WO2021114089A1 - Procédés d'utilisation de crocétine dans le traitement de tumeurs solides - Google Patents

Procédés d'utilisation de crocétine dans le traitement de tumeurs solides Download PDF

Info

Publication number
WO2021114089A1
WO2021114089A1 PCT/CN2019/124307 CN2019124307W WO2021114089A1 WO 2021114089 A1 WO2021114089 A1 WO 2021114089A1 CN 2019124307 W CN2019124307 W CN 2019124307W WO 2021114089 A1 WO2021114089 A1 WO 2021114089A1
Authority
WO
WIPO (PCT)
Prior art keywords
crocetin
cancer
radiotherapy
mice
anticancer
Prior art date
Application number
PCT/CN2019/124307
Other languages
English (en)
Inventor
Jiang Qian
Lang CAO
Original Assignee
Hangzhou Menglanruisi Biotechnology Co., Ltd.
Dr. Q Labs, Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Hangzhou Menglanruisi Biotechnology Co., Ltd., Dr. Q Labs, Ltd filed Critical Hangzhou Menglanruisi Biotechnology Co., Ltd.
Priority to CN201980095964.XA priority Critical patent/CN113993515B/zh
Priority to PCT/CN2019/124307 priority patent/WO2021114089A1/fr
Publication of WO2021114089A1 publication Critical patent/WO2021114089A1/fr

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/20—Carboxylic acids, e.g. valproic acid having a carboxyl group bound to a chain of seven or more carbon atoms, e.g. stearic, palmitic, arachidic acids
    • A61K31/202—Carboxylic acids, e.g. valproic acid having a carboxyl group bound to a chain of seven or more carbon atoms, e.g. stearic, palmitic, arachidic acids having three or more double bonds, e.g. linolenic
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • the present disclosure relates to crocetin that has therapeutic uses in treating solid tumors.
  • the present disclosure relates to methods of treating solid tumors, especially glioblastoma, by administering to a mammal in need thereof a pharmaceutically effective amount of crocetin, prior to, during, or after a therapy that is radiotherapy, chemotherapy, immunotherapy or a combination thereof.
  • the present disclosure also relates to the use of crocetin in the composition for treating solid tumors, especially glioblastoma.
  • hematological neoplasms and solid tumors severely threaten human health.
  • Treatments of cancers have traditionally been accomplished through one of, or a combination of, surgery, chemotherapy, radiotherapy, immunotherapy, and hormone therapy, among others.
  • new anticancer drugs especially the rise of immunotherapy, the efficacy of hematological neoplasms has recently been improved greatly, while radiotherapy and chemotherapy remain the major treatments after surgery in treatment of solid tumors.
  • radiotherapy and chemotherapy have harm or toxicity on normal tissues and organs, lead to decreased self-immunity, and can result in certain side effects, e.g., hair loss, nausea, vomit, loss of appetite, compromised immune system, myelosuppression, etc. These side effects may jeopardize the efficacy of chemotherapy and radiotherapy, and even discontinue chemotherapy and radiotherapy entirely.
  • chemotherapeutic drugs capable of targeting a specific molecule or cancer-associated signaling pathway with reduction in side effects have often failed to yield the expected improvements in patient prognoses. This is largely due to the ability of cancer cells to utilize a combination of many different cellular mechanisms to enhance viability.
  • cancer cells are able to circumvent apoptosis induced from targeted therapies by simply activating other survival pathways after the initial treatment. Meanwhile, due to the primary and acquired drug resistance of tumor cells and other factors, the therapeutic effect of chemotherapy is still unsatisfactory.
  • Glioblastoma is the most common primary intracranial tumor, caused by the canceration of brain and spinal glial cells, and characterized by high incidence, high recurrence rate, high mortality, and low cure rate. It does not significantly differentiate from normal brain tissues due to its infiltrative growth. And in most cases, it is not limited to one cerebral lobe, penetrating beyond brain tissues in fingerlike pattern and thus damaging brain tissues. Glioblastoma accounts for approximately 12%to 15%of all brain tumors (which account for 85%to 90%of all primary central nervous system (CNS) tumors) . The number of new cases for CNS tumors in the U.S. is approximately 18,800 (6.6 per 100,000 persons) per year, with around 12,800 (4.7 per 100,000 persons) deaths.
  • CNS central nervous system
  • GBM Glioblastoma multiforme
  • TMZ temozolomide
  • patients with glioblastoma receive fractionated focal irradiation in daily fractions of 2 Gy given 5 days per week for 6 weeks (total 60 Gy) plus continuous daily temozolomide 75 mg/m2/day, followed by six cycles of adjuvant temozolomide (150 to 200 mg/m 2 /day for 5 days during each 28-day cycle) after one-month break.
  • adjuvant temozolomide 150 to 200 mg/m 2 /day for 5 days during each 28-day cycle
  • patients treated by the standard treatment still have low two-year survival rate.
  • Natural products are a historically successful source of medicinally active compounds with fewer unwanted side effects, especially in regard to chemotherapeutics.
  • 63%of cancer drugs used between 1981 and 2006 were natural products, were inspired by natural products, or were synthesized from a natural pharmacophore.
  • Medicinally active compounds derived from natural materials have the potential to provide targeted cytotoxic and immune modulating responses while limiting the side effects associated with currently utilized cancer treatments.
  • the use of natural products attempts to balance a robust ability to target numerous pathways simultaneously with a historical record of safe human consumption and benign side effects.
  • Crocetin (C 20 H 24 O 4 , CAS: 27876-94-4) is a naturally-existed 8, 8′-diapo- ⁇ , ⁇ ’-carotenedioic acid with extensive physiological activities. As a brick-red crystal with a melting point of 285 °C, it is insoluble in water and most organic solvents. Crocetin has the following chemical structure:
  • crocetin As a minor compound in Saffron Yellow or Gardenia Yellow, colorants extracted from the flower of Saffron (Crocus salivus) or the fruits of Gardenia (Gardenia jasminoides) respectively, crocetin has been used for more than 1,000 years. Natural crocetin are scare in saffron and gardenia. Commercially, crocetin is mainly obtained through crocin hydrolyzation and glycosyl removal. Now it can also be obtained via chemical synthesis.
  • TSC Trans-sodium crocetinate
  • the present disclosure provides a method of treating a solid tumor in a mammal in need thereof comprising administering to the mammal a pharmaceutically effective amount of crocetin, prior to, during, or after a therapy that is radiotherapy, chemotherapy, immunotherapy or a combination thereof.
  • the mammal is human.
  • the radiotherapy is selected from the form of electromagnetic waves, such as X-rays or gamma rays, or charged particles or neutral particles.
  • the radiotherapy is administered by external beam, an interstitial implant, or a combination thereof.
  • the radiotherapy is given at a dose of about 60-70 Gy over 4-7 weeks.
  • the solid tumor disclosed herein is selected from glioblastoma, squamous cell carcinoma, skin cancer-related tumors, breast cancer, head and neck cancer, gynecological cancer, urinary and male genital cancer, bladder cancer, prostate cancer, bone cancer, endocrine adenocarcinoma, digestive tract cancer, breast cancer, major digestive/organ cancer, central nervous system cancer, and lung cancer.
  • the solid tumor is glioblastoma.
  • the chemotherapy is selected from administering the drugs with temozolomide, cisplatin, methotrexate, or paclitaxel.
  • the chemotherapy is a therapy with temozolomide.
  • the solid tumor is glioblastoma
  • the chemotherapy is a therapy with temozolomide.
  • crocetin is used as a sensitizer.
  • the method further comprises administering with at least one anticancer entity.
  • the anticancer entity disclosed herein is, for example, selected from an additional sensitizer in a cancer therapy, a targeted therapeutic agent, and an immunotherapeutic agent.
  • the anticancer entity is selected from anticancer alkylating agents, anticancer antimetabolites, anticancer antibiotics, plant-derived anticancer agents, anticancer platinum coordination compounds, anticancer camptothecin derivatives, anticancer tyrosine kinase inhibitors, monoclonal antibodies, and biological response modifiers.
  • the pharmaceutically effective amount of crocetin is administered prior to a therapy that is radiotherapy, chemotherapy, immunotherapy or a combination thereof.
  • the present disclosure provides a composition comprising crocetin and at least one pharmaceutically acceptable carrier or auxiliary for use in treatment of a solid tumor.
  • the composition is in the form of injection, tablet, capsule, pill, suppository, aerosol, oral liquid preparation, granule, powder, sustained release preparation, nano preparation, syrup, vina, tincture, lotion, film or a combination thereof.
  • the composition is in the form of liposomal formulation.
  • the composition is administered by orally, injection, implant, spray, inhalation or a combination thereof.
  • the present disclosure provides a method of sensitizing a mammal in need thereof to a therapy that is radiotherapy, chemotherapy, or a combination thereof, comprising administering to the mammal with a pharmaceutically effective amount of crocetin.
  • the mammal is human.
  • Figure 1 shows the change curves of the HCT116 subcutaneous tumor relative volume (Vt/V0) of BALB/c nude mice with the number of observation days.
  • Figure 2 shows the change curves of the Hela subcutaneous tumor relative volume (Vt/V0) of BALB/c nude mice with the number of observation days.
  • Figure 3 shows the change curves of the HepG2 subcutaneous tumor relative volume (Vt/V0) of BALB/c nude mice with the number of observation days.
  • Figure 4 shows Kaplan-Meier survival curves of BALB/c nude mice bearing orthotopic C6 glioblastoma and the statistical variance among various groups of mice.
  • Figure 5 shows Kaplan-Meier survival curves of BALB/c nude mice bearing orthotopic C6 glioblastoma and the statistical variance among various groups of mice.
  • Figure 6 shows Kaplan-Meier survival curves of BALB/c nude mice bearing orthotopic C6 glioblastoma and the statistical variance among various groups of mice.
  • Figure 7 shows Kaplan-Meier survival curves of C57BL/6 mice bearing syngeneic orthotopic GL261 glioblastoma and the statistical variance among various groups of mice.
  • Figure 8 shows Kaplan-Meier survival curves of C57BL/6 mice bearing syngeneic orthotopic GL261 glioblastoma and the statistical variance among various groups of mice.
  • A, B, C, or combinations thereof refers to all permutations and combinations of the listed items preceding the term.
  • “A, B, C, or combinations thereof” is intended to include at least one of: A, B, C, AB, AC, BC, or ABC, and if order is important in a particular context, also BA, CA, CB, CBA, BCA, ACB, BAC, or CAB.
  • expressly included are combinations that contain repeats of one or more item or term, such as BB, AAA, AB, BBC, AAABCCCC, CBBAAA, CABABB, and so forth.
  • BB BB
  • AAA AAA
  • AB BBC
  • AAABCCCCCC CBBAAA
  • CABABB CABABB
  • cancer refers to a proliferative disorder disease caused or characterized by the proliferation of cells which have lost susceptibility to normal growth control.
  • cancer includes tumors and any other proliferative disorders. Cancers of the same tissue type originate in the same tissue, and may be divided into different subtypes based on their biological characteristics.
  • the cancer may be selected, for example, from glioblastoma, squamous cell carcinoma, skin cancer-related tumors, breast cancer, head and neck cancer, gynecological cancer, urinary and male genital cancer, bladder cancer, prostate cancer, bone cancer, endocrine adenocarcinoma, digestive tract cancer, major digestive/organ cancer, central nervous system cancer, and lung cancer.
  • Glioblastoma also known as glioblastoma multiforme, may develop from a diffuse astrocytoma or an anaplastic astrocytoma but more commonly presents de novo without evidence of a less malignant precursor. Histologically, this tumor is an anaplastic, cellular glioma composed of poorly differentiated, often pleomorphic astrocytic tumor cells with marked nuclear atypia and brisk mitotic activity. Glioblastoma primarily affects the cerebral hemispheres. Central nervous system tumors are, for example, associated with characteristic patterns of altered oncogenes, altered tumor-suppressor genes, and chromosomal abnormalities.
  • Skin cancer related tumors include, for example, squamous cell carcinoma, Kaposi's sarcoma, malignant melanoma, Merkel cell skin cancer, and non-melanoma skin cancer.
  • Head-and-neck cancers include, for example, laryngeal, hypopharyngeal, nasopharyngeal, oropharyngeal cancer, lip and oral cavity cancer and squamous cell.
  • Lymphomas include, for example, AIDS-related lymphoma, non-Hodgkin's lymphoma, cutaneous T-cell lymphoma, Burkitt lymphoma, Hodgkin's disease, and lymphoma of the central nervous system.
  • breast cancer examples include, but are not limited to, invasive ductal carcinoma, invasive lobular carcinoma, ductal carcinoma in situ, and lobular carcinoma in situ.
  • cancers of the respiratory tract include, but are not limited to, small-cell and non-small-cell lung carcinoma, as well as bronchial adenoma and pleuropulmonary blastoma.
  • urinary and male genital cancers include, but are not limited to, bladder, penile, kidney, renal pelvis, ureter, urethral, testicular and human papillary renal cancers.
  • gynecological cancers include, but are not limited to, endometrial, cervical, ovarian, vaginal, and vulvar cancer, as well as sarcoma of the uterus.
  • Endocrine adenocarcinoma can be named by reference to the hormone that they produce, e.g., gastrinomas (which produce gastrin) , insulinomas (which produce insulin) , somatostatinomas (which produce somatostatin) , VIPomas (which produce VIP) and glucagonomas (which produce glucagon) .
  • gastrinomas which produce gastrin
  • insulinomas which produce insulin
  • somatostatinomas which produce somatostatin
  • VIPomas which produce VIP
  • glucagonomas which produce glucagon
  • digestive tract cancers include, but are not limited to, anal, colon, colorectal, esophageal, gallbladder, gastric, pancreatic, rectal, small-intestine, and salivary gland cancers.
  • lung cancers include, but are not limited to, small-cell lung carcinoma and non-small-cell lung carcinoma comprising squamous cell lung carcinoma, adenocarcinoma and large cell lung carcinoma.
  • the term “sensitize, ” “sensitizing, ” or “sensitizer” refers to an increased sensitivity or reduce the resistance of a cancer sample or a mammal responding to a therapeutic treatment.
  • An increased sensitivity or a reduced sensitivity to a therapeutic treatment is measured according to a known method in the art for the particular treatment and methods described herein below, including, for example, cell proliferative assays (Tanigawa N, Kern D H, Kikasa Y, Morton D L, Cancer Res, 42: 2159-2164 (1982) ) , cell death assays (Weisenthal L M, Shoemaker R H, Marsden J A, Dill P L, Baker J A, Moran E M, Cancer Res, 94: 161-173 (1984) ; Weisenthal L M, Lippman M E, Cancer Treat Rep, 69: 615-632 (1985) ; Weisenthal L M, In: Kaspers G J L, Pieters R, Twentyman P R,
  • the sensitivity or resistance may also be measured in animals by measuring the tumor size reduction over a period of time, for example, 6 months for human and 4-6 weeks for mouse.
  • a composition or a method sensitizes response to a therapeutic treatment if the increase in treatment sensitivity or the reduction in resistance is 25%or more, for example, 30%, 40%, 50%, 60%, 70%, 80%, or more, to 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, 15-fold, 20-fold or more, compared to treatment sensitivity or resistance in the absence of such composition or method.
  • the determination of sensitivity or resistance to a therapeutic treatment is routine in the art and within the skill of an ordinarily skilled clinician.
  • a “cancer therapy sensitizer” refers to a compound or a composition containing at least one compound that can sensitize the cancer therapy.
  • it refers to a compound or a composition containing an effective amount of at least one compound and a pharmaceutically acceptable carrier, diluent, excipient or a combination thereof.
  • the aforementioned compound or composition can be applied before, during or both before and during the cancer therapy to improve or enhance the effect of one or more therapeutically active compositions upon a cancer or a tumor in an individual in need, and then achieve the goal of eliminating, inhibiting, improving, comforting or preventing a cancer and its symptoms; retarding, prohibiting, reversing the rate of tumor proliferation; or the medical effects similar to the foregoing goals.
  • chemotherapy refers to the use of chemical agents to destroy cancer cells.
  • exemplary chemotherapy agents include, but are not limited to, actinomycin D, adriamycin, altretamine, asparaginase, bleomycin, busulphan, capecitabine, carboplatin, carmustine, chlorambucil, cisplatin, CPT-11, cyclophosphamide, cytarabine, dacarbazine, daunorubicin, doxorubicin, epirubicin, etoposide, fludarabine, fluorouracil, gemcitabine, hydroxyurea, idarubicin, fosfamide, irinotecan, liposomal doxorubicin, lomustine, melphalan, mercaptopurine, methotrexate, mitomycin, mitozantrone, oxaliplatin, procarbazine, steroids, streptozocin, taxol
  • Radiotherapy also called “radiation therapy, ” refers to the treatment of cancer and other diseases with ionizing radiation. Ionizing radiation deposits energy that injures or destroys cells in the area being treated (i.e., the “target tissue” ) by damaging their genetic material, making it impossible for these cells to continue to grow. Although radiation damages both cancer and normal cells, the normal cells are able to repair themselves and function properly. Radiotherapy may be used to treat localized solid tumors, such as cancers of the skin, tongue, larynx, brain, breast, or uterine cervix. It can also be used to treat leukemia and lymphoma (cancers of the blood-forming cells and lymphatic system, respectively) .
  • Exemplary radiotherapy may be selected from the forms of electromagnetic waves, such as X-rays or gamma rays, or charged particles or neutral particles.
  • the radiotherapy may be administered by external beam, an interstitial implant, or a combination thereof.
  • a course of radiotherapy consisting of 60-70 Gy for the majority of tumors over 4-7 weeks.
  • the radiation and chemotherapy sensitizer disclosed herein is a pharmaceutical compound or composition that can be used prior to, or simultaneously with, radiotherapy as well as chemotherapy, to strengthen the effect that radiotherapy and chemotherapy has on the tumor.
  • administer refers to introduce by any means a compound or composition (e.g., a therapeutic agent) into the body of a mammal in order to prevent or treat a disease or condition (e.g., cancer) .
  • a disease or condition e.g., cancer
  • treating, ” “treatment, ” “therapy, ” and “therapeutic treatment” as used herein refer to curative therapy, prophylactic therapy, or preventative therapy. These terms also describe the management and care of a mammal for the purpose of combating a disease, or related condition, and include the administration of a composition to alleviate the symptoms, side effects, or other complications of the disease or condition.
  • Therapeutic treatment for cancer includes, for example, surgery, chemotherapy, radiation therapy, gene therapy, and immunotherapy.
  • the term “mammal” refers to human or other animals, such as farm animals or laboratory animals (e.g. guinea pig or mice) .
  • the mammal is human. It may be a human who has been diagnosed as in need of treatment for a disease or disorder disclosed herein.
  • “Pharmaceutically effective amount” encompasses an amount sufficient to ameliorate or prevent a symptom or sign of the medical condition.
  • An effective amount for a particular patient or a veterinary subject may vary depending on factors, such as the condition being treated, the overall health of the patient, the method route and dose of administration and the severity of side effects.
  • a pharmaceutically effective amount can be the maximal dose or dosing protocol that avoids significant side effects or toxic effects.
  • a diagnostic measure or parameter by at least 5%, such as by at least 10%, further such as at least 20%, further such as at least 30%, further such as at least 40%, further such as at least 50%, further such as at least 60%, further such as at least 70%, further such as at least 80%, and even further such as at least 90%, wherein 100%is defined as the diagnostic parameter shown by a normal subject.
  • a pharmaceutically effective amount of crocetin would be an amount that is, for example, sufficient to reduce a tumor volume, inhibit tumor growth, or prevent or reduce metastasis, prior to, during, or after a cancer therapy.
  • pharmaceutically acceptable refers to those compounds, materials, compositions, and/or dosage forms, which are suitable for use in contact with the tissues of human beings or animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.
  • compositions disclosed herein may be tableted with conventional tablet bases such as lactose, sucrose and cornstarch in combination with binders such as acacia, corn starch or gelatin, disintegrating agents intended to assist the break-up and dissolution of the tablet following administration such as potato starch, alginic acid, corn starch, and guar gum, gum tragacanth, acacia, lubricants intended to improve the flow of tablet granulation and to prevent the adhesion of tablet material to the surfaces of the tablet dies and punches, for example, talc, stearic acid, or magnesium, calcium or zinc stearate, dyes, coloring agents, and flavoring agents such as peppermint, oil of wintergreen, or cherry flavoring, intended to enhance the aesthetic qualities of the tablets and make them more acceptable to the patient.
  • binders such as acacia, corn starch or gelatin
  • disintegrating agents intended to assist the break-up and dissolution of the tablet following administration such as potato starch, alginic acid, corn
  • Suitable excipients for use in oral liquid dosage forms include, for example, dicalcium phosphate and diluents such as water and alcohols, for example, ethanol, benzyl alcohol, and polyethylene alcohols, either with or without the addition of a pharmaceutically acceptable surfactant, suspending agent or emulsifying agent.
  • Various other materials may be present as coatings or to otherwise modify the physical form of the dosage unit. For instance, tablets, pills or capsules may be coated with shellac, sugar or both.
  • Dispersible powders and granules are suitable for the preparation of an aqueous suspension. They provide the active ingredient in admixture with a dispersing or wetting agent, a suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are exemplified by those already mentioned above. Additional excipients, for example, those sweetening, flavoring and coloring agents described above, may also be present.
  • the pharmaceutical compositions disclosed herein may also be in the form of oil-in-water emulsions.
  • the oily phase may be a vegetable oil such as liquid paraffin or a mixture of vegetable oils.
  • Suitable emulsifying agents may, for example, be chosen from (1) naturally occurring gums, such as gum acacia and gum tragacanth, (2) naturally occurring phosphatides, such as soy bean and lecithin, (3) esters or partial esters derived from fatty acids and hexitol anhydrides, for example, sorbitan monooleate, (4) condensation products of the partial esters with ethylene oxide, for example, polyoxyethylene sorbitan monooleate.
  • the emulsions may also contain, for example, sweetening and flavoring agents.
  • Oily suspensions may be formulated by suspending the active ingredient in a vegetable oil, such as arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin.
  • the oily suspensions may contain a thickening agent, such as beeswax, hard paraffin, or cetyl alcohol.
  • the suspensions may also contain one or more preservatives, for example, ethyl or n-propyl p-hydroxybenzoate; one or more coloring agents; one or more flavoring agents; and one or more sweetening agents such as sucrose or saccharin.
  • Syrups and elixirs may be formulated with sweetening agents, such as glycerol, propylene glycol, sorbitol or sucrose.
  • sweetening agents such as glycerol, propylene glycol, sorbitol or sucrose.
  • Such formulations may also contain, for example, at least one entity chosen from demulcents, preservatives, such as methyl and propyl parabens, and flavoring and coloring agents.
  • the combinations disclosed herein may also be administered parenterally, that is, subcutaneously, intravenously, intraocularly, intramuscularly, or interperitoneally, as injectable dosages of the crocetin in, for example, a physiologically acceptable diluent with a pharmaceutical carrier that can be a sterile liquid or mixture of liquids such as water, saline, aqueous dextrose and related sugar solutions, an alcohol such as ethanol, isopropanol, or hexadecyl alcohol, glycols such as propylene glycol or polyethylene glycol, glycerol ketals such as 2, 2-dimethyl-1, 1-dioxolane-4-methanol, ethers such as polyethylene glycol) 400, an oil, a fatty acid, a fatty acid ester or, a fatty acid glyceride, or an acetylated fatty acid glyceride, with or without the addition of a pharmaceutically acceptable surfactant such as a soap
  • Suitable fatty acids include oleic acid, stearic acid, isostearic acid, and myristic acid.
  • Suitable fatty acid esters are, for example, ethyl oleate and isopropyl myristate.
  • Suitable soaps include fatty acid alkali metal, ammonium, and triethanolamine salts and suitable detergents include cationic detergents, for example dimethyl dialkyl ammonium halides, aIkyI pyridinium halides, and alkylamine acetates; anionic detergents, for example, alkyl, aryl, and olefin sulfonates, alkyl, olefin, ether, and monoglyceride sulfates, and sulfosuccinates; non-ionic detergents, for example, fatty amine oxides, fatty acid alkanolamides, and poly (oxyethylene-oxypropylene) sor ethylene oxide or propylene oxide copolymers; and amphoteric detergents, for example, alkyl-beta-aminopropionates, and 2-alkylimidazoline quaternary ammonium salts, as well as mixtures.
  • suitable detergents include cationic detergents,
  • Illustrative of the surfactants used in parenteral formulations are the class of polyethylene sorbitan fatty acid esters, for example, sorbitan monooleate and the high molecular weight adducts of ethylene oxide with a hydrophobic base, formed by the condensation of propylene oxide with propylene glycol.
  • compositions disclosed herein may be in the form of sterile injectable aqueous suspensions.
  • suspensions may be formulated according to known methods using suitable dispersing or wetting agents and suspending agents, such as, sodium carboxymethylcellulose, methylcellulose, hydroxypropyl methyl cellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia; dispersing or wetting agents that may be a naturally occurring phosphatide such as lecithin, a condensation product of an alkylene oxide with a fatty acid, for example, polyoxyethylene stearate, a condensation product of ethylene oxide with a long chain aliphatic alcohol, for example, heptadeca-ethyleneoxycetanol, a condensation product of ethylene oxide with a partial ester derived from a fatty acid and a hexitol such as polyoxyethylene sorbitol monooleate, or a condensation product of an ethylene oxide with a partial ester derived from
  • the sterile injectable preparation disclosed herein may also be, for example, a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent.
  • Diluents and solvents that may be employed are, for example, water, Ringer's solution, isotonic sodium chloride solutions and isotonic glucose solutions.
  • sterile fixed oils are conventionally employed as solvents or suspending media.
  • any bland, fixed oil may be employed including synthetic mono-or diglycerides.
  • fatty acids such as oleic acid can be used in the preparation of injectables.
  • compositions disclosed herein may also be administered in the form of suppositories for rectal administration of the drug.
  • These compositions can be prepared, for example, by mixing the drug with a suitable non-irritation excipient that is solid at the room temperature but liquid at the rectal temperature and will therefore melt in the rectum to release the drug.
  • a suitable non-irritation excipient that is solid at the room temperature but liquid at the rectal temperature and will therefore melt in the rectum to release the drug.
  • Such materials are chosen, for example, from cocoa butter and polyethylene glycol.
  • Controlled release formulations for parenteral administration include, for example, liposomal, polymeric microsphere and polymeric gel formulations that are known in the art.
  • compositions disclosed herein can also contain other conventional pharmaceutically acceptable ingredients, generally referred to as carriers, diluents, or auxiliaries, as necessary or desired.
  • Conventional procedures for preparing such compositions in appropriate dosage forms can be utilized. Such ingredients and procedures include those described in the following references: Powell, M.F. et al, "Compendium of Excipients for Parenteral Formulations” , PDA Journal of Pharmaceutical Science ft Technology 52 (5) , 238-311 (1998) ; Strickley, R. G "Parenteral Formulations of Small Molecule Therapeutics Marketed in the United States (1999) -Part-1" PDA Journal of Pharmaceutical Science & Technology 53 (6) , 324-349 (1999) ; and Nema, S.
  • compositions disclosed herein for its intended route of administration include, for example, acidifying agents (examples include, but are not limited to, acetic acid, citric acid, fumaric acid, hydrochloric acid, and nitric acid) ; alkalinizing agents (examples include, but are not limited to, ammonia solution, ammonium carbonate, diethanolamine, monoethanolamine, potassium hydroxide, sodium borate, sodium carbonate, sodium hydroxide, and triethanolamine, trolamine) ; adsorbents (examples include, but are not limited to, powdered cellulose and activated charcoal) ; aerosol propellants (examples include, but are not limited to, carbon dioxide, chlorofluorocarbon such as Freon-11 (CCl 3 F) , Freon
  • clarifying agents include, but are not limited to, bentonite
  • emulsifying agents include, but are not limited to, acacia, cetomacrogol, cetyl alcohol, glyceryl monostearate, lecithin, sorbitan monooleate, and polyoxyethylene 50 monostearate
  • encapsulating agents include, but are not limited to, gelatin and cellulose acetate phthalate
  • flavorants include, but are not limited to, anise oil, cinnamon oil, cocoa, menthol, orange oil, peppermint oil, and vanillin
  • humectants include, but are not limited to, glycerol, propylene
  • the injection can be directly intratumorally injected using, for example, a syringe, or indirectly injected via, for example, an angiography catheter into the tumor area at time of the irradiation.
  • the sensitizer liquid or gel disclosed herein can be injected into the target tumor area using, for example, a syringe or angiography catheter, before, after or at the same time as the administration of the anti-cancer agent.
  • a syringe having a needle of about 21 gauge can be used to perform the intratumoral injection guided by ultrasonographic examination while observing the state of permeation of the sensitizer into the tissue.
  • the sensitizer can be delivered widely to the tissue under ultrasonographic guidance to modify the depth and direction of the injection needle.
  • the dose of sensitizer administered to the tumor area differs depending on the size of the tumor and the administration method.
  • Anti-cancer agents used herein include, for example, alkylating agents such as cyclophosphamide, ifosfamide, busulfan, melphalan, bendamustine hydrochloride, nimustine hydrochloride, ranimustine, dacarbazine, procarbazine hydrochloride, and temozolomide; antimetabolites such as methotrexate, pemetrexed sodium, fluorouracil, doxifluridine, capecitabine, tegafur, cytarabine, cytarabine ocfosfate hydrate, enocitabine, gemcitabine hydrochloride, mercaptopurine hydrate, fludarabine phosphate, nelarabine, pentostatin, cladribine, levofolinate calcium, calcium folinate, hydroxycarbamide, L-asparaginase, and azacitidine; antitumor antibiotics such as doxorubi
  • anti-cancer agents examples include anthracycline anti-cancer agents such as doxorubicin hydrochloride, daunorubicin hydrochloride, pirarubicin, epirubicin hydrochloride, idarubicin hydrochloride, aclarubicin hydrochloride, amrubicin hydrochloride, and mitoxantrone hydrochloride; platinum anti-cancer agents such as cisplatin, miriplatin hydrate, carboplatin, nedaplatin, and oxaliplatin; and pyrimidine antimetabolite-based anti-cancer agents such as fluorouracil, doxifluridine, capecitabine, tegafur, cytarabine, cytarabine ocfostate hydrate, enocitabine, and gemcitabine hydrochloride.
  • Crocetin can successfully be employed as, for example, a chemotherapy and/or radiotherapy sensitizer, for various solid tumors that were grafted onto mice. It can synergize with radiotherapy and/or chemotherapy to prolong the survival of tumor-bearing mice.
  • crocetin can have the following advantages: (1) the overall survival of tumor-bearing mice treated with crocetin in combination with chemo-radiation therapy can be significantly longer than those treated with TSC in combination with chemo-radiation therapy at the same dose; (2) crocetin can not only enhance the sensitivity of tumor cells to radiotherapy, but also enhance the sensitivity of tumor cells to chemotherapy; (3) the optimal dose of TSC achieving the maximum synergistic effect with chemo-radiation therapy is 100 ⁇ g/kg, followed by a dose-dependent decrease, while the synergistic effect of crocetin with chemo-radiation therapy can be dose-dependently increasing in the range of 100-400 ⁇ g/kg.
  • Crocetin enhances radiotherapy to suppress the growth rates of subcutaneous solid tumors in BALB/c nude mice
  • Hela human cervical cancer cell line HCT116 human colonic cancer cell line
  • HepG2 human hepatic carcinoma cell line ATCC
  • Crocetin brick-red powder, HPLC purity greater than 98% (Shaoxing Tiankang Biotechnology Co., Ltd. ) .
  • Intravenous injection of crocetin the liposome preparation was prepared according to Example 1 and diluted to certain concentration with saline; intragastric administration of crocetin: a crocetin suspension with certain concentration was formulated by use of 0.5%sodium carboxymethyl cellulose.
  • TSC was prepared from crocetin according to the method disclosed in U.S. Patent No. 6,060,511, with an HPLC purity of greater than 98%.
  • mice of 4-8 weeks in age Male BALB/c nude mice of 4-8 weeks in age, weighing about 18 ⁇ 2 g, were provided by, and maintained in a SPF facility under sterile atmosphere at the animal facility of the Center of Laboratory Animals, Zhejiang Academy of Medical Sciences. 2 ⁇ 10 6 Hela, HCT116, and HepG2 cells were re-suspended in 200 ⁇ l PBS and injected subcutaneously into the right flank of the mice, respectively. The inoculated sites and the number of tumor cells inoculated were the same for each mouse, and each cell line in four mice (12 in total) .
  • tumor tissues with good growth (no degeneration or necrosis) and reddish color were selected for next inoculation, and the inoculated tumor tissue was about 2 mm in diameter for each mouse. Each type of tumor tissue was inoculated into 25 mice.
  • mice bearing same type of tumor were randomly assigned to one of five groups:
  • Control group (Control) ;
  • Radiotherapy + crocetin i.v. group (RT + crocetin i.v. (100 ⁇ g/kg) ) ;
  • Radiotherapy + crocetin i.g. group (RT + crocetin i.g. (200 ⁇ g/kg) ) .
  • mice All mice were treated for consecutive five days. Each mouse in groups 2-5 received a single dose (5 Gy) of local irradiation (RS-2000-PRO-225 (RAD SOURCE, Simmens) ) within 30 minutes after administration of test compounds on day 3 ( Figure 1, 2 and 3) .
  • RS-2000-PRO-225 RAD SOURCE, Simmens
  • Tumor volumes were measured using caliper two or three times a week until 4-fold of the volume at the beginning of treatment.
  • the relative tumor volume-time curve was plotted.
  • the statistical difference between different groups was analyzed by one-way ANOVA.
  • a P ⁇ 0.05 was considered to be statistically significant.
  • Figure 1 shows the inhibitory effects of the treatment of crocetin or TSC together with radiation against HCT116 subcutaneous tumor in mice.
  • Figure 2 shows the inhibitory effects of the treatment of crocetin or TSC together with radiation against Hela subcutaneous tumor in mice.
  • Figure 3 shows the inhibitory effects of the treatment of crocetin or TSC together with radiation against HepG2 subcutaneous tumor in mice.
  • all other groups could effectively inhibit the growth of subcutaneous tumors (p ⁇ 0.05) .
  • TSC or crocetin together with radiation group significantly delay the growth of subcutaneous tumors (P ⁇ 0.01) .
  • crocetin in the present disclosure could significantly enhance radiotherapy to suppress the growth of a plurality of solid tumors.
  • C6 glioma cell line (ATCC) .
  • Crocetin brick-red powder, HPLC purity greater than 98% (Shaoxing Tiankang Biotechnology Co., Ltd. ) .
  • Intravenous injection of crocetin the liposome preparation was prepared according to Example 1 and diluted to certain concentration with saline; intragastric administration of crocetin: a crocetin suspension with certain concentration was formulated by use of 0.5%sodium carboxymethyl cellulose.
  • TSC was prepared from crocetin according to the method disclosed in U.S. Patent No. 6,060,511, with an HPLC purity of greater than 98%.
  • Temozolomide has a purity greater than 99% (from Guangzhou Tomums Life Science Co., Ltd. ) .
  • mice of 6-8 weeks in age Male BALB/c nude mice of 6-8 weeks in age, weighing about 18 ⁇ 2 g, were provided by, and maintained in a SPF facility under sterile atmosphere at the animal facility of the Center of Laboratory Animals, Zhejiang Academy of Medical Sciences.
  • C6 glioma cells were prepared for suspensions concentrated at 2 ⁇ 10 6 cells/10 ⁇ l in PBS.
  • Each mouse was intracranially inoculated with 2.5 ⁇ l of cell suspension (5 ⁇ 10 5 cells) .
  • the inoculated sites and the number of tumor cells inoculated were the same for each mouse, and a total of 50 mice were inoculated.
  • mice After inoculation for 7 days, 50 mice were randomly assigned to one of the six groups, and each group has 8-9 mice;
  • Control group (Control) ;
  • TSC i.v. group (TSC (100 ⁇ g/kg) ) ;
  • Crocetin i.v. group Crocetin (100 ⁇ g/kg) ) .
  • mice were intravenously injected with TSC, crocetin or saline (control) for consecutive 5 days, followed by intraperitoneal administration of temozolomide (50 mg/kg, 149 mg/m 2 ) 20 minutes later.
  • mice were examined daily for alertness, feeding ability, external appearance, focal motor deficits, fecal traits and response to external stimuli.
  • the body weight of each mouse during the survival was continuously weighed, and the survival duration of each mouse was recorded.
  • the survival rate of each group was calculated by formula: number of mice remaining in each group /total number of mice in each group at the beginning.
  • the survival curve was plotted using Kaplan-Meier methodology.
  • Log-Rank test Mantel-Cox was used to determine the difference between the survival distributions of different groups.
  • a P ⁇ 0.05 was considered to be statistically significant.
  • Figure 4 shows the Kaplan-Meier survival curves of mice and the statistical variance among survival curves of mice in various groups.
  • Treatment with TSC or crocetin alone could not prolong the survival relative to untreated mice.
  • Treatment with TMZ improved overall survival relative to untreated mice, adding 4 days to the median survival time (25 vs. 21 days, P ⁇ 0.01) .
  • Crocetin could further enhance such effect of TMZ, adding another 4 days to the median survival time (29 vs. 25 days, P ⁇ 0.01) , while TSC did not have such effect.
  • crocetin in the present disclosure could significantly enhance the inhibitory effects of TMZ against the C6 orthotopic glioblastoma in BALB/c nude mice, therefore extend the survival of orthotopic tumor-planted mice.
  • Crocetin synergizes with chemo-radiation therapy to enhance survival in BALB/c nude mice bearing orthotopic C6 glioblastoma
  • C6 glioma cell line (ATCC) .
  • Crocetin brick-red powder, HPLC purity greater than 98% (Shaoxing Tiankang Biotechnology Co., Ltd. ) .
  • Intravenous injection of crocetin the liposome preparation was prepared according to Example 1 and diluted to certain concentration with saline; intragastric administration of crocetin: a crocetin suspension with certain concentration was formulated by use of 0.5%sodium carboxymethyl cellulose.
  • TSC was prepared from crocetin according to the method disclosed in U.S. Patent No. 6,060,511, with an HPLC purity of greater than 98%.
  • Temozolomide has a purity greater than 99% (from Guangzhou Tomums Life Science Co., Ltd. ) .
  • mice of 4-8 weeks in age Male BALB/c nude mice of 4-8 weeks in age, weighing about 18 ⁇ 2 g, were provided by, and maintained in a SPF facility under sterile atmosphere at the animal facility of the Center of Laboratory Animals, Zhejiang Academy of Medical Sciences.
  • C6 glioma cells were prepared for suspensions concentrated at 2 ⁇ 10 6 cells/10 ⁇ l in PBS.
  • Each mouse was intracranially inoculated with 2.5 ⁇ l of cell suspension (5 ⁇ 10 5 cells) .
  • the inoculated sites and the number of tumor cells inoculated were the same for each mouse, and a total of 50 mice were inoculated.
  • mice After inoculation for 7 days, 50 mice were randomly assigned to one of the six groups, and each group has 8-9 mice;
  • Control group (Control) ;
  • TMZ + radiotherapy + crocetin i.v. group (TMZ + RT + TK10 i.v. (100 ⁇ g/kg) ) ;
  • TMZ + radiotherapy + crocetin i.v. group (TMZ + RT + TK10 i.v. (200 ⁇ g/kg) ) ;
  • TMZ + radiotherapy + crocetin i.g. group (TMZ + RT + TK10 i.g. (200 ⁇ g/kg) .
  • mice From Days 8 to 12, all mice were intravenously injected with TSC, crocetin or saline (control) for consecutive 5 days, followed by intraperitoneal administration of temozolomide (50 mg/kg, 149 mg/m 2 ) 20 minutes later.
  • mice in Groups 2-6 received 8 Gy local irradiation (RS-2000-PRO-225 (RAD SOURCE) ) within 30 minutes after administration of test compounds on Day 12
  • mice were examined daily for alertness, feeding ability, external appearance, focal motor deficits, fecal traits and response to external stimuli.
  • the body weight of each mouse during the survival was continuously weighed, and the survival duration of each mouse was recorded.
  • the survival rate of each group was calculated by formula: number of mice remaining in each group /total number of mice in each group at the beginning.
  • the survival curve was plotted using Kaplan-Meier methodology.
  • Log-Rank test Mantel-Cox was used to determine the difference between the survival distributions of different groups.
  • a P ⁇ 0.05 was considered to be statistically significant.
  • Figure 5 shows the survival curves of each group of mice changing with the number of observation days, and the statistical variance among the survival curves of various groups of mice.
  • Treatment with TMZ + Radiation improved overall survival relative to untreated mice, adding 7 days to the median survival time (31 vs. 24 days, P ⁇ 0.01) .
  • Intravenous injection or intragastric administration of crocetin, as well as Intravenous injection of TSC could further enhance such effect of TMZ, extend 6-8 days to the median survival time, respectively (39, 38, 37 vs. 31 days, respectively, P ⁇ 0.01) .
  • crocetin in the present disclosure could significantly enhance the inhibitory effects of chemo-radiation therapy against the C6 orthotopic glioblastoma, therefore extend the survival of orthotopic tumor-bearing mice.
  • Crocetin synergizes with chemo-radiation therapy plus chemotherapy to enhance survival in BALB/c nude mice with orthotopic C6 glioblastoma
  • C6 glioma cell line (ATCC) .
  • Crocetin brick-red powder, HPLC purity greater than 98% (Shaoxing Tiankang Biotechnology Co., Ltd. ) .
  • Intravenous injection of crocetin the liposome preparation was prepared according to Example 1 and diluted to certain concentration with saline;
  • Intragastric administration of crocetin a crocetin suspension with certain concentration was formulated by use of 0.5%sodium carboxymethyl cellulose.
  • TSC was prepared from crocetin according to the method disclosed in U.S. Patent No. 6,060,511, with an HPLC purity of greater than 98%.
  • Temozolomide (TMZ) , with purity greater than 99% (from Guangzhou Tomums Life Science Co., Ltd. ) .
  • mice of 6-8 weeks in age Male BALB/c nude mice of 6-8 weeks in age, weighing about 18 ⁇ 2 g, were provided by, and maintained in a SPF facility under sterile atmosphere at the animal facility of the Center of Laboratory Animals, Zhejiang Academy of Medical Sciences.
  • C6 glioma cells were prepared for suspensions concentrated at 2 ⁇ 10 6 cells/10 ⁇ l in PBS. Each mouse was intracranially inoculated with 2.5 ⁇ l of cell suspension (5 ⁇ 10 5 cells) .
  • mice After inoculation for 6 days, 41 mice were randomly assigned to one of five groups, and each group has 8-9 mice;
  • TMZ + radiotherapy + crocetin i.v. group (TMZ + RT + TK10 i.v. (100 ⁇ g/kg) ) ;
  • TMZ + radiotherapy + crocetin i.g. group (TMZ + RT + TK10 i.g. (200 ⁇ g/kg) ) .
  • mice bearing tumors were treated with two phases: chemo-radiation therapy and chemotherapy.
  • mice were intravenously (or intragastrically) injected with TSC, crocetin or saline (control) for consecutive 5 days, followed by intraperitoneal administration of temozolomide (50 mg/kg, 149 mg/m 2 ) 20 minutes later.
  • mice in groups 2-5 received 8 Gy local irradiation (RS-2000-PRO-225 (RAD SOURCE) ) within 30 minutes after dosing on Day 11.
  • mice were intravenously (or intragastrically) injected with TSC, crocetin or saline (control) for consecutive 5 days, followed by intraperitoneal administration of temozolomide (50 mg/kg, 149 mg/m 2 ) 20 minutes later.
  • mice were examined daily for alertness, feeding ability, external appearance, focal motor deficits, fecal traits and response to external stimuli.
  • the body weight of each mouse during the survival was continuously weighed, and the survival duration of each mouse was recorded.
  • the survival rate of each group was calculated by formula: number of mice remaining in each group /total number of mice in each group at the beginning.
  • the survival curve was plotted using Kaplan-Meier methodology.
  • Log-Rank test Mantel-Cox was used to determine the difference between the survival distributions of different groups.
  • a P ⁇ 0.05 was considered to be statistically significant.
  • Figure 6 shows the survival curves of each group of mice changing with the number of observation days, and the statistical variance among the survival curves of various groups of mice.
  • Treatment with chemo-radiation therapy plus chemotherapy improved overall survival relative to untreated mice, adding 14 days to the median survival time (36 vs. 22 days, P ⁇ 0.01) .
  • Intravenous injection of TSC could enhance such effect of chemo-radiation therapy plus chemotherapy, extend 5 days to the median survival time (41 vs. 36, P ⁇ 0.01) .
  • Intravenous injection or intragastric administration of crocetin could further enhance the effect of chemo-radiation therapy plus chemotherapy, extend 9 days to the median survival time (45 vs. 36 days, respectively, P ⁇ 0.01) .
  • mice treated with crocetin +TMZ + radiation there are statistically significant difference (45 vs. 41 days, p ⁇ 0.01) of overall survival between mice treated with crocetin +TMZ + radiation and mice treated with TSC + TMZ + radiation (45 vs. 41 days, p ⁇ 0.01) .
  • crocetin in the present disclosure could significantly enhance the inhibitory effects of chemo-radiation therapy plus chemotherapy against the C6 orthotopic glioblastoma, therefore extend the survival of orthotopic tumor-bearing mice.
  • GL261 glioblastoma cell line ATCC
  • Crocetin brick-red powder, HPLC purity greater than 98% (Shaoxing Tiankang Biotechnology Co., Ltd. ) .
  • Intravenous injection of crocetin the liposome preparation was prepared according to Example 1 and diluted to certain concentration with saline;
  • Intragastric administration of crocetin a crocetin suspension with certain concentration was formulated by use of 0.5%sodium carboxymethyl cellulose.
  • TSC was prepared from crocetin according to the method disclosed in U.S. Patent No. 6,060,511, with an HPLC purity of greater than 98%.
  • Temozolomide has a purity greater than 99% (from Guangzhou Tomums Life Science Co., Ltd. ) .
  • mice of 4-8 weeks in age Male C57BL/6 mice of 4-8 weeks in age, weighing about 18 ⁇ 2 g, were provided by, and maintained in a SPF facility under sterile atmosphere at the animal facility of the Center of Laboratory Animals, Zhejiang Academy of Medical Sciences.
  • GL261 glioblastoma cells were prepared for suspensions concentrated at 2 ⁇ 10 6 cells/10 ⁇ l in PBS.
  • Each mouse was intracranially inoculated with 2.5 ⁇ l of cell suspension (5 ⁇ 10 5 cells) .
  • tumor-bearing mice were randomly assigned to one of six groups, and each group has 8-9 mice;
  • Control group (Control) ;
  • TMZ + radiotherapy + crocetin i.v. group (TMZ + RT + TK10 i.v. (100 ⁇ g/kg) ) ;
  • TMZ + radiotherapy + crocetin i.v. group (TMZ + RT + TK10 i.v. (300 ⁇ g/kg) ) ;
  • TMZ + radiotherapy + crocetin i.g. group (TMZ + RT + TK10 i.g. (200 ⁇ g/kg) ) .
  • mice were intravenously or intragastically injected with TSC, crocetin or physiological saline (control) for consecutive 3 days, followed by single intraperitoneal administration of temozolomide (100 mg/kg, 298 mg/m 2 ) 20 minutes later on day 10.
  • mice in Groups 2-6 received 5 Gy local irradiation (RS-2000-PRO-225 (RAD SOURCE) ) within 30 minutes after administration of test compounds on Day 11
  • mice were examined daily for alertness, feeding ability, external appearance, focal motor deficits, fecal traits and response to external stimuli.
  • the body weight of each mice during the survival was continuously weighed, and the survival duration of each mouse was recorded.
  • the survival rate of each group was calculated by formula: number of mice remaining in each group /total number of mice in each group at the beginning.
  • the survival curve was plotted using Kaplan-Meier methodology.
  • Log-Rank test Mantel-Cox was used to determine the difference between the survival distributions of different groups.
  • a P ⁇ 0.05 was considered to be statistically significant.
  • Figure 7 shows the Kaplan-Meier survival curves of mice and the statistical variance among survival curves of mice in various groups.
  • the other treatment groups could all effectively extend the survival of tumor-bearing mice (P ⁇ 0.05) , with their median survival increasing from 25 days to over 36 days.
  • the efficacy of TMZ + RT combining with TSC or crocetin improved (P ⁇ 0.01) , with their median survival time increasing from 36 days to over 45 days.
  • TSC + TMZ + RT group the efficacy of crocetin + TMZ + RT group (i.g. or i.v.
  • mice treated by crocetin + TMZ + RT had a statistically significant survival advantage over mice in the other treatment groups (P ⁇ 0.01) .
  • crocetin in the present disclosure could significantly enhance the inhibitory effects of chemo-radiation therapy against the syngeneic orthotopic GL261 glioblastoma model of C57BL/6 mice, therefore to extend the survival of tumor-bearing mice.
  • GL261 glioblastoma cell line ATCC
  • Crocetin brick-red powder, HPLC purity greater than 98% (Shaoxing Tiankang Biotechnology Co., Ltd. ) .
  • Intravenous injection of crocetin the liposome preparation was prepared according to Example 1 and diluted to certain concentration with saline;
  • Intragastric administration of crocetin a crocetin suspension with certain concentration was formulated by use of 0.5%sodium carboxymethyl cellulose.
  • TSC was prepared from crocetin according to the method disclosed in U.S. Patent No. 6,060,511, with an HPLC purity of greater than 98%.
  • Temozolomide has a purity greater than 99% (from Guangzhou Tomums Life Science Co., Ltd. ) .
  • mice of 6-8 weeks in age Male C57BL/6 mice of 6-8 weeks in age, weighing about 18 ⁇ 2 g, were provided by, and maintained in a SPF facility under sterile atmosphere at the animal facility of the Center of Laboratory Animals, Zhejiang Academy of Medical Sciences.
  • GL261 glioblastoma cells were prepared for suspensions concentrated at 2 ⁇ 10 6 cells/10 ⁇ l in PBS.
  • Each mouse was intracranially inoculated with 2.5 ⁇ l of cell suspension (5 ⁇ 10 5 cells) .
  • mice After inoculation for 11 days, 42 tumor-bearing mice were randomly assigned to one of five groups, and each group has 8-9 mice;
  • Control group (Control) ;
  • TMZ + radiotherapy + crocetin i.v. group (TMZ + RT + crocetin i.v. (400 ⁇ g/kg) ) ;
  • mice were administered for three consecutive days. They were intravenously injected with crocetin or saline (control) on Day 11. On Day 12, mice were intravenously injected with crocetin or saline, followed by single intraperitoneal administration of temozolomide (100 mg/kg, 298 mg/m 2 ) 20 minutes later. On Day 13, mice in groups 2-5 were intravenously injected with crocetin or saline first, and received 5 Gy local irradiation (RS-2000-PRO-225 (RAD SOURCE) ) within 30 minutes.
  • RS-2000-PRO-225 RAD SOURCE
  • mice were examined daily for alertness, feeding ability, external appearance, focal motor deficits, fecal traits and response to external stimuli.
  • the body weight of each mouse during the survival was continuously weighed, and the survival duration of each mouse was recorded.
  • the survival rate of each group was calculated by formula: number of mice remaining in each group /total number of mice in each group at the beginning.
  • the survival curve was plotted using Kaplan-Meier methodology.
  • Log-Rank test Mantel-Cox was used to determine the difference between the survival distributions of different groups.
  • a P ⁇ 0.05 was considered to be statistically significant.
  • Figure 8 shows the Kaplan-Meier survival curves of mice and the statistical variance among survival curves of mice in various groups.
  • other treatment groups could all effectively extend the survival time of tumor-bearing mice (P ⁇ 0.01) , with their median survival period increased from 24 days to over 35 days.
  • the efficacy of TMZ + RT combining with different c doses of crocetin improved (P ⁇ 0.01) , with their median survival period increasing from 35 days to over 44 days.
  • crocetin in the present disclosure can significantly enhance the inhibitory effects of chemo-radiation therapy against the syngeneic orthotopic GL261 glioblastoma model of C57BL/6 mice, therefore to extend the survival of tumor-bearing mice in a dose-dependent manner.

Landscapes

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

Abstract

La présente invention concerne la nouvelle utilisation de crocétine. En particulier, elle concerne des procédés de traitement de tumeurs solides, telles qu'un glioblastome, par administration, à un mammifère en ayant besoin, d'une quantité pharmaceutiquement efficace de crocétine, avant, pendant, ou après une thérapie qui est une radiothérapie, une chimiothérapie, une immunothérapie ou une combinaison de celles-ci. L'invention concerne également l'utilisation de crocétine dans une composition pour le traitement de tumeurs solides, telles qu'un glioblastome.
PCT/CN2019/124307 2019-12-10 2019-12-10 Procédés d'utilisation de crocétine dans le traitement de tumeurs solides WO2021114089A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
CN201980095964.XA CN113993515B (zh) 2019-12-10 2019-12-10 使用藏红花酸治疗实体肿瘤的方法
PCT/CN2019/124307 WO2021114089A1 (fr) 2019-12-10 2019-12-10 Procédés d'utilisation de crocétine dans le traitement de tumeurs solides

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/CN2019/124307 WO2021114089A1 (fr) 2019-12-10 2019-12-10 Procédés d'utilisation de crocétine dans le traitement de tumeurs solides

Publications (1)

Publication Number Publication Date
WO2021114089A1 true WO2021114089A1 (fr) 2021-06-17

Family

ID=76329129

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2019/124307 WO2021114089A1 (fr) 2019-12-10 2019-12-10 Procédés d'utilisation de crocétine dans le traitement de tumeurs solides

Country Status (2)

Country Link
CN (1) CN113993515B (fr)
WO (1) WO2021114089A1 (fr)

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004078695A1 (fr) * 2003-03-07 2004-09-16 Riken Vitamin Co., Ltd. Procede de purification de la crocetine
CN101157645A (zh) * 2007-11-27 2008-04-09 江南大学 一种藏红花酸的合成方法
US20100210572A1 (en) * 2009-02-18 2010-08-19 Thomas Eidenberger Hydrolysate of crocin
CN101863754A (zh) * 2009-04-17 2010-10-20 杭州法和医药科技有限公司 西红花酸、栀子黄色素和西红花色素的制备方法
JP2011098902A (ja) * 2009-11-05 2011-05-19 Riken Vitamin Co Ltd ジペプチジルペプチダーゼｉｖ阻害剤
WO2012060854A1 (fr) * 2010-11-02 2012-05-10 U.S Department Of Veterans Affairs Composé à base de crocétine purifiée et procédé de traitement, d'inhibition et/ou de prophylaxie du cancer et, notamment, du cancer du pancréas
US20140141082A1 (en) * 2012-11-16 2014-05-22 Song Gao Compositions Containing Enriched Natural Crocin and/or Crocetin, and Their Therapeutic or Nutraceutical Uses
CN109091458A (zh) * 2018-09-05 2018-12-28 嘉文丽（福建）化妆品有限公司 一种西红花酸微乳液及其冻干粉的制备方法

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104623670A (zh) * 2013-11-06 2015-05-20 高松 一种含西红花素的组合物及其应用

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004078695A1 (fr) * 2003-03-07 2004-09-16 Riken Vitamin Co., Ltd. Procede de purification de la crocetine
CN101157645A (zh) * 2007-11-27 2008-04-09 江南大学 一种藏红花酸的合成方法
US20100210572A1 (en) * 2009-02-18 2010-08-19 Thomas Eidenberger Hydrolysate of crocin
CN101863754A (zh) * 2009-04-17 2010-10-20 杭州法和医药科技有限公司 西红花酸、栀子黄色素和西红花色素的制备方法
JP2011098902A (ja) * 2009-11-05 2011-05-19 Riken Vitamin Co Ltd ジペプチジルペプチダーゼｉｖ阻害剤
WO2012060854A1 (fr) * 2010-11-02 2012-05-10 U.S Department Of Veterans Affairs Composé à base de crocétine purifiée et procédé de traitement, d'inhibition et/ou de prophylaxie du cancer et, notamment, du cancer du pancréas
US20140141082A1 (en) * 2012-11-16 2014-05-22 Song Gao Compositions Containing Enriched Natural Crocin and/or Crocetin, and Their Therapeutic or Nutraceutical Uses
CN109091458A (zh) * 2018-09-05 2018-12-28 嘉文丽（福建）化妆品有限公司 一种西红花酸微乳液及其冻干粉的制备方法

Also Published As

Publication number Publication date
CN113993515A (zh) 2022-01-28
CN113993515B (zh) 2023-08-29

Similar Documents

Publication Publication Date Title
RU2757373C2 (ru) Комбинированная терапия противоопухолевым алкалоидом
JP6404242B2 (ja) がんの治療のための併用療法としてのエリブリンおよびレンバチニブの使用
RU2738934C2 (ru) Способы лечения рака с использованием апилимода
EP3443962B1 (fr) Dérivé de quinoline pour le traitement du cancer de l'estomac
JP2012500180A5 (fr)
RU2731908C2 (ru) Композиции апилимода и способы его использования при лечениии меланомы
TW201713327A (zh) 治療癌症之方法
WO2017158570A1 (fr) Méthode de traitement d'un cancer du sein triple négatif
KR20230113594A (ko) Pik3ca 돌연변이 암을 치료하기 위한 병용 요법
EP3429582B1 (fr) Polythérapie pour traiter les maladies prolifératives
TW201713328A (zh) 治療癌症之方法
JP6462147B2 (ja) Ｈｓｐ９０阻害ペプチド結合体及びその腫瘍治療における応用
WO2020062951A1 (fr) Composé et utilisation associée
WO2021114089A1 (fr) Procédés d'utilisation de crocétine dans le traitement de tumeurs solides
JP7311177B2 (ja) Ａ－ＮＯＲ－５αアンドロスタン薬物と抗がん薬物との併用
TW201806592A (zh) 治療癌症的方法
JP7305197B2 (ja) 腫瘍転移を阻害する方法
CN109789136B (zh) 用于预防或治疗结肠直肠癌的药物联合组合物
BR112020009596A2 (pt) combinações de moduladores duplos de irs/stat3 e anticorpos anti pd-1/pd-l1 para tratar câncer
EP2211858B1 (fr) Utilisation de sels de pyridinium quaternaire pour inhiber des métastases cancéreuses
WO2024102968A1 (fr) Utilisations de composés antagonistes ep2/ep4 pour le traitement de polypose adénomateuse familiale (fap)
JP2023511072A (ja) 癌の治療のための組み合わせおよびその用途
US20090118340A1 (en) Method for inhibiting and/or preventing formation of cancer metastases and seconddary malignant cancers

Legal Events

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

Ref document number: 19955975

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 19955975

Country of ref document: EP

Kind code of ref document: A1