Pharmaceutical combination of an artemisinin compound, 5-aminolevulinic acid or methyl-5-aminolevulinic acid, and a chemotherapeutic agent

Abstract

The present invention relates to a pharmaceutical composition comprising an artemisinin compound, 5-aminolevulinic acid or methyl-5-aminolevulinic acid, and at least one chemotherapeutic agent, preferably at least one anti-glioblastoma drug. The pharmaceutical composition is used for the prevention and / or treatment of hematopoietic cancer, brain cancer, pancreatic cancer, liver cancer, breast cancer, and lung cancer, such as non-small cell lung cancer. Preferably, the present application provides a pharmaceutical composition comprising artemisinin (1a) or dihydroartemisinin (1b) or artesunate (1e), 5-aminolevulinic acid (2) or methyl-5-aminolevulinic acid (2b), and at least one chemotherapeutic agent, preferably at least one anti-glioblastoma drug, for use in the prevention and / or treatment of brain cancer, particularly glioblastoma.

Description

Detailed Description of the Invention

[0001] [Field of the Invention] The present invention relates to a pharmaceutical composition comprising an artemisinin compound or a pharmaceutically acceptable salt, cocrystal, or solvate thereof, 5-aminolevulinic acid or methyl-5-aminolevulinic acid or a pharmaceutically acceptable salt or solvate thereof, and at least one chemotherapeutic agent, preferably an anti-glioblastoma drug. The pharmaceutical composition is used for the prevention and / or treatment of hematopoietic cancer, brain cancer, pancreatic cancer, liver cancer, breast cancer, and lung cancer, such as non-small cell lung cancer. Preferably, the present application provides a pharmaceutical composition comprising artemisinin or dihydroartemisinin or artesunate, 5-aminolevulinic acid or methyl-5-aminolevulinic acid, and at least one chemotherapeutic agent, preferably at least one anti-glioblastoma drug, for use in the prevention and / or treatment of brain cancer, particularly glioblastoma.

[0002] More preferably, the present invention relates to a pharmaceutical composition comprising artemisinin (1a), artesunate (1e), or dihydroartemisinin (1b), and 5-aminolevulinic acid (2), and at least one chemotherapeutic agent, preferably one anti-glioblastoma drug, for use in the prevention and / or treatment of brain cancer, particularly glioblastoma.

[0003] [ka]

[0004] [Background of the invention] Artemisinin is a natural medicinal compound that can be isolated from the plant Artemisia annua, or sweet wormwood. This substance has been used in traditional Chinese medicine for thousands of years to treat fever, colds, and other illnesses. In the 1970s, artemisinin was first described as an antimalarial drug by the Chinese scientist Tu Youyou. Artemisinin, along with its derivatives, is one of the most commonly used antimalarials worldwide. Artemisinin kills the malaria-causing single-celled parasite P. falciparum in all its life stages. (Aweeka, FT & German, PI, Clinical Pharmacology of Artemisinin-Based Combination Therapies. Clinical Pharmacokinetics 47, 91-102 (2008)) Recently, the potent anti-cancer properties of artemisinin have been recognized. The variety of different functions of artemisinin and its uses indicates a broad mechanism of action of the compound in eukaryotic cells.

[0005] Artemisinin is a biochemically natural endoperoxide. This endoperoxide property is strictly required for its antimalarial effect. Upon cleavage of the endoperoxide bridge, artemisinin becomes activated, causing the production of reactive oxygen species (ROS) and free radicals, followed by alkylation of sensitive proteins and macromolecules in cells. This alkylation reaction alters protein structure and function, damages DNA, induces further cellular stress, and ultimately leads to cell death. Artemisinin binds nonspecifically to a wide range of cellular proteins and simultaneously alters multiple pathways, including glycolysis, protein biosynthesis, mitochondrial processes, and antioxidant responses. {Zhang, C.-J. et al. Haem-activated promiscuous targeting of artemisinin in Plasmodium falciparum. Nature Communications 6, 1-11 (2015)} {Ismail, H. et al. Artemisinin activity-based probes identify multiple molecular targets within the asexual stage of the malaria parasite Plasmodium falciparum 3D7. Proceedings of the National Academy of Sciences of the United States of America 113, 2080-2085 (2016)} The exact regulator or activator of artemisinin is still under debate, but free iron or iron complexes such as in the heme of hemoglobin are considered the most likely candidates.The promiscuous binding properties of artemisinin, along with its numerous reported functions, have so far prevented the identification of its distinct cellular targets. {Tilley, L., Straimer, J., Gnadig, NF, Ralph, SA, & Fidock, DA. Artemisinin Action and Resistance in Plasmodium falciparum. Trends in Parasitology 32, 682-696 (2016)} As an increasing number of cases of artemisinin-resistant malaria emerge, it is essential to clarify the mechanism of action of this compound to expand its applicability as a therapeutic.

[0006] 5-aminolevulinic acid (5-ALA) is an intermediate in heme biosynthesis and a tumor marker used to mark tumors for surgery. 5-aminolevulinic acid (5-ALA) is a non-protein amino acid.

[0007] The specific cytotoxicity of artemisinin against colorectal cancer (CRC) cells has been reported. Jigang Wang et al. showed that artemisinin / aminolevulinic acid combination therapy proved more effective than artemisinin monotherapy in a xenograft CRC model (ACS Cent. Sci. 2017, 3, pp. 743-750).

[0008] Glioblastoma, also known as glioblastoma multiforme (GBM), is the most aggressive cancer that develops in the brain. Glioblastoma accounts for 15% of brain tumors. Glioblastoma can begin in normal brain cells or develop from a pre-existing low-grade astrocytoma. Treatment typically involves surgery, followed by chemotherapy and radiation therapy. Glioblastoma is the most common cancer that develops in the brain and is the second most common brain tumor after meningioma. Approximately 3 people per 100,000 develop the disease annually. Glioblastoma most commonly develops around the age of 64 and is more common in men than women.

[0009] Despite maximal treatment, glioblastoma usually recurs. The typical survival time after diagnosis is 12-15 months, with fewer than 3-7% surviving longer than 5 years. Without treatment, survival is typically 3 months. There remains a great need for effective treatments for glioblastoma.

[0010] Therefore, it is an object of the present invention to provide a pharmaceutical composition useful for the prevention and / or treatment of cancer, particularly glioblastoma. The object of the present invention is solved by the teaching of the independent claims. Further advantageous features, aspects and details of the invention are evident from the dependent claims, the description, the drawings and the examples of the present application.

[0011] Surprisingly, it has been found in the present invention that a pharmaceutical composition comprising an artemisinin compound, and 5-aminolevulinic acid or methyl-5-aminolevulinic acid, and at least one chemotherapeutic agent, preferably one anti-glioblastoma drug, is useful for the prevention and / or treatment of cancer selected from hematopoietic cancer, brain cancer, pancreatic cancer, liver cancer, breast cancer, and lung cancer, such as non-small cell lung cancer, in particular glioblastoma.

[0012] The pharmaceutical composition may further comprise a pharmaceutically acceptable carrier, excipient and / or diluent. Optionally, the pharmaceutical composition is used in combination with radiation therapy, immunotherapy, electromagnetic field therapy, and / or hyperthermia.

[0013] [Summary of the Invention] The present invention provides a) artemisinin compound (1), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; b) 5-aminolevulinic acid (2), methyl-5-aminolevulinic acid (2b), or a pharmaceutically acceptable salt or solvate thereof; and c) at least one chemotherapeutic agent, preferably one anti-glioblastoma drug; The present invention provides a pharmaceutical composition comprising:

[0014] The artemisinin compound (1) is

[0015] [ka] or a pharmaceutically acceptable salt, co-crystal, or solvate of the artemisinin compound (1).

[0016] The chemotherapeutic agent, preferably an anti-glioblastoma drug, is preferably selected from the group comprising or consisting of temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevin, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, sertraline, irinotecan, clofazimine, and docetaxel.

[0017] The pharmaceutical compositions disclosed herein are useful as medicines, particularly for the prevention and / or treatment of hematopoietic cancers, brain cancer, pancreatic cancer, liver cancer, breast cancer, and lung cancer such as non-small cell lung cancer, preferably brain cancer.

[0018] Preferably, the glioblastoma is selected from proneural (PN), mesenchymal (MES), and classical (CL) glioblastoma subtypes.

[0019] For use in the prevention and / or treatment of hematopoietic cancer, brain cancer, pancreatic cancer, liver cancer, breast cancer, and lung cancer such as non-small cell lung cancer, preferably brain cancer, more preferably glioblastoma, in the pharmaceutical composition, the molar ratio of artemisinin compound to 5-aminolevulinic acid or methyl-5-aminolevulinic acid is preferably in the range of 1:5 to 1:5000, more preferably 1:5 to 1:1000, and even more preferably 1:10 to 1:500.

[0020] Optionally, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier, excipient, and / or diluent for use in the prevention and / or treatment of hematopoietic cancer, brain cancer, pancreatic cancer, liver cancer, breast cancer, and lung cancer such as non-small cell lung cancer, preferably brain cancer, more preferably glioblastoma.

[0021] In some embodiments, the pharmaceutical composition is used for the prevention and / or treatment of hematopoietic cancer, brain cancer, pancreatic cancer, liver cancer, breast cancer, and lung cancer such as non-small cell lung cancer, preferably brain cancer, more preferably glioblastoma, preferably in combination with radiation therapy, immunotherapy, electromagnetic field therapy, hyperthermia, chemotherapy, cancer immunotherapy, and / or any other small molecule-based therapy.

[0022] In some embodiments, the pharmaceutical composition is used in the form of a tablet, capsule, syrup, solution, suspension, emulsion, or gel.

[0023] In some embodiments, for the prevention and / or treatment of hematopoietic cancer, brain cancer, pancreatic cancer, liver cancer, breast cancer, and lung cancer such as non-small cell lung cancer, preferably brain cancer, more preferably glioblastoma, the pharmaceutical composition is administered by oral or parenteral application. Preferably, parenteral application includes intradermal application, intragastric application, intracutaneous application, intravascular application, intravenous application, intramuscular application, subcutaneous application, sublingual application, topical application, and transdermal application.

[0024] In some embodiments, for the prevention and / or treatment of hematopoietic cancer, brain cancer, pancreatic cancer, liver cancer, and lung cancer such as non-small cell lung cancer, preferably brain cancer, more preferably glioblastoma, the artemisinin contained in the pharmaceutical composition is administered in the range of 0.01 to 100 mg / kg of body weight per day, and the 5-aminolevulinic acid is administered in the range of 0.01 to 200 mg / kg of body weight per day.

[0025] In some embodiments, the pharmaceutical composition is preferably used for the prevention and / or treatment of glioblastoma in combination with at least one chemotherapeutic agent, preferably one anti-glioblastoma drug, wherein the at least one chemotherapeutic agent, preferably the at least one anti-glioblastoma drug, is administered in the range of 0.01 to 100 mg / kg of body weight per day.

[0026] [Description of the Invention] The present invention provides a) artemisinin compound (1), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; and b) 5-aminolevulinic acid (2), methyl-5-aminolevulinic acid (2b), or a pharmaceutically acceptable salt or solvate thereof; and c) at least one chemotherapeutic agent, preferably one anti-glioblastoma drug; The present invention provides a pharmaceutical composition comprising:

[0027] Surprisingly, the pharmaceutical compositions disclosed herein, which comprise three pharmaceutically active ingredients, namely, an artemisinin compound (1), and 5-aminolevulinic acid (2) or methyl-5-aminolevulinic acid (2b), and at least one chemotherapeutic agent, preferably at least one anti-glioblastoma drug, are significantly more effective than the combination of only two of these active ingredients, as is evident from Figures 13-19.

[0028] The term "artemisinin compound" as used herein refers to artemisinin (1a), dihydroartemisinin (1b), artemether (1c), arteether (1d), and artesunate (1e), as well as pharmaceutically acceptable salts, co-crystals, or solvates of artemisinin compound (1), as shown below.

[0029] [ka]

[0030] In all pharmaceutical compositions disclosed herein, artemisinin (1a), dihydroartemisinin (1b), and artesunate (1e) are preferred as the artemisinin compound (1). More preferred are artemisinin (1a) and artesunate (1e), and most preferred is artemisinin (1a).

[0031] Therefore, the present application preferably provides: a) an artemisinin compound (1) selected from the group consisting of artemisinin (1a), dihydroartemisinin (1b), and artesunate (1e), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; b) 5-aminolevulinic acid (2), methyl-5-aminolevulinic acid (2b), or a pharmaceutically acceptable salt or solvate thereof; and c) at least one chemotherapeutic agent, preferably one anti-glioblastoma drug; The present invention relates to a pharmaceutical composition comprising:

[0032] More preferably, the present application provides a) artemisinin (1a) or artesunate (1e), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; b) 5-aminolevulinic acid (2), methyl-5-aminolevulinic acid (2b), or a pharmaceutically acceptable salt or solvate thereof; and c) at least one chemotherapeutic agent, preferably one anti-glioblastoma drug; The present invention relates to a pharmaceutical composition comprising:

[0033] Even more preferably, the present application provides a) artesunate (1e), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; b) 5-aminolevulinic acid (2), methyl-5-aminolevulinic acid (2b), or a pharmaceutically acceptable salt or solvate thereof; and c) at least one chemotherapeutic agent, preferably one anti-glioblastoma drug; The present invention relates to a pharmaceutical composition comprising:

[0034] Even more preferably, the present application provides a) artemisinin (1a), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; b) 5-aminolevulinic acid (2), methyl-5-aminolevulinic acid (2b), or a pharmaceutically acceptable salt or solvate thereof; and c) at least one chemotherapeutic agent, preferably one anti-glioblastoma drug; The present invention relates to a pharmaceutical composition comprising:

[0035] With regard to component b), 5-aminolevulinic acid (2) is preferred over methyl-5-aminolevulinic acid (2b).

[0036] [ka]

[0037] Therefore, the present invention provides a) artemisinin compound (1), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; and b) 5-aminolevulinic acid (2), or a pharmaceutically acceptable salt or solvate thereof; and c) at least one chemotherapeutic agent, preferably one anti-glioblastoma drug; The present invention provides a pharmaceutical composition comprising:

[0038] Furthermore, the present application preferably provides a) an artemisinin compound (1) selected from the group consisting of artemisinin (1a), dihydroartemisinin (1b), and artesunate (1e), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; and b) 5-aminolevulinic acid (2), or a pharmaceutically acceptable salt or solvate thereof; and c) at least one chemotherapeutic agent, preferably one anti-glioblastoma drug; The present invention relates to a pharmaceutical composition comprising:

[0039] More preferably, the present application provides a) artemisinin (1a) or artesunate (1e), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; and b) 5-aminolevulinic acid (2), or a pharmaceutically acceptable salt or solvate thereof; and c) at least one chemotherapeutic agent, preferably one anti-glioblastoma drug; The present invention relates to a pharmaceutical composition comprising:

[0040] Even more preferably, the present application provides a) artesunate (1e), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; and b) 5-aminolevulinic acid (2), or a pharmaceutically acceptable salt or solvate thereof; and c) at least one chemotherapeutic agent, preferably one anti-glioblastoma drug; The present invention relates to a pharmaceutical composition comprising:

[0041] Even more preferably, the present application provides a) artemisinin (1a), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; and b) 5-aminolevulinic acid (2), or a pharmaceutically acceptable salt or solvate thereof; and c) at least one chemotherapeutic agent, preferably one anti-glioblastoma drug; The present invention relates to a pharmaceutical composition comprising:

[0042] The chemotherapeutic agent, preferably an anti-glioblastoma drug, is preferably selected from the group comprising or consisting of temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevin, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, sertraline, irinotecan, clofazimine, and docetaxel.

[0043] More preferably, the anti-glioblastoma drug is selected from the group including or consisting of temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevine, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, and sertraline.

[0044] Even more preferably, the anti-glioblastoma drug is selected from the group including or consisting of temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, and vinorevin.

[0045] Even more preferably, the anti-glioblastoma drug is selected from the group including or consisting of temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, flubendazole, vindesine sulfate, cerivastatin, and vinorevin.

[0046] Even more preferably, the anti-glioblastoma drug is selected from the group including or consisting of temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, ribavirin, and vinorebin.

[0047] Even more preferably, the anti-glioblastoma drug is selected from the group including or consisting of temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, and vinorebin.

[0048] Even more preferably, the anti-glioblastoma drug is selected from the group including or consisting of temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, and doxorubicin.

[0049] Even more preferably, the anti-glioblastoma agent is selected from the group comprising or consisting of temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, and dactinomycin.

[0050] Even more preferably, the anti-glioblastoma agent is selected from the group comprising or consisting of temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, and homoharringtonine.

[0051] Even more preferably, the anti-glioblastoma agent is selected from the group comprising or consisting of temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and homoharringtonine.

[0052] Even more preferably, the anti-glioblastoma agent is selected from the group comprising or consisting of temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and triptolide.

[0053] Even more preferably, the anti-glioblastoma drug is selected from the group comprising or consisting of temozolomide, lomustine, cisplatin, and 5-fluorouracil. Most preferably, it is temozolomide.

[0054] Preliminary data indicate that the combination of artemisinin, 5-aminolevulinic acid, and triptolide or homoharringtonine or dactinomycin or doxorubicin or epirubicin or vinorebin is comparable to the combination of artemisinin, 5-aminolevulinic acid, and temozolomide.

[0055] Accordingly, the present invention provides a pharmaceutical composition comprising the following a to c: a) artemisinin compound (1), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; and b) 5-aminolevulinic acid (2), methyl-5-aminolevulinic acid (2b), or a pharmaceutically acceptable salt or solvate thereof; and c) temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevin, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, sertraline, irinotecan, clofazimine, and docetaxel; Preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevine, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, and sertraline; More preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, flubendazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, ribavirin, and vinorebin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, and doxorubicin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, and dactinomycin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and triptolide; Even more preferably, at least one anti-glioblastoma agent selected from the group comprising or consisting of temozolomide, lomustine, cisplatin, and 5-fluorouracil, most preferably temozolomide.

[0056] Furthermore, the present application preferably relates to a pharmaceutical composition comprising the following a to c: a) an artemisinin compound (1) selected from the group consisting of artemisinin (1a), dihydroartemisinin (1b), and artesunate (1e), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; b) 5-aminolevulinic acid (2), methyl-5-aminolevulinic acid (2b), or a pharmaceutically acceptable salt or solvate thereof; and c) temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevin, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, sertraline, irinotecan, clofazimine, and docetaxel; Preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevine, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, and sertraline; More preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, flubendazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, ribavirin, and vinorebin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, and doxorubicin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, and dactinomycin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and triptolide; Even more preferably, at least one anti-glioblastoma agent selected from the group comprising or consisting of temozolomide, lomustine, cisplatin, and 5-fluorouracil, most preferably temozolomide.

[0057] More preferably, the present application relates to a pharmaceutical composition comprising the following a to c: a) artemisinin (1a), or artesunate (1e), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; b) 5-aminolevulinic acid (2), methyl-5-aminolevulinic acid (2b), or a pharmaceutically acceptable salt or solvate thereof; and c) temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevin, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, sertraline, irinotecan, clofazimine, and docetaxel; Preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevine, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, and sertraline; More preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, flubendazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, ribavirin, and vinorebin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, and doxorubicin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, and dactinomycin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and triptolide; Even more preferably, at least one anti-glioblastoma agent selected from the group comprising or consisting of temozolomide, lomustine, cisplatin, and 5-fluorouracil, most preferably temozolomide.

[0058] Even more preferably, the present application relates to a pharmaceutical composition comprising: a) artesunate (1e), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; b) 5-aminolevulinic acid (2), methyl-5-aminolevulinic acid (2b), or a pharmaceutically acceptable salt or solvate thereof; and c) temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevin, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, sertraline, irinotecan, clofazimine, and docetaxel; Preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevine, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, and sertraline; More preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, flubendazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, ribavirin, and vinorebin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, and doxorubicin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, and dactinomycin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and triptolide; Even more preferably, at least one anti-glioblastoma agent selected from the group comprising or consisting of temozolomide, lomustine, cisplatin, and 5-fluorouracil, most preferably temozolomide.

[0059] Even more preferably, the present application relates to a pharmaceutical composition comprising: a) artemisinin (1a), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; b) 5-aminolevulinic acid (2), methyl-5-aminolevulinic acid (2b), or a pharmaceutically acceptable salt or solvate thereof; and c) temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevin, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, sertraline, irinotecan, clofazimine, and docetaxel; Preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevine, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, and sertraline; More preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, flubendazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, ribavirin, and vinorebin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, and doxorubicin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, and dactinomycin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and triptolide; Even more preferably, at least one anti-glioblastoma agent selected from the group comprising or consisting of temozolomide, lomustine, cisplatin, and 5-fluorouracil, most preferably temozolomide.

[0060] Preferably, the present invention provides a pharmaceutical composition comprising the following a to c: a) artemisinin compound (1), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; and b) 5-aminolevulinic acid (2), or a pharmaceutically acceptable salt or solvate thereof; and c) temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevin, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, sertraline, irinotecan, clofazimine, and docetaxel; Preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevine, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, and sertraline; More preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, flubendazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, ribavirin, and vinorebin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, and doxorubicin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, and dactinomycin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and triptolide; Even more preferably, at least one anti-glioblastoma agent selected from the group comprising or consisting of temozolomide, lomustine, cisplatin, and 5-fluorouracil, most preferably temozolomide.

[0061] Furthermore, the present application preferably relates to a pharmaceutical composition comprising the following a to c: a) an artemisinin compound (1) selected from the group consisting of artemisinin (1a), dihydroartemisinin (1b), and artesunate (1e), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; b) 5-aminolevulinic acid (2), or a pharmaceutically acceptable salt or solvate thereof; and c) temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevin, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, sertraline, irinotecan, clofazimine, and docetaxel; Preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevine, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, and sertraline; More preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, flubendazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, ribavirin, and vinorebin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, and doxorubicin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, and dactinomycin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and triptolide; Even more preferably, at least one anti-glioblastoma agent selected from the group comprising or consisting of temozolomide, lomustine, cisplatin, and 5-fluorouracil, most preferably temozolomide.

[0062] More preferably, the present application relates to a pharmaceutical composition comprising the following a to c: a) artemisinin (1a), or artesunate (1e), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; b) 5-aminolevulinic acid (2), or a pharmaceutically acceptable salt or solvate thereof; and c) temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevin, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, sertraline, irinotecan, clofazimine, and docetaxel; Preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevine, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, and sertraline; More preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, flubendazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, ribavirin, and vinorebin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, and doxorubicin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, and dactinomycin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and triptolide; Even more preferably, at least one anti-glioblastoma agent selected from the group comprising or consisting of temozolomide, lomustine, cisplatin, and 5-fluorouracil, most preferably temozolomide.

[0063] Even more preferably, the present application relates to a pharmaceutical composition comprising: a) artesunate (1e), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; b) 5-aminolevulinic acid (2), or a pharmaceutically acceptable salt or solvate thereof; and c) temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevin, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, sertraline, irinotecan, clofazimine, and docetaxel; Preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevine, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, and sertraline; More preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, flubendazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, ribavirin, and vinorebin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, and doxorubicin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, and dactinomycin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and triptolide; Even more preferably, at least one anti-glioblastoma agent selected from the group comprising or consisting of temozolomide, lomustine, cisplatin, and 5-fluorouracil, most preferably temozolomide.

[0064] Even more preferably, the present application relates to a pharmaceutical composition comprising: a) artemisinin (1a), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; and b) 5-aminolevulinic acid (2), or a pharmaceutically acceptable salt or solvate thereof; and c) temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevin, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, sertraline, irinotecan, clofazimine, and docetaxel; Preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevine, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, and sertraline; More preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, flubendazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, ribavirin, and vinorebin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, and doxorubicin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, and dactinomycin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and triptolide; Even more preferably, at least one anti-glioblastoma agent selected from the group comprising or consisting of temozolomide, lomustine, cisplatin, and 5-fluorouracil, most preferably temozolomide.

[0065] As used herein, the term "artemisinin compound" refers to a compound selected from the group including or consisting of artemisinin (ART or Art or 1a), artemether, arteether, artesunate (ARS or 1e), and dihydroartemisinin (arteminol or DHA or 1b).

[0066] [ka]

[0067] The active metabolite of artemisinin compound (1) is generally dihydroartemisinin (DHA). As used herein, the terms "artemisinin," "ART," and "Art" are used interchangeably and include the well-known compound having the chemical structure (1a).

[0068] As used herein, and unless otherwise specified, the term "pharmaceutically acceptable salts of artemisinin compound (1)" includes, but is not limited to, salts of the acidic or basic moieties of the compounds described herein. The basic moieties can form a variety of salts with various inorganic and organic acids. Acids that can be used to prepare pharmaceutically acceptable acid addition salts of the basic compounds are those that form non-toxic acid addition salts, for example, salts with pharmacologically acceptable anions. Suitable organic acids include, but are not limited to, maleic acid, fumaric acid, benzoic acid, ascorbic acid, succinic acid, acetic acid, formic acid, oxalic acid, propionic acid, tartaric acid, salicylic acid, citric acid, gluconic acid, lactic acid, mandelic acid, cinnamic acid, oleic acid, tannic acid, aspartic acid, stearic acid, palmitic acid, glycolic acid, glutamic acid, gluconic acid, glucaronic acid, saccharic acid, isonicotinic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, or pamoic acid (e.g., 1,1'-methylene-bis-(2-hydroxy-3-naphthoate)) acid. Suitable inorganic acids include, but are not limited to, hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid, or nitric acid. Compounds containing an amine moiety can form pharmaceutically acceptable salts with various amino acids in addition to the acids mentioned above. Chemical moieties that are acidic in nature are capable of forming base salts with various pharmacologically acceptable cations, such as alkali metal or alkaline earth metal salts, particularly calcium, magnesium, sodium, lithium, zinc, potassium, or iron salts.

[0069] As used herein, and unless otherwise specified, the term "pharmaceutically acceptable salts of 5-aminolevulinic acid (2)" or "pharmaceutically acceptable salts of methyl-5-aminolevulinic acid (2b)" includes, but is not limited to, salts of the acidic or basic moieties of the compounds described herein. The basic moieties are capable of forming a variety of salts with various inorganic and organic acids. Acids that can be used to prepare pharmaceutically acceptable acid addition salts of the basic compounds are those that form non-toxic acid addition salts, for example, salts with pharmacologically acceptable anions. Suitable organic acids include, but are not limited to, maleic acid, fumaric acid, benzoic acid, ascorbic acid, succinic acid, acetic acid, formic acid, oxalic acid, propionic acid, tartaric acid, salicylic acid, citric acid, gluconic acid, lactic acid, mandelic acid, cinnamic acid, oleic acid, tannic acid, aspartic acid, stearic acid, palmitic acid, glycolic acid, glutamic acid, gluconic acid, glucaronic acid, saccharic acid, isonicotinic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, or pamoic acid (e.g., 1,1'-methylene-bis-(2-hydroxy-3-naphthoate)) acid. Suitable inorganic acids include, but are not limited to, hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid, or nitric acid. Compounds containing an amine moiety can form pharmaceutically acceptable salts with various amino acids in addition to the acids mentioned above. Chemical moieties that are acidic in nature are capable of forming base salts with various pharmacologically acceptable cations, such as alkali metal or alkaline earth metal salts, particularly calcium, magnesium, sodium, lithium, zinc, potassium, or iron salts.

[0070] As used herein, and unless otherwise specified, the term "cocrystal," "pharmaceutically acceptable cocrystal of an artemisinin compound," refers to a crystalline material composed of two or more different molecules, i.e., an artemisinin compound as the active pharmaceutical ingredient (API) and a cocrystal former ("coformer"), within the same crystal lattice.

[0071] Because cocrystals are primarily hydrogen bond formation between drug molecules and coformers, any API can potentially cocrystallize regardless of whether it has acidic, basic, or ionizable groups. Cocrystallization can improve physicochemical properties such as solubility, dissolution rate, chemical stability, and melting point. The interactions involved in cocrystal formation include hydrogen bonding, π-stacking, and intermolecular forces.

[0072] Preferred coformers for cocrystals of artemisinin compounds may include, but are not limited to, nicotinamide, ascorbic acid, urea, tromethamine, theophylline, theophylline-7-acetate, theobromine, sulfamide, sucrose, sorbitol, saccharin, pyridoxine, phloroglucinol, paracetamol, N-methylglucosamine, methanesulfonic acid, D-mannitol, and malonic acid.

[0073] As used herein, and unless otherwise specified, the term "solvate" refers to a compound that further includes a stoichiometric or non-stoichiometric amount of solvent bound by non-covalent intermolecular forces. The solvent may include, but is not limited to, alcohols such as ethanol, propanol, isopropanol, n-butanol, glycol, N-methyl-2-pyrrolidone, acetonitrile, N,N'-dimethylformamide, dimethyl sulfoxide, water, etc. When the solvent is water, the solvate is a hydrate.

[0074] As used herein, the terms "5-aminolevulinic acid," "delta-aminolevulinic acid," "ALA," and "5-ALA" are used interchangeably and include 5-amino-4-oxopentanoic acid of chemical structure (2). When used in reference to ALA, the term "derivative" includes chemically modified compounds, such as esters such as ALA esters. Such compounds are typically formed by modifying the carboxylic acid group of 5-ALA. The term "derivative" is also intended to include any 5-aminolevulinic acid compound, where the 5-amino group may be substituted or unsubstituted. In other words, the term "derivative" used in reference to 5-ALA includes compounds in which chemical modification occurs at either the carboxylic acid or the 5-amino group of ALA, or both. Derivatives of 5-ALA are generally known and are described in the prior art, for example, in WO96 / 28412, WO02 / 10120, and WO2005 / 092838. Preferred derivatives of 5-ALA for use in the present invention are ALA esters, particularly esters of 5-ALA compounds in which the 5-amino group is unsubstituted.

[0075] In humans, 5-aminolevulinic acid is a precursor of heme. 5-aminolevulinic acid undergoes a series of transformations in the cytosol and is ultimately converted to protoporphyrin IX in mitochondria. This protoporphyrin molecule chelates with iron in the presence of the enzyme ferrochelatase to produce heme. Therefore, administration of 5-aminolevulinic acid can be used to increase intracellular heme levels and intracellular iron concentrations. 5-aminolevulinic acid can be administered per se or in any pharmaceutically acceptable physical form. For example, 5-aminolevulinic acid can be in the form of a pharmaceutically acceptable salt or solvate.

[0076] As used herein, the term "pharmaceutical salt of 5-aminolevulinic acid" typically refers to a salt obtained from ALA or an ALA derivative and a monoprotic acid, such as a sulfonic acid, such as methanesulfonic acid, thereby forming a 1:1 salt. Alternatively, a salt may be formed between ALA or an ALA derivative and a diprotic or triprotic acid, such as a sulfonic acid, such as ethane-1,2-disulfonic acid, sulfuric acid, or phosphoric acid. When an acid having more than one acidic proton is used, the resulting compound may have a stoichiometric ratio other than 1:1, such as 2:1 (ALA:acid) or 3:1 (ALA:acid), or may include a mixture of salts with various levels of stoichiometry. Sulfuric acid may form, for example, a 2:1 (ALA:acid) salt, while phosphoric acid may form a 3:1 (ALA:acid) salt. Polybasic acids may also form other salts with ALA or an ALA derivative. For example, sulfuric acid may be converted to HSO4. - A 1:1 (ALA:acid) salt based on the anion may be provided, where phosphate is HPO4 2- and H2PO4 -Both 2:1 (ALA:acid) salts and 1:1 (ALA:acid) salts (or a combination thereof) based on the respective anions may be provided. Furthermore, polybasic acids can also form other salts, such as 1:1 salts, with ALA or ALA derivatives (e.g., ALA esters) in the form of salts with other physiologically acceptable bases, such as sodium hydroxide, calcium hydroxide, potassium hydroxide, and meglumine. The salts of the present invention are preferably derived from acids having a pKa of about 4 or less, more preferably about 3 or less. The acid may be inorganic or organic. Preferred inorganic acids include hydrobromic acid, hydrochloric acid, sulfuric acid, phosphoric acid, and nitric acid. Preferred organic acids include sulfonic acid and sulfonic acid derivatives. Particularly preferred are salts derived from hydrochloric acid, nitric acid, sulfonic acid, and sulfonic acid derivatives. The term "sulfonic acid" is intended to include any organic compound containing at least one -SO3H group. Preferably, it may contain 1, 2 or 3 -SO3H groups, most preferably 1 or 2, e.g., 1. When used in reference to sulfonic acids, the term "derivative" is intended to encompass any such compound containing at least one (preferably 1, 2 or 3, most preferably 1 or 2, e.g., 1) -SO3X group (wherein X is a physiologically acceptable cation, such as, for example, sodium, calcium, potassium, magnesium or meglumine cation).

[0077] As used herein, the term "treating" or "treatment" includes reversing, alleviating, or inhibiting the progression of the disease, disorder, or condition to which the term applies, or ameliorating one or more symptoms of the disease, disorder, or condition. As used herein, "treating" or "treatment" may also refer to reducing the likelihood or incidence of a disease, disorder, or condition in a mammal compared to an untreated control population or compared to the same mammal prior to treatment. For example, as used herein, "treating" may refer to preventing a disease, disorder, or condition, and may include delaying or preventing the onset of a disease, disorder, or condition, or may include delaying or preventing symptoms associated with a disease, disorder, or condition. As used herein, "treating" may also refer to reducing the severity of a disease, disorder, or condition associated with the disease, disorder, or condition prior to onset in a mammal having the disease, disorder, or condition. Such prevention or reduction of the severity of a disease, disorder, or condition prior to onset relates to administering a composition of the invention described herein to a subject who is not suffering from the disease, disorder, or condition at the time of administration. As used herein, the term "treating" may also refer to preventing the recurrence of the disease, disorder, or condition, or the recurrence of one or more symptoms associated with the disease, disorder, or condition. As used herein, the terms "treatment" and "therapeutically" refer to the act of treating, as "treating" is defined above.

[0078] Preferably, the disease or disorder is cancer selected from hematopoietic cancer, brain cancer, pancreatic cancer, liver cancer, breast cancer, and lung cancer such as non-small cell lung cancer, preferably brain cancer, in particular glioblastoma.

[0079] The term "effective amount" as used herein refers to the amount required for an untreated patient to treat a cancer selected from hematopoietic cancer, brain cancer, pancreatic cancer, liver cancer, breast cancer, and lung cancer such as non-small cell lung cancer, preferably brain cancer, particularly glioblastoma. The effective amount of the active compound used to practice the present invention for the therapeutic treatment of a disease varies depending on the method of administration, the age, weight, and general health of the subject. Ultimately, the attending physician or veterinarian will determine the appropriate amount and administration schedule. This amount is referred to as an "effective" amount.

[0080] The term "patient" or "subject" as used herein refers to a mammalian subject (primate (e.g., human, cow, sheep, goat, pig, horse, dog, cat, rabbit, rat, mouse, etc.), preferably a human subject, who has, is suspected of having, or is susceptible to, or may have a condition associated with cancer selected from hematopoietic cancer, brain cancer, pancreatic cancer, liver cancer, breast cancer, and lung cancer such as non-small cell lung cancer, preferably brain cancer, particularly glioblastoma. In one embodiment, the method may be used to treat a patient suffering from multiple sclerosis, for example, a patient having any of the conditions described above. In other embodiments, the method may also be used to prevent a perspective patient from developing glioblastoma.

[0081] Glioblastoma, also known as glioblastoma multiforme (GBM), is the most aggressive cancer that occurs in the brain.Glioblastoma has the subtypes of proneural (PN), mesenchymal (MES) and classical (CL) glioblastoma.Therefore, in one embodiment, glioblastoma is preferably selected from the subtypes of proneural (PN), mesenchymal (MES) and classical (CL) glioblastoma.

[0082] We conducted a screen to identify artemisinin targets in haploid mouse stem cells (haESCs) using genome-wide mutagenesis (Figure 1A). To establish screening conditions, we determined the LD values of compounds in wild-type haESCs and evaluated their proliferation rates in different mouse tumor cell lines (Figures 5A and 5B). To generate genome-wide mutant ESC libraries, we used two targeting systems for insertional mutagenesis: retrovirus and Tol2 transposon (Schnutgen, F. et al., Enhanced gene trapping in mouse embryonic stem cells. Nucleic Acids Research 36, e133-e133 (2008)). Selection of these independent libraries resulted in the recovery of artemisinin-resistant colonies from the mutagenized cell pool but not from the control cells. Upon expansion and mapping of their insertion sites, we determined enrichment scores based on loss-of-function (LOF) analysis.

[0083] Haploid mutagenesis and artemisinin compound screening identified the enzyme protoporphyrinogen oxidase (PPOX) as the top hit in both mutant libraries. PPOX is an inner mitochondrial membrane enzyme involved in cellular porphyrin or heme production. Interestingly, genetic variants of every single enzyme in the porphyrin biosynthetic pathway, both mitochondrial (Alas1, Cpox, Ppox, Fech) and cytosolic (Alad, Hmbs, Uros, Urod) pathways, as well as essentially all relevant cofactor-generating enzymes and pathways that contribute to protoporphyrin or heme biosynthesis (Lias, Ogdh, Dlst, Lip1, Lipt2, Pdxk) were recovered from the screening (Figure 1B; Figure 5C). GO term analysis of the artemisinin screening profile in ESCs confirmed porphyrin biosynthesis as the most important target pathway in relation to artemisinin toxicity (Figure 1C). Thus, forward genetic screening in haploid ESCs illustrates porphyrin biosynthesis as a crucial and essential pathway for artemisinin cytotoxicity.

[0084] To confirm hits from the high-throughput artemisinin screen at the individual level, we tested single mutant ESC clones for sensitivity to artemisinin. Individual colonies were recovered from both mutant libraries directly from the screen and used to establish single cell lines. From these, we generated labeled wild-type (mCherry_Cre) and knockout (GFP) sister cell clones (via Cre-recombinase-mediated reversion of the knockout cassette) and monitored their relative growth rates in the presence of artemisinin. While the ratio of mixed labeled cells remained constant in the absence of compound or in control cells, artemisinin exposure resulted in the selective loss of wild-type (mCherry_Cre) and the robust expansion of knockout (GFP) cells. Insertion site mapping (using Sanger sequencing) in resistant cells confirmed the disruption of porphyrin biosynthetic pathway components (Figures 1-2D). These results confirm that mutations in enzymes of the porphyrin biosynthetic pathway induce artemisinin resistance in mouse ESCs.

[0085] Applicant next evaluated whether modulation of cellular porphyrin biosynthesis was sufficient to alter sensitivity to artemisinin. Applicant used the protoporphyrinogen oxidase (PPOX) inhibitor acifluorfen, a pharmacological inhibitor of porphyrin production or heme biosynthesis that inhibits the conversion of protoporphyrinogen IX to protoporphyrin IX, and observed increased resistance to artemisinin in mouse ESCs (Figure 2-1A). {Witkowski, DA & Halling, BP. Inhibition of plant protoporphyrinogen oxidase by the herbicide acifluorfen-methyl. Plant Physiol. 90, 1239-1242 (1989)}. Conversely, when Applicant enhanced porphyrin production using a non-proteinogenic amino acid and the endogenous alanine analog precursor δ-aminolevulinic acid (5-ALA), it increased susceptibility to anti-malarial compounds (Figure 2B). Applicant next analyzed whether different mouse and human cancer cell lines respond similarly to combination therapy, regardless of basal growth and susceptibility rates (Figure 5B).

[0086] The applicant not only confirmed that all tested cancer cells acquired hypersensitivity to artemisinin (Figure 2-1C; Figure 6-1A). Surprisingly, 5-ALA also induced artemisinin hypersensitivity to artemisinin cytotoxicity in three independently generated clinically relevant human primary glioblastoma cell lines (Figure 2-1D; Figure 6-1B, 6-1C).

[0087] Mechanistically, artemisinin has previously been shown to induce reactive oxygen species (ROS) production. {Gopalakrishnan, AM & Kumar, N. Antimalarial action of artesunate involves DNA damage mediated by reactive oxygen species. Antimicrob. Agents Chemother. 59, 317-325 (2015)} {Stockwin, L. Het al. Artemisinin dimer anticancer activity correlates with heme-catalyzed reactive oxygen species generation and endoplasmic reticulum stress induction. Int. J. Cancer 125, 1266-1275 (2009)} {Berman, PA & Adams, PA. Artemisinin enhances heme-catalyzed oxidation of lipid membranes. Free Radic. Biol. Med. 22, 1283-1288 (1997)}. Assessment of ROS levels (using the redox-sensitive fluorescent probe dihydroethidium (DHE)) confirmed an increase in ROS levels upon artemisinin treatment [Figures 6-1D and 6-2E]. Applicant next evaluated whether 5-ALA had any effect on ROS in the presence of artemisinin. Notably, 5-ALA alone caused ROS and a slight decrease in cell viability. However, the combination of 5-ALA and artemisinin resulted in highly elevated ROS levels and a strong increase in cell death in two different cellular contexts [Figures 2E and 2F; Figures 6F and 6G].Similarly, mitochondrial polarization (ΔΨm, as assessed by the mitochondrial membrane potential probe JC-1), previously associated with artemisinin-induced ROS induction, was strongly elevated in combination with 5-ALA [Figure 6-2H]. {Antoine, T. et al. Rapid kill of malaria parasites by artemisinin and semi-synthetic endoperoxides involves ROS-dependent depolarization of the membrane potential. J. Antimicrob. Chemother. 69, 1005-1016 (2014).} Notably, 5-ALA treatment alone increased mitochondrial polarization, indicating sensitization to concurrent artemisinin treatment. Importantly, all of the named phenotypes (ROS induction, mitochondrial depolarization, and cell death) could be suppressed by pharmacological inhibition of porphyrin production [Figures 2E and 2F; Figures 6F, 6G, and 6H]. These results demonstrated that artemisinin and 5-ALA can induce elevated ROS levels (and mitochondrial membrane depolarization) and reverse cell death by inhibiting porphyrin biosynthesis.

[0088] Alterations in cellular metabolism and upregulation of porphyrin production are frequently observed in human cancer cells. (Navone, NM, Polo, CF, Frisardi, AL, Andrade, NE & Battle, AM. Heme biosynthesis in human breast cancer—mimetic 'in vitro' studies and some heme enzymic activity levels. Int. J. Biochem. 22, 1407-1411 (1990).) In human glioblastoma patients, porphyrin precursors (i.e., protoporphyrins) exhibit strong fluorescence that can be monitored and functionally utilized, and thus elevated porphyrin biosynthesis can be used therapeutically to locate and target cancerous tissue. (Zhao, S. et al. Intraoperative fluorescence-guided resection of high-grade malignant gliomas using 5-aminolevulinic acid-induced porphyrins: a systematic review and meta-analysis of prospective studies, PloS One 8, e63682 (2013)) Fluorescent protoporphyrins, such as protoporphyrin IX, specifically accumulate in tumor cells, so oral or intravenous administration of 5-ALA during tumor tissue resection in the clinic can be used to distinguish malignant tumor tissue from healthy brain material. {Marbacher, S. et al. Use of fluorescence to guide resection or biopsy of primary brain tumors and brain metastases. Neurosurg Focus 36, E10 (2014).}Therefore, applicants evaluated whether the endogenously altered porphyrin biosynthesis in brain tumors could be therapeutically exploited using a combination of 5-ALA and artemisinin.

[0089] We use a recently established human brain tumor organoid model, which is based on genetic manipulation of neuronal precursors during cerebral organoid development and thus recapitulates key aspects of human tumorigenesis in vitro (Lancaster, MA et al. Cerebral organoids model human brain development and microcephaly. Nature 501, 373-379 (2013)). Tumor cells are simultaneously labeled with GFP, enabling spatial monitoring over time and in vitro compound profiling in human brain tumor organoids (Figure 3-1A). We first evaluated the growth rate in the presence of artemisinin and 5-ALA in a central nervous system primitive neuroectodermal tumor (CNS-PNET)-like neoplasia model based on overexpression of the c-MYC oncogene {Bian, S. et al. Genetically engineered cerebral organoids model brain tumor formation. Nat. Methods 15, 631-639 (2018)} [Figure 7-1A]. Highly malignant brain tumor organoids were treated with different doses of 5-ALA, artemisinin, or a combination of both [Figure 7-1B]. Because very high concentrations of 5-ALA or artemisinin alone resulted in significant growth inhibition of GFP-positive tumor organoids [Figure 7-1A], concentrations of individual compounds that did not significantly affect tumor or organoid growth were selected for all subsequent experiments. The combination of 5-ALA and artemisinin significantly reduced GFP-positive tumor cells in human brain organoids [Figures 3C and 3D; Figure 7B]. Quantification of tumor tissue area at different time points (d3 and d5) [Figure 3E; Figure 7C] and FACS analysis (d5) [Figure 3F] confirmed the reduction of GFP-positive cells in the combined treatment of 5-ALA and artemisinin.Furthermore, immunohistochemistry of brain organoid sections showed that 5-ALA treatment resulted in the loss of GFP+ tumor tissue but had no apparent effect on non-transformed neuronal tissue in brain organoids, such as Sox2-expressing cells and / or rosette-like structure indicator cells or progenitor cells (Figure 7-4D, Figures 7-1-7-4). In summary, this data suggests that 5-ALA and artemisinin combination therapy significantly reduces the number of tumor cells in a human in vitro primitive neuroectodermal tumor (PNET) organoid model.

[0090] Since we observed a significant effect of 5-ALA and artemisinin on ROS levels, we performed DHE staining and Myc-overexpressing tumor organoid analysis. While GFP-positive tumor cells showed slightly higher basal ROS signals compared with GFP-negative wild-type cells (assessed by DHE staining and FACS analysis), ROS levels were further significantly increased by 5-ALA and artemisinin dual treatment, especially in GFP-positive cancer cells (Figure 3G; Figure 9A). Notably, 5-ALA treatment alone slightly induced ROS production and sensitized the tumor to artemisinin toxicity in this setting. Because elevated intracellular ROS also induces widespread protein and DNA damage, we evaluated the amount of DNA double-strand breaks (DSBs) in sections of treated organoids. Indeed, we observed a strong increase in γH2AX (phosphorylated histone H2AX)-positive cells in 5-ALA and artemisinin-double-treated GFP-positive tumor tissue, but not in wild-type cells (Figures 3H, 9B, and 9C). Furthermore, the basal numbers of apoptotic cells (caspase 3-positive) and proliferating cells (Ki67-positive) were increased in GFP-positive tumor cells compared with normal tissue, but apoptosis was further increased in GFP-positive tumor cells upon combined treatment with artemisinin and 5-ALA (Figures 10A-C).

[0091] Because 5-ALA is clinically used for the diagnosis and treatment of human glioblastoma, we evaluated the combination of 5-ALA and artemisinin in an in vitro human glioblastoma-like tumor organoid model (Bian, S. et al. Genetically engineered cerebral organoids model brain tumor formation. Nat. Methods 15, 631-639 (2018)). These organoids were engineered to carry mutations in the tumor suppressor genes p53, NF1, and PTEN, which, along with GFP, allowed us to monitor tumor growth over time in the presence of artemisinin, 5-ALA, or a combination of both. While 5-ALA and artemisinin alone had little effect on the survival of non-transformed and transformed cells, the combination of both compounds specifically eliminated GFP-positive tumor cells in in vitro tumor models (Figures 4A and 11A). Quantification and imaging analysis in brain organoids confirmed the progressive loss of GFP-positive tumor cells and tissue, but not non-transformed cells (Figures 4B and 11C). These data indicate that 5-ALA and artemisinin induce increased tumor cell death in two highly aggressive brain tumor models through elevated ROS levels and increased DNA damage.

[0092] In summary, Applicant clearly demonstrates the requirement for porphyrin biosynthesis as a key endogenous pathway for artemisinin-induced cytotoxicity in eukaryotic cells. Using a high-throughput genetic screening system, Applicant identifies mitochondrial function, more specifically porphyrin / heme biosynthesis, required for the activity of this antimalarial and anticancer compound. Genetic and pharmacological modulation of porphyrin production was sufficient to modulate artemisinin toxicity and control artemisinin-induced reactive oxygen species levels in multiple cellular contexts and different species. Notably, induction of heme biosynthesis alone modestly elevated cellular ROS levels and sensitized cellular ROS to artemisinin-induced toxicity. Furthermore, Applicant demonstrates that combined treatment with 5-ALA and artemisinin can be used to specifically target brain cancer cells, particularly human glioblastoma, by inducing cell death in several independent in vitro human brain tumor organoid and spheroid model systems.

[0093] After demonstrating the potent synergistic effect of artemisinin (ART) and 5-aminolevulinic acid (5-ALA) for the treatment of brain cancer, Applicant next evaluated whether this synergistic effect extended to other ART derivatives, additional cancer types, and whether this dual combination synergized with currently applied cancer treatment regimens, particularly chemotherapy. Using 6-day viability analyses, Applicant confirmed that ART itself, and the most commonly used ART derivatives, dihydroartemisinin (DHA) and artesunate (ARS), in combination with 5-ALA, exhibited potent synergistic, antiproliferative effects against multiple glioblastoma cell lines and highly proliferative mouse embryonic stem cells (ESCs, Figures 12-1 to 12-6). From these data, Applicant derived cell type- and compound-specific dose combinations for the following analyses.

[0094] Glioblastoma multiforme is a highly aggressive cancer currently treated by maximal surgical resection, followed by radiation therapy, and concomitant and maintenance temozolomide (TMZ) therapy. We have shown that the combination of TMZ with 5-ALA, plus ART or DHA, is more effective than all other dual combinations in all glioblastoma lines tested (Figures 13-1 to 13-3). For example, in ESCs, only the dual combination of DHA and 5-ALA induces the cytotoxic effect of TMZ, demonstrating a clear synergistic effect.

[0095] These results extend to ARS, with the combination of ARS with 5-ALA and TMZ being more effective than replacing 5-ALA with its derivative, methyl-5-ALA (M-5-Ala), in four of the five glioblastoma cell lines tested (Figures 14-1 and 14-2).

[0096] Lomustine (CCNU) is an alternative to TMZ in the treatment of brain tumors, and as with TMZ, the cytostatic effect of lomustine (CCNU) can be particularly enhanced by combining it with ART and 5-ALA compared to all possible dual combinations (Figure 15).

[0097] Furthermore, we tested the combination of ART and its derivatives with 5-ALA in cancer cell lines derived from other tissues. We not only found a strong synergistic effect of ART / DHA / ARS and 5-ALA in reducing the survival rate of these cancer lines, but also found that this dual combination enhanced the efficacy of widely used chemotherapy drugs, including cisplatin (CP), carboplatin, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, vinorelbine, temozolomide, lomustine, and 5-fluorouracil (5-FU). This was specifically tested in two lung cancer cell lines (Figures 16-1 and 16-2), HepG2 cells (liver, Figure 17), MD-MBA-231 cells (breast, Figure 18), and MiaPaca-2 cells (pancreas, Figure 19). Taken together, our data demonstrate for the first time the strong potential of ART and ART derivative treatments in combination with 5-ALA and currently used chemotherapeutic agents, particularly anti-glioblastoma drugs, in cancers such as hematopoietic, brain, lung, liver, breast, and pancreatic cancers.

[0098] Therefore, the present invention provides a) artemisinin compound (1), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; and b) 5-aminolevulinic acid (2), methyl-5-aminolevulinic acid (2b), or a pharmaceutically acceptable salt or solvate thereof; and c) at least one chemotherapeutic agent, preferably one anti-glioblastoma drug Including, The present invention relates to a pharmaceutical composition for use in the prevention and / or treatment of hematopoietic cancer, brain cancer, pancreatic cancer, liver cancer, breast cancer and lung cancer, in particular glioblastoma.

[0099] Preferably, the present invention provides a method for producing a medicament for the treatment of a pulmonary arthritis. a) an artemisinin compound (1) selected from the group consisting of artemisinin (1a), dihydroartemisinin (1b), and artesunate (1e), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; and b) 5-aminolevulinic acid (2), methyl-5-aminolevulinic acid (2b), or a pharmaceutically acceptable salt or solvate thereof; and c) at least one chemotherapeutic agent, preferably one anti-glioblastoma drug Including, The present invention relates to a pharmaceutical composition for use in the prevention and / or treatment of hematopoietic cancer, brain cancer, pancreatic cancer, liver cancer, breast cancer and lung cancer, in particular glioblastoma.

[0100] Preferably, the present invention provides a method for producing a medicament for the treatment of a pulmonary arthritis. a) artemisinin compound (1), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; and b) 5-aminolevulinic acid (2), or a pharmaceutically acceptable salt or solvate thereof; and c) at least one chemotherapeutic agent, preferably one anti-glioblastoma drug Including, The present invention relates to a pharmaceutical composition for use in the prevention and / or treatment of hematopoietic cancer, brain cancer, pancreatic cancer, liver cancer, breast cancer and lung cancer, in particular glioblastoma.

[0101] Preferably, the present invention provides a method for producing a medicament for the treatment of a pulmonary arthritis. a) an artemisinin compound (1) selected from the group consisting of artemisinin (1a), dihydroartemisinin (1b), and artesunate (1e), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; and b) 5-aminolevulinic acid (2), or a pharmaceutically acceptable salt or solvate thereof; and c) at least one chemotherapeutic agent, preferably one anti-glioblastoma drug Including, The present invention relates to a pharmaceutical composition for use in the prevention and / or treatment of hematopoietic cancer, brain cancer, pancreatic cancer, liver cancer, breast cancer and lung cancer, in particular glioblastoma.

[0102] Even more preferably, the present invention relates to a pharmaceutical composition for use in the prevention and / or treatment of hematopoietic cancer, brain cancer, pancreatic cancer, liver cancer, breast cancer and lung cancer, in particular glioblastoma, comprising the following a to c: a) artemisinin compound (1), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; and b) 5-aminolevulinic acid (2), methyl-5-aminolevulinic acid (2b), or a pharmaceutically acceptable salt or solvate thereof; and c) temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevin, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, sertraline, irinotecan, clofazimine, and docetaxel; Preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevine, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, and sertraline; More preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, flubendazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, ribavirin, and vinorebin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, and doxorubicin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, and dactinomycin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and triptolide; Even more preferably, at least one anti-glioblastoma agent selected from the group comprising or consisting of temozolomide, lomustine, cisplatin, and 5-fluorouracil, most preferably temozolomide.

[0103] Even more preferably, the present invention relates to a pharmaceutical composition for use in the prevention and / or treatment of hematopoietic cancer, brain cancer, pancreatic cancer, liver cancer, breast cancer and lung cancer, in particular glioblastoma, comprising the following a to c: a) artemisinin compound (1), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; and b) 5-aminolevulinic acid (2), or a pharmaceutically acceptable salt or solvate thereof; and c) temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevin, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, sertraline, irinotecan, clofazimine, and docetaxel; Preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevine, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, and sertraline; More preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, flubendazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, ribavirin, and vinorebin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, and doxorubicin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, and dactinomycin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and triptolide; Even more preferably, at least one anti-glioblastoma agent selected from the group comprising or consisting of temozolomide, lomustine, cisplatin, and 5-fluorouracil, most preferably temozolomide.

[0104] Even more preferably, the present invention relates to a pharmaceutical composition for use in the prevention and / or treatment of hematopoietic cancer, brain cancer, pancreatic cancer, liver cancer, breast cancer and lung cancer, in particular glioblastoma, comprising the following a to c: a) an artemisinin compound (1) selected from the group consisting of artemisinin (1a), dihydroartemisinin (1b), and artesunate (1e), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; and b) 5-aminolevulinic acid (2), or a pharmaceutically acceptable salt or solvate thereof; and c) temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevin, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, sertraline, irinotecan, clofazimine, and docetaxel; Preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevine, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, and sertraline; More preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, flubendazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, ribavirin, and vinorebin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, and doxorubicin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, and dactinomycin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and triptolide; Even more preferably, at least one anti-glioblastoma agent selected from the group comprising or consisting of temozolomide, lomustine, cisplatin, and 5-fluorouracil, most preferably temozolomide.

[0105] For use in the prevention and / or treatment of cancer selected from hematopoietic cancer, brain cancer, pancreatic cancer, liver cancer, breast cancer, and lung cancer such as non-small cell lung cancer, preferably brain cancer, more preferably glioblastoma, the molar ratio of artemisinin compound to 5-aminolevulinic acid in the pharmaceutical composition is 1:5 to 1:5000, preferably 1:5 to 1:1000, more preferably 1:5 to 1:500, even more preferably 1:10 to 1:250, and most preferably 1:10 to 1:100.

[0106] For use in the prevention and / or treatment of cancer selected from hematopoietic cancer, brain cancer, pancreatic cancer, liver cancer, breast cancer, and lung cancer such as non-small cell lung cancer, preferably brain cancer, more preferably glioblastoma, the molar ratio of artemisinin compound to methyl-5-aminolevulinic acid in the pharmaceutical composition is 1:5 to 1:5000, preferably 1:5 to 1:1000, more preferably 1:5 to 1:500, even more preferably 1:10 to 1:250, and most preferably 1:10 to 1:100.

[0107] The pharmaceutical composition of the present invention comprises an artemisinin compound or a pharmaceutically acceptable salt, co-crystal, solvate thereof, and 5-aminolevulinic acid or methyl-5-aminolevulinic acid, preferably 5-aminolevulinic acid, as the only active ingredients, and one chemotherapeutic agent, preferably one anti-glioblastoma drug, in particular for the treatment of hematopoietic cancer, brain cancer, pancreatic cancer, liver cancer, breast cancer, and lung cancer such as non-small cell lung cancer, preferably brain cancer, more preferably glioblastoma.

[0108] In another aspect, the present invention relates to a method for the prevention and / or treatment of cancer selected from hematopoietic cancer, brain cancer, pancreatic cancer, liver cancer, breast cancer, and lung cancer, and in particular glioblastoma, the method comprising administering to a patient in need thereof a therapeutically effective amount of a pharmaceutical composition comprising: a) artemisinin compound (1), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; and b) 5-aminolevulinic acid (2), methyl-5-aminolevulinic acid, or a pharmaceutically acceptable salt or solvate thereof; and c) at least one chemotherapeutic agent, preferably one anti-glioblastoma drug.

[0109] Preferably, the present invention relates to a method for the prevention and / or treatment of cancer selected from hematopoietic cancer, brain cancer, pancreatic cancer, liver cancer, breast cancer, and lung cancer, in particular glioblastoma, the method comprising administering to a patient in need thereof a therapeutically effective amount of a pharmaceutical composition comprising the following a to c: a) an artemisinin compound (1) selected from the group consisting of artemisinin (1a), dihydroartemisinin (1b), and artesunate (1e), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; and b) 5-aminolevulinic acid (2), methyl-5-aminolevulinic acid, or a pharmaceutically acceptable salt or solvate thereof; and c) at least one chemotherapeutic agent, preferably one anti-glioblastoma drug.

[0110] Furthermore, the present invention relates to a method for preventing and / or treating cancer selected from hematopoietic cancer, brain cancer, pancreatic cancer, liver cancer, breast cancer, and lung cancer, particularly glioblastoma, which method comprises administering to a patient in need thereof a therapeutically effective amount of a pharmaceutical composition comprising the following a to c: a) artemisinin compound (1), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; and b) 5-aminolevulinic acid (2), or a pharmaceutically acceptable salt or solvate thereof; and c) at least one chemotherapeutic agent, preferably one anti-glioblastoma drug.

[0111] Furthermore, the present invention relates to a method for preventing and / or treating cancer selected from hematopoietic cancer, brain cancer, pancreatic cancer, liver cancer, breast cancer, and lung cancer, particularly glioblastoma, which method comprises administering to a patient in need thereof a therapeutically effective amount of a pharmaceutical composition comprising the following a to c: a) an artemisinin compound (1) selected from the group consisting of artemisinin (1a), dihydroartemisinin (1b), and artesunate (1e), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; and b) 5-aminolevulinic acid (2), or a pharmaceutically acceptable salt or solvate thereof; and c) at least one chemotherapeutic agent, preferably one anti-glioblastoma drug.

[0112] Even more preferably, the present invention relates to a method for the prevention and / or treatment of cancer selected from hematopoietic cancer, brain cancer, pancreatic cancer, liver cancer, breast cancer, and lung cancer, in particular glioblastoma, which method comprises administering to a patient in need thereof a therapeutically effective amount of a pharmaceutical composition comprising: a) artemisinin compound (1), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; and b) 5-aminolevulinic acid (2), methyl-5-aminolevulinic acid (2b), or a pharmaceutically acceptable salt or solvate thereof; and c) temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevin, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, sertraline, irinotecan, clofazimine, and docetaxel; Preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevine, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, and sertraline; More preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, flubendazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, ribavirin, and vinorebin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, and doxorubicin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, and dactinomycin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and triptolide; Even more preferably, at least one anti-glioblastoma agent selected from the group comprising or consisting of temozolomide, lomustine, cisplatin, and 5-fluorouracil, most preferably temozolomide.

[0113] Even more preferably, the present invention relates to a method for the prevention and / or treatment of cancer selected from hematopoietic cancer, brain cancer, pancreatic cancer, liver cancer, breast cancer, and lung cancer, in particular glioblastoma, which method comprises administering to a patient in need thereof a therapeutically effective amount of a pharmaceutical composition comprising: a) artemisinin compound (1), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; and b) 5-aminolevulinic acid (2), or a pharmaceutically acceptable salt or solvate thereof; and c) temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevin, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, sertraline, irinotecan, clofazimine, and docetaxel; Preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevine, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, and sertraline; More preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, flubendazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, ribavirin, and vinorebin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, and doxorubicin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, and dactinomycin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and triptolide; Even more preferably, at least one anti-glioblastoma agent selected from the group comprising or consisting of temozolomide, lomustine, cisplatin, and 5-fluorouracil, most preferably temozolomide.

[0114] Even more preferably, the present invention relates to a method for the prevention and / or treatment of cancer selected from hematopoietic cancer, brain cancer, pancreatic cancer, liver cancer, breast cancer, and lung cancer, in particular glioblastoma, which method comprises administering to a patient in need thereof a therapeutically effective amount of a pharmaceutical composition comprising: a) an artemisinin compound (1) selected from the group consisting of artemisinin (1a), dihydroartemisinin (1b), and artesunate (1e), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; and b) 5-aminolevulinic acid (2), or a pharmaceutically acceptable salt or solvate thereof; and c) temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevin, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, sertraline, irinotecan, clofazimine, and docetaxel; Preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevine, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, and sertraline; More preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, flubendazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, ribavirin, and vinorebin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, and doxorubicin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, and dactinomycin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and triptolide; Even more preferably, at least one anti-glioblastoma agent selected from the group comprising or consisting of temozolomide, lomustine, cisplatin, and 5-fluorouracil, most preferably temozolomide.

[0115] Optionally, the pharmaceutical composition of the present invention further comprises a pharmaceutically acceptable carrier, excipient and / or diluent for use in the prevention and / or treatment of cancer selected from hematopoietic cancer, brain cancer, pancreatic cancer, liver cancer, breast cancer, and lung cancer such as non-small cell lung cancer, preferably brain cancer, more preferably glioblastoma. The pharmaceutical composition of the present invention can be prepared at appropriate dosage levels in conventional solid or liquid carriers or diluents, and conventional pharmaceutically produced adjuvants, by known methods.

[0116] Pharmaceutical compositions of the present invention are typically administered in accordance with conventional pharmaceutical practice with a suitable acceptable carrier selected for oral administration according to the intended dosage form, i.e., in the form of tablets, capsules (either solid-filled, semi-solid-filled, or liquid-filled), powders for constitution, gels, elixirs, dispersible granules, syrups, suspensions, etc. For example, for oral administration in the form of tablets or capsules, the active drug component may be combined with any oral non-toxic pharmaceutically acceptable carrier, preferably an inert carrier such as lactose, starch, sucrose, cellulose, magnesium stearate, dicalcium phosphate, calcium sulfate, talc, mannitol, or ethyl alcohol (liquid-filled capsules). Additionally, suitable binders, lubricants, disintegrants, and coloring agents may be incorporated into the tablet or capsule. Powders and tablets may contain, as the active ingredient, from about 5 to about 95% by weight of the derivative according to general formula (I), or its analog compound, or a pharmaceutically active salt thereof.

[0117] Suitable binders include starch, gelatin, natural sugars, corn sweeteners, natural and synthetic gums such as acacia, sodium alginate, carboxymethylcellulose, polyethylene glycol, and waxes. Suitable lubricants may include boric acid, sodium benzoate, sodium acetate, sodium chloride, etc. Suitable disintegrants include starch, methylcellulose, guar gum, etc. Sweeteners and flavorings, as well as preservatives, may also be included, where appropriate. Disintegrants, diluents, lubricants, binders, etc. are described in more detail below.

[0118] Furthermore, the pharmaceutical compositions of the present invention can be formulated in sustained release forms that provide rate-controlled release of any one or more components or active ingredients to optimize therapeutic efficacy, such as anti-cancer activity or activity against cancer metastasis. Suitable dosage forms for sustained release include tablets with different disintegration rate or controlled release layers, polymer matrices impregnated with the active ingredient and formed into a tablet, or capsules containing such impregnated or encapsulated porous polymer matrices.

[0119] Liquid preparations include solutions, suspensions, and emulsions. For example, water or water / propylene glycol solutions for parenteral injection, or the addition of sweeteners and opacifiers for oral solutions, suspensions, and emulsions may be mentioned. Liquid preparations may also include solutions for intranasal administration.

[0120] Aerosol preparations suitable for inhalation may include solutions and solids in powder form, which may be in combination with a pharmaceutically acceptable carrier, such as an inert compressed gas, e.g., nitrogen.

[0121] For preparing suppositories, a low melting wax, such as a mixture of fatty acid glycerides like cocoa butter, is first melted, and the active ingredient is then dispersed homogeneously therein by, for example, stirring. The molten homogeneous mixture is then poured into convenient sized molds and allowed to cool and solidify.

[0122] Also included are solid preparations which are intended to be converted, shortly before use, to liquid preparations for either oral or parenteral administration. Such liquid forms include solutions, suspensions, and emulsions.

[0123] The term "capsule" as used herein refers to a specific container or enclosure made of, for example, methylcellulose, polyvinyl alcohol, or modified gelatin or starch, for holding or containing a composition including an active ingredient. Hard-shell capsules are typically made from a blend of relatively high gel strength gelatin from bone or pigskin. The capsule itself may contain small amounts of dyes, opacifiers, plasticizers, and / or preservatives.

[0124] A tablet is understood to be a compressed or molded solid dosage form containing the active ingredient together with suitable diluents. Tablets may be prepared by compression of mixtures or granulations obtained by wet granulation, dry granulation, or by compression methods well known to those skilled in the art.

[0125] Oral gel refers to the active ingredients dispersed or solubilized in a hydrophilic semi-solid matrix.

[0126] Powders for constitution refer to a powder mixture containing the active ingredients and suitable diluents, which can be suspended, for example, in water or juice.

[0127] Suitable diluents are usually substances that make up the majority of the composition or dosage form.Suitable diluents include, for example, sugars such as lactose, sucrose, mannitol, and sorbitol, starches derived from wheat, corn, rice, and potato, and celluloses such as microcrystalline cellulose.The amount of diluent in the composition can range from about 5% to about 95% by weight, preferably from about 25% to about 75% by weight, and more preferably from about 30% to about 60% by weight of the total composition.

[0128] The term disintegrant refers to a material added to a composition to help break apart (disintegrate) and release the pharmaceutically active ingredients in a drug. Suitable disintegrants include starch, "cold water soluble" modified starches such as sodium carboxymethyl starch, natural and synthetic gums such as locust bean, karaya, guar, tragacanth, and agar, cellulose derivatives such as methylcellulose and sodium carboxymethylcellulose, microcrystalline cellulose and cross-linked microcrystalline cellulose such as cross-caramellose sodium, alginic acid and alginates such as sodium alginate, clays such as bentonite, and effervescent mixtures. The amount of disintegrant in the composition can range from about 2% to about 20% by weight of the composition, more preferably from about 5% to about 10% by weight.

[0129] Binders are substances that bind or "glue" powder particles together, holding them together by forming granules and acting as the "adhesive" in a formulation. Binders add cohesive strength already available in the diluent or filler. Suitable binders include sugars such as sucrose; starches derived from wheat, corn, rice, and potato; natural gums such as acacia, gelatin, and tragacanth; seaweed derivatives such as alginic acid, sodium alginate, and calcium ammonium alginate; cellulosic materials such as methylcellulose, sodium carboxymethylcellulose, and hydroxypropylmethylcellulose; polyvinylpyrrolidone; and inorganic compounds such as magnesium aluminum silicate. The amount of binder in the composition may range from about 2% to about 20% by weight of the composition, preferably from about 3% to about 10%, and more preferably from about 3% to about 6%.

[0130] Lubricants are a class of substances added to dosage forms to reduce friction or wear, allowing tablet granules and the like to be released from the die after compression. Suitable lubricants include metal stearates such as magnesium stearate, calcium stearate, or potassium stearate; stearic acid; high-melting-point waxes; and other water-soluble lubricants such as sodium chloride, sodium benzoate, sodium acetate, sodium oleate, polyethylene glycol, and D,L-leucine. Because lubricants must be present on the surface of the granules, they are usually added at the very last step before compression. The amount of lubricant in the composition can range from about 0.2% to about 5% by weight of the composition, preferably from about 0.5% to about 2% by weight, and more preferably from about 0.3% to about 1.5% by weight of the composition.

[0131] Glidants are materials that prevent caking in the components of pharmaceutical compositions and improve the flow properties of granules so that they flow smoothly and uniformly. Suitable glidants include silicon dioxide and talc. The amount of glidant in the composition can range from about 0.1% to about 5% by weight, preferably from about 0.5% to about 2% by weight of the final composition.

[0132] A colorant is an excipient that imparts color to a composition or dosage form. Such excipients can include food-grade dyes adsorbed onto a suitable adsorbent, such as clay or aluminum oxide. The amount of colorant can vary from about 0.1% to about 5% by weight of the composition, preferably from about 0.1% to 1% by weight.

[0133] Therefore, the present invention provides a) artemisinin compound (1), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; and b) 5-aminolevulinic acid (2), methyl-5-aminolevulinic acid (2b), or a pharmaceutically acceptable salt or solvate thereof; and c) at least one chemotherapeutic agent, preferably an anti-glioblastoma drug; and d) Pharmaceutically acceptable carriers, excipients, and / or diluents The present invention provides a pharmaceutical composition comprising or consisting of:

[0134] Furthermore, the present application preferably a) an artemisinin compound (1) selected from the group consisting of artemisinin (1a), dihydroartemisinin (1b), and artesunate (1e), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; b) 5-aminolevulinic acid (2), methyl-5-aminolevulinic acid (2b), or a pharmaceutically acceptable salt or solvate thereof; and c) at least one chemotherapeutic agent, preferably an anti-glioblastoma drug; and d) Pharmaceutically acceptable carriers, excipients, and / or diluents The present invention relates to a pharmaceutical composition comprising or consisting of:

[0135] More preferably, the present application provides a) artemisinin (1a), or artesunate (1e), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; b) 5-aminolevulinic acid (2), methyl-5-aminolevulinic acid (2b), or a pharmaceutically acceptable salt or solvate thereof; and c) at least one chemotherapeutic agent, preferably an anti-glioblastoma drug; and d) Pharmaceutically acceptable carriers, excipients, and / or diluents The present invention relates to a pharmaceutical composition comprising or consisting of:

[0136] Even more preferably, the present application provides a) artesunate (1e), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; b) 5-aminolevulinic acid (2), methyl-5-aminolevulinic acid (2b), or a pharmaceutically acceptable salt or solvate thereof; and c) at least one chemotherapeutic agent, preferably an anti-glioblastoma drug; and d) Pharmaceutically acceptable carriers, excipients, and / or diluents The present invention relates to a pharmaceutical composition comprising or consisting of:

[0137] Even more preferably, the present application provides a) artemisinin (1a), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; b) 5-aminolevulinic acid (2), methyl-5-aminolevulinic acid (2b), or a pharmaceutically acceptable salt or solvate thereof; and c) at least one chemotherapeutic agent, preferably an anti-glioblastoma drug; and d) Pharmaceutically acceptable carriers, excipients, and / or diluents The present invention relates to a pharmaceutical composition comprising or consisting of:

[0138] Furthermore, the present invention provides a) artemisinin compound (1), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; and b) 5-aminolevulinic acid (2), or a pharmaceutically acceptable salt or solvate thereof; and c) at least one chemotherapeutic agent, preferably an anti-glioblastoma drug; and d) Pharmaceutically acceptable carriers, excipients, and / or diluents The present invention provides a pharmaceutical composition comprising or consisting of:

[0139] Furthermore, the present application preferably a) an artemisinin compound (1) selected from the group consisting of artemisinin (1a), dihydroartemisinin (1b), and artesunate (1e), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; and b) 5-aminolevulinic acid (2), or a pharmaceutically acceptable salt or solvate thereof; and c) at least one chemotherapeutic agent, preferably an anti-glioblastoma drug; and d) Pharmaceutically acceptable carriers, excipients, and / or diluents The present invention relates to a pharmaceutical composition comprising or consisting of:

[0140] More preferably, the present application provides a) artemisinin (1a), or artesunate (1e), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; and b) 5-aminolevulinic acid (2), or a pharmaceutically acceptable salt or solvate thereof; and c) at least one chemotherapeutic agent, preferably an anti-glioblastoma drug; and d) Pharmaceutically acceptable carriers, excipients, and / or diluents The present invention relates to a pharmaceutical composition comprising or consisting of:

[0141] Even more preferably, the present application provides a) artesunate (1e), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; and b) 5-aminolevulinic acid (2), or a pharmaceutically acceptable salt or solvate thereof; and c) at least one chemotherapeutic agent, preferably an anti-glioblastoma drug; and d) Pharmaceutically acceptable carriers, excipients, and / or diluents The present invention relates to a pharmaceutical composition comprising or consisting of:

[0142] Even more preferably, the present application provides a) artemisinin (1a), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; and b) 5-aminolevulinic acid (2), or a pharmaceutically acceptable salt or solvate thereof; and c) at least one chemotherapeutic agent, preferably an anti-glioblastoma drug; and d) Pharmaceutically acceptable carriers, excipients, and / or diluents The present invention relates to a pharmaceutical composition comprising or consisting of:

[0143] The present invention further provides a pharmaceutical composition comprising or consisting of the following a) to d): a) artemisinin compound (1), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; and b) 5-aminolevulinic acid (2), methyl-5-aminolevulinic acid (2b), or a pharmaceutically acceptable salt or solvate thereof; and c) temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevin, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, sertraline, irinotecan, clofazimine, and docetaxel; Preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevine, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, and sertraline; More preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, flubendazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, ribavirin, and vinorebin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, and doxorubicin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, and dactinomycin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and triptolide; Even more preferably, at least one anti-glioblastoma drug selected from the group comprising or consisting of temozolomide, lomustine, cisplatin, and 5-fluorouracil; Most preferably, temozolomide; and d) Pharmaceutically acceptable carriers, excipients, and / or diluents.

[0144] The present application further provides a pharmaceutical composition preferably comprising or consisting of the following a) to d): a) an artemisinin compound (1) selected from the group consisting of artemisinin (1a), dihydroartemisinin (1b), and artesunate (1e), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; b) 5-aminolevulinic acid (2), methyl-5-aminolevulinic acid (2b), or a pharmaceutically acceptable salt or solvate thereof; and c) temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevin, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, sertraline, irinotecan, clofazimine, and docetaxel; Preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevine, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, and sertraline; More preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, flubendazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, ribavirin, and vinorebin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, and doxorubicin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, and dactinomycin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and triptolide; Even more preferably, at least one anti-glioblastoma drug selected from the group comprising or consisting of temozolomide, lomustine, cisplatin, and 5-fluorouracil; Most preferably, temozolomide; and d) Pharmaceutically acceptable carriers, excipients, and / or diluents.

[0145] More preferably, the present application provides a pharmaceutical composition comprising or consisting of the following a to d: a) artemisinin (1a), or artesunate (1e), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; b) 5-aminolevulinic acid (2), methyl-5-aminolevulinic acid (2b), or a pharmaceutically acceptable salt or solvate thereof; and c) temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevin, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, sertraline, irinotecan, clofazimine, and docetaxel; Preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevine, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, and sertraline; More preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, flubendazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, ribavirin, and vinorebin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, and doxorubicin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, and dactinomycin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and triptolide; Even more preferably, at least one anti-glioblastoma drug selected from the group comprising or consisting of temozolomide, lomustine, cisplatin, and 5-fluorouracil; Most preferably, temozolomide; and d) Pharmaceutically acceptable carriers, excipients, and / or diluents.

[0146] Even more preferably, the present application provides a pharmaceutical composition comprising or consisting of: a) artesunate (1e), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; b) 5-aminolevulinic acid (2), methyl-5-aminolevulinic acid (2b), or a pharmaceutically acceptable salt or solvate thereof; and c) temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevin, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, sertraline, irinotecan, clofazimine, and docetaxel; Preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevine, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, and sertraline; More preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, flubendazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, ribavirin, and vinorebin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, and doxorubicin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, and dactinomycin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and triptolide; Even more preferably, at least one anti-glioblastoma drug selected from the group comprising or consisting of temozolomide, lomustine, cisplatin, and 5-fluorouracil; Most preferably, temozolomide; and d) Pharmaceutically acceptable carriers, excipients, and / or diluents.

[0147] Even more preferably, the present application provides a pharmaceutical composition comprising or consisting of: a) artemisinin (1a), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; b) 5-aminolevulinic acid (2), methyl-5-aminolevulinic acid (2b), or a pharmaceutically acceptable salt or solvate thereof; and c) temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevin, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, sertraline, irinotecan, clofazimine, and docetaxel; Preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevine, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, and sertraline; More preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, flubendazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, ribavirin, and vinorebin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, and doxorubicin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, and dactinomycin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and triptolide; Even more preferably, at least one anti-glioblastoma drug selected from the group comprising or consisting of temozolomide, lomustine, cisplatin, and 5-fluorouracil; Most preferably, temozolomide; and d) Pharmaceutically acceptable carriers, excipients, and / or diluents.

[0148] Preferably, the present invention provides a pharmaceutical composition comprising or consisting of the following a to d: a) artemisinin compound (1), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; and b) 5-aminolevulinic acid (2), or a pharmaceutically acceptable salt or solvate thereof; and c) temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevin, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, sertraline, irinotecan, clofazimine, and docetaxel; Preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevine, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, and sertraline; More preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, flubendazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, ribavirin, and vinorebin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, and doxorubicin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, and dactinomycin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and triptolide; Even more preferably, at least one anti-glioblastoma drug selected from the group comprising or consisting of temozolomide, lomustine, cisplatin, and 5-fluorouracil; Most preferably, temozolomide; and d) Pharmaceutically acceptable carriers, excipients, and / or diluents.

[0149] The present application further provides a pharmaceutical composition preferably comprising or consisting of the following a) to d): a) an artemisinin compound (1) selected from the group consisting of artemisinin (1a), dihydroartemisinin (1b), and artesunate (1e), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; b) 5-aminolevulinic acid (2), or a pharmaceutically acceptable salt or solvate thereof; and c) temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevin, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, sertraline, irinotecan, clofazimine, and docetaxel; Preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevine, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, and sertraline; More preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, flubendazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, ribavirin, and vinorebin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, and doxorubicin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, and dactinomycin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and triptolide; Even more preferably, at least one anti-glioblastoma drug selected from the group comprising or consisting of temozolomide, lomustine, cisplatin, and 5-fluorouracil; Most preferably, temozolomide; and d) Pharmaceutically acceptable carriers, excipients, and / or diluents.

[0150] More preferably, the present application provides a pharmaceutical composition comprising or consisting of the following a to d: a) artemisinin (1a), or artesunate (1e), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; b) 5-aminolevulinic acid (2), or a pharmaceutically acceptable salt or solvate thereof; and c) temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevin, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, sertraline, irinotecan, clofazimine, and docetaxel; Preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevine, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, and sertraline; More preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, flubendazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, ribavirin, and vinorebin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, and doxorubicin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, and dactinomycin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and triptolide; Even more preferably, at least one anti-glioblastoma drug selected from the group comprising or consisting of temozolomide, lomustine, cisplatin, and 5-fluorouracil; Most preferably, temozolomide; and d) Pharmaceutically acceptable carriers, excipients, and / or diluents.

[0151] Even more preferably, the present application provides a pharmaceutical composition comprising or consisting of: a) artesunate (1e), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; b) 5-aminolevulinic acid (2), or a pharmaceutically acceptable salt or solvate thereof; and c) temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevin, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, sertraline, irinotecan, clofazimine, and docetaxel; Preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevine, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, and sertraline; More preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, flubendazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, ribavirin, and vinorebin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, and doxorubicin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, and dactinomycin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and triptolide; Even more preferably, at least one anti-glioblastoma drug selected from the group comprising or consisting of temozolomide, lomustine, cisplatin, and 5-fluorouracil; Most preferably, temozolomide; and d) Pharmaceutically acceptable carriers, excipients, and / or diluents.

[0152] Even more preferably, the present application provides a pharmaceutical composition comprising or consisting of: a) artemisinin (1a), or a pharmaceutically acceptable salt, co-crystal, or solvate thereof; b) 5-aminolevulinic acid (2), or a pharmaceutically acceptable salt or solvate thereof; and c) temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevin, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, sertraline, irinotecan, clofazimine, and docetaxel; Preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, vincristine, vinorevine, nisoldipine, deoxyadenosine, chloro-2'-deoxyadenosine, 5-nonyloxytryptamine, 2(1H)-pyrimidinone, pitavastatin, and sertraline; More preferably, temozolomide, dexamethasone, lomustine, methotrexate, everolimus, carmustine, cyclophosphamide, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, topotecan, flubendazole, itraconazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, idarubicin, ribavirin, flubendazole, vindesine sulfate, cerivastatin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, ribavirin, and vinorebin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, and vinorevin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, dactinomycin, and doxorubicin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, homoharringtonine, and dactinomycin; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, triptolide, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and homoharringtonine; Even more preferably, temozolomide, lomustine, cisplatin, carboplatin, 5-fluorouracil, and triptolide; Even more preferably, at least one anti-glioblastoma drug selected from the group comprising or consisting of temozolomide, lomustine, cisplatin, and 5-fluorouracil; Most preferably, temozolomide; and d) Pharmaceutically acceptable carriers, excipients, and / or diluents.

[0153] The pharmaceutical compositions of the present invention may be administered in combination with radiation therapy, immunotherapy, electromagnetic field therapy, hyperthermia, chemotherapy, cancer immunotherapy and / or small molecule-based therapy.

[0154] In some embodiments, pharmaceutical compositions of the present invention are used, preferably in combination with only one chemotherapeutic agent, preferably only one anti-glioblastoma agent, to prevent and / or treat hematopoietic cancer, brain cancer, pancreatic cancer, liver cancer, breast cancer, and lung cancer, such as non-small cell lung cancer, preferably brain cancer, more preferably glioblastoma, wherein the only chemotherapeutic agent, preferably the only anti-glioblastoma agent, is selected from temozolomide.

[0155] As used herein, "combination" or "pharmaceutical combination" means the product resulting from the mixing or combining of two or more active ingredients and includes both fixed and non-fixed combinations of the active ingredients.

[0156] The term "fixed combination" or "fixed dose" means that the active ingredients, e.g., a compound of Formula 1, and a combination partner, i.e., an anticancer agent, are both administered to a patient simultaneously in the form of a single entity or dosage. In other words: the active ingredients are present in one dosage form, e.g., one tablet or one capsule.

[0157] The term "non-fixed combination" means that the active ingredients, e.g., a compound of Formula 1, and a combination partner, i.e., an anti-cancer agent, are administered to a patient together as separate entities either simultaneously, concurrently, or sequentially without any specific time limit, where said administration provides therapeutically effective levels of the two compounds in the body of a mammal or human in need thereof. The latter also applies to cocktail therapy, e.g., the administration of three or more anti-cancer agents.

[0158] Said pharmaceutical combinations may be administered simultaneously or separately, independently, within time intervals, particularly here within these time intervals which allow the combination partners to exhibit a synergistic effect.

[0159] The term "synergistic effect" means that the therapeutic effect observed after administration of two or more active ingredients (e.g., an artemisinin compound, 5-ALA, at least one chemotherapeutic agent, or preferably at least one anti-glioblastoma drug) is greater than the effect obtained by administering each active ingredient alone. Thus, a synergistic effect exists when the effect obtained is greater than the merely additive effect expected from simultaneous administration of the active ingredients.

[0160] "Synergistic increase" means that the combination of two or more active ingredients (e.g., an artemisinin compound, 5-ALA, and at least one anti-glioblastoma drug) results in an increase in cancer cell death beyond the merely additive effect expected by co-administration of the active ingredients.

[0161] "Synergistic reduction" means that the combination of two or more active ingredients (e.g., one artemisinin compound, 5-ALA, at least one anti-glioblastoma drug) results in a reduction in one or more symptoms of cancer that exceeds the merely additive effect expected by co-administration of the active ingredients.

[0162] In other examples of synergy, a therapeutic effect is observed for the combination of two or more active ingredients, where one or more active ingredients are present at doses that are normally non-therapeutic. In other examples of synergy, the combination of two or more active ingredients results in an unexpected reduction in toxicity (i.e., a toxicity level that is lower than the sum of the toxicities observed after administration of the single agents).

[0163] However, synergistic effects may also be obtained by combining the administration of the pharmaceutical composition of the present invention with treatment with radiation therapy, immunotherapy, electromagnetic field therapy, hyperthermia, chemotherapy, cancer immunotherapy, or any other small molecule-based therapy as outlined above.

[0164] In some embodiments, the pharmaceutical composition is used in the form of a tablet, capsule, syrup, solution, suspension, emulsion, or gel.

[0165] In some embodiments, the pharmaceutical composition is administered orally or parenterally for the prevention and / or treatment of hematopoietic cancer, brain cancer, pancreatic cancer, liver cancer, breast cancer, and lung cancer such as non-small cell lung cancer, preferably brain cancer, more preferably glioblastoma.

[0166] Preferred formulations are suitable for oral application. These dosage forms include, for example, pills, tablets, film tablets, coated tablets, capsules, powders and deposits.

[0167] Parenteral applications include dermal, intradermal, intragastric, intracutaneous, intravascular, intravenous, intramuscular, intraperitoneal, intranasal, intravaginal, buccal, percutan, rectal, subcutaneous, sublingual, topical, or transdermal applications. Preferably, parenteral applications include intradermal, intragastric, intracutaneous, intravascular, intravenous, intramuscular, subcutaneous, sublingual, topical, transdermal, and inhalation.

[0168] The transdermal compositions may take the form of creams, lotions, aerosols and / or emulsions and may be included in a transdermal patch of the matrix or reservoir type as are known in the art for this purpose.

[0169] The term "capsule" as used herein refers to a specific container or enclosure made of, for example, methylcellulose, polyvinyl alcohol, modified gelatin, or starch, for holding or containing a composition containing an active ingredient. Hard-shell capsules are typically made from a blend of relatively high-gel-strength gelatin, typically from bone or pigskin. The capsule itself may contain small amounts of dyes, opacifiers, plasticizers, and / or preservatives. A tablet is understood to be a compressed or molded solid dosage form containing the active ingredient together with suitable diluents. Tablets may be prepared by compressing mixtures or granulations obtained by wet granulation, dry granulation, or compression methods well known to those skilled in the art.

[0170] Oral gel refers to the active ingredient dispersed or solubilized in a hydrophilic semi-solid matrix. Powder for constitution refers to a powder mixture containing the active ingredient and suitable diluents, which can be suspended, for example, in water or juice.

[0171] Additionally, the pharmaceutical compositions of the present invention may be formulated in sustained release form to provide rate-controlled release of any one or more components or active ingredients to optimize therapeutic efficacy, e.g., anti-cancer activity, etc. Suitable dosage forms for sustained release include tablets with different disintegration rate or controlled release layers, polymer matrices impregnated with the active ingredient and formed into a tablet, or capsules containing such impregnated or encapsulated porous polymer matrices.

[0172] In some embodiments, for the prevention and / or treatment of glioblastoma, the artemisinin compound, particularly artemisinin (1a), contained in the pharmaceutical composition is administered in the range of 0.01 to 100 mg / kg, preferably 0.1 to 100 mg / kg, more preferably 0.1 to 50 mg / kg, and most preferably 0.1 to 15 mg / kg of body weight per day, and 5-aminolevulinic acid is administered in the range of 0.01 to 200 mg / kg, preferably 0.1 to 200 mg / kg, more preferably 0.1 to 50 mg / kg, and most preferably 1 to 50 mg / kg of body weight per day.

[0173] Preferably, the dosage of the present invention for artemisinin compounds, particularly artemisinin, is at least 0.10, 0.15, 0.20, 0.25, 0.30, 0.35, 0.40, 0.45, 0.50, 0.55, 0.60, 0.65, 0.70, 0.75, 0.80, 0.85, 0.90, 0.95, 1.00, 1.05, 1.10, 1.15, 1.20, 1.25, 1.30, 1.40, 1.50, 1.60, 1.70, 1.80, 1.90, 2.00, 2.10, 2.25, 2.30, 2.40, 2.50, 2.60, 2.70, 2.80, 2.90, 3.00, 3.10, 3.20, 3.30, 3.40, 3.50, 3.60, 3.70, 3.80, 3.90, 4.00, 4.10, 4.20, 4.30, 4.40, 4.50, 4.50, 4.60, 4.7 ... , 1.30, 1.35, 1.40, 1.45, 1.50, 2.00, 2.50, 3.00, 3.50, 4.00, 4.50, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0, 10.5, 11.0, 11.5, 12.0, 12.5, 13.0, 13.5, 14.0, 14.5, or 15.0 mg / kg of artemisinin.

[0174] Preferably, the dosage of the present invention for 5-aminolevulinic acid is at least 0.10, 0.15, 0.20, 0.25, 0.30, 0.35, 0.40, 0.45, 0.50, 0.55, 0.60, 0.65, 0.70, 0.75, 0.80, 0.85, 0.90, 0.95, 1.00, 1.05, 1.10, 1.15, 1.20, 1.25, 1.30, 1.35, 1.40, 1.45, 1.50, 2.00, 2.50, 3.00, 3.50, 4.00, 4.50, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0, 10.5, 11.0, 11.5, 12.0, 12.5, 13.0, 13.5, 14.0, 14.5, 15.0, 17.0, 20.0, 25.0, 30.0, 35.0, 40.0, 45.0, or 50.0 mg / kg of 5-aminolevulinic acid.

[0175] In some embodiments, the pharmaceutical composition is used for the prevention and / or treatment of glioblastoma, preferably in combination with at least one chemotherapeutic agent, preferably at least one anti-glioblastoma agent, wherein the at least one anti-glioblastoma agent is administered in the range of 0.01 to 100 mg / kg of body weight per day.

[0176] Preferably, the dose of the present invention for a chemotherapeutic agent, preferably an anti-glioblastoma agent, is at least 0.10, 0.15, 0.20, 0.25, 0.30, 0.35, 0.40, 0.45, 0.50, 0.55, 0.60, 0.65, 0.70, 0.75, 0.80, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, 2000, 2100, 2200, 2300, 2400, 2500, 2600, 2700, 2800, 2900, 3000, 3100, 3200, 3300, 3400, 3500, 3600, 3700, 3800, 3900, 4000, 4100, 4200, 4300, 4400, 4500, 4600, 4700, 4800, 4900, 5100, 5200, 5300, 5400, 5500, 5600, 5700, 5800, 5900, 6000, 6100, 6200, 6300, 6400, 6500, 6600, 6700, 6800, 6900, 7000, 7100, 7200, 7300, 7400, 7500, 0.85, 0.90, 0.95, 1.00, 1.05, 1.10, 1.15, 1.20, 1.25, 1.30, 1.35, 1.40, 1.45, 1.50, 2.00, 2.50, 3.00, 3.50, 4.00, 4.50, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0, 10.5, 11.0, 11.5, 12.0, 12.5, 13.0, 13.5, 14.0, 14.5 or 15.0 mg / kg of chemotherapy or anti-glioblastoma drug, respectively.

[0177] Examples of such pharmaceutical compositions include, but are not limited to, solutions for injection, dry products that can be dissolved or suspended in a pharmaceutically acceptable vehicle for injection, suspensions for injection, and emulsions.

[0178] Some suitable vehicles that can be used to provide pharmaceutical compositions for parenteral administration include, but are not limited to, water for injection, USP; aqueous vehicles such as, but not limited to, sodium chloride injection, Ringer's injection, dextrose injection, dextrose and sodium chloride injection, and lactated Ringer's injection; water-miscible vehicles such as, but not limited to, ethyl alcohol, polyethylene glycol, and polypropylene glycol; and non-aqueous vehicles such as, but not limited to, corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate, isopropyl myristate, and benzyl benzoate.

[0179] The appropriate dosage of active ingredient can be determined by a skilled physician.The actual dosage level of active ingredient can be varied to obtain the amount of active ingredient that is effective for achieving the desired therapeutic response for a specific patient, composition, and administration method without causing toxicity to the patient.Therefore, the dosage is typically an effective or therapeutically effective dosage.

[0180] The selected dosage level will depend on various pharmacokinetic factors, including the activity of the particular composition of the invention being used, the route of administration, the time of administration, the excretion rate of the particular compound being used, the duration of treatment, other drugs, compounds and / or materials being used in combination with the particular composition being used, the age, sex, weight, condition, general health and previous medical history of the patient being treated, and similar factors well known in the medical arts.

[0181] Dosage regimen can be adjusted to provide the optimum desired response. For example, it can be administered in a single dose (e.g., a single daily dose), in divided doses over time, or the dose can be proportionally reduced or increased according to the exigencies of the therapeutic situation. As used herein, dosage unit form refers to physically discrete units suitable as unitary dosages for the patient to be treated; each unit contains a predetermined amount of active compound calculated to produce the desired therapeutic effect, in association with the required pharmaceutical carrier.

[0182] In the compositions and products of the present invention, the active ingredients may each be present in a concentration of, for example, 0.001 to 20% by weight, preferably 0.01 to 10% by weight, more preferably 0.02 to 5% by weight, and even more preferably 1 to 4% by weight, based on the total weight of the composition or product. In certain embodiments, each of the three active ingredients is present in a concentration of 1 to 3% by weight.

[0183] In one currently preferred embodiment, the artemisinin compound, particularly artemisinin, is formulated for daily administration at 50-500 mg (more preferably, 100-300 mg, such as about 200 mg) (based on a body weight of about 70 kg; dosage can be adjusted proportionally to body weight). Preferably, the artemisinin is formulated for oral or parenteral administration.

[0184] Preferably, the artemisinin compound, particularly artemisinin, is administered for a period of 3 to 30 days, more preferably 10 to 20 days, e.g., about 14 days. This period may correspond to a treatment cycle in a dosing regimen including multiple such treatment cycles (e.g., at least 2, 3, 4, 5, 6, or more cycles, e.g., with such cycles continuing until a desired therapeutic result is achieved). Each treatment cycle may be separated by a break in artemisinin compound administration, particularly artemisinin administration; such a break may allow, for example, bone marrow recovery. The break in administration may include not administering the artemisinin compound, particularly artemisinin, for a period of 3 to 14 days, more preferably 5 to 10 days, e.g., about 7 days.

[0185] In one exemplary such embodiment, the artemisinin compound, particularly artemisinin, is administered at about 200 mg daily for two-week treatment cycles, each cycle being followed by a one-week break.

[0186] In some embodiments, the pharmaceutical composition is used for the prevention and / or treatment of glioblastoma, preferably in combination with radiation therapy, immunotherapy, electromagnetic field therapy, and / or hyperthermia.

[0187] In a preferred embodiment, the pharmaceutical composition is used for the prevention and / or treatment of glioblastoma, wherein the glioblastoma is selected from proneural (PN), mesenchymal (MES), and classical (CL) glioblastoma subtypes. [Brief explanation of the drawings]

[0188] [Figure 1-1] Figure 1: Haploid screening in mouse stem cells delineates porphyrin biosynthesis as a prerequisite for artemisinin toxicity in mammalian cells. Figure 1-1A: Schematic of haploid embryonic stem cell screening for compound target identification. Figure 1-1B: Porphyrin biosynthetic pathway. The subcellular location (cytosol, mitochondria) of key enzymes (bold, in capital letters) and cofactors (standard, in capital letters) in the screen (retroviral mutagenesis) and loss-of-function scores (italics) are shown. [Figure 1-2]Figure 1: Haploid screening in mouse stem cells delineates porphyrin biosynthesis as a prerequisite for artemisinin toxicity in mammalian cells. Figure 1-2C: GO term analysis reveals porphyrin biosynthesis as the primary pathway targeted by artemisinin. Figure 1-2D: Competitive growth analysis of artemisinin-resistant single-cell clones. Wild-type (mCherry+_Cre) and knockout (GFP+) sister clones derived from mutant (retrovirus-intronic RE, darker shading; Tol2 transposon-intronic T2, lighter shading) resistant colonies were treated with artemisinin and analyzed using flow cytometry and Sanger sequencing for insertion site mapping. [Figure 2-1] Figure 2: Modulation of porphyrin biosynthesis is sufficient to alter artemisinin toxicity in cells via ROS generation. Figure 2-1A,B: Cell viability of mouse embryonic stem cells treated with artemisinin in combination with (A) the Ppox inhibitor acifluorfen or (B) 5-ALA (0.5 mM). Alamar Blue staining was used to assess viability after 48 hours of treatment. Figure 2-1C: Cell viability of artemisinin-treated mouse breast cancer cells (4T1) in the presence and absence of 5-ALA (0.5 mM). Alamar Blue was used to determine cell viability after 48 hours of treatment. Figure 2-1D: Survival of artemisinin-treated primary human glioblastoma cells from a cancer patient in response to 5-ALA administration. Viability was assessed using CellTiter-Glo after 72 hours. Experiments were performed in triplicate. Values are means ± SD. [Figure 2-2]Figure 2: Modulation of porphyrin biosynthesis is sufficient to alter artemisinin toxicity in cells via ROS generation. Figure 2E,F: (E) ROS levels (DHE staining, PE582 / 15nm-MFI) and (F) cell viability in neuroblastoma cells (SHSY5Y) treated with artemisinin (0.5 μM), 5-ALA (0.25 mM), or Ppox inhibitor (10 μM). DHE fluorescence (ROS levels) and cell number (viability) were assessed by flow cytometry and automated cell counting. Experiments were performed twice in triplicate. Values represent the mean ± SD. [Figure 3-1] Figure 3: Brain tumor organoids (CNS-PNET) show increased sensitivity to artemisinin and 5-ALA combination therapy. Figure 3-1A: Human brain primitive neuroectodermal tumor organoids for compound profiling. Figure 3-1B: Schematic workflow diagram and analysis of 5-ALA and artemisinin-treated organoids. Figure 3-1C,D: (C) Representative fluorescent and (D) brightfield images of tumor organoids (CNS-PNET) treated with DMSO (control), 5-ALA (0.0625 mM), artemisinin (1 μM), or 5-ALA and artemisinin (0.0625 mM and 1 μM). Organoids were imaged on days 1, 5, and 7. Scale bar 500 μm. [Figure 3-2]Figure 3: Brain tumor organoids (CNS-PNET) show increased sensitivity to artemisinin and 5-ALA combination therapy. Figure 3-2E: Image analysis and quantification of GFP-positive tumor areas in treated organoids. Eight organoids per group were analyzed on day 3 and normalized to day 1. Data are presented as box plots (25th-75th percentiles, median). Figure 3-2F: Flow cytometry analysis of dissociated brain tumor organoids. The relative percentage of GFP+ tumor cells in at least eight organoids on day 5, normalized to the control (DMSO), is shown. Data are presented as box plots (25th-75th percentiles, median). Figure 3-2G: ROS / DHE staining and flow cytometry analysis (PE, 582 / 15 nm) in dissociated tumor organoids. The mean fluorescence intensity (MFI) in DHE-stained GFP+ tumor cells and GFP- control cells is shown. n=2, values are mean ± SD. Figure 3-2H: Quantification of γH2AX+ cells in brain tumor organoids. Six cryosections from three organoids were stained with γH2AX, scanned, and 25 regions of interest (ROIs, 2,500 µm) were analyzed per condition. Raw values were normalized to organoid area (per ROI) and presented as box plots (25th–75th percentile, median). [Figure 4-1] Figure 4: Human glioblastoma-like tumor organoid model shows increased sensitivity to artemisinin and 5-ALA combination therapy. Figure 4-1A: Representative fluorescent images (left panel) and brightfield images (right panel) of human brain tumor organoids treated with 5-ALA and artemisinin. Organoids were monitored on days 1, 5, 8, and 11. Scale bar 500 µm. [Figure 4-2]Figure 4: Human glioblastoma-like tumor organoid model shows increased sensitivity to artemisinin and 5-ALA combination therapy. Figure 4-2B: Image analysis and quantification of treated organoids. The area of GFP-positive tumor cells on d11 was normalized to d1. Data are shown as box plots (25th-75th percentile, median) n=4. Figure 4-2C: Representative brightfield image of a patient-derived glioblastoma spheroid (VBT92) treated with 5-ALA and artemisinin. Images were taken on day 3 of culture and treatment. [Figure 5-1] Figure 5 (related to Figure 1): Artemisinin titration curve and overview of the porphyrin biosynthetic pathway. Figure 5-1A,B: Cell viability of (A) mouse ESCs and (B) primary (MEFp3) fibroblasts, as well as mouse (B16F10, 4T1) and human (MDA-MB-231, Mcf7, Panc1) cancer cells treated with artemisinin. Viability was assessed after 48 hours of treatment using Alamar Blue staining. [Figure 5-2] Figure 5 (related to Figure 1): Artemisinin titration curve and overview of the porphyrin biosynthetic pathway. Figure 5-2C: Porphyrin biosynthetic pathway component integration sites from artemisinin screening. Genomic locations of porphyrin biosynthetic genes (i.e., enzymes, in bold) and target introns and exons are shown, as well as retroviral (top) or Tol2 transposon (bottom) integration sites (vertical bars) in the forward (black) and reverse (gray) strands. [Figure 6-1]Figure 6 (related to Figure 2): Effects of combined artemisinin and 5-ALA treatment on cancer cell line viability, ROS production, and mitochondrial membrane potential. Figure 6-1A: Cell viability of artemisinin-treated mouse cancer cells (Mcf7-breast cancer; B16F10-melanoma) in the presence and absence of 5-ALA (0.5 mM). Alamar Blue was used to determine viability after 48 hours. Figure 6-1B,C: Cell viability of artemisinin-treated primary human glioblastoma cells in the presence or absence of 5-ALA. CellTiter-Glo was used to assess viability at 72 hours. Values are means ± SD. Figure 6-1D: ROS / DHE staining and flow cytometry analysis of Jurkat cells treated with piperlongumine or artemisinin (D) (48 hours). Values are means ± SD. [Figure 6-2] Figure 6 (related to Figure 2): Effects of combined artemisinin and 5-ALA treatment on cancer cell line viability, ROS production, and mitochondrial membrane potential. Figure 6-2E: ROS / DHE staining and flow cytometry analysis of (E) HL-60 cells treated with piperlongumine or artemisinin for 48 hours. Values are mean ± SD. Figure 6-2F, G, and H: (F) ROS levels (DHE staining, PE582 / 15nm-MFI), (G) cell viability, and (H) JC-1 levels in Jurkat cells treated with artemisinin (0.5 μM), 5-ALA (0.25 mM), or Ppox inhibitor (10 μM) for 48 hours. JC-1-negative cells have a decrease in mitochondrial membrane potential associated with apoptotic cell death. DHE fluorescence, relative cell number, and percentage of JC-1-negative cells were assessed using high-throughput flow cytometry and automated cell counting. All experiments were performed in triplicate and repeated once (JC-1) or twice (DHE, cell viability). Values are means ± SD. [Figure 7-1]Figure 7 (related to Figure 3): Cancer cell populations in CNS-PNET tumor organoids exhibit high sensitivity to artemisinin and 5-ALA combination therapy. Figure 7-1A: Cell viability in dissociated CNS-PNET tumor organoids. Quantification of GFP+ tumor cells and GFP- control cells in treated organoids, normalized to the control (DMSO), is shown. Data are presented as box plots (25th–75th percentiles, median). [Figure 7-2] Figure 7 (related to Figure 3): Cancer cell populations in CNS-PNET tumor organoids are highly sensitive to artemisinin and 5-ALA combination therapy. Figure 7-2B: Representative fluorescence (left panel) and brightfield images (right panel) of brain tumor organoids treated with control (DMSO), 5-ALA (0.0625 mM), artemisinin (1 μM), or 5-ALA + artemisinin (0.0625 mM and 1 μM). Scale bar 500 μM. [Figure 7-3] Figure 7 (related to Figure 3): Cancer cell populations in CNS-PNET tumor organoids exhibit high sensitivity to artemisinin and 5-ALA combination therapy. Figure 7-3C: Image quantification of GFP-positive tumor areas on day 5 of treatment compared to day 1. Box plots of data (25th–75th percentiles, median) are shown. [Figure 7-4] Figure 7 (related to Figure 3): Cancer cell populations in CNS-PNET tumor organoids are highly sensitive to artemisinin and 5-ALA combination therapy. Figure 7-4D: Representative images of anti-Sox2, anti-GFP, and DAPI stained sections in artemisinin and 5-ALA treated organoids. Scale bar 50 μm. [Figure 8] Figure 8 (related to Figure 3): Staining of CNS-PNET tumor organoid sections after artemisinin and 5-ALA treatment. Representative fluorescent (anti-GFP, DAPI) and H&E (hematoxylin & eosin staining) images of fixed frozen sections of control and treated tumor organoids. Areas of rosette-like structures (R) or tumor tissue (T) are indicated and magnified (6.8x). Scale bar 500 μM. [Figure 9-1]Figure 9 (related to Figure 3): Analysis of CNS-PNET tumor organoids after artemisinin and 5-ALA treatment. Figure 9-1A: Representative FACS plot of ROS / DHE-stained dissociated tumor organoids. Flow cytometry analysis (PE, 582 / 15 nm) of GFP+ tumor cells and GFP- wild-type cells is shown. Figure 9-2B: Representative microscopic image of a tumor organoid section stained for γH2AX, GFP, and DAPI. Scale bar 50 μM. [Figure 9-2] Figure 9 (related to Figure 3): Analysis of CNS-PNET tumor organoids after artemisinin and 5-ALA treatment. Figure 9-2C: Representative image and analysis mask of a γH2AX-stained and scanned tumor organoid slide. [Figure 10-1] Figure 10 (related to Figure 3): Serial analysis of CNS-PNET tumor organoids after artemisinin and 5-ALA treatment. Figure 10-1A,B: Quantification of (A) caspase 3 (Casp3) and (B) Ki67-positive cells in 5-ALA- and artemisinin-treated tumor organoids. For each condition and group, six sections of each of three organoids were stained with caspase 3 or Ki67, imaged using a high-magnification fluorescent scanner, and 25 regions of interest (ROIs, 2,500 µm) were selected and analyzed. The number of Casp3- or Ki67-positive cells was normalized to the analyzed area (per ROI, GFP+, or GFP-) and is shown as a box plot (median, 25th–75th percentile). [Figure 10-2] Figure 10 (related to Figure 3): Serial analysis of CNS-PNET tumor organoids after artemisinin and 5-ALA treatment. Figure 10-2C: Representative images of anti-Casp3, anti-GFP, and DAPI stained sections. Scale bar 50 μm. [Figure 11-1] Figure 11 (related to Figure 4): Combination therapy of artemisinin and 5-ALA in a human glioblastoma-like tumor organoid model. Figure 11-1A: Fluorescence images of 5-ALA and artemisinin-treated brain tumor organoids. Organoids were monitored on days 1, 5, 8, and 11. Scale bar 500 μm. [Figure 11-2] Figure 11 (related to Figure 4): Combination therapy of artemisinin and 5-ALA in a human glioblastoma-like tumor organoid model. Figure 11-2B: Image analysis and quantification of GFP-positive tumor areas in treated organoids. Organoids were analyzed on day 8 and normalized to day 1. Data are shown as box plots (25th–75th percentiles, median). [Figure 12-1] Figure 12: Artemisinin (ART) and its derivatives dihydroartemisinin (DHA) and artesunate (ARS), when combined with 5-aminolevulinic acid (5-ALA), exert synergistic antiproliferative effects on glioblastoma cell lines and mouse embryonic stem cells (ESCs). Shown are 6-day titration curves of artemisinin compounds and 5-ALA. Figures 12-1A-C: Cell viability of glioblastoma lines (as indicated) and ESCs treated with 5-ALA in combination with (A) ART administration, or (B) DHA administration, or (C) ARS administration, compared to untreated controls. Values are means ± SEM from three or more independent experiments. The dotted line indicates 100% survival. [Figure 12-2] Figure 12: Artemisinin (ART) and its derivatives dihydroartemisinin (DHA) and artesunate (ARS), when combined with 5-aminolevulinic acid (5-ALA), exert synergistic antiproliferative effects on glioblastoma cell lines and mouse embryonic stem cells (ESCs). Shown are 6-day titration curves of artemisinin compounds and 5-ALA. Figure 12-2, D-F: Cell viability of glioblastoma lines (as indicated) and ESCs treated with 5-ALA in combination with (D) ART administration, (E) DHA administration, or (F) ARS administration, compared to untreated controls. Values are means ± SEM from three or more independent experiments. The dotted line indicates 100% survival. [Figure 12-3]Figure 12: Artemisinin (ART) and its derivatives dihydroartemisinin (DHA) and artesunate (ARS), when combined with 5-aminolevulinic acid (5-ALA), exert synergistic antiproliferative effects on glioblastoma cell lines and mouse embryonic stem cells (ESCs). Shown are 6-day titration curves of artemisinin compounds and 5-ALA. Figures 12-3, G-I: Cell viability of glioblastoma lines (as indicated) and ESCs treated with 5-ALA in combination with (G) ART administration, (H) DHA administration, or (I) ARS administration, compared to untreated controls. Values are means ± SEM from three or more independent experiments. The dotted line indicates 100% survival. [Figure 12-4] Figure 12: Artemisinin (ART) and its derivatives dihydroartemisinin (DHA) and artesunate (ARS), when combined with 5-aminolevulinic acid (5-ALA), exert synergistic antiproliferative effects on glioblastoma cell lines and mouse embryonic stem cells (ESCs). Shown are 6-day titration curves of artemisinin compounds and 5-ALA. Figures 12-4 J-L: Cell viability of glioblastoma lines (as indicated) and ESCs treated with 5-ALA in combination with (J) ART administration, (K) DHA administration, or (L) ARS administration, compared to untreated controls. Values are means ± SEM from three or more independent experiments. The dotted line indicates 100% survival. [Figure 12-5] Figure 12: Artemisinin (ART) and its derivatives dihydroartemisinin (DHA) and artesunate (ARS), when combined with 5-aminolevulinic acid (5-ALA), exert synergistic antiproliferative effects on glioblastoma cell lines and mouse embryonic stem cells (ESCs). Shown are 6-day titration curves of artemisinin compounds and 5-ALA. Figure 12-5 MO: Cell viability of glioblastoma lines (as indicated) and ESCs treated with 5-ALA in combination with (M) ART administration, (N) DHA administration, or (O) ARS administration, compared to untreated controls. Values are means ± SEM from three or more independent experiments. The dotted line indicates 100% survival. [Figure 12-6]Figure 12: Artemisinin (ART) and its derivatives dihydroartemisinin (DHA) and artesunate (ARS), when combined with 5-aminolevulinic acid (5-ALA), exert synergistic antiproliferative effects on glioblastoma cell lines and mouse embryonic stem cells (ESCs). Shown are 6-day titration curves of artemisinin compounds and 5-ALA. Figure 12-6 P-R: Cell viability of glioblastoma lines (as indicated) and ESCs treated with 5-ALA in combination with (P) ART administration, (Q) DHA administration, or (R) ARS administration, compared to untreated controls. Values are means ± SEM from three or more independent experiments. The dotted line indicates 100% survival. [Figure 13-1] Figure 13: ART or DHA in combination with 5-ALA increases the antiproliferative effect of temozolomide (TMZ) treatment in glioblastoma cell lines and ESCs. Figures 13-1A-D: Glioblastoma lines (as indicated) and ESCs treated with 5-ALA and TMZ in combination with the indicated (A, C) ART administration, or (B, D) DHA administration, compared to untreated controls. Values are means + SEM from 5 or more independent experiments. Dotted lines indicate 100% survival and survival relative to TMZ treatment alone. [Figure 13-2] Figure 13: ART or DHA in combination with 5-ALA increases the antiproliferative effect of temozolomide (TMZ) treatment in glioblastoma cell lines and ESCs. Figures 13-2 E-H: Glioblastoma lines (as indicated) and ESCs treated with 5-ALA and TMZ in combination with the indicated (E, G) ART administration, or (F, H) DHA administration, compared to untreated controls. Values are means + SEM from ≥5 independent experiments. Dotted lines indicate 100% survival and survival relative to TMZ treatment alone. [Figure 13-3]Figure 13: ART or DHA in combination with 5-ALA increases the antiproliferative effect of temozolomide (TMZ) treatment in glioblastoma cell lines and ESCs. Figures 13-3 I-L: Glioblastoma lines (as indicated) and ESCs treated with 5-ALA and TMZ in combination with the indicated (I,K) ART administration, or (J,L) DHA administration, compared to untreated controls. Values are means + SEM from 5 or more independent experiments. Dotted lines indicate 100% survival and survival relative to TMZ treatment alone. [Figure 14-1] Figure 14: ART in combination with 5-ALA or methyl-5-ALA increases the antiproliferative effect of temozolomide (TMZ) treatment in glioblastoma cell lines and ESCs. 5-ALA is more potent than methyl-5-ALA in this setup. Figure 14-1A,B: Glioblastoma lines (as indicated) and ESCs treated with 5-ALA and TMZ in combination with the indicated ART doses compared to untreated controls. Values are means + SEM from three or more independent experiments. Dotted lines indicate 100% survival and survival relative to TMZ treatment alone. [Figure 14-2] Figure 14: ART in combination with 5-ALA or methyl-5-ALA increases the antiproliferative effect of temozolomide (TMZ) treatment in glioblastoma cell lines and ESCs. 5-ALA is more potent than methyl-5-ALA in this setup. Figure 14-2C-F: Glioblastoma lines (as indicated) and ESCs treated with 5-ALA and TMZ in combination with the indicated ART doses compared to untreated controls. Values are means + SEM from three or more independent experiments. Dotted lines indicate 100% survival and survival relative to TMZ treatment alone. [Figure 15] Figure 15: ART in combination with 5-ALA increases the antiproliferative effect of lomustine (CCNU) treatment in glioblastoma cell lines and ESCs. Figures 15A-F: Glioblastoma lines (as indicated) and ESCs treated with 5-ALA and CCNU in combination with ART administration compared to untreated controls. Values are means + SEM from at least four independent experiments. Dotted lines indicate 100% survival and survival with CCNU treatment alone. [Figure 16-1] Figure 16: ART, DHA, or ARS in combination with 5-ALA increases the antiproliferative effects of cisplatin (CP) and 5-fluorouracil (5-FU) in two lung cancer cell lines. Figures 16-1A-F: Cell survival of 5-ALA-treated lung cancer lines (as indicated) in combination with the indicated (A, D) ART, (B, E) DHA, or (C, F) ARS treatment, and with cisplatin (A-F), compared to untreated controls. Values are means + SEM from four or more independent experiments. Dotted lines indicate 100% survival and survival relative to chemotherapy (cisplatin or 5-FU) treatment alone. [Figure 16-2] Figure 16: ART, DHA, or ARS in combination with 5-ALA increases the antiproliferative effects of cisplatin (CP) and 5-fluorouracil (5-FU) in two lung cancer cell lines. Figures 16-2, G-L: Cell survival of 5-ALA-treated lung cancer lines (as indicated) in combination with the indicated (G,J) ART, (H,K) DHA, or (I,L) ARS treatment, and 5-FU (G-L), compared to untreated controls. Values are means + SEM from at least four independent experiments. Dotted lines indicate 100% survival and survival relative to chemotherapy (cisplatin or 5-FU) treatment alone. [Figure 17] Figure 17: ART, DHA, or ARS in combination with 5-ALA enhances the antiproliferative effects of cisplatin (CP) and 5-fluorouracil (5-FU) in HepG2 cells (liver cancer). Figures 17A-F: Cell survival of 5-ALA-treated HepG2 cells compared to untreated controls in combination with the indicated (A, D) ART, (B, E) DHA, or (C, F) ARS treatments, and either cisplatin (A-C) or 5-FU (D-F). Values are means + SEM from four or more independent experiments. Dotted lines indicate 100% survival and survival relative to chemotherapy (cisplatin or 5-FU) treatment alone. [Figure 18]Figure 18: ART, DHA, or ARS in combination with 5-ALA enhances the antiproliferative effects of cisplatin and 5-fluorouracil (5-FU) in MD-MBA-231 cells (breast cancer cells). Figures 18A-F: Cell survival of 5-ALA-treated MD-MBA-231 cells compared to untreated controls in combination with the indicated (A, D) ART, (B, E) DHA, or (C, F) ARS treatments, and either cisplatin (A-C) or 5-FU (D-F). Values are means + SEM from four or more independent experiments. Dotted lines indicate 100% survival and survival relative to chemotherapy (cisplatin or 5-FU) treatment alone. [Figure 19] Figure 19: ART, DHA, or ARS in combination with 5-ALA enhances the antiproliferative effects of cisplatin and 5-fluorouracil (5-FU) in MiaPaca-2 cells (pancreatic cancer). Figures 19A-F: Cell survival of MiaPaca-2 cells treated with 5-ALA in combination with the indicated (A,D) ART, (B,E) DHA, or (C,F) ARS, and with either cisplatin (A-C) or 5-FU (D-F), compared to untreated controls. Values are means + SEM from four or more independent experiments. Dotted lines indicate 100% survival and survival relative to chemotherapy (cisplatin or 5-FU) treatment alone. DETAILED DESCRIPTION OF THE INVENTION

[0189] [Example] [material and method] [Mammalian tissue culture] Mouse embryonic stem cell clones (clone AN3-12) {Elling:2011gla} were cultured in DMEM supplemented with 10% fetal calf serum (FCS), penicillin-streptomycin, non-essential amino acids, sodium pyruvate (1 mM), l-glutamine (2 mM), β-mercaptoethanol (0.1 mM), and LIF (20 μg ml-1). SH-SY5Y cells were cultured in DMEM / F12 1:1 supplemented with 10% fetal calf serum (FCS), penicillin-streptomycin, and l-glutamine. 4T1 cells were cultured in IMDM supplemented with 10% fetal calf serum (FCS), penicillin-streptomycin, and l-glutamine. MEFs, Mcf7, MDA-MB-231, Panc1, LN229, A549, MiaPaca-2, B16F10, and PlatE cells were cultured in DMEM supplemented with 10% FCS, penicillin-streptomycin, and L-glutamine. HepG2, T98G, and U87MG cells were cultured in EMEM, and SHP77, VBT92, and VBT281 cells were cultured in RPMI, each supplemented with 10% FCS, penicillin-streptomycin, and L-glutamine. All cells were cultured at 37°C in a 20% O2 and 5% CO2 atmosphere.

[0190] [Cell line] The mouse AN3-12 ESC line was generated in our laboratory and characterized and authenticated as previously described {Elling, U. et al. Forward and reverse genetics through derivation of haploid mouse embryonic stem cells. Cell stem Cell 9, 563-574 (2011)}. Haploid mouse ESCs were used for the derivation of insertional mutagenesis and gene trap knockout cell lines. SH-SY5Y cells were obtained directly from the supplier (Sigma-Aldrich) and used for proliferation assays and cell staining. Jurkat cells used in the in vitro viability and DHE assays were obtained from an in-house source and have been functionally described elsewhere. {Reikerstorfer, A., Holz, H., Stunnenberg, H.G., & Busslinger, M. Low affinity binding of interleukin-1 beta and intracellular signaling via NF-kappa B identify Fit-1 as a distant member of the interleukin-1 receptor family. The Journal of Biological Chemistry 270, 17645-17648 (1995)}. Mcf7, MDA-MB-231, 4T1, Panc1, and B16F10 cancer cell lines were obtained in-house. MEFs were generated and obtained in our laboratory.PlatE cells were used for recombinant retrovirus and lentivirus production as previously described (Taubenschmid, J. et al., A vital sugar code for ricin toxicity. Cell Research 27, 1351-1364 (2017). Stadlmann, J. et al., Comparative glycoproteomics of stem cells identifies new players in ricin toxicity. Nature 549, 538-542 (2017)). All cell lines tested negative for mycoplasma. Cell lines listed by ICLAC were not used.

[0191] [Competitive growth analysis] Haploid ESCs carrying a gene trap in the nomic intron were seeded at low density in standard ESC growth medium and infected with two viruses, one encoding mCherry and Cre recombinase and the other encoding GFP, both with puromycin (Invivogen, ant-pr-1), for 12 hours. Infected cells were selected (final concentration of puromycin, 1 μg / ml) and expanded 24 hours later. The ratio of GFP-expressing cells to mCherry / Cre-expressing cells in the presence and absence of artemisinin was determined using high-throughput flow cytometry (BD LSR Fortessa HTS cell analyzer).

[0192] [Chemical profiling in cell lines] For dose response, cells were seeded in 96-well plates (25,000 cells / 96-well plate in triplicate, where indicated) and exposed to compounds for 48 hours. Cell viability was assessed using automated cell counting (high-throughput flow cytometry), Alamar Blue staining (Invitrogen, DAL1100), or CellTiter-Glo luminescence assay (Promega, G7570, according to the manufacturer's protocol), respectively. Cell viability assays (Figures 12-19) were performed in technical duplicates in 96-well plates. Treatments were initiated 24 hours after cell plating and continued for 6 days, and cell viability was assessed using a Cell Titer Glo 2.0 luminescence assay (Promega, G9242).

[0193] [Dihydroethidium (DHE) staining] Treated cells were harvested, washed with 1x HBSS (Ca2+- and Mg2+-free), incubated with 1 mM DHE (dihydroethidium (hydroethidine), Invitrogen, D11347) in 1x HBSS for 45 min at 37°C, washed twice, and counterstained with DAPI or viability dye (eBioscience™ Fixable Viability Dye eFluor™ 780, 65-0865-18) for 10 min on ice. Cells were then collected, strained, and analyzed for DHE using flow cytometry in the red fluorescence spectrum (PE channel).

[0194] [JC-1 staining] The cells were harvested, washed with 1x PBS, incubated with a solution of 2 μM JC-1 in 1 PBS (MitoProbe JC-1 Assay Kit-1, Invitrogen, M34152) for 35 min at 37°C, washed twice, and analyzed by flow cytometry in red (PE channel) and green (FITC channel) fluorescence spectra.

[0195] [Cerebral organoid formation] Cerebral organoids were generated as previously described (Lancaster, MA et al. Cerebral organoids model human brain development and microcephaly. Nature 501, 373-379 (2013)). Human embryonic stem cells (feeder-free H9, WiCell) were transferred to low-attachment 96-well plates (Corning) at a density of 9,000 cells per well and incubated in human stem cell medium. After 6 days, the medium was changed to neural induction medium containing Dulbecco's Modified Eagle's Medium (DMEM / F12), N2 supplement (Invitrogen), Glutamax (Invitrogen), minimal essential medium with non-essential amino acids (MEM-NEAA), and 1 μg / ml heparin (Sigma) to promote the growth of ectodermal tissue. On day 11, embryoid bodies (EBs) were embedded in droplets of Matrigel and transferred to differentiation medium containing DMEM / F12:Neurobasal 1:1, N2 supplement (Invitrogen), B27 supplement (without vitamin A) (Invitrogen), 50 μM 2-mercaptoethanol, 1:4,000 insulin (Sigma), Glutamax (Invitrogen), penicillin-streptomycin, and MEM-NEAA on 10 cm plates. After 5 days, organoids were transferred to an orbital shaker and maintained in differentiation medium (B27 supplement) with vitamin A.

[0196] [Brain tumor organoid formation + nucleofection] Tumor initiation was induced in day 10 embryoid bodies (EBs) by either oncogene amplification using Sleeping Beauty (SB) transposase or tumor suppressor gene mutation using the CRISPR-Cas9 system. Plasmids expressing the transposase, GFP, and the desired oncogene or Cas9 nuclease were introduced by electroporation. Briefly, a mixture of 1 μg of DNA and 100 μl of nucleofector solution was added to 10 EBs and transferred to a nucleofection cuvette. Lonza Nucleofector 2b and the A-023 program were used for electroporation / nucleofection. EBs were then transferred to a 10 cm dish containing differentiation medium containing vitamin A and embedded in Matrigel 24 hours later. Two distinct tumor types were generated {Bian:2018gs}: central nervous system primitive neuroectodermal tumor (CNS-PNET) was induced by overexpression of Myc. Glioblastoma-like tumor group 2 (GBM-2) was caused by mutation of the tumor suppressor genes p53, NF1, and PTEN. All plasmids were designed by Shan Bian {Bian, S. et al. Genetically engineered cerebral organoids model brain tumor formation. Nat. Methods 15, 631-639 (2018)}.

[0197] [Chemical profiling in brain tumor organoids] Brain tumor organoids were treated with various compounds, and the survival and growth of transformed and non-transformed neurons were monitored. Fluorescent labeling of tumor cells allowed clear differentiation of transformed cells compared to unlabeled cells throughout the experiment using brightfield microscopy and fluorescence imaging. Transformed tumor tissue and non-transformed neurons within the same organoid were monitored throughout the experiment using fluorescence imaging and brightfield microscopy. At the end of treatment (day 5 or day 7, respectively), the number of GFP-positive cells was assessed by flow cytometry.

[0198] [Flow cytometry analysis of brain tumor organoids] Brain (tumor) organoids were enzymatically and mechanically dissociated using 1x trypsin, incubated at 37°C for 35-45 minutes, and gently shaken. Organoids were singularized by carefully resuspending, adding differentiation medium, and straining (Falcon round-bottom tubes with cell strainer caps, 5 mL, 35 μm nylon mesh cell strainer snap caps). Single cells in suspension were counterstained with DAPI or a viability dye (eBioscience™ Fixable Viability Dye eFluor™ 780, 65-0865-18) and analyzed using flow cytometry (BD LSR Fortessa HTS cell analyzer). The amount of GFP-positive cells was analyzed.

[0199] [Imaging of brain tumor organoids] 5-ALA- or artemisinin-treated brain organoids were imaged at the indicated time points using an Axio Vert. A1 inverted microscope system (Zeiss EC Plan-Neofluar 2.5x / 0.085 Pol M27 objective, 0.5 camera adapter). Bright-field and green fluorescent images were taken from the same area. Further image analysis was performed using Celldiscoverer 7 (Zeiss), a fully integrated, high-performance automated live-cell imaging system.

[0200] [Immunohistochemical staining] Brain organoids were fixed in 4% PFA (room temperature, 1 h), incubated with 30% sucrose (o / n, 4°C), embedded in OCT (Tissue-tek OCT Compound, SANOVA PHARMA GESMBH, 4583), and cryostat-cut into 20 μm sections (-12 / -14°C). For staining, sections were blocked and permeabilized in 0.25% Triton X-100 and 4% donkey serum in PBS for 1 hour at room temperature, stained overnight with primary antibodies diluted in 0.1% Triton X-100 and 4% donkey serum in PBS, incubated with secondary antibodies diluted in 0.1% Triton X-100 and 4% donkey serum in PBS for 2 hours at room temperature, and counterstained with DAPI (4',6-diamidino-2-phenylindole, dilactate, Invitrogen, D3571) for 20 minutes at room temperature. Fluorescent mounting medium (Dako, S302380) was used to mount the specimen slides. Organoids were imaged on an LSM780 Axio Imager (point laser scanning confocal microscope, GaAsP (gallium arsenide) detector with 45% QE and up to 2× SNR) equipped with a standard filter set (CH1:371-735, CH2:479-735, CH3:Quasar (GaAsP)) through a 20× / 0.8 Plan-Apochromat objective (Carl Zeiss) using laser illumination (diode laser 405-25mW, Argon 458, 488, 514-30mW, DPSS 561-15mW, HeNe 633-5mW).

[0201] Statistics and Reproducibility Unless otherwise stated, all values in Figures 1-11 are given as mean ± SD. All experiments were independently replicated two to seven times with similar results. Figures were prepared and statistical analyses were performed using GraphPad Prism (GraphPad Software). No a priori sample size estimation was performed. The experiment was not randomized. Investigators were not blinded to allocation during the experiment and outcome assessment. Data were analyzed by using an unpaired, two-tailed Student's t-test as indicated. P<0.05 was accepted as statistically significant. Box plots represent the median and range from the first to third quartile.

[0202] For the data shown in Figures 12-19, all values are given +SEM. Data were analyzed by using a paired two-tailed Student's t-test according to the experimental design. Significance is indicated as follows: * p<0.05, ** p<0.01, *** p<0.001.

Claims

1. a) 【Chemical 1】 An artemisinin compound (1) selected from: b) 5-aminolevulinic acid (2), methyl-5-aminolevulinic acid (2b), or a pharmaceutically acceptable salt or solvate thereof; and c) at least one chemotherapeutic agent selected from temozolomide and lomustine; A pharmaceutical composition comprising:

2. 2. The pharmaceutical composition according to claim 1, wherein the molar ratio of the artemisinin compound (1) to the 5-aminolevulinic acid (2) or the methyl-5-aminolevulinic acid (2b) is in the range of 1:5 to 1:5000.

3. 3. The pharmaceutical composition of claim 1 or claim 2, further comprising a pharmaceutically acceptable carrier, excipient and / or diluent.

4. The pharmaceutical composition according to any one of claims 1 to 3, for use in the prevention and / or treatment of cancer selected from hematopoietic cancer, brain cancer, pancreatic cancer, liver cancer, breast cancer and lung cancer.

5. 5. The pharmaceutical composition of claim 4, wherein the pharmaceutical composition is in the form of a tablet, capsule, syrup, solution, suspension, emulsion, or gel.

6. The pharmaceutical composition according to any one of claims 1 to 5, wherein the pharmaceutical composition is administered by oral, intrathecal, intravenous, subcutaneous, parenteral application or by inhalation.

7. The pharmaceutical composition according to any one of claims 1 to 6, wherein the artemisinin compound (1) is administered in a range of 0.01 to 100 mg / kg of body weight per day, and the 5-aminolevulinic acid or the methyl-5-aminolevulinic acid is administered in a range of 0.01 to 200 mg / kg of body weight per day.

8. The pharmaceutical composition according to any one of claims 1 to 7, wherein the at least one chemotherapeutic agent is administered in the range of 0.01 to 100 mg / kg of body weight per day.

9. 5. The pharmaceutical composition of claim 4, wherein the brain cancer is selected from proneural (PN), mesenchymal (MES), and classical (CL) glioblastoma subtypes.

10. 5. The pharmaceutical composition of claim 4, wherein the prevention and / or treatment of cancer selected from hematopoietic cancer, brain cancer, pancreatic cancer, liver cancer, breast cancer and lung cancer is performed in combination with radiation therapy, immunotherapy, electromagnetic field therapy, hyperthermia, chemotherapy, cancer immunotherapy, and / or any other small molecule-based therapy.

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