Pharmaceutical composition for cancer treatment which contains nanoparticles containing slightly soluble camptothecin compound, and combination therapy of the same

Abstract

To provide a pharmaceutical composition for cancer treatment which shows a synergistic cancer therapeutic effect when used in combination with an anti-tumor agent, and can be easily used as a cancer therapeutic agent and a combination therapeutic pharmaceutical.SOLUTION: Provided is a cancer treatment application of nanoparticles containing a slightly soluble camptothecin-based compound and a combined administration therapy with an anti-tumor agent using the application. In one embodiment, a combination pharmaceutical preparation for pancreatic cancer treatment contains: particles including SN-38(7-ethyl-10-hydroxycamptothecin), irinotecan, and an amphiphilic block copolymer made of a hydrophobic block and a hydrophilic block; and one or more kinds of anti-tumor agents selected from the group consisting of nab-paclitaxel as an active ingredient, the amphiphilic block copolymer being constituted of A-B or A-B-A block, (a) A is a hydrophilic polymer and is monomethoxypolyethylene, and (b) B is a hydrophobic polymer and is poly-D,L-lactide or a derivative thereof.SELECTED DRAWING: Figure 2

Description

[Technical field]

[0001] This patent application claims priority to Korean Patent Application No. 10-2020-0163156, filed with the Korean Intellectual Property Office on November 27, 2020, the disclosure of which is incorporated herein by reference.

[0002] The present invention relates to a pharmaceutical composition for treating cancer, comprising nanoparticles containing a poorly soluble camptothecin compound, and to combination therapy thereof.

[0003] [Background technology]

[0004] Cancer treatment methods are divided into surgery, radiation therapy, and drug therapy (anticancer chemotherapy or anticancer drug therapy), but since surgery, the primary treatment method, is not possible for malignant tumors, anticancer chemotherapy (single or combined) is selected. Cytotoxic anticancer drugs used in chemotherapy for malignant tumors worldwide include camptothecin-based anticancer drugs including irinotecan, a DNA topoisomerase inhibitor, gemcitabine, an antimetabolites, and paclitaxel, a microtubule inhibitor.

[0005] Irinotecan is a semisynthetic and water-soluble analogue of the natural alkaloid camptothecin, also known as CPT-11, which acts to inhibit cell proliferation by preventing DNA replication by inhibiting the dissolution of DNA by topoisomerase. Irinotecan is currently used as a treatment for camptosol. TMIrinotecan is commercially available in aqueous solution formulations such as irinotecan hydrochloride injection. Irinotecan is a prodrug anticancer drug that acts by being converted to the active metabolite SN-38 by carboxylesterase 2 (CES2) in the body. The active form, SN-38, has anticancer activity approximately 100 to 1,000 times higher than irinotecan, but is unstable at the body's pH and is extremely poorly soluble, making it difficult to develop a formulation. In addition, the conversion rate of irinotecan to SN-38 in the body is very low at 2 to 8%, and shows a large variation (more than 4 times) between patients, making it necessary to precisely calculate the dosage (mg / m2) for each patient. 2 ) is essential for the cytotoxic anticancer drug, and there is a problem that its exact effects or side effects cannot be predicted. The active form of SN-38 is an extremely insoluble substance and cannot be dissolved by a general solubilization method, and many solubilization researches and developments are currently underway. Carboxylesterase 2 (CES2), which converts irinotecan into the active form of SN-38, varies greatly in expression ratio in various tumor tissues. CES2 is most abundantly expressed in the small intestine and large intestine, which are the main indications for irinotecan, but is hardly expressed in pancreatic cancer, gallbladder cancer, breast cancer, lung cancer, kidney cancer, prostate cancer, etc. Therefore, if the active form of SN-38 can be directly administered, it is expected that it can exert an anticancer effect even on the tumor tissues where CES2RK is hardly expressed.

[0006] Meanwhile, gemcitabine is a useful anticancer drug, and is currently used as a first-line standard monotherapy treatment, especially for pancreatic cancer. However, its tumor-suppressing efficacy is low, and its effect on extending the survival time of cancer patients is also low. In addition, taxane anticancer drugs such as paclitaxel and nab-paclitaxel (albumin nanoparticle-bound paclitaxel) have limitations in that the sensitivity of tumors varies depending on the patient when used as a monotherapy, and the anticancer effect is not consistent for each patient. Therefore, in order to overcome the limitations of each cytotoxic anticancer drug monotherapy in clinical practice, active research is being conducted to enhance the efficacy and safety of the drug by improving drug delivery technology and combining it with anticancer drugs with different mechanisms of action.

[0007] Throughout this specification, a number of papers and patent documents are referred to and citations are provided, the disclosures of which are incorporated herein by reference in their entirety in order to more clearly describe the level of the technical field to which the present invention pertains and the contents of the present invention.

[0008] Summary of the Invention [Problem to be solved by the invention]

[0009] In general, the low antitumor effect of chemical anticancer drugs is due to the short residence time of the drug in the blood, the low selective drug delivery efficiency to tumor tissue, the high mutation rate of tumors, and the large difference in drug sensitivity between tumors. Therefore, the present inventors have made extensive research efforts to develop a combination therapy that not only efficiently delivers anticancer drugs with high anticancer activity to tumor tissues, but also maximizes antitumor efficacy by combining antitumor drugs with different mechanisms of action to increase tumor sensitivity, and improves safety. As a result, the inventors have confirmed that the combination of an antitumor formulation with a nanoparticle formulation containing hydrophobic camptothecin and hydrophilic camptothecin provides excellent antitumor efficacy, and have completed the present invention.

[0010] It is therefore an object of the present invention to provide a pharmaceutical composition for the treatment of cancer, which is administered in combination with an anti-tumor agent.

[0011] Another object of the present invention is to provide a pharmaceutical combination preparation comprising a particle comprising a hydrophobic camptothecin-based compound, a hydrophilic camptothecin-based compound, and an amphiphilic block copolymer consisting of a hydrophobic block and a hydrophilic block; and an antitumor agent as active ingredients, which can be administered simultaneously, separately or sequentially for the treatment of cancer.

[0012] [Means for solving the problem]

[0013] According to one aspect of the present invention, there is provided a pharmaceutical composition for treating cancer, which is administered in combination with an anti-tumor agent.

[0014] The pharmaceutical composition of the present invention contains as active ingredients particles containing a hydrophobic camptothecin-based compound, a hydrophilic camptothecin-based compound, and an amphiphilic block copolymer consisting of a hydrophobic block and a hydrophilic block.

[0015] In this specification, camptothecin is a topoisomerase inhibitor found in the bark and stem of Camptotheca (Happy tree), and it showed excellent anti-cancer effects in the preclinical stage, but was not used due to its low solubility. Therefore, many researchers are developing camptothecin analogues to increase its solubility, and currently, three camptothecin derivatives, irinotecan, topotecan, and belotecan, have been approved and are used in chemotherapy for cancer.

[0016] As used herein, the term "hydrophobicity" refers to the tendency of non-polar substances to be excluded from water molecules and to aggregate. When a hydrophobic substance is in a hydrophilic liquid, the hydrophobic bonds increase and the hydrophobic substance aggregates as if it were afraid of water.

[0017] In this specification, the term "hydrophilicity" refers to a tendency that is mainly observed in polar substances, and means a property of having a strong affinity for polar solvents such as water and being easily dissolved. For example, the surface of a hydrophilic polymer compound or a micellar colloid of a surfactant is highly hydrophilic.

[0018] In one embodiment of the present invention, the hydrophobic camptothecin-based compound is at least one selected from the group consisting of SN-38 (7-ethyl-10-hydroxycamptothecin), camptothecin, 10-hydroxycamptothecin, and pharma- ceutical acceptable salts thereof, but is not limited thereto.

[0019] In another embodiment of the present invention, the hydrophilic camptothecin-based compound is at least one selected from the group consisting of irinotecan, topotecan, belotecan, exatecan, lurtotecan, sinotecan, rubitecan, 9-nitrocamptothecin, 9-aminocamptothecin, dimatecan, karenitecin, silatecan, diflomotecan, elomotecan, pharma-ceutically acceptable salts thereof, or glucuronide metabolites thereof, and glucuronide metabolites of the hydrophobic camptothecin-based compounds, but is not limited thereto. The hydrophilic camptothecin-based compounds mentioned above may include compounds generally classified as hydrophobic drugs, but the term "hydrophilic" should be understood to mean relative hydrophilicity compared to the above-mentioned hydrophobic camptothecin-based compounds among the components constituting the particles of the present invention. That is, the hydrophilic camptothecin-based compound refers to a camptothecin-based compound that is relatively hydrophilic compared to the hydrophobic camptothecin-based compound contained in the particles of the present invention.

[0020] According to a specific embodiment of the present invention, the hydrophobic camptothecin-based compound constituting the particles of the present invention may be camptothecin, SN-38, or a mixture thereof, and the hydrophilic camptothecin-based compound is irinotecan hydrochloride, topotecan hydrochloride, or a glucuronide analog of SN-38.

[0021] In this specification, the term "copolymer" refers to a polymer made of two or more different monomers. For example, when styrene and acrylonitrile are reacted in the same reaction vessel, a copolymer containing both monomers is formed. A "block copolymer" refers to a copolymer in which a block of one monomer is linked to a block of another monomer. When a block of substance A is followed by a block of substance B, it is expressed as -[-AB-]-. If the chain is simply composed of one of each monomer, it is called AB type, if there is a B block in the middle and an A block at each end, it is called ABA type, and if there are three different blocks in the main chain, it is called ABC type. Block copolymers are mainly formed by ionic polymerization. Unlike other copolymers, this block copolymer has many of the physical properties of a homogeneous polymer made from two monomers.

[0022] In one embodiment of the present invention, the amphiphilic block copolymer constituting the particles of the present invention is composed of an AB or ABA block, where A is a hydrophilic polymer selected from, but not limited to, monomethoxypolyethylene glycol, dimethoxypolyethylene glycol, polyethylene glycol, polypropylene glycol, monomethoxypolypropylene glycol, polyethylene oxide, polyacrylic acid, etc.

[0023] In addition, B is a hydrophobic polymer, and may be polylactic acid, poly-L-lactide, poly-D-lactide, poly-D,L-lactide, poly(lactide-co-glycolide), polyglyconic acid, polyglycolide, polylactic acid-glyconic acid copolymer, polymandelic acid, polycaprolactone, polydioxane-2-one, polyglutamic acid, polyaspartic acid, polyornithine, polyorthoester, derivatives thereof, or one or more compounds selected from these, but is not limited thereto, and it will be apparent to those skilled in the art that any compound capable of forming an amphiphilic block copolymer that can be used in the art may be used.

[0024] In a specific embodiment of the present invention, the amphiphilic block copolymer is mPEG-PDLLA (mPEG-Poly(D,L)Lactic Acid); PEG-PCL [poly(ethylene glycol)-b-poly(carprolactone)]; PEG-PLA [poly(ethylene glycol)-b-poly(lactic acid)]; mPEG-PGA [monomethoxy poly(ethylene glycol)-b-poly(glycolic acid)]; mPEG-PLGA [monomethoxy poly(ethylene glycol)-b-poly(lactide-co-glycolide)]; PEG-PBLA [poly(ethylene glycol)-b-poly(β-benzyl-L-aspartic acid)]; PEG-p(Glu) [poly(ethylene glycol)-b-poly(glutamic acid)]; PEG-p(Asp) [poly(ethylene glycol)-b-poly(aspartic acid)]; and / or PEG-PLA-PEG [poly(ethylene glycol)-b-poly(lactic acid)-b-poly(ethylene glycol)].

[0025] According to the most specific embodiment of the present invention, the characteristics and manufacturing method of the particles of the present invention are described in detail in the applicant's Korean Patent Registration No. 10-2094543, the disclosure of which is incorporated herein by reference.

[0026] The particles containing SN-38 as a hydrophobic camptothecin-based compound, irinotecan hydrochloride as a hydrophilic camptothecin-based compound, and mPEG-PDLLA as an amphiphilic block copolymer, prepared according to the above registered patent or the following examples, were named SNB-101.

[0027] In another embodiment of the present invention, the weight ratio of the hydrophobic camptothecin-based compound to the hydrophilic camptothecin-based compound constituting the particles of the present invention is 1-10:1-10, 1-10:1-5, 1-10:1-3, 1-10:1, 1-5:1-10, 1-3:1-10, 1:1-10, specifically 1-5:1-5, 1-5:1-3, 1-5:1, 1-3:1-5, 1:1-5, more specifically 1-3:1-3, 1-3:1, 1:1-3, but is not limited thereto.

[0028] In a specific embodiment of the present invention, the weight ratio of the hydrophobic camptothecin-based compound to the hydrophilic camptothecin-based compound may be 1:1-10, 1:1-5, 1:1-3, or 1:1-2, and most specifically may be 1:1.59, but is not limited thereto.

[0029] In one embodiment of the present invention, the weight ratio of the sum of the (a) hydrophobic camptothecin-based compound and the hydrophilic camptothecin-based compound to the (b) amphiphilic block copolymer is 1:0.1 to 200, 1:0.5 to 200, 1:1 to 200, 1:2 to 200, 1:5 to 200, 1:10 to 200, 1:50 to 200, 1:100 to 200, 1:150 to 200, 1:0.1 to 100, 1:0.5 to 100, 1:1 1:0.1-50, 1:0.5-50, 1:1-50, 1:5-50, 1:10-50, 1:20-50, 1:0.1-20, 1:0.5-20, 1:1-20, 1:5-20, 1:10-20, 1:0.1-10, 1:0.5-10, or 1:1-10, but is not limited thereto.

[0030] As used herein, the term "~" between two numerical values ​​means an interval between those numerical values, including the numerical values ​​before and after it.

[0031] In one embodiment of the present invention, the above-mentioned particles of the present invention are prepared by the following method. First, hydrophobic camptothecin (e.g., SN-38) is added to an organic solvent together with hydrophilic camptothecin (e.g., irinotecan hydrochloride) and stirred to completely dissolve, and amphiphilic block copolymer (e.g., mPEG-PDLLA) previously dissolved in an organic solvent is added thereto and stirred. This mixture is dried in a rotary evaporator vacuum dryer or vacuum dryer, and an aqueous solvent (e.g., distilled water, PBS) is added to the residue, and ultrasonic waves are applied in an ultrasonic cleaner to prepare the particles of the present invention.

[0032] In one embodiment of the present invention, the organic solvent may be, but is not limited to, C1 to C5 alcohol, (methanol, ethanol, propanol, butanol, n-butanol, iso-propanol, 1-pentanol, 2-butoxyethanol, isobutyl alcohol, etc.), alkyl acetate, acetone, acetonitrile, chloroform, benzene, toluene, xylene, acetone, fluoroalkane, pentane, hexane, 2,2,4-trimethylpentane, decane, cyclohexane, cyclopentane, diisobutylene, 1-pentene, 1-chlorobutane, 1-chloropentane, diisopropyl ether, 2-chloropropane, 1-chloropropane, chlorobenzene, benzene, diethyl ether, diethyl sulfide, dichloromethane, 1,2-dichloroethane, aniline, diethylamine, ether, carbon tetrachloride, THF (Tetrahydrofuran), or a mixture thereof.

[0033] The pharmaceutical composition of the present invention comprises an anti-tumor agent.

[0034] The antitumor agent includes at least one selected from the group consisting of taxane anticancer agents, albumin-bound taxane anticancer agents, vascular endothelial growth factor (VEGF) inhibitors, and gemcitabine.

[0035] In one embodiment of the present invention, the taxane anticancer drug may be at least one selected from the group consisting of paclitaxel, docetaxel, larotaxel, cabazitaxel, and pharma- ceutical acceptable salts thereof, but is not limited thereto.

[0036] In one embodiment of the present invention, the albumin-bound taxane anticancer drug may be at least one selected from the group consisting of albumin-bound paclitaxel (albumin nanoparticle paclitaxel or nab-paclitaxel) and albumin-bound docetaxel (albumin nanoparticle docetaxel or nab-docetaxel), but is not limited thereto.

[0037] In one embodiment of the present invention, the vascular endothelial growth factor (VEGF) inhibitor may be one or more selected from the group consisting of bevacizumab, ranibizumab, and aflibercept, but is not limited thereto.

[0038]

[0039]

[0040]

[0041] In a specific embodiment of the present invention, the hydrophobic camptothecin compound is SN-38, the hydrophilic camptothecin compound is irinotecan hydrochloride, and the amphiphilic block copolymer is mPEG-PDLLA (mPEG-Poly(D,L)Lactic Acid).

[0042] The particles of the present invention prepared according to this specific embodiment have been named SNB-101 by the present inventors.

[0043] The stable nanoparticle formulation of SN-38 and irinotecan hydrochloride (SNB-101) is a formulation in which SN-38, which is extremely poorly soluble, has been solubilized and formulated. SN-38 has excellent anticancer effects, but it has been difficult to utilize due to its low solubility. It can be directly administered, and therefore can exhibit anticancer effects against pancreatic cancer, ovarian cancer, lung cancer, breast cancer, etc., which do not have carboxylesterase, and is expected to be effective in treating new indications.

[0044] The particles as the active ingredient of the pharmaceutical composition of the present invention according to the specific embodiment of the present invention contain 0.1-1 mg / ml, and more preferably 0.2-1.0 mg / ml, of SN-38 based on the particle composition, and 0.1-2 mg / ml, and more preferably 0.2-1.6 mg / ml, of irinotecan hydrochloride based on the particle composition.

[0045] In one embodiment of the present invention, the particles have a double structure, which is composed of an inner structure formed by the hydrophilic camptothecin-based compound and the hydrophobic camptothecin-based compound, and an outer structure formed by the amphiphilic block copolymer.

[0046] In the process of preparing the dual-structure particles of the present invention, first, a hydrophilic camptothecin compound and a hydrophobic camptothecin compound are mixed and dissolved in an organic solvent, and then an amphiphilic polymer (amphiphilic block copolymer) dissolved in an organic solvent is added to the mixture, stirred, and then dried. The dried product is emulsified in an aqueous solvent by ultrasonic treatment, etc., to form water-acidic nanoparticles.

[0047] It is known that an amphiphilic block copolymer, which is formed by combining a hydrophilic block and a hydrophobic block in a specific ratio, self-assembles in an aqueous solution to form water-acidic particles similar to micelles, and therefore it is presumed that the particles of the present invention contain the above-mentioned hydrophilic camptothecin-based compound and hydrophobic camptothecin-based compound inside the water-acidic particles, and the amphiphilic block copolymer forms an external shell. More specifically, in the structure of the water-acidic particles of the present invention, it is presumed that the hydrophobic block of the amphiphilic block copolymer forms a shell toward the internal structure formed by the relatively hydrophobic camptothecin-based compound, and the hydrophilic block forms a shell toward the external aqueous solvent side.

[0048] In one embodiment of the present invention, the dual-structure hydrophilic particles of the present invention are characterized in that they spontaneously form particles when dispersed in an aqueous solution.

[0049] In another embodiment of the present invention, the average diameter of the particles is 2 to 200 nm. When the size of the particles prepared according to the present invention is 200 nm or less, non-selective removal by the reticuloendothelial system (RES) in the body can be avoided, so it is preferable to prepare particles having a uniform particle size of 200 nm or less.

[0050] In one embodiment of the present invention, the water-acidic particles of the present invention can be freeze-dried after mixing with a freeze-drying protection agent such as trehalose or mannitol.

[0051] The average diameter (particle size) of the particles according to the present invention is about 200 nm or less, and the delivery of the drug to the tumor tissue can be maximized by utilizing the enhanced permeation and retention effect (EPR), which is characteristic of tumor tissue. EPR occurs when the normal integrity of the vascular system (especially capillaries) is damaged, causing particles such as nano-sized particle carriers (nanoparticles, liposomes, micelles) to leak from the capillaries and deposit the carriers at the tumor site.

[0052] Therefore, due to the characteristics of the particles (SNB-101) of the present invention, SN-38 and irinotecan contained in SNB-101 can remain in the blood for a long time, and can promote the accumulation of the drug in tumor tissue. Therefore, it is expected that the exposure of the drug to intratumoral cancer cells is increased and the exposure of the drug to normal tissue is decreased, thereby improving safety and increasing the maximum tolerated dose (MTD) of the drug.

[0053] In one embodiment of the present invention, the pharmaceutical composition according to the above aspect of the present invention is for the treatment of cancer. The cancer may be selected from the group consisting of gastric cancer, ovarian cancer, uterine cancer, cervical cancer, small cell lung cancer, non-small cell lung cancer, pancreatic cancer, breast cancer, esophageal cancer, retinoblastoma, oral cancer, salivary gland cancer, laryngeal cancer, pharyngeal cancer, colon cancer, colorectal cancer (rectal cancer), kidney cancer, prostate cancer, melanoma, liver cancer, gallbladder and other biliary tract cancers, thyroid cancer, bladder cancer, brain and central nervous system cancer, bone tumor, skin cancer, non-Hodgkin's and Hodgkin's lymphoma, but is not limited thereto. The brain cancer may be glioma, brain meningioma, neuroendocrine tumor, pituitary tumor, metastatic brain tumor, or skull base tumor. The glioma includes astrocytoma, oligodendroglioma, ependymoma, or a mixture thereof.

[0054] In a specific embodiment of the present invention, the cancer that is the target disease to be treated by the pharmaceutical composition is pancreatic cancer, gastric cancer, breast cancer, lung cancer, colon cancer, or a combination thereof.

[0055] When the particles of the present invention or a composition containing the particles are prepared as a pharmaceutical composition, the pharmaceutical composition of the present invention may contain a pharma- ceutically acceptable carrier. The pharma- ceutically acceptable carrier is one that is generally used in formulation, and includes, but is not limited to, lactose, glucose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil. In addition to the above ingredients, the pharmaceutical composition of the present invention may further include lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives, etc. Suitable pharma- ceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

[0056] In a specific embodiment of the present invention, the pharmaceutical composition of the present invention further comprises an excipient such as sucrose, mannitol, sorbitol, glycerin, trehalose, polyethylene glycol, and a cyclodextrin excipient (alpha, beta, gamma-cyclodextrin, hydroxycyclodextrin, or a derivative of cyclodextrin, etc.), which is added to the particles, which are the active ingredient of the pharmaceutical composition, to act as a cryoprotectant or an osmotic pressure regulator, and is formulated by freeze-drying, solvent evaporation, etc.

[0057] The pharmaceutical composition of the present invention can be administered orally or parenterally. When administered parenterally, it can be administered intravenously, intraarterially, intrarectally, subcutaneously, intravascularly, intramuscularly, intranasally, intramucosally, intradurally, intraperitoneally, intraocularly, or the like, and specifically, it can be administered intravenously.

[0058] The appropriate dosage of the pharmaceutical composition of the present invention varies depending on factors such as formulation method, administration method, age, weight, sex, pathological condition, food, administration time, administration route, excretion rate and reaction sensitivity of the patient, and a doctor of ordinary skill can easily determine and prescribe an effective dosage for the desired treatment or prevention. According to a specific embodiment of the present invention, the daily dosage of the pharmaceutical composition of the present invention is 0.001-100 mg / kg.

[0059] In a specific embodiment of the present invention, the dose and frequency of administration of SN-38 and irinotecan hydrochloride contained in the SNB-101 of the present invention are 1 to 5 mg / m2 per day based on SN-38. 2 For example, the injection can be administered to a human patient (body weight 60 kg; body surface area 1.67 m 2 ) can be administered by intravenous injection at a dose of 1 to 100 mg, specifically 20 mg to 85 mg, more specifically 30 mg to 85 mg, in one to three divided doses per day. However, the dosage and number of administrations are not limited to these amounts.

[0060] The pharmaceutical composition of the present invention may be prepared in the form of a unit dose or in a multi-dose container by formulating it with pharma- ceutically acceptable carriers and / or excipients in a manner that can be easily carried out by a person having ordinary skill in the art to which the present invention pertains. In this case, the dosage form may be in the form of a solution, suspension or emulsion in an oil or aqueous medium, or in the form of an extract, powder, granules, tablets or capsules, and may further include a dispersant or stabilizer.

[0061] The pharmaceutical composition of the present invention can be administered in combination with a known compound or pharmaceutical composition having a therapeutic effect on cancer.

[0062] In one embodiment of the present invention, the known compound or pharmaceutical composition includes at least one selected from the group consisting of taxane anticancer drugs, albumin-bound taxane anticancer drugs, vascular endothelial growth factor (VEGF) inhibitors, and gemcitabine.

[0063]

[0064] According to one aspect of the present invention, there is provided a method for treating cancer, comprising administering to a subject in need thereof (a) a first pharmaceutical composition comprising a hydrophobic camptothecin-based compound, a hydrophilic camptothecin-based compound, and particles comprising an amphiphilic block copolymer consisting of a hydrophobic block and a hydrophilic block, in combination with (b) a second pharmaceutical composition comprising an antitumor agent as an active ingredient or an anticancer therapy.

[0065] In one embodiment of the present invention, the first pharmaceutical composition and the second pharmaceutical composition are administered simultaneously, separately or sequentially.

[0066] In one embodiment of the present invention, the first pharmaceutical composition and the second pharmaceutical composition are administered as a combined formulation or a single formulation.

[0067] In one embodiment of the present invention, the anti-tumor therapy comprises surgery, radiation therapy, or a combination thereof.

[0068] In one embodiment of the present invention, the cancer of the present invention is selected from the group consisting of gastric cancer, ovarian cancer, uterine cancer, cervical cancer, small cell lung cancer, non-small cell lung cancer, pancreatic cancer, breast cancer, esophageal cancer, retinoblastoma, oral cancer, salivary gland cancer, laryngeal cancer, pharyngeal cancer, colon cancer, large intestine cancer (rectal cancer), kidney cancer, prostate cancer, melanoma, liver cancer, gallbladder and other biliary tract cancer, thyroid cancer, bladder cancer, brain cancer, central nervous system cancer, bone tumor, skin cancer, non-Hodgkin's and Hodgkin's lymphoma, but is not limited thereto. The brain cancer may be glioma, brain meningioma, neuroendocrine tumor, pituitary tumor, metastatic brain tumor, or skull base tumor. The glioma includes astrocytoma, oligodendroglioma, ependymoma, or a mixture thereof.

[0069] In specific embodiments of the present invention, the cancer is pancreatic cancer, gastric cancer, breast cancer, lung cancer, colon cancer, or a combination thereof.

[0070]

[0071] According to another aspect of the present invention, there is provided a pharmaceutical combination formulation comprising a particle comprising a hydrophobic camptothecin-based compound, a hydrophilic camptothecin-based compound, and an amphiphilic block copolymer consisting of a hydrophobic block and a hydrophilic block; and an antitumor agent as active ingredients.

[0072] The pharmaceutical combination preparations may be administered with each active ingredient simultaneously, separately or sequentially for the treatment of cancer.

[0073]

[0074] According to yet another aspect of the present invention, there is provided a kit for treating cancer, comprising as a first pharmaceutical composition particles containing a hydrophobic camptothecin-based compound, a hydrophilic camptothecin-based compound, and an amphiphilic block copolymer consisting of a hydrophobic block and a hydrophilic block; and as a second pharmaceutical composition an antitumor agent.

[0075] The first and second pharmaceutical compositions of the kit for treating cancer may be administered simultaneously, separately or sequentially for treating cancer.

[0076] In one embodiment of the present invention, the first pharmaceutical composition and the second pharmaceutical composition are administered as a combined formulation or a single formulation.

[0077] The term "administration" or "administering" as used herein refers to administering a therapeutically effective amount of the composition of the present invention directly to a subject (individual) suffering from cancer, so that the same amount is formed in the subject's body.

[0078] The "therapeutically effective amount" of the composition means the content of the composition sufficient to provide a therapeutic or prophylactic effect to a subject to which the composition is administered, and thus includes a "prophylactically effective amount." In addition, the term "subject" as used herein includes, but is not limited to, humans, mice, rats, guinea pigs, dogs, cats, horses, cows, pigs, monkeys, chimpanzees, baboons, or rhesus monkeys. Specifically, the subject of the present invention is a human.

[0079] The cancer treatment method, pharmaceutical combination preparation, and cancer treatment kit of the present invention contain the same active ingredient as the pharmaceutical composition for cancer treatment, which is one embodiment of the present invention, and use an antitumor agent as a combination preparation, so that the content described with respect to the above-mentioned one embodiment of the present invention applies equally to the overlapping content.

[0080] Effect of the Invention

[0081] The present invention relates to a cancer treatment use of nanoparticles containing poorly soluble camptothecin-based compounds and a combination therapy using the same with a taxane-based antitumor agent. The pharmaceutical composition containing the nanoparticles of the present invention shows a synergistic effect in cancer treatment when used in combination with a taxane-based antitumor agent, and can be easily used as a cancer treatment agent and a combination therapy preparation.

[0082] [Brief description of the drawings]

[0083] [Figure 1] FIG. 1 shows changes in mouse body weight after administration in AsPC-1 subcutaneously transplanted pancreatic cancer model mice.

[0084] [Diagram 2] FIG. 1 shows the tumor-suppressing effect of treatment with a test substance in a mouse model of subcutaneously transplanted AsPC-1 pancreatic cancer, as measured by changes in tumor volume.

[0085] [Diagram 3] FIG. 1 shows the tumor-suppressing effect of test substance treatment in subcutaneously transplanted AsPC-1 pancreatic cancer model mice, as measured by changes in tumor weight.

[0086] [Figure 4] 1 shows photographs of tumors excised on day 22 after treatment with a test substance in a subcutaneously implanted AsPC-1 pancreatic cancer model mouse.

[0087] [Diagram 5] FIG. 1 shows changes in mouse body weight after administration in Hs746T subcutaneously transplanted gastric cancer model mice.

[0088] [Figure 6] FIG. 1 shows the tumor-suppressing effect of treatment with a test substance in a mouse model of subcutaneously transplanted Hs746T gastric cancer, as measured by changes in tumor volume.

[0089] [Figure 7] FIG. 1 is a graph showing the tumor-suppressing effect of treatment with a test substance in a mouse model of subcutaneously transplanted Hs746T gastric cancer, as measured by changes in tumor weight.

[0090] [Figure 8] Photographs of tumors excised from Hs746T subcutaneously transplanted gastric cancer model mice on day 22 after treatment with a test substance.

[0091] [Figure 9]FIG. 1 shows the tumor suppressive effect of treatment with a test substance in a mouse model of subcutaneously transplanted MDA-MB231 breast cancer, as measured by changes in tumor volume.

[0092] [Figure 10] FIG. 1 shows changes in mouse body weight due to treatment with a test substance in a subcutaneously transplanted MDA-MB231 breast cancer model mouse.

[0093] [Figure 11] FIG. 1 is a graph showing the tumor-suppressing effect of treatment with a test substance in A549 subcutaneously transplanted lung cancer model mice, as measured by changes in tumor volume.

[0094] [Figure 12] FIG. 1 shows changes in mouse body weight due to treatment with a test substance in A549 subcutaneously transplanted lung cancer model mice.

[0095] [Figure 13] FIG. 1 shows changes in tumor volume caused by treatment with a test substance in HT-29 subcutaneously transplanted colon cancer model mice.

[0096] [Figure 14] FIG. 1 shows changes in tumor volume caused by treatment with a test substance in a subcutaneously implanted NCI-H69 small cell lung cancer model mouse.

[0097] [Figure 15] FIG. 1 shows photographs of tumors in subcutaneously implanted NCI-H69 small cell lung cancer model mice treated with a test substance.

[0098] [Figure 16] FIG. 1 shows changes in tumor volume due to treatment with a test substance in a subcutaneously transplanted HCT116 colon cancer model mouse.

[0099] [Figure 17] FIG. 1 shows photographs of tumors in subcutaneously transplanted HCT116 colon cancer model mice treated with a test substance.

[0100] [Figure 18] FIG. 1 shows changes in tumor volume caused by treatment with a test substance in A549 subcutaneously implanted non-small cell lung cancer model mice.

[0101] DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0102] The present invention will be described in more detail below with reference to examples. It will be apparent to those skilled in the art that these examples are merely intended to more specifically illustrate the present invention, and that the scope of the present invention is not limited to these examples according to the gist of the present invention.

[0103]

[0104] Working Example

[0105]

[0106] Throughout this specification, "%" used to indicate the concentration of a particular substance is, unless otherwise specified, % (w / w) for solid / solid, % (w / vol) for solid / liquid, and % (vol / vol) for liquid / liquid.

[0107]

[0108] Example 1: Preparation of nanoparticles of the invention (SNB-101)

[0109] The present inventors placed 20 mg of SN-38 (7-ethyl-10-hydroxycamptothecin) as a hydrophobic camptothecin and 30 mg of irinotecan hydrochloride (trihydrate) as a hydrophilic camptothecin in a round-bottom flask, added 10 ml of acetonitrile, and applied ultrasonic waves with a cleaning sonicator to completely dissolve both drugs. Next, 100 mg of methoxypolyethylene glycol-poly(D,L,lactic acid) [mPEG-Poly(D,L)Lactic Acid] (mPEG-PDLLA) was weighed into a round-bottom flask, added 10 ml of an organic solvent (ethanol-acetonitrile 50:50 v / v mixture), and stirred for 30 minutes to completely dissolve. The completely dissolved mPEG-PDLLA solution was gradually added to the drug solution in which SN-38 and irinotecan hydrochloride were dissolved, with stirring for 10 to 15 seconds each time. The mixed solution was mixed in a round-bottom flask, and then dried in a rotary evaporator to completely remove the organic solvent, obtaining a thin film, to which 20 ml of sterile distilled water was added, and ultrasonic waves were applied in an ultrasonic cleaner to completely dissolve the drug film.

[0110] The average particle size of the prepared dual-structure nanoparticles was measured by diluting the sample with distilled water to prepare 1 mg / ml SN-38, and measuring the intensity weight-averaged diameter by dynamic light scattering using a Zetasizer Nano System manufactured by Malvern (UK).

[0111] Next, 200 mg of trehalose and 300 mg of mannitol (D-mannitol) as freeze-drying protective agents were added to 20 ml of sterile distilled water and stirred to completely dissolve.

[0112] Finally, the lyoprotectant solution was added to the mixed drug nanoparticle solution with stirring for 10-15 seconds each time. The final mixed solution was filtered through a 0.22 μm cellulose acetate membrane filter and filled into packaging vials. The filled vials were freeze-dried to obtain the finished product in the form of a dry powder or cake (designated SNB-101).

[0113]

[0114] Example 2: Effect of combined administration of the nanoparticles of the present invention (SNB-101) to pancreatic cancer

[0115]

[0116] 2-1. Experimental materials

[0117] The substances used in the study were 5% glucose injection, irinotecan hydrochloride (Irinotecan HCl; Campto Injection), and TM , CJ Healthcare, Republic of Korea), Nab-paclitaxel (Abraxane) TM The irinotecan hydrochloride (Campto Injection) was used. TM ) was purchased from CJ Healthcare at 100 mg / 5 mL. Abraxane was purchased from Abraxis BioScience, LLC (USA) and used as a 100 mg / vial lyophilized injection.

[0118] The nanoparticle composition (SNB-101) containing SN-38 and irinotecan hydrochloride used in the present invention is a stable nanoparticle formulation as an injection, and its manufacturing method is introduced in Korean Patent Registration No. 10-2094543. The intravenous injection of SNB-101 contains 10 mg of the main ingredient per vial of SN-38 and 15.9 mg of irinotecan hydrochloride, and can be prepared by diluting with 9.64 mL of 5% glucose injection USP and injecting over a period of 90 minutes.

[0119]

[0120] 2-2. Test method

[0121] 1) AsPC-1 cell culture

[0122] AsPC-1 was used as a pancreatic cancer cell line for the xenograft model of pancreatic cancer. The AsPC-1 cell line is an aggressive cell line known to be resistant to gemcitabine. AsPC-1 cells were cultured at 175 cm in Roswell Park Memorial Institute medium (RPMI-1640) containing 10% FBS (fetal bovine serum) and 1% penicillin / streptomycin. 2 FBS, RPMI-1640, and penicillin / streptomycin were purchased from ATCC (Manassas, VA) and used in the experiments. The cells were maintained at 37°C and 5% CO 2 The cells were cultured in an incubator. The cultured cancer cells were refeeded 2-3 times a week, washed with phosphate buffered saline (PBS, pH 7.4), and attached cells were separated with 0.05% trypsin-0.02% EDTA. The separated cells were centrifuged (3 minutes, 1500 rpm) to accumulate cancer cells, and the medium was added to the accumulated cancer cells, mixed well with a pipette to disperse them evenly, and then subcultured for use in the experiment.

[0123]

[0124] 2) Induction of pancreatic cancer xenograft mouse animal model

[0125] The animals used in this study were BALB / c nu / nu male mice (approximately 5 weeks old, average body weight 19g ± 20%), which are widely used in xenograft animal models, and were allowed to undergo an acclimation process for approximately 5 to 7 days after procurement. On the day of inoculation, the cultured cancer cells were washed with phosphate buffered saline (PBS, pH 7.4), and then the attached cells were separated using 0.05% trypsin-0.02% EDTA. The separated cells were centrifuged (3 minutes, 1500 rpm), diluted with PBS, and then diluted to 5 × 10 per mouse. 6 The tumor cells were subcutaneously injected into the right flank of each mouse at a concentration of 0.2 cells / mL, and it was confirmed that the tumor cell suspension did not flow out from the injection site.

[0126] Group composition and dosage setting

[0127] Mice were divided into groups with an average pancreatic cancer tumor volume of 200±20 mm 3 The composition and dosage of the control and test groups were as follows:

[0128]

[0129] [Table 1]

[0130] * Dose notation of SNB-101 as SN-38 / irinotecan hydrochloride

[0131]

[0132] 3) Administration site and method

[0133] The test substance was administered intravenously via the tail vein (iv) at an individual dose calculated based on the body weight immediately before administration. The test substance was administered once a week (on days 0, 7, 14, 17, and 21) for a total of five times, and the mice were necropsied on the 22nd.

[0134]

[0135] 4) Observation and inspection items

[0136] During the test period, clinical symptoms were observed once a day, and tumor volumes were measured three times a week along with drug administration. After the test was completed, the animals were anesthetized with isoflurane, bled, and tumor tissues were individually excised. The excised tumors were weighed and photographed. The tumor volume was calculated by measuring the long and short axis sizes of the tumor using a vernier caliper (Mitutoyo Corp., Model No.: CD-15CPX, Japan) and the tumor volume was calculated according to the following formula 1.

[0137] formula 1

[0138] Tumor volume = (short axis) 2 × Long axis × 0.5

[0139]

[0140] 5) Statistical analysis

[0141] The test results were statistically analyzed by Mann-Whitney analysis using a commercial statistical program (IBM SPSS statistics version 19.0).

[0142]

[0143] 2-3.Test results

[0144] 1) Weight measurement results

[0145] The body weight was measured on the 22nd day after administration of the test substance (the day of necropsy). The results are shown in Table 2 and FIG.

[0146] [Table 2]

[0147] Compared to G1 (vehicle group), the weight loss rates in all test substance administration groups (G3 to G8) were 10.0%, 10.0%, 10.0%, 15.4%, 12.4%, and 17.4%, respectively, and it was determined that the weight loss rate did not exceed 20% and had not reached the critical minimum weight (Figure 1).

[0148]

[0149] 2) Tumor volume measurement results

[0150] The pancreatic cancer tumor volume measurement results are shown in Table 3 below and FIG.

[0151]

[0152] [Table 3]

[0153] The volume of pancreatic cancer tumors continued to increase in the vehicle group (G1) and the nab-paclitaxel single-administration group (G2; 5 mg / kg) until 22 days after group separation, and the tumor growth rate was higher than that of the other groups.

[0154] Compared to the vehicle group, the tumor volume was significantly reduced in the nab-paclitaxel + SNB-101 (high dose) combination treatment group (G8) from day 7, and the SNB-101 (low and high dose) alone treatment groups (G5, G6) and the nab-paclitaxel + SNB-101 (high dose) combination treatment group (G8) all showed significant reductions compared to the vehicle group from day 17 to day 22 (p<0.05).

[0155] On day 22, the nab-paclitaxel + irinotecan hydrochloride combination group (G4) showed a significant decrease of 32.4% compared to the nab-paclitaxel alone group (G2) and a 12.8% decrease compared to the irinotecan alone group (G3), but no statistically significant difference was observed (p>0.05).

[0156] In a comparison between the groups administered SNB-101 (low and high doses) alone (G5 vs. G6), the high dose alone group (G6) showed a significant decrease of 11.0% compared to the low dose alone group (G5), indicating that the pancreatic cancer tumor suppression effect of SNB-101 was dose-dependent (10 / 15.9 mg / kg vs. 20 / 31.8 mg / kg as SN-38 / Irinotecan HCl).

[0157] On the other hand, when comparing the nab-paclitaxel alone (G2) or SNB-101 (low and high dose) alone (G5, G6) with the nab-paclitaxel + SNB-101 (low and high dose) combined administration groups (G7, G8), both combined administration groups showed higher tumor growth inhibition effects than either alone administration group (p<0.05, <0.01). When comparing the nab-paclitaxel + SNB-101 (low and high dose) groups (G7, G8), the tumor growth inhibition effect increased statistically significantly with increasing SNB-101 dose (p<0.05).

[0158] On the other hand, no tumor suppression effect was observed with nab-paclitaxel (5mg / kg) alone (G2), but the nab-paclitaxel + high-dose SNB-101 group (G8) (20 / 31.9mg / kg as SN-38 / irinotecan HCl) showed a higher tumor growth suppression effect (52.6%) than the tumor suppression effect (38.4%) of the high-dose SNB-101 alone group (G6). In addition, the nab-paclitaxel + high-dose SNB-101 group (G8) showed a statistically significant superior tumor growth suppression effect (29.5%) when compared with the nab-paclitaxel + irinotecan hydrochloride combination group (G4) (p<0.05).

[0159] These results indicate that combined administration of nab-paclitaxel and SNB-101 may exert a synergistic effect in suppressing pancreatic tumor growth.

[0160]

[0161] 3) Tumor weight measurement results

[0162] The results of measuring the tumor weight are shown in Table 4 below and FIG.

[0163] [Table 4]

[0164]

[0165] As shown in Table 4 and Figure 3 above, the tumor weight of the non-treated group (vehicle group, G1) was 0.62 ± 0.11 g on average, the tumor weight of the nab-paclitaxel treated group (G2) was 0.64 ± 0.15 g, and the tumor weight of the SNB-101 (low and high dose) single administration group (G5, G6) was 0.47 ± 0.10 g and 0.38 ± 0.11 g, respectively. In addition, the tumor weight of the nab-paclitaxel + SNB-101 (low and high dose) combined administration group (G7, G8) was 0.43 ± 0.10 and 0.31 ± 0.05 g, respectively, and the combined administration group showed a tumor inhibition effect of 50% or more compared to the nab-paclitaxel single administration group (G2) and 10 to 22.5% or more compared to the SNB-101 (low and high dose) single administration group (G5, G6). From the above results, it was confirmed that nab-paclitaxel and SNB-101 have the potential to be a highly effective combination therapy in the treatment of pancreatic cancer.

[0166] The nab-paclitaxel + irinotecan hydrochloride combination group (G4) showed a significant decrease of 30.8% compared to the nab-paclitaxel alone group (G2), and when comparing groups receiving SNB-101 (low and high doses) alone, the high dose group (G6) showed a 20.0% decrease compared to the low dose group (G5).

[0167]

[0168] Example 3: Effect of combined administration of the nanoparticles of the present invention (SNB-101) to gastric cancer

[0169]

[0170] 3-1. Experimental materials

[0171] The substances used in the study were 5% glucose injection, irinotecan hydrochloride (Irinotecan HCl; Campto Injection), and TM , CJ Healthcare, Republic of Korea), Docetaxel (Taxotere Injection) TM (Sanofi Aventis Korea) and a nanoparticle composition containing SN-38 and irinotecan hydrochloride (designated SNB-101) were used. TM ) was 100mg / 5mL and was purchased from CJ Healthcare. Docetaxel was purchased from Sanofi Aventis Korea and was 20mg / mL. The nanoparticle composition (SNB-101) containing SN-38 and irinotecan hydrochloride used in the present invention is a stable nanoparticle formulation as an injection, and its manufacturing method is introduced in Korean Patent Registration No. 10-2094543. The intravenous injection of SNB-101 contains 10mg of SN-38 and 15.9mg of irinotecan hydrochloride per vial, and can be prepared by diluting with 9.64mL 5% glucose injection USP and injecting for 90 minutes.

[0172]

[0173] 3-2. Test method

[0174] 1) Culture of gastric cancer cell line Hs746T

[0175] Hs746T was used as a gastric cancer cell line for the gastric cancer xenograft model. Hs746T cells were cultured at 175 cm in DMEM (Dulbecco's modified eagle's medium) containing 10% FBS (fetal bovine serum) and 1% penicillin / streptomycin. 2 FBS, DMEM, and penicillin / streptomycin were purchased from ATCC (Manassas, VA) and used in the experiments. The cells were maintained at 37°C in 95% air and 5% CO. 2The cultured cancer cells were re-fed 2-3 times a week, washed with phosphate buffered saline (PBS, pH 7.4), and then attached cells were separated with 0.05% trypsin-0.02% EDTA. The separated cells were centrifuged (3 min, 1500 rpm) to collect the cancer cells, which were then added to the medium and mixed well with a pipette to disperse them evenly. The cells were then used for the experiments after subculture.

[0176] On the day of inoculation, the cultured cancer cells were washed with PBS, and the attached cells were separated with 0.05% trypsin-0.02% EDTA. The separated cells were centrifuged (3 min, 1500 rpm), diluted with PBS, and then diluted to 4 × 10 per mouse. 6 The concentration was adjusted to cells / 0.2 mL.

[0177]

[0178] 2) Induction of gastric cancer xenograft mouse model

[0179] The animals used in this study were BALB / c nu / nu male mice (approximately 5 weeks old, average body weight 19g ± 20%), which are widely used in xenograft animal models, and were allowed to undergo an acclimation process for approximately 5 to 7 days after acquisition. On the day of inoculation, the cultured cancer cells were washed with phosphate buffered saline (PBS, pH 7.4), and the attached cells were separated using 0.05% trypsin-0.02% EDTA. The separated cells were centrifuged (3 minutes, 1500 rpm), diluted with PBS, and then diluted to 5 × 10 per mouse. 6 The tumor cells were subcutaneously injected into the right flank of each mouse at a concentration of 0.2 cells / mL, and it was confirmed that the tumor cell suspension did not flow out from the injection site.

[0180]

[0181] Group composition and dosage setting

[0182] The mice were divided into groups with gastric cancer tumor volumes of 80±20 mm 3The composition and dosage of the control and test groups were as follows:

[0183] [Table 5]

[0184] * Dose notation of SNB-101 as SN-38 / irinotecan hydrochloride

[0185]

[0186] 3) Administration site and method

[0187] The test substance was administered intravenously via the tail vein (iv) at an individual dose calculated based on the body weight immediately before administration. The test substance was administered once a week (on days 0, 7, and 14) for a total of three times, and the mice were necropsied on the 15th.

[0188]

[0189] 4) Observation and inspection items

[0190] During the test period, clinical symptoms were observed once a day, and tumor volumes were measured three times a week along with drug administration. After the test was completed, the animals were anesthetized with isoflurane, bled, and tumor tissues were removed from each animal. The removed tumors were weighed and photographed. The tumor volume was calculated by measuring the long and short axis sizes of the tumor using a vernier caliper (Mitutoyo Corp., Model No.: CD-15CPX, Japan) and using the following formula 1:

[0191] formula 1

[0192] Tumor volume = (short axis) 2 × Long axis × 0.5

[0193]

[0194] 5) Statistical analysis

[0195] The test results were statistically analyzed by Mann-Whitney analysis using a commercial statistical program (IBM SPSS statistics version 19.0).

[0196]

[0197] 3-3.Test results

[0198] 1) Weight measurement results

[0199] The body weight (g) was measured on the 15th day after administration of the test substance (the day of necropsy). The results are shown in Table 6 and FIG.

[0200] [Table 6]

[0201] Unit:g,Data are expressed in mean ± SD(n=9)

[0202] G1: Vehicle control (5% glucose injection)

[0203] G2:Positive control 1(5mg / kg / day Docetaxel)

[0204] G3:Positive control 2(60mg / kg / day Irinotecan HCl)

[0205] G4:Positive control 3(5mg / kg / day Docetaxel + 60mg / kg / day Irinotecan HCl)

[0206] G5:Test article1(20 / 31.8mg / kg / day(as SN-38 / Irinotecan HCl)SNB-101)

[0207] G6:Test article 2(5mg / kg / day Docetaxel + 20 / 31.8mg / kg / day(as SN-38 / Irinotecan HCl)SNB-101)

[0208]

[0209] No significant difference in body weight was observed in any of the groups at the time of acquisition or group separation. As a result of measuring body weight on the 15th day after administration of the test substance (the day of necropsy), weight loss was observed in the combination administration groups G4 (docetaxel + irinotecan HCl) and G6 (docetaxel + SNB-101), and the weight of the combination administration group on the 15th day was significantly decreased compared to the vehicle control groups G1 (5% glucose injection), G2 (docetaxel), G3 (irinotecan HCl), and G4 (SNB-101) alone. Compared to G1 (vehicle group), the weight loss rate did not exceed 20% in all test substance administration groups (G2 to G6), and it was judged that the critical minimum weight had not been reached (Figure 5).

[0210]

[0211] 2) Tumor volume measurement results

[0212] Gastric cancer tumor volume (mm 3 The measurement results are shown in Table 7 below and FIG.

[0213] [Table 7]

[0214] Unit:g,Data are expressed in mean ± SD(n=9)

[0215] *Compared with G1:*p<0.05,**p<0.01; # Compared with G2: # p<0.05, ## p < 0.01; ¶ Compared with G3: ¶ p<0.05,¶¶ p<0.01

[0216] G1: Vehicle control (5% glucose injection)

[0217] G2:Positive control 1(5mg / kg / day Docetaxel)

[0218] G3:Positive control 2(60mg / kg / day Irinotecan HCl)

[0219] G4:Positive control 3(5mg / kg / day Docetaxel + 60mg / kg / day Irinotecan HCl)

[0220] G5:Test article 1(20 / 31.8mg / kg / day(as SN-38 / Irinotecan HCl)SNB-101)

[0221] G6:Test article 2(5mg / kg / day Docetaxel + 20 / 31.8mg / kg / day(as SN-38 / Irinotecan HCl)SNB-101)

[0222]

[0223] The gastric cancer tumor volume measurement showed that it continued to increase in the vehicle control group G1 up to 15 days after group separation, and the tumor growth rate was higher than that of the other groups.

[0224] Compared to the vehicle control group G1, the tumor volume was significantly reduced from day 7 in G4 (docetaxel + irinotecan HCl combination administration group), G5 (SNB-101 alone administration group) and G6 (docetaxel + SNB-101 combination administration group), and by day 15, it was significantly reduced in all of the positive control substance (G2, G3, G4) and test substance administration groups (G5, G6).

[0225] G4 (docetaxel + irinotecan HCl combination group) showed a statistically significant decrease from the 11th day compared to G2 (docetaxel alone group), and G6 (docetaxel + SNB-101 combination group) showed a significant decrease from the 9th day compared to G2 (docetaxel alone group) and G3 (irinotecan HCl alone group).

[0226]

[0227] 3) Tumor weight measurement results

[0228] The results of measuring the tumor weight are shown in Table 8 below and FIG.

[0229] [Table 8]

[0230] Data are expressed in mean ± SD(n=9);

[0231] *Compared with G1: ** p < 0.01;

[0232] # Compared with G2: # p<0.05, ## p < 0.01;

[0233] ¶ Compared with G3: ¶ p<0.05

[0234] G1: Vehicle control (5% glucose injection)

[0235] G2:Positive control 1(5mg / kg / day Docetaxel)

[0236] G3:Positive control 2(60mg / kg / day Irinotecan HCl)

[0237] G4:Positive control 3(5mg / kg / day Docetaxel + 60mg / kg / day Irinotecan HCl)

[0238] G5:Test article1(20 / 31.8mg / kg / day(as SN-38 / Irinotecan HCl)SNB-101)

[0239] G6:Test article 2(5mg / kg / day Docetaxel + 20 / 31.8mg / kg / day(as SN-38 / Irinotecan HCl)SNB-101)

[0240]

[0241] As shown in Table 8 and Figure 7 above, tumors were excised and compared for tumor weight. As a result, the tumor weight was significantly reduced in the positive control substance (G2, G3, G4), the test substance alone (G5), and the combined administration group (G6) compared to the vehicle control group G1. Compared to G2 (docetaxel alone administration group), G4 (docetaxel + irinotecan HCl combined administration group), G5 (SNB-101 alone administration group), and G6 (docetaxel + SNB-101 combined administration group) showed a significant reduction. In addition, compared to the irinotecan HCl alone administration group, the SNB-101 alone administration group and the docetaxel + SNB-101 combined administration group showed a significant reduction.

[0242] Therefore, under the test conditions, the positive control substances G2 (5mg / kg / day Docetaxel), G3 (60mg / kg / day Irinotecan HCl alone), G4 (5mg / kg / day Docetaxel + 60mg / kg / day Irinotecan HCl in combination), the test substances G5 (20 / 31.8mg / kg / day (as SN-38 / Irinotecan HCl) SNB-101 alone) and G6 (5mg / kg / day Docetaxel + 20 / 31.8mg / kg / day (as SN-38 / Irinotecan HCl) SNB-101 in combination) were determined to be effective in inhibiting tumor growth in the xenograft model using the gastric cancer cell line Hs746T, and the combined administrations (G4, G6) showed higher tumor inhibition than the positive control substances alone (G2, G3).

[0243]

[0244] Example 4: Effect of combined administration of the nanoparticles of the present invention (SNB-101) against breast cancer and lung cancer

[0245]

[0246] 4-1. Experimental materials

[0247] The substances used in the study were 5% glucose injection, docetaxel (docetaxel; Taxotere injection), TM , Sanofi Aventis Korea), and a nanoparticle composition containing SN-38 and irinotecan hydrochloride (designated SNB-101). Docetaxel was purchased from Sanofi Aventis Korea and was 20 mg / mL.

[0248] The nanoparticle composition (SNB-101) containing SN-38 and irinotecan hydrochloride used in the present invention is a stable nanoparticle formulation as an injection, and its manufacturing method is introduced in Korean Patent Registration No. 10-2094543. The intravenous injection of SNB-101 contains 10 mg of SN-38 and 15.9 mg of irinotecan hydrochloride per vial, and can be diluted with 9.64 mL 5% glucose injection USP and infused over a period of 90 minutes.

[0249]

[0250] Test method

[0251] 1) Cultivation of breast cancer cell lines (MDA-MB-231) and lung cancer cell lines (A549)

[0252] MDA-MB-231 and A549 were used as breast cancer and lung cancer cell lines for the xenograft model, respectively. Frozen cell lines MDA-MB-231 and A549 were thawed and cultured in RPMI 1640 medium-10% FBS-1% penicillin / streptomycin and RPMI 1640 medium-5% FBS-1% penicillin / streptomycin at 37°C and 5% CO. 2 The thawed cells were cultured for over a week and subcultured every 2-3 days to check for viability (trypan blue), bacterial and yeast infection (cell images), and mycoplasma infection (MycoAlert Mycoplasma detection kit), and cells with a viability of 90% or more when stained with trypan blue were used for transplantation.

[0253]

[0254] 2) Induction of breast and lung cancer xenograft mouse models

[0255] The animals used in this study were BALB / c nu / nu male mice (approximately 5 weeks old, average body weight 19g ± 20%), which are widely used in xenograft animal models, and were allowed to undergo an acclimation process for approximately 5 to 7 days after acquisition. On the day of inoculation, the cultured cancer cells were washed with phosphate buffered saline (PBS, pH 7.4), and the attached cells were separated using 0.05% trypsin-0.02% EDTA. The separated cells were centrifuged (3 minutes, 1500 rpm) and diluted with PBS, and then 5 × 10 MDA-MB-231 and A549 cells were each injected into each mouse. 6 The tumor cells were subcutaneously implanted into the right shoulder of Balb / c-nu mice at a dose of 100 μl / cell, and it was confirmed that the tumor cell suspension did not flow out from the injection site.

[0256]

[0257] 3) Group composition, administration dose, administration site and administration method

[0258] Mice were divided into groups with an average tumor volume of 90-105 mm 3 The groups were separated by paired-matching at that time, and the composition and administration doses of the control and test groups are shown in Tables 9 and 10.

[0259] [Table 9]

[0260] * The docetaxel concentration was changed from 5 mg / kg (1st to 3rd doses) to 15 mg / kg (4th to 6th doses) during the study.

[0261] ** Drug dosing frequency for all groups was changed during the course of the study from once weekly for 3 weeks (Q1W x 3 times) to once weekly for 6 weeks (Q1W x 6 times).

[0262] *** The dose of 20 mg / kg of SNB-101 is based on the content of SN-38.

[0263]

[0264] [Table 10]

[0265] * The docetaxel concentration was changed from 5 mg / kg (first dose) to 10 mg / kg (second to fifth doses) during the course of the study.

[0266] ** Drug dosing frequency for all groups was changed during the course of the study from once weekly for 3 weeks (Q1W x 3 times) to once weekly for 5 weeks (Q1W x 5 times).

[0267] *** The dose of 20 mg / kg of SNB-101 is based on the content of SN-38.

[0268]

[0269] Tumor volume and body weight were measured twice a week (every 3 or 4 days) from the time of classification until the end of the study, and a tumor growth curve was created based on the tumor volume from the start of drug administration to the end of the study.

[0270] When severe necrosis and weight loss were found, the animals were euthanized in accordance with the Institutional Animal Care and Use Ethics Committee (IACUC) regulations and in consultation with the client, and the study was terminated. The tumors were then excised and photographed.

[0271]

[0272] 4-3. Results of breast cancer xenograft model test using MDA-MB-231

[0273] 1) Tumor volume measurement and result analysis

[0274] On the final day of the experiment (35 days after drug administration), the tumor growth inhibitory efficacy of G2 (docetaxel alone), G3 (SNB-101 alone), and G4 (SNB-101 + docetaxel combination administration) groups was confirmed compared to G1 (vehicle).

[0275] The mean tumor volumes in the G1 (vehicle), G2 (docetaxel), G3 (SNB-101), and G4 (SNB-101 + docetaxel) groups were 1,662.48 mm 3 , 1,228.93mm 3 , 657.30mm 3 , 364.10mm 3 The combined administration (G4) was more effective in suppressing tumor formation than the single administrations (G2 and G3). The results are shown in FIG.

[0276]

[0277] On the final day of the experiment, tumor growth inhibition (TGI) relative to the vehicle was calculated using the following formula:

[0278] formula 2

[0279] Tumor growth inhibition rate (TGI) (%)={1-(△T / △C)}×100

[0280] The TGI by group is shown in Table 11.

[0281] [Table 11]

[0282]

[0283] As shown in Table 11, the TGI compared to vehicle was 27.6% in the group administered with docetaxel alone (G2), 64.0% in the group administered with SNB-101 alone (G3), and 82.6% in the group administered with SNB-101 + docetaxel in combination (G4). From the above results, it was confirmed that the combined administration of SNB-101 of the present invention and docetaxel has a synergistic effect on breast cancer cells.

[0284]

[0285] 2) Weight measurement results

[0286] The weight measurement results by group are shown in FIG.

[0287] At the start of drug administration (day 0), the body weights of the vehicle, docetaxel, SNB-101, and SNB-101 + docetaxel groups were 19.70g, 19.63g, 20.99g, and 21.46g, respectively, and at the end of the study (day 35), they were 22.46g, 20.25g, 22.43g, and 19.37g. In other words, it was confirmed that the weight loss rate did not exceed 20% and did not reach the critical minimum weight.

[0288]

[0289] 3) Conclusion

[0290] In summary, in a breast cancer cell line MDA-MB-231 xenograft model, the combination of SNB-101 and docetaxel (G4) was found to have a significantly superior tumor growth inhibitory effect compared to docetaxel (G2) and SNB-101 alone (G3).

[0291]

[0292] 4-4. Results of the A549 lung cancer xenograft model test

[0293] 1) Tumor volume measurement and result analysis

[0294] On the final day of the experiment (37 days after drug administration), the tumor growth inhibitory efficacy of G2 (docetaxel alone), G3 (SNB-101 alone), and G4 (SNB-101 + docetaxel combined administration) groups was confirmed compared to G1 (vehicle).

[0295] The mean tumor volumes in the G1 (Vehicle), G2 (Docetaxel), G3 (SNB-101), and G4 (SNB-101 + Docetaxel) groups were 704.10 mm 3 , 402.70mm 3 , 557.60mm 3 , 181.08mm 3 The combined administration (G4) was more effective in suppressing tumor formation than the single administrations (G2 and G3). The results are shown in FIG.

[0296]

[0297] On the final day of the experiment, tumor growth inhibition (TGI) relative to the control vehicle was calculated using the following formula.

[0298] formula 2

[0299] Tumor growth inhibition rate (TGI) (%)={1-(△T / △C)}×100

[0300] The TGI by group is shown in Table 12.

[0301] [Table 12]

[0302]

[0303] As shown in Table 12, the TGI compared to vehicle was 50.0% in the docetaxel alone administration group (G2), 24.3% in the SNB-101 alone administration group (G3), and 86.7% in the SNB-101 + docetaxel combination administration group (G4).

[0304] From the above results, it was confirmed that there was a synergistic effect in the combined administration of SNB-101 of the present invention and docetaxel on the lung cancer cell line A549 xenograft model.

[0305]

[0306] 2) Weight measurement results

[0307] The results of weight measurement by group are shown in FIG.

[0308] At the start of drug administration (day 0), the body weights of the vehicle, docetaxel, SNB-101, and SNB-101 + docetaxel groups were 17.86g, 17.93g, 18.04g, and 18.68g, respectively, and at the end of the study (day 37), they were 20.19g, 19.10g, 19.79g, and 17.97g. In other words, it was confirmed that the weight loss rate did not exceed 20% and did not reach the critical minimum weight.

[0309]

[0310] 3) Conclusion

[0311] In summary, in the lung cancer cell line A549 xenograft model, the combination of SNB-101 and docetaxel (G4) showed a significantly superior tumor growth inhibitory effect compared to docetaxel (G2) and SNB-101 alone (G3).

[0312]

[0313] Example 5: Evaluation of synergistic effect of the nanoparticles of the present invention (SNB-101) in combination with bevacizumab (trade name: Avastin) in the treatment of colon cancer

[0314] Experiments were conducted to evaluate therapeutic synergy in colon cancer when combined with bevacizumab, a monoclonal antibody and targeted anticancer drug.

[0315]

[0316] 5-1.Test method

[0317] 1) Experimental animals

[0318] A total of 150 BALB / c nude mice (5-week-old males) were obtained, and after approximately one week of acclimation, HT-29 colon cancer cells were transplanted subcutaneously into the right buttock. Seven days after tumor cell transplantation, 50 transplanted mice were again selected based on tumor volume, and 10 mice per group were used in this experiment.

[0319]

[0320] 2) Tumor cell transplantation

[0321] HT-29 (KCLB No. 300038, Korea Cell Line Bank) human colon cancer cells were cultured in RPMI 1640 medium supplemented with 10% FBS (fetal bovine serum) and 100 IU / ml penicillin / streptomycin at 37°C and 5% CO 2 The cells were maintained by subculture in an incubator at 3.0 × 10 6 A tumor cell suspension was prepared at 0.1 cells / ml, and 0.1 ml of the HT-29 tumor cell suspension was implanted subcutaneously in the right hip of each mouse to form a solid tumor mass.

[0322]

[0323] 3) Group separation

[0324] Eight days after transplantation of the HT-29 colon cancer cell line, mice were selected based on tumor volume and randomly divided into a total of five groups (10 mice / group). Drug administration groups were set as shown in Table 13 below.

[0325] [Table 13]

[0326] *:dose as SN-38 / Irinotecan HCl

[0327]

[0328] 4) Drug Administration

[0329] The test substances, SNB-101 and bevacizumab (5mg / kg), were administered alone and in combination a total of three times on the same day (days 1, 4, and 7). SNB-101 was administered intravenously after calculating the individual dose based on the body weight at the time of administration, and bevacizumab was administered intraperitoneally. Necropsy was performed on the 28th. In the control group, 5% dextrose solution used as a vehicle was administered in the same manner at a dose of 10mL / kg.

[0330]

[0331] 5-2.Test results

[0332] The test results are shown in Table 14 below as mean ± SD.

[0333] [Table 14]

[0334]

[0335] As shown in Table 14 above, compared with the SNB-101 alone group (G2) and the bevacizumab alone group (G3), the combination group (G5) demonstrated a highly superior therapeutic synergy effect in the treatment of colon cancer. The tumor growth graph corresponding to the data in Table 14 above is shown in Figure 13. The statistical significance of the tumor volume difference at different times between groups was analyzed by Two-way ANOVA with Holm-Sidak multiple comparison test (*, P<0.05; **, P<0.01; ***, P<0.001, ns: non-significant).

[0336]

[0337] Example 6: Evaluation of synergistic effect of the nanoparticles of the present invention (SNB-101), nab-paclitaxel and gemcitabine in the treatment of pancreatic cancer

[0338]

[0339] 6-1. Experimental materials

[0340] - BALB / c-nu mouse (Orient Bio-Cabinet Center), male, 6 weeks old

[0341] - Pancreatic cancer AsPC-1 cell line (ATCC, Cat.No.CRL-1682)

[0342] - Test drug: SNB-101 (SN-38 10 mg, Irinotecan HCl 3H 2 O 15.9mg / vial), nab-paclitaxel (Celgene Korea, Abraxane, Lot.No.6200676A), gemcitabine (Lilly Korea, Lot.No.186053A)

[0343] - Vehicle: 5% glucose injection 50ml (CJ Healthcare, insurance code 640001361)

[0344] - RPMI1640 medium(Gibco,A10491-01) supplemented with 10% fetal bovine serum(Gibco,Cat.No.16000)

[0345] - 1% Penicillin / Streptomycin (Gibco, Cat. No. 15140122)

[0346] - Trypsin-EDTA(Gibco,Cat.No.25200-072)

[0347] - Cold-DPBS(Hyclone,Cat.No.SH30258.02)

[0348] - Trypan blue(Gibco,Cat.No.15250061)

[0349] - MycoAlert Mycoplasma detection kit(LONZA,Cat.No.LT-07-318)

[0350] - 150mm cell culture dish, Conical tubes, Pipettes

[0351] - Mouse ear tags, syringes, calipers

[0352]

[0353] 6-2.Test method

[0354] Pancreatic cancer cell line AsPC-1, provided by the National Center for Digestive Disease T2B Infrastructure Development (NCEED), was cultured in RPMI1640 medium, 10% FBS, 1% penicillin / streptomycin at 37°C and 5% CO 2 The thawed cells were cultured for over a week, subcultured at 2-3 day intervals, and checked for viability, bacterial and yeast infection, and mycoplasma infection (MycoAlert Mycoplasma detection kit). Cells with a viability of 90% or more were stained with trypan blue and used for transplantation. The cells were suspended in cold PBS after floating with trypsin-EDTA. 5 × 10 AsPC-1 cells were injected into the right thigh of a BALB / c-nu mouse. 6 The tumor formation and growth were periodically observed, and the tumor length was measured to calculate the tumor volume according to the following formula:

[0355] [Tumor volume = (a 2 b) / 2, a is the minor axis and b is the major axis]

[0356] The average tumor volume of all mice on the day of administration was 123.6 mm 3 The mice were grouped by pair matching when the animals were in the same group. Each group was administered a drug according to the dosage regimen shown in Table 15 below, and observed for two weeks.

[0357] [Table 15]

[0358] *:dose as SN-38 / Irinotecan HCl

[0359]

[0360] Tumor volume and body weight were measured twice a week (every 3 or 4 days) from the time of classification until the end of the study, and the degree of tumor growth inhibition from the start of drug administration until the end of the study was evaluated.

[0361]

[0362] 6-3.Test results

[0363] At the end of the experiment, tumor growth inhibition (TGI) relative to the vehicle was calculated using the above-mentioned formula 2.

[0364] The TGI by group is shown in Table 16.

[0365] [Table 16]

[0366] As shown in Table 16 above, the TGI compared to vehicle was 72.5% in the SNB-101 alone group (G2) and 56.3% in the nab-paclitaxel + gemcitabine group (G3). In contrast, the TGI was 83.4% in the SNB-101 + nab-paclitaxel + gemcitabine combination group (G4), confirming that the therapeutic synergistic effect of the combination of SNB-101 + nab-paclitaxel + gemcitabine during pancreatic cancer treatment is very excellent.

[0367] Furthermore, no macroscopic side effects, weight loss, or toxicity in blood analysis were observed in any of the treatment groups.

[0368]

[0369] Example 7: Evaluation of synergistic effects of administration of the nanoparticles of the present invention (SNB-101) and radiation therapy in colon cancer and lung cancer treatment

[0370]

[0371] 7-1. Experimental materials

[0372] - BALB / c-nu mouse (Junabio), male, 6 weeks old

[0373] - Small cell lung cancer NCI-H69 cell line (ATCC, Cat.No.HTB-119)

[0374] - Non-small cell lung cancer A549 cell line (ATCC, Cat. No. CCL-185)

[0375] - Colorectal cancer HCT116 cell line (ATCC, Cat. No. CCL-247)

[0376] - Test drug: SNB-101 (SN BioScience Co., Ltd., SN-38 10 mg, Irinotecan HCl·3H 2 O 15.9 mg / vial)

[0377] - Vehicle: 50 ml of 5% glucose injection (CJ Healthcare, Insurance code 640001361)

[0378] - RPMI1640 medium (Gibco, 22400-089), F12K (Gibco, 21127-022), McCoy’s 5A (Gibco, 16600-082) supplemented with 10% fetal bovine serum (Gibco, Cat. No. 16000)

[0379] - 1% antibiotic-antimycotic (Gibco, Cat. No. 15240-062)

[0380] - Trypsin-EDTA (Gibco, Cat. No. 25200-072)

[0381] - Cold-DPBS (Gibco, Cat. No. 14200-075)

[0382] - Trypan blue (Gibco, Cat. No. 15250061)

[0383] - MycoAlert Mycoplasma detection kit (LONZA, Cat. No. LT-07-318)

[0384] - 150mm cell culture dish, Conical tubes, Pipettes

[0385] - Mouse ear tags, syringes, calipers

[0386]

[0387] 7-2.Test method

[0388] Frozen cell lines NCI-H69 (small cell lung cancer), HCT116 (colon cancer), and A549 (non-small cell lung cancer) held at the Anticancer T2B Foundation Center were thawed and cultured in RPMI1640, F12K, and McCoy's 5A medium - 10% FBS - 1% antibiotic-antimycotic at 37°C and 5% CO. 2 The thawed cells were cultured for more than one week, subcultured at 2-3 day intervals, and checked for viability, bacterial and yeast infection, and mycoplasma infection (MycoAlert Mycoplasma detection kit). Cells with a viability of 90% or more were used for transplantation. The cells were suspended in cold PBS after floating with trypsin-EDTA. 1×10 NCI-H69 cells were injected into the right thigh of a BALB / c-nu mouse. 7 cells / 100 μl and A549, HCT116 cells 1 × 10 6 The tumor formation and growth were periodically observed, and the tumor length was measured to calculate the tumor volume according to the following formula:

[0389] [Tumor volume = (a 2 b) / 2, a is the minor axis and b is the major axis]

[0390] The average tumor volume of all mice on the day of administration was 100±20mm 3 The mice were grouped by pair matching when the animals were in the above group. Each group was administered a drug according to the dosage regimen shown in Tables 17 and 18 below, and observed for 2 to 3 weeks.

[0391] The NCI-H69 and HCT116 models were administered once a week, and the A549 model was administered twice a week for 5 repeated doses. After drug administration, tumor volume and body weight were measured up to 21 days for the HCT116 model, up to 38 days for the NCI-H69 model, and up to 45 days for the A549 model.

[0392]

[0393] [Common to NCI-H69 (small cell lung cancer) and HCT116 (colon cancer) xenograft models]

[0394] [Table 17]

[0395] *:dose as SN-38 / Irinotecan HCl

[0396]

[0397] [A549 (non-small cell lung cancer) xenograft model]

[0398] [Table 18]

[0399] *:dose as SN-38 / Irinotecan HCl

[0400] **: In the A549 xenograft model, the SNB-101 administration dose was changed by mutual agreement during the study from 20 mg / kg (1st dose) to 30 mg / kg (2nd dose, occurrence of mortality) to 20 mg / kg (3rd to 5th doses).

[0401] ***: The administration frequency for the SNB-101 group was changed under mutual agreement during the course of the study to 4 times every 2 weeks (total of 5 times) BIW starting 9 days after the first administration.

[0402]

[0403] Tumor volume and body weight were measured twice a week (every 3 or 4 days) from the time of classification until the end of the study, and a tumor growth curve was created based on the tumor volume from the start of drug administration to the end of the study. When the tumor volume reached the limit set by the Institutional Animal Care and Use Committee (IACUC), the animals were euthanized in consultation with the client to terminate the study, and the tumors were excised and analyzed.

[0404]

[0405] 7-3.Test results

[0406] At the end of the experiment, tumor growth inhibition (TGI) relative to the vehicle was calculated using the above-mentioned formula 2.

[0407]

[0408] Table 19 shows the TGI by group for NCI-H69.

[0409] The tumor volume change curves are shown in FIG.

[0410] The tumor excision photograph is shown in FIG.

[0411] [Table 19]

[0412] For HCT116, the TGI by group is shown in Table 20.

[0413] The tumor volume change curves are shown in FIG.

[0414] The tumor excision photograph is shown in FIG.

[0415] [Table 20]

[0416] For A549, the TGI by group is shown in Table 21.

[0417] The tumor volume change curves are shown in FIG.

[0418] [Table 21]

[0419] As shown in Tables 19 to 21, the TGI relative to vehicle was significantly higher in the combined treatment group (G4) than in the SNB-101 alone treatment group (G2) and the radiation alone treatment group (G3). This confirmed that the combined treatment of SNB-101 and radiation had a highly synergistic effect when treating colon cancer and lung cancer.

[0420] Furthermore, no macroscopic side effects, weight loss, or toxicity in blood analysis were observed in any of the treatment groups.

[0421]

[0422] In summary, it was confirmed from the above-mentioned Examples 2 to 7 that a significant synergistic effect was achieved in cancer treatment when administered in combination or in combination with radiation therapy, as summarized in Table 22 below.

[0423] [Table 22]

Claims

1. Particles comprising SN-38 (7-ethyl-10-hydroxycamptothecin), irinotecan, and an amphiphilic block copolymer consisting of a hydrophobic block and a hydrophilic block; and Contains one or more antitumor agents selected from the group consisting of nab-paclitaxel as an active ingredient; The amphiphilic block copolymer is composed of an A-B or A-B-A block, (a) the A is a hydrophilic polymer and is monomethoxypolyethylene glycol; and (b) the B is a hydrophobic polymer and is poly-D,L-lactide or a derivative thereof. Pharmaceutical combination formulations for the treatment of pancreatic cancer.

2. Particles comprising SN-38 (7-ethyl-10-hydroxycamptothecin), irinotecan, and an amphiphilic block copolymer consisting of a hydrophobic block and a hydrophilic block; and Contains docetaxel as an active ingredient. The amphiphilic block copolymer is composed of an A-B or A-B-A block, (a) the A is a hydrophilic polymer and is monomethoxypolyethylene glycol; and (b) the B is a hydrophobic polymer and is poly-D,L-lactide or a derivative thereof. Pharmaceutical combination preparation for the treatment of gastric cancer.

3. Particles comprising SN-38 (7-ethyl-10-hydroxycamptothecin), irinotecan, and an amphiphilic block copolymer consisting of a hydrophobic block and a hydrophilic block; and It contains bevacizumab as an active ingredient. The amphiphilic block copolymer is composed of an A-B or A-B-A block, (a) the A is a hydrophilic polymer and is monomethoxypolyethylene glycol; and (b) the B is a hydrophobic polymer and is poly-D,L-lactide or a derivative thereof. Pharmaceutical combination preparation for the treatment of colon cancer.

4. The pharmaceutical combination formulation according to any one of claims 1 to 3, wherein the particles have an average diameter of 2 to 200 nm.

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