WO2021112541A1 - Pharmaceutical composition for prevention and treatment of secondary cancer, comprising aldehyde inhibitor and biguanide-based compound - Google Patents

Pharmaceutical composition for prevention and treatment of secondary cancer, comprising aldehyde inhibitor and biguanide-based compound Download PDF

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WO2021112541A1
WO2021112541A1 PCT/KR2020/017417 KR2020017417W WO2021112541A1 WO 2021112541 A1 WO2021112541 A1 WO 2021112541A1 KR 2020017417 W KR2020017417 W KR 2020017417W WO 2021112541 A1 WO2021112541 A1 WO 2021112541A1
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cancer
pharmaceutical composition
biguanide
based compound
phenformin
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PCT/KR2020/017417
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French (fr)
Korean (ko)
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김홍렬
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주식회사 하임바이오
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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
    • A61K31/12—Ketones
    • 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
    • A61K31/13—Amines
    • A61K31/155—Amidines (), e.g. guanidine (H2N—C(=NH)—NH2), isourea (N=C(OH)—NH2), isothiourea (—N=C(SH)—NH2)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2300/00—Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00

Definitions

  • the present invention relates to a pharmaceutical composition for the prevention and treatment of secondary cancer comprising an aldehyde inhibitor and a biguanide-based compound.
  • Colorectal cancer is a malignant tumor that occurs in the colon. According to a report by the International Agency for Research on Cancer (IARC) under the World Health Organization (WHO), the incidence rate of colorectal cancer in Korea is 45 per 100,000 people, the highest in the world. According to the cancer registration statistics announced by the Central Cancer Registry, there were 217,057 cancer cases in Korea in 2014, of which 26,978 cases were colorectal cancer, accounting for 12.4% of the total, taking the third place with a high incidence rate. Mortality due to colorectal cancer is also high enough to occupy the fourth place in the number of deaths by cancer type. The increase in the incidence of colorectal cancer is further accelerated by the increase in life expectancy and the westernized diet.
  • IARC International Agency for Research on Cancer
  • WHO World Health Organization
  • secondary cancer refers to a secondary cancer that has metastasized to a new organ by first metastasizing cancer from another site, and has a poorer prognosis than primary cancer.
  • the present invention has been devised to solve the above problems, and relates to a pharmaceutical composition for the prevention and treatment of secondary cancer comprising an aldehyde inhibitor and a biguanide-based compound. Since the pharmaceutical composition of the present invention has a significant therapeutic effect in secondary cancer specifically compared to primary cancer, it is expected to be widely used in the treatment of secondary cancer, which is difficult to treat and has a poor prognosis.
  • the present invention relates to a pharmaceutical composition for the prevention and treatment of secondary cancer comprising an aldehyde inhibitor and a biguanide-based compound.
  • cancer is characterized by uncontrolled cell growth, and by this abnormal cell growth, a cell mass called a tumor is formed, penetrates into surrounding tissues, and in severe cases, it spreads to other organs of the body. It is said to be transferable. Scientifically, it is also called a neoplasm. Cancer is an intractable chronic disease that in many cases cannot be cured fundamentally even if treated with surgery, radiation, and chemotherapy, but causes pain and ultimately death. There are several causes of cancer, but internal factors and external factors. It is not known exactly how normal cells are transformed into cancer cells, but it is known that a significant number of cancers are affected by external factors such as environmental factors. Internal factors include genetic factors and immunological factors, and external factors include chemicals, radiation, and viruses. The genes involved in the development of cancer include oncogenes and tumor suppressor genes, and cancer occurs when the balance between them is disrupted by the internal or external factors described above.
  • primary cancer refers to a cancer that originally occurred in situ. It also implies that it occurred first in the organ. In relative terms, it is distinguished from secondary cancer. For example, there is “primary colorectal cancer” in which cells that were originally in the colon have turned into cancer cells.
  • second cancer is a meaning distinct from the primary cancer, and refers to cancer as a result of other causes. For example, if cancer cells originating elsewhere metastasize to the colon and colonize and then grow cancerous tissue, it is called “secondary colorectal cancer”.
  • cancer metastasis means that cancer cells leave a primary organ and go to another organ, and the term “cancer” includes “cancer stem cells”.
  • cancer includes “cancer stem cells”.
  • the spread of cancer to other parts of the body is largely divided into growth of cancer tissue from the primary cancer and directly infiltrating surrounding organs, and distant metastasis along blood vessels or lymphatic vessels to other distant organs.
  • treatment refers to a series of activities performed for alleviation and/or amelioration of a desired disease.
  • treatment is to inhibit numerical or quantitative proliferation of cancer cells including cancer stem cells in secondary cancer, kill cancer cells, inhibit the growth of cancer tissues, or reduce the size of cancer tissues. or inhibiting the development of new blood vessels in cancer tissue, or inhibiting the invasion or metastasis of cancer to another site.
  • the present inventors have completed the present invention by confirming that, as a result of the study, when an aldehyde inhibitor and a biguanide-based compound are administered in combination, the cancer treatment effect specifically for secondary cancer is remarkable.
  • the present invention relates to a pharmaceutical composition for preventing or treating secondary cancer comprising an aldehyde inhibitor and a biguanide-based compound, preferably the aldehyde inhibitor is 3-hydroxykynurenine ( 3-hydroxy-DL-kynurenine; 3-HDK), benomyl, cis-diamminedichloridoplatinum (CDDP), chlorpropamide, citral, CVT-10216 (3-[[[ 3-[4-[(methylsulfonyl)amino]phenyl]-4-oxo-4H-1-benzopyran-7-yl]oxy]methyl]-benzoic acid, or 3-[[[3-[4-[(methylsulfonyl )amino]phenyl]-4-oxo-4H-chromen-7-yl]oxy]methyl]benzoic acid), cyanamide, daidzin, diethylaminobenzaldehyde (DEAB), diethylaminobenzaldehyde, die
  • the aldehyde inhibitor and the biguanide-based compound may be included in a weight ratio of 1:1 to 100, preferably in a weight ratio of 1:2 to 20.
  • the aldehyde inhibitor may be included in an amount of 0.5-50 ⁇ M.
  • the biguanide-based compound may be included in an amount of 10 to 1000 ⁇ M.
  • the composition of the present invention can prevent and/or treat secondary cancer by inhibiting it, preferably, it is characterized in that it can effectively prevent and/or treat secondary colorectal cancer.
  • the pharmaceutical composition of the present invention may be additionally administered in combination with other anticancer agents, and through this, it can be used to effectively treat cancer stem cells as well as general cancer cells.
  • the anticancer agent is nitrogen mustard, imatinib, oxaliplatin, rituximab, erlotinib, neratinib, lapatinib, gefitinib, vandetanib, nirotinib, semasanib, bosutinib, axitinib, cediranib , restautinib, trastuzumab, gefitinib, bortezomib, sunitinib, carboplatin, sorafenib, bevacizumab, cisplatin, cetuximab, viscumalbum, asparaginase, tretinoin, hydroxycarba amide, dasatinib, estramustine, gemtuzumab ozogamicin, ibritumomab tuccetan, heptaplatin, methylaminolevulinic acid, amsacrine, alemtuzum
  • the pharmaceutical composition may be characterized in the form of capsules, tablets, granules, injections, ointments, powders or beverages, and the pharmaceutical composition may be characterized in that it is intended for humans.
  • the pharmaceutical composition of the present invention is not limited thereto, but can be formulated and used in the form of oral dosage forms such as powders, granules, capsules, tablets, aqueous suspensions, external preparations, suppositories, and sterile injection solutions, respectively, according to conventional methods.
  • the pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier.
  • Pharmaceutically acceptable carriers may include binders, lubricants, disintegrants, excipients, solubilizers, dispersants, stabilizers, suspending agents, dyes, fragrances, etc., for oral administration, and in the case of injections, buffers, preservatives, pain-freezing agents A topical agent, solubilizer, isotonic agent, stabilizer, etc.
  • the dosage form of the pharmaceutical composition of the present invention can be prepared in various ways by mixing with a pharmaceutically acceptable carrier as described above.
  • a pharmaceutically acceptable carrier as described above.
  • it can be prepared in the form of tablets, troches, capsules, elixirs, suspensions, syrups, wafers, etc., and in the case of injections, it can be prepared in the form of unit dose ampoules or multiple doses. have.
  • it can be formulated as a solution, suspension, tablet, capsule, sustained release formulation, and the like.
  • suitable carriers, excipients and diluents for formulation include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, malditol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate or mineral oil may be used.
  • fillers, anti-agglomeration agents, lubricants, wetting agents, fragrances, emulsifiers, preservatives and the like may be further included.
  • the route of administration of the pharmaceutical composition according to the present invention is, but not limited to, oral, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, intestinal, topical, sublingual or rectal. Oral or parenteral administration is preferred.
  • parenteral includes subcutaneous, intradermal, intravenous, intramuscular, intraarticular, intrabursar, intrasternal, intrathecal, intralesional and intracranial injection or infusion techniques.
  • the pharmaceutical composition of the present invention may also be administered in the form of a suppository for rectal administration.
  • the pharmaceutical composition of the present invention varies depending on several factors including the activity of the specific compound used, age, weight, general health, sex, formula, administration time, administration route, excretion rate, drug formulation, and the severity of the specific disease to be prevented or treated
  • the dosage of the pharmaceutical composition may vary depending on the patient's condition, weight, degree of disease, drug form, administration route and period, but may be appropriately selected by those skilled in the art, and 0.0001 to 50 mg/kg per day or It may be administered at 0.001 to 50 mg/kg. Administration may be administered once a day, or may be administered in several divided doses. The above dosage does not limit the scope of the present invention in any way.
  • the pharmaceutical composition according to the present invention may be formulated as pills, dragees, capsules, solutions, gels, syrups, slurries, and suspensions.
  • "food composition” is used in various ways for the prevention or improvement of indications for the purpose of the present invention
  • the food composition comprising the composition of the present invention as an active ingredient includes various foods, such as For example, it may be prepared in the form of beverage, gum, tea, vitamin complex, powder, granule, tablet, capsule, confectionery, rice cake, bread, and the like.
  • the food composition of the present invention can be safely used even when taken for a long time for the purpose of preventing the indications of the present invention.
  • the amount may be added in a proportion of 0.1 to 100% of the total weight.
  • the food composition when the food composition is prepared in the form of a beverage, there is no particular limitation other than containing the food composition in the indicated ratio, and it may contain various flavoring agents or natural carbohydrates as additional ingredients, like a conventional beverage. That is, as natural carbohydrates, monosaccharides such as glucose, disaccharides such as fructose, and polysaccharides such as sucrose, common sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol are included. can do.
  • natural carbohydrates monosaccharides such as glucose, disaccharides such as fructose, and polysaccharides such as sucrose, common sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol are included. can do.
  • flavoring agent examples include natural flavoring agents (taumatin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.).
  • These components can be used independently or in combination.
  • the proportion of these additives is usually per 100 parts by weight of the composition of the present invention. It is generally selected in the range of 0.1 to 100 parts by weight, but is not limited thereto.
  • a pharmaceutical composition for preventing or treating secondary cancer comprising an aldehyde inhibitor and a biguanide-based compound, wherein the aldehyde inhibitor is Gossypol It provides a pharmaceutical composition comprising the above, wherein the biguanide-based compound is phenformin, wherein the aldehyde inhibitor and the biguanide-based compound are included in a weight ratio of 1:1 to 100.
  • the aldehyde inhibitor and the biguanide-based compound provides a pharmaceutical composition characterized in that it is included in a weight ratio of 1: 2 to 20, the aldehyde inhibitor is in an amount of 0.5 to 50 ⁇ M
  • the biguanide-based compound provides a pharmaceutical composition characterized in that it is included in an amount of 10 to 1000 ⁇ M, wherein the secondary cancer is secondary colorectal cancer. It provides a composition, wherein the secondary cancer has infiltrated from nearby organs or has spread remotely from blood vessels or lymph nodes, wherein the cancer comprises cancer stem cells. to provide.
  • a method for preventing or treating secondary cancer with a pharmaceutical composition comprising an aldehyde inhibitor and a biguanide-based compound as an active ingredient, wherein the aldehyde inhibitor is Gossypol (Gossypol) provides a method for preventing or treating secondary cancer, wherein the biguanide-based compound is phenformin (Phenformin).
  • Gossypol Gossypol
  • Phenformin phenformin
  • a secondary cancer prevention or treatment use of an aldehyde inhibitor and a biguanide-based compound wherein the aldehyde inhibitor is Gossypol secondary cancer It provides a preventive or therapeutic use, and the biguanide-based compound provides a secondary cancer prevention or treatment use, characterized in that the biguanide-based compound is phenformin.
  • a secondary cancer prevention or treatment use of a pharmaceutical composition comprising an aldehyde inhibitor and a biguanide-based compound as an active ingredient, wherein the aldehyde inhibitor is Gossypol ) provides a secondary cancer prevention or treatment use, characterized in that the biguanide-based compound provides a secondary cancer prevention or treatment use, characterized in that the phenformin (Phenformin).
  • a composition for use in the prevention or treatment of secondary cancer comprising an aldehyde inhibitor and a biguanide-based compound as an active ingredient, wherein the aldehyde inhibitor is Gosipol ( Gossypol) provides a composition for use in the prevention or treatment of secondary cancer, characterized in that the biguanide-based compound is phenformin (Phenformin) Provides a composition for use in the prevention or treatment of secondary cancer do.
  • Gosipol Gossypol
  • Phenformin phenformin
  • Secondary cancer refers to secondary cancer that has metastasized to a new organ by first metastasizing cancer from another site, and has a poorer prognosis than primary cancer.
  • the present invention relates to a pharmaceutical composition for the prevention and treatment of secondary cancer comprising an aldehyde inhibitor and a biguanide-based compound. It is expected to be widely used in the treatment of secondary cancer, which is difficult and has a poor prognosis.
  • FIGS. 2A and 2B are diagrams showing the cancer treatment effect of administration of gosipol and/or phenformin to primary or secondary colorectal cancer cell lines according to an embodiment of the present invention.
  • C denotes a control group
  • G denotes gossypol
  • P denotes phenformin
  • G+P denotes co-administration of gossypol and phenformin.
  • FIG. 3 is a diagram confirming the mitochondrial-dependent cell death effect of administration of gosipol and/or phenformin on primary or secondary colorectal cancer cell lines according to an embodiment of the present invention.
  • FIGS. 5A and 5B are diagrams showing the ATP synthesis inhibitory effect of administration of gosipol and/or phenformin to primary or secondary colorectal cancer cell lines according to an embodiment of the present invention.
  • C denotes a control
  • G denotes gossypol
  • P denotes phenformin
  • G+P denotes the combined administration of gossypol and phenformin.
  • FIGS. 6A and 6B are diagrams confirming the lactic acid generation results of administration of gossypol and/or phenformin to primary or secondary colorectal cancer cell lines according to an embodiment of the present invention.
  • C denotes gossypol as a control
  • P denotes phenformin
  • G+P denotes co-administration of gossypol and phenformin.
  • the present inventors promote cancer cell death when administered in combination with gosipol + phenformin in secondary colon cancer compared to primary colorectal cancer It was confirmed that the effect, the mitochondrial ATP synthesis inhibitory effect, and the lactic acid generation inhibitory effect were significant. From this, it was found that secondary colorectal cancer could be effectively treated by the combined administration of gosipol + phenformin.
  • the inventors of the present invention obtained primary colorectal cancer cell lines (SW480), and secondary colorectal cancer cell lines (LOVO, and SNU407) from Korea Cell Line Bank (KCLB), or American Type Culture Collection (ATCC). obtained and cultured according to the guidelines of KCLB or ATCC. Specific cell line information is shown in Table 1 below.
  • the results of anticancer treatment were confirmed by administering gosipol alone or in combination with phenformin to the colorectal cancer cell line of Example 1. More specifically, each cell was aliquoted in a 96-well plate at a concentration of 1 ⁇ 10 4 cell/well, and cultured overnight at 37° C., 5% CO 2 environment. Thereafter, gosypol was administered alone at a concentration of 0 to 16 ⁇ M, or co-administered with 10 ⁇ M or 100 ⁇ M of phenformin, and further cultured for 24 hours at 37° C., 5% CO 2 environment.
  • the cell viability results in the case of administering 1 ⁇ M of gosypol, 100 ⁇ M of phenformin, or a combination thereof are shown in FIG. 2 .
  • the primary colorectal cancer cell line, SW480 had no effect of co-administration (statistically insignificant), but the secondary colorectal cancer cell line, LOVO, had an effect of co-administration of gosipol and phenformin.
  • the cell viability test results were identical to the results of the cell viability experiment, and all of the groups administered with Gosipol alone, Gosipol + 10 ⁇ M phenformin, or Gosypol + 100 ⁇ M Fenformin had an ATP synthesis inhibitory effect, but in the LOVO cell line, SW480 The effect of drug administration was much more pronounced than that of the cell line.
  • Example 2 When gosipol 1 ⁇ M, phenformin 100 ⁇ M, or a combination thereof was administered, the primary colorectal cancer cell line, SW480, had no effect (statistically insignificant), but the secondary colorectal cancer cell line, LOVO, had no effect. The effect of concomitant administration of min was very pronounced. Therefore, it can be seen that the apoptosis effect in Example 2 is the effect of ATP synthesis inhibition in mitochondria.
  • Example 4 Cell cycle analysis of colorectal cancer cell lines treated with gossypol and/or phenformin
  • the ratio of cells for each cell cycle was analyzed. More specifically, each cell was washed twice with PBS, fixed with 70% ethanol/PBS, treated with RNase of 0.5 mg/ml dissolved in PBS containing 0.1% saponin, and reacted at 37° C. for 30 minutes , and stained with PI of 20 ⁇ g/ml at 4°C for 30 min.
  • the DNA content of cells was analyzed in 1 ⁇ 10 6 cells with a FACSCalibur flow cytometer and CellQuest software. The results are shown in Tables 2 to 4 below.
  • the primary colorectal cancer cell line (SW480), or the secondary colorectal cancer cell line (LOVO, and SNU407) was treated with 1 ⁇ M of gosipol and/or 100 ⁇ M of phenformin in the same manner as in Example 2, the lactic acid generation effect was confirmed did. More specifically, each cell was seeded in a 96 or 24-well plate at a concentration of 1 ⁇ 10 4 , or 3 ⁇ 10 5 cells/well, respectively, and cultured overnight, then the drug was administered and further cultured for 24 hours.
  • Secondary cancer refers to secondary cancer that has metastasized to a new organ by first metastasizing cancer from another site, and has a poorer prognosis than primary cancer.
  • the pharmaceutical composition of the present invention has a significant therapeutic effect in secondary cancer specifically compared to primary cancer, it is expected to be widely used in the treatment of secondary cancer, which is difficult to treat and has a poor prognosis.

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Abstract

The present invention relates to a pharmaceutical composition for prevention and treatment of a secondary cancer, comprising an aldehyde inhibitor and a biguanide-based compound. A secondary cancer refers to a cancer which results from the metastasis of a cancer starting in one place to a different organ, and exhibits more unfavorable prognosis than the primary cancer. The pharmaceutical composition according to the present invention comprises an aldehyde inhibitor and a biguanide-based compound for prevention and treatment of a secondary cancer. Having a more remarkable therapeutic effect specifically on a secondary cancer than a primary cancer, the pharmaceutical composition of the present invention is expected to find great applications in the treatment of a secondary cancer that is difficult to cure and unfavorable in prognosis.

Description

알데히드 억제제 및 비구아나이드 계열 화합물을 포함하는 속발성 암의 예방 및 치료용 약학조성물Pharmaceutical composition for the prevention and treatment of secondary cancer comprising an aldehyde inhibitor and a biguanide-based compound
본 발명은 알데히드 억제제 및 비구아나이드 계열 화합물을 포함하는 속발성 암의 예방 및 치료용 약학조성물에 관한 것이다.The present invention relates to a pharmaceutical composition for the prevention and treatment of secondary cancer comprising an aldehyde inhibitor and a biguanide-based compound.
대장암은 대장에 생기는 악성종양으로, 세계보건기구(WHO) 산하 국제암연구소(IARC) 보고서에 따르면, 한국의 대장암 발병률은 10만 명당 45명으로 세계 1위이다. 중앙암등록본부에서 발표한 암 등록 통계를 보면 2014년에 우리나라에서는 217,057건의 암이 발생했는데, 그 중 대장암은 26,978건으로 전체의 12.4%로 3위를 차지할 정도로 발병률이 높다. 대장암으로 인한 사망률 또한 암종별 사망자수의 4위를 차지할 정도로 높다. 이러한 대장암은 기대수명 증가와 서구화된 식습관에 의해 발병률의 증가가 더욱 가속화되고 있다. 그에 따라 많은 사회 비용이 초래되며 대장암 치료에 사용할 수 있는 약제의 수가 많지 않아 대장암에 대한 적극적인 연구가 반드시 필요하다. 대장암에는 현재 옥살리플라틴, 이리노테칸 같은 항암제와 세툭시맙이나 베바시주맙 같은 항체 치료제를 병용하여 사용하고 있으나, 대장암은 소화기계의 특성상 약물치료의 효과가 현저하지 않아, 대부분 수술로 이루어지고 있는 실정이다. 따라서 체력적으로 부담이 있는 노약자나 수술이 불가능한 환자는 치료가 여의치 않은 문제점이 있다. 또한 수술로 대장을 절제하면 암이 치료된 이후에도 소화기 능력이 저하되어 삶이 질이 떨어지는 문제점이 있으므로, 효과적인 암 치료제의 개발이 요구되는 실정이다.Colorectal cancer is a malignant tumor that occurs in the colon. According to a report by the International Agency for Research on Cancer (IARC) under the World Health Organization (WHO), the incidence rate of colorectal cancer in Korea is 45 per 100,000 people, the highest in the world. According to the cancer registration statistics announced by the Central Cancer Registry, there were 217,057 cancer cases in Korea in 2014, of which 26,978 cases were colorectal cancer, accounting for 12.4% of the total, taking the third place with a high incidence rate. Mortality due to colorectal cancer is also high enough to occupy the fourth place in the number of deaths by cancer type. The increase in the incidence of colorectal cancer is further accelerated by the increase in life expectancy and the westernized diet. As a result, many social costs are incurred, and there are not many drugs that can be used to treat colorectal cancer, so active research on colorectal cancer is absolutely necessary. Currently, anticancer drugs such as oxaliplatin and irinotecan are used in combination with antibody therapeutics such as cetuximab or bevacizumab. However, in colorectal cancer, the effect of drug treatment is not remarkable due to the characteristics of the digestive system. to be. Therefore, there is a problem in that it is difficult to treat the elderly who are physically burdened or patients who cannot operate. In addition, when the large intestine is surgically resected, the digestive capacity is lowered even after the cancer is treated, and there is a problem that the quality of life is deteriorated. Therefore, the development of an effective cancer treatment is required.
한편, 속발성 암은 다른 부위에서 처음 발생한 암이 전이되어 새로운 장기로 전이한 이차성 암을 의미하는 것으로서, 원발성 암에 비하여 예후가 불량하다.On the other hand, secondary cancer refers to a secondary cancer that has metastasized to a new organ by first metastasizing cancer from another site, and has a poorer prognosis than primary cancer.
본 발명은 상기와 같은 문제를 해결하기 위하여 안출된 것으로, 알데히드 억제제 및 비구아나이드 계열 화합물을 포함하는 속발성 암의 예방 및 치료용 약학조성물에 관한 것이다. 본 발명의 약학조성물은 원발성 암에 비하여 특이적으로 속발성 암에서 치료 효과가 현저하므로, 치료가 어렵고 예후가 불량한 속발성 암의 치료에 크게 이용될 것으로 기대된다.The present invention has been devised to solve the above problems, and relates to a pharmaceutical composition for the prevention and treatment of secondary cancer comprising an aldehyde inhibitor and a biguanide-based compound. Since the pharmaceutical composition of the present invention has a significant therapeutic effect in secondary cancer specifically compared to primary cancer, it is expected to be widely used in the treatment of secondary cancer, which is difficult to treat and has a poor prognosis.
본 발명은 알데히드 억제제 및 비구아나이드 계열 화합물을 포함하는 속발성 암의 예방 및 치료용 약학조성물에 관한 것이다.The present invention relates to a pharmaceutical composition for the prevention and treatment of secondary cancer comprising an aldehyde inhibitor and a biguanide-based compound.
그러나 본 발명이 이루고자 하는 기술적 과제는 이상에서 언급한 과제에 제한되지 않으며, 언급되지 않은 또 다른 과제들은 아래의 기재로부터 당 업계에서 통상의 지식을 가진 자에게 명확하게 이해될 수 있을 것이다.However, the technical task to be achieved by the present invention is not limited to the tasks mentioned above, and other tasks not mentioned will be clearly understood by those of ordinary skill in the art from the following description.
이하, 본원에 기재된 다양한 구체예가 도면을 참조로 기재된다. 하기 설명에서, 본 발명의 완전한 이해를 위해서, 다양한 특이적 상세사항, 예컨대, 특이적 형태, 조성물 및 공정 등이 기재되어 있다. 그러나, 특정의 구체예는 이들 특이적 상세 사항 중 하나 이상 없이, 또는 다른 공지된 방법 및 형태와 함께 실행될 수 있다. 다른 예에서, 공지된 공정 및 제조 기술은 본 발명을 불필요하게 모호하게 하지 않게 하기 위해서, 특정의 상세사항으로 기재되지 않는다. "한 가지 구체예" 또는 "구체예"에 대한 본 명세서 전체를 통한 참조는 구체예와 결부되어 기재된 특별한 특징, 형태, 조성 또는 특성이 본 발명의 하나 이상의 구체예에 포함됨을 의미한다. 따라서, 본 명세서 전체에 걸친 다양한 위치에서 표현된 "한 가지 구체예에서" 또는 "구체예"의 상황은 반드시 본 발명의 동일한 구체예를 나타내지는 않는다. 추가로, 특별한 특징, 형태, 조성, 또는 특성은 하나 이상의 구체예에서 어떠한 적합한 방법으로 조합될 수 있다.Hereinafter, various embodiments described herein are described with reference to the drawings. In the following description, various specific details are set forth, such as specific forms, compositions and processes, and the like, for a thorough understanding of the present invention. However, certain embodiments may be practiced without one or more of these specific details, or in conjunction with other known methods and forms. In other instances, well-known processes and manufacturing techniques have not been described in specific detail in order not to unnecessarily obscure the present invention. Reference throughout this specification to “one embodiment” or “an embodiment” means that a particular feature, form, composition, or characteristic described in connection with the embodiment is included in one or more embodiments of the invention. Thus, references to "in one embodiment" or "an embodiment" in various places throughout this specification do not necessarily refer to the same embodiment of the invention. Additionally, the particular features, forms, compositions, or properties may be combined in any suitable manner in one or more embodiments.
명세서에서 특별한 정의가 없으면 본 명세서에 사용된 모든 과학적 및 기술적인 용어는 본 발명이 속하는 기술분야에서 당업자에 의하여 통상적으로 이해되는 것과 동일한 의미를 가진다.Unless otherwise defined in the specification, all scientific and technical terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
본 발명의 일 구체예에서 “암”이란, 제어되지 않은 세포성장으로 특징지어지며, 이러한 비정상적인 세포성장에 의해 종양이라고 불리는 세포 덩어리가 형성되어 주위의 조직으로 침투하고 심한 경우에는 신체의 다른 기관으로 전이되기도 하는 것을 말한다. 학문적으로는 신생물이라고 명명되기도 한다. 암은 수술, 방사선 및 화학요법으로 치료를 하더라도 많은 경우에 근본적인 치유가 되지 못하고 환자에게 고통을 주며 궁극적으로는 죽음에 이르게 하는 난치성 만성질환으로, 암의 발생요인으로는 여러 가지가 있으나, 내적 요인과 외적 요인으로 구분한다. 정상세포가 어떠한 기전을 거처 암세포로 형질전환이 되는지에 대해서는 정확하게 규명되지 않았으나, 상당수의 암이 환경요인 등 외적인자에 의해 영향을 받아 발생하는 것으로 알려져 있다. 내적 요인으로는 유전 인자, 면역학적 요인 등이 있으며, 외적 요인으로는 화학물질, 방사선, 바이러스 등이 있다. 암의 발생에 관련되는 유전자에는 종양형성유전자 (oncogenes)와 종양억제유전자 (tumor suppressor genes)가 있는데, 이들 사이의 균형이 위에서 설명한 내적 혹은 외적 용인들에 의해 무너질 때 암이 발생하게 된다.In one embodiment of the present invention, "cancer" is characterized by uncontrolled cell growth, and by this abnormal cell growth, a cell mass called a tumor is formed, penetrates into surrounding tissues, and in severe cases, it spreads to other organs of the body. It is said to be transferable. Scientifically, it is also called a neoplasm. Cancer is an intractable chronic disease that in many cases cannot be cured fundamentally even if treated with surgery, radiation, and chemotherapy, but causes pain and ultimately death. There are several causes of cancer, but internal factors and external factors. It is not known exactly how normal cells are transformed into cancer cells, but it is known that a significant number of cancers are affected by external factors such as environmental factors. Internal factors include genetic factors and immunological factors, and external factors include chemicals, radiation, and viruses. The genes involved in the development of cancer include oncogenes and tumor suppressor genes, and cancer occurs when the balance between them is disrupted by the internal or external factors described above.
본 발명의 일 구체예에서 “원발성 암(Primary cancer)”란 원래 그 자리에서 발생한 암을 의미한다. 해당 장기에서 최초로 발생하였다는 의미를 함께 내포한다. 상대적 용어로 속발성 암(Secondary cancer)과 구분한다. 예를 들어, 대장에 원래 있던 세포가 암세포로 변한 “원발성 대장암”을 들 수 있다.In one embodiment of the present invention, “primary cancer” refers to a cancer that originally occurred in situ. It also implies that it occurred first in the organ. In relative terms, it is distinguished from secondary cancer. For example, there is “primary colorectal cancer” in which cells that were originally in the colon have turned into cancer cells.
본 발명의 일 구체예에서 “속발성 암(Secondary cancer)”이란, 상기 원발성 암과 구분되는 의므로서, 다른 원인으로 생긴 결과로서의 암을 의미한다. 예를 들어, 다른 곳에서 생긴 암세포가 대장에 전이되어 정착한 뒤 암 조직이 자라는 경우에는 “속발성 대장암”이라고 한다.In one embodiment of the present invention, “secondary cancer” is a meaning distinct from the primary cancer, and refers to cancer as a result of other causes. For example, if cancer cells originating elsewhere metastasize to the colon and colonize and then grow cancerous tissue, it is called “secondary colorectal cancer”.
본 발명의 일 구체예에서 “암 전이(cancer metastasis)”란, 암세포가 원발 장기를 떠나 다른 장기로 가는 것이며, 상기의 “암”이란 “암 줄기세포”를 포함하는 의미이다. 암이 신체의 다른 부분으로 퍼지는 것은 크게 원발암에서 암조직이 성장하여 직접적으로 주위 장기를 침윤하는 것과, 멀리 있는 다른 장기로 혈관이나 림프관을 따라 원격 전이를 하는 것으로 나눈다.In one embodiment of the present invention, “cancer metastasis” means that cancer cells leave a primary organ and go to another organ, and the term “cancer” includes “cancer stem cells”. The spread of cancer to other parts of the body is largely divided into growth of cancer tissue from the primary cancer and directly infiltrating surrounding organs, and distant metastasis along blood vessels or lymphatic vessels to other distant organs.
본 발명의 일 구체예에서 “치료”란, 목적하는 질병의 완화 또는/및 개선을 위해 수행되는 일련의 활동을 의미한다. 본 발명의 목적상 치료는 속발성 암에 있어서 암 줄기세포를 포함하는 암세포의 수적 또는 양적 증식을 억제시키거나, 암세포를 사멸시키거나, 암 조직의 성장을 억제시키거나, 암 조직의 크기를 감소시키거나, 암 조직 내의 신생혈관의 발달을 억제시키거나, 암이 또 다른 부위로 침윤 또는 전이되는 것을 억제하는 활동을 포함한다.In one embodiment of the present invention, “treatment” refers to a series of activities performed for alleviation and/or amelioration of a desired disease. For the purpose of the present invention, treatment is to inhibit numerical or quantitative proliferation of cancer cells including cancer stem cells in secondary cancer, kill cancer cells, inhibit the growth of cancer tissues, or reduce the size of cancer tissues. or inhibiting the development of new blood vessels in cancer tissue, or inhibiting the invasion or metastasis of cancer to another site.
본 발명자들은 연구 결과, 알데히드 억제제(aldehyde inhibitor) 및 비구아나이드(biguanide) 계열 화합물을 병용 투여할 경우, 속발성 암에 특이적으로 암 치료 효과가 현저함을 확인하여 본 발명을 완성하게 되었다.The present inventors have completed the present invention by confirming that, as a result of the study, when an aldehyde inhibitor and a biguanide-based compound are administered in combination, the cancer treatment effect specifically for secondary cancer is remarkable.
구체적으로, 본 발명은 알데히드 억제제(aldehyde inhibitor) 및 비구아나이드(biguanide) 계열 화합물을 포함하는 속발성 암의 예방 또는 치료용 약학조성물에 관한 것으로, 바람직하게는 상기 알데히드 억제제는 3-히드록시키누레닌(3-hydroxy-DL-kynurenine; 3-HDK), 베노밀(benomyl), 시스플라틴(cis-diamminedichloridoplatinum; CDDP), 클로르프로파마이드(chlorpropamide), 시트랄(citral), CVT-10216(3-[[[3-[4-[(methylsulfonyl)amino]phenyl]-4-oxo-4H-1-benzopyran-7-yl]oxy]methyl]-benzoic acid, 또는 3-[[[3-[4-[(methylsulfonyl)amino]phenyl]-4-oxo-4H-chromen-7-yl]oxy]methyl]benzoic acid), 사이안아마이드(cyanamide), 다이진(daidzin), 디에틸아미노벤즈알데히드(Diethylaminobenzaldehyde; DEAB), 디설피람(disulfiram), 고시폴(gossypol), 키누렌산(kynurenic acid), 모리네이트(molinate), 파르질린(pargyline), 포스포(에놀)피루브산 모노나트륨염 수화물(phospho(enol)pyruvic acid monosodium salt hydrate), 페닐글리옥산(phenylglyoxal), 레티노산(retinoic acid), N-아세틸-N-아세톡시-4-클로로벤젠설폰아미드(N-acetyl-N-acetoxy-4-chlorobenzenesulfonamide), 및 옥살산나트륨(sodium oxamate)으로 구성되는 그룹으로부터 선택되는 어느 하나일 수 있고, 보다 바람직하게는 고시폴(gossypol)일 수 있다. 상기 비구아나이드 계열 화합물은 바람직하게는 메트포민(metformin), 펜포르민(phenformin), 또는 부포르민(buformine)일 수 있으나, 보다 바람직하게는 펜포르민(phenformin)일 수 있다.Specifically, the present invention relates to a pharmaceutical composition for preventing or treating secondary cancer comprising an aldehyde inhibitor and a biguanide-based compound, preferably the aldehyde inhibitor is 3-hydroxykynurenine ( 3-hydroxy-DL-kynurenine; 3-HDK), benomyl, cis-diamminedichloridoplatinum (CDDP), chlorpropamide, citral, CVT-10216 (3-[[[ 3-[4-[(methylsulfonyl)amino]phenyl]-4-oxo-4H-1-benzopyran-7-yl]oxy]methyl]-benzoic acid, or 3-[[[3-[4-[(methylsulfonyl )amino]phenyl]-4-oxo-4H-chromen-7-yl]oxy]methyl]benzoic acid), cyanamide, daidzin, diethylaminobenzaldehyde (DEAB), diethylaminobenzaldehyde disulfiram, gossypol, kynurenic acid, molinate, pargyline, phospho(enol)pyruvic acid monosodium salt hydrate), phenylglyoxal, retinoic acid, N-acetyl-N-acetoxy-4-chlorobenzenesulfonamide, and sodium oxalate ( sodium oxamate) may be any one selected from the group consisting of, more preferably gossypol (gossypol). The biguanide-based compound may be preferably metformin, phenformin, or buformine, but more preferably phenformin.
또한, 본 발명의 약학조성물에서 상기 알데히드 억제제와 비구아나이드 계열 화합물은 1:1~100의 중량비로 포함될 수 있고, 바람직하게는 1:2~20의 중량비로 포함될 수 있다.In addition, in the pharmaceutical composition of the present invention, the aldehyde inhibitor and the biguanide-based compound may be included in a weight ratio of 1:1 to 100, preferably in a weight ratio of 1:2 to 20.
또한, 본 발명의 약학조성물에서 상기 알데히드 억제제는 0.5~50μM의 양으로 포함될 수 있다.In addition, in the pharmaceutical composition of the present invention, the aldehyde inhibitor may be included in an amount of 0.5-50 μM.
또한, 본 발명의 약학조성물에서 상기 비구아나이드 계열 화합물은 10~1000μM의 양으로 포함될 수 있다.In addition, in the pharmaceutical composition of the present invention, the biguanide-based compound may be included in an amount of 10 to 1000 μM.
상기한 바와 같이, 본 발명의 조성물은 속발성 암을 억제하여 이를 예방 및/또는 치료할 수 있고, 바람직하게는 속발성 대장암을 효과적으로 예방 및/또는 치료할 수 있는 것을 특징으로 한다.As described above, the composition of the present invention can prevent and/or treat secondary cancer by inhibiting it, preferably, it is characterized in that it can effectively prevent and/or treat secondary colorectal cancer.
다만, 본 발명의 약학조성물은 추가로 다른 항암제와 병용 투여할 수 있으며, 이를 통해서 암 줄기세포뿐만 아니라 일반적인 암세포까지 효과적으로 치료하는데 사용될 수 있다. However, the pharmaceutical composition of the present invention may be additionally administered in combination with other anticancer agents, and through this, it can be used to effectively treat cancer stem cells as well as general cancer cells.
여기서 상기 항암제로는 나이트로젠 머스타드, 이마티닙, 옥살리플라틴, 리툭시맙, 엘로티닙, 네라티닙, 라파티닙, 제피티닙, 반데타닙, 니로티닙, 세마사닙, 보수티닙, 악시티닙, 세디라닙, 레스타우르티닙, 트라스투주맙, 게피티니브, 보르테조밉, 수니티닙, 카보플라틴, 소라페닙, 베바시주맙, 시스플라틴, 세툭시맙, 비스쿰알붐, 아스파라기나제, 트레티노인, 하이드록시카바마이드, 다사티닙, 에스트라머스틴, 겜투주맵오조가마이신, 이브리투모맙튜세탄, 헵타플라틴, 메칠아미노레불린산, 암사크린, 알렘투주맙, 프로카르바진, 알프로스타딜, 질산홀뮴 키토산, 젬시타빈, 독시플루리딘, 페메트렉세드, 테가푸르, 카페시타빈, 기메라신, 오테라실, 아자시티딘, 메토트렉세이트, 우라실, 시타라빈, 플루오로우라실, 플루다가빈, 에노시타빈, 플루타미드, 데시타빈, 머캅토푸린, 티오구아닌, 클라드리빈, 카르모퍼, 랄티트렉세드, 도세탁셀, 파클리탁셀, 이리노테칸, 벨로테칸, 토포테칸, 비노렐빈, 에토포시드, 빈크리스틴, 빈블라스틴, 테니포시드, 독소루비신, 이다루비신, 에피루비신, 미톡산트론, 미토마이신, 블레로마이신, 다우노루비신, 닥티노마이신, 피라루비신, 아클라루비신, 페프로마이신, 템시롤리무스, 테모졸로마이드, 부설판, 이포스파미드, 사이클로포스파미드, 멜파란, 알트레트민, 다카바진, 치오테파, 니무스틴, 클로람부실, 미토락톨, 레우코보린, 트레토닌, 엑스메스탄, 아미노글루테시미드, 아나그렐리드, 나벨빈, 파드라졸, 타목시펜, 토레미펜, 테스토락톤, 아나스트로졸, 레트로졸, 보로졸, 비칼루타미드, 로무스틴 및 카르무스틴으로 이루어진 군에서 선택된 1종 이상을 사용할 수 있으나, 이에 한정되는 것은 아니다.Here, the anticancer agent is nitrogen mustard, imatinib, oxaliplatin, rituximab, erlotinib, neratinib, lapatinib, gefitinib, vandetanib, nirotinib, semasanib, bosutinib, axitinib, cediranib , restautinib, trastuzumab, gefitinib, bortezomib, sunitinib, carboplatin, sorafenib, bevacizumab, cisplatin, cetuximab, viscumalbum, asparaginase, tretinoin, hydroxycarba amide, dasatinib, estramustine, gemtuzumab ozogamicin, ibritumomab tuccetan, heptaplatin, methylaminolevulinic acid, amsacrine, alemtuzumab, procarbazine, alprostadil, holmium nitrate Chitosan, gemcitabine, doxyfluridine, pemetrexed, tegafur, capecitabine, gimeracin, oteracil, azacitidine, methotrexate, uracil, cytarabine, fluorouracil, fludagabine, enocy Tabine, flutamide, decitabine, mercaptopurine, thioguanine, cladribine, carmopher, raltitrexed, docetaxel, paclitaxel, irinotecan, belotecan, topotecan, vinorelbine, etoposide, vincristine, vin Blastin, teniposide, doxorubicin, idarubicin, epirubicin, mitoxantrone, mitomycin, bleromycin, daunorubicin, dactinomycin, pyrarubicin, aclarubicin, pepromycin, temsi Rolimus, temozolomide, busulfan, ifosfamide, cyclophosphamide, melparan, altretmine, dacarbazine, thiotepa, nimustine, chlorambucil, mitolactol, leucovorin, tretonin, X as mestan, aminoglutethimide, anagrelide, nabelbine, fadrazole, tamoxifen, toremifene, testolactone, anastrozole, letrozole, vorozole, bicalutamide, lomustine and carmustine One or more selected from the group consisting of may be used, but is not limited thereto.
본 발명에 있어서, 상기 약학조성물은 캡슐, 정제, 과립, 주사제, 연고제, 분말 또는 음료 형태임을 특징으로 할 수 있으며, 상기 약학조성물은 인간을 대상으로 하는 것을 특징으로 할 수 있다. In the present invention, the pharmaceutical composition may be characterized in the form of capsules, tablets, granules, injections, ointments, powders or beverages, and the pharmaceutical composition may be characterized in that it is intended for humans.
본 발명의 약학조성물은 이들로 한정되는 것은 아니지만, 각각 통상의 방법에 따라 산제, 과립제, 캡슐, 정제, 수성 현탁액 등의 경구형 제형, 외용제, 좌제 및 멸균 주사용액의 형태로 제형화하여 사용될 수 있다. 본 발명의 약학조성물은 약제적으로 허용 가능한 담체를 포함할 수 있다. 약제학적으로 허용되는 담체는 경구 투여 시에는 결합제, 활탁제, 붕해제, 부형제, 가용화제, 분산제, 안정화제, 현탁화제, 색소, 향료 등을 사용할 수 있으며, 주사제의 경우에는 완충제, 보존제, 무통화제, 가용화제, 등장제, 안정화제 등을 혼합하여 사용할 수 있으며, 국소투여용의 경우에는 기제, 부형제, 윤활제, 보존제 등을 사용할 수 있다. 본 발명의 약제학적 조성물의 제형은 상술한 바와 같은 약제학적으로 허용되는 담체와 혼합하여 다양하게 제조될 수 있다. 예를 들어, 경구 투여시에는 정제, 트로키, 캡슐, 엘릭서(elixir), 서스펜션, 시럽, 웨이퍼 등의 형태로 제조할 수 있으며, 주사제의 경우에는 단위 투약 앰플 또는 다수회 투약 형태로 제조할 수 있다. 기타, 용액, 현탁액, 정제, 캡슐, 서방형 제제 등으로 제형할 수 있다.The pharmaceutical composition of the present invention is not limited thereto, but can be formulated and used in the form of oral dosage forms such as powders, granules, capsules, tablets, aqueous suspensions, external preparations, suppositories, and sterile injection solutions, respectively, according to conventional methods. have. The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers may include binders, lubricants, disintegrants, excipients, solubilizers, dispersants, stabilizers, suspending agents, dyes, fragrances, etc., for oral administration, and in the case of injections, buffers, preservatives, pain-freezing agents A topical agent, solubilizer, isotonic agent, stabilizer, etc. may be mixed and used, and in the case of topical administration, a base, excipient, lubricant, preservative, etc. may be used. The dosage form of the pharmaceutical composition of the present invention can be prepared in various ways by mixing with a pharmaceutically acceptable carrier as described above. For example, in the case of oral administration, it can be prepared in the form of tablets, troches, capsules, elixirs, suspensions, syrups, wafers, etc., and in the case of injections, it can be prepared in the form of unit dose ampoules or multiple doses. have. In addition, it can be formulated as a solution, suspension, tablet, capsule, sustained release formulation, and the like.
한편, 제제화에 적합한 담체, 부형제 및 희석제의 예로는, 락토즈, 덱스트로즈, 수크로즈, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말디톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로즈, 메틸 셀룰로즈, 미정질 셀룰로즈, 폴리비닐피롤리돈, 물, 메틸하이드록시벤조에이트, 프로필하이드록시벤조에이트, 탈크, 마그네슘 스테아레이트 또는 광물유 등이 사용될 수 있다. 또한, 충진제, 항응집제, 윤활제, 습윤제, 향료, 유화제, 방부제 등을 추가로 포함할 수 있다.Meanwhile, examples of suitable carriers, excipients and diluents for formulation include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, malditol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate or mineral oil may be used. In addition, fillers, anti-agglomeration agents, lubricants, wetting agents, fragrances, emulsifiers, preservatives and the like may be further included.
본 발명에 따른 약학조성물의 투여 경로는 이들로 한정되는 것은 아니지만 구강, 정맥내, 근육내, 동맥내, 골수내, 경막내, 심장내, 경피, 피하, 복강내, 비강내, 장관, 국소, 설하 또는 직장이 포함된다. 경구 또는 비경구 투하가 바람직하다. 본원에 사용된 용어 "비경구"는 피하, 피내, 정맥내, 근육내, 관절내, 활액낭내, 흉골내, 경막내, 병소내 및 두개골내 주사 또는 주입기술을 포함한다. 본 발명의 약학조성물은 또한 직장 투여를 위한 좌제의 형태로 투여될 수 있다.The route of administration of the pharmaceutical composition according to the present invention is, but not limited to, oral, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, intestinal, topical, sublingual or rectal. Oral or parenteral administration is preferred. As used herein, the term “parenteral” includes subcutaneous, intradermal, intravenous, intramuscular, intraarticular, intrabursar, intrasternal, intrathecal, intralesional and intracranial injection or infusion techniques. The pharmaceutical composition of the present invention may also be administered in the form of a suppository for rectal administration.
본 발명의 약학조성물은 사용된 특정 화합물의 활성, 연령, 체중, 일반적인 건강, 성별, 정식, 투여시간, 투여경로, 배출율, 약물 배합 및 예방 또는 치료될 특정 질환의 중증을 포함한 여러 요인에 따라 다양하게 변할 수 있고, 상기 약학조성물의 투여량은 환자의 상태, 체중, 질병의 정도, 약무형태, 투여경로 및 기간에 따라 다르지만 당업자에 의해 적절하게 선택될 수 있고, 1일 0.0001 내지 50mg/kg 또는 0.001 내지 50mg/kg으로 투여할 수 있다. 투여는 하루에 한번 투여할 수도 있고, 수회 나누어 투여할 수도 있다. 상기 투여량은 어떠한 면으로든 본 발명의 범위를 한정하는 것은 아니다. 본 발명에 따른 의약 조성물은 환제, 당의정, 캡슐, 액제, 겔, 시럽, 슬러리, 현탁제로 제형될 수 있다.The pharmaceutical composition of the present invention varies depending on several factors including the activity of the specific compound used, age, weight, general health, sex, formula, administration time, administration route, excretion rate, drug formulation, and the severity of the specific disease to be prevented or treated The dosage of the pharmaceutical composition may vary depending on the patient's condition, weight, degree of disease, drug form, administration route and period, but may be appropriately selected by those skilled in the art, and 0.0001 to 50 mg/kg per day or It may be administered at 0.001 to 50 mg/kg. Administration may be administered once a day, or may be administered in several divided doses. The above dosage does not limit the scope of the present invention in any way. The pharmaceutical composition according to the present invention may be formulated as pills, dragees, capsules, solutions, gels, syrups, slurries, and suspensions.
본 발명의 일 구체예에서 “식품 조성물”이란, 본 발명에서 목적으로 하는 적응증의 예방 또는 개선을 위해 다양하게 이용되는 것으로서, 본 발명의 조성물을 유효성분으로 포함하는 식품조성물은 각종 식품류, 예를 들어, 음료, 껌, 차, 비타민 복합제, 분말, 과립, 정제, 캡슐, 과자, 떡, 빵 등의 형태로 제조될 수 있다. 본 발명의 식품조성물은 본 발명의 적응증을 예방하기 위한 목적으로 장기간 복용 시에도 안심하고 사용할 수 있다. 본 발명의 조성물이 식품조성물에 포함될 때 그 양은 전체 중량의 0.1 내지 100%의 비율로 첨가할 수 있다. 여기서, 상기 식품조성물이 음료 형태로 제조되는 경우 지시된 비율로 상기 식품조성물을 함유하는 것 외에 특별한 제한점은 없으며 통상의 음료와 같이 여러가지 향미제 또는 천연탄수화물 등을 추가 성분으로서 함유할 수 있다. 즉, 천연탄수화물로서 포도당 등의 모노사카라이드, 과당 등의 디사카라이드, 슈크로스 등의 및 폴리사카라이드, 덱스트린, 시클로덱스트린 등과 같은 통상적인 당 및 자일리톨, 소르비톨, 에리트리톨 등의 당알콜 등을 포함할 수 있다. 상기 향미제로서는 천연 향미제(타우마틴, 스테비아 추출물(예를 들어 레바우디오시드 A, 글리시르히진등) 및 합성 향미제(사카린, 아스파르탐 등) 등을 들 수 있다. 그 외 본 발명의 식품조성물은 여러 가지 영양제, 비타민, 광물(전해질), 합성풍미제 및 천연풍미제 등의 풍미제, 착색제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산음료에 사용되는 탄산화제 등을 함유할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다. 이러한 첨가제의 비율은 통상적으로 본 발명의 조성물 100 중량부 당 0.1 내지 100 중량부의 범위에서 선택되는 것이 일반적이나, 이에 제한되는 것은 아니다.In one embodiment of the present invention, "food composition" is used in various ways for the prevention or improvement of indications for the purpose of the present invention, and the food composition comprising the composition of the present invention as an active ingredient includes various foods, such as For example, it may be prepared in the form of beverage, gum, tea, vitamin complex, powder, granule, tablet, capsule, confectionery, rice cake, bread, and the like. The food composition of the present invention can be safely used even when taken for a long time for the purpose of preventing the indications of the present invention. When the composition of the present invention is included in the food composition, the amount may be added in a proportion of 0.1 to 100% of the total weight. Here, when the food composition is prepared in the form of a beverage, there is no particular limitation other than containing the food composition in the indicated ratio, and it may contain various flavoring agents or natural carbohydrates as additional ingredients, like a conventional beverage. That is, as natural carbohydrates, monosaccharides such as glucose, disaccharides such as fructose, and polysaccharides such as sucrose, common sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol are included. can do. Examples of the flavoring agent include natural flavoring agents (taumatin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.). Others of the present invention of various nutrients, vitamins, minerals (electrolytes), synthetic and natural flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, It may contain stabilizer, preservative, glycerin, alcohol, carbonation agent used in carbonated beverage, etc. These components can be used independently or in combination.The proportion of these additives is usually per 100 parts by weight of the composition of the present invention. It is generally selected in the range of 0.1 to 100 parts by weight, but is not limited thereto.
본 발명의 일 구체예에서, 알데히드 억제제(aldehyde inhibitor) 및 비구아나이드(biguanide) 계열 화합물을 포함하는 속발성 암의 예방 또는 치료용 약학조성물을 제공하고, 상기 알데히드 억제제는 고시폴(Gossypol)인 것을 특징으로 하는 약학조성물을 제공하며, 상기 비구아나이드 계열 화합물은 펜포르민(Phenformin)인 것을 특징으로 하는 약학조성물을 제공하며, 상기 알데히드 억제제와 비구아나이드 계열 화합물은 1:1~100의 중량비로 포함되는 것을 특징으로 하는 약학조성물을 제공하며, 상기 알데히드 억제제와 비구아나이드 계열 화합물은 1:2~20의 중량비로 포함되는 것을 특징으로 하는 약학조성물을 제공하며, 상기 알데히드 억제제는 0.5 내지 50 μM의 양으로 포함되는 것을 특징으로 하는 약학조성물을 제공하며, 상기 비구아나이드 계열 화합물은 10 내지 1000 μM의 양으로 포함되는 것을 특징으로 하는 약학조성물을 제공하며, 상기 속발성 암은 속발성 대장암인 것을 특징으로 하는 약학조성물을 제공하며, 상기 속발성 암은 주변 장기로부터 침윤되어나, 또는 혈관 또는 림프절로부터 원격 전이된 것을 특징으로 하는 약학조성물을 제공하며, 상기 암은 암 줄기세포를 포함하는 것을 특징으로 하는 약학조성물을 제공한다.In one embodiment of the present invention, there is provided a pharmaceutical composition for preventing or treating secondary cancer comprising an aldehyde inhibitor and a biguanide-based compound, wherein the aldehyde inhibitor is Gossypol It provides a pharmaceutical composition comprising the above, wherein the biguanide-based compound is phenformin, wherein the aldehyde inhibitor and the biguanide-based compound are included in a weight ratio of 1:1 to 100. It provides a pharmaceutical composition, characterized in that, the aldehyde inhibitor and the biguanide-based compound provides a pharmaceutical composition characterized in that it is included in a weight ratio of 1: 2 to 20, the aldehyde inhibitor is in an amount of 0.5 to 50 μM It provides a pharmaceutical composition characterized in that it is included, the biguanide-based compound provides a pharmaceutical composition characterized in that it is included in an amount of 10 to 1000 μM, wherein the secondary cancer is secondary colorectal cancer. It provides a composition, wherein the secondary cancer has infiltrated from nearby organs or has spread remotely from blood vessels or lymph nodes, wherein the cancer comprises cancer stem cells. to provide.
본 발명의 또 다른 구체예에서, 알데히드 억제제(aldehyde inhibitor) 및 비구아나이드(biguanide) 계열 화합물을 유효성분으로 포함하는 약학조성물로 속발성 암을 예방 또는 치료하는 방법을 제공하고, 상기 알데히드 억제제는 고시폴(Gossypol)인 것을 특징으로 하는 속발성 암을 예방 또는 치료하는 방법을 제공하며, 상기 비구아나이드 계열 화합물은 펜포르민(Phenformin)인 것을 특징으로 하는 속발성 암을 예방 또는 치료하는 방법을 제공한다.In another embodiment of the present invention, there is provided a method for preventing or treating secondary cancer with a pharmaceutical composition comprising an aldehyde inhibitor and a biguanide-based compound as an active ingredient, wherein the aldehyde inhibitor is Gossypol (Gossypol) provides a method for preventing or treating secondary cancer, wherein the biguanide-based compound is phenformin (Phenformin).
본 발명의 또 다른 구체예에서, 알데히드 억제제(aldehyde inhibitor) 및 비구아나이드(biguanide) 계열 화합물의 속발성 암 예방 또는 치료 용도를 제공하고, 상기 알데히드 억제제는 고시폴(Gossypol)인 것을 특징으로 하는 속발성 암 예방 또는 치료 용도를 제공하며, 상기 비구아나이드 계열 화합물은 펜포르민(Phenformin)인 것을 특징으로 하는 속발성 암 예방 또는 치료 용도를 제공한다.In another embodiment of the present invention, there is provided a secondary cancer prevention or treatment use of an aldehyde inhibitor and a biguanide-based compound, wherein the aldehyde inhibitor is Gossypol secondary cancer It provides a preventive or therapeutic use, and the biguanide-based compound provides a secondary cancer prevention or treatment use, characterized in that the biguanide-based compound is phenformin.
본 발명의 또 다른 구체예에서, 알데히드 억제제(aldehyde inhibitor) 및 비구아나이드(biguanide) 계열 화합물을 유효성분으로 포함하는 약학조성물의 속발성 암 예방 또는 치료 용도를 제공하고, 상기 알데히드 억제제는 고시폴(Gossypol)인 것을 특징으로 하는 속발성 암 예방 또는 치료 용도를 제공하며, 상기 비구아나이드 계열 화합물은 펜포르민(Phenformin)인 것을 특징으로 하는 속발성 암 예방 또는 치료 용도를 제공한다.In another embodiment of the present invention, there is provided a secondary cancer prevention or treatment use of a pharmaceutical composition comprising an aldehyde inhibitor and a biguanide-based compound as an active ingredient, wherein the aldehyde inhibitor is Gossypol ) provides a secondary cancer prevention or treatment use, characterized in that the biguanide-based compound provides a secondary cancer prevention or treatment use, characterized in that the phenformin (Phenformin).
본 발명의 또 다른 구체예에서, 알데히드 억제제(aldehyde inhibitor) 및 비구아나이드(biguanide) 계열 화합물을 유효성분으로 포함하는 속발성 암 예방 또는 치료에 사용하기 위한 조성물을 제공하며, 상기 알데히드 억제제는 고시폴(Gossypol)인 것을 특징으로 하는 속발성 암 예방 또는 치료에 사용하기 위한 조성물을 제공하며, 상기 비구아나이드 계열 화합물은 펜포르민(Phenformin)인 것을 특징으로 하는 속발성 암 예방 또는 치료에 사용하기 위한 조성물을 제공한다.In another embodiment of the present invention, there is provided a composition for use in the prevention or treatment of secondary cancer comprising an aldehyde inhibitor and a biguanide-based compound as an active ingredient, wherein the aldehyde inhibitor is Gosipol ( Gossypol) provides a composition for use in the prevention or treatment of secondary cancer, characterized in that the biguanide-based compound is phenformin (Phenformin) Provides a composition for use in the prevention or treatment of secondary cancer do.
이하 상기 본 발명을 단계별로 상세히 설명한다.Hereinafter, the present invention will be described in detail step by step.
속발성 암은 다른 부위에서 처음 발생한 암이 전이되어 새로운 장기로 전이한 이차성 암을 의미하는 것으로서, 원발성 암에 비하여 예후가 불량하다.Secondary cancer refers to secondary cancer that has metastasized to a new organ by first metastasizing cancer from another site, and has a poorer prognosis than primary cancer.
본 발명은 알데히드 억제제 및 비구아나이드 계열 화합물을 포함하는 속발성 암의 예방 및 치료용 약학조성물에 관한 것으로, 본 발명의 약학조성물은 원발성 암에 비하여 특이적으로 속발성 암에서 치료 효과가 현저하므로, 치료가 어렵고 예후가 불량한 속발성 암의 치료에 크게 이용될 것으로 기대된다.The present invention relates to a pharmaceutical composition for the prevention and treatment of secondary cancer comprising an aldehyde inhibitor and a biguanide-based compound. It is expected to be widely used in the treatment of secondary cancer, which is difficult and has a poor prognosis.
도 1a, 도 1b, 도 2a, 및 도 2b는 본 발명의 일 실시예에 따른, 원발성 또는 속발성 대장암 세포주에 대한 고시폴 및/또는 펜포르민 투여의 암 치료 효과를 나타낸 도이다. 도 2a 및 도 2b에서 C는 대조군(control), G는 고시폴(gossypol), P는 펜포르민(phenformin), G+P는 고시폴과 펜포르민의 병용 투여를 나타낸다.1A, 1B, 2A, and 2B are diagrams showing the cancer treatment effect of administration of gosipol and/or phenformin to primary or secondary colorectal cancer cell lines according to an embodiment of the present invention. In FIGS. 2A and 2B , C denotes a control group, G denotes gossypol, P denotes phenformin, and G+P denotes co-administration of gossypol and phenformin.
도 3은 본 발명의 일 실시예에 따른, 원발성 또는 속발성 대장암 세포주에 대한 고시폴 및/또는 펜포르민 투여의 미토콘드리아 의존성 세포사 효과 확인한 도이다.3 is a diagram confirming the mitochondrial-dependent cell death effect of administration of gosipol and/or phenformin on primary or secondary colorectal cancer cell lines according to an embodiment of the present invention.
도 4a, 도 4b, 도 5a, 및 도 5b는 본 발명의 일 실시예에 따른, 원발성 또는 속발성 대장암 세포주에 대한 고시폴 및/또는 펜포르민 투여의 ATP 합성 저해 효과를 나타낸 도이다. 도 5a 및 도 5b에서 C는 대조군(control), G는 고시폴(gossypol), P는 펜포르민(phenformin), G+P는 고시폴과 펜포르민의 병용 투여를 나타낸다.4A, 4B, 5A, and 5B are diagrams showing the ATP synthesis inhibitory effect of administration of gosipol and/or phenformin to primary or secondary colorectal cancer cell lines according to an embodiment of the present invention. In FIGS. 5A and 5B , C denotes a control, G denotes gossypol, P denotes phenformin, and G+P denotes the combined administration of gossypol and phenformin.
도 6a, 및 도 6b는 본 발명의 일 실시예에 따른, 원발성 또는 속발성 대장암 세포주에 대한 고시폴 및/또는 펜포르민 투여의 젖산 발생 결과를 확인한 도이다. 도 6a 및 도 6b에서 C는 대조군으로서의 고시폴(gossypol), P는 펜포르민(phenformin), G+P는 고시폴과 펜포르민의 병용 투여를 나타낸다.6A and 6B are diagrams confirming the lactic acid generation results of administration of gossypol and/or phenformin to primary or secondary colorectal cancer cell lines according to an embodiment of the present invention. In FIGS. 6A and 6B , C denotes gossypol as a control, P denotes phenformin, and G+P denotes co-administration of gossypol and phenformin.
본 발명의 발명자들은 원발성 대장암 세포주, 및 속발성 대장암 세포주에서 고시폴 및/또는 펜포르민 효과를 확인한 결과, 원발성 대장암에 비해서 속발성 대장암에서 고시폴+펜포르민 병용 투여시 암세포 사멸 촉진 효과, 미토콘드리아 내 ATP 합성 억제 효과, 및 젖산 발생 억제 효과가 현저함을 확인하였다. 이로부터 고시폴+펜포르민 병용 투여에 의해 속발성 대장암을 효과적으로 치료할 수 있음을 알 수 있었다.As a result of confirming the effect of gosipol and/or phenformin in primary colorectal cancer cell lines and secondary colorectal cancer cell lines, the present inventors promote cancer cell death when administered in combination with gosipol + phenformin in secondary colon cancer compared to primary colorectal cancer It was confirmed that the effect, the mitochondrial ATP synthesis inhibitory effect, and the lactic acid generation inhibitory effect were significant. From this, it was found that secondary colorectal cancer could be effectively treated by the combined administration of gosipol + phenformin.
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 보다 구체적으로 설명하기 위한 것으로서, 본 발명의 요지에 따라 본 발명의 범위가 이들 실시예에 의해 제한되지 않는다는 것은 당업계에서 통상의 지식을 가진 자에게 있어서 자명할 것이다.Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention in more detail, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention. .
실시예 1: 암세포의 배양Example 1: Cultivation of cancer cells
본 발명의 발명자들은 한국 세포주 은행(Korea Cell Line Bank; KCLB), 또는 미국 세포주 은행(American Type Culture Collection; ATCC)으로부터 원발성 대장암 세포주(SW480), 및 속발성 대장암 세포주(LOVO, 및 SNU407)을 수득하여, KCLB, 또는 ATCC의 가이드에 따라 배양하였다. 구체적인 세포주 정보는 하기 표 1에 기재하였다.The inventors of the present invention obtained primary colorectal cancer cell lines (SW480), and secondary colorectal cancer cell lines (LOVO, and SNU407) from Korea Cell Line Bank (KCLB), or American Type Culture Collection (ATCC). obtained and cultured according to the guidelines of KCLB or ATCC. Specific cell line information is shown in Table 1 below.
Figure PCTKR2020017417-appb-T000001
Figure PCTKR2020017417-appb-T000001
실시예 2. 대장암 세포주에서 고시폴 및/또는 펜포르민 처리의 암 치료 효과 확인Example 2. Confirmation of cancer treatment effect of gosipol and/or phenformin treatment in colorectal cancer cell lines
실시예 1의 대장암 세포주에 고시폴을 단독 투여하거나, 또는 펜포르민과 병용 투여하여 항암 치료 결과를 확인하였다. 보다 구체적으로, 각각의 세포를 96-웰 플레이트에 1×104 cell/well 농도로 분주하고, 37℃, 5% CO2 환경에서 하룻밤 배양하였다. 이후, 고시폴을 0 내지 16μM의 농도로 단독 투여하거나 또는 10μM 또는 100μM의 펜포르민과 병용 투여하고, 37℃, 5% CO2 환경에서 추가로 24시간 배양하였다. 세포배양액 100㎕와 세포생존율 분석 키트(Cell Counting Kit-8)의 반응버퍼 10㎕를 혼합하여 96웰 플레이트에 넣고 실온에서 1시간 반응시킨 후에 분광광도계를 이용하여 450nm 파장에서 흡광도 측정하였다(Synergy HTX Multi-Reader, BioTek, Winooski, VT, USA). 동시에 각 시료의 세포 수를 세포생존율 분석 키트(Cell Counting Kit-8)로 측정하고, 측정값은 음성대조군의 세포 수에 대비하여 각 시료에서의 동일한 세포 수 대비 측정값이 비교될 수 있도록 백분율로 산출하였다. 모든 실험은 3회를 반복하여 평균하여, 그 결과를 도 1에 나타내었다.The results of anticancer treatment were confirmed by administering gosipol alone or in combination with phenformin to the colorectal cancer cell line of Example 1. More specifically, each cell was aliquoted in a 96-well plate at a concentration of 1×10 4 cell/well, and cultured overnight at 37° C., 5% CO 2 environment. Thereafter, gosypol was administered alone at a concentration of 0 to 16 μM, or co-administered with 10 μM or 100 μM of phenformin, and further cultured for 24 hours at 37° C., 5% CO 2 environment. 100 μl of the cell culture solution and 10 μl of the reaction buffer of the cell viability assay kit (Cell Counting Kit-8) were mixed, put in a 96-well plate, and reacted at room temperature for 1 hour. Multi-Reader, BioTek, Winooski, VT, USA). At the same time, the number of cells in each sample is measured with a cell viability analysis kit (Cell Counting Kit-8), and the measured value is calculated as a percentage so that the measured value can be compared with the same number of cells in each sample compared to the number of cells in the negative control group. calculated. All experiments were repeated three times and averaged, and the results are shown in FIG. 1 .
실험 결과, 실험된 모든 농도 범위에서 고시폴 단독, 고시폴+10μM 펜포르민, 또는 고시폴+100μM 펜포르민 투여군 모두 암세포의 사멸 효과가 있었으나, LOVO 세포주에서 SW480 세포주보다 약물 투여 효과가 매우 현저하였다.As a result of the experiment, all of the groups administered with Gosipol alone, Gosipol + 10μM phenformin, or Gosipol + 100μM phenformin had an effect on the killing of cancer cells in all concentration ranges tested, but the drug administration effect was very significant in the LOVO cell line than in the SW480 cell line. did.
또한, 고시폴 1μM, 펜포르민 100μM, 또는 이의 병용을 투여한 경우의 세포생존율 결과를 도 2에 나타내었다. In addition, the cell viability results in the case of administering 1 μM of gosypol, 100 μM of phenformin, or a combination thereof are shown in FIG. 2 .
실험 결과, 원발성 대장암 세포주인 SW480은 병용 투여 효과가 없었으나(통계적으로 유의하지 않음), 속발성 대장암 세포주인 LOVO는 고시폴과 펜포르민의 병용 투여 효과가 있는 것을 알 수 있었다.As a result of the experiment, the primary colorectal cancer cell line, SW480, had no effect of co-administration (statistically insignificant), but the secondary colorectal cancer cell line, LOVO, had an effect of co-administration of gosipol and phenformin.
상기 세포생존율 실험에서 사멸한 세포를 제외하고 살아있는 세포만을 염색하여 광학현미경으로 촬영한 결과를 도 3에 나타내었다. 도 3의 사진으로부터 SW480 세포주와 LOVO 세포주에서의 세포사 효과 차이를 더욱 명확하게 확인할 수 있었다.In the cell viability experiment, only living cells were stained except for dead cells, and the results obtained by photographing with an optical microscope are shown in FIG. 3 . From the photo of FIG. 3, the difference in the cell death effect in the SW480 cell line and the LOVO cell line could be more clearly identified.
실시예 3. 대장암 세포주에서 고시폴 및/또는 펜포르민 처리의 ATP 합성 저해 효과 확인Example 3. Confirmation of ATP synthesis inhibitory effect of gosipol and/or phenformin treatment in colorectal cancer cell lines
원발성 대장암 세포주(SW480), 또는 속발성 대장암 세포주(LOVO)에 실시예 2와 동일한 방법으로 고시폴 및/또는 펜포르민을 약물 처리하여 미토콘드리아에서의 ATP 합성 저해를 확인하였다. 그 결과를 도 4와 도 5에 나타내었다.Inhibition of ATP synthesis in mitochondria was confirmed by drug-treating a primary colorectal cancer cell line (SW480), or a secondary colorectal cancer cell line (LOVO) in the same manner as in Example 2, and/or phenformin. The results are shown in FIGS. 4 and 5 .
실험 결과, 세포생존율 실험 결과와 동일하게, 실험된 모든 농도 범위에서 고시폴 단독, 고시폴+10μM 펜포르민, 또는 고시폴+100μM 펜포르민 투여군 모두 ATP 합성 저해 효과가 있었으나, LOVO 세포주에서 SW480 세포주보다 약물 투여 효과가 매우 현저하였다.As a result of the experiment, the cell viability test results were identical to the results of the cell viability experiment, and all of the groups administered with Gosipol alone, Gosipol + 10μM phenformin, or Gosypol + 100μM Fenformin had an ATP synthesis inhibitory effect, but in the LOVO cell line, SW480 The effect of drug administration was much more pronounced than that of the cell line.
고시폴 1μM, 펜포르민 100μM, 또는 이의 병용을 투여한 경우에는 원발성 대장암 세포주인 SW480은 병용 투여 효과가 없었으나(통계적으로 유의하지 않음), 속발성 대장암 세포주인 LOVO는 고시폴과 펜포르민의 병용 투여 효과가 매우 두드러졌다. 따라서 상기 실시예 2에서의 세포 사멸 효과가 미토콘드리아에서의 ATP 합성 저해에 의한 영향임을 알 수 있었다.When gosipol 1μM, phenformin 100μM, or a combination thereof was administered, the primary colorectal cancer cell line, SW480, had no effect (statistically insignificant), but the secondary colorectal cancer cell line, LOVO, had no effect. The effect of concomitant administration of min was very pronounced. Therefore, it can be seen that the apoptosis effect in Example 2 is the effect of ATP synthesis inhibition in mitochondria.
실시예 4. 고시폴 및/또는 펜포르민 처리된 대장암 세포주의 세포주기 분석Example 4. Cell cycle analysis of colorectal cancer cell lines treated with gossypol and/or phenformin
원발성 대장암 세포주(SW480), 또는 속발성 대장암 세포주(LOVO, 및 SNU407)에 실시예 2와 동일한 방법으로 고시폴 및/또는 펜포르민을 약물 처리한 후, 세포주기별 세포의 비율을 분석하였다. 보다 구체적으로, 각각의 세포를 PBS로 2회 세척하여 70% 에탄올/PBS로 고정한 후, 0.1% saponin이 함유된 PBS에 용해된 0.5 ㎎/㎖의 RNase로 처리하여 37℃에서 30분간 반응시킨 후, 20 μg/㎖의 PI로 4℃에서 30분간 염색하였다. 세포의 DNA content는 1×106 개의 세포를 FACSCalibur 유세포분석기와 CellQuest 소프트웨어로 분석하였다. 상기 결과를 하기 표 2 내지 표 4에 나타내었다.After the primary colorectal cancer cell line (SW480) or the secondary colorectal cancer cell line (LOVO, and SNU407) was treated with gosipol and/or phenformin in the same manner as in Example 2, the ratio of cells for each cell cycle was analyzed. More specifically, each cell was washed twice with PBS, fixed with 70% ethanol/PBS, treated with RNase of 0.5 mg/ml dissolved in PBS containing 0.1% saponin, and reacted at 37° C. for 30 minutes , and stained with PI of 20 μg/ml at 4°C for 30 min. The DNA content of cells was analyzed in 1×10 6 cells with a FACSCalibur flow cytometer and CellQuest software. The results are shown in Tables 2 to 4 below.
Figure PCTKR2020017417-appb-T000002
Figure PCTKR2020017417-appb-T000002
Figure PCTKR2020017417-appb-T000003
Figure PCTKR2020017417-appb-T000003
Figure PCTKR2020017417-appb-T000004
Figure PCTKR2020017417-appb-T000004
실험 결과, 원발성 대장암 세포주인 SW480에 비해서 속발성 대장암 세포주인 SNU407과 LOVO에서 고시폴+펜포르민 병용 투여에 의한 sub-G1 단계의 세포 비율이 현저하게 증가하여, 고시폴+펜포르민 병용 투여가 원발성 대장암보다 속발성 대장암에 미치는 효과가 현저함을 알 수 있었다.As a result of the experiment, compared to the primary colorectal cancer cell line SW480, the ratio of cells in the sub-G1 stage was significantly increased in SNU407 and LOVO, the secondary colorectal cancer cell lines, due to the combined administration of gosypol + phenformin, and the gosipol + phenformin combination It was found that the effect of administration on secondary colorectal cancer was more significant than that of primary colorectal cancer.
실시예 5. 고시폴 및/또는 펜포르민 처리된 대장암 세포주의 젖산 발생 확인Example 5. Confirmation of lactic acid generation of gosipol and/or phenformin-treated colorectal cancer cell lines
원발성 대장암 세포주(SW480), 또는 속발성 대장암 세포주(LOVO, 및 SNU407)에 실시예 2와 동일한 방법으로 고시폴 1μM, 및/또는 펜포르민 100μM을 약물 처리한 후, 젖산 발생 효과를 확인하였다. 보다 구체적으로, 각각의 세포를 96 또는 24-웰 플레이트에 각각 1Х104, 또는 3Х105 cells/well 농도로 파종하고, 하룻밤 배양한 후에, 약물을 투여하고, 24시간 추가로 배양하였다. 이후 세포 배양액을 DPBS(Dulbecco's Phosphate-Buffered Saline, Wellgene, Korea)로 희석한 희석액 50㎕와 젖산 어세이용 반응버퍼(Lactate assay reaction buffer, Promega, Madison, WI, USA) 50㎕를 혼합하여 96-웰 플레이트에 넣고, 실온에서 1시간 반응시킨 후에, 분광광도계(Synergy HTX Multi-Reader, BioTek, Winooski, Vermont, U.S)로 발광을 측정하였다. 동시에 각 시료의 세포 수를 세포생존율 분석 키트(Cell Counting Kit-8)로 측정하여, 음성대조군의 세포 수에 대비하여 각 시료에서 동일한 세포 수 대비 젖산 측정값이 비교될 수 있도록 산출하였다. 상기 결과를 도 6에 나타내었다. After the primary colorectal cancer cell line (SW480), or the secondary colorectal cancer cell line (LOVO, and SNU407) was treated with 1 μM of gosipol and/or 100 μM of phenformin in the same manner as in Example 2, the lactic acid generation effect was confirmed did. More specifically, each cell was seeded in a 96 or 24-well plate at a concentration of 1Х10 4 , or 3Х10 5 cells/well, respectively, and cultured overnight, then the drug was administered and further cultured for 24 hours. After that, 50 μl of the cell culture solution diluted with DPBS (Dulbecco's Phosphate-Buffered Saline, Wellgene, Korea) and 50 μl of lactate assay reaction buffer (Lactate assay reaction buffer, Promega, Madison, WI, USA) were mixed and mixed for 96-well After putting on a plate and reacting at room temperature for 1 hour, luminescence was measured with a spectrophotometer (Synergy HTX Multi-Reader, BioTek, Winooski, Vermont, US). At the same time, the number of cells in each sample was measured with a cell viability analysis kit (Cell Counting Kit-8), and the lactate measurement value compared to the same number of cells in each sample was calculated so that the number of cells in the negative control group could be compared. The results are shown in FIG. 6 .
실험 결과, 고시폴 단독 투여시의 젖산 발생율을 100 기준으로 환산하였을 때, 펜포르민 단독 투여시 젖산 발생율 대비 고시폴+펜포르민 병용 투여시 젖산 발생율이 SW480 약 30%, SNU407 약 41%로, 속발성 대장암에서 원발성 대장암보다 젖산 발생 감소 효과가 현저함을 알 수 있었다.As a result of the experiment, when the lactate generation rate when Gosipol alone was administered was converted to 100, the lactate generation rate when phenformin alone was administered compared to the lactate generation rate when Gosipol + Fenformin was administered in combination was SW480 about 30% and SNU407 about 41%. , it was found that the effect of reducing lactate generation in secondary colorectal cancer was more significant than in primary colorectal cancer.
상기 실시예 1 내지 5의 결과로부터, 원발성 대장암에 비해서 속발성 대장암에서 고시폴+펜포르민 병용 투여시 암세포 사멸 촉진 효과, 미토콘드리아 내 ATP 합성 억제 효과, 및 젖산 발생 억제 효과가 현저함을 확인하였다. 이로부터 고시폴+펜포르민 병용 투여에 의해 속발성 대장암을 효과적으로 치료할 수 있음을 알 수 있었다.From the results of Examples 1 to 5, it was confirmed that the cancer cell death promoting effect, mitochondrial ATP synthesis inhibitory effect, and lactic acid generation inhibitory effect were remarkable when the gosipol + phenformin co-administration was administered in the secondary colorectal cancer compared to the primary colorectal cancer. did. From this, it was found that secondary colorectal cancer could be effectively treated by the combined administration of gosipol + phenformin.
이상으로 본 발명의 특정한 부분을 상세히 기술하였는 바, 당업계의 통상의 지식을 가진 자에게 있어서 이러한 구체적인 기술은 단지 바람직한 구현예일 뿐이며, 이에 본 발명의 범위가 제한되는 것이 아닌 점은 명백하다. 따라서 본 발명의 실질적인 범위는 청구범위에 기재된 본 발명의 기술적 사상을 벗어나지 않는 범위 내에서 다양한 수정 및 변형이 가능하다는 것은 당 기술분야의 통상의 지식을 가진 자에게는 자명할 것이다.As described above in detail a specific part of the present invention, for those of ordinary skill in the art, this specific description is only a preferred embodiment, and it is clear that the scope of the present invention is not limited thereto. Accordingly, it will be apparent to those of ordinary skill in the art that various modifications and variations are possible within the scope of the present invention as described in the claims.
속발성 암은 다른 부위에서 처음 발생한 암이 전이되어 새로운 장기로 전이한 이차성 암을 의미하는 것으로서, 원발성 암에 비하여 예후가 불량하다.Secondary cancer refers to secondary cancer that has metastasized to a new organ by first metastasizing cancer from another site, and has a poorer prognosis than primary cancer.
본 발명의 약학조성물은 원발성 암에 비하여 특이적으로 속발성 암에서 치료 효과가 현저하므로, 치료가 어렵고 예후가 불량한 속발성 암의 치료에 크게 이용될 것으로 기대된다.Since the pharmaceutical composition of the present invention has a significant therapeutic effect in secondary cancer specifically compared to primary cancer, it is expected to be widely used in the treatment of secondary cancer, which is difficult to treat and has a poor prognosis.

Claims (17)

  1. 알데히드 억제제(aldehyde inhibitor) 및 비구아나이드(biguanide) 계열 화합물을 포함하는 속발성 암의 예방 또는 치료용 약학조성물.A pharmaceutical composition for preventing or treating secondary cancer, including an aldehyde inhibitor and a biguanide-based compound.
  2. 제 1항에 있어서,The method of claim 1,
    상기 알데히드 억제제는 고시폴(Gossypol)인 것을 특징으로 하는 약학조성물. The aldehyde inhibitor is a pharmaceutical composition, characterized in that Gossypol (Gossypol).
  3. 제 1항에 있어서,The method of claim 1,
    상기 비구아나이드 계열 화합물은 펜포르민(Phenformin)인 것을 특징으로 하는 약학조성물. The biguanide-based compound is a pharmaceutical composition, characterized in that phenformin (Phenformin).
  4. 제 1항에 있어서, The method of claim 1,
    상기 알데히드 억제제와 비구아나이드 계열 화합물은 1:1~100의 중량비로 포함되는 것을 특징으로 하는, 약학조성물. The aldehyde inhibitor and the biguanide-based compound are characterized in that included in a weight ratio of 1:1 to 100, a pharmaceutical composition.
  5. 제 4항에 있어서, 5. The method of claim 4,
    상기 알데히드 억제제와 비구아나이드 계열 화합물은 1:2~20의 중량비로 포함되는 것을 특징으로 하는, 약학조성물.The aldehyde inhibitor and the biguanide-based compound are characterized in that included in a weight ratio of 1:2-20, a pharmaceutical composition.
  6. 제 1항에 있어서,The method of claim 1,
    상기 알데히드 억제제는 0.5 내지 50 μM의 양으로 포함되는 것을 특징으로 하는, 약학조성물. The aldehyde inhibitor is characterized in that included in an amount of 0.5 to 50 μM, a pharmaceutical composition.
  7. 제 1항에 있어서,The method of claim 1,
    상기 비구아나이드 계열 화합물은 10 내지 1000 μM의 양으로 포함되는 것을 특징으로 하는, 약학조성물. The biguanide-based compound is characterized in that it is included in an amount of 10 to 1000 μM, a pharmaceutical composition.
  8. 제 1항에 있어서,The method of claim 1,
    상기 속발성 암은 속발성 대장암인 것을 특징으로 하는, 약학조성물.The secondary cancer is characterized in that the secondary colorectal cancer, pharmaceutical composition.
  9. 제 8항에 있어서,9. The method of claim 8,
    상기 속발성 암은 주변 장기로부터 침윤되어나, 또는 혈관 또는 림프절로부터 원격 전이된 것을 특징으로 하는 약학조성물.The secondary cancer is infiltrated from the surrounding organs, or a pharmaceutical composition, characterized in that the distant metastasis from blood vessels or lymph nodes.
  10. 제 1항에 있어서,The method of claim 1,
    상기 암은 암 줄기세포를 포함하는 것을 특징으로 하는, 약학조성물.The cancer is characterized in that it comprises cancer stem cells, a pharmaceutical composition.
  11. 알데히드 억제제(aldehyde inhibitor) 및 비구아나이드(biguanide) 계열 화합물을 혼합하는 단계;를 포함하는, 약학조성물의 제조 방법.Mixing the aldehyde inhibitor (aldehyde inhibitor) and the biguanide (biguanide)-based compound; comprising, a method for preparing a pharmaceutical composition.
  12. 알데히드 억제제(aldehyde inhibitor) 및 비구아나이드(biguanide) 계열 화합물을 유효성분으로 포함하는 약학조성물로 속발성 암을 예방 또는 치료하는 방법.A method for preventing or treating secondary cancer with a pharmaceutical composition comprising an aldehyde inhibitor and a biguanide-based compound as an active ingredient.
  13. 제 12항에 있어서,13. The method of claim 12,
    상기 알데히드 억제제는 고시폴(Gossypol)인 것을 특징으로 하는, 방법.The method, characterized in that the aldehyde inhibitor is Gossypol (Gossypol).
  14. 제 12항에 있어서,13. The method of claim 12,
    상기 비구아나이드 계열 화합물은 펜포르민(Phenformin)인 것을 특징으로 하는, 방법.The method, characterized in that the biguanide-based compound is phenformin (Phenformin).
  15. 알데히드 억제제(aldehyde inhibitor) 및 비구아나이드(biguanide) 계열 화합물의 속발성 암 예방 또는 치료 용도.Use of aldehyde inhibitors and biguanide-based compounds for the prevention or treatment of secondary cancer.
  16. 알데히드 억제제(aldehyde inhibitor) 및 비구아나이드(biguanide) 계열 화합물을 유효성분으로 포함하는 약학조성물의 속발성 암 예방 또는 치료 용도.Secondary cancer prevention or treatment use of a pharmaceutical composition comprising an aldehyde inhibitor and a biguanide-based compound as an active ingredient.
  17. 알데히드 억제제(aldehyde inhibitor) 및 비구아나이드(biguanide) 계열 화합물을 유효성분으로 포함하는, 속발성 암 예방 또는 치료에 사용하기 위한 조성물.A composition for use in the prevention or treatment of secondary cancer, comprising an aldehyde inhibitor and a biguanide-based compound as an active ingredient.
PCT/KR2020/017417 2019-12-02 2020-12-02 Pharmaceutical composition for prevention and treatment of secondary cancer, comprising aldehyde inhibitor and biguanide-based compound WO2021112541A1 (en)

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Citations (5)

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KR20130019351A (en) * 2011-08-08 2013-02-26 한올바이오파마주식회사 N1-cyclic amine-n5-substituted phenyl biguanide derivatives, methods of preparing the same and pharmaceutical composition comprising the same
KR20160094861A (en) * 2015-02-02 2016-08-10 연세대학교 산학협력단 Pharmaceutical composition for inhibiting a growth of cancer stem cells comprising aldehyde inhibitor and biguanide compounds
KR20180054597A (en) * 2015-08-12 2018-05-24 시그모이드 파마 리미티드 Composition
WO2018155921A1 (en) * 2017-02-22 2018-08-30 국립암센터 Pharmaceutical composition for preventing and treating pancreatic cancer, containing gossypol and phenformin as active ingredients
KR101904893B1 (en) * 2016-02-18 2018-10-10 연세대학교 산학협력단 Pharmaceutical composition comprising polyphenol compound for cancer

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20130019351A (en) * 2011-08-08 2013-02-26 한올바이오파마주식회사 N1-cyclic amine-n5-substituted phenyl biguanide derivatives, methods of preparing the same and pharmaceutical composition comprising the same
KR20160094861A (en) * 2015-02-02 2016-08-10 연세대학교 산학협력단 Pharmaceutical composition for inhibiting a growth of cancer stem cells comprising aldehyde inhibitor and biguanide compounds
KR20180054597A (en) * 2015-08-12 2018-05-24 시그모이드 파마 리미티드 Composition
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