satraplatin
  • paclitaxel topotecan
  • gemcitabine
  • the malignancy is renal cancer
  • the additional therapeutic agent can be selected from the group consisting of
  • everolimus torisel, nexavar, sunitinib, axitinib, inferferon, interleukin-2, pazopanib, sorafenib, nivolumab, cabozanitib, and levanitib.
  • the sunitinib sunitinib
  • axitinib inferferon
  • interleukin-2 interleukin-2
  • pazopanib sorafenib
  • nivolumab cabozanitib
  • levanitib levanitib
  • the malignancy is small-cell lung cancer
  • the additional therapeutic agent can be selected from the group consisting of cyclophosphamide, cisplatin, etoposide, vincristine, paclitaxel, and carboplatin.
  • the malignancy is non- small-cell lung cancer
  • the additional therapeutic agent can be selected from the group consisting of cisplatin, erlotinib, gefitinib, afatinib, crizotinib, bevacizumab, carboplatin, paclitaxel, nivolumab, and pembrolizumab.
  • the malignancy is Hodgkin's lymphoma
  • the additional therapeutic agent can be selected from the group consisting of mechlorethamine, vincristine, prednisone, procarbazine, bleomycin, vinblastine, dacarbazine, etoposide, and cyclophosphamide.
  • the malignancy is non-Hodgkin's lymphoma
  • the additional therapeutic agent can be selected from the group consisting of cyclophosphamide, vincristine, and prednisone.
  • the malignancy is acute myelocytic leukemia
  • the additional therapeutic agent can be selected from the group consisting of cytarabine, fludarabine, all-frans-retinoic acid, interleukin-2, and arsenic trioxide.
  • the malignancy is melanoma
  • the additional therapeutic agent can be selected from the group consisting of temozolomide, dacarbazine, interferon, interleukin-2, ipilimumab, pembrolizumab, nivolumab, vemurafenib, dabrafenib, and trametinib.
  • the malignancy is gastric cancer, and the additional therapeutic agent can be selected from the group consisting of 5-fluorouracil, capecitabine, carmustine, semustine, mitomycin C, cisplatin, taxotere, and trastuzumab.
  • the malignancy is adrenal cancer, and the additional therapeutic agent can be selected from the group consisting of mitotane, cisplatin, etoposide, and streptozotocin.
  • the malignancy is head and neck cancer, and the additional therapeutic agent can be selected from the group consisting of paclitaxel, carboplatin, cetuximab, docetaxel, cisplatin, and 5-fluorouracil.
  • the malignancy is hepatocellular cancer
  • the additional therapeutic agent can be selected from the group consisting of tamoxifen, octreoside, synthetic retinoids, cisplatin, 5-fluorouracil, interferon, taxol, and sorafenib.
  • the malignancy is hypernephroma
  • the additional therapeutic agent can be selected from the group consisting of nivolumab, everolimus, sorafenib, axitinib, lenvatinib, temsirolimus, sunitinib, pazopanib, interleukin- 2, cabozanitib, bevacizumab, interferon a, ipilimumab, atezolizumab, varilumab, durvalumab, tremelimumab, and avelumab.
  • the malignancy is bladder cancer
  • the additional therapeutic agent can be selected from the group consisting of cisplatin, 5-fluorouracil, mitomycin C, gemcitabine, methotrexate, vinblastine, carboplatin, paclitaxel, docetaxel, ifosfamide, and pemetrexed.
  • the malignancy is acute myelocytic leukemia of childhood
  • the additional therapeutic agent can be selected from the group consisting of methotrexate, nelarabine, asparaginase, blinatumomab, cyclophosphamide, clofarabine, cytarabine, dasatinib, methotrexate, imatinib, pomatinib, vincristine, 6-mercaptopurine,
  • the malignancy is acute lymphocytic leukemia
  • the additional therapeutic agent can be selected from the group consisting of asparaginase, vincristine, dexamethasone, methotrexate, 6- mercaptopurine, cytarabine, hydrocortisone, 6-thioguanine, prednisone, etoposide, cyclophosphamide, mitoxantrone, and teniposide.
  • the malignancy is chronic lymphocytic leukemia
  • the additional therapeutic agent can be selected from the group consisting of fludarabine, cyclophosphamide, rituximab, vincristine, prednisolone, bendamustine, alemtuzumab, ofatumumab, obinutuzumab, ibrutinib, idelalisib, and venetoclax.
  • the malignancy is prostate cancer
  • the additional therapeutic agent can be selected from the group consisting of temozolomide, docetaxel, cabazitaxel, bevacizumab, thalidomide, prednisone, sipuleucel-T, abiraterone, and enzalutamide.
  • the malignancy is glioblastoma
  • the additional therapeutic agent can be selected from the group consisting of temozolomide and bevacizumab.
  • the malignancy is myeloma and the additional therapeutic agent can be selected from the group consisting of bortezomib, lenalidomide, dexamethasone, melphalan, prednisone, thalidomide, and cyclophosphamide.
  • Another aspect of the present invention is a liposomal formulation including both bisantrene or a derivative or analog thereof and a nucleoside analog such as cytarabine that is useful for achieving a drug retention and a sustained drug release for each of the two therapeutic agents.
  • the formulations are useful for treatment of neoplastic diseases and conditions including, but not limited to, acute myeloid leukemia, acute lymphocytic leukemia, and chronic myelogenous leukemia.
  • the composition comprises:
  • the pyrimidine analog antimetabolite is typically selected from the group consisting of cytarabine, 5-azacytidine, gemcitabine, floxuridine, 5-fluorouracil, capecitabine, 6-azauracil, troxacitabine, thiarabine, sapacitabine, CNDAC (2'-cyano-2'- deoxy-1 - ⁇ -D-arabinofuranosylcytosine), 2'-deoxy-2'-methylidenecytidine, 2'-deoxy-2'- fluoromethylidenecytidine, 2'-deoxy-2'-methylidene-5-fluorocytidine, 2'-deoxy-2',2'- difluorocytidine, and 2'-C-cyano-2'-deoxy- -arabinofuranosylcytosine.
  • the pyrimidine analog antimetabolite is selected from the group consisting of cytarabine, 5- azacytidine, gemcitabine, floxuridine, 5-fluorouracil, capecitabine, and 6-azauracil. More preferably, the pyrimidine analog antimetabolite is cytarabine.
  • Bisantrene derivatives and analogs suitable for use in the composition are as described above.
  • the bisantrene or derivative or analog thereof is bisantrene, such as bisantrene dihydrochloride.
  • a preferred composition according to the present invention comprises bisantrene and cytarabine.
  • the ratio of the pyrimidine analog antimetabolite and the bisantrene or derivative or analog thereof is from about 25: 1 to about 1 : 1.
  • the ratio of the pyrimidine analog antimetabolite and the bisantrene or derivative or analog thereof is from about 10: 1 to about 3: 1. More preferably, the ratio of the pyrimidine analog antimetabolite and the bisantrene or derivative or analog thereof is about 5:1 .
  • compositions according to the present invention can be used in a method for treating a malignancy as described above. Suitable methods and malignancies to be treated are as described above. Additional therapeutic agents as described above can also be used, with the proviso that a pyrimidine analog antimetabolite used in the composition is not generally used as an additional therapeutic agent.
  • One aspect of the invention is a composition of matter comprising: (i) a therapeutically effective quantity of bisantrene or a derivative or analog thereof; and (ii) a liposome encapsulating both the therapeutically effective quantity of bisantrene or the derivative or analog thereof.
  • Bisantrene is a tricyclic aromatic compound with the chemical name, 9, 10-anthracenedicarboxaldehyde bis[(4,5-dihydro-1 H-imidazol-2-yl)hydrazine] dihydrochloride.
  • the molecular formula is C22H22N8 ⁇ 2HCI and the molecular weight, 471 .4.
  • the alkylimidazole side chains are very basic and, at physiologic pH, are positively charged. This is believed to facilitate electrostatic attractions to negatively charged ribose phosphate groups in DNA.
  • Human tumor cells that were sensitive to bisantrene as assessed by in vitro colony-forming assays include breast cancer, ovarian cancer, renal cancer, small cell and non-small cell lung cancer, lymphoma, acute myelogenous leukemia, melanoma, gastric cancer, adrenal cancer, and head and neck cancer (D.D.
  • bisantrene vials have been reconstituted with 2 to 5 ml_ of Sterile Water for Injection, USP, and then diluted with approximately 0.1 to 0.5 mg/ml_ in D5W (5% dextrose in water).
  • Bisantrene is incompatible with saline and unstable in light (G. Powis et al., "Pharmacokinetic Study of ADAH in Humans and Sensitivity of ADAH to Light" (Abstract #C-74)," ASCO Proc. 1 : 19 (1982).
  • the drug may be metabolized to some extent in vivo.
  • In vitro bisantrene is a substrate for hepatic microsomal enzymes but specific metabolites have not been identified.
  • Preclinical drug distribution studies showed that the tissues with the highest concentration (in descending order) are kidney, liver, gallbladder, spleen, lung, and heart. Brain levels were extremely low. The drug did distribute to lymph nodes and bone marrow (W.H. Wu & G. Nicolau, "Disposition and Metabolic Profile of a New Antitumor Agent, CL 216,942 (Bisantrene) in Laboratory Animals," Cancer Treat Rep. 66: 1 173-1 185 (1982)).
  • Desai et al. discloses a composition of matter for delivery of a hydrophobic drug (i.e., bisantrene or a derivative or analog thereof) comprising: (i) the hydrophobic drug; (ii) an oleaginous vehicle or oil phase that is substantially free of butylated hydroxyanisole (BHA) or butylated hydroxytoluene (BHT); (iii) a co-surfactant or emulsifier; (iv) a co-surfactant or auxiliary emulsifier; and (v) benzyl alcohol as a co- solvent.
  • a hydrophobic drug i.e., bisantrene or a derivative or analog thereof
  • a pharmaceutically acceptable oleaginous vehicle or oil selected from the group consisting of: (a) naturally occurring vegetable oils and (b) semisynthetic mono-, di-, and triglycerides, wherein the oleaginous vehicle or oil is free of BHT or BHA; (iii) a surfactant or emulsifier; (iv) a co-surfactant or emulsifier; (v) an ion-pair former selected from C6-C20 saturated or unsaturated aliphatic acids when the group consisting of: (a) naturally occurring vegetable oils and (b) semisynthetic mono-, di-, and triglycerides, wherein the oleaginous vehicle or oil is free of BHT or BHA; (iii) a surfactant or emulsifier; (iv) a co-surfactant or emulsifier; (v) an ion-pair former selected from C6-C20 saturated or unsaturated aliphatic acids
  • hydrophobic drug is basic and a pharmaceutically acceptable aromatic amine when the hydrophobic drug is acidic; and (vi) water.
  • United States Patent No. 5,000,886 to Lawter et al. and United States Patent No. 5, 143,661 to Lawter et al. disclose
  • compositions for delivery of pharmaceutical agents such as bisantrene or a derivative or analog thereof comprising a microcapsule, wherein the microcapsule includes a hardening agent that is a volatile silicone fluid.
  • United States Patent No. 5,070,082 to Murdock et al., United States Patent No. 5,077,282 to Murdock et al., and United States Patent No. 5,077,283 to Murdock et al. disclose prodrug forms of poorly soluble hydrophobic drugs, including bisantrene and derivatives and analogs, that are salts of a phosphoramidic acid.
  • United States Patent No. 5,378,456 to Tsou includes compositions containing an anthracene antitumor agent, such as bisantrene or a derivative or analog thereof, in which the bisantrene or derivative or analog thereof is conjugated to or admixed with a divinyl ether-maleic acid (MVE) copolymer.
  • MVE divinyl ether-maleic acid
  • United States Patent No. 5,609,867 to Tsou discloses polymeric 1 ,4-bis derivatives of bisantrene and copolymers of bisantrene and another monomer, such as a dianhydride.
  • the present application therefore, provides improved formulations for the use of bisantrene and analogs or derivatives thereof together with nucleoside analogs for the treatment of malignancies while avoiding the side effects described above and improving the therapeutic efficacy of the drug.
  • the present application also provides methods for administration of formulations according to the present invention for the treatment of malignancies and other diseases and conditions as described below.
  • bisantrene in addition to direct antineoplastic effects related to the activity of bisantrene as a DNA intercalator, bisantrene also possesses other
  • NKG2D is an activating receptor that is involved in tumor immunosurveillance by NK cells, NKT cells, ⁇ T cells and resting (in mice) and/or activated (in humans) CD8 + T cells, and also states that anthracycline-based agents may act as immunostimulators, particularly in combination with IL-12; such agents also promote HMGB1 release and activate T cells;
  • TLR2 and TLR9 are Sensors of Apoptosis in a Mouse Model of Doxorubicin-lnduced Acute Inflammation," Cell Death Different. 18: 1316-1325 (201 1 ), which states that anthracycline-based antibiotics induce an immunogenic form of apoptosis that has immunostimulatory properties mediated by MyD88, TLR2, and TLR9;
  • bladder carcinoma multiple myeloma, lung adenocarcinoma, melanoma, and renal cell carcinoma
  • breast cancer Boden et al. (1985), supra
  • acute myelogenous leukemia especially relapsed or refractory acute myeloid leukemia
  • A. Spadea et al. "Bisantrene in Relapsed and Refractory Myelogenous Leukemia,” Leukemia Lymphoma 9: 217-220 (1993)
  • a number of macrophage activators are known, including Bacillus Calmette-Guerin, Corynebacterium parvum, endotoxins, muramyl dipeptide, pl:pC copolymer, pyran copolymer, lymphokines, Adriamycin, cyclophosphamide, and mitomycin C.
  • the efficacy of bisantrene in allogeneic macrophage transplants and with supernatants of macrophages activated by bisantrene has been shown in B.S. Wang et al., "Immunotherapy of a Murine
  • Telomerase is a ribonucleoprotein reverse transcriptase responsible for maintenance of telomere length. Its expression is associated with cell immortalization and tumorigenesis since it is expressed in most human tumor cells but is not active in most normal somatic cells. Telomerase machinery inhibitors have been evaluated as potential anticancer agents, including nucleotide analogs such as 7-deaza-2'- deoxyguanosine, BIBR1532 (2-[[(E)-3-naphthalen-2-ylbut-2-enoyl]amino]benzoic acid), antisense oligonucleotides, imetelstat sodium, and other agents. For such agents, a number of different pathways are involved in inhibition of telomerase activity.
  • telomere inhibition results in cellular senescence or apoptosis in a time-dependent manner that correlates with the initial telomere length in the cells in which telomerase is inhibited.
  • telomere architecture collapses or is disrupted a signaling cascade comparable to that produced by DNA damage is activated and cell cycle arrest (accelerated senescence) or cell death through apoptosis is induced.
  • Telomerase substrates are the telomeres, double-stranded DNA portions with a 3' protruding overhang (100-200 bases long), formed by a repeating noncoding sequence (TTAGGG (SEQ ID NO: 1 ) in humans).
  • TTAGGG SEQ ID NO: 1
  • the single-stranded portion can fold into a structure called G-quadruplex.
  • G-quadruplex a structure called G-quadruplex.
  • Hoogsteen-paired guanines are between the N7 position of the purine base as a hydrogen-bond acceptor and the C6 amino group of the pyrimidine base as a donor.
  • telomere binding proteins i.e., TRF2 and hPOT1
  • TRF2 and hPOT1 telomere binding proteins
  • Several compounds able to interact with and stabilize G-quadruplex structures formed by G-rich single-stranded overhangs of telomeres have been identified, including anthraquinones, fluorenones, acridines, triazine, cationic porphyrins, and perylenes, as well as other compounds. These compounds share a general consensus structural motif based on a large flat aromatic surface linked to protonatable side chains. DNA binding occurs mainly through stacking on a terminal G-tetrad, whereas side chains contribute to the stability of the complex by hydrophobic/ionic interactions into the DNA grooves.
  • G- quadruplex binders such as the tri-substituted acridine BRAC019 (N,N'-(9- ⁇ [4- (dimethylamino)phenyl]amino ⁇ acridine-3,6-diyl)bis(3-pyrrolidin-1 -ylpropanamide) trihydroch!oride) and the 2,6 or 2,7 bis-substituted amido-anthraquinones.
  • binders are characterized by poor cytotoxicity and are able to induce a reduction in telomere length upon long-term drug exposure.
  • Bisantrene shares the structural "consensus motif" characteristic of effective G-quadruplex binders.
  • At least two side chains with amine groups protonatable at physiological pH are required for G-quadruplex binding.
  • Bisantrene is believed to intercalate between adjacent base pairs of double-stranded DNA through ⁇ - ⁇ stacking, with side chains located in either groove (threading mode), which grants affinity constants well above 10 6 M "1 under physiological conditions.
  • the fact that the most efficient G-quadruplex binders are substituted on two distinct aromatic rings with side chains pointing in opposite directions with reference to the long axis of the aromatic system likely suggests formation of additional specific interactions between the 4,5-dihydro-1 H-imidazol-2-yl hydrazone groups and the G-quadruplex structure.
  • At least one of the bisantrene analogs, Formula (III), has the ability to act both at the telomerase level, by interfering with substrate recognition (hence
  • telomere function not only in telomerase- expressing cells but also in ALT-positive cell lines, since it consistently provokes a DNA damage response, as evidenced by the formation of ⁇ 2 ⁇ foci that partially co-localize at the telomere, in agreement with results reported for telomestatin.
  • a DNA damage response together with the absence of apoptosis and the induction of cell cycle impairment (mainly G2M phase arrest), suggest a drug-mediated activation of a senescence pathway.
  • HL-37 is anthracen-9-ylmethylene-[2-methoxyethoxymethylsulfanyl]-5-pyridin-3-yl-[1 ,2,4]triazol-4- amine and has the structure shown below as Formula (IX):
  • Bisantrene-amsacrine hybrids are also disclosed in G. Capranico et al., “Mapping Drug Interactions at the Covalent Topoisomerase ll-DNA Complex by Bisantrene/Amsacrine Congeners," J. Biol. Chem. 273: 12732-12739 (1998). These compounds are depicted below as Formulas (X), (XI), (XII), and (XIII): [0065] Additional derivatives and analogs of bisantrene include the
  • Ri and R3 are the same or different and are hydrogen, C1-C6 alkyl, -C(0)-Rs, wherein R5 is hydrogen, C1-C6 alkyl, phenyl, mono-substituted phenyl (wherein the substituent can be ortho, meta, or para and is fiuoro, nitro, C1-C6 alkyl, C1-C3 alkoxy, or cyano), pentafluorophenyl, naphthyl, furanyl,
  • Ri and R3 may be hydrogen or C1-C6 alkyl
  • R2 and R 4 are the same or different and are: hydrogen, Ci-C 4 alkyl or -C(0)-R6, where R6 is hydrogen, C1-C6 alkyl, phenyl, mono-substituted phenyl (wherein the substituent may be in the ortho, meta, or para position and is fluoro, nitro, C1-C6 alkyl, C1-C3 alkoxy, or cyano), pentafluorophenyl, naphthyl, furanyl, or -ChbOChh.
  • the compounds can have the schematic structure B(Q)n, wherein B is the residue formed by removal of a hydrogen atom from one or more basic nitrogen atoms of an amine, amidine, guanidine, isourea, isothiourea, or biguanide-containing pharmaceutically active compound, and Q is hydrogen or A, wherein A is
  • R"0 such that R' and R" are the same or different and are R (where R is C1-C6 alkyl, aryl, aralkyl, heteroalkyl, NC-CH2CH2-,
  • n is an integer representing the number of primary or secondary basic nitrogen atoms in the compound such that at least one Q is A.
  • These compounds include 9, 10-bis[(2-hydroxyethyl)iminomethyl]anthracene; 9, 10-bis ⁇ [2-(-2- hydroxyethylamino)ethyl]iminomethyl ⁇ anthracene; 9, 10-bis ⁇ [2-(morpholin-4- yl)ethyl]iminomethyl ⁇ anthracene; 9, 10-bis[(2-hydroxyethyl)aminomethyl]anthracene; 9, 10-bis ⁇ [2-(2-hydroxyethylamino)ethyl]aminomethyl ⁇ anthracene tetrahydrochloride; 9, 10-bis ⁇ [2-(piperazin-1 -yl)ethyl]aminomethyl ⁇ anthracene hexahydrochloride; and 9, 10- bis ⁇ [2-(morpholin-4-yl)ethyl]aminomethyl ⁇ anthracene tetrahydrochloride.
  • the term "derivative" as applied to bisantrene refers to a compound that has the same carbon skeleton as bisantrene, including the tricyclic aromatic nucleus and the two side chains attached to the tricyclic aromatic nucleus but has one or more substituents as described below that replace at least one hydrogen present in bisantrene with another moiety.
  • the term “analog” as applied to bisantrene applies to a compound related structurally to bisantrene but alters one or more of the tricyclic aromatic nucleus or one or more of the side chains, for example, by replacing one or more carbons in the tricyclic aromatic nucleus with nitrogens or by removing or moving one or both of the side chains.
  • Analogs of bisantrene include, but are not limited to compounds described above as Formulas (ll)-(XIV), as well as additional compounds described above and their derivatives.
  • derivatives and analogs of bisantrene can be optionally substituted with one or more groups that do not substantially affect the pharmacological activity of the derivative or analog.
  • groups are generally known in the art. Definitions for a number of common groups that can be used as optional substituents are provided below; however, the omission of any group from these definitions cannot be taken to mean that such a group cannot be used as an optional substituent as long as the chemical and pharmacological requirements for an optional substituent are satisfied.
  • alkyl refers to an unbranched, branched, or cyclic saturated hydrocarbyl residue, or a combination thereof, of from 1 to 12 carbon atoms that can be optionally substituted; the alkyl residues contain only C and H when unsubstituted.
  • the unbranched or branched saturated hydrocarbyl residue is from 1 to 6 carbon atoms, which is referred to herein as "lower alkyl.”
  • the hydrocarbyl residue includes at least three carbon atoms, which is the minimum number to form a ring.
  • alkenyl refers to an unbranched, branched or cyclic hydrocarbyl residue having one or more carbon-carbon double bonds.
  • alkynyl refers to an unbranched, branched, or cyclic hydrocarbyl residue having one or more carbon-carbon triple bonds; the residue can also include one or more double bonds. With respect to the use of “alkenyl” or “alkynyl,” the presence of multiple double bonds cannot produce an aromatic ring.
  • hydroxyalkyl refers to an alkyl, alkenyl, or alkynyl group including one or more hydroxyl groups as substituents; as detailed below, further substituents can be optionally included.
  • aryl refers to a monocyclic or fused bicyclic moiety having the well-known characteristics of aromaticity; examples include phenyl and naphthyl, which can be optionally substituted.
  • hydroxyaryl refers to an aryl group including one or more hydroxyl groups as substituents; as further detailed below, further substituents can be optionally included.
  • heteroaryl refers to monocyclic or fused bicylic ring systems that have the characteristics of aromaticity and include one or more
  • heteroatoms selected from 0, S, and N.
  • the inclusion of a heteroatom permits aromaticity in 5-membered rings as well as in 6-membered rings.
  • heteroaromatic systems include monocyclic C5-C6 heteroaromatic groups such as pyridyl, pyrimidyl, pyrazinyl, thienyl, furanyl, pyrrolyl, pyrazolyl, thiazolyl, oxazolyl, triazolyl, triazinyl, tetrazolyl, tetrazinyl, and imidazolyl, as well as the fused bicyclic moieties formed by fusing one of these monocyclic heteroaromatic groups with a phenyl ring or with any of the heteroaromatic monocyclic groups to form a Cs-C-io bicyclic group such as indolyl, benzimidazolyl, indazolyl, benzotriazolyl, isoquinolyl, quinolyl, benzothiazolyl, benzofuranyl, pyrazolylpyridyl, quinazolinyl, quinoxalinyl
  • any monocyclic or fused ring bicyclic system that has the characteristics of aromaticity in terms of delocalized electron distribution throughout the ring system is included in this definition.
  • This definition also includes bicyclic groups where at least the ring that is directly attached to the remainder of the molecule has the characteristics of aromaticity, including the delocalized electron distribution that is characteristic of aromaticity.
  • the ring systems contain 5 to 12 ring member atoms and up to four heteroatoms, wherein the heteroatoms are selected from the group consisting of N, 0, and S.
  • the monocyclic heteroaryls contain 5 to 6 ring members and up to three heteroatoms selected from the group consisting of N, 0, and S; frequently, the bicyclic heteroaryls contain 8 to 10 ring members and up to four heteroatoms selected from the group consisting of N, 0, and S.
  • the number and placement of heteroatoms in heteroaryl ring structures is in
  • hydroxheteroaryl refers to a heteroaryl group including one or more hydroxyl groups as substituents; as further detailed below, further substituents can be optionally included.
  • haloaryl and haloheteroaryl refer to aryl and heteroaryl groups, respectively, substituted with at least one halo group, where "halo” refers to a halogen selected from the group consisting of fluorine, chlorine, bromine, and iodine, typically, the halogen is selected from the group consisting of chlorine, bromine, and iodine; as detailed below, further substituents can be optionally included.
  • haloalkyl refers to alkyl, alkenyl, and alkynyl groups, respectively, substituted with at least one halo group
  • halo refers to a halogen selected from the group consisting of fluorine, chlorine, bromine, and iodine, typically, the halogen is selected from the group consisting of chlorine, bromine, and iodine; as detailed below, further substituents can be optionally included.
  • optionally substituted indicates that the particular group or groups referred to as optionally substituted may have no non- hydrogen substituents, or the group or groups may have one or more non-hydrogen substituents consistent with the chemistry and pharmacological activity of the resulting molecule. If not otherwise specified, the total number of such substituents that may be present is equal to the total number of hydrogen atoms present on the unsubstituted form of the group being described; fewer than the maximum number of such
  • substituted whether used as part of "optionally substituted” or otherwise, when used to modify a specific group, moiety, or radical, means that one or more hydrogen atoms are, each, independently of each other, replaced with the same or different substituent or substituents.
  • NZ b C(O)NZ c Z c — NZ b C(NZ b )Z b ,— NZ b C(NZ b )NZ c Z c , wherein Z a is selected from the group consisting of alkyl, cycloalkyl, heteroalkyl, cycloheteroalkyl, aryl, arylalkyl, heteroaryl and heteroarylalkyl; each Z b is independently hydrogen or Z a ; and each Z c is independently Z b or, alternatively, the two Z c 's may be taken together with the nitrogen atom to which they are bonded to form a 4-, 5-, 6-, or 7-membered cycloheteroalkyl ring structure which may optionally include from 1 to 4 of the same or different heteroatoms selected from the group consisting of N, O, and S.
  • — NZ C Z C is meant to include— NH2,— NH-alkyl,— N-pyrrolidinyl, and— N-morpholinyl, but is not limited to those specific alternatives and includes other alternatives known in the art.
  • a substituted alkyl is meant to include— alkylene-O-alkyl,— alkylene-heteroaryl,— alkylene-cycloheteroaryl,— alkylene- C(O)OZ b ,— alkylene-C(O)NZ b Z b , and— CH ⁇ CH 2 — C(O)-CH 3 , but is not limited to those specific alternatives and includes other alternatives known in the art.
  • the one or more substituent groups, together with the atoms to which they are bonded, may form a cyclic ring, including, but not limited to, cycloalkyl and cycloheteroalkyl.
  • substituent groups useful for substituting unsaturated carbon atoms in the specified group, moiety, or radical include, but are not limited to,— Z a , halo,— O-,— OZ b ,— SZ b ,— S " ,— NZ C Z C , trihalomethyl,— CF 3 ,— CN,— OCN,— SCN, —NO,— NO 2 ,— N 3 ,— S(O) 2 Z b ,— S(O 2 )O-,— S(O 2 )OZ b ,— OS(O 2 )OZ b ,— OS(O 2 )O " ,— P(O)(O-) 2 ,— P(O)(OZ b )(O " ),— P(O)(OZ b )(OZ b ),— C(O)Z b ,— C(S)Z b ,— C(NZ b )Z b
  • substituent groups useful for substituting nitrogen atoms in heteroalkyl and cycloheteroalkyl groups include, but are not limited to,— Z a , halo,— O " , — OZ b ,— SZ b ,— S " ,— NZ C Z C , trihalomethyl,— CF 3 ,— CN,—OCN,—SCN,—NO,— NO 2 ,— S(O) 2 Z b ,— S(O 2 )O " ,— S(O 2 )OZ b ,— OS(O 2 )OZ b ,— OS(O 2 )O " ,— P(O)(O ) 2 ,— P(0)(OZ b )(0-),— P(0)(OZ b )(OZ b ),— C(0)Z b ,— C(S)Z b ,— C(NZ b )Z b ,— C(0)OZ
  • the compounds described herein may contain one or more chiral centers and/or double bonds and therefore, may exist as stereoisomers, such as double-bond isomers (i.e., geometric isomers such as E and Z), enantiomers or diastereomers.
  • stereoisomers such as double-bond isomers (i.e., geometric isomers such as E and Z), enantiomers or diastereomers.
  • the invention includes each of the isolated stereoisomeric forms (such as the
  • stereoisomerically pure form e.g., geometrically pure, enantiomerically pure or diastereomerically pure
  • enantiomeric and stereoisomeric mixtures can be resolved into their component enantiomers or stereoisomers using separation techniques or chiral synthesis techniques well known to the skilled artisan.
  • the invention includes each of the isolated stereoisomeric forms as well as mixtures of stereoisomers in varying degrees of chiral purity, including racemic mixtures. It also encompasses the various diastereomers. Other structures may appear to depict a specific isomer, but that is merely for convenience, and is not intended to limit the invention to the depicted isomer. When the chemical name does not specify the isomeric form of the compound, it denotes any one of the possible isomeric forms or mixtures of those isomeric forms of the compound.
  • the compounds may also exist in several tautomeric forms, and the depiction herein of one tautomer is for convenience only, and is also understood to encompass other tautomers of the form shown. Accordingly, the chemical structures depicted herein encompass all possible tautomeric forms of the illustrated compounds.
  • tautomer refers to isomers that change into one another with great ease so that they can exist together in equilibrium. For example, ketone and enol are two tautomeric forms of one compound.
  • solvate means a compound formed by solvation (the combination of solvent molecules with molecules or ions of the solute), or an aggregate that consists of a solute ion or molecule, i.e., a compound of the invention, with one or more solvent molecules.
  • solvate is a “hydrate.” Examples of hydrates include, but are not limited to,
  • pharmaceutically acceptable salt and/or prodrug of the present compound may also exist in a solvate form.
  • the solvate is typically formed via hydration which is either part of the preparation of the present compound or through natural absorption of moisture by the anhydrous compound of the present invention.
  • ester means any ester of a present compound in which any of the -COOH functions of the molecule is replaced by a -COOR function, in which the R moiety of the ester is any carbon-containing group which forms a stable ester moiety, including but not limited to alkyl, alkenyl, alkynyl, cycloalkyl,
  • hydrolyzable esters of the present compounds are the compounds whose carboxyls are present in the form of hydrolyzable ester groups. That is, these esters are pharmaceutically acceptable and can be hydrolyzed to the corresponding carboxyl acid in vivo.
  • alkyl, alkenyl and alkynyl groups can alternatively or in addition be substituted by C-i-Cs acyl, C2-C8 heteroacyl, C6-C10 aryl, C3-C8 cycloalkyl, C3-C8 heterocyclyl, or C5-C10 heteroaryl, each of which can be optionally substituted.
  • the two groups capable of forming a ring having 5 to 8 ring members are present on the same or adjacent atoms, the two groups can optionally be taken together with the atom or atoms in the substituent groups to which they are attached to form such a ring.
  • Heteroalkyl “heteroalkenyl,” and “heteroalkynyl” and the like are defined similarly to the corresponding hydrocarbyl (alkyl, alkenyl and alkynyl) groups, but the 'hetero' terms refer to groups that contain 1 -3 0, S or N heteroatoms or combinations thereof within the backbone residue; thus at least one carbon atom of a corresponding alkyl, alkenyl, or alkynyl group is replaced by one of the specified heteroatoms to form, respectively, a heteroalkyl, heteroalkenyl, or heteroalkynyl group.
  • such groups do not include more than two contiguous heteroatoms except where an oxo group is present on N or S as in a nitro or sulfonyl group.
  • alkyl as used herein includes cycloalkyl and cycloalkylalkyl groups
  • cycloalkyl may be used herein to describe a carbocyclic non-aromatic group that is connected via a ring carbon atom
  • cycloalkylalkyl may be used to describe a carbocyclic non-aromatic group that is connected to the molecule through an alkyl linker.
  • heterocyclyl may be used to describe a non-aromatic cyclic group that contains at least one heteroatom (typically selected from N, 0 and S) as a ring member and that is connected to the molecule via a ring atom, which may be C (carbon-linked) or N (nitrogen-linked); and “heterocyclylalkyl” may be used to describe such a group that is connected to another molecule through a linker.
  • the heterocyclyl can be fully saturated or partially saturated, but non-aromatic.
  • substituents that are suitable for the cycloalkyl, cycloalkylalkyl, heterocyclyl, and heterocyclylalkyl groups are the same as those described above for alkyl groups.
  • the heterocyclyl groups typically contain 1 , 2 or 3 heteroatoms, selected from N, 0 and S as ring members; and the N or S can be substituted with the groups commonly found on these atoms in heterocyclic systems. As used herein, these terms also include rings that contain a double bond or two double bonds, as long as the ring that is attached is not aromatic.
  • the substituted cycloalkyl and heterocyclyl groups also include cycloalkyl or heterocyclic rings fused to an aromatic ring or heteroaromatic ring, provided the point of attachment of the group is to the cycloalkyl or heterocyclyl ring rather than to the aromatic/heteroaromatic ring.
  • acyl encompasses groups comprising an alkyl, alkenyl, alkynyl, aryl or arylalkyi radical attached at one of the two available valence positions of a carbonyl carbon atom
  • heteroacyl refers to the corresponding groups wherein at least one carbon other than the carbonyl carbon has been replaced by a heteroatom chosen from N, 0 and S.
  • Acyl and heteroacyl groups are bonded to any group or molecule to which they are attached through the open valence of the carbonyl carbon atom.
  • C-i-Cs acyl groups which include formyl, acetyl, pivaloyl, and benzoyl
  • C2-C8 heteroacyl groups which include methoxyacetyl, ethoxycarbonyl, and 4-pyridinoyl.
  • arylalkyi and “heteroarylalkyl” refer to aromatic and
  • heteroaromatic ring systems which are bonded to their attachment point through a linking group such as an alkylene, including substituted or unsubstituted, saturated or unsaturated, cyclic or acyclic linkers.
  • linker is C-i-Cs alkyl.
  • These linkers may also include a carbonyl group, thus making them able to provide substituents as an acyl or heteroacyl moiety.
  • An aryl or heteroaryl ring in an arylalkyi or heteroarylalkyl group may be substituted with the same substituents described above for aryl groups.
  • an arylalkyi group includes a phenyl ring optionally substituted with the groups defined above for aryl groups and a C1-C4 alkylene that is unsubstituted or is substituted with one or two C1-C4 alkyl groups or heteroalkyl groups, where the alkyl or heteroalkyl groups can optionally cyclize to form a ring such as cyclopropane, dioxolane, or oxacyclopentane.
  • a heteroarylalkyl group preferably includes a C5-C6 monocyclic heteroaryl group that is optionally substituted with the groups described above as substituents typical on aryl groups and a C1-C4 alkylene that is unsubstituted or is substituted with one or two C1-C4 alkyl groups or heteroalkyl groups, or it includes an optionally substituted phenyl ring or C5-C6 monocyclic heteroaryl and a C1-C4 heteroalkylene that is unsubstituted or is substituted with one or two C1-C4 alkyl or heteroalkyl groups, where the alkyl or heteroalkyl groups can optionally cyclize to form a ring such as cyclopropane, dioxolane, or oxacyclopentane.
  • substituents may be on either the alkyl or heteroalkyi portion or on the aryl or heteroaryl portion of the group.
  • the substituents optionally present on the alkyl or heteroalkyi portion are the same as those described above for alkyl groups generally; the substituents optionally present on the aryl or heteroaryl portion are the same as those described above for aryl groups generally.
  • Arylalkyl groups as used herein are hydrocarbyl groups if they are unsubstituted, and are described by the total number of carbon atoms in the ring and alkylene or similar linker.
  • a benzyl group is a C7-arylalkyl group
  • phenylethyl is a C8-arylalkyl.
  • Heteroarylalkyl refers to a moiety comprising an aryl group that is attached through a linking group, and differs from “arylalkyl” in that at least one ring atom of the aryl moiety or one atom in the linking group is a heteroatom selected from N, O and S.
  • the heteroarylalkyl groups are described herein according to the total number of atoms in the ring and linker combined, and they include aryl groups linked through a heteroalkyi linker; heteroaryl groups linked through a hydrocarbyl linker such as an alkylene; and heteroaryl groups linked through a heteroalkyi linker.
  • C7-heteroarylalkyl would include pyridylmethyl, phenoxy, and N- pyrrolylmethoxy.
  • Alkylene refers to a divalent hydrocarbyl group; because it is divalent, it can link two other groups together. Typically it refers to— (CH2)n— where n is 1 -8 and preferably n is 1 -4, though where specified, an alkylene can also be substituted by other groups, and can be of other lengths, and the open valences need not be at opposite ends of a chain.
  • alkylene encompasses more specific examples such as "ethylene,” wherein n is 2, “propylene,” wherein n is 3, and “butylene,” wherein n is 4.
  • the hydrocarbyl groups of the alkylene can be optionally substituted as described above.
  • any alkyl, alkenyl, alkynyl, acyl, or aryl or arylalkyl group that is contained in a substituent may itself optionally be substituted by additional substituents.
  • the nature of these substituents is similar to those recited with regard to the primary substituents themselves if the substituents are not otherwise described.
  • Amino refers to— NH2, but where an amino is described as “substituted” or “optionally substituted”, the term includes NR'R" wherein each R' and R" is independently H, or is an alkyl, alkenyl, alkynyl, acyl, aryl, or arylalkyl group, and each of the alkyl, alkenyl, alkynyl, acyl, aryl, or arylalkyl groups is optionally substituted with the substituents described herein as suitable for the corresponding group; the R' and R" groups and the nitrogen atom to which they are attached can optionally form a 3- to 8-membered ring which may be saturated, unsaturated or aromatic and which contains 1 -3 heteroatoms independently selected from N, 0 and S as ring members, and which is optionally substituted with the substituents described as suitable for alkyl groups or, if NR'R" is an aromatic group, it is optionally
  • carrier refers to a cyclic ring containing only carbon atoms in the ring, whereas the term “carbocycle,” “carbocyclyl,” or “carbocyclic” refers to a cyclic ring containing only carbon atoms in the ring, whereas the term “carbocycle,” “carbocyclyl,” or “carbocyclic” refers to a cyclic ring containing only carbon atoms in the ring, whereas the term “carbocycle,” “carbocyclyl,” or “carbocyclic” refers to a cyclic ring containing only carbon atoms in the ring, whereas the term “carbocycle,” “carbocyclyl,” or “carbocyclic” refers to a cyclic ring containing only carbon atoms in the ring, whereas the term
  • heterocycle or “heterocyclic” refers to a ring comprising a heteroatom.
  • the carbocyclyl can be fully saturated or partially saturated, but non-aromatic.
  • the general term “carbocyclyl” encompasses cycloalkyl.
  • the carbocyclic and heterocyclic structures encompass compounds having monocyclic, bicyclic or multiple ring systems; and such systems may mix aromatic, heterocyclic, and carbocyclic rings. Mixed ring systems are described according to the ring that is attached to the rest of the compound being described.
  • heteroatom refers to any atom that is not carbon or hydrogen, such as nitrogen, oxygen or sulfur, although, in some contexts, “heteroatom” can refer to phosphorus, selenium, or other atoms other than carbon or hydrogen. When it is part of the backbone or skeleton of a chain or ring, a heteroatom must be at least divalent, and will typically be selected from N, O, P, and S.
  • lower alkanoyl refers to an alkanoyl group in which the alkyl portion of the alkanoyl group is C1-C6.
  • the alkyl portion of the alkanoyl group can be optionally substituted as described above.
  • alkylcarbonyl can alternatively be used.
  • alkenylcarbonyl and “alkynylcarbonyl” refer to an alkenyl or alkynyl group, respectively, linked to a carbonyl group.
  • alkoxy refers to an alkyl group covalently linked to an oxygen atom; the alkyl group can be considered as replacing the hydrogen atom of a hydroxyl group.
  • lower alkoxy refers to an alkoxy group in which the alkyl portion of the alkoxy group is C1-C6.
  • the alkyl portion of the alkoxy group can be optionally substituted as described above.
  • haloalkoxy refers to an alkoxy group in which the alkyl portion is substituted with one or more halo groups.
  • sulfo refers to a sulfonic acid (— SO3H) substituent.
  • sulfamoyl refers to a substituent with the structure— S(02)NH2, wherein the nitrogen of the NH2 portion of the group can be optionally substituted as described above.
  • carboxyl refers to a group of the structure— C(0 2 )H.
  • carbamyl refers to a group of the structure— C(02)NH2, wherein the nitrogen of the NH2 portion of the group can be optionally substituted as described above.
  • the terms “monoalkylaminoalkyl” and “dialkylaminoalkyl” refer to groups of the structure— Alki-NH-Alk 2 and — Alki-N(Alk 2 )(Alk3), wherein Alki, Alk2, and Alk3 refer to alkyl groups as described above.
  • alkylsulfonyl refers to a group of the structure — S(0)2-Alk wherein Alk refers to an alkyl group as described above.
  • alkenylsulfonyl and alkynylsulfonyl refer analogously to sulfonyl groups covalently bound to alkenyl and alkynyl groups, respectively.
  • arylsulfonyl refers to a group of the structure— S(0)2-Ar wherein Ar refers to an aryl group as described above.
  • aryloxyalkylsulfonyl refers to a group of the structure— S(0)2-Alk-0-Ar, where Alk is an alkyi group as described above and Ar is an aryl group as described above.
  • arylalkylsulfonyl refers to a group of the structure— S(0)2-AlkAr, where Alk is an alkyi group as described above and Ar is an aryl group as described above.
  • alkyloxycarbonyl refers to an ester substituent including an alkyi group wherein the carbonyl carbon is the point of attachment to the molecule.
  • An example is ethoxycarbonyl, which is ChhChbOCiO)— .
  • alkenyloxycarbonyl refers to similar ester substituents including an alkenyl group, alkenyl group, or cycloalkyl group respectively.
  • aryloxycarbonyl refers to an ester substituent including an aryl group wherein the carbonyl carbon is the point of attachment to the molecule.
  • aryloxyalkylcarbonyl refers to an ester substituent including an alkyi group wherein the alkyi group is itself substituted by an aryloxy group.
  • thiocarbonyl include a carbonyl group in which a double-bonded sulfur replaces the normal double-bonded oxygen in the group.
  • alkylidene and similar terminology refer to an alkyi group, alkenyl group, alkynyl group, or cycloalkyl group, as specified, that has two hydrogen atoms removed from a single carbon atom so that the group is double-bonded to the remainder of the structure.
  • methods and compositions according to the present invention encompass bisantrene derivatives and analogs including one or more optional substituents as defined above, provided that the optionally substituted bisantrene derivative or analog possesses substantially equivalent pharmacological activity to bisantrene as defined in terms of either or both topoisomerase II inhibition and DNA intercalation.
  • Methods for determination of topoisomerase II inhibition are known in the art and are described, for example, in A. Constantinou et al., "Novobiocin- and Phorbol- 12-Myristate-13-Acetate-lnduced Differentiation of Human Leukemia Cells Associates with a Reduction in Topoisomerase II Activity," Cancer Res. 49: 1 1 10-1 1 17 (1989).
  • Methods for determination of DNA intercalation are known in the art and are described, for example, in H. Zipper et al., "Investigations on DNA Intercalation and Surface
  • bisantrene particularly bisantrene dihydrochloride.
  • Liposomes suitable for incorporating the bisantrene or derivative or analog thereof and, in other alternatives as described below, the pyrimidine analog antimetabolite, in a composition according to the present invention can be prepared by techniques well known in the art, including those described in A.S. Janoff, ed.,
  • Liposomes Rational Design (1999, Marcel Dekker, Inc., New York). The use of liposomes to administer antineoplastic agents is disclosed in United States Patent No. 8,750,810 to Okada et al. and in United States Patent No. 9,717,686 to Yang et al.
  • Suitable liposomes include, but are not necessarily limited to, large unilamellar vesicles (LUVs), small unilamellar vesicles (SUVs), and interdigitating fusion liposomes.
  • the liposomes can include a phosphatidylcholine lipid, such as distearylphosphatidylcholine.
  • the liposomes can also include a phosphatidylglycerol lipid, such as
  • the liposomes can also include a sterol such as cholesterol.
  • Liposomes according to the present invention can also include other types of lipids, including, but not limited to, ether lipids, phosphatidic acid, phosphonates, ceramides, ceramide analogs, sphingosines, sphingosine analogs, and serine- containing lipids.
  • Liposomes according to the present invention can also include surface stabilizing hydrophilic polymer-lipid conjugates such as polyethylene glycol- DSPE, to enhance circulation longevity.
  • lipids such as phosphatidylglycerol (PG) and phosphatidylinositol (PI) may also be added to liposome formulations to increase the circulation longevity of the carrier. These lipids may be employed to replace hydrophilic polymer-lipid conjugates as surface stabilizing agents.
  • PG phosphatidylglycerol
  • PI phosphatidylinositol
  • liposomes according to the present invention have a diameter of less than about 300 nm.
  • liposomes according to the present invention have a diameter of less than about 200 nm.
  • encapsulation includes covalent or non-covalent association of an agent with the lipid-based delivery vehicle. For example, this can be by interaction of the agent with the outer layer or layers of the liposome or entrapment of an agent within the liposome, equilibrium being achieved between different portions of the liposome.
  • encapsulation of an agent can be by association of the agent by interaction with the bilayer of the liposomes through covalent or non-covalent interaction with the lipid components or entrapment in the aqueous interior of the liposome, or in equilibrium between the internal aqueous phase and the bilayer.
  • loading or equivalent terminology is used herein to refer to the act of encapsulating one or more agents into a delivery vehicle.
  • Techniques for encapsulation are dependent on the nature of the delivery vehicles.
  • therapeutic agents may be loaded into liposomes using both passive and active loading methods.
  • Passive methods of encapsulating active agents in liposomes involve encapsulating the agent during the preparation of the liposomes. This includes a passive entrapment method described by Bangham et al. (J. Mol. Biol. (1965) 12:238).
  • MLVs multilamellar vesicles
  • LUVs large unilamellar vesicles
  • SUVs small unilamellar vesicles
  • Another suitable method of passive encapsulation includes an ether injection technique described by Deamer and Bangham (Biochim. Biophys. Acta (1976) 443:629) involving dissolving vesicle-forming lipids in ether and, instead of first evaporating the ether to form a thin film on a surface, this film is thereafter put into contact with an aqueous phase to be encapsulated, the ether solution is directly injected into the aqueous phase and the ether is evaporated afterwards, whereby liposomes with encapsulated agents are obtained.
  • Another technique is the Reverse Phase Evaporation technique as described by Szoka and Paphadjopoulos (Proc. Natl. Acad. Sci.
  • lipids in a water insoluble organic solvent is emulsified in an aqueous carrier phase and the organic solvent is subsequently removed under reduced pressure.
  • another suitable method of passive encapsulation involves passive equilibration after the formation of liposomes. This process involves incubating preformed liposomes under altered or non-ambient (based on temperature, pressure, or other factors) conditions and adding a therapeutic agent (e.g., the bisantrene or the derivative or analog thereof and the pyrimidine analog antimetabolite as described in the alternative below) to the exterior of the liposomes. The therapeutic agents then equilibrate into the interior of the liposomes, across the liposomal membrane.
  • a therapeutic agent e.g., the bisantrene or the derivative or analog thereof and the pyrimidine analog antimetabolite as described in the alternative below
  • the liposomes are then returned to ambient conditions and unencapsulated therapeutic agents, if present, are removed via dialysis or another suitable method.
  • Other methods of passive entrapment include subjecting liposomes to successive dehydration and rehydration treatment, or freezing and thawing. Dehydration is carried out by evaporation or freeze-drying. This technique is disclosed by Kirby et al.,
  • One method of pH gradient loading is the citrate-base loading method utilizing citrate as the internal buffer at a pH of 4.0 and a neutral exterior buffer.
  • Other methods employed to establish and maintain a pH gradient across a liposome involve the use of an ionophore that can insert into the liposome membrane and transport ions across membranes in exchange for protons (see U.S. Pat. No. 5,837,282).
  • a recent technique utilizing transition metals to drive the uptake of drugs into liposomes via complexation in the absence of an ionophore may also be used. This technique relies on the formation of a drug-metal complex rather than the establishment of a pH gradient to drive uptake of drug.
  • Other procedures for generation of liposomes are known in the art, including, but not limited to, lipid film/hydration, reverse phase evaporation, detergent dialysis, freeze/thaw, homogenization, solvent dilution, and extrusion.
  • the original external medium can be replaced by a new external medium having a different pH value.
  • the replacement of the external medium can be accomplished by various techniques, such as, by passing the lipid vesicle preparation through a gel filtration column, e.g., a Sephadex G-50 column, which has been equilibrated with the new medium, or by centrifugation, dialysis, or related techniques.
  • the internal medium may be either acidic or basic with respect to the external medium.
  • a pH gradient loadable agent is added to the mixture and encapsulation of the agent in the liposome occurs as described above.
  • a preferred method of pH gradient loading is the citrate-based loading method utilizing citrate as the internal buffer at a pH of 2-6 and a neutral external buffer.
  • Various methods are known in the art for establishing and maintaining a pH gradient across a liposome. This may involve the use of ionophores that can insert into the liposome membrane and transport ions across membranes in exchange for protons. Compounds encapsulated in the interior of the liposome that are able to shuttle protons across the liposomal membrane and thus set up a pH gradient may also be utilized. These compounds comprise an ionizable moiety that is neutral when deprotonated and charged when protonated.
  • the neutral deprotonated form (which is in equilibrium with the protonated form) is able to cross the liposome membrane and thus leave a proton behind in the interior of the liposome and thereby cause a decrease in the pH of the interior by making the interior more acidic.
  • examples of such compounds include methylammonium chloride, methylammonium sulfate, or ethylenediammonium sulfate.
  • Internal loading buffers that are able to establish a basic internal pH can also be utilized. In this case, the neutral form is protonated such that protons are shuttled out of the liposome interior to establish a basic interior.
  • An example of such a compound is calcium acetate.
  • two or more agents may be loaded into a liposome using the same active loading methods or may involve the use of different active loading methods. These alternatives are not required when only the bisantrene or the derivative or analog of bisantrene is to be loaded into the liposome.
  • metal complexation loading may be utilized to actively load multiple agents or may be coupled with another active loading technique, such as pH gradient loading.
  • Metal-based active loading typically uses liposomes with passively encapsulated metal ions (with or without passively loaded therapeutic agents).
  • Various salts of metal ions are used, presuming that the salt is pharmaceutically acceptable and soluble in an aqueous solution.
  • Actively loaded agents are selected based on being capable of forming a complex with a metal ion and thus being retained when so complexed within the liposome, yet capable of loading into a liposome when not complexed to metal ions.
  • Agents that are capable of coordinating with a metal typically comprise coordination sites such as amines, carbonyl groups, ethers, ketones, acyl groups, acetylenes, olefins, thiols, hydroxyl or halide groups or other suitable groups capable of donating electrons to the metal ion thereby forming a complex with the metal ion. Uptake of an agent may be established by incubation of the mixture at a suitable temperature after addition of the agent to the external medium.
  • uptake of the agent may occur over a time period of minutes or hours.
  • Methods of determining whether coordination occurs between an agent and a metal within a liposome include spectrophotometric analysis and other conventional techniques known in the art.
  • liposome loading efficiency and retention properties using metal-based procedures carried out in the absence of an ionophore in the liposome are dependent on the metal employed and the lipid composition of the liposome.
  • loading or retention properties can be tailored to achieve a desired loading or release of a selected agent from a liposome.
  • vesicle-forming lipids which are amphipathic lipids capable of either forming or being incorporated into a bilayer structure.
  • the latter term includes lipids that are capable of forming a bilayer by themselves or when in combination with another lipid or lipids.
  • An amphipathic lipid is incorporated into a lipid bilayer by having its hydrophobic moiety in contact with the interior, hydrophobic region of the membrane bilayer and its polar head moiety oriented toward an outer, polar surface of the membrane. Hydrophilicity may arise from the presence of functional groups such as hydroxyl, phosphato, carboxyl, sulfato, amino or sulfhydryl groups which are polar or substantially polar, including charged groups.
  • lipids included in the liposomes of the invention will typically comprise at least one acyl group with a chain length of at least 16 carbon atoms.
  • phospholipids used as vesicle forming components include dipalmitoyl phosphatidylcholine (DPPC) and distearoyl phosphatidylcholine (DSPC).
  • DPPC dipalmitoyl phosphatidylcholine
  • DSPC distearoyl phosphatidylcholine
  • DPPC dipalmitoyl phosphatidylcholine
  • DSPC distearoyl phosphatidylcholine
  • DPPC is a common saturated chain (Cie) phospholipid with a bilayer phase transition temperature of 41 .5° C.
  • DPPC 1 ,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1 -glycerol)]
  • DSPC 1 ,2-distearoyl-sn-glycero-3-phosphocholine
  • the liposomes of the present invention have a phase transition temperature greater than 38° C; this can be accomplished by employing components which confer this property.
  • the ultimate transition temperature will depend on the acyl chain length as well as the degree of unsaturation of the acyl groups. Including unsaturation in the chain lowers the transition temperature so that in the event the acyl groups are unsaturated, acyl groups containing 18 carbons or 20 carbons or more can be used. Liposomes may also be prepared such that the liquid crystalline transition temperature is greater than 45° C.
  • Vesicle-forming lipids making up the liposome are phospholipids such as phosphatidylcholine (PC), phosphatidyl (PA) or phosphatidylethanolamine (PE), containing two saturated fatty acids, while the acyl chains are preferably stearoyl (18:0), nonadecanoyl (19:0), arachidoyl (20:0),
  • PC phosphatidylcholine
  • PA phosphatidyl
  • PE phosphatidylethanolamine
  • heniecosanoyl (21 :0), behenoyl (22:0), tricosanoyl (23:0), lignoceroyl (24:0) or cerotoyl (26:0); other lipids can also be incorporated.
  • a heterodisperse suspension of liposomes formed by methods described above may be "size reduced” using conventional techniques to produce liposomes within a desired size range and reduced polydispersity.
  • Conventional size-reduction techniques include but are not limited to sonication, homogenization and extrusion.
  • extrusion methods a variety of membrane pore sizes are available to produce liposomes in various size ranges.
  • a drawback of this technique for low-cholesterol liposomes is the tendency for large vesicles to deform and thus pass through narrow extrusion filters when extruded under standard temperatures (i.e., above the vesicle phase transition temperature, which is a temperature above the phase transition temperature of the highest melting lipid in the lipid-based delivery vehicle).
  • a heterodisperse suspension of MLVs is extruded at least once at the higher
  • the liposomes are low-cholesterol liposomes.
  • the incorporation of less than 20 mol % cholesterol in liposomes can allow for retention of drugs not optimally retained when liposomes are prepared with greater than 20 mol % cholesterol. Additionally, liposomes prepared with less than 20 mol % cholesterol display narrow phase transition
  • Liposomes may also be prepared with surface stabilizing hydrophilic polymer-lipid conjugates such as polyethylene glycol-DSPE, to enhance circulation longevity.
  • hydrophilic polymer-lipid conjugates such as polyethylene glycol-DSPE
  • PG phosphatidylglycerol
  • PI phosphatidylinositol
  • Cholesterol-free liposomes containing PG or PI to prevent aggregation may be prepared, thereby increasing the blood residence time of the carrier.
  • Suitable proportions of lipids used in liposomes according to the present invention are as described below.
  • the ratio of the pyrimidine analog antimetabolite and the bisantrene or derivative or analog thereof can be from about 25: 1 to about 1 : 1 .
  • the ratio of the pyrimidine analog antimetabolite and the bisantrene or derivative or analog thereof is from about 10: 1 to about 3: 1. More preferably, the ratio of the pyrimidine analog antimetabolite and the bisantrene or derivative or analog thereof is about 5: 1 .
  • the liposomes can include a negatively charged lipid having a hydrophilic portion and a hydrophobic portion with a neutral non-zwitterionic moiety attached to the hydrophilic portion of the lipid.
  • the negatively charged lipid is typically a phospholipid or a sphingophospholipid.
  • the lipid is a
  • the non-zwitterionic moiety is attached to this negatively charged lipid, preferably to the phosphate group.
  • the non- zwitterionic moiety is neutral such that the net negative charge on a lipid used in this invention is due solely to the negative charge of the lipid component.
  • the non- zwitterionic moiety may comprise functional groups that impart a desired hydrophilicity to the lipid, such groups being selected from alcohols, ketones, carboxylic acids, ethers and amines.
  • a preferred non-zwitterionic moiety is a short-chain alcohol such as glycerol, or a cyclic alcohol, such as inositol, or a polyalkylene oxide such as PEG.
  • the liposomes comprise at least 10% of such negatively charged lipids having a hydrophilic portion and a hydrophobic portion with a neutral non- zwitterionic moiety attached to the hydrophilic portion of the lipid.
  • the liposomes can have an intraliposomal osmolality of 500 mOSM/kg or less.
  • the liposome comprises distearoylphosphatidylcholine, distearoylphosphatidylglycerol and cholesterol.
  • the molar ratio of the distearoylphosphatidylcholine, the distearoylphosphatidylglycerol, and the cholesterol is from about 6.5 to about 7.5 of distearoylphosphatidylcholine, about 1 .5 to about 2.5 of distearoylphosphatidylglycerol, and about 0.8 to 1 .2 of cholesterol.
  • the molar ratio of the distearoylphosphatidylcholine, the distearoylphosphatidylglycerol, and the cholesterol is from about 6.8 to about 7.2 of distearoylphosphatidylcholine, about 1 .8 to about 2.2 of distearoylphosphatidylglycerol, and about 0.9 to 1 .1 of cholesterol. More preferably, in this alternative, the molar ratio of the distearoylphosphatidylcholine, the distearoylphosphatidylglycerol, and the cholesterol is about 7:2: 1 .
  • Liposomes suitable for use as part of compositions according to the present invention are disclosed in the following United States Patents: 7,850,990 to Tardi et al.; 8,022,279 to Mayer et al.; 8,431 ,806 to Mayer et al.; 8,518,437 to Tardi et al.; and 9,271 ,931 to Tardi et al., and also in the following United States Patents: 7,850,990 to Tardi et al.; 8,022,279 to Mayer et al.; 8,431 ,806 to Mayer et al.; 8,518,437 to Tardi et al.; and 9,271 ,931 to Tardi et al., and also in the following United States Patents: 7,850,990 to Tardi et al.; 8,022,279 to Mayer et al.; 8,431 ,806 to Mayer et al.; 8,518,437 to Tardi
  • composition of matter comprising: (i) a therapeutically effective quantity of bisantrene or a derivative or analog thereof; (ii) a therapeutically effective quantity of a pyrimidine analog antimetabolite; and (iii) a liposome encapsulating both the therapeutically effective quantity of bisantrene or the derivative or analog thereof and the pyrimidine analog antimetabolite.
  • the bisantrene or derivative or analog thereof is as described above.
  • the liposome is also as described above, with the limitation that the liposome encapsulates both the therapeutically effective quantity of bisantrene or the derivative or analog thereof and the pyrimidine analog antimetabolite.
  • Pyrimidine analog antimetabolites that can be included in this alternative according to compositions according to the present invention include cytarabine, 5- azacytidine, gemcitabine, decitabine, fluoropyrimidines, 2'-deoxy-2'- methylidenecytidines, and analogs and derivatives as described below, as well as other pyrimidine analog antimetabolites as described below.
  • the pyrimidine analog antimetabolite is typically selected from the group consisting of cytarabine, 5- azacytidine, gemcitabine, floxuridine, 5-fluorouracil, capecitabine, 6-azauracil, troxacitabine, thiarabine, sapacitabine, CNDAC (2'-cyano-2'-deoxy-1 - ⁇ -D- arabinofuranosylcytosine), 2'-deoxy-2'-methylidenecytidine, 2'-deoxy-2'- fluoromethylidenecytidine, 2'-deoxy-2'-methylidene-5-fluorocytidine, 2'-deoxy-2',2'- difluorocytidine, and 2'-C-cyano-2'-deoxy- -arabinofuranosylcytosine.
  • the pyrimidine analog antimetabolite is selected from the group consisting of cytarabine, 5- azacytidine, gemcitabine, floxuridine, 5-fluorouracil, capecitabine, and 6-azauracil. More preferably, the pyrimidine analog antimetabolite is cytarabine.
  • Cytarabine has the structure shown as Formula (N-l):
  • Cytarabine (4-amino-1 -[(2R,3S,4R,5R)-3,4-dihydroxy-5- (hydroxymethyl)oxolan-2-yl] pyrimidin-2-one) is a pyrimidine nucleoside antimetabolite.
  • the compound is an analog of 2'-deoxycytidine with the 2'-hydroxyl in a position trans to the 3'-hydroxyl of the sugar moiety; therefore, the sugar moiety is an arabinose moiety rather than a ribose moiety.
  • Cytarabine must be activated via conversion of the 5'-monophosphate nucleotide (AraCMP) to terminate strand synthesis.
  • AraCMP is then able to react with nucleotide kinases to form diphosphate and triphosphate nucleotides (AraCDP and AraCTP).
  • the incorporation of cytarabine into DNA is S-phase specific. Cytarabine inhibits DNA synthesis and kills cells, particularly rapidly dividing cells. It also inhibits DNA and RNA polymerases, as well as nucleotide reductases. Resistance to cytarabine can develop. Cytarabine can be rapidly deaminated by the enzyme cytidine deaminase in serum into the inactive uracil derivative.
  • AraCMP is also deaminated by deoxycytidylate deaminase, leading to the inactive uridine-5'- monophosphate analog. Additionally, AraCTP is also a substrate for SAMDH1 , which hydrolyzes 2'-deoxynucleoside-5'-triphosphates (dNTPs) into 2'-deoxynucleosides and inorganic triphosphate products. Cytarabine can exhibit toxicity, particularly
  • cytarabine can produce leukopenia, thrombocytopenia, and anemia with megaloblastic changes. Gastrointestinal disturbances, fever, conjunctivitis, pneumonitis, hepatic dysfunction, dermatitis, and neurotoxicity have also been noted, particularly at higher dosages.
  • nucleoside analog 5-azacytidine has the structure shown as Formula
  • the analog 5-azacytidine (4-amino-1 -p-D-ribofuranosyl-1 ,3,5-triazin- 2(1 /-/)-one) inhibits DNA methyltransferase, causing hypomethylation of DNA, and also is incorporated directly into both RNA and DNA resulting in cell death; 5-azacytidine is incorporated into RNA more frequently than into DNA.
  • the analog 5-azacytidine is used in the treatment of myelodysplastic syndrome and acute myeloid leukemia. It has a number of side effects, including anemia, neutropenia, thrombocytopenia,
  • nucleoside analog gemcitabine has the structure shown as Formula
  • Gemcitabine is 4-amino-1 -(2-deoxy-2,2-difluoro- -D-eryi/?ro- pentofuranosyl)pyrimidin-2(1 /-/)-one.
  • Gemcitabine is used for treating pancreatic and in combination with cisplatin for advanced or metastatic bladder cancer and advanced or metastatic non-small cell lung cancer, as well as in various combinations for ovarian cancer or breast cancer.
  • Gemcitabine is hydrophilic and is transported into cells via molecular transporters for nucleosides, particularly SLC29A1 SLC28A1 , and SLC28A3. Gemcitabine is then phosphorylated to gemcitabine monophosphate
  • gemcitabine then is incorporated into DNA in place of cytidine.
  • gemcitabine When gemcitabine is incorporated into DNA it allows a native, or normal, nucleoside base to be added next to it. This leads to "masked chain termination” as gemcitabine is a "faulty" base, but due to its neighboring native nucleoside it eludes the cell's normal repair system (base-excision repair).
  • base-excision repair base-excision repair
  • incorporation of gemcitabine into the cell's DNA creates an irreparable error that leads to inhibition of further DNA synthesis, and thereby leading to cell death.
  • Gemcitabine like many other inhibitors of DNA replication, can inhibit bone marrow function and cause anemia, neutropenia, and thrombocytopenia. It also may have other side effects, particularly affecting the respiratory system.
  • Floxuridine is another nucleoside analog.
  • Floxuridine has the structure shown as Formula ( -IV):
  • Floxuridine is 5-fluoro-1 -[4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2- yl]-1 /-/-pyrimidine-2,4-dione.
  • Floxuridine is used for the treatment of colorectal cancer, kidney cancer, and stomach cancer.
  • Floxuridine is converted in vivo to 5-fluorouracil, which interferes with both DNA and RNA synthesis; 5-fluorouracil also inhibits uracil riboside phosphorylase, which prevents the utilization of preformed uracil in RNA synthesis.
  • the side effects of floxuridine are similar to the side effects of other pyrimidine analog antimetabolites.
  • the compound 5-fluorouracil is used to treat esophageal cancer, colon cancer, stomach cancer, pancreatic cancer, breast cancer, and cervical cancer, among others. It has the lUPAC name of 5-fluoro-1 H,3H-pyrimidine-2,4-dione and blocks the action of thymidylate synthase, inhibiting DNA synthesis.
  • the side effects of 5- fluorouracil are generally similar to the side effects of other pyrimidine antimetabolites.
  • Capecitabine is another pyrimidine antimetabolite. Capecitabine has the structure shown as Formula (N-VI):
  • Capecitabine is used to treat breast cancer, gastric cancer, and
  • pyrimidine antimetabolites include 6-azauracil and the additional pyridine antimetabolites disclosed in W.B. Parker, "Enzymology of Purine and
  • the compound 6-azauracil has the structure shown in Formula (N-VII):
  • Troxacitabine has the structure shown in Formula (N-VII I):
  • Thiarabine has the structure shown in Formula (N-IX):
  • Sapacitabine has the structure shown in Formula (N-X):
  • N-X has the lUPAC name 1 -(2-cyano-2-deoxy-p-D-arabinofuranosyl)-4- (palmitoylamino)pyrimidin-2(1 /-/)-one. It is a prodrug of CNDAC (2'-cyano-2'-deoxy-1 - ⁇ - D-arabinofuranosylcytosine).
  • CNDAC (2'-cyano-2'-deoxy-1 - -D-arabinofuranosylcytosine) has the structure shown in Formula -XI):
  • Still other pyrimidine analog antimetabolites are known in the art, including 2'-deoxy-2'-methylidenecytidine compounds as disclosed in United States Patent No. 5,776,488 to Mori et al., including 2'-deoxy-2'-methylidenecytidine, 2'-deoxy- 2'-fluoromethylidenecytidine, 2'-deoxy-2'-methylidene-5-fluorocytidine, 2'-deoxy-2',2'- difluorocytidine, and 2'-C-cyano-2'-deoxy- -arabinofuranosylcytosine. Additional pyrimidine analog antimetabolites are also known in the art. [0147] Liposomes suitable for incorporating the bisantrene or derivative or analog thereof and the pyrimidine analog antimetabolite in this aspect of the invention can be prepared by techniques well known in the art and described above.
  • compositions according to the present invention as described above can be administered to treat malignancies, including, but not limited to, breast cancer, ovarian cancer, renal cancer, small-cell lung cancer, non-small cell lung cancer,
  • a method of treating a malignancy comprises administration of a
  • composition comprises a therapeutically effective quantity of bisantrene or a derivative or analog thereof as described above.
  • composition comprises a therapeutically effective quantity of bisantrene or a derivative or analog thereof and a pyrimidine analog antimetabolite as described above.
  • the breast cancer can be selected from the group consisting of refractory breast cancer, triple-negative breast cancer, and breast cancer characterized by overexpressed Her-2-neu.
  • the cancer is an acute leukemia of childhood
  • the acute leukemia of childhood can be selected from the group consisting of acute myelocytic leukemia (AML) and acute lymphocytic leukemia (ALL) of childhood.
  • the cancer is prostate cancer
  • the prostate cancer can be androgen- resistant prostate cancer.
  • the cancer is glioblastoma
  • the glioblastoma can be glioblastoma that is resistant to one or both of the following therapeutic agents:
  • temozolomide Temodar
  • bevacizumab Avastin
  • EGFR Variant III EGFR Variant III
  • the cancer can be a malignancy characterized by
  • compositions according to the present invention can also be used to treat other hyperproliferative diseases, including myelodysplastic syndrome and mycosis fungoides.
  • compositions according to the present invention can be administered to humans or to socially or economically important non-human species such as dogs, cats, pigs, sheep, goats, rabbits, cattle, or horses. Unless specifically stated, methods as described below are not limited to treatment of humans.
  • compositions according to the present invention should be utilized with respect to dose, schedule and route of administration using established protocols.
  • applications may also utilize dose escalation should agents encapsulated in delivery vehicle compositions of the present invention exhibit reduced toxicity to healthy tissues of the subject.
  • compositions according to the present invention are:
  • parenterally i.e., intraarterially, intravenously, intraperitoneally
  • the pharmaceutical compositions are administered intravenously or intraperitoneally by a bolus or infusional injection.
  • a bolus or infusional injection For example, see Rahman et al., U.S. Pat. No. 3,993,754; Sears, U.S. Pat. No. 4, 145,410; Papahadjopoulos et al., U.S. Pat. No. 4,235,871 ; Schneider, U.S. Pat. No. 4,224, 179; Lenk et al., U.S. Pat. No. 4,522,803; and Fountain et al., U.S. Pat. No. 4,588,578.
  • compositions according to the present invention can be contacted with the target tissue by direct application of the preparation to the tissue.
  • the application may be made by topical, "open,” or “closed” procedures.
  • topical means the direct application of the multi-drug preparation to a tissue exposed to the environment, such as the skin, oropharynx, external auditory canal, and the like.
  • Open procedures are those procedures that include incising the skin of a patient and directly visualizing the underlying tissue to which the
  • compositions according to the present invention are applied. This is generally accomplished by a surgical procedure, such as a thoracotomy to access the lungs, abdominal laparotomy to access abdominal viscera, or other direct surgical approach to the target tissue. "Closed" procedures are invasive procedures in which the internal target tissues are not directly visualized, but accessed via inserting instruments through small wounds in the skin.
  • compositions according to the present invention may be administered to the peritoneum by needle lavage.
  • compositions according to the present invention may be administered through endoscopic devices.
  • compositions according to the present invention can be prepared according to standard techniques and may comprise water, buffered water, 0.9% saline, 0.3% glycine, 5% dextrose, iso-osmotic sucrose solutions and the like, including glycoproteins for enhanced stability, such as albumin, lipoprotein, globulin, and the like. These compositions may be sterilized by conventional, well-known sterilization techniques. The resulting aqueous solutions may be packaged for use or filtered under aseptic conditions and lyophilized, the lyophilized preparation being combined with a sterile aqueous solution prior to administration.
  • the compositions may contain pharmaceutically acceptable auxiliary substances as required to approximate
  • physiological conditions such as pH adjusting and buffering agents, tonicity adjusting agents and the like, for example, sodium acetate, sodium lactate, sodium chloride, potassium chloride, calcium chloride, and the like.
  • suitable carriers can include water, buffered water, 5% dextrose, 0.4% saline or 0.9% saline, 0.3% glycine, and proteins for enhanced stability, including albumin, lipoproteins, glycoproteins, or globulin in order to produce a pharmaceutical composition suitable for administration by the selected route of administration as described above.
  • the delivery vehicle suspension may include lipid-protective agents which protect lipids against free- radical and lipid-peroxidative damages on storage.
  • compositions according to the present invention may further contain pharmaceutically acceptable auxiliary substances as required to approximate physiological conditions, such as pH adjusting and buffering agents, tonicity adjusting agents and the like, for example, sodium acetate, sodium lactate, sodium chloride, potassium chloride, calcium chloride, and the like.
  • Leucovorin may also be administered with compositions of the invention through standard techniques to enhance the life span of administered fluoropyrimidines when a composition according to the present invention includes a fluoropyrimidine.
  • the concentration of delivery vehicles in the pharmaceutical formulations can vary widely, such as from less than about 0.05%, usually at or at least about 2-5% to as much as 10 to 30% by weight and will be selected primarily by fluid volumes, viscosities, and the like, in accordance with the particular mode of administration selected. For example, the concentration may be increased to lower the fluid load associated with treatment. Alternatively, delivery vehicles composed of irritating lipids may be diluted to low concentrations to lessen inflammation at the site of administration. For diagnosis, the amount of delivery vehicles administered will depend upon the particular label used, the disease state being diagnosed and the judgment of the clinician.
  • compositions according to the present invention are administered intravenously.
  • routes of administration particularly parenteral administration, such as intraperitoneal administration, can alternatively be used.
  • Dosage for the delivery vehicle formulations will depend on the ratio of drug to lipid and the administrating physician's opinion based on age, weight, and condition of the patient as well as other drugs being administered and
  • kidney and liver function pharmacokinetic considerations such as kidney and liver function.
  • preventative measures wherein the object is to prevent or slow down (lessen) an undesired physiological change or disorder, such as the growth, development, or spread of cancer.
  • beneficial or desired clinical results include, but are not limited to, alleviation of symptoms, diminishment of extent of disease, stabilized (i.e., not worsening) state of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, and remission (whether partial or total), whether detectable or undetectable.
  • Treatment can also mean prolonging survival as compared to expected survival if not receiving treatment.
  • Those in need of treatment include those already with the condition or disorder as well as those prone to have the condition or disorder or those in which the condition or disorder is to be prevented.
  • the results of treatment can be determined by methods known in the art, such as determination of reduction of pain as measured by reduction of requirement for administration of opiates or other pain medication, determination of reduction of tumor burden, determination of restoration of function as determined by an improvement in the Karnofsky Performance Score, or other methods known in the art.
  • the use of terms such as “treat” or “treatment” is not to be understood as implying a cure for any disease or condition.
  • compositions according to the present invention may be measured after administration into an animal model.
  • the animal model comprises a tumor although delivery vehicle compositions may be administered to animal models of other diseases.
  • Rodent species such as mice and rats of either inbred, outbred, or hybrid origin including immunocompetent and immunocompromised, as well as knockout, or transgenic models may be used.
  • Models can consist of solid or non-solid tumors implanted as cell suspensions, bries or tumor fragments in either subcutaneous, intravenous, intraperitoneal, intramuscular, intrathecal, or orthotopic regions. Tumors may also be established via the application or administration of tumorigenic/carcinogenic agents or may be allowed to arise spontaneously in
  • Tumor types can consist of tumors of ectodermal, mesodermal, or endodermal origin such as carcinomas, sarcomas, melanomas, gliomas, leukemias and lymphomas.
  • mouse models of tumors are employed.
  • Human xenograft solid tumors grown in immune compromised mice may be utilized and selected on the basis of defined genetics and growth attributes.
  • Tumor cells utilized in these experiments can be genetically manipulated or selected to express preferable properties and are injected into mice. Once the tumors have grown to a palpable (measurable) size, delivery vehicle compositions can be administered, preferably intravenously, and their effects on tumor growth are monitored. Intended therapeutic treatments can consist of single bolus or push administrations or multiple or continuous administrations over several days or weeks and by any
  • syringes such as by the oral, nasal, subcutaneous, intravenous, intraperitoneal, intrathecal, intratumoral, or other appropriate routes using syringes, tablets, liquids, and pumps (such as osmotic). Dose and schedule dependency may be evaluated in order to determine the maximum anti-tumor activity that can be achieved.
  • Solid tumor model evaluation methods include measurement of tumor volume (mass), tumor weight inhibition (TWI %), tumor growth delay (T-C), tumor regression, cell kill and clonogenic assays. Tumor volume measurements are determined from vernier caliper measurements of
  • Tumor weight inhibition (TWI %) is determined by measuring the mean tumor weight of a treated group divided by the mean tumor weight of a control group, minus 1 x 100 at a defined time point.
  • Tumor growth delay (T-C) is measured as the median time in days for a treated group (T) to reach an arbitrarily determined tumor size (for example, 300 mg) minus median time in days for the control group to reach the same tumor size.
  • Tumor regression as a result of treatment may also be used as a means of evaluating a tumor model. Results are expressed as reductions in tumor size (mass) over time.
  • Cell kill methods of solid tumor model evaluation can involve measuring tumors repeatedly by calipers until all exceed a predetermined size (e.g., 200 mg). The tumor growth and tumor doubling time can then be evaluated.
  • Clonogenicity assays express the effectiveness of therapy. These assays include excision assays and characterization of cell suspensions from solid tumors. Excision assays, used to assess what fraction of cells, in a suspension prepared from tumors, have unlimited proliferative potential (i.e., are clonogenic).
  • excision assays Three types of excision assays are: (i) TD50, or endpoint dilution assays, which determine the number of cells required to produce tumor takes from inocula in vivo; (ii) in vivo colony assays, which assess the ability of individual cells to form nodules (colonies) in, for example, the lung; and (iii) in vitro colony assay, which test the ability of individual cells to grow into colonies either in liquid media, when colonies form on the plastic or glass surface of culture dishes, or in semisolid media such as agar, in which the colonies form in suspension.
  • Characterization of cell suspensions from solid tumors are required for in vitro and in vivo clonogenic assays, flow-cytometric measurements, and for numerous biochemical and molecular analyses performed on a per cell basis. Preparation is by a number of methods such as enzymatic, mechanical, chemical, combinations thereof, and surface activity agents. Evaluations could include, cell yield, cell morphology, tumor cell clonogenicity, retention of biochemical or molecular characteristics.
  • Non- solid tumor model evaluation methods include measurement of increase in life span (ILS %), tumor growth delay (T-C), long-term survivors (cures). Increase in life-span (ILS %) measures the percentage increase in life-span of treated groups versus control or untreated groups.
  • Tumor growth delay measures median time in days for treated (T) group survival minus median time in days for control (C) group survival.
  • Long-term survivors measures treatment groups that survive up to and beyond 3-times the survival times of untreated or control groups.
  • Methods of determining therapeutic activity in humans afflicted with cancer include measurements of survival and surrogate endpoints. The time at which survival is reasonably evaluated depends on the tumor in question. By way of example, survival rates for patients with low-grade lymphomas may be examined at 5 or 10 years post diagnosis, whereas the survival or patients having aggressive diseases such as advanced non-small cell lung cancer may be best evaluated at 6 or 12 months post diagnosis.
  • CR complete response
  • PR partial response
  • PFS progression-free survival
  • TTP time-to-progression
  • DOR duration of response
  • plasma and urine markers enzyme inhibition and/or receptor status, changes in gene expression and quality of life (QOL).
  • a complete response means the disappearance of all known sites of disease without the development of any new disease for a period of time appropriate for the tumor type being treated.
  • Assessments are based on a variety of examinations such as those stated above. Partial response is at least a 50% decrease in the sum of the products of the bidimensional measurement of all lesions with no new disease appearing for a period of time appropriate for the tumor type being treated.
  • Progression-free Survival Duration from treatment in which a patient survives and there is no growth of existing tumor nor appearance of new tumor masses. PFS may be expressed as either the duration of time or as the proportion of patients who are surviving and progression-free at a given time after diagnosis.
  • Time-to-progression TTP
  • duration of response DOR
  • Plasma and urine markers include measuring markers such as, but not limited to, the following markers: prostate specific antigen (PSA) and carcinoembryonic antigen (CEA).
  • PSA prostate specific antigen
  • CEA carcinoembryonic antigen
  • Enzyme inhibition and/or receptor status can include assessment of growth factor receptors such as, but not limited to, tyrosine kinase receptors, EGF receptor, PDGF receptor, Her-1 and Her-2 receptors, or assessment of enzymes such as, but not limited to, integrin-linked kinases, protein kinases and the like. Changes in gene expression include serial analysis of gene expression (genomics) and changes in protein
  • QOL Quality of Life
  • the measure also yields single-item ratings of additional symptoms commonly reported by cancer patients (dyspnea, appetite loss, sleep disturbance, constipation, and diarrhea) as well as the perceived financial impact of the disease and its treatment.
  • Methods according to the present invention can further comprise administration of a therapeutically effective quantity of an additional therapeutic agent to treat the disease, disorder, or condition, in particular, a malignancy as described above.
  • the additional therapeutic agent can be selected from the group consisting of tamoxifen, anastrozole, letrozole, cyclophosphamide, docetaxel, paditaxel, methotrexate, fluorouracil, and trastuzumab.
  • the additional agent can be selected from the group consisting of: platinum-containing antineoplastic drugs such as cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatin, phenanthriplatin, picoplatin, and satraplatin; paditaxel; topotecan; gemcitabine; etoposide; and
  • the additional agent can be selected from the group consisting of everolimus, torisel, nexavar, sunitinib, axitinib, inferferon, interleukin-2, pazopanib, sorafenib, nivolumab, cabozanitib, and levanitib.
  • the additional agent can be selected from the group consisting of cyclophosphamide, cisplatin, etoposide, vincristine, paditaxel, and carboplatin.
  • the additional agent can be selected from the group consisting of cisplatin, erlotinib, gefitinib, afatinib, crizotinib, bevacizumab, carboplatin, paditaxel, nivolumab, and pembrolizumab.
  • the additional agent can be selected from the group consisting of mechlorethamine, vincristine, prednisone, procarbazine, bleomycin, vinblastine, dacarbazine, etoposide, and cyclophosphamide.
  • the additional agent can be selected from the group consisting of cyclophosphamide, vincristine, and prednisone.
  • the additional agent can be selected from the group consisting of cytarabine (provided that the liposomal
  • composition does not include cytarabine), fludarabine, all-frans-retinoic acid, interleukin- 2, and arsenic trioxide.
  • the additional agent can be selected from the group consisting of temozolomide,
  • the additional agent can be selected from the group consisting of 5- fluorouracil (provided that the liposomal composition does not include 5-fluorouracil), capecitabine (provided that the liposomal composition does not include capecitabine), carmustine, semustine, mitomycin C, cisplatin, taxotere, and trastuzumab.
  • the additional agent can be selected from the group consisting of mitotane, cisplatin, etoposide, and streptozotocin.
  • the additional agent can be selected from the group consisting of paclitaxel, carboplatin, cetuximab, docetaxel, cisplatin, and 5-fluorouracil (provided that the liposomal composition does not include 5- fluorouracil).
  • the additional agent can be selected from the group consisting of tamoxifen, octreoside, synthetic retinoids, cisplatin, 5-fluorouracil (provided that the liposomal composition does not include 5-fluorouracil), interferon, taxol, and sorafenib.
  • the additional agent can be selected from the group consisting of nivolumab, everolimus, sorafenib, axitinib, lenvatinib, temsirolimus, sunitinib, pazopanib, interleukin-2, cabozanitib, bevacizumab, interferon a, ipilimumab, atezolizumab, varilumab, durvalumab, tremelimumab, and avelumab.
  • the additional agent can be selected from the group consisting of cisplatin, 5-fluorouracil (provided that the liposomal composition does not include 5-fluorouracil), mitomycin C, gemcitabine (provided that the liposomal composition does not include gemcitabine), methotrexate, vinblastine, carboplatin, paclitaxel, docetaxel, ifosfamide, and pemetrexed.
  • the additional agent can be selected from the group consisting of methotrexate, nelarabine, asparaginase, blinatumomab, cyclophosphamide, clofarabine, cytarabine (provided that the liposomal composition does not include cytarabine), dasatinib, methotrexate, imatinib, pomatinib, vincristine, 6-mercaptopurine, pegaspargase, and prednisone.
  • the additional agent can be selected from the group consisting of asparaginase, vincristine, dexamethasone, methotrexate, 6-mercaptopurine, cytarabine (provided that the liposomal composition does not include cytarabine), hydrocortisone, 6-thioguanine, prednisone, etoposide, cyclophosphamide, mitoxantrone, and teniposide.
  • the additional agent can be selected from the group consisting of fludarabine, cyclophosphamide, rituximab, vincristine, prednisolone, bendamustine, alemtuzumab, ofatumumab, obinutuzumab, ibrutinib, idelalisib, and venetoclax.
  • the additional agent can be selected from the group consisting of temozolomide, docetaxel, cabazitaxel, bevacizumab, thalidomide, prednisone, sipuleucel-T, abiraterone, and enzalutamide.
  • the additional agent can be selected from the group consisting of temozolomide and bevacizumab.
  • the additional agent can selected from the group consisting of bortezomib, lenalidomide, dexamethasone, melphalan, prednisone, thalidomide, and cyclophosphamide.
  • Methods for administration of the additional agents are known in the art. Typically, they are administered in a separate pharmaceutical composition according to methods well known in the art, with suitable carriers, excipients, and other components as generally used in the art, although, in some cases, in which the additional agent or agents are compatible with a liposome composition according to the present invention as described above, they may be administered in the liposome composition.
  • pyrimidine analog antimetabolites such as cytarabine
  • compositions according to the present invention possess industrial applicability as pharmaceutical compositions.
  • the methods of the present invention provide specific method steps that are more than general applications of laws of nature and require that those practicing the method steps employ steps other than those conventionally known in the art, in addition to the specific applications of laws of nature recited or implied in the claims, and thus confine the scope of the claims to the specific applications recited therein. In some contexts, these claims are directed to new ways of using an existing drug.

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Abstract

La présente invention concerne des compositions comprenant, dans un liposome, du bisantrène ou un dérivé ou analogue de celui-ci, ainsi que des compositions comprenant, dans un liposome, du bisantrène ou un dérivé ou analogue de celui-ci et un antimétabolite analogue de la pyrimidine tel que la cytarabine, ainsi que des procédés d'utilisation de ces compositions, en particulier dans le traitement de malignités. Les compositions améliorent la stabilité et la biodisponibilité du bisantrène ou du dérivé ou analogue de celui-ci et réduisent la probabilité d'apparition et la gravité d'effets secondaires, tels qu'une phlébite et d'autres complications touchant le système circulatoire, de l'administration du bisantrène ou du dérivé ou analogue de celui-ci et fournissent également une manière efficace d'administrer au moins deux agents antinéoplasiques, lorsque l'antimétabolite analogue de la pyrimidine est inclus dans la composition, dans des quantités thérapeutiquement efficaces pour le traitement d'un certain nombre de types de malignités.
PCT/IB2018/001266 2017-10-13 2018-10-09 Formulations liposomales de bisantrène ou de dérivés ou analogues de celui-ci WO2019073296A1 (fr)

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2021016464A1 (fr) * 2019-07-23 2021-01-28 City Of Hope Méthodes et compositions pour le traitement du cancer
WO2022159497A1 (fr) * 2021-01-20 2022-07-28 Board Of Regents, The University Of Texas System Programmes de polythérapie pour traiter le cancer
WO2023245248A1 (fr) * 2022-06-22 2023-12-28 Race Oncology Ltd Traitement du mélanome
EP4183397A4 (fr) * 2020-07-17 2024-03-27 Delta-Fly Pharma, Inc. Nouvelle méthode thérapeutique et nouvel agent thérapeutique pour le cancer hématologique

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DE3825374A1 (de) * 1988-07-26 1990-02-01 Schwendener Reto Dipl Apotheke Komplex aus mindestens einer lipophilen saeure und mitoxantron und/oder bisantren
EP2520281A2 (fr) * 2009-12-29 2012-11-07 Kyungpook National University Industry- Academic Cooperation Foundation Système d'administration ciblée de médicaments pour le diagnostic et le traitement du cancer contenant un liposome marqué par des peptides qui cible spécifiquement les récepteurs d'interleukine-4, et son procédé de fabrication
WO2015013579A1 (fr) * 2013-07-26 2015-01-29 Update Pharma Inc. Compositions permettant d'améliorer l'avantage thérapeutique du bisantrène

Patent Citations (3)

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Publication number Priority date Publication date Assignee Title
DE3825374A1 (de) * 1988-07-26 1990-02-01 Schwendener Reto Dipl Apotheke Komplex aus mindestens einer lipophilen saeure und mitoxantron und/oder bisantren
EP2520281A2 (fr) * 2009-12-29 2012-11-07 Kyungpook National University Industry- Academic Cooperation Foundation Système d'administration ciblée de médicaments pour le diagnostic et le traitement du cancer contenant un liposome marqué par des peptides qui cible spécifiquement les récepteurs d'interleukine-4, et son procédé de fabrication
WO2015013579A1 (fr) * 2013-07-26 2015-01-29 Update Pharma Inc. Compositions permettant d'améliorer l'avantage thérapeutique du bisantrène

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2021016464A1 (fr) * 2019-07-23 2021-01-28 City Of Hope Méthodes et compositions pour le traitement du cancer
EP4183397A4 (fr) * 2020-07-17 2024-03-27 Delta-Fly Pharma, Inc. Nouvelle méthode thérapeutique et nouvel agent thérapeutique pour le cancer hématologique
US11957701B2 (en) 2020-07-17 2024-04-16 Delta-Fly Pharma, Inc. Therapy and new therapeutic agent for blood cancer
WO2022159497A1 (fr) * 2021-01-20 2022-07-28 Board Of Regents, The University Of Texas System Programmes de polythérapie pour traiter le cancer
WO2023245248A1 (fr) * 2022-06-22 2023-12-28 Race Oncology Ltd Traitement du mélanome
WO2023245249A1 (fr) * 2022-06-22 2023-12-28 Race Oncology Ltd Traitement du mélanome

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