WO2024111897A1 - Pharmaceutical composition containing prd-2001 for preventing or treating cancer - Google Patents

Pharmaceutical composition containing prd-2001 for preventing or treating cancer Download PDF

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WO2024111897A1
WO2024111897A1 PCT/KR2023/016368 KR2023016368W WO2024111897A1 WO 2024111897 A1 WO2024111897 A1 WO 2024111897A1 KR 2023016368 W KR2023016368 W KR 2023016368W WO 2024111897 A1 WO2024111897 A1 WO 2024111897A1
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cancer
prd
cells
pancreatic
present
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French (fr)
Korean (ko)
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장현철
김수열
김종헌
우상명
이호
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국립암센터
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/337—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having four-membered rings, e.g. taxol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/4738—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/4745—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems condensed with ring systems having nitrogen as a ring hetero atom, e.g. phenantrolines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
    • A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
    • A61K31/704—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
    • A61K31/7052—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides
    • A61K31/706—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom
    • A61K31/7064—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines
    • A61K31/7068—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines having oxo groups directly attached to the pyrimidine ring, e.g. cytidine, cytidylic acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • the present invention relates to a composition for preventing or treating cancer containing PRD-2001 (ursonic acid), and more specifically, to a pharmaceutical composition for preventing or treating cancer containing PRD-2001 as an active ingredient, and an anticancer drug. It's about supplements.
  • PRD-2001 ursonic acid
  • cancer cells proliferate without limit, and a cell mass composed of such cancer cells is also called a tumor. These cancer cells infiltrate surrounding tissues and metastasize to other organs of the body, causing severe pain and ultimately causing death.
  • the number of cancer patients in Korea has continued to increase, increasing by about 44% over the past 10 years.
  • Internationally, the anticancer drug market has also increased, and has been reported to be worth about $100 billion annually.
  • pancreatic cancer is a fatal cancer with a 5-year survival rate of 1-4% and a median survival time of 5 months, showing the poorest prognosis among human cancers.
  • the prognosis is poor because curative resection with the expectation of cure is impossible at the time of diagnosis, and treatment mainly relies on chemotherapy.
  • the most commonly used anticancer drug combination for the anticancer treatment of pancreatic cancer is the FOLFIRNOX therapy, which combines four anticancer drugs (5-FU, leucovorin, irinotecan, and oxaliplatin), and the combination of gemcitabine and Abraxane (nab-paclitaxel). .
  • pancreatic cancer As a result of diligent efforts to select an anticancer agent effective for pancreatic cancer, it was confirmed that the growth of pancreatic cancer, breast cancer, liver cancer, and prostate cancer is inhibited by PRD-2001 (ursonic acid), and PRD-2001 and The anticancer synergy effect of co-administration of Irinotecan, Paclitaxel, Doxorubicin, or Gemcitabine was confirmed. Furthermore, it was confirmed that differentiation of undifferentiated cancer cells was promoted and cancer cell growth and DNA-damage repair were inhibited by PRD-2001, and the present invention was completed.
  • PRD-2001 ursonic acid
  • PRD-2001 ursonic acid
  • Gemcitabine Gemcitabine
  • the purpose of the present invention is to provide a pharmaceutical composition for preventing or treating cancer, or an anti-cancer adjuvant containing PRD-2001 (ursonic acid).
  • PRD-2001 ursonic acid
  • Another object of the present invention is to provide a pharmaceutical composition for preventing or treating cancer, or an anticancer adjuvant, containing PRD-2001 (ursonic acid) and an anticancer agent.
  • PRD-2001 ursonic acid
  • the present invention provides a pharmaceutical composition for preventing or treating pancreatic cancer, breast cancer, liver cancer, or prostate cancer, comprising PRD-2001 (ursonic acid) or a pharmaceutically acceptable salt thereof represented by the following formula (1) as an active ingredient. to provide.
  • PRD-2001 ursonic acid
  • a pharmaceutically acceptable salt thereof represented by the following formula (1) as an active ingredient. to provide.
  • the present invention provides an anti-cancer adjuvant for pancreatic cancer, breast cancer, liver cancer, or prostate cancer, comprising PRD-2001 (ursonic acid) or a pharmaceutically acceptable salt thereof represented by the following formula (1) as an active ingredient. do.
  • PRD-2001 ursonic acid
  • a pharmaceutically acceptable salt thereof represented by the following formula (1) as an active ingredient. do.
  • PRD-2001 can induce differentiation of undifferentiated cancer cells, inhibit cancer cell growth, and inhibit DNA damage repair in cancer cells.
  • the present invention relates to PRD-2001 (ursonic acid) represented by the following formula (1) or a pharmaceutically acceptable salt thereof; and irinotecan, gemcitabine, paclitaxel, and doxorubicin.
  • the present invention relates to PRD-2001 (ursonic acid) or a pharmaceutically acceptable salt thereof represented by the following formula (1) and irinotecan, gemcitabine, paclitaxel, and doxorubicin.
  • PRD-2001 ursonic acid
  • a pharmaceutically acceptable salt thereof represented by the following formula (1) and irinotecan, gemcitabine, paclitaxel, and doxorubicin.
  • an anti-cancer adjuvant containing as an active ingredient one or more anti-cancer agents selected from the group consisting of.
  • PRD-2001 can induce differentiation of undifferentiated cancer cells, inhibit cancer cell growth, and inhibit DNA damage repair in cancer cells.
  • PRD-2001 and the anticancer agent may be administered sequentially or simultaneously.
  • the cancer is any selected from the group consisting of pancreatic cancer, breast cancer, liver cancer, prostate cancer, colon cancer, lung cancer, stomach cancer, melanoma, prostate cancer, ovarian cancer, and glioblastoma. There may be more than one cancer.
  • PRD-2001 inhibits the growth of pancreatic cancer, breast cancer, prostate cancer, and liver cancer cells, and PRD-2001 and irinotecan, gemcitabine, paclitaxel, or doxorubicin
  • the anticancer synergy effect of combined administration was confirmed.
  • PRD-2001 promotes differentiation of undifferentiated cancer cells and inhibits cancer cell growth and DNA-damage repair, so the composition of the present invention can be usefully used as an effective anticancer agent.
  • Figure 1 shows data confirming the degree of cancer cell growth when various cancer cells were treated with PRD-2001, (A) pancreatic epithelial cell line, (B) renal cancer cell line, (C) pancreatic cancer cell line, (D) breast cancer cell line, (E) ) The degree of cancer cell growth was confirmed in the liver cancer cell line and (F) prostate cancer cell line.
  • Figure 2 shows data confirming (A) a decrease in the expression of the undifferentiated gene group, (B) a decrease in the expression of the cell cycle progression gene group, and (C) a decrease in the expression of the DNA damage repair-related gene group in cancer cells according to PRD-2001 treatment.
  • Figure 3 shows data confirming the degree of cancer cell growth when pancreatic cancer cell lines (A) PANC1 and (B) BXPC3 were treated with PRD-2001 and irinotecan alone or in combination.
  • Figure 4 shows data showing (A) change in tumor tissue volume, (B) observation of tumor tissue, and (C) change in tumor weight when irinotecan was treated alone at different concentrations in a cancer cell xenograft tumor model.
  • Figure 5 shows data observing (A) change in tumor tissue size, (B) change in tumor weight, and (C) change in mouse body weight when PRD-2001 and irinotecan were treated alone or in combination with a cancer cell xenograft tumor model.
  • Figure 6 shows PRD-2001 and (A) Gemcitabine, (B) Doxorubicin, (C) Paclitaxel, and (D) Fluorouracil (5-FU) in pancreatic cancer cell lines. This data confirms the anticancer synergy effect when administered in combination.
  • the present invention provides a pharmaceutical composition for preventing or treating pancreatic cancer, breast cancer, liver cancer or prostate cancer, comprising PRD-2001 (ursonic acid) or a pharmaceutically acceptable salt thereof represented by the following formula (1) as an active ingredient: It relates to pharmaceutical compositions.
  • the present invention provides an anti-cancer agent for pancreatic cancer, breast cancer, liver cancer or prostate cancer comprising PRD-2001 (ursonic acid) or a pharmaceutically acceptable salt thereof represented by the following formula (1) as an active ingredient. It's about supplements.
  • PRD-2001 ursonic acid
  • a pharmaceutically acceptable salt thereof represented by the following formula (1) as an active ingredient. It's about supplements.
  • PRD-2001 can induce differentiation of undifferentiated cancer cells, inhibit cancer cell growth, and inhibit DNA damage repair in cancer cells.
  • a normal pancreatic epithelial cell line HPNE
  • a kidney cancer cell line A498)
  • a pancreatic cancer cell line MIAPACA2
  • MDAMB231 a breast cancer cell line
  • HEP3B a liver cancer cell line
  • PC3 prostate cancer cell line
  • Cancer cells can be classified into mature cancers or undifferentiated cancers depending on their appearance. Differentiated cancers are close to mature normal cells, while undifferentiated cancers appear quite different from normal cells. Undifferentiated cancer divides faster than differentiated cancer, making it more likely to metastasize. When genetic abnormalities in cancer cells block differentiation, their growth and differentiation abilities become uncontrolled and they can divide indefinitely. By inducing normal differentiation in cancer cells and blocking their ability to proliferate, treatment that can partially change malignancy to benign is possible.
  • PRD-2001 can reduce the expression of undifferentiated genes in cancer cells, promote differentiation of cancer cells, and inhibit the growth of cancer cells by reducing cell cycle progression and inhibiting DNA damage repair.
  • PRD-2001 of the invention can be used as an effective anticancer agent or anticancer adjuvant.
  • the present invention relates to PRD-2001 (ursonic acid) represented by the following formula (1) or a pharmaceutically acceptable salt thereof; And it relates to a pharmaceutical composition for preventing or treating cancer, comprising as an active ingredient one or more anticancer agents selected from the group consisting of Irinotecan, gemcitabine, Paclitaxel, and Doxorubicin.
  • PRD-2001 ursonic acid
  • a pharmaceutical composition for preventing or treating cancer comprising as an active ingredient one or more anticancer agents selected from the group consisting of Irinotecan, gemcitabine, Paclitaxel, and Doxorubicin.
  • the present invention relates to PRD-2001 (ursonic acid) represented by the following formula (1) or a pharmaceutically acceptable salt thereof; and an anticancer adjuvant containing as an active ingredient one or more anticancer agents selected from the group consisting of Irinotecan, gemcitabine, Paclitaxel, and Doxorubicin.
  • PRD-2001 ursonic acid
  • an anticancer adjuvant containing as an active ingredient one or more anticancer agents selected from the group consisting of Irinotecan, gemcitabine, Paclitaxel, and Doxorubicin.
  • the cancer may be any one or more cancers selected from the group consisting of pancreatic cancer, breast cancer, liver cancer, prostate cancer, colon cancer, lung cancer, stomach cancer, melanoma, prostate cancer, ovarian cancer, and glioblastoma. there is.
  • composition of the present invention may be in various oral or parenteral dosage forms.
  • buffers e.g., saline or PBS
  • antioxidants e.g., bacteriostatic agents, chelating agents (e.g., EDTA or glutathione), fillers, extenders, binders, adjuvants (e.g., Aluminum hydroxide), suspending agents, thickening agents, wetting agents, disintegrants or surfactants, diluents or excipients.
  • bacteriostatic agents e.g., EDTA or glutathione
  • fillers e.g., extenders, binders
  • adjuvants e.g., Aluminum hydroxide
  • suspending agents thickening agents, wetting agents, disintegrants or surfactants, diluents or excipients.
  • Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations contain one or more compounds and at least one excipient, such as starch (corn starch, wheat starch, rice starch, potato). starch, etc.), calcium carbonate, sucrose, lactose, dextrose, sorbitol, mannitol, xylitol, erythritol, maltitol, cellulose, methyl cellulose, sodium carboxymethylcellulose, and hydroxypropylmethyl.
  • -It is prepared by mixing cellulose or gelatin.
  • tablets or dragees can be obtained by combining the active ingredient with solid excipients, grinding them, adding suitable auxiliaries, and processing them into a granule mixture.
  • Liquid preparations for oral administration include suspensions, oral solutions, emulsions, or syrups.
  • simple diluents such as water and liquid paraffin
  • they may contain various excipients such as wetting agents, sweeteners, fragrances, or preservatives. You can.
  • cross-linked polyvinylpyrrolidone, agar, alginic acid, or sodium alginate may be added as a disintegrant, and anti-coagulants, lubricants, wetting agents, fragrances, emulsifiers, and preservatives may be additionally included. .
  • Preparations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, or suppositories.
  • Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate.
  • injectable ester such as ethyl oleate.
  • As a base for suppositories witepsol, macrogol, tween 61, cacao, laurel, glycerol, gelatin, etc. can be used.
  • composition of the present invention can be administered orally or parenterally, and when administered parenterally, it can be applied externally to the skin; It can be formulated according to methods known in the art in the form of an injection for intraperitoneal, rectal, intravenous, intramuscular, subcutaneous, intrauterine intrathecal or intracerebrovascular injection.
  • suitable carriers include, but are not limited to, solvents or dispersion media including water, ethanol, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol, etc.), mixtures thereof, and/or vegetable oils. You can. More preferably, suitable carriers include Hanks' solution, Ringer's solution, phosphate buffered saline (PBS) containing triethanolamine, or isotonic solutions such as sterile water for injection, 10% ethanol, 40% propylene glycol, and 5% dextrose. etc. can be used.
  • solvents or dispersion media including water, ethanol, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol, etc.), mixtures thereof, and/or vegetable oils. You can. More preferably, suitable carriers include Hanks' solution, Ringer's solution, phosphate buffered saline (PBS) containing triethanolamine, or isotonic solutions such as
  • the injection may additionally contain various antibacterial and antifungal agents such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, etc.
  • various antibacterial and antifungal agents such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, etc.
  • the injection may additionally contain an isotonic agent such as sugar or sodium chloride.
  • composition of the present invention is administered in a pharmaceutically effective amount.
  • a pharmaceutically effective amount refers to an amount sufficient to treat a disease with a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level refers to the type of patient's disease, severity, activity of the drug, sensitivity to the drug, and administration time. , route of administration and excretion rate, duration of treatment, factors including concurrently used drugs, and other factors well known in the field of medicine.
  • the composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or multiple times.
  • the total effective amount of the composition of the present invention can be administered to the patient as a single dose, or may be administered by a fractionated treatment protocol in which multiple doses are administered over a long period of time. . Considering all of the above factors, it is important to administer an amount that can achieve maximum effect with the minimum amount without side effects, and this can be easily determined by a person skilled in the art.
  • the preferred dosage of the composition varies depending on the patient's condition, weight, degree of disease, drug form, administration route and period, but can be appropriately selected by a person skilled in the art, for example, 0.0001 to 2,000 mg/kg per day, Preferably, it can be administered at 0.001 to 2,000 mg/kg. Administration may be administered once a day, or may be administered in several divided doses. However, the scope of the present invention is not limited by the above dosage.
  • composition of the present invention can be used alone or in combination with surgery, radiation therapy, hormone therapy, chemotherapy, and methods using biological response modifiers.
  • the anticancer adjuvant of the present invention refers to all forms for increasing the anticancer effect of an anticancer agent or suppressing or improving the side effects of an anticancer agent.
  • the anti-cancer adjuvant of the present invention can be administered in combination with various types of anti-cancer drugs or anti-cancer adjuvants, and when administered in combination, it can exhibit the same level of anti-cancer treatment effect even if the anti-cancer agent is administered at a lower dosage than a typical anti-cancer agent, making anti-cancer safer. Treatment can be performed.
  • the anticancer adjuvant may be administered through any general route as long as it can reach the target tissue.
  • the anticancer adjuvant of the present invention may be administered intraperitoneally, intravenously, intramuscularly, subcutaneously, orally, pulmonaryly, or rectally, depending on the purpose, but is not limited thereto. Additionally, the anti-cancer adjuvant may be administered by any device capable of moving the active substance to target cells.
  • the anti-cancer adjuvant of the present invention can be preferably formulated as an anti-cancer adjuvant by containing one or more pharmaceutically acceptable carriers in addition to the active ingredient.
  • Carriers, excipients, or diluents that may be included in the anticancer treatment adjuvant of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, Including, but not limited to, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil.
  • the anticancer adjuvant of the present invention may be a preparation for oral or parenteral administration, and the description of the preparation is replaced with a description of the preparation of the pharmaceutical composition.
  • HPNE pancreatic epithelial cell line
  • A498 kidney cancer cell line
  • MIAPACA2 pancreatic cancer cell line
  • MDAMB231 breast cancer cell line
  • HEP3B liver cancer cell line
  • PC3 prostate cancer cell line
  • HPNE pancreatic endothelial cells
  • Human pancreatic adenocarcinoma cell line MIAPACA2, human breast cancer cell line MDAMB231, and human hepatocellular carcinoma cell line HEP3B were purchased from KCLB and cultured in the presence of 5% CO 2 in an incubator at 37°C using DMEM/high glucose culture medium containing 10% FBS.
  • Human prostate cancer cell line PC3 was purchased from KCLB and cultured in the presence of 5% CO 2 in an incubator at 37°C using RPMI culture medium containing 10% FBS.
  • Cell viability assay was performed to determine the effect of PRD-2001 on human pancreatic endothelial cells HPNE, human kidney cancer cell line (A498), human pancreatic cancer cell line MIAPACA2, human breast cancer cell line MDAMB231, human hepatocellular carcinoma cell line HEP3B, and human prostate cancer cell line PC3.
  • SRB assay was performed. Cells were cultured in a 96-well plate, treated with PRD-2001 at concentrations of 0, 1, and 5 ⁇ M, and cultured for 3 days. Cells were fixed at 4°C by adding 50 ⁇ l of 50% TCA solution, washed 5 times using DW, and the plate was dried.
  • the dried plate was treated with 100 ⁇ l of SRB solution at room temperature for 5 minutes, washed three times with 1% acetic acid, and then dried at room temperature. 100 ⁇ l of 10mM Tris solution was added to the dried plate and shaken for 30 minutes to dissolve SRB, and the absorbance was measured at 515 nm.
  • pancreatic cancer cell lines were treated with PRD-2001, the entire transcriptome analysis of pancreatic cancer cells was performed.
  • RNA sequencing was carried out. The quality and quantity of RNA were evaluated using Bioanalyzer 2100 and RNA 6000 Nano Labchips, and an RNA library was prepared using Truseq RNA-Seq Library Prep Kit-v2.
  • paired-end sequencing was performed using the NovaSeq6000 sequencing system.
  • Differentially expressed genes DEGs
  • TPM transcript per million mapped reads
  • GSEA Gene Set Enrichment Analysis
  • IPA Ingenuity pathway analysis
  • the human pancreatic adenocarcinoma cell line BXPC3 was purchased from KCLB and cultured in the presence of 5% CO 2 in an incubator at 37°C using RPMI culture medium containing 10% FBS.
  • pancreatic cancer cells showed an undifferentiated gene group [Figure 2A] and a gene group involved in cell cycle progression, which is the growth and division process of cancer cells [Figure 2B]. It was confirmed that this was downregulated. In addition, it was confirmed through IPA analysis [ Figure 2C] that the gene group downregulated in PRD-2001-treated pancreatic cancer cells was a gene that played a role in cancer cell growth and DNA damage repair mechanisms.
  • Example 3 Confirmation of anticancer synergy effect due to combined administration of PRD-2001 and irinotecan
  • PRD-2001 and irinotecan were administered alone or in combination to pancreatic cancer cell lines PANC1 and BXPC3, and then the degree of cancer cell growth was measured using a colony assay ( Clonogenic assay).
  • each pancreatic adenocarcinoma cell line was cultured in a 6-well plate, and treated alone or in combination with PRD-2001 at a concentration of 0 and 10 ⁇ M and irinotecan at a concentration of 0 and 0.5 ⁇ M, and cultured for 10 days.
  • the medium was replaced with a medium under the same conditions as above every ⁇ 3 days.
  • the cells were fixed and stained using 0.02% Crystal Violet solution, washed three times using PBS, and the plate was dried. The dried plate was imaged and then quantified using the Colony area plug-in of the 'Image J' program to confirm the results.
  • the results were analyzed using the 'Combenefit' program, and the synergy effect score was confirmed using the HSA model. A synergy effect score of 20 or higher was judged to have a synergistic effect.
  • Cancer cell growth inhibition rate (% compared to control group) following combined administration of PRD-2001 and irinotecan PANC1 BXPC3 Control 100.0000 100.0000 PRD-2001 10 ⁇ M 39.0981 61.0751 Irinotecan 0.5 ⁇ M 36.2929 73.5274 PRD-2001 10 ⁇ M+ Irinotecan 0.5 ⁇ M 6.1191 11.4837
  • a pancreatic cancer xenograft tumor model was created and the drug was administered.
  • mice 6-8 week old Balb/c nude mice were subcutaneously inoculated with MIAPACA2 cells ( 1 The animals were randomly divided into 3 groups with 5 animals per group as shown, and then PRD-2001 was administered. PRD-2001 was dissolved in 5% DMSO, 3% Cremophor EL, and 92% saline solution.
  • Tumor size was measured once a week using a caliper, and the weight of the mouse was also measured once a week. The size of the tumor was calculated using Equation 1 below.
  • Tumor size (short axis x short axis x long axis)/2
  • Tumor volume according to PRD-2001 administration Tumor volume ( mm3 ) 0 day 7 days 14 days 21 days Control 72.8016 148.4274 255.9562 388.6936 PRD-2001 50mg/kg 55.5426 122.5912 208.0494 344.134 PRD-2001 100 mg/kg 80.1236 108.784 96.0228 181.6902
  • Example 5 Confirmation of anticancer synergy effect due to combined administration of PRD-2001 and irinotecan in animal model
  • a pancreatic cancer xenograft tumor model was created and the drug was administered.
  • mice 6-8 week old Balb/c nude mice were subcutaneously inoculated with MIA PaCa-2 cells ( 1 Mice were randomly divided into 4 groups with 3 mice per group as follows, and then PRD-2001 and irinotecan were administered.
  • PRD-2001 was prepared by dissolving in 10% DMSO and 90% saline
  • irinotecan was prepared by dissolving in 4% DMSO and 96% saline.
  • Tumor size was measured twice a week using a caliper, and the body weight of the mouse was also measured twice a week.
  • the size of the tumor was calculated using Equation 1 below.
  • Tumor size (short axis x short axis x long axis)/2
  • Tumor volume following PRD-2001 and irinotecan co-administration Tumor volume ( mm3 ) 0 days 5 days 8 days 12 days 15 days Control 155.6883 258.2663 377.2575 648.5885 842.448 PRD-2001 50mg/kg 120.3473 214.1228 283.558 621.017 823.444 Irinotecan 20 mg/kg 108.0405 121.77 159.719 301.077 501.7455 PRD-2001 50 mg/kg + Irinotecan 20 mg/kg 78.99975 118.8765 134.514 176.176 149.2553
  • fluorouracil 5-FU, 1 ⁇ M
  • paclitaxel Paclitaxel, 1.25%
  • nM nM
  • Doxorubicin 5 nM
  • Gemcitabine 2.5 nM
  • PRD-2001 inhibits the growth of pancreatic cancer, breast cancer, prostate cancer, and liver cancer cells, and PRD-2001 and irinotecan, gemcitabine, paclitaxel, or doxorubicin Since the anti-cancer synergy effect of combined administration was confirmed, the composition of the present invention can be usefully used as an effective anti-cancer agent.

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  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

The present invention relates to a composition containing ursonic acid (PRD-2001) for preventing or treating cancer and, more specifically, to a pharmaceutical composition and an anti-cancer adjuvant for preventing or treating cancer, comprising PRD-2001 as an active ingredient. In the present invention, it has been identified that growth of pancreatic, breast, prostate and liver cancer cells is inhibited by PRD-2001, and anti-cancer synergistic effects obtained according to the co-administration of PRD-2001 with irinotecan, gemcitabine, paclitaxel or doxorubicin have been identified. Moreover, in the present invention, it has identified that differentiation of undifferentiated cancer cells is promoted and growth and DNA-damage repair of cancer cells are inhibited, and thus the composition of the present invention can be effectively used as an effective anti-cancer agent.

Description

PRD-2001을 포함하는 암 예방 또는 치료용 약학적 조성물Pharmaceutical composition for preventing or treating cancer containing PRD-2001
본 발명은 PRD-2001 (우르손산; ursonic acid)을 포함하는 암 예방 또는 치료용 조성물에 관한 것으로, 보다 상세하게는 PRD-2001을 유효성분으로 포함하는 암 예방 또는 치료용 약학적 조성물, 및 항암보조제에 관한 것이다.The present invention relates to a composition for preventing or treating cancer containing PRD-2001 (ursonic acid), and more specifically, to a pharmaceutical composition for preventing or treating cancer containing PRD-2001 as an active ingredient, and an anticancer drug. It's about supplements.
정상세포는 필요에 따라 규칙적이고 탄력적인 증식과 억제를 할 수 있는 반면에 암세포는 무제한의 증식을 하며, 이런 암세포로 구성된 세포덩어리를 종양이라고도 한다. 이러한 암세포는 주위의 조직으로 침투하고 신체의 다른 기관으로 전이가 되어 심각한 고통을 수반하고 결국 죽음을 초래한다. 의학의 발전에도 불구하고, 국내 암환자 발생자수는 지속적으로 증가하여 최근 10년간 약 44%가 증가하였으며, 국제적으로도 항암제 시장 역시 증가하여 연간 약 1000억 달러의 규모를 가지는 것으로 보고된 바 있다.While normal cells can regulate and flexibly proliferate and suppress as needed, cancer cells proliferate without limit, and a cell mass composed of such cancer cells is also called a tumor. These cancer cells infiltrate surrounding tissues and metastasize to other organs of the body, causing severe pain and ultimately causing death. Despite advances in medicine, the number of cancer patients in Korea has continued to increase, increasing by about 44% over the past 10 years. Internationally, the anticancer drug market has also increased, and has been reported to be worth about $100 billion annually.
특히, 췌장암은 5년 생존율이 1-4%, 중앙생존기간 5개월에 이르는 치명적인 암으로 인체의 암 중에서 가장 불량한 예후를 보이고 있다. 80-90% 환자에서 진단시 완치를 기대하는 근치적 절제가 불가능한 상태에서 발견되기 때문에 예후가 불량하고 치료는 주로 항암요법에 의존하고 있다. 췌장암의 항암치료에 가장 많이 사용되는 항암제 조합은 4가지 항암제 (5-FU, leucovorin, irinotecan, oxaliplatin)을 조합한 FOLFIRNOX 요법과 젬시타빈 (gemcitabine) 및 아브락산 (abraxane, nab-paclitaxel) 조합이 있다. In particular, pancreatic cancer is a fatal cancer with a 5-year survival rate of 1-4% and a median survival time of 5 months, showing the poorest prognosis among human cancers. In 80-90% of patients, the prognosis is poor because curative resection with the expectation of cure is impossible at the time of diagnosis, and treatment mainly relies on chemotherapy. The most commonly used anticancer drug combination for the anticancer treatment of pancreatic cancer is the FOLFIRNOX therapy, which combines four anticancer drugs (5-FU, leucovorin, irinotecan, and oxaliplatin), and the combination of gemcitabine and Abraxane (nab-paclitaxel). .
하지만, 상기 요법들을 이용한 항암치료에 대한 반응율은 15% 내외에 불과한 실정으로, 효과적으로 췌장암을 치료할 수 있는 약물의 개발이 요구되고 있다. However, the response rate to anti-cancer treatment using the above therapies is only about 15%, and there is a need for the development of a drug that can effectively treat pancreatic cancer.
이에, 본 발명에서는 췌장암에 효과적인 항암제를 선별하기 위해, 예의 노력한 결과 PRD-2001 (우르손산; ursonic acid)에 의해 췌장암, 유방암, 간암 및 전립선암의 성장이 억제되는 것을 확인하였으며, PRD-2001과 이리노테칸 (Irinotecan), 파클리탁셀 (Paclitaxel), 독소루비신 (Doxorubicin) 또는 젬시타빈 (Gemcitabine)의 병용투여에 따른 항암 시너지 효과를 확인하였다. 나아가, PRD-2001에 의해 미분화 암세포의 분화가 촉진되고, 암세포 성장 및 DNA-손상 복구가 억제되는 것을 확인하고, 본 발명을 완성하였다. Accordingly, in the present invention, as a result of diligent efforts to select an anticancer agent effective for pancreatic cancer, it was confirmed that the growth of pancreatic cancer, breast cancer, liver cancer, and prostate cancer is inhibited by PRD-2001 (ursonic acid), and PRD-2001 and The anticancer synergy effect of co-administration of Irinotecan, Paclitaxel, Doxorubicin, or Gemcitabine was confirmed. Furthermore, it was confirmed that differentiation of undifferentiated cancer cells was promoted and cancer cell growth and DNA-damage repair were inhibited by PRD-2001, and the present invention was completed.
따라서, 본 발명의 목적은 PRD-2001 (우르손산; ursonic acid)을 포함하는 암 예방 또는 치료용 약학적 조성물, 또는 항암 보조제를 제공하는 데 있다. Accordingly, the purpose of the present invention is to provide a pharmaceutical composition for preventing or treating cancer, or an anti-cancer adjuvant containing PRD-2001 (ursonic acid).
본 발명의 다른 목적은 PRD-2001 (우르손산; ursonic acid) 및 항암제를 포함하는 암 예방 또는 치료용 약학적 조성물, 또는 항암보조제를 제공하는 데 있다.Another object of the present invention is to provide a pharmaceutical composition for preventing or treating cancer, or an anticancer adjuvant, containing PRD-2001 (ursonic acid) and an anticancer agent.
상술한 목적을 달성하기 위해, To achieve the above-mentioned purpose,
본 발명은 하기 화학식 1로 표시되는 PRD-2001 (우르손산; ursonic acid) 또는 이의 약제학적으로 허용가능한 염을 유효성분으로 포함하는, 췌장암, 유방암, 간암 또는 전립선암 예방 또는 치료용 약학적 조성물을 제공한다. The present invention provides a pharmaceutical composition for preventing or treating pancreatic cancer, breast cancer, liver cancer, or prostate cancer, comprising PRD-2001 (ursonic acid) or a pharmaceutically acceptable salt thereof represented by the following formula (1) as an active ingredient. to provide.
또한, 본 발명은 하기 화학식 1로 표시되는 PRD-2001 (우르손산; ursonic acid) 또는 이의 약제학적으로 허용가능한 염을 유효성분으로 포함하는, 췌장암, 유방암, 간암 또는 전립선암에 대한 항암 보조제를 제공한다. In addition, the present invention provides an anti-cancer adjuvant for pancreatic cancer, breast cancer, liver cancer, or prostate cancer, comprising PRD-2001 (ursonic acid) or a pharmaceutically acceptable salt thereof represented by the following formula (1) as an active ingredient. do.
[화학식 1][Formula 1]
Figure PCTKR2023016368-appb-img-000001
Figure PCTKR2023016368-appb-img-000001
본 발명의 바람직한 일실시예에 있어서, 상기 PRD-2001은 미분화 암세포의 분화를 유도하고, 암세포 성장을 억제하며, 암세포의 DNA 손상 복구를 억제할 수 있다. In a preferred embodiment of the present invention, PRD-2001 can induce differentiation of undifferentiated cancer cells, inhibit cancer cell growth, and inhibit DNA damage repair in cancer cells.
다른 목적을 달성하기 위해, To achieve other purposes,
본 발명은 하기 화학식 1로 표시되는 PRD-2001 (우르손산; ursonic acid) 또는 이의 약제학적으로 허용가능한 염; 및 이리노테칸 (Irinotecan), 젬시타빈 (gemcitabine), 파클리탁셀 (Paclitaxel) 및 독소루비신 (Doxorubicin)으로 구성된 군에서 선택되는 어느 하나 이상의 항암제를 유효성분으로 포함하는 암 예방 또는 치료용 약학적 조성물을 제공한다. The present invention relates to PRD-2001 (ursonic acid) represented by the following formula (1) or a pharmaceutically acceptable salt thereof; and irinotecan, gemcitabine, paclitaxel, and doxorubicin.
또한, 본 발명은 하기 화학식 1로 표시되는 PRD-2001(우르손산; ursonic acid) 또는 이의 약제학적으로 허용가능한 염 및 이리노테칸 (Irinotecan), 젬시타빈 (gemcitabine), 파클리탁셀 (Paclitaxel) 및 독소루비신 (Doxorubicin)으로 구성된 군에서 선택되는 어느 하나 이상의 항암제를 유효성분으로 포함하는 항암 보조제를 제공한다.In addition, the present invention relates to PRD-2001 (ursonic acid) or a pharmaceutically acceptable salt thereof represented by the following formula (1) and irinotecan, gemcitabine, paclitaxel, and doxorubicin. Provided is an anti-cancer adjuvant containing as an active ingredient one or more anti-cancer agents selected from the group consisting of.
[화학식 1][Formula 1]
Figure PCTKR2023016368-appb-img-000002
Figure PCTKR2023016368-appb-img-000002
본 발명의 바람직한 일실시예에 있어서, 상기 PRD-2001은 미분화 암세포의 분화를 유도하고, 암세포 성장을 억제하며, 암세포의 DNA 손상 복구를 억제할 수 있다. In a preferred embodiment of the present invention, PRD-2001 can induce differentiation of undifferentiated cancer cells, inhibit cancer cell growth, and inhibit DNA damage repair in cancer cells.
본 발명의 바람직한 다른 일실시예에 있어서, 상기 PRD-2001 및 항암제는 순차적으로 또는 동시에 투여될 수 있다.In another preferred embodiment of the present invention, PRD-2001 and the anticancer agent may be administered sequentially or simultaneously.
본 발명의 바람직한 또 다른 일실시예에 있어서, 상기 암은 췌장암, 유방암, 간암, 전립선암, 대장암, 폐암, 위암, 흑색종, 전립선암, 난소암, 및 교모세포종 으로 이루어지는 군에서 선택되는 어느 하나 이상의 암일 수 있다.In another preferred embodiment of the present invention, the cancer is any selected from the group consisting of pancreatic cancer, breast cancer, liver cancer, prostate cancer, colon cancer, lung cancer, stomach cancer, melanoma, prostate cancer, ovarian cancer, and glioblastoma. There may be more than one cancer.
본 발명에서는 PRD-2001에 의해 췌장암, 유방암, 전립선암, 간암 세포의 성장을 억제하는 것을 확인하였으며, PRD-2001과 이리노테칸 (Irinotecan), 젬시타빈 (gemcitabine), 파클리탁셀 (Paclitaxel) 또는 독소루비신 (Doxorubicin)의 병용투여에 따른 항암 시너지 효과를 확인하였다. 나아가, 본 발명에서는 PRD-2001에 의해 미분화 암세포의 분화가 촉진되고, 암세포 성장 및 DNA-손상 복구가 억제되는 것을 확인하였으므로, 본 발명의 조성물은 효과적인 항암제로 유용하게 활용할 수 있다. In the present invention, it was confirmed that PRD-2001 inhibits the growth of pancreatic cancer, breast cancer, prostate cancer, and liver cancer cells, and PRD-2001 and irinotecan, gemcitabine, paclitaxel, or doxorubicin The anticancer synergy effect of combined administration was confirmed. Furthermore, in the present invention, it was confirmed that PRD-2001 promotes differentiation of undifferentiated cancer cells and inhibits cancer cell growth and DNA-damage repair, so the composition of the present invention can be usefully used as an effective anticancer agent.
도 1은 다양한 암세포에 PRD-2001을 처리하였을 때, 암세포 성장 정도를 확인한 데이터로, (A) 췌장상피 세포주, (B) 신장암 세포주, (C) 췌장암 세포주, (D) 유방암 세포주, (E) 간암 세포주, 및 (F) 전립선암 세포주에서 암세포 성장 정도를 확인하였다.Figure 1 shows data confirming the degree of cancer cell growth when various cancer cells were treated with PRD-2001, (A) pancreatic epithelial cell line, (B) renal cancer cell line, (C) pancreatic cancer cell line, (D) breast cancer cell line, (E) ) The degree of cancer cell growth was confirmed in the liver cancer cell line and (F) prostate cancer cell line.
도 2는 PRD-2001 처리에 따른 암세포 내 (A) 미분화 유전자군 발현 감소, (B) 세포주기진행 유전자군 발현 감소, 및 (C) DNA 손상 복구 관련 유전자군 발현 감소를 확인한 데이터이다.Figure 2 shows data confirming (A) a decrease in the expression of the undifferentiated gene group, (B) a decrease in the expression of the cell cycle progression gene group, and (C) a decrease in the expression of the DNA damage repair-related gene group in cancer cells according to PRD-2001 treatment.
도 3은 췌장암 세포주인 (A) PANC1 및 (B) BXPC3에 PRD-2001 및 이리노테칸을 단독 또는 병용 처리하였을 때, 암세포 성장 정도를 확인한 데이터이다.Figure 3 shows data confirming the degree of cancer cell growth when pancreatic cancer cell lines (A) PANC1 and (B) BXPC3 were treated with PRD-2001 and irinotecan alone or in combination.
도 4는 암세포 이종이식 종양 모델에 이리노테칸을 농도를 다르게 하여 단독처리 하였을 때, (A) 종양 조직 부피 변화, (B) 종양 조직 관찰 및 (C) 종양의 무게 변화를 관찰한 데이터이다.Figure 4 shows data showing (A) change in tumor tissue volume, (B) observation of tumor tissue, and (C) change in tumor weight when irinotecan was treated alone at different concentrations in a cancer cell xenograft tumor model.
도 5는 암세포 이종이식 종양 모델에 PRD-2001 및 이리노테칸을 단독 또는 병용처리 하였을 때, (A) 종양 조직 크기 변화, (B) 종양의 무게 변화, (C) 마우스 체중 변화를 관찰한 데이터이다.Figure 5 shows data observing (A) change in tumor tissue size, (B) change in tumor weight, and (C) change in mouse body weight when PRD-2001 and irinotecan were treated alone or in combination with a cancer cell xenograft tumor model.
도 6은 PRD-2001과 (A) 젬시타빈 (Gemcitabine), (B) 독소루비신 (Doxorubicin), (C) 파클리탁셀 (Paclitaxel), 및 (D) 플루오로우라실 (fluorouracil, 5-FU)을 췌장암 세포주에 병용투여하였을 때, 항암 시너지 효과를 확인한 데이터이다. Figure 6 shows PRD-2001 and (A) Gemcitabine, (B) Doxorubicin, (C) Paclitaxel, and (D) Fluorouracil (5-FU) in pancreatic cancer cell lines. This data confirms the anticancer synergy effect when administered in combination.
이하, 발명을 상세하게 설명한다. Hereinafter, the invention will be described in detail.
본 발명은 일관점에서, 하기 화학식 1로 표시되는 PRD-2001 (우르손산; ursonic acid) 또는 이의 약제학적으로 허용가능한 염을 유효성분으로 포함하는, 췌장암, 유방암, 간암 또는 전립선암 예방 또는 치료용 약학적 조성물에 관한 것이다.Consistently, the present invention provides a pharmaceutical composition for preventing or treating pancreatic cancer, breast cancer, liver cancer or prostate cancer, comprising PRD-2001 (ursonic acid) or a pharmaceutically acceptable salt thereof represented by the following formula (1) as an active ingredient: It relates to pharmaceutical compositions.
본 발명은 다른 일관점에서, 하기 화학식 1로 표시되는 PRD-2001 (우르손산; ursonic acid) 또는 이의 약제학적으로 허용가능한 염을 유효성분으로 포함하는, 췌장암, 유방암, 간암 또는 전립선암에 대한 항암 보조제에 관한 것이다.In another consistent sense, the present invention provides an anti-cancer agent for pancreatic cancer, breast cancer, liver cancer or prostate cancer comprising PRD-2001 (ursonic acid) or a pharmaceutically acceptable salt thereof represented by the following formula (1) as an active ingredient. It's about supplements.
[화학식 1][Formula 1]
Figure PCTKR2023016368-appb-img-000003
Figure PCTKR2023016368-appb-img-000003
본 발명에 있어서, 상기 PRD-2001은 미분화 암세포의 분화를 유도하고, 암세포 성장을 억제하며, 암세포의 DNA 손상 복구를 억제시킬 수 있다. In the present invention, PRD-2001 can induce differentiation of undifferentiated cancer cells, inhibit cancer cell growth, and inhibit DNA damage repair in cancer cells.
본 발명의 구체적인 일실시예에 있어서, 정상 췌장상피 세포주 (HPNE), 신장암 세포주 (A498), 췌장암 세포주 (MIAPACA2), 유방암 세포주 (MDAMB231), 간암 세포주 (HEP3B) 및 전립선암 세포주 (PC3)에 PRD-2001을 처리한 결과, 췌장암 세포주, 유방암 세포주, 간암 세포주 및 전립선암 세포주만 성장이 억제되었으며, 정상 췌장상피 세포주 및 신장암 세포주에 대해서는 성장 억제 효과가 없는 것을 확인하였다 [도 1].In a specific embodiment of the present invention, a normal pancreatic epithelial cell line (HPNE), a kidney cancer cell line (A498), a pancreatic cancer cell line (MIAPACA2), a breast cancer cell line (MDAMB231), a liver cancer cell line (HEP3B), and a prostate cancer cell line (PC3). As a result of treatment with PRD-2001, the growth of only pancreatic cancer cell lines, breast cancer cell lines, liver cancer cell lines, and prostate cancer cell lines was inhibited, and it was confirmed that there was no growth inhibition effect on normal pancreatic epithelial cell lines and renal cancer cell lines [Figure 1].
본 발명의 구체적인 다른 일실시예에서, PRD-2001에 의한 암세포 사멸 작용 기전을 확인한 결과, PRD-2001에 의해, 암세포 내 미분화 유전자군 및 세포주기 진행 유전자군의 발현이 감소한 것을 확인하였다. 또한, 암세포의 DNA 손상 복구 관련 유전자군 발현 역시 감소한 것을 확인하였다 [도 2]. In another specific embodiment of the present invention, as a result of confirming the mechanism of cancer cell death by PRD-2001, it was confirmed that the expression of undifferentiated genes and cell cycle progression genes in cancer cells was reduced by PRD-2001. In addition, it was confirmed that the expression of genes related to DNA damage repair in cancer cells was also decreased [Figure 2].
암세포는 생긴 양상에 따라 분화암 또는 미분화암으로 성숙도를 나눌 수 있으며, 분화암은 성숙한 정상세포에 가까운 양상이고, 미분화암은 정상세포와는 상당히 다른 모습을 보인다. 미분화암일수록 분화암에 비해 분열속도가 빨라 전이가 될 가능성이 높다. 암세포의 유전적 비정상이 분화를 차단하면, 성장과 분화 능력이 조절되지 않아 무한정 분열할 수 있게 된다. 암세포에서 정상적인 분화를 유도하여 암세포의 증식 능력을 차단하면 부분적으로 악성을 양성으로 바꿀 수 있는 치료가 가능하다. Cancer cells can be classified into mature cancers or undifferentiated cancers depending on their appearance. Differentiated cancers are close to mature normal cells, while undifferentiated cancers appear quite different from normal cells. Undifferentiated cancer divides faster than differentiated cancer, making it more likely to metastasize. When genetic abnormalities in cancer cells block differentiation, their growth and differentiation abilities become uncontrolled and they can divide indefinitely. By inducing normal differentiation in cancer cells and blocking their ability to proliferate, treatment that can partially change malignancy to benign is possible.
즉, 본 발명에서는 PRD-2001에 의해 암세포의 미분화 유전자군 발현이 감소되어 암세포의 분화가 촉진되고, 세포주기 진행 감소 및 DNA 손상 복구 억제를 통해 암세포의 성장을 억제시킬 수 있는 것을 확인하였으므로, 본 발명의 PRD-2001은 효과적인 항암제 또는 항암보조제로 활용될 수 있다. In other words, the present invention confirmed that PRD-2001 can reduce the expression of undifferentiated genes in cancer cells, promote differentiation of cancer cells, and inhibit the growth of cancer cells by reducing cell cycle progression and inhibiting DNA damage repair. PRD-2001 of the invention can be used as an effective anticancer agent or anticancer adjuvant.
본 발명은 또 다른 관점에서, 하기 화학식 1로 표시되는 PRD-2001 (우르손산; ursonic acid) 또는 이의 약제학적으로 허용가능한 염; 및 이리노테칸 (Irinotecan), 젬시타빈 (gemcitabine), 파클리탁셀 (Paclitaxel) 및 독소루비신 (Doxorubicin)으로 구성된 군에서 선택되는 어느 하나 이상의 항암제를 유효성분으로 포함하는 암 예방 또는 치료용 약학적 조성물에 관한 것이다. From another aspect, the present invention relates to PRD-2001 (ursonic acid) represented by the following formula (1) or a pharmaceutically acceptable salt thereof; And it relates to a pharmaceutical composition for preventing or treating cancer, comprising as an active ingredient one or more anticancer agents selected from the group consisting of Irinotecan, gemcitabine, Paclitaxel, and Doxorubicin.
본 발명은 또 다른 관점에서, 하기 화학식 1로 표시되는 PRD-2001 (우르손산; ursonic acid) 또는 이의 약제학적으로 허용가능한 염; 및 이리노테칸 (Irinotecan), 젬시타빈 (gemcitabine), 파클리탁셀 (Paclitaxel) 및 독소루비신 (Doxorubicin)으로 구성된 군에서 선택되는 어느 하나 이상의 항암제를 유효성분으로 포함하는 항암 보조제에 관한 것이다.From another aspect, the present invention relates to PRD-2001 (ursonic acid) represented by the following formula (1) or a pharmaceutically acceptable salt thereof; and an anticancer adjuvant containing as an active ingredient one or more anticancer agents selected from the group consisting of Irinotecan, gemcitabine, Paclitaxel, and Doxorubicin.
[화학식 1][Formula 1]
Figure PCTKR2023016368-appb-img-000004
Figure PCTKR2023016368-appb-img-000004
본 발명의 구체적인 일실시예에서, 췌장암 세포주인 PANC1 및 BxPC3에 PRD-2001 및 이리노테칸을 병용투여한 결과, 암세포 성장 억제 효능이 크게 증가한 것을 확인하였으며, PANC1 및 BxPC3 각각에 대한 병용투여에 의한 시너지 점수 (Synergy score)는 30 및 50인 것으로 나타났다 [도 3]. In a specific embodiment of the present invention, as a result of combined administration of PRD-2001 and irinotecan to pancreatic cancer cell lines PANC1 and BxPC3, it was confirmed that the cancer cell growth inhibitory effect was significantly increased, and the synergy score by combined administration for each of PANC1 and BxPC3 (Synergy score) was found to be 30 and 50 [Figure 3].
또한, 암세포 이종이식 종양 모델에 PRD-2001을 단독투여한 결과, PRD-2001 농도에 따라 대조군에 비해 종양조직의 부피 및 무게가 현저히 감소되는 것을 확인하였다 [도 4].In addition, when PRD-2001 was administered alone to a cancer cell xenograft tumor model, it was confirmed that the volume and weight of tumor tissue were significantly reduced compared to the control group depending on the concentration of PRD-2001 [Figure 4].
암세포 이종이식 종양 모델에서 PRD-2001 및 이리노테칸을 병용투여한 결과, 대조군 및 단독투여군에 비해 종양조직 부피 및 부게가 현저하게 감소하였고, 이 때 마우스의 체중에는 변화는 없는 것을 확인하였다 [도 5].As a result of combined administration of PRD-2001 and irinotecan in a cancer cell xenograft tumor model, tumor tissue volume and mass were significantly reduced compared to the control and single administration groups, and there was no change in the body weight of the mouse [Figure 5] .
본 발명의 구체적인 다른 일실시예에서, 이리노테칸 이외에 플루오로우라실 (fluorouracil, 5-FU), 파클리탁셀 (Paclitaxel), 독소루비신 (Doxorubicin), 또는 젬시타빈 (Gemcitabine)을 PRD-2001과 췌장암 세포주에 병용투여한 결과, 젬시타빈 (Gemcitabine), 독소루비신 (Doxorubicin), 및 파클리탁셀 (Paclitaxel)이 PRD-2001과 병용투여에 따른 항암 시너지 효과를 보이는 것을 확인하였다 [도 6]. In another specific embodiment of the present invention, in addition to irinotecan, fluorouracil (5-FU), paclitaxel, doxorubicin, or gemcitabine was administered in combination with PRD-2001 and a pancreatic cancer cell line. As a result, it was confirmed that Gemcitabine, Doxorubicin, and Paclitaxel showed an anticancer synergy effect when administered in combination with PRD-2001 [Figure 6].
본 발명에 있어서, 상기 암은 상기 암은 췌장암, 유방암, 간암, 전립선암, 대장암, 폐암, 위암, 흑색종, 전립선암, 난소암, 및 교모세포종으로 이루어지는 군에서 선택되는 어느 하나 이상의 암일 수 있다.In the present invention, the cancer may be any one or more cancers selected from the group consisting of pancreatic cancer, breast cancer, liver cancer, prostate cancer, colon cancer, lung cancer, stomach cancer, melanoma, prostate cancer, ovarian cancer, and glioblastoma. there is.
본 발명의 조성물은 경구 또는 비경구의 여러 가지 제형일 수 있다. 상기 조성물을 제형화할 경우에는 하나 이상의 완충제 (예를 들어, 식염수 또는 PBS), 항산화제, 정균제, 킬레이트화제 (예를 들어, EDTA 또는 글루타치온), 충진제, 증량제, 결합제, 아쥬반트 (예를 들어, 알루미늄 하이드록사이드), 현탁제, 농후제 습윤제, 붕해제 또는 계면활성제, 희석제 또는 부형제를 사용하여 조제될 수 있다. The composition of the present invention may be in various oral or parenteral dosage forms. When formulating the composition, one or more buffers (e.g., saline or PBS), antioxidants, bacteriostatic agents, chelating agents (e.g., EDTA or glutathione), fillers, extenders, binders, adjuvants (e.g., Aluminum hydroxide), suspending agents, thickening agents, wetting agents, disintegrants or surfactants, diluents or excipients.
경구투여를 위한 고형제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형제제는 하나 이상의 화합물에 적어도 하나 이상의 부형제 예를 들면, 전분(옥수수 전분, 밀 전분, 쌀 전분, 감자 전분 등 포함), 칼슘카보네이트 (calcium carbonate), 수크로스 (sucrose), 락토오스 (lactose), 덱스트로오스, 솔비톨, 만니톨, 자일리톨, 에리스리톨 말티톨, 셀룰로즈, 메틸 셀룰로즈, 나트륨 카르복시메틸셀룰로오즈 및 하이드록시프로필메틸-셀룰로즈 또는 젤라틴 등을 섞어 조제된다. 예컨대, 활성성분을 고체 부형제와 배합한 다음 이를 분쇄하고 적합한 보조제를 첨가한 후 과립 혼합물로 가공함으로써 정제 또는 당의정제를 수득할 수 있다. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations contain one or more compounds and at least one excipient, such as starch (corn starch, wheat starch, rice starch, potato). starch, etc.), calcium carbonate, sucrose, lactose, dextrose, sorbitol, mannitol, xylitol, erythritol, maltitol, cellulose, methyl cellulose, sodium carboxymethylcellulose, and hydroxypropylmethyl. -It is prepared by mixing cellulose or gelatin. For example, tablets or dragees can be obtained by combining the active ingredient with solid excipients, grinding them, adding suitable auxiliaries, and processing them into a granule mixture.
또한, 단순한 부형제 이외에 스테아린산 마그네슘, 탈크 등과 같은 윤활제들도 사용된다. 경구투여를 위한 액상 제제로는 현탁제, 내용액제, 유제 또는 시럽제 등이 해당되는데, 흔히 사용되는 단순 희석제인 물, 리퀴드 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제 또는 보존제 등이 포함될 수 있다. 또한, 경우에 따라 가교결합 폴리비닐피롤리돈, 한천, 알긴산 또는 나트륨 알기네이트 등을 붕해제로 첨가할 수 있으며, 항응집제, 윤활제, 습윤제, 향료, 유화제 및 방부제 등을 추가로 포함할 수 있다.Additionally, in addition to simple excipients, lubricants such as magnesium stearate, talc, etc. are also used. Liquid preparations for oral administration include suspensions, oral solutions, emulsions, or syrups. In addition to the commonly used simple diluents such as water and liquid paraffin, they may contain various excipients such as wetting agents, sweeteners, fragrances, or preservatives. You can. In addition, in some cases, cross-linked polyvinylpyrrolidone, agar, alginic acid, or sodium alginate may be added as a disintegrant, and anti-coagulants, lubricants, wetting agents, fragrances, emulsifiers, and preservatives may be additionally included. .
비경구 투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁용제, 유제, 동결건조제제 또는 좌제 등이 포함된다. 비수성용제 및 현탁용제로는 프로필렌글리콜 (propylene glycol), 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔 (witepsol), 마크로골, 트윈 (tween) 61, 카카오지, 라우린지, 글리세롤, 젤라틴 등이 사용될 수 있다.Preparations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, or suppositories. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate. As a base for suppositories, witepsol, macrogol, tween 61, cacao, laurel, glycerol, gelatin, etc. can be used.
본 발명의 조성물은 경구 또는 비경구로 투여될 수 있으며, 비경구 투여시 피부외용; 복강내, 직장, 정맥, 근육, 피하, 자궁내 경막 또는 뇌혈관내 주사하는 주사제의 형태로 당업계에 공지된 방법에 따라 제형화할 수 있다.The composition of the present invention can be administered orally or parenterally, and when administered parenterally, it can be applied externally to the skin; It can be formulated according to methods known in the art in the form of an injection for intraperitoneal, rectal, intravenous, intramuscular, subcutaneous, intrauterine intrathecal or intracerebrovascular injection.
상기 주사제의 경우에는 반드시 멸균되어야 하며 박테리아 및 진균과 같은 미생물의 오염으로부터 보호되어야 한다. 주사제의 경우 적합한 담체의 예로는 이에 한정되지는 않으나, 물, 에탄올, 폴리올 (예를 들어, 글리세롤, 프로필렌 글리콜 및 액체 폴리에틸렌 글리콜 등), 이들의 혼합물 및/또는 식물유를 포함하는 용매 또는 분산매질일 수 있다. 보다 바람직하게는, 적합한 담체로는 행크스 용액, 링거 용액, 트리에탄올 아민이 함유된 PBS (phosphate buffered saline) 또는 주사용 멸균수, 10% 에탄올, 40% 프로필렌 글리콜 및 5% 덱스트로즈와 같은 등장 용액 등을 사용할 수 있다. 상기 주사제를 미생물 오염으로부터 보호하기 위해서는 파라벤, 클로로부탄올, 페놀, 소르빈산, 티메로살 등과 같은 다양한 항균제 및 항진균제를 추가로 포함할 수 있다. 또한, 상기 주사제는 대부분의 경우 당 또는 나트륨 클로라이드와 같은 등장화제를 추가로 포함할 수 있다.The above injections must be sterilized and protected from contamination by microorganisms such as bacteria and fungi. For injections, examples of suitable carriers include, but are not limited to, solvents or dispersion media including water, ethanol, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol, etc.), mixtures thereof, and/or vegetable oils. You can. More preferably, suitable carriers include Hanks' solution, Ringer's solution, phosphate buffered saline (PBS) containing triethanolamine, or isotonic solutions such as sterile water for injection, 10% ethanol, 40% propylene glycol, and 5% dextrose. etc. can be used. In order to protect the injection from microbial contamination, it may additionally contain various antibacterial and antifungal agents such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, etc. In addition, in most cases, the injection may additionally contain an isotonic agent such as sugar or sodium chloride.
본 발명의 조성물은 약제학적으로 유효한 양으로 투여한다. 약제학적으로 유효한 양은 의학적 치료에 적용 가능한 합리적인 수혜/위험 비율로 질환을 치료하기에 충분한 양을 의미하며, 유효용량 수준은 환자의 질환의 종류, 중증도, 약물의 활성, 약물에 대한 민감도, 투여 시간, 투여 경로 및 배출 비율, 치료기간, 동시 사용되는 약물을 포함한 요소 및 기타 의학 분야에 잘 알려진 요소에 따라 결정될 수 있다. 본 발명의 조성물은 개별 치료제로 투여하거나 다른 치료제와 병용하여 투여될 수 있고 종래의 치료제와는 순차적 또는 동시에 투여될 수 있으며, 단일 또는 다중 투여될 수 있다. 즉, 본 발명의 조성물의 총 유효량은 단일 투여량 (single dose)으로 환자에게 투여될 수 있으며, 다중 투여량 (multiple dose)으로 장기간 투여되는 분할 치료 방법 (fractionated treatment protocol)에 의해 투여될 수 있다. 상기한 요소들을 모두 고려하여 부작용 없이 최소한의 양으로 최대 효과를 얻을 수 있는 양을 투여하는 것이 중요하며, 이는 당업자에 의해 용이하게 결정될 수 있다.The composition of the present invention is administered in a pharmaceutically effective amount. A pharmaceutically effective amount refers to an amount sufficient to treat a disease with a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level refers to the type of patient's disease, severity, activity of the drug, sensitivity to the drug, and administration time. , route of administration and excretion rate, duration of treatment, factors including concurrently used drugs, and other factors well known in the field of medicine. The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or multiple times. That is, the total effective amount of the composition of the present invention can be administered to the patient as a single dose, or may be administered by a fractionated treatment protocol in which multiple doses are administered over a long period of time. . Considering all of the above factors, it is important to administer an amount that can achieve maximum effect with the minimum amount without side effects, and this can be easily determined by a person skilled in the art.
상기 조성물의 바람직한 투여량은 환자의 상태, 체중, 질병의 정도, 약물형태, 투여경로 및 기간에 따라 다르지만, 당업자에 의해 적절하게 선택될 수 있으며, 예컨대 1일 0.0001 내지 2,000 mg/kg으로, 더욱 바람직하게는 0.001 내지 2,000 mg/kg으로 투여할 수 있다. 투여는 하루에 한 번 투여할 수도 있고, 수회 나누어서 투여할 수도 있다. 다만, 상기 투여량에 의해서 본 발명의 범위를 한정하는 것은 아니다.The preferred dosage of the composition varies depending on the patient's condition, weight, degree of disease, drug form, administration route and period, but can be appropriately selected by a person skilled in the art, for example, 0.0001 to 2,000 mg/kg per day, Preferably, it can be administered at 0.001 to 2,000 mg/kg. Administration may be administered once a day, or may be administered in several divided doses. However, the scope of the present invention is not limited by the above dosage.
본 발명의 조성물은 단독으로, 또는 수술, 방사선 치료, 호르몬 치료, 화학 치료 및 생물학적 반응 조절제를 사용하는 방법들과 병용하여 사용할 수 있다.The composition of the present invention can be used alone or in combination with surgery, radiation therapy, hormone therapy, chemotherapy, and methods using biological response modifiers.
본 발명의 항암보조제는 항암제의 항암효과를 증대시키거나 항암제의 부작용을 억제 또는 개선시키기 위한 모든 형태를 의미한다. 본 발명의 항암보조제는 다양한 종류의 항암제 또는 항암보조제와 병용투여될 수 있으며, 병용투여시 통상적인 항암제의 투여량보다 낮은 수준으로 항암제를 투여하더라도 동등한 수준의 항암치료효과를 나타낼 수 있으므로 보다 안전한 항암치료를 수행할 수 있다.The anticancer adjuvant of the present invention refers to all forms for increasing the anticancer effect of an anticancer agent or suppressing or improving the side effects of an anticancer agent. The anti-cancer adjuvant of the present invention can be administered in combination with various types of anti-cancer drugs or anti-cancer adjuvants, and when administered in combination, it can exhibit the same level of anti-cancer treatment effect even if the anti-cancer agent is administered at a lower dosage than a typical anti-cancer agent, making anti-cancer safer. Treatment can be performed.
상기 항암보조제의 투여 경로는 목적 조직에 도달할 수 있는 한 어떠한 일반적인 경로를 통하여 투여될 수 있다. 본 발명의 항암보조제는 목적하는 바에 따라 복강 내 투여, 정맥 내 투여, 근육 내 투여, 피하 투여, 경구 투여, 폐 내 투여, 직장 내 투여될 수 있으나, 이에 제한되지는 않는다. 또한, 상기 항암보조제는 활성 물질이 표적 세포로 이동할 수 있는 임의의 장치에 의해 투여될 수 있다.The anticancer adjuvant may be administered through any general route as long as it can reach the target tissue. The anticancer adjuvant of the present invention may be administered intraperitoneally, intravenously, intramuscularly, subcutaneously, orally, pulmonaryly, or rectally, depending on the purpose, but is not limited thereto. Additionally, the anti-cancer adjuvant may be administered by any device capable of moving the active substance to target cells.
본 발명의 항암보조제는 투여를 위해서 유효 성분 이외에 추가로 약제학적으로 허용 가능한 담체를 1종 이상 포함하여 항암보조제로 바람직하게 제제화할 수 있다. 본 발명의 항암치료 보조제에 포함될 수 있는 담체, 부형제 또는 희석제로는, 락토즈, 덱스트로즈, 수크로스, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로즈, 메틸 셀룰로즈, 미정질 셀룰로스, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유를 포함하나 이에 제한되는 것은 아니다.For administration, the anti-cancer adjuvant of the present invention can be preferably formulated as an anti-cancer adjuvant by containing one or more pharmaceutically acceptable carriers in addition to the active ingredient. Carriers, excipients, or diluents that may be included in the anticancer treatment adjuvant of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, Including, but not limited to, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil.
본 발명의 항암보조제는 경구 또는 비경구 투여를 위한 제제일 수 있으며, 제제에 대한 설명은 상기 약학적 조성물의 제제에 대한 기재로 대신한다. The anticancer adjuvant of the present invention may be a preparation for oral or parenteral administration, and the description of the preparation is replaced with a description of the preparation of the pharmaceutical composition.
이하, 실시예를 통하여 본 발명을 보다 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 예시하기 위한 것으로서, 본 발명의 범위가 이들 실시예에 의하여 제한되는 것으로 해석하지 않는 것은 해당 기술분야에서 통상의 지식을 가진 자에 있어서 자명한 것이다.Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, and it is obvious to those skilled in the art that the scope of the present invention should not be construed as limited by these examples.
실시예 1: PRD-2001 항암 효과 확인Example 1: Confirmation of PRD-2001 anticancer effect
본 발명에서는 PRD-2001의 항암 효과를 확인하기 위해, 정상 췌장상피 세포주 (HPNE), 신장암 세포주 (A498), 췌장암 세포주 (MIAPACA2), 유방암 세포주 (MDAMB231), 간암 세포주 (HEP3B) 및 전립선암 세포주 (PC3)에 PRD-2001을 농도별로 (0, 1, 5μM) 처리한 다음, 각각의 암세포 성장 정도를 확인하였다.In the present invention, to confirm the anti-cancer effect of PRD-2001, normal pancreatic epithelial cell line (HPNE), kidney cancer cell line (A498), pancreatic cancer cell line (MIAPACA2), breast cancer cell line (MDAMB231), liver cancer cell line (HEP3B), and prostate cancer cell line. (PC3) was treated with PRD-2001 at various concentrations (0, 1, 5 μM), and then the degree of growth of each cancer cell was checked.
구체적으로, 인간 췌장 내피세포인 HPNE는 ATCC에서 구매하였으며, HPNE 세포주는 글루코스 (glucose)가 없는 DMEM 배양 배지에 5% FBS (Fetal bovine serum), 10 ng/ml 인간 재조합 EGF, 5.5 mM D-글루코스, 750 ng/ml 퓨로마이신 (puromycin)을 추가한 배지를 이용하여 37℃ 인큐베이터에서 5% CO2 존재 하에 배양하였다. 인간 췌관선암종 세포주 MIAPACA2, 인간 유방암 세포주 MDAMB231, 인간 간세포암 세포주 HEP3B는 KCLB에서 구매하였으며, 10% FBS를 포함하는 DMEM/high glucose 배양 배지를 이용하여 37℃ 인큐베이터에서 5% CO2 존재 하에 배양하였다. 인간 전립선암 세포주 PC3는 KCLB에서 구매하였으며, 10% FBS를 포함하는 RPMI 배양 배지를 이용하여 37℃ 인큐베이터에서 5% CO2 존재 하에 배양하였다.Specifically, human pancreatic endothelial cells, HPNE, were purchased from ATCC, and the HPNE cell line was cultured in glucose-free DMEM culture medium supplemented with 5% FBS (Fetal bovine serum), 10 ng/ml human recombinant EGF, and 5.5 mM D-glucose. , culture was performed in a 37°C incubator in the presence of 5% CO 2 using medium supplemented with 750 ng/ml puromycin. Human pancreatic adenocarcinoma cell line MIAPACA2, human breast cancer cell line MDAMB231, and human hepatocellular carcinoma cell line HEP3B were purchased from KCLB and cultured in the presence of 5% CO 2 in an incubator at 37°C using DMEM/high glucose culture medium containing 10% FBS. Human prostate cancer cell line PC3 was purchased from KCLB and cultured in the presence of 5% CO 2 in an incubator at 37°C using RPMI culture medium containing 10% FBS.
인간 췌장 내피세포인 HPNE, 인간 신장암 세포주 (A498), 인간 췌장암 세포주 MIAPACA2, 인간 유방암 세포주 MDAMB231, 인간 간세포암 세포주 HEP3B, 인간 전립선암 세포주 PC3에 대한 PRD-2001의 효과를 확인하기 위해 Cell viability assay인 SRB assay를 진행하였다. 96-웰 플레이트에 세포를 배양하고, PRD-2001을 0, 1, 5 μM 농도로 처리하여 3일간 배양하였다. 50% TCA 용액을 50 ㎕ 추가하여 4℃에서 세포를 고정하였고 DW를 이용하여 5번 세척한 다음 플레이트를 건조하였다. 건조된 플레이트에 SRB 용액을 100 ㎕씩 상온에서 5분간 처리하고, 1% 아세트산을 이용하여 3번 세척한 다음, 상온에서 건조하였다. 건조된 플레이트에 100 ㎕ 10mM Tris 용액을 추가하여 30 분간 진탕 (shaking)하여 SRB를 녹이고 515 nm에서 흡광도를 측정하였다. Cell viability assay was performed to determine the effect of PRD-2001 on human pancreatic endothelial cells HPNE, human kidney cancer cell line (A498), human pancreatic cancer cell line MIAPACA2, human breast cancer cell line MDAMB231, human hepatocellular carcinoma cell line HEP3B, and human prostate cancer cell line PC3. SRB assay was performed. Cells were cultured in a 96-well plate, treated with PRD-2001 at concentrations of 0, 1, and 5 μM, and cultured for 3 days. Cells were fixed at 4°C by adding 50 ㎕ of 50% TCA solution, washed 5 times using DW, and the plate was dried. The dried plate was treated with 100 ㎕ of SRB solution at room temperature for 5 minutes, washed three times with 1% acetic acid, and then dried at room temperature. 100 μl of 10mM Tris solution was added to the dried plate and shaken for 30 minutes to dissolve SRB, and the absorbance was measured at 515 nm.
PRD-2001 처리에 따른 암세포 성장 억제율(대조군 대비 %)Cancer cell growth inhibition rate by PRD-2001 treatment (% compared to control group)
PRD-2001PRD-2001 HPNEHPNE A498A498 MIAPACA2MIAPACA2 MDAMB231 MDAMB231 HEP3BHEP3B PC3PC3
0 μM0 μM 100.0000100.0000 100.0000100.0000 100.0000100.0000 100.0000100.0000 100.0000100.0000 100.0000100.0000
1 μM1 μM 98.905298.9052 95.606195.6061 97.893697.8936 103.7414103.7414 93.605993.6059 91.412391.4123
5 μM5 μM 98.819098.8190 95.478695.4786 56.121356.1213 67.399767.3997 61.283061.2830 48.801348.8013
그 결과, 췌장암 세포주, 유방암 세포주, 간암 세포주 및 전립선암 세포주만 성장이 억제되었으며, 정상 췌장상피 세포주 및 신장암 세포주에 대해서는 성장 억제 효과가 없는 것을 확인하였다 [도 1]. 상기 [표 1]은 [도 1]의 암 세포주에 대한 성장 억제정도 (대조군 대비 %)를 나타낸다.As a result, it was confirmed that the growth of only pancreatic cancer cell lines, breast cancer cell lines, liver cancer cell lines, and prostate cancer cell lines was inhibited, and there was no growth inhibition effect on normal pancreatic epithelial cell lines and renal cancer cell lines [Figure 1]. [Table 1] shows the degree of growth inhibition (% compared to control group) for the cancer cell lines in [Figure 1].
실시예 2: PRD-2001에 의한 항암 효과 작용 기전 확인Example 2: Confirmation of anticancer effect mechanism by PRD-2001
본 발명에서는 PRD-2001에 의한 암세포 사멸 작용 기전을 확인하기 위해, 췌장암 세포주에 PRD-2001을 처리하였을 때, 췌장암 세포의 전체 전사체 분석을 수행하였다.In the present invention, in order to confirm the mechanism of cancer cell death by PRD-2001, when pancreatic cancer cell lines were treated with PRD-2001, the entire transcriptome analysis of pancreatic cancer cells was performed.
먼저, 인간 췌관선암종 세포주 BXPC3에서 PRD-2001 (10 μM)을 처리하여, 10일 배양한 다음 살아남은 세포에 대해, TRIzol을 이용해 전체 RNA를 추출한 후 마크로젠에 의뢰하여 RNA 염서열분석 (RNA sequencing)을 수행하였다. Bioanalyzer 2100과 RNA 6000 Nano Labchips를 사용하여 RNA의 품질과 양을 평가하였으며 Truseq RNA-Seq Library Prep Kit-v2를 이용해 RNA 라이브러리(RNA library)를 준비하였다. First, the human pancreatic adenocarcinoma cell line BXPC3 was treated with PRD-2001 (10 μM), cultured for 10 days, and total RNA was extracted from the surviving cells using TRIzol and then requested to Macrogen for RNA sequencing. carried out. The quality and quantity of RNA were evaluated using Bioanalyzer 2100 and RNA 6000 Nano Labchips, and an RNA library was prepared using Truseq RNA-Seq Library Prep Kit-v2.
이어 NovaSeq6000 sequencing system을 사용하여 paired-end sequencing을 수행하였다. 차등 발현 유전자 (Differentially expressed genes; DEGs)는 TPM (transcript per million mapped reads) 값이 10 미만인 값을 제외하고 선별하였다. PRD-2001에 의해 변형된 생물학적 기능을 연구하기 위해 GSEA(Gene Set Enrichment Analysis) 소프트웨어 v4.2.3을 사용하여 GSEA를 수행하였으며, GSEA 분석에 사용된 유전자 세트는 Reactome gene set 및 문헌 자료 (Ting Xi et al., PLoS One. 13(3): e0193427, 2018)로부터 수득하였다. 또한, 선별된 DEGs으로 IPA (Ingenuity pathway analysis)를 수행하여 Ingenuity Knowledge Base에 기반하는 주요 생물학적 기능을 식별하였다. Then, paired-end sequencing was performed using the NovaSeq6000 sequencing system. Differentially expressed genes (DEGs) were selected, excluding those with a TPM (transcript per million mapped reads) value of less than 10. To study the biological functions modified by PRD-2001, GSEA was performed using Gene Set Enrichment Analysis (GSEA) software v4.2.3, and the gene set used for GSEA analysis was the Reactome gene set and literature data (Ting Xi et al. al., PLoS One 13(3): e0193427, 2018). In addition, IPA (Ingenuity pathway analysis) was performed with the selected DEGs to identify key biological functions based on the Ingenuity Knowledge Base.
인간 췌관선암종 세포주 BXPC3는 KCLB에서 구매하였으며, 10% FBS를 포함하는 RPMI 배양 배지를 이용하여 37℃ 인큐베이터에서 5% CO2 존재 하에 배양하였다.The human pancreatic adenocarcinoma cell line BXPC3 was purchased from KCLB and cultured in the presence of 5% CO 2 in an incubator at 37°C using RPMI culture medium containing 10% FBS.
GSEA 분석 결과, 도 2에 나타난 바와 같이, PRD-2001 처리시 대조군에 비해, 췌장암 세포에서는 미분화 유전자군 [도 2A], 암세포의 성장 및 분열 과정인 세포주기 진행에 관여하는 유전자군 [도 2B]이 하향조절 되어 있음을 확인하였다. 또한, PRD-2001 처리 췌장암 세포에서 하향 조절된 유전자군은 암세포의 성장 및 DNA 손상 복구 기전에 작용하는 유전자임을 IPA 분석 [도 2C]을 통해 확인하였다.As a result of GSEA analysis, as shown in Figure 2, compared to the control group when treated with PRD-2001, pancreatic cancer cells showed an undifferentiated gene group [Figure 2A] and a gene group involved in cell cycle progression, which is the growth and division process of cancer cells [Figure 2B]. It was confirmed that this was downregulated. In addition, it was confirmed through IPA analysis [Figure 2C] that the gene group downregulated in PRD-2001-treated pancreatic cancer cells was a gene that played a role in cancer cell growth and DNA damage repair mechanisms.
실시예 3: PRD-2001 및 이리노테칸 병용투여에 따른 항암 시너지 효과 확인Example 3: Confirmation of anticancer synergy effect due to combined administration of PRD-2001 and irinotecan
본 발명에서는 PRD-2001과 항암제와의 병용투여에 따른 항암 시너지 효과를 확인하기 위해, 췌장암 세포주인 PANC1 및 BXPC3에 PRD-2001 및 이리노테칸을 단독 또는 병용투여한 후, 암세포 성장 정도를 콜로니 어세이 (Clonogenic assay)로 확인하였다. In the present invention, in order to confirm the anticancer synergy effect of the combined administration of PRD-2001 and anticancer drugs, PRD-2001 and irinotecan were administered alone or in combination to pancreatic cancer cell lines PANC1 and BXPC3, and then the degree of cancer cell growth was measured using a colony assay ( Clonogenic assay).
먼저, 6-웰 플레이트에 각각의 췌관선암종 세포주를 배양하고, PRD-2001은 0, 10 μM 농도가 되도록, 이리노테칸은 0, 0.5 μM 농도가 되도록 단독 또는 병용 처리하여 10일간 배양하였으며, 배양 동안 2 ~ 3일 마다 상기와 같은 조건의 배지로 교환해주었다. 그 다음, 0.02% 크리스탈 바이올렛 (Cristal Violet) 용액을 이용하여 세포를 고정 및 염색한 후, PBS를 이용하여 3번 세척한 다음, 플레이트를 건조시켰다. 건조된 플레이트는 이미징 후 'Image J' 프로그램의 Colony area 플러그인을 이용하여 정량하여 결과값을 확인하였다. 해당 결과값은 'Combenefit' 프로그램을 이용하여 분석을 진행하였으며, HSA 모델을 이용하여 시너지 이펙트 스코어 (synergy effect score)를 확인하였다. 시너지 이펙트 스코어 20 이상을 시너지 효과가 있는 것으로 판단하였다. First, each pancreatic adenocarcinoma cell line was cultured in a 6-well plate, and treated alone or in combination with PRD-2001 at a concentration of 0 and 10 μM and irinotecan at a concentration of 0 and 0.5 μM, and cultured for 10 days. During the culture, 2 The medium was replaced with a medium under the same conditions as above every ~3 days. Next, the cells were fixed and stained using 0.02% Crystal Violet solution, washed three times using PBS, and the plate was dried. The dried plate was imaged and then quantified using the Colony area plug-in of the 'Image J' program to confirm the results. The results were analyzed using the 'Combenefit' program, and the synergy effect score was confirmed using the HSA model. A synergy effect score of 20 or higher was judged to have a synergistic effect.
PRD-2001 및 이리노테칸 병용투여에 따른 암세포 성장 억제율(대조군 대비 %)Cancer cell growth inhibition rate (% compared to control group) following combined administration of PRD-2001 and irinotecan
PANC1PANC1 BXPC3BXPC3
ControlControl 100.0000100.0000 100.0000100.0000
PRD-2001 10 μMPRD-2001 10 μM 39.098139.0981 61.075161.0751
Irinotecan 0.5 μMIrinotecan 0.5 μM 36.292936.2929 73.527473.5274
PRD-2001 10 μM +
Irinotecan 0.5 μMPRD-2001 10 μM+
Irinotecan 0.5 μM 		6.11916.1191 11.483711.4837
그 결과, 도 3에 나타난 바와 같이, PANC1 및 BxPC3 모두에서 단독투여군에 비해 PRD-2001 및 이리노테칸 병용투여군에서 암세포 성장이 현저하게 억제되는 것으로 나타났다. 상기 [표 2]는 [도 3]의 암 세포주에 대한 성장 억제정도 (대조군 대비 %)를 나타낸다.또한, PANC1 및 BxPC3 각각에 대한 병용투여에 의한 시너지 점수(Synergy score)는 30 및 50으로, 병용투여에 따른 항암 시너지 효과를 확인하였다. As a result, as shown in Figure 3, cancer cell growth was found to be significantly inhibited in the PRD-2001 and irinotecan combination group compared to the single group in both PANC1 and BxPC3. [Table 2] shows the degree of growth inhibition (% compared to control group) for the cancer cell lines in [Figure 3]. In addition, the synergy score by combined administration for PANC1 and BxPC3, respectively, was 30 and 50, The anticancer synergy effect of combined administration was confirmed.
실시예 4: 동물모델에서 PRD-2001 투여에 따른 항암 효과 확인Example 4: Confirmation of anticancer effect following PRD-2001 administration in animal model
본 발명에서는 생체 내에서 PRD-2001 투여에 따른 항암 효과를 확인하기 위해, 췌장암 이종이식 종양 모델을 제작하여 약물을 투여하였다. In the present invention, in order to confirm the anticancer effect of PRD-2001 administration in vivo, a pancreatic cancer xenograft tumor model was created and the drug was administered.
먼저, 6-8주령 Balb/c 누드 마우스에 100 ㎕ PBS 중 MIAPACA2 세포(1 × 107)를 1 ㎖ 시린지를 사용하여 피하로 접종하였으며, 종양의 크기가 100 ㎣에 도달했을 때, 마우스를 하기와 같이 그룹당 5 마리가 되도록 3개의 그룹으로 무작위로 나눈 다음, PRD-2001을 투여하였다. PRD-2001은 5% DMSO와 3% Cremophor EL 와 92% 생리식염수에 녹였다. First, 6-8 week old Balb/c nude mice were subcutaneously inoculated with MIAPACA2 cells ( 1 The animals were randomly divided into 3 groups with 5 animals per group as shown, and then PRD-2001 was administered. PRD-2001 was dissolved in 5% DMSO, 3% Cremophor EL, and 92% saline solution.
1) 대조군(용매처리)1) Control group (solvent treatment)
2) PRD-2001 단독 처리군 (50 mg/kg I.P injection, 주 5회)2) PRD-2001 treatment group alone (50 mg/kg I.P injection, 5 times a week)
3) PRD-2001 단독 처리군 (100 mg/kg I.P injection, 주 5회)3) PRD-2001 treatment group alone (100 mg/kg I.P injection, 5 times a week)
종양 크기는 캘리퍼(Caliper)를 이용하여 주 1회 측정하였고, 마우스의 체중 또한 주 1회 측정하였다. 종양의 크기는 하기 수학식 1을 이용하여 계산하였다. Tumor size was measured once a week using a caliper, and the weight of the mouse was also measured once a week. The size of the tumor was calculated using Equation 1 below.
[수학식 1][Equation 1]
종양 크기 = (단축 x 단축 x 장축)/2Tumor size = (short axis x short axis x long axis)/2
본 실험은 국립 암센터 연구소의 기관 동물 관리 및 사용위원회 (IACUC)에 의해 검토 및 승인되었다 (IACUC 승인번호; NCC-22-796).This experiment was reviewed and approved by the Institutional Animal Care and Use Committee (IACUC) of the National Cancer Center Research Institute (IACUC approval number; NCC-22-796).
PRD-2001 투여에 따른 종양 부피Tumor volume according to PRD-2001 administration
Tumor volume (mm3)Tumor volume ( mm3 ) 0 day0 day 7 day7 days 14 day14 days 21 day21 days
ControlControl 72.801672.8016 148.4274148.4274 255.9562255.9562 388.6936388.6936
PRD-2001 50 mg/kgPRD-2001 50mg/kg 55.542655.5426 122.5912122.5912 208.0494208.0494 344.134344.134
PRD-2001 100 mg/kgPRD-2001 100 mg/kg 80.123680.1236 108.784108.784 96.022896.0228 181.6902181.6902
PRD-2001 투여에 따른 최종 종양 무게Final tumor weight according to PRD-2001 administration
Tumor weight (g)Tumor weight (g)
ControlControl 0.31800.3180
PRD-2001 50 mg/kgPRD-2001 50mg/kg 0.27200.2720
PRD-2001 100 mg/kgPRD-2001 100 mg/kg 0.12800.1280
그 결과, [도 4A] ~ [도 4C]에 나타난 바와 같이, 암세포 이종이식 종양 모델에 PRD-2001을 투여한 결과, 대조군 비해 PRD-2001의 농도에 따라 종양조직 크기가 현저하게 감소하는 것을 확인하였다. 상기 [표 3]와 [표 4]는 [도 4]의 종양동물모델에서 시간에 따른 종양의 부피와 최종 종양의 무게를 나타낸다.As a result, as shown in [Figure 4A] to [Figure 4C], when PRD-2001 was administered to a cancer cell xenograft tumor model, it was confirmed that the size of tumor tissue was significantly reduced depending on the concentration of PRD-2001 compared to the control group. did. [Table 3] and [Table 4] show the tumor volume and final tumor weight over time in the tumor animal model of [Figure 4].
실시예 5: 동물모델에서 PRD-2001 및 이리노테칸 병용투여에 따른 항암 시너지 효과 확인Example 5: Confirmation of anticancer synergy effect due to combined administration of PRD-2001 and irinotecan in animal model
본 발명에서는 생체 내에서 PRD-2001 및 이리노테칸 병용투여에 따른 항암 시너지 효과를 확인하기 위해, 췌장암 이종이식 종양 모델을 제작하여 약물을 투여하였다. In the present invention, in order to confirm the anticancer synergistic effect of the combined administration of PRD-2001 and irinotecan in vivo, a pancreatic cancer xenograft tumor model was created and the drug was administered.
먼저, 6-8주령 Balb/c 누드 마우스에 100 ㎕ PBS 중 MIA PaCa-2 세포(1 × 107)를 1 ㎖ 시린지를 사용하여 피하로 접종하였으며, 종양의 크기가 100 ㎣에 도달했을 때, 마우스를 하기와 같이 그룹당 3 마리가 되도록 4개의 그룹으로 무작위로 나눈 다음, PRD-2001 및 이리노테칸을 투여하였다. PRD-2001은 10% DMSO와 90% 생리식염수에 녹여 준비하였으며 이리노테칸은 4% DMSO와 96% 생리식염수에 녹여 준비하였다. First, 6-8 week old Balb/c nude mice were subcutaneously inoculated with MIA PaCa-2 cells ( 1 Mice were randomly divided into 4 groups with 3 mice per group as follows, and then PRD-2001 and irinotecan were administered. PRD-2001 was prepared by dissolving in 10% DMSO and 90% saline, and irinotecan was prepared by dissolving in 4% DMSO and 96% saline.
1) 대조군(용매처리)1) Control group (solvent treatment)
2) PRD-2001 단독 처리군 (50 mg/kg I.P injection, 주 5회),2) PRD-2001 treatment group alone (50 mg/kg I.P injection, 5 times a week),
3) 이리노테칸 단독 처리군 (20 mg/kg, I.P injection, 주 1회),3) Irinotecan treatment group alone (20 mg/kg, I.P injection, once a week),
4) PRD-2001 (50 mg/kg I.P injection, 주 5회) 및 이리노테칸 (20 mg/kg, I.P injection, 주 1회) 병용 처리군4) PRD-2001 (50 mg/kg I.P injection, 5 times a week) and irinotecan (20 mg/kg, I.P injection, once a week) combination treatment group
종양 크기는 캘리퍼(Caliper)를 이용하여 주 2회 측정하였고, 마우스의 체중 또한 주 2회 측정하였다. 종양의 크기는 하기 수학식 1을 이용하여 계산하였다. Tumor size was measured twice a week using a caliper, and the body weight of the mouse was also measured twice a week. The size of the tumor was calculated using Equation 1 below.
[수학식 1][Equation 1]
종양 크기 = (단축 x 단축 x 장축)/2Tumor size = (short axis x short axis x long axis)/2
본 실험은 국립 암센터 연구소의 기관 동물 관리 및 사용위원회 (IACUC)에 의해 검토 및 승인되었다 (IACUC 승인번호; NCC-20-579).This experiment was reviewed and approved by the Institutional Animal Care and Use Committee (IACUC) of the National Cancer Center Research Institute (IACUC approval number; NCC-20-579).
PRD-2001 및 이리노테칸 병용 투여에 따른 종양 부피Tumor volume following PRD-2001 and irinotecan co-administration
Tumor volume (mm3)Tumor volume ( mm3 ) 0 day0 days 5 day5 days 8 day8 days 12 day12 days 15 day15 days
ControlControl 155.6883155.6883 258.2663258.2663 377.2575377.2575 648.5885648.5885 842.448842.448
PRD-2001 50 mg/kgPRD-2001 50mg/kg 120.3473120.3473 214.1228214.1228 283.558283.558 621.017621.017 823.444823.444
Irinotecan 20 mg/kg Irinotecan 20 mg/kg 108.0405108.0405 121.77121.77 159.719159.719 301.077301.077 501.7455501.7455
PRD-2001 50 mg/kg + Irinotecan 20 mg/kgPRD-2001 50 mg/kg + Irinotecan 20 mg/kg 78.9997578.99975 118.8765118.8765 134.514134.514 176.176176.176 149.2553149.2553
PRD-2001 및 이리노테칸 병용 투여에 따른 최종 종양 무게Final tumor weight following combined administration of PRD-2001 and irinotecan
Tumor weight (g)Tumor weight (g)
ControlControl 0.66950.6695
PRD-2001 50 mg/kgPRD-2001 50mg/kg 0.3520.352
Irinotecan 20 mg/kg Irinotecan 20 mg/kg 0.5670.567
PRD-2001 50 mg/kg + Irinotecan 20 mg/kgPRD-2001 50 mg/kg + Irinotecan 20 mg/kg 0.0720.072
PRD-2001 및 이리노테칸 병용 투여에 따른 마우스 체중Mouse body weight according to combined administration of PRD-2001 and irinotecan
Mouse weight (g)Mouse weight (g) 0 day0 days 5 day5 days 8 day8 days 12 day12 days 15 day15 days
ControlControl 18.318.3 18.6518.65 19.619.6 20.220.2 20.220.2
PRD-2001 50 mg/kgPRD-2001 50mg/kg 18.0518.05 18.1518.15 18.818.8 19.519.5 19.5519.55
Irinotecan 20 mg/kg Irinotecan 20 mg/kg 1818 17.417.4 18.4518.45 17.9517.95 18.6518.65
PRD-2001 50 mg/kg + Irinotecan 20 mg/kgPRD-2001 50 mg/kg + Irinotecan 20 mg/kg 18.218.2 17.917.9 18.3518.35 18.718.7 19.3519.35
그 결과, [도 5A] ~ [도 5B]에 나타난 바와 같이, 암세포 이종이식 종양 모델에 PRD-2001 및 이리노테칸을 병용투여한 결과, 대조군 및 단독투여군에 비해 종양조직 크기가 현저하게 감소하는 것을 확인하였으며, 약물 투여에 따른 마우스 체중 감소는 관찰되지 않았으므로 [도 5C], 종양 성장 및 암세포 성장 억제는 약물의 부작용이 아닌 두 약물의 시너지 효과임을 확인하였다. 상기 [표 5] 내지 [표 6]은 [도 5]의 종양동물모델에서 시간에 따른 종양의 부피, 종양의 최종 무게, 마우스의 몸무게 변화를 나타낸다.As a result, as shown in [Figure 5A] to [Figure 5B], when PRD-2001 and irinotecan were administered together in a cancer cell xenograft tumor model, it was confirmed that the tumor tissue size was significantly reduced compared to the control group and the single administration group. Since no decrease in mouse weight was observed following drug administration [Figure 5C], it was confirmed that inhibition of tumor growth and cancer cell growth was not a side effect of the drug but a synergistic effect of the two drugs. [Table 5] to [Table 6] show changes in tumor volume, final tumor weight, and mouse body weight over time in the tumor animal model shown in [FIG. 5].
실시예 6: PRD-2001 및 항암제 병용투여에 따른 항암 시너지 효과 확인Example 6: Confirmation of anticancer synergy effect due to combined administration of PRD-2001 and anticancer drugs
본 발명에서는 PRD-2001과 다양한 항암제와의 병용투여시 항암 시너지 효과를 보이는지 확인하기 위해, 상기 실시예 3과 동일한 방법으로 플루오로우라실(fluorouracil, 5-FU, 1 μM), 파클리탁셀(Paclitaxel, 1.25 nM), 독소루비신(Doxorubicin, 5 nM), 또는 젬시타빈(Gemcitabine, 2.5 nM)을 PRD-2001(10 μM)과 단독 또는 병용투여하였다. In the present invention, in order to confirm whether PRD-2001 shows an anticancer synergy effect when administered in combination with various anticancer drugs, fluorouracil (5-FU, 1 μM) and paclitaxel (Paclitaxel, 1.25%) were used in the same manner as Example 3. nM), Doxorubicin (5 nM), or Gemcitabine (2.5 nM) were administered alone or in combination with PRD-2001 (10 μM).
PRD-2001 및 젬시타빈 병용처리에 따른 암세포 성장 억제율(대조군 대비 %)Cancer cell growth inhibition rate following PRD-2001 and gemcitabine combination treatment (% compared to control group)
MIAPACA2MIAPACA2
ControlControl 100.0000100.0000
PRD-2001 10 μMPRD-2001 10 μM 91.423091.4230
Gemcitabine 2.5 nMGemcitabine 2.5 nM 60.331460.3314
PRD-2001 10 μM + Gemcitabine 2.5 nMPRD-2001 10 μM + Gemcitabine 2.5 nM 35.900935.9009
PRD-2001 및 독소루비신 병용처리에 따른 암세포 성장 억제율(대조군 대비 %)Cancer cell growth inhibition rate following PRD-2001 and doxorubicin combination treatment (% compared to control group)
MIAPACA2MIAPACA2
ControlControl 100.0000100.0000
PRD-2001 10 μMPRD-2001 10 μM 70.473670.4736
Doxorubicin 5 nM Doxorubicin 5 nM 54.746354.7463
PRD-2001 10 μM + Doxorubicin 5 nMPRD-2001 10 μM + Doxorubicin 5 nM 31.276231.2762
PRD-2001 및 파클리탁셀 병용처리에 따른 암세포 성장 억제율(대조군 대비 %)Cancer cell growth inhibition rate following PRD-2001 and paclitaxel combination treatment (% compared to control group)
MIAPACA2MIAPACA2
ControlControl 100.0000100.0000
PRD-2001 10 μMPRD-2001 10 μM 43.130643.1306
Paclitaxel 1.25 nMPaclitaxel 1.25 nM 38.818238.8182
PRD-2001 10 μM + Paclitaxel 1.25 nMPRD-2001 10 μM + Paclitaxel 1.25 nM 18.359518.3595
PRD-2001 및 5-FU 병용처리에 따른 암세포 성장 억제율(대조군 대비 %)Cancer cell growth inhibition rate by PRD-2001 and 5-FU combination treatment (% compared to control group)
MIAPACA2MIAPACA2
ControlControl 100.0000100.0000
PRD-2001 10 μMPRD-2001 10 μM 86.258586.2585
5FU 1 μM 5FU 1 μM 59.337759.3377
PRD-2001 10 μM + 5FU 1 μMPRD-2001 10 μM + 5FU 1 μM 47.228047.2280
그 결과, 도 6에 나타난 바와 같이, 젬시타빈, 독소루비신 및 파클리탁셀은 PRD-2001과 병용투여에 따른 항암 시너지 효과를 보이는 것으로 나타났으며, 5-FU은 PRD-2001과의 병용투여에 대한 시너지 효과를 보이지 않는 것을 확인하였다. 상기 [표 8] 내지 [표 11]은 [도 6]의 암 세포주에 대한 성장 억제정도 (대조군 대비 %)를 나타낸다.As a result, as shown in Figure 6, gemcitabine, doxorubicin, and paclitaxel were shown to have an anticancer synergy effect when administered in combination with PRD-2001, and 5-FU showed a synergistic effect when administered in combination with PRD-2001. It was confirmed that was not visible. [Table 8] to [Table 11] show the degree of growth inhibition (% compared to control group) for the cancer cell line of [Figure 6].
본 발명에서는 PRD-2001에 의해 췌장암, 유방암, 전립선암, 간암 세포의 성장을 억제하는 것을 확인하였으며, PRD-2001과 이리노테칸 (Irinotecan), 젬시타빈 (gemcitabine), 파클리탁셀 (Paclitaxel) 또는 독소루비신 (Doxorubicin)의 병용투여에 따른 항암 시너지 효과를 확인하였으므로, 본 발명의 조성물은 효과적인 항암제로 유용하게 활용할 수 있다. In the present invention, it was confirmed that PRD-2001 inhibits the growth of pancreatic cancer, breast cancer, prostate cancer, and liver cancer cells, and PRD-2001 and irinotecan, gemcitabine, paclitaxel, or doxorubicin Since the anti-cancer synergy effect of combined administration was confirmed, the composition of the present invention can be usefully used as an effective anti-cancer agent.

Claims (10)

  1. 하기 화학식 1로 표시되는 PRD-2001 (우르손산; ursonic acid) 또는 이의 약제학적으로 허용가능한 염을 유효성분으로 포함하는, 췌장암, 유방암, 간암 또는 전립선암 예방 또는 치료용 약학적 조성물:A pharmaceutical composition for preventing or treating pancreatic cancer, breast cancer, liver cancer or prostate cancer, comprising PRD-2001 (ursonic acid) or a pharmaceutically acceptable salt thereof represented by the following formula (1) as an active ingredient:
    [화학식 1][Formula 1]
    Figure PCTKR2023016368-appb-img-000005
    .
    Figure PCTKR2023016368-appb-img-000005
    .
  2. 제1항에 있어서, According to paragraph 1,
    상기 PRD-2001은 미분화 암세포 분화 유도, 암세포 성장 억제 또는 암세포 DNA 손상 복구를 억제하는 것을 특징으로 하는, 췌장암, 유방암, 간암 또는 전립선암 예방 또는 치료용 약학적 조성물.The PRD-2001 is a pharmaceutical composition for preventing or treating pancreatic cancer, breast cancer, liver cancer, or prostate cancer, characterized in that it induces differentiation of undifferentiated cancer cells, inhibits cancer cell growth, or inhibits DNA damage repair in cancer cells.
  3. 하기 화학식 1로 표시되는 PRD-2001 (우르손산; ursonic acid) 또는 이의 약제학적으로 허용가능한 염을 유효성분으로 포함하는, 췌장암, 유방암, 간암 또는 전립선암에 대한 항암 보조제:Anti-cancer adjuvant for pancreatic cancer, breast cancer, liver cancer or prostate cancer, comprising PRD-2001 (ursonic acid) or a pharmaceutically acceptable salt thereof represented by the following formula (1) as an active ingredient:
    [화학식 1][Formula 1]
    Figure PCTKR2023016368-appb-img-000006
    .
    Figure PCTKR2023016368-appb-img-000006
    .
  4. 제3항에 있어서, According to paragraph 3,
    상기 PRD-2001은 미분화 암세포 분화 유도, 암세포 성장 억제 또는 암세포 DNA 손상 복구를 억제하는 것을 특징으로 하는, 췌장암, 유방암, 간암 또는 전립선암에 대한 항암 보조제.The PRD-2001 is an anticancer adjuvant for pancreatic cancer, breast cancer, liver cancer, or prostate cancer, characterized in that it induces differentiation of undifferentiated cancer cells, inhibits cancer cell growth, or inhibits DNA damage repair in cancer cells.
  5. 하기 화학식 1로 표시되는 PRD-2001 (우르손산; ursonic acid) 또는 이의 약제학적으로 허용가능한 염; 및 PRD-2001 (ursonic acid) represented by the following formula (1) or a pharmaceutically acceptable salt thereof; and
    [화학식 1][Formula 1]
    Figure PCTKR2023016368-appb-img-000007
    .
    Figure PCTKR2023016368-appb-img-000007
    .
    이리노테칸 (Irinotecan), 젬시타빈 (gemcitabine), 파클리탁셀 (Paclitaxel) 및 독소루비신 (Doxorubicin)으로 구성된 군에서 선택되는 어느 하나 이상의 항암제를 유효성분으로 포함하는, 암 예방 또는 치료용 약학적 조성물.A pharmaceutical composition for preventing or treating cancer, comprising as an active ingredient one or more anticancer agents selected from the group consisting of Irinotecan, gemcitabine, Paclitaxel, and Doxorubicin.
  6. 제5항에 있어서, According to clause 5,
    상기 PRD-2001은 미분화 암세포 분화 유도, 암세포 성장 억제 또는 암세포 DNA 손상 복구를 억제하는 것을 특징으로 하는, 암 예방 또는 치료용 약학적 조성물.The PRD-2001 is a pharmaceutical composition for preventing or treating cancer, characterized in that it induces differentiation of undifferentiated cancer cells, inhibits cancer cell growth, or inhibits DNA damage repair in cancer cells.
  7. 제5항에 있어서, According to clause 5,
    상기 암은 췌장암, 유방암, 간암, 전립선암, 대장암, 폐암, 위암, 흑색종, 전립선암, 난소암, 및 교모세포종 으로 이루어지는 군에서 선택되는 어느 하나 이상의 암인 것을 특징으로 하는, 암 예방 또는 치료용 약학적 조성물.Cancer prevention or treatment, characterized in that the cancer is any one or more cancers selected from the group consisting of pancreatic cancer, breast cancer, liver cancer, prostate cancer, colon cancer, lung cancer, stomach cancer, melanoma, prostate cancer, ovarian cancer, and glioblastoma. Pharmaceutical composition.
  8. 하기 화학식 1로 표시되는 PRD-2001 (우르손산; ursonic acid) 또는 이의 약제학적으로 허용가능한 염; 및 PRD-2001 (ursonic acid) represented by the following formula (1) or a pharmaceutically acceptable salt thereof; and
    [화학식 1][Formula 1]
    Figure PCTKR2023016368-appb-img-000008
    Figure PCTKR2023016368-appb-img-000008
    이리노테칸 (Irinotecan), 젬시타빈 (gemcitabine), 파클리탁셀 (Paclitaxel) 및 독소루비신 (Doxorubicin)으로 구성된 군에서 선택되는 어느 하나 이상의 항암제를 유효성분으로 포함하는, 항암 보조제.An anticancer adjuvant containing as an active ingredient one or more anticancer agents selected from the group consisting of Irinotecan, gemcitabine, Paclitaxel, and Doxorubicin.
  9. 제8항에 있어서, According to clause 8,
    상기 PRD-2001은 미분화 암세포 분화 유도, 암세포 성장 억제 또는 암세포 DNA 손상 복구를 억제하는 것을 특징으로 하는, 항암 보조제.The PRD-2001 is an anti-cancer adjuvant characterized in that it induces differentiation of undifferentiated cancer cells, inhibits cancer cell growth, or inhibits DNA damage repair in cancer cells.
  10. 제8항에 있어서, According to clause 8,
    상기 암은 췌장암, 유방암, 간암, 전립선암, 대장암, 폐암, 위암, 흑색종, 전립선암, 난소암, 및 교모세포종 으로 이루어지는 군에서 선택되는 어느 하나 이상의 암인 것을 특징으로 하는, 항암 보조제.An anti-cancer adjuvant, wherein the cancer is one or more cancers selected from the group consisting of pancreatic cancer, breast cancer, liver cancer, prostate cancer, colon cancer, lung cancer, stomach cancer, melanoma, prostate cancer, ovarian cancer, and glioblastoma.
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