Patent Application
20230381176
The present invention pertains to the biomedical field, and particularly relates to a method for treating or suppressing a cancer, reducing its severity, lowering its risk or inhibiting its metastasis in an individual. The method comprises administering to the individual a therapeutically effective amount of an ALK inhibitor, and a therapeutically effective amount of one or more other anticancer reagents. The invention also relates to a pharmaceutical composition or kit comprising an ALK inhibitor, and one or more other anticancer reagents.
Proliferative diseases represent a serious threat to modern society. Cancerous growths pose serious challenges for modern medicine due to their unique characteristics, including uncontrollable cell proliferation, an ability to invade local and even remote tissues, lack of differentiation, lack of detectable symptoms and lack of effective therapy and prevention. Worldwide, more than 10 million people are diagnosed with cancer every year, and cancer causes six million deaths every year or 12% of the deaths worldwide.
Small molecule ALK inhibitors can be designed to target ALK mutations, so as to treat related diseases and conditions including cancer. ALK mutations targeted drugs include: crizotinib as first-generation, ceritinib, alectinib and brigatinib as second-generation, lorlatinib as third-generation, and the like. However, the targeted drugs usually exhibit resistance about 1 year after administration. Thus, overcoming the drug resistance of the targeted drugs as well as other anticancer reagents and improving the efficacy are some of the main objectives in drug research and development.
In one aspect, the invention provides a method for treating or suppressing a cancer, reducing its severity, lowering its risk or inhibiting its metastasis in an individual, said method comprising administering to the individual a therapeutically effective amount of an ALK inhibitor, and a therapeutically effective amount of one or more anticancer reagents selected from a CDK4/6 inhibitor, an Mek inhibitor, a BRAF inhibitor, a chemotherapeutic agent, an MDM2 inhibitor, an HDAC inhibitor, a PD-1 inhibitor, a PARP inhibitor, a VEGF inhibitor and a BCR-ABL inhibitor.
In another aspect, the invention provides a use of an ALK inhibitor in manufacture of a medicament in combination with one or more anticancer reagents selected from a CDK4/6 inhibitor, an Mek inhibitor, a BRAF inhibitor, a chemotherapeutic agent, an MDM2 inhibitor, an HDAC inhibitor, a PD-1 inhibitor, a PARP inhibitor, a VEGF inhibitor and a BCR-ABL inhibitor for treating or suppressing a cancer, reducing its severity, lowering its risk or inhibiting its metastasis in an individual.
In another aspect, the invention provides a use of a pharmaceutical composition for treating or suppressing a cancer, reducing its severity, lowering its risk or inhibiting its metastasis in an individual, said pharmaceutical composition comprising an ALK inhibitor as well as one or more anticancer reagents selected from a CDK4/6 inhibitor, an Mek inhibitor, a BRAF inhibitor, a chemotherapeutic agent, an MDM2 inhibitor, an HDAC inhibitor, a PD-1 inhibitor, a PARP inhibitor, a VEGF inhibitor and a BCR-ABL inhibitor.
In another aspect, the invention provides an ALK inhibitor which is used in combination with one or more anticancer reagents selected from a CDK4/6 inhibitor, an Mek inhibitor, a BRAF inhibitor, a chemotherapeutic agent, an MDM2 inhibitor, an HDAC inhibitor, a PD-1 inhibitor, a PARP inhibitor, a VEGF inhibitor and a BCR-ABL inhibitor for treating or suppressing a cancer, reducing its severity, lowering its risk or inhibiting its metastasis in an individual.
In another aspect, the invention provides a pharmaceutical composition comprising an ALK inhibitor, and one or more anticancer reagents selected from a CDK4/6 inhibitor, an Mek inhibitor, a BRAF inhibitor, a chemotherapeutic agent, an MDM2 inhibitor, an HDAC inhibitor, a PD-1 inhibitor, a PARP inhibitor, a VEGF inhibitor and a BCR-ABL inhibitor, as well as a pharmaceutically acceptable carrier.
In another aspect, the invention provides a kit, comprising:
Unless otherwise defined below, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art. References to techniques used herein are intended to refer to techniques that are generally understood in the art, including those obvious changes or equivalent replacements of the techniques for those skilled in the art. While it is believed that the following terms are well understood by those skilled in the art, the following definitions are set forth to better explain the invention.
As used herein, the terms “including”, “comprising”, “having”, “containing” or “comprising”, and other variants thereof, are inclusive or open, and do not exclude other unlisted elements or method steps.
As used herein, “ALK” refers to anaplastic lymphoma kinase, and “ALK inhibitor” refers to an agent having an inhibitory effect on ALK. In some embodiments, the ALK inhibitor also has an inhibitory effect on one or more other targets (e.g., FAK (focal adhesion kinase) and/or ROS1 (a tyrosine protein kinase encoded by ROS1 proto-oncogene in human)).
As used herein, “CDK4/6 inhibitor” refers to an agent that selectively and efficiently inhibits cyclin-dependent kinase 4 or cyclin-dependent kinase 6 (CDK4/6).
As used herein, “Mek inhibitor” refers to an agent that inhibits mitogen-activated protein kinase (MEK), and MEK is a major protein in the RAS/RAF/MEK pathway, which signals toward cell proliferation and survival, and frequently activated in tumors that have mutations in the RAS or RAF oncogenes or in growth receptor tyrosine kinases.
As used herein, BRAF inhibitor refers to an agent that inhibits BRAF, such as Dabrafenib, Sorafenib, Regorafenib, Pazopanib, Vemurafenib etc.
As used herein, “chemotherapeutic agent” refers to chemotherapeutic drugs that can kill tumor cells, and these drugs can act on different stages of tumor cell growth and reproduction, thereby inhibit or kill tumor cells.
As used herein, “MDM2 inhibitor” refers to Murine Double Minute 2 (MDM2) inhibitor which interferes with the binding of MDM2 oncoprotein to the tumor suppressor p53 protein, and serves as pharmacological p53 activators.
As used herein, “HDAC inhibitor” refers to an agent that inhibits histone deacetylases (HDAC), and has been described to cause growth arrest with subsequent differentiation or apoptosis of tumor cells, whereas normal cells are not affected.
As used herein, “PD-1 inhibitor” refers to an agent that targets the programmed death 1 (PD-1) signaling pathway, and can be anti-PD-1 antibody. The anti-PD-1 antibody can be monoclonal antibody or bispecific antibody, it may be full length antibody or antibody fragment, as long as it can block the binding between PD-1 and PD-L1.
As used herein, “PARP inhibitor” refers to an agent that inhibits poly ADP ribose polymerase (PARP).
As used herein, “VEGF inhibitor” refers to an agent that targets vascular endothelial growth factor (VEGF) signaling pathway, wherein VEGF is a major regulatory factor of angiogenesis, and in most human tumors is involved in tumor growth and metastasis.
As used herein, “BCR-ABL inhibitor” refers to an agent that targets the fusion gene of abelson murine leukemia (Abl) and breakpoint cluster region (Bcr).
The term “alkyl” as used herein, alone or as part of another group, refers to an unsubstituted straight or branched aliphatic hydrocarbon containing from 1 to 12 carbon atoms (ie, C1-12 alkyl) or an indicated number of carbon atoms, for example, C1 alkyl such as methyl, C2 alkyl such as ethyl, C3 alkyl such as n-propyl or isopropyl, C1-3 alkyl such as methyl, ethyl, n-propyl or isopropyl, or the like. In one embodiment, the alkyl is C1-4 alkyl. Non-limiting examples of C1-12 alkyl include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, isobutyl, 3-pentyl, hexyl, heptyl, octyl, nonyl and decyl. Examples of C1-4 alkyl include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, and isobutyl.
The term “cycloalkyl” as used herein, alone or as part of another group, refers to a saturated or partially unsaturated (containing one or two double bonds) cyclic aliphatic hydrocarbon, which comprises 1 or 2 rings having 3 to 12 carbon atoms or an indicated number of carbon atoms (i.e., C3-12 cycloalkyl).
In one embodiment, the cycloalkyl has two rings. In one embodiment, the cycloalkyl has one ring. In another embodiment, the cycloalkyl group is selected from the group consisting of C3-8 cycloalkyl groups. In another embodiment, the cycloalkyl group is selected from the group consisting of C3-6 cycloalkyl groups. Non-limiting examples of cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, norbornyl, decahydronaphthyl, adamantyl, cyclohexenyl, and cyclopentenyl.
The term “heterocycle” or “heterocyclyl” as used herein, alone or as part of another group, refers to a saturated or partially unsaturated (e.g., comprising one or two double bonds) cyclic group, which comprises 1, 2 or 3 rings having 3 to 14 ring members (i.e., 3- to 14-membered heterocyclyl), wherein at least one carbon atom of one of the rings is replaced by a heteroatom. Each heteroatom is independently selected from the group consisting of atoms of oxygen, sulfur (including sulfoxide and sulfone) and/or nitrogen (which may be oxidized or quaternized). The term “heterocyclyl” is intended to include a group wherein —CH2— in the ring is replaced by —C(═O)—, for example, cyclic ureido (such as 2-imidazolidinone) and cyclic amido (such as β-lactam, γ-lactam, δ-lactam, ε-lactam) and piperazin-2-one. In one embodiment, the heterocyclyl is a 3- to 8-membered cyclic group comprising 1 ring and 1 or 2 oxygen and/or nitrogen atoms. In one embodiment, the heterocyclyl is a 4-, 5- or 6-membered cyclic group comprising 1 ring and 1 or 2 oxygen and/or nitrogen atoms. In one embodiment, the heterocyclyl is a 4- or 6-membered cyclic group comprising 1 ring and 1 or 2 oxygen and/or nitrogen atoms. The heterocyclyl can be attached to the remainder of molecule via any available carbon or nitrogen atom. Non-limiting examples of the heterocyclyl include dioxanyl, tetrahydropyranyl, 2-oxopyrrolidin-3-yl, piperazin-2-one, piperazin-2,6-dione, 2-imidazolidinone, piperidinyl, morpholinyl, piperazinyl, pyrrolidinyl and dihydroindolyl.
As used herein, the term “enantiomeric excess” or “ee” refers to a measure of how much one enantiomer is present relative to another enantiomer. For a mixture of R and S enantiomers, the enantiomeric excess in form of percentage is defined as |R−S|*100, wherein R and S respectively represents mole or weight parts thereof in the mixture, and R+S=1. After knowing the optical rotation of chiral substance, the enantiomeric excess in form of percentage is defined as ([α]obs/[α]max)*100, wherein [α]obs represents the optical rotation of the mixture of enantiomers, [α]max represents the optical rotation of pure enantiomer. Enantiomeric excess can be determined using a variety of analytical techniques, including NMR spectroscopy, chiral column chromatography, or optical rotation. The compound of the present invention may have an ee of about 70% or more, such as about 80% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more.
The term “pharmaceutically acceptable salt”, as used herein, includes both acid addition salts and base addition salts of a compound.
Suitable acid addition salts are formed from acids which form non-toxic salts. Examples include acetate, adipate, aspartate, benzoate, besylate, bicarbonate/carbonate, bisulfate/sulfate, borate, camphorsulfonate, citrate, cyclohexylaminosulfonate, ethanedisulfonate, ethanesulfonate, formate, fumarate, glucoheptonate, gluconate, glucuronate, hexafluorophosphate, hibenzate, hydrochloride/chloride, hydrobromide/bromide, hydroiodide/iodide, isethionate, lactate, malate, maleate, malonate, methanesulfonate, methylsulfate, naphthylate, 2-naphthalenesulfonate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate/hydrogen phosphate/dihydrogen phosphate, pyroglutamate, aldarate, stearate, succinate, tannate, tartrate, tosylate, trifluoroacetate and xinofoate.
Suitable base addition salts are formed from bases which form non-toxic salts. Examples include aluminum salts, arginine salts, benzathine benzylpenicillin salts, calcium salts, choline salts, diethylamine salts, diethanolamine salts, glycine salts, lysine salts, magnesium salts, meglumine salts, ethanolamine salts, potassium salts, sodium salts, tromethamine salts and zinc salts.
For a review of suitable salts, see “Handbook of Pharmaceutical Salts: Properties, Selection, and Use” by Stahl and Wermuth (Wiley-VCH, 2002). Methods for preparing the pharmaceutically acceptable salts of the compounds of the invention are known to those skilled in the art.
The term “solvate” as used herein is a substance formed by combination, physical binding and/or solvation of a compound of the invention with a solvent molecule, such as a disolvate, a monosolvate or a hemisolvate, wherein the ratio of the solvent molecule to the compound of the invention is about 2:1, about 1:1 or about 1:2, respectively. This kind of physical bonding involves ionization and covalent bonding (including hydrogen bonding) in different degrees. In some cases (e.g., when one or more solvent molecules are incorporated into crystal lattice of crystalline solid), the solvate can be isolated.
Thus, the solvate comprises both solution phase and isolatable solvates. The compounds of the invention may be in solvated forms with pharmaceutically acceptable solvents (such as water, methanol and ethanol), and the present application is intended to encompass both solvated and unsolvated forms of the compounds of the invention.
One type of solvate is a hydrate. “Hydrate” relates to a specific subset of solvates wherein the solvent molecule is water. Solvates generally function in the form of pharmacological equivalents. The preparation of solvates is known in the art, see for example, M. Caira et al, J. Pharmaceut. Sci., 93(3): 601-611 (2004), which describes the preparation of a solvate of fluconazole with ethyl acetate and water. Similar methods for the preparation of solvates, hemisolvates, hydrates and the like are described by van Tonder et al, AAPS Pharm. Sci. Tech., 5(1): Article 12 (2004) and A. L. Bingham et al, Chem. Commun. 603-604 (2001). A representative and non-limiting method for the preparation of solvate involves dissolving a compound of the invention in a desired solvent (organic solvent, water or a mixture thereof) at a temperature above 20° C. to about 25° C., and then the solution is cooled at a rate sufficient to form a crystal, and the crystal is separated by a known method such as filtration. Analytical techniques such as infrared spectroscopy can be used to confirm the presence of the solvent in the crystal of the solvate.
“Pharmaceutically acceptable carrier” in the context of the present invention refers to a diluent, adjuvant, excipient or vehicle together with which the therapeutic agent is administered, and which is suitable for contacting a tissue of human and/or other animals within the scope of reasonable medical judgment, and without excessive toxicity, irritation, allergic reactions, or other problems or complications corresponding to a reasonable benefit/risk ratio.
The pharmaceutically acceptable carriers that can be used in the pharmaceutical compositions or kits of the invention include, but are not limited to, sterile liquids such as water and oils, including those oils derived from petroleum, animals, vegetables or synthetic origins, for example, peanut oil, soybean oil, mineral oil, sesame oil, etc. Water is an exemplary carrier when the pharmaceutical composition is administered intravenously. It is also possible to use physiological saline and an aqueous solution of glucose and glycerin as a liquid carrier, particularly for injection. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, maltose, chalk, silica gel, sodium stearate, glyceryl monostearate, talc, sodium chloride, skimmed milk powder, glycerin, propylene glycol, water, ethanol and the like. The pharmaceutical composition may further contain a small amount of a wetting agent, an emulsifier or a pH buffering agent as needed. Oral formulations may contain standard carriers such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, magnesium carbonate, and the like. Examples of suitable pharmaceutically acceptable carriers are as described in Remington's Pharmaceutical Sciences (1990).
The pharmaceutical compositions and the components of the kit of the invention may act systemically and/or locally. For this purpose, they may be administered via a suitable route, for example by injection (e.g., intravenous, intraarterial, subcutaneous, intraperitoneal, intramuscular administration, including instillation) or transdermal administration; or by oral, buccal, nasal, transmucosal, topical administration, in form of ophthalmic preparation or by inhalation.
For these routes of administration, the pharmaceutical compositions and the components of the kit of the invention may be administered in a suitable dosage form.
The dosage forms include, but are not limited to, tablets, capsules, troches, hard candy, pulvis, sprays, creams, ointments, suppositories, gels, pastes, lotions, ointments, aqueous suspensions, injectable solutions, elixirs, syrups.
The term “container” as used herein refers to a container for holding a pharmaceutical component. This container can be used for preparation, storage, transportation and/or stand-alone/bulk sale, which is intended to include bottles, cans, vials, flasks, syringes, tubes (e.g., those used in cream products), or any other containers for preparation, containment, storage or distribution of a drug product.
The term “specification/instruction” as used herein refers to an insert, a tag, a label, etc., which records information about a pharmaceutical component located in the container. The information as recorded is typically determined by the regulatory agency (e.g., the United States Food and Drug Administration) that governs the area in which the product is to be sold. Preferably, the package leaflet specifically lists an indication for which the use of the pharmaceutical component is approved. The package leaflet can be made of any material from which information contained therein or thereon can be read. Preferably, the package leaflet is a printable material (e.g., paper, plastic, cardboard, foil, adhesive paper or plastic, etc.) on which the desired information can be formed (e.g., printed or applied).
The term “effective amount” as used herein refers to an amount of active ingredient that, after administration, will relieve to some extent one or more symptoms of the condition being treated.
As used herein, “individual” includes a human or a non-human animal Exemplary human individual includes a human individual (referred to as a patient) suffering from a disease (such as the disease described herein) or a normal individual. “Non-human animal” in the present invention includes all vertebrates, such as non-mammals (e.g., birds, amphibians, reptiles) and mammals, such as non-human primates, domestic animals, and/or domesticated animals (e.g., sheep, dogs, cats, cows, pigs, etc.).
As used herein, “cancer metastasis” refers to a cancer that spreads (metastasizes) from its original site to another area of the body. Almost all cancers have the potential to metastasize. Whether metastasis will occur depends on complex interactions between multiple tumor cell factors (including type of cancer, degree of maturation (differentiation) of tumor cells, location and age of cancer, and other factors that are not fully understood). There are three ways of metastasis: local expansion from a tumor to a surrounding tissue, arrival through bloodstream to a distant site, or arrival through lymphatic system to an adjacent or distant lymph node. Each cancer can have a representative diffusion route. Tumors are named according to their primary sites (for example, breast cancer that has metastasized to the brain is called metastatic breast cancer that metastasizes to the brain).
As used herein, “resistance” refers to that a cancer cell is resistant to chemotherapy. Cancer cells may acquire resistance to chemotherapy through a range of mechanisms, including mutation or overexpression of drug targets, inactivation of drugs, or elimination of drugs from cells.
In one embodiment, the invention provides a pharmaceutical composition comprising an ALK inhibitor, and one or more anticancer reagents selected from a CDK4/6 inhibitor, an Mek inhibitor, a chemotherapeutic agent, an MDM2 inhibitor, an HDAC inhibitor, a PD-1 inhibitor, a PARP inhibitor, a VEGF inhibitor and a BCR-ABL inhibitor.
In a preferred embodiment, the ALK inhibitor is a compound of Formula I or a pharmaceutically acceptable salt or solvate thereof:
In a preferred embodiment, the ALK inhibitor is a compound of Formula II or a pharmaceutically acceptable salt or solvate thereof:
In a preferred embodiment, the ALK inhibitor is a compound of Formula III or a pharmaceutically acceptable salt or solvate thereof:
In a preferred embodiment, the ALK inhibitor is a compound of Formula IV or a pharmaceutically acceptable salt or solvate thereof:
In some embodiments, the compound has an enantiomeric excess of about 91% or more, about 92% or more, about 93% or more, about 94% or more, about 95% or more, about 96% or more, about 97% or more, about 98% or more, or about 99% or more.
In a preferred embodiment, the ALK inhibitor is a compound of Formula V or a pharmaceutically acceptable salt or solvate thereof:
In a preferred embodiment, the ALK inhibitor is a compound of Formula VI or a pharmaceutically acceptable salt or solvate thereof:
In a preferred embodiment, the ALK inhibitor is a compound in the following table or a pharmaceutically acceptable salt or solvate thereof:
In a preferred embodiment, the ALK inhibitor is 5-chloro-N2-(2-isopropoxy-5 -methyl-4-(1-(tetrahydro-2H-pyran-4-yl)- 1,2,3,6-t etrahydropyridin-4-yl)phenyl)-N4-(2-(isopropylsulfonyl)phenyl)pyrimidine-2,4-diamine, or a pharmaceutically acceptable salt or hydrate thereof.
In a preferred embodiment, the CDK4/6 inhibitor is compound of the formulae (VII) or (VIII), or pharmaceutically acceptable salt thereof:
In a preferred embodiment, the CDK4/6 inhibitor is palbociclib, ribociclib or a pharmaceutically acceptable salt thereof.
In a preferred embodiment, the Mek inhibitor is compound of the formulae (IX) or (X), or pharmaceutically acceptable salt thereof:
In a preferred embodiment, the Mek inhibitor is trametinib, selumetinib or a pharmaceutically acceptable salt thereof.
In a preferred embodiment, the chemotherapeutic agent comprises platinum or belongs to terpenoid alkaloid.
In a preferred embodiment, the chemotherapeutic agent is carboplatin or paclitaxel.
In a preferred embodiment, the MDM2 inhibitor is compound of the formula (XI) or pharmaceutically acceptable salt thereof:
is
is hydrogen, CH3, CH2CH3, C3 alkyl or C4 alkyl;
In a preferred embodiment, the MDM2 inhibitor is Compound 33 with the following structure including any tautomer forms, or a pharmaceutically acceptable salt thereof:
In a preferred embodiment, the HDAC inhibitor is compound of the formula (XII) or pharmaceutically acceptable salt thereof:
In a preferred embodiment, the HDAC inhibitor is panobinostat or a pharmaceutically acceptable salt thereof.
In a preferred embodiment, the PD-1 inhibitor is anti-PD-1 antibody.
In a preferred embodiment, the PARP inhibitor is compound of the formula (XIII) or pharmaceutically acceptable salt thereof:
In a preferred embodiment, the PARP inhibitor is Olaparib or a pharmaceutically acceptable salt thereof.
In a preferred embodiment, the VEGF inhibitor is compound of the formula (XIV) or pharmaceutically acceptable salt thereof:
In a preferred embodiment, the VEGF inhibitor is Lenvatinib or a pharmaceutically acceptable salt thereof.
In a preferred embodiment, the BCR-ABL inhibitor is compound of the formula (XV) or pharmaceutically acceptable salt thereof:
In a preferred embodiment, the BCR-ABL inhibitor is Compound 34 with the following structure including any tautomer forms, or a pharmaceutically acceptable salt thereof:
In a preferred embodiment, the pharmaceutical composition is for treating or suppressing a cancer, reducing its severity, lowering its risk or inhibiting its metastasis in an individual, and the cancer is selected from the group consisting of bladder cancer, breast cancer, cervical cancer, colon cancer (including colorectal cancer), esophageal cancer, esophageal squamous cell carcinoma, head and neck cancel; liver cancer, lung cancer (including small cell lung cancer and non-small cell lung cancer), melanoma, myeloma, rhabdomyosarcoma, inflammatory myofibroblastic tumor, neuroturbo chargeoma, pancreatic cancer, prostate cancel; kidney cancer, renal cell carcinoma, sarcoma (including osteosarcoma), skin cancer (including squamous cell carcinoma), gastric cancer, testicular cancer, thyroid cancer, uterine cancel; mesothelioma, neuroblastoma, cholangiocarcinoma, leiomyosarcoma, lipo sarcoma, nasopharyngeal carcinoma, neuroendocrine carcinoma, ovarian cancer, salivary gland cancer, metastasis caused by spindle cell carcinoma, anaplastic large cell lymphoma, thyroid undifferentiated carcinoma, non-Hodgkin's lymphoma, Hodgkin's lymphoma, and hematological malignancies, such as acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), uveal melanoma, glioblastoma;
In a preferred embodiment, the weight ratio between the ALK inhibitor and the one or more anticancer reagents is 0.005-5000: 0.005-5000, for example, 0.05-1500:0.005-5000, 0.1-6:0.005-4, 100:0.5-400, 100:1-350, 100:2-300, 100:5-200, 100:10-150, 100:20-100, 100:30-90, 100:20-80.
In some preferred embodiment, the cancer is mesothelioma, neuroblastoma, non-small cell lung cancer, lung adenocarcinoma (LUAD), ovarian cancer, uveal melanoma, glioblastoma, colon cancer, and liver cancer.
In a preferred embodiment, the ALK inhibitor is administrated in an amount of from about 0.005 mg/day to about 5000 mg/day, such as an amount of about 0.005, 0.05, 0.5, 5, 10, 20, 30, 40, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500 or 5000 mg/day.
In a preferred embodiment, the ALK inhibitor is administrated in an amount of from about 1 ng/kg to about 200 mg/kg, about 1 μg/kg to about 100 mg/kg, or about 1 mg/kg to about 50 mg/kg per unit dose, for example, administrated in an amount of about 1 μg/kg, about 10 μg/kg, about 25 μg/kg, about 50 μg/kg, about 75 μg/kg, about 100 μg/kg, about 125 μg/kg, about 150 μg/kg, about 175 μg/kg, about 200 μg/kg, about 225 μg/kg, about 250 μg/kg, about 275 μg/kg, about 300 μg/kg, about 325 μg/kg, about 350 μg/kg, about 375 μg/kg, about 400 μg/kg, about 425 μg/kg, about 450 μg/kg, about 475 μg/kg, about 500 μg/kg, about 525 μg/kg, about 550 μg/kg, about 575 μg/kg, about 600 μg/kg, about 625 μg/kg, about 650 μg/kg, about 675 μg/kg, about 700 μg/kg, about 725 μg/kg, about 750 μg/kg, about 775 μg/kg, about 800 μg/kg, about 825 μg/kg, about 850 μg/kg, about 875 μg/kg, about 900 μg/kg, about 925 μg/kg, about 950 μg/kg, about 975 μg/kg, about 1 mg/kg, about 5 mg/kg, about 10 mg/kg, about 15 mg/kg, about 20 mg/kg, about 25 mg/kg, about 30 mg/kg, about 35 mg/kg, about 40 mg/kg, about 45 mg/kg, about 50 mg/kg, about 60 mg/kg, about 70 mg/kg, about 80 mg/kg, about 90 mg/kg, about 100 mg/kg, about 125 mg/kg, about 150 mg/kg, about 175 mg/kg, about 200 mg/kg per unit dose, and administrated with one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) unit doses per day.
In a preferred embodiment, the one or more anticancer reagents are administrated in an amount of from 0.005 mg/day to about 5000 mg/day, for example, about 0.005, 0.05, 0.5, 5, 10, 20, 30, 40, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500 or 5000 mg/day.
In a preferred embodiment, the one or more anticancer reagents are administrated in an amount of from about 1 ng/kg to about 200 mg/kg, from about 1 μg/kg to about 100 mg/kg, or from about 1 mg/kg to about 50 mg/kg per unit dose, for example, administrated in an amount of about 1 μg/kg, about 10 μg/kg, about 25 μg/kg, about 50 μg/kg, about 75 μg/kg, about 100 μg/kg, about 125 μg/kg, about 150 μg/kg, about 175 μg/kg, about 200 μg/kg, about 225 μg/kg, about 250 μg/kg, about 275 μg/kg, about 300 μg/kg, about 325 μg/kg, about 350 μg/kg, about 375 μg/kg, about 400 μg/kg, about 425 μg/kg, about 450 μg/kg, about 475 μg/kg, about 500 μg/kg, about 525 μg/kg, about 550 μg/kg, about 575 μg/kg, about 600 μg/kg, about 625 μg/kg, about 650 μg/kg, about 675 μg/kg, about 700 μg/kg, about 725 μg/kg, about 750 μg/kg, about 775 μg/kg, about 800 μg/kg, about 825 μg/kg, about 850 μg/kg, about 875 μg/kg, about 900 μg/kg, about 925 μg/kg, about 950 μg/kg, about 975 μg/kg, about 1 mg/kg, about 5 mg/kg, about 10 mg/kg, about 15 mg/kg, about 20 mg/kg, about 25 mg/kg, about 30 mg/kg, about 35 mg/kg, about 40 mg/kg, about 45 mg/kg, about 50 mg/kg, about 60 mg /kg, about 70 mg/kg, about 80 mg/kg, about 90 mg/kg, about 100 mg/kg, about 125 mg/kg, about 150 mg/kg, about 175 mg/kg, about 200 mg/kg per unit dose, and administered with one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) unit doses per day.
In a preferred embodiment, the ALK inhibitor, and the one or more anticancer reagents are administered together, simultaneously, sequentially or alternately.
In a preferred embodiment, the ALK inhibitor, and the one or more anticancer reagents are administered continuously for at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, at least 11 days, at least 12 days, at least 13 days, at least 14 days, at least 15 days, at least 16 days, at least 17 days, at least 18 days, at least 19 days, at least 20 days, at least 21 days, at least 22 days, at least 23 days, at least 24 days, at least 25 days, at least 30 days, at least 35 days, at least 40 days, at least 45 days, or at least 50 days.
In a preferred embodiment, the ALK inhibitor, and the one or more anticancer reagents are administered for one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) courses of treatment, in which each of the courses lasts at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, at least 11 days, at least 12 days, at least 13 days, at least 14 days, at least 15 days, at least 16 days, at least 17 days, at least 18 days, at least 19 days, at least 20 days, at least 21 days, at least 22 days, at least 23 days, at least 24 days, at least 25 days, at least 30 days, at least 35 days, at least 40 days, at least 45 days, or at least 50 days; and there is an interval of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 days, two weeks, three weeks or four weeks between every two courses of treatment.
In a preferred embodiment, when there are a plurality of courses of treatment, the amount of the ALK inhibitor and/or anticancer reagents administered in each course of treatment is same or different. In a more preferred embodiment, the amount of the ALK inhibitor and/or anticancer reagents administered during the previous course of treatment is 1-10 times, preferably 1-5 times, such as 1.5, 2, 2.5, 3, 3.5, 4, 4.5 or 5 times, the amount administered during the subsequent course of treatment.
In a preferred embodiment, the ALK inhibitor, and the one or more anticancer reagents are administrated via the same (e.g., oral) or different routes (e.g., oral and parenteral (e.g., injection), respectively).
In a preferred embodiment, the anticancer reagent is administrated in a lower dose in comparison with the dose of the anticancer reagent that is administered alone or when the one or more ALK inhibitors are not administered.
In a preferred embodiment, the ALK inhibitor enhances the therapeutic efficacy of the anticancer reagent in treatment of a cancer and/or reduces a side-effect of the anticancer reagent in treatment of a cancer.
In a preferred embodiment, the invention provides a use of an ALK inhibitor in manufacture of a medicament for enhancing the efficacy of an anticancer reagent in treatment of a cancer and/or reducing a side-effect of an anticancer reagent in treatment of a cancer.
In a preferred embodiment, the individual suffers from an advanced cancer.
In a preferred embodiment, the individual suffers from a refractory cancer, a recurrent cancer or a drug-resistant cancer.
In another embodiment, the invention provides a method for treating or suppressing a cancer, reducing its severity, lowering its risk or inhibiting its metastasis in an individual, comprising administering to the individual a therapeutically effective amount of an ALK inhibitor, and a therapeutically effective amount of one or more anticancer reagents selected from a CDK4/6 inhibitor, an Mek inhibitor, a BRAF inhibitor a chemotherapeutic agent, an MDM2 inhibitor, an HDAC inhibitor, a PD-1 inhibitor, a PARP inhibitor, a VEGF inhibitor and a BCR-ABL inhibitor;
In a preferred embodiment, the ALK inhibitor is as defined above and the anticancer reagent is as defined above.
In another embodiment, the invention provides a use of a pharmaceutical composition in manufacture of a medicament for treating or suppressing a cancer, reducing its severity, lowering its risk or inhibiting its metastasis in an individual, the pharmaceutical composition comprising an ALK inhibitor, and one or more anticancer reagent, as well as optionally a pharmaceutically acceptable carrier;
In a preferred embodiment, the ALK inhibitor is as defined above and the anticancer reagent is as defined above.
In another embodiment, the invention provides a kit, comprising:
In order to make the objects and technical solutions of the present invention clearer, the present invention will be further described below in conjunction with specific example. It should be understood that the examples are not intended to limit the scope of the invention. Further, specific experimental methods not mentioned in the following examples were carried out in accordance with a conventional experimental method.
The compounds of formulae (I) to (VI) or a pharmaceutically acceptable salt thereof including Compound 5 (5-chloro-N2-(2-isopropoxy-5-methyl-4-(1-(tetrahydro-2H-pyran-4-yl)-1,2,3,6-tetrahydropyridin-4-yl)phenyl)-N4-(2-(isopropylsulfonyl)phenyl)pyrimidine-2,4-diamine) are synthesized according to the production methods described in WO 2018/044767, which is incorporated herein by reference in its entirety and for all purposes, or a method analogous thereto.
The compounds of formula (XV) or a pharmaceutically acceptable salt thereof including Compound 34 can be obtained according to the production methods described in U.S. Pat. No. 8,846,671 B2, issued Sep. 30, 2014, which is incorporated herein by reference in its entirety and for all purposes, or a method analogous thereto.
Cell plating: Anti-proliferative effects were detected by a CCK-8 (Cell Counting Kit-8, Shanghai life iLab, China) assay based on water soluble tetrazolium salt (WST). The cells were seeded in 96-well plates, and only 95 μL of complete medium was added to each negative control group. 95 μL of complete medium cell suspension was added to each well to be tested, and the cell density was (5-10)×10{circumflex over ( )}4/hole.
Dosing (protection from light): In 96-well culture plates, according to the sensitivity of different cells to different drugs, the highest concentration was selected as 3.7 μM, and 6 concentrations were obtained by serial dilution in a ratio of 1:3. 5 μL of compound was added to each well and 2-3 replicate wells were made for per concentration. After the compound was added, 96-well plates were incubated in a 5% CO2 incubator at 37° C. After 72 hours of action by using 6 different concentrations of Compound 34 with 3 fixed doses of Compound 5, the combination effect of Compound 5 an Compound 34 was tested.
Reading: At the end of the culture, the old solution was removed from the well to be tested, and 100 μl/well CCK-8 test solution (containing 10% CCK-8, 5% FBS in the corresponding medium) was added. The plates were continuously incubated at 37 ° C. for 2-4 hours in a CO2 incubator.
The OD values were measured using a microplate reader (SpectraMax Plus 384, Molecular Devices, LLC., US) under A450 nm. Using the average OD value of 3 replicate wells, the percentage of cell viability was calculated by the following formula:
(O.D. of test well−O.D. of blank control well)/(O.D. of cell control well−O.D. of blank control well)×100%.
IC50 values were calculated using Graphpad Prism 6.0 software for nonlinear regression data analysis method. The results are shown in
For combination experiments, cell viability was calculated by normalization of the mean OD values of 3 replicate wells of single drug control. The comparison of the IC50 values obtained from the curves of combined drugs of administration and single drug of administration shows that the two compounds achieved synergistic effect (the curve of the combined drugs of administration shifted to the left).
As shown in Table 1, Compound 34 alone or a combination of Compound 5 and Compound 34 was used to treat NCI-H2228 cells for 72 hours, and such combination shows a synergistic antiproliferative activity in this model.
The compounds of formula (XI) or a pharmaceutically acceptable salt thereof including Compound 33 and any tautomer forms was prepared using the one or more procedures described in U.S. Pat. No. 9745314, and Aguilar et al. J. Med. Chem. 2017(60), 2819-2839.
Cell viability was determined using CellTiter-Glo® luminescent cell viability assay (Promega) by following manufacturer's instruction. Cell viability was calculated as cell viability=(mean RLU sample−mean RLU blank)/(RLU cell control−RLU blank)×100. IC50 value was calculated using GraphPad Prism. Combination index (CI) value was calculated by CalcuSyn software (BIOSOFT, UK). CI<0.9 indicate a synergistic combination effect. CI<0.1 scored as 5+ indicates very strong synergistic combination effect, CI between 0.1 and 0.3 scored as 4+ indicates strong synergistic combination effect, CI between 0.3 and 0.7 scored as 3+ indicates medium synergistic combination effect.
Uveal melanoma MP41 cells (ATCC) were treated with Compound 5, MDM2 inhibitor Compound 33 or a combination of Compound 5 and Compound 33 respectively, wherein the culture of RPMI 1640+10% FBS (Fetal Bovine Serum)+1% P/S (Penicillin-Streptomycin) was used.
The results are shown in
A subcutaneous xenograft tumor model of human tumor immunodeficient mice was established by cell inoculation: tumor cells in logarithmic growth phase were collected, counted, resuspended in 1×PBS, and the cell suspension concentration was adjusted to 2.5-5 ×107/mL. The tumor cells were inoculated subcutaneously in the right side of immunodeficient mice with a 1 mL syringe (4 gauge needle), 5-10×106/0.2 mL/mouse. All animal experiments were strictly in accordance with the specifications for the use and management of experimental animals in GenePharma Co., Ltd. and Suzhou Ascentage Pharma Co., Ltd. The calculation of relevant parameters refers to the Chinese NMPA “Guidelines for Non-Clinical Research Techniques of Cytotoxic Anti-tumor Drugs”.
Animal body weight and tumor size were measured twice weekly during the experiment. The state of the animal and the presence or absence of death were observed every day. The growth of tumor and the effects of treatment on normal behavior of animals were monitored routinely, specifically involving experimental animal activity, feeding and drinking, weight gain or loss, eyes, clothing hair and other abnormalities. The deaths and clinical symptoms observed during the experiment were recorded in the raw data. All operations for administration and measurement of mouse body weight and tumor volume were performed in a clean bench. According to the requirements of the experimental protocol, after the end of the last administration, plasma and tumor tissues were collected, weighed and photographed. The plasma and tumor samples were frozen at −80° C. for ready-to-use.
Tumor volume (TV) is calculated as: TV=a×b2/2, wherein a and b represent the length and width of the tumor to be measured, respectively.
The relative tumor volume (RTV) is calculated as: RTV=Vt/V1, wherein V1 is the tumor volume at the start of grouping and administration, and Vt is the tumor volume measured on the t day after administration.
The evaluation index of anti-tumor activity is the relative tumor proliferation rate T/C (%), and the calculation formula thereof is: relative tumor proliferation rate T/C (%)=(TRTV/CRTV)×100%, TRTV is the RTV of treatment group, CRTV is the RTV of solvent control group.
Tumor regression rate (%) is calculated as: the number of tumor-bearing mice which exhibit SD (stable disease), PR (partial regression) and CR (complete regression) after treatment/the total number of the mice in this group×100%.
Change of body weight (%)=(measured body weight−body weight at the start of grouping)/body weight at the start of grouping×100%.
Evaluation criteria for therapeutic efficiency: According to the Chinese NMPA “Guidelines for Non-Clinical Research Techniques of Cytotoxic Anti-tumor Drugs” (November 2006), when T/C (%) value is <40% and statistical analysis shows p<0.05, efficiency is confirmed. A dose of drug is considered to be severely toxic if the body weight of mouse is reduced by more than 20% or the number of drug-related deaths exceeds 20%.
According to the description by Clarke R., Issues in experimental design and endpoint analysis in the study of experimental cytotoxic agents in vivo in breast cancer and other models [J]. Breast Cancer Research & Treatment, 1997, 46(2-3): 255-278, synergy analysis was evaluated using the following formula: synergy factor=((A/C)×(B/C))/(AB/C); A=RTV value of drug A alone group; B=RTV value of drug B alone group; C=RTV value of the solvent control group, and AB=RTV value of the A and B combination group. Synergistic factor>1 indicates that synergy is achieved; synergy factor=1 indicates that additive effect is achieved; and synergy factor<1 indicates that antagonistic effect is achieved.
Use of mRECIST (Gao et al., 2015) measured tumor responses included stable disease (SD), partial tumor regression (PR), and complete regression (CR), determined by comparing tumor volume change at day t to its baseline: tumor volume change (%)=(Vt−V1)/V1. The BestResponse was the minimum value of tumor volume change (%) for t≥10. For each time t, the average of tumor volume changes from t=1 to t was also calculated. BestAvgResponse was defined as the minimum value of this average for t≥10. The criteria for response (mRECIST) were adapted from RECIST criteria (Gao et al., 2015; Therasse et al., 2000) and defined as follows: mCR, BestResponse<−95% and BestAvg Response<−40%; mPR, BestResponse<−50% and BestAvgResponse<−20%; mSD, BestResponse<35% and BestAvgResponse<30%; mPD, not otherwise categorized. SD, PR, and CR were considered responders and used to calculate response rate (%). Disease control rate (DCR) is calculated with the proportion of animals demonstrating CR, PR, or SD based on mRECIST; Overall response rate (ORR) is calculated with the proportion of animals demonstrating CR or PR based on mRECIST. Body weight of animals were monitored simultaneously. The change in body weight was calculated based on the animal weight of the first day of dosing (day 1). Tumor volume and changes in body weight (%) were represented as the mean±standard error of the mean (SEM).
The evaluation method as described in Example 3 is used in Examples 4-20.
In this experiment, a human MT16036 cell-derived xenograft (TP53 mut, FAK amp and CDK4 high) tumor model (Crown Bioscience) was established to evaluate the anti-tumor effect of Compound 5 in combination with CDK4/6 inhibitor palbociclib (Yishiming(Beijing) Pharm-Chemicals Tech. Co., Ltd). The dosing regimen was as follows:
The dosing regimen of each drug in the dosing regimen for the combination is the same as the dosing regimen for the single drug.
As shown in Table 2 and
In addition, as shown in
In conclusion, Compound 5 single agent showed minor antitumor activity, palbociclib single agent showed moderate antitumor activity, while the combination treatment showed enhanced antitumor activity, synergistic antitumor effect and acceptable toxicity in s.c. Mesothelioma PDX.
In this experiment, an Neuroblastoma SH-SY5Y (ALK F1174L) xenograft tumor model was established to evaluate the anti-tumor effect of Compound 5 in combination with CDK4/6 inhibitor ribociclib (Yishiming(Beijing) Pharm-Chemicals Tech. Co., Ltd). The dosing regimen was as follows:
The dosing regimen of each drug in the dosing regimen for the combination is the same as the dosing regimen for the single drug.
The cell source is ATCC, and the cell culture is F12K: MEM=1:1 with sodium pyruvate (1×) and NEAA (1×), 90%; fetal bovine serum, 10%; P/S, 1%.
As shown in Table 3 and
In addition, as shown in
In conclusion, ribociclib single agent showed moderate antitumor activity, Compound 5 single agent showed strong antitumor activity, while the combination treatment showed significantly enhanced tumor regression, strong synergistic antitumor effect and acceptable toxicity in s.c. Neuroblastoma SH-SY5Y xenograft.
†p < 0.05, compared to Compound 5 group, §p < 0.05, compared to ribociclib group
In this experiment, a human A549 cell-derived xenograft lung tumor model was established to evaluate the anti-tumor effect of Compound 5 in combination with MEK inhibitor trametinib/selumetinib (Selleck). The dosing regimen was as follows:
The dosing regimen of each drug in the dosing regimen for the combination is the same as the dosing regimen for the single drug.
The cell source is Cobioer, and the cell culture is RPMI 1640 medium with 300 mg/L (2 mM) L-glutamine adjusted to contain 2.0 g/L sodium bicarbonate, 90%; fetal bovine serum, 10%; P/S 1%.
As shown in Table 4 and
In addition, as shown in
In conclusion, in A549 NSCLC xenograft, Compound 5 single agent or MEK inhibitor (trametinib or selumetinib) showed minor antitumor activity, while combination treatment of Compound 5 and MEK inhibitor (trametinib or selumetinib) showed significantly enhanced antitumor activity and synergistic antitumor effect in A549 NSCLC xenograft.
In this experiment, a PTK2 high NSCLC PDX model of LU-01-0604 (Wuxi Pharma Tech) was established to evaluate the anti-tumor effect of Compound 5 in combination with chemotherapeutic agent carboplatin (Selleck). The dosing regimen was as follows:
The dosing regimen of each drug in the dosing regimen for the combination is the same as the dosing regimen for the single drug.
As shown in Table 5 and
After 29 days of administration, the combination group had a synergistic factor of 1.92, indicating synergistic effects.
In addition, as shown in
In conclusion, Compound 5 single agent and carboplatin single agent showed no antitumor activity, while the combination treatment showed enhanced antitumor activity and synergistic antitumor effect in s.c. NSCLC PDX.
In this experiment, an ovarian PA-1 xenograft tumor model was established to evaluate the anti-tumor effect of Compound 5 in combination with chemotherapeutic agent carboplatin (Selleck). The dosing regimen was as follows:
The dosing regimen of each drug in the dosing regimen for the combination is the same as the dosing regimen for the single drug.
The cell source is Cobioer, and the cell culture is MEM medium with fetal bovine serum of 10% and P/S of 1%.
As shown in Table 6 and
In addition, as shown in
In conclusion, Compound 5 single agent showed moderate antitumor activity, while the combination treatment showed synergistic antitumor effect in s.c. PA-1 ovarian cancer xenografts.
+p < 0.05 compared to Compound 5 group;
#p < 0.05 compared to carboplatin group
In this experiment, a PTK2 high ovarian PDX model of OV2423 was established to evaluate the anti-tumor effect of Compound 5 in combination with chemotherapeutic agent carboplatin and/or paclitaxel (Harbin Pharmaceutical Group). The dosing regimen was as follows:
The dosing regimen of each drug in the dosing regimen for the combination is the same as the dosing regimen for the single drug.
As shown in Table 7,
As shown in Table 7,
In addition, as shown in
In conclusion, Compound 5 single agent showed minor antitumor activity, carboplatin single agent or paclitaxel single agent showed moderate antitumor activity, while combination treatment of Compound 5 and paclitaxel or combination treatment of Compound 5 and carboplatin achieved synergistic antitumor effects in s.c. ovarian cancer PDX model.
In this experiment, an ovarian PDX model of OV1658 was established to evaluate the anti-tumor effect of Compound 5 in combination with chemotherapeutic agent carboplatin and/or paclitaxel (Harbin Pharmaceutical Group). The dosing regimen was as follows:
The dosing regimen of each drug in the dosing regimen for the combination is the same as the dosing regimen for the single drug.
As shown in Table 8,
As shown in Table 8,
In conclusion, carboplatin single agent or paclitaxel single agent showed minor antitumor activity, while combination treatment of Compound 5 and paclitaxel or combination treatment of Compound 5 and paclitaxel plus carboplatin achieved synergistic antitumor effects in s.c. ovarian cancer PDX model.
In this experiment, a PTK2 high ovarian PDX model of OV1385 was established to evaluate the anti-tumor effect of Compound 5 in combination with chemotherapeutic agent carboplatin and/or paclitaxel (Harbin Pharmaceutical Group). The dosing regimen was as follows:
The dosing regimen of each drug in the dosing regimen for the combination is the same as the dosing regimen for the single drug.
As shown in Table 9,
As shown in Table 9,
In addition, as shown in
In conclusion, carboplatin single agent or paclitaxel single agent showed moderate antitumor activity, while combination treatment of Compound 5 and paclitaxel or combination treatment of Compound 5 and paclitaxel plus carboplatin achieved synergistic antitumor effects in s.c. ovarian cancer PDX model.
In this experiment, a PTK2 high ovarian PDX model of OV2018 was established to evaluate the anti-tumor effect of Compound 5 in combination with chemotherapeutic agent paclitaxel (Harbin Pharmaceutical Group). The dosing regimen was as follows:
The dosing regimen of each drug in the dosing regimen for the combination is the same as the dosing regimen for the single drug.
As shown in Table 10 and
In conclusion, Compound 5 singe agent and paclitaxel single agent showed no antitumor activity, while combination treatment of Compound 5 and paclitaxel achieved a synergistic antitumor effect in s.c. ovarian cancer PDX model.
In this experiment, an OVCAR3 subcutaneous ovarian xenograft model was established to evaluate the anti-tumor effect of Compound 5 in combination with chemotherapeutic agent paclitaxel and carboplatin (Selleck). The dosing regimen was as follows:
The dosing regimen of each drug in the dosing regimen for the combination is the same as the dosing regimen for the single drug.
The cell source is China Center for Type Culture Collection (CCTCC), and the cell culture is RPMI 1640 medium with 300 mg/L (2 mM) L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate, 4.5 g/L glucose, 10 mM HEPES, and 1.0 mM sodium pyruvate, 80%; fetal bovine serum, 20%; P/S 1%.
As shown in Table 11 and
In addition, as shown in
In conclusion, Compound 5 single agent showed minor antitumor activity, and paclitaxel single agent showed moderate antitumor activity, while the combination treatment of Compound 5 and paclitaxel or the combination treatment of Compound 5 and paclitaxel plus carboplatin achieved synergistic antitumor effects in s.c. ovarian cancer PDX model.
In this experiment, an OVCAR3 subcutaneous ovarian xenograft model was established to evaluate the activity of Compound 5 in overcoming the resistance to chemotherapeutic agent paclitaxel and carboplatin (Selleck). The dosing regimen was as follows:
Paclitaxel (10 mg/kg) was intravenously injected to the mice once a week for the first 14 days (n=8). Randomize the mice into two groups on Day 15, one group (n=5) was continuously intravenously injected for paclitaxel single agent (10 mg/kg, once a week) until Day 56, and the other group (n=3) was treated with the combination of paclitaxel (10 mg/kg, once a week) and Compound 5 (100 mg/kg, orally, once per day) from Day 15 until Day 56. The results are shown in Table 12 and
Paclitaxel (10 mg/kg, intravenous injection, once a week) and carboplatin (30 mg/kg, intravenous injection, once a week) combination treatment was used for the mice for the first 14 days (n=12). Randomize the mice into three groups on Day 15, one group (n=5) continuously underwent the same amount of paclitaxel and carboplatin combination treatment until Day 56, another group (n=4) underwent paclitaxel plus carboplatin and Compound 5 (100 mg/kg, orally, once per day) combination treatment, and the third group (n=3) underwent Compound 5 single agent treatment. The results are shown in Table 12 and
The cell source and cell culture are the same with Example 13.
As shown in Table 12 and
As shown in Table 12 and
In this experiment, a s.c. LUAD A549 (KRAS, STK11, ATR mut) xenograft model was established to evaluate the anti-tumor effect of Compound 5 in combination with Compound 33. The dosing regimen was as follows:
The dosing regimen of each drug in the dosing regimen for the combination is the same as the dosing regimen for the single drug.
The cell source is Cobioer, and the cell culture is RPMI 1640 medium with 300 mg/L (2 mM) L-glutamine adjusted to contain 2.0 g/L sodium bicarbonate, 90%; fetal bovine serum, 10%; P/S 1%.
As shown in Table 13 and
In addition, as shown in
In conclusion, Compound 5 single agent and Compound 33 single agent showed minor antitumor activity, while combination treatment of Compound 5 and Compound 33 achieved a synergistic antitumor effect in s.c. A549 NSCLC xenograft model.
In this experiment, a s.c. Neuroblastoma SH-SY5Y (ALK F1174L) xenograft model was established to evaluate the anti-tumor effect of Compound 5 in combination with Compound 33. The dosing regimen was as follows:
The dosing regimen of each drug in the dosing regimen for the combination is the same as the dosing regimen for the single drug.
The cell source is ATCC, and the cell culture is F12K: MEM=1:1 with sodium pyruvate (1×) and NEAA (1×), 90%; fetal bovine serum, 10%; P/S, 1%.
As shown in Table 14 and
In addition, as shown in
In conclusion, Compound 5 single agent and Compound 33 single agent showed strong antitumor activity, and combination treatment of Compound 5 and Compound 33 achieved an enhanced antitumor effect in s.c. SH-SYSY Neuroblastoma xenograft model, achieved ORR 80% compared to 0 in other groups.
In this experiment, a subcutaneous A549 lung cancer model was established to evaluate the anti-tumor effect of Compound 5 in combination with HDAC inhibitor panobinostat. The dosing regimen was as follows:
The dosing regimen of each drug in the dosing regimen for the combination is the same as the dosing regimen for the single drug.
The cell source is Cobioer, and the cell culture is RPMI 1640 medium with 300 mg/L (2 mM) L-glutamine adjusted to contain 2.0 g/L sodium bicarbonate, 90%; fetal bovine serum, 10%; P/S 1%.
As shown in Table 15 and
In conclusion, Compound 5 single agent showed minor antitumor activity, panobinostat single agent showed moderate antitumor activity, while combination treatment of Compound 5 and panobinostat achieved a synergistic antitumor effect in s.c. A549 NSCLC xenograft model.
In this experiment, a s.c. syngeneic tumor model of colon CT26 was established to evaluate the anti-tumor effect of Compound 5 in combination with anti-PD-1 antibody (Bioxcell, Item No. BE0146, Clone No: RMP1-14). The dosing regimen was as follows:
The dosing regimen of each drug in the dosing regimen for the combination is the same as the dosing regimen for the single drug.
The cell source is Cobioer, and the cell culture is RPMI 1640+10% FBS+1% P/S.
As shown in Table 16 and
In addition, as shown in
In conclusion, Compound 5 single agent or anti-PD-1 single agent showed no antitumor activity, while the combination treatments achieved enhanced antitumor activities and synergistic antitumor effect in s.c. CT26 syngeneic colon cancer tumor model.
In this experiment, a s.c. FAK-high NSCLC LU-01-0751 PDX (BRCA2 mut) model (Wuxi Pharma Tech) was established to evaluate the anti-tumor effect of Compound 5 in combination with PARP inhibitor Olaparib (Selleck). The dosing regimen was as follows:
The dosing regimen of each drug in the dosing regimen for the combination is the same as the dosing regimen for the single drug.
As shown in Table 17 and
In conclusion, Compound 5 single agent or Olaparib single agent showed no antitumor activity, while the combination treatment with Compound 5 and Olaparib achieved a synergistic antitumor effect in s.c. NSCLC PDX model.
In this experiment, a PTK2-high liver PDX model of LI-03-1140 (Wuxi Pharma Tech) was established to evaluate the anti-tumor effect of Compound 5 in combination with VEGF inhibitor Lenvatinib (Selleck). The dosing regimen was as follows:
The dosing regimen of each drug in the dosing regimen for the combination is the same as the dosing regimen for the single drug.
As shown in Table 18 and
In conclusion, Compound 5 single agent showed no antitumor activity, Lenvatinib single agent showed strong antitumor activity, and the combination treatment achieved a synergistic antitumor effect in s.c. liver cancer PDX model.
In this experiment, a TP53wt , BRAFv600E , NRASwt, PTENwt, CDKN2Amut C32 cutaneous melanoma model (Wuxi Pharma Tech) was established to evaluate the anti-tumor effect of Compound 5 in combination with BRAFi (Dabrafenib)+MEKi (trametinib) (Selleck). The dosing regimen was as follows:
The dosing regimen of each drug in the dosing regimen for the combination is the same as the dosing regimen for the single drug.
As shown in Table19 and
In conclusion, Compound 5 single agent showed no antitumor activity, Dabrafenib +trametinib showed strong antitumor activity, and the combination treatment achieved a synergistic antitumor effect in TP53wt, BRAFv600E, NRASwt, PTENwt, CDKN2Amut C32 cutaneous melanoma model.
In this experiment, a U-87-MG subcutaneous glioblastoma model (Wuxi Pharma Tech) was established to evaluate the anti-tumor effect of Compound 5 in combination with Palbociclib/Trametinib/TMZ. The dosing regimen was as follows:
The dosing regimen of each drug in the dosing regimen for the combination is the same as the dosing regimen for the single drug.
As shown in Table 20 and
###P < 0.001, vs. COMPOUND 5;
&&&P < 0.001, vs. Palbociclib;
$P < 0.05, vs. Trametinib;
In conclusion, Compound 5 single agent showed no antitumor activity, and the combination treatment achieved a synergistic antitumor effect in U-87-MG subcutaneous glioblastoma model.
(B/C))/(AB/C); A =
100%; TRTV is RTV of the
| Number | Date | Country | Kind |
|---|---|---|---|
| PCT/CN2020/117895 | Sep 2020 | WO | international |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/CN2021/120260 | 9/24/2021 | WO |
