JP6484448B2 - Novel gem-dihydroperoxide compounds - Google Patents

Description

  The present invention relates to novel gem-dihydroperoxide compounds and uses thereof.

  A gem-dihydroperoxide compound, which is a peroxide, is a compound having a structure in which two hydroperoxide groups are bonded to the same atom. Synthetic intermediates such as trioxane, tetraoxane and endoperoxide, radical polymerization initiators, and epoxidation And is used as an oxidizing agent for sulfoxylation. In addition, gem-dihydroperoxide compounds are also attracting attention as synthetic raw materials for antimalarial drugs.

  Non-Patent Document 1 describes a method of synthesizing a gem-dihydroperoxide compound using oxygen and anthraquinone under irradiation with visible light.

L. Cui et al. Green Chem. , (2011) 13, 2347-2350

  An object of the present invention is to provide a novel gem-dihydroperoxide compound.

  The present inventors have found that a novel gem-dihydroperoxide compound has high antioxidant activity and anticancer activity, and have completed the present invention.

  According to the present invention, novel gem-dihydroperoxide compounds are provided. Since the gem-dihydroperoxide compound has high antioxidant activity and anticancer activity, it is useful as an antioxidant and an anticancer agent.

The ESR spectrum measured using DMSO (A), the compound (B, C) which concerns on this invention, or a comparison compound (D) is shown. The ratio of living cells when leukemia cells are treated with DMSO, a compound according to the present invention, or a comparative compound is shown. The time-dependent change of the ratio of a living cell at the time of processing a leukemic cell with DMSO or the compound concerning this invention is shown. It is an electron microscope image of leukemia cells treated with DMSO or a compound according to the present invention. It is a Hoechst33342 dyeing | staining image of the leukemia cell processed with DMSO or the compound concerning this invention. The percentage of cells undergoing apoptosis in leukemia cells treated with DMSO or a compound according to the present invention is shown. The result of having analyzed the apoptosis related protein in the leukemia cell processed with DMSO or the compound concerning this invention by the Western blot is shown. The ratio of living cells when human normal lymphocytes are treated with DMSO or a compound according to the present invention is shown.

  Embodiments of the present invention will be described below. In addition, this invention is not limited only to the following embodiment.

  In this specification, “X to Y” indicating a range means “X or more and Y or less”. Unless otherwise specified, measurement of operation and physical properties is performed under conditions of room temperature (20 to 25 ° C.) / Relative humidity 40 to 50%.

(Gem-dihydroperoxide compound)
In the first aspect of the present invention, a novel compound represented by the following formula (1) or a physiologically acceptable salt thereof (hereinafter referred to as a compound represented by the following formula (1) or a physiologically acceptable salt thereof) Simply referred to as “compounds according to the invention”).

However, in the above formula (1), the R ¹ is a substituted or unsubstituted, linear, branched or a saturated hydrocarbon group having 1 to 4 carbon atoms alicyclic structure; R ² is a substituted or unsubstituted , A saturated hydrocarbon group having 7 to 13 carbon atoms including a straight chain, branched chain or alicyclic structure; the total carbon number of R ¹ and R ² is 11 to 14.

  Specific examples of the saturated hydrocarbon group having 1 to 4 carbon atoms include methyl group, ethyl group, n-propyl group, isopropyl group, cyclopropyl group, cyclopropylmethyl group, 2-methylcyclopropyl group, Examples include n-butyl group, sec-butyl group, tert-butyl group, and cyclobutyl group.

  Specific examples of the saturated hydrocarbon group having 7 to 13 carbon atoms include alkyl groups such as heptyl group, octyl group, nonyl group, decyl group, undecyl group, dodecyl group and tridecyl group, as well as alkyl substitution. Alternatively, those having an alicyclic structure such as an unsubstituted monocyclic ring (for example, a cycloalkyl group), a polycyclic ring (for example, a bicyclohexyl group), or a condensed ring (decahydronaphthyl group) can be mentioned. Examples of the saturated hydrocarbon group containing an alkyl-substituted alicyclic structure include a methylcyclohexyl group, an ethylcyclohexyl group, a cyclohexylethyl group, and the like.

  The compound represented by the above formula (1) is a kind of gem-dihydroperoxide compound having two dihydroperoxy groups on a single carbon atom. The gem-dihydroperoxide compounds are used as synthetic intermediates such as trioxane, tetraoxane and endoperoxide, radical polymerization initiators, and oxidizing agents for epoxidation and sulfoxylation.

In the above formula (1), R ¹ is preferably an alkyl group having 1 to 2 carbon atoms from the viewpoint of antioxidant activity and anticancer activity.

In the above formula (1), in view of the anti-cancer activity, it is preferable that the carbon number of ^{R 2} is 8 to 13, more preferably 9-11. R ² is preferably a linear or branched alkyl group having any one of the above carbon numbers.

Examples of the substituent for R ¹ and R ² include a halogen atom, a hydroxyl group, an acyl group having 2 to 3 carbon atoms, and an alkoxy group having 1 to 3 carbon atoms.

  Examples of the halogen atom include a fluorine atom, a chlorine atom, a bromine atom, and an iodine atom.

  Examples of the acyl group include an acetyl group and a propionyl group.

  Examples of the alkoxy group include a methoxy group, an ethoxy group, and a propoxy group.

The total number of carbon atoms of R ¹ and R ² may be 11 to 14, but is preferably 11 to 13 and more preferably 11 to 12 from the viewpoint of anticancer activity.

From the standpoint of antioxidant activity and anticancer activity, a preferred embodiment of the present invention, in the above-mentioned formula (1), R ¹ is alkyl group having 1 or 2 carbon atoms; R ² is a substituted or unsubstituted , A saturated hydrocarbon group having 9 to 11 carbon atoms including a linear, branched or alicyclic structure, and the total carbon number of R ¹ and R ² is 11 to 12.

From the standpoint of antioxidant activity and anticancer activity, another more preferred embodiment of the present invention, in the above-mentioned formula (1), R ¹ is alkyl group having 1 or 2 carbon atoms; R ² is a substituted or unsubstituted It is a substituted alkyl group having 9 to 11 carbon atoms, and the total number of carbon atoms of R ¹ and R ² is 11 to 12.

  Specific examples of the compound according to the present invention are illustrated below, but the present invention is not limited thereto. In the following, the number immediately before “AC” indicates the number of carbon atoms contained in the compound, and the number immediately before “O” indicates the position of carbon to which two hydroperoxide groups are connected.

  As used herein, the term “physiologically acceptable salt” refers to a salt that does not cause undesirable reactions such as excessive toxicity, irritation, allergic reactions and other complications when administered to mammals, particularly humans. For example, salts of suitable cations (sodium, calcium, potassium, magnesium, ammonium, etc.). The peroxide anion from which the hydrogen atom in the hydroperoxide group of the above formula (1) is eliminated and the cation can form an ion pair.

  The compound represented by Formula (1) can be obtained using a conventionally known method as a method for synthesizing a gem-dihydroperoxide compound. For example, N.I. Tada et al. , Adv. Synth. Catal. (2010) 352, 2383-2386, N.A. Tada et al. , Chem. Commun. , (2010) 46, 1772-1774, or non-patent document 1 or the like. More specifically, a ketone is used as a raw material, and the carbonyl group of the raw material ketone may be oxidized into two hydroperoxide groups using hydrogen peroxide or the like.

The ketone used as the raw material may be selected according to the structure of the target gem-dihydroperoxide compound (for example, the number of carbon atoms and the position of the dihydroperoxide). Since the carbon to which the carbonyl group in the ketone is linked becomes the carbon to which the hydroperoxide group is linked in the compound of the formula (1), the position of the carbonyl group to be used is appropriate according to the length of the target R ¹ or R ^2. A suitable ketone is selected as a raw material. For example, when 3-dodecanone is used as a raw material, a compound represented by formula (1) (compound 2 above) in which R ¹ has 2 carbon atoms and R ² has 9 carbon atoms can be obtained.

  As a ketone used as a raw material, those having 12 to 15 carbon atoms are used. As the ketone used as a raw material, a linear or branched chain having 12 to 13 carbon atoms is preferably used. Examples of the substituent of the ketone group include those described above in the compound according to the present invention.

  More specifically, examples of the raw material ketone include 2-dodecanone, 3-dodecanone, 4-dodecanone, 5-dodecanone, 2-tridecanone, 3-tridecanone, 4-tridecanone, 5-tridecanone, 2-tetradecanone, 3 -Tetradecanone, 4-tetradecanone, 5-tetradecanone, 2-pentadecanone, 3-pentadecanone, 4-pentadecanone, 5-pentadecanone, 2-methyl-3-undecanone, 5-ethyl-3-decanone, 2-methyl-5-ethyl -3-decanone, 1- (1-methylcyclohexyl) ethyl ethyl ketone and the like.

  The above raw material ketone is oxidized in a solvent to synthesize a gem-dihydroperoxide compound. The solvent to be used is not particularly limited. For example, polar solvents such as methanol, ethanol, 2-propanol, tert-butanol, acetone, acetonitrile, 1,2-dimethoxyethane, hexane, methylene chloride, Nonpolar solvents such as diethyl ether, ethyl acetate, toluene can be used. These solvents may be used alone or in combination of two or more.

  The amount of the raw material ketone added to the solvent is not particularly limited, and is, for example, 0.001 to 10 mol / L, and preferably 0.01 to 1 mol / L.

  1 to 10 molar equivalents of hydrogen peroxide or the like is added to the raw material ketone dissolved in the solvent to oxidize the carbonyl group of the raw material ketone to dihydroperoxide. At this time, a catalyst such as iodine may be added as necessary. The addition amount of the catalyst is not particularly limited, and is, for example, 0.0001 to 0.5 mol with respect to 1 mol of the raw material ketone.

  The oxidation reaction of the raw material ketone dissolved in the solvent is, for example, N.I. Tada et al. , Adv. Synth. Catal. , (2010) 352, 2383-2386, Non-Patent Document 1, etc., may be performed by photooxidation in an oxidizing gas atmosphere such as oxygen. More specifically, a high pressure mercury lamp, an electrodeless lamp, a xenon lamp, a metal halide lamp, or the like is used as a light source, and an oxidation reaction is performed by irradiating the raw material ketone solution with ultraviolet rays or visible light. In this case, a photosensitizer such as an anthracene compound, anthraquinone compound, pyrene compound, redox compound, azo compound, or diazo compound may be appropriately added as necessary.

  The temperature of the oxidation reaction may be arbitrarily set, and is, for example, 10 to 50 ° C., preferably 20 to 40 ° C. The reaction time is not particularly limited, and is, for example, 0.1 to 50 hours, preferably 5 to 30 hours.

  The compound according to the present invention obtained by the oxidation reaction may be separated and purified by a conventionally known method. Examples of the separation and purification method include, but are not limited to, one means such as filtration, extraction (for example, ether extraction), distribution, crystallization, and chromatography (for example, medium pressure silica gel column chromatography). Or it can carry out in combination of 2 or more.

  The structure of the obtained compound can be analyzed by means known to those skilled in the art, such as NMR, mass spectrometry, and spectroscopic measurement. More specifically, it can be analyzed by the means described in the examples.

(Antioxidant)
The generation of active oxygen and free radicals in the living body is considered to be related to the onset of diseases such as inflammatory diseases, and the development of components having antioxidant activity in the field of pharmaceuticals and cosmetics is underway. . The inventors have found that the compounds according to the invention have a high antioxidant activity. Therefore, in the second aspect of the present invention, there is provided an antioxidant comprising as an active ingredient at least one selected from the compound represented by formula (1) or a physiologically acceptable salt thereof.

  A compound having a hydroperoxide group generally has an oxidation activity. The gem-dihydroperoxide compounds according to the invention also have hydroperoxide groups, but surprisingly have a high antioxidant activity.

  The compound according to the present invention can effectively eliminate active oxygen species such as superoxide anion and hydroxyl radical.

  In a living body that obtains energy using oxygen, free radicals such as reactive oxygen species (ROS) and lipid peroxide are continuously produced. ROS and free radicals are highly reactive, and modify functional molecules such as proteins, lipids, and DNA to cause cell dysfunction. Although cells have a defense mechanism against ROS and free radicals, it is thought that when excessive ROS accumulates, oxidative stress occurs, and the onset and symptoms of diseases associated with oxidative stress such as neurodegenerative diseases progress. The antioxidant according to the second aspect of the present invention can effectively erase active oxygen species and free radicals in a living body as an active oxygen scavenger and / or a free radical scavenger. Therefore, the antioxidant according to the second aspect of the present invention prevents and / or treats neurodegenerative diseases, vascular disorders, heart diseases, inflammatory diseases, eye diseases and / or skin diseases in which oxidative stress is involved. In particular, it can be suitably used for inflammatory diseases.

  In one embodiment of the present invention, there is provided a prophylactic and / or therapeutic agent for oxidative stress diseases comprising, as an active ingredient, one or more selected from the compounds according to the present invention and physiologically acceptable salts thereof. . In one embodiment, the oxidative stress disease is selected from the group consisting of neurodegenerative disease, vascular disorder, heart disease, inflammatory disease, eye disease, and skin disease. In the present specification, “a prophylactic and / or therapeutic agent for oxidative stress disease” is also referred to as “antioxidative stress agent”.

  Examples of neurodegenerative diseases include Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, polyglutamine disease, and prion disease. Examples of the vascular disorder include stroke, cerebral infarction, transient ischemic attack, vasculitis and the like. Examples of heart diseases include angina and myocardial infarction. Examples of inflammatory diseases include ulcerative colitis, Crohn's disease, asthma, acute respiratory distress syndrome, fulminant hepatitis, irritable lung disease, rheumatoid arthritis, allergic rhinitis, type I diabetes, and Sjögren's syndrome. Examples of eye diseases include retinopathy, cataract, glaucoma, age-related macular degeneration, and the like. Examples of skin diseases include sunlight dermatitis, eczema, atopic dermatitis, allergic contact dermatitis, and urticaria.

  When the antioxidant or antioxidant stress agent according to the present invention is administered as a pharmaceutical, the subjects to whom the administration is performed include humans and non-human animals. Non-human animals include vertebrates, eg, non-human primates (especially higher primates), dogs, rodents (eg, mice or rats), mammals such as guinea pigs, cats and rabbits, and birds Non-mammals such as amphibians, reptiles and the like. In one embodiment, the subject is a human. In another embodiment, the subject is a non-human animal or animal suitable as a disease model.

  In the present specification, “containing as an active ingredient” means an amount sufficient to exhibit a desired activity (antioxidant activity, preventive or therapeutic activity for oxidative stress disease, anticancer activity, etc.) (ie, effective Means to contain by quantity). Therefore, the antioxidant and antioxidant stress agent according to the present invention may contain only the compound according to the present invention, but as long as the desired activity is not impaired, a pharmaceutically acceptable carrier and other additives You may comprise a composition with an agent etc. Moreover, the antioxidant and the antioxidant stress agent according to the present invention can be mixed with additives such as excipients and used in a form suitable for parenteral administration, oral administration or external administration. For example, according to a preferred embodiment of the antioxidant or the antioxidant stress agent according to the present invention, a skin external preparation composition comprising the above-described compound according to the present invention and a pharmaceutically acceptable carrier is provided. Is done. When the skin external preparation composition is a pharmaceutical product, it can be used for the prevention and / or treatment of a disease or condition associated with oxidative stress in skin tissue. Moreover, when the said skin external preparation composition is cosmetics, it can be used for prevention and / or improvement of the disease or pathological condition which the oxidative stress in a skin tissue is related.

  When the antioxidant or antioxidant stress agent according to the present invention is used as a medicament, the dosage varies depending on the target disease, administration subject, administration route and the like. For example, since the dose can easily vary depending on conditions such as the body weight of the subject person, the dosage can be appropriately selected by those skilled in the art, but generally, the dry weight is 0.001 to 1000 mg / day / kg body weight. A range is desirable.

  For use in medicine or cosmetics, an effective amount of a compound according to the invention may be formulated with one or more pharmaceutically acceptable carriers (additives) and / or diluents. As described in detail below, the antioxidants and antioxidant stress agents according to the present invention can be formulated for administration in solid or liquid form. Examples of the orally administered agent include liquid medicine (aqueous solution or non-aqueous solution or suspension), tablet, bolus, powder medicine, granule, and paste. The parenteral agent can be formulated into, for example, a subcutaneous, intraperitoneal, intramuscular or intravenous injection, or a dosage form for intravaginal or rectal administration. When it is used as an external preparation for skin, the form is not particularly limited, and pharmaceutical forms such as sprays, ointments, pastes, creams, lotions, gels, solutions, patches; skin lotions, cosmetic emulsions, cosmetic creams, cosmetic gels, Examples are cosmetic forms of skin care products or makeup products such as cosmetic liquids, packs, foundations, lipsticks, lip balms, lip glosses, facial cleansers, body soaps, hand creams, shampoos, rinses, hair styling agents and the like. In particular, in the case of a cosmetic form, a lotion, a cosmetic gel, a cosmetic emulsion, a cosmetic cream, and a cosmetic liquid are preferable, and a lotion, a cosmetic emulsion, and a cosmetic cream are more preferable because they can be applied to a wide range of sites. .

  “Pharmaceutically acceptable carrier” means a liquid or solid pharmaceutical agent involved in transporting or transporting a composition according to the present invention from one organ or part of the body to another organ or part of the body. It means an acceptable filler, diluent, excipient, solvent, encapsulating material, excipient, or composition thereof. Pharmaceutically acceptable carriers include, for example, sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and derivatives thereof such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; tragacanth; gelatin Talc; excipients such as cocoa butter and suppository wax; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols such as propylene glycol; glycerin, sorbitol, mannitol and Polyols such as polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; like magnesium hydroxide and aluminum hydroxide Do buffers; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol; phosphate buffer solution and the like.

  Antioxidants and antioxidant stress agents according to the present invention may be present in the composition together with other pharmaceutically acceptable antioxidants (antioxidants). Examples of other pharmaceutically acceptable antioxidants include, for example: water-soluble antioxidants such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium disulfite, sodium sulfite and the like; palmitic acid Oil-soluble antioxidants such as ascorbyl, butylhydroxyanisole (BHA), butylhydroxytoluene (BHT), lecithin, propyl gallate, α-tocopherol and the like; and citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, A metal chelating agent such as phosphoric acid can also be included as required.

  In addition, wetting agents such as sodium lauryl sulfate and magnesium stearate, emulsifiers, lubricants, colorants, release agents, coatings, sweeteners, fragrances, preservatives and antioxidants may also be present in the composition. Good.

  The antioxidant and antioxidant stress agent according to the present invention formulated in a form suitable for parenteral administration, together with the compound according to the present invention, one or more pharmaceutically acceptable solvents, dispersants, emulsifiers, Antioxidants, buffers, bacteriostatic agents, isotonic agents, and / or suspending agents may be included. When used for parenteral administration, the compound according to the present invention is purified water, an appropriate buffer solution such as a phosphate buffer, physiological saline, Ringer solution (Ringer solution), physiological salt solution such as a lock solution, ethanol, etc. Can be formulated as an aqueous solution, non-aqueous solution, suspension, liposome or emulsion, suitably combined with glycerin and surfactants. For example, it is formulated as an aqueous solution for injection and administered intravenously, intraperitoneally, subcutaneously, intramuscularly and the like. At this time, the preparation preferably has a physiological pH, preferably a pH within the range of 6-8. In addition, liquid preparations such as lotions, suspensions and emulsions, semi-solid preparations such as gels, creams and ointments, powders, powders (powder), solid preparations for use in preparations, or external preparations such as patches As such, it may be administered percutaneously to the target site and its surrounding sites. Furthermore, it can be administered as a suppository using a suppository base. Among the above, preferred preparations, administration forms, and the like can be selected by the doctor in charge. Semi-solid preparations such as lotions, creams or ointments comprise compounds according to the invention, oils, lanolin, petrolatum, paraffin, wax, plaster, resins, glycols, higher alcohols, glycerin, water, emulsifiers and suspensions. It is obtained by appropriately mixing with one or more selected from the group consisting of agents.

  Antioxidants and antioxidant stress agents may include adjuvants such as preservatives, wetting agents, emulsifiers and dispersants and various antibacterial and antifungal agents such as parabens, chlorobutanol, phenol sorbate, etc. Good. Isotonic agents such as sugar and sodium chloride can also be included in the composition. Furthermore, by using agents that delay absorption, such as aluminum monostearate and gelatin, a formulation with sustained absorption can be obtained.

  In one embodiment of the present invention, there is provided a method for preventing and / or treating an oxidative stress disease using the novel compound represented by the above formula (1) or a physiologically acceptable salt thereof.

  In another embodiment of the present invention, there is provided use of a novel compound represented by the above formula (1) or a physiologically acceptable salt thereof in the manufacture of a preventive and / or therapeutic agent for oxidative stress disease.

  In these embodiments, the oxidative stress disease can be exemplified by the above-mentioned neurodegenerative diseases, vascular disorders, heart diseases, inflammatory diseases, eye diseases and skin diseases, etc. Preferably used.

(Anti-cancer agent)
Malignant neoplasms (cancers) have been ranked first in the age-adjusted mortality rate by major cause of death in the “Demographic Statistics” of the Ministry of Health, Labor and Welfare from 1981 to the present, and the death rate from cancer Has not shown a downward trend until recently, but has been flat or slightly increasing. For this reason, many studies have been conducted to develop treatment methods suitable for cancer treatment and symptom reduction. In the field of chemical therapy, antibiotics, antimetabolites, alkylating agents, hormone agents, and the like have been found as effective anticancer agents for cancer cells. However, these anticancer drugs not only attack cancer cells but also act on normal cells, causing problems that cause side effects such as vomiting, nausea, loss of appetite, and hair loss. Development of a novel therapeutic agent with few side effects is desired. The present inventors have found that the compounds according to the present invention have the advantages of having high anticancer activity and extremely low cytotoxicity against normal cells. Therefore, in the third aspect of the present invention, there is provided an anticancer agent comprising, as an active ingredient, one or more selected from the compound represented by formula (1) or a physiologically acceptable salt thereof.

  The mechanism by which the compound according to the present invention exhibits anticancer activity is considered as follows. That is, it is presumed that the compound according to the present invention induces cell cycle arrest and apoptosis and exerts anticancer activity. On the other hand, it is considered that normal cells with a healthy cell membrane have low uptake of the compound according to the present invention and that it is difficult to induce cytotoxicity in normal cells that are not cancerous. In addition, the said content is estimation and does not restrict | limit the technical scope of this invention.

  The anticancer agent according to the present invention is administered to a subject as a medicament for the treatment, prevention, reduction, and / or treatment of cancer. Here, the cancer in which the anticancer agent according to the present invention is used is not particularly limited as long as the growth of cancer cells is suppressed. For example, colorectal cancer, stomach cancer, esophageal cancer, breast cancer. , Leukemia, lung cancer, prostate cancer, liver cancer, biliary tract cancer, spleen cancer, kidney cancer, bladder cancer, uterine cancer, ovarian cancer, testicular cancer, thyroid cancer, pancreatic cancer, brain tumor , Hematopoietic tumor, malignant melanoma and the like. The anticancer agent according to the present invention can be used particularly effectively for leukemia.

  The dose of the anticancer agent according to the present invention varies depending on the target disease, administration subject, administration route and the like. For example, since the dose can be easily changed depending on conditions such as the body weight of the subject person, the dosage can be appropriately selected by those skilled in the art. For example, the dry weight ranges from 0.001 to 1000 mg / day / kg body weight. It is desirable that

  The above-mentioned description regarding the dosage form, formulation and formulation of the antioxidant and the antioxidant stress agent, and the subject and the like is applied to the anticancer agent according to the present invention as appropriate.

  The anticancer agent according to the present invention may be used in combination with other anticancer agents as necessary. Other anticancer agents include, for example, fluorouracil, tegafur, tegafur / uracil combination agent, tegafur / gimeracil / oteracil potassium combination agent, doxyfluridine, capecitabine, carmofur, cytarabine, cytarabine ocphosphate, enocitabine, gemcitabine, azacitidine, Decitabine, floxuridine, ethynylcytidine, 6-mercaptopurine, fludarabine, pentostatin, nelarabine, 6-thioguanine, cladribine, clofarabine, methotrexate, pemetrexed, raltitrexed, nolatrexed, pralatrexate, trimethrexate, carbatrexate Cisplatin, carboplatin, nedaplatin, oxaliplatin, satraplatin, miriplatin, Baplatin, spiroplatin, tetraplatin, ormaplatin, iproplatin, cyclophosphamide, ifosfamide, busulfan, melphalan, nitrogen mustard, chlorambucil, glufosfamide, mafosfamide, estramustine, nimustine, ranimustine, carmustine, lomustine, semustine , Streptozocin, procarbazine, dacarbazine, temozolomide, thiotepa, hexamethylmelamine, trabectedin, apadicon, artretamine, bendamustine, mitractol, anthracycline antibiotics (e.g., doxorubicin, daunorubicin, pirarubicin, epirubicin, arubicin, arubicin, arubicin, arubicin Valrubicin, liposomal doxorubi , Pixantrone, and mitoxantrone), mitomycin C, bleomycin, pepromycin, actinomycin D, dinostatin stimamarer, topotecan, irinotecan, exatecan, nogitecan, etoposide, teniposide, zobuzoxan, vincristine, vinblastine, vinolsine, vinorelbine, vinorelbine Monomethyl auristatin E, epothilone B, eribulin, paclitaxel, docetaxel, cabazitaxel, tamoxifen, toremifene, raloxifene, fulvestrant, anastrozole, exemestane, letrozole, aminoglutethimide, formestane, borozole, methyltestosterone, medroxyprogesterone , Megestrol, guestnolone, mepithiostan, flutamide, Nilutamide, bicalutamide, finasteride, chlormadinone, estramustine, diethylstilbestrol, ethinylestradiol, phosfestol, polyestradiol phosphate, prednisolone, dexamethasone, mitotane, goserelin, leuprorelin, buserelin, triptorelin, bevacizumab, africeptumab , MV833, cetuximab, pegaptanib, pazopanib, CBO-P11, sunitinib, sorafenib, ranibizumab, batalanib, axitinib, zactima, NX1838, angiozyme, semaxanib, restaurtinib, TSU-68T 470, CP-547632, CPE-7055, KRN633, A E788, IMC-1211B, PTC-299, E7820, lenvatinib, marimastat, neobasstat, purinostat, metastat, BMS-275291, MMI270, S-3304, vitaxin, orotic acid carboxamidotriazole, thalidomide, genistein, interferon α , And interleukin 12 and the like.

  In one embodiment of the present invention, a method for preventing and / or treating cancer using the novel compound represented by the above formula (1) or a physiologically acceptable salt thereof is provided. The method according to the present embodiment can be performed in appropriate combination with other known treatment methods such as surgical treatment and radiation therapy.

  In another embodiment of the present invention, the use of a novel compound represented by the above formula (1) or a physiologically acceptable salt thereof in the manufacture of an anticancer agent is provided.

(Cosmetics)
In a fourth aspect of the present invention, there is provided a cosmetic comprising one or more selected from the compound represented by formula (1) or a physiologically acceptable salt thereof. Since the compound according to the present invention is excellent in antioxidant properties, it can effectively prevent oxidative deterioration of cosmetics and can protect the skin from oxidative damage and aging.

  At this time, the cosmetics (cosmetic composition) are in the form of lotion, cosmetic emulsion, cosmetic cream, cosmetic gel, cosmetic liquid, pack agent, foundation, lipstick, lip lip, lip gloss, facial cleanser, body. Skin care products such as soaps, hand creams, shampoos, rinses, hair styling products, and makeup products may be mentioned, but the invention is not particularly limited to these. Such a cosmetic (cosmetic composition) containing the compound according to the present invention can be produced using techniques known to those skilled in the art.

  Cosmetics (cosmetic compositions) containing the compounds according to the invention comprise physiologically acceptable carriers suitable for application to the skin, for example the carriers described for the abovementioned antioxidants. In order for the compound according to the present invention to be applied at an appropriate concentration and kept uniform, known diluents, dispersants, solvents and the like can be used as carriers. The carrier can be a solid, semi-solid, or liquid.

  Cosmetics containing the compounds according to the present invention include oils and fats such as animal fats and vegetable fats, waxes, paraffins, starches, tragacanths, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acids, talc and zinc oxides, or mixtures thereof. Excipients such as

  The compounding amount of the compound according to the present invention in these cosmetics (cosmetic compositions) is difficult to define uniformly depending on the type and state of the cosmetics, but is 0.01 to 80% by mass relative to the total weight of the cosmetics. More preferably, it is 0.1-20 mass%.

  The effects of the present invention will be described more specifically with reference to the following examples. However, the technical scope of the present invention is not limited only to the following examples.

<1. Production of gem-dihydroperoxide compound>
(Production Example 1)
Compound 2 (12AC-3O), which is a kind of the compound represented by formula (1), was obtained by the following method. The production method of Compound 2 (12AC-3O) is schematically shown below. In the above “12AC-3O”, the numerical value “12” indicates the number of carbon atoms contained in the compound, the numerical value “3” indicates the position of carbon to which two hydroperoxide groups are connected, and the compounds in Tables 1 to 3 below. The name is the same. The position of carbon to which the hydroperoxide group is connected is the position where the carbonyl group of the raw material ketone exists.

  Specifically, 3-dodecanone (110.6 mg, 0.6 mmol) and iodine (7.6 mg, 0.03 mmol) were dissolved in acetonitrile (3 mL). A 35 wt% aqueous hydrogen peroxide solution (206.3 μL, 2.4 mmol) was added to the reaction solution, and the mixture was reacted at 25 ° C. for 15 hours under an argon atmosphere. Thereafter, the crude product obtained by extraction with ether and drying under reduced pressure was subjected to medium pressure column chromatography (carrier: silica gel, hexane: EtOAc = 1: 0 → linear gradient of 2: 1 (v: v)). The product was separated and purified by This obtained compound 2 (12AC-3O) (72.7 mg, 52% yield).

(Production Examples 2 to 17)
The gem-dihydroperoxide compounds shown in Tables 1 to 3 were obtained in the same manner as in Production Example 1 except that the raw material ketones in Production Example 1 were changed to those shown in Tables 1 to 3. In Table 1-3, the numerical values of ^{"R 1"} and ^{"R 2"} are the respective number of carbon atoms of ^{"R 1"} and ^{"R 2"} in the formula (1).

(Structural analysis)
Each gem-dihydroperoxide compound prepared as described above was subjected to structural analysis by NMR (instrument name: AL400 spectrometer, ECA500 spectrometer instrument: manufactured by JEOL). In the table below, the melting point (mp) was determined by a micro melting point measuring device MP-S3 (manufactured by Anatech Yanaco).

<2. Antioxidant activity measurement (ESR)>
The gem-dihydroperoxide compound was treated with KF cells (5 × 10 ⁵ cells / ml) at 10 μM for 4 hours or 8 hours. Thereafter, the cells were washed with PBS, and the cells were collected by centrifugation. The recovered cells were completely lysed with 100 μL of 5-dimethyl-1-pyrroline-N-oxide (DMPO). The antioxidant activity of the prepared lysate was measured using JES-FA200 X-band spectrometer (manufactured by JEOL Ltd.). DMPO is used as a spin trap agent.

  The ESR spectra for dimethyl sulfoxide (DMSO), compound 2 (12AC-3O), and comparative compound 4 (11AC-3O) as a solvent for the gem-dihydroperoxide compound are shown in FIG.

  When the compound according to the present invention was used, the peak disappeared in the ESR spectrum unlike DMSO and Comparative Examples. Therefore, it can be seen that the compound according to the present invention has high antioxidant activity.

<3. Anticancer activity and cytotoxicity>
3-1. Culture of K562 cells RPMI1640 medium (manufactured by Wako Pure Chemical Industries, Ltd.) supplemented with 10% (v / v) heat-inactivated fetal calf serum (manufactured by Sigma) of human leukemia cell line K562 cells (purchased from ATCC) Incubation was performed under conditions of 5% carbon dioxide and 37 ° C. For human normal lymphocytes, lymphocytes were isolated by specific gravity centrifugation using Ficoll-Paque (manufactured by Invitrogen) and used for the test.

3-2. Counting the number of viable cells The cells prepared as described above were seeded in a multi-well plate so as to be 0.8 to 1.0 × 10 ⁵ cells / well. Three days after seeding, in order to induce cell proliferation, the cells were stimulated with 75 μM concanavalin A (Con-A) for 72 hours, and then the medium was changed. Thereafter, a gem-dihydroperoxide compound dissolved in DMSO was added to the medium so that the final concentration was 10 μM.

  After culturing the cells in a medium containing a gem-dihydroperoxide compound for 24 hours, the number of viable cells was counted. The number of viable cells was calculated by trypan blue dye exclusion test. Taking the number of viable cells in the group treated with DMSO as 100%, the ratio of viable cells in other test groups to the DMSO treated group was calculated (n = 6). The results are shown in FIG.

As shown in FIG. 2, it can be seen that the group treated with the compound according to the present invention has a small proportion of cancer cells. In the above formula (1), R ¹ is an alkyl group having 1 to ² carbon atoms; R ² is a saturated hydrocarbon group having 9 to 11 carbon atoms, and the total number of carbon atoms of R ¹ and R ² is 11 It can be seen that the compounds of ˜12 have particularly high anticancer activity. Compound 13 showed no cell survival.

  Furthermore, the number of living K562 cells treated with 10 μM compound 2 (12AC-3O) as described above was evaluated over time. FIG. 3 shows the result of calculating the number of viable cells with 100% before treatment.

  As shown in FIG. 3, it can be seen that the ratio of viable cells rapidly decreases after treatment with the compound according to the present invention.

  After treatment with 10 μM compound 2 (12AC-3O) for 48 hours as described above, the cells were observed with an electron microscope (product name H-7650, manufactured by Hitachi High-Technologies Corporation). The result is shown in FIG.

  As shown in FIG. 4, it can be seen that the cells treated with the compound according to the present invention have fragmented nuclei (arrows in FIG. 4).

3-3. Evaluation of apoptosis In order to analyze the mechanism of anticancer activity of gem-dihydroperoxide compounds, the presence or absence of apoptosis induction on cancer cells was morphologically evaluated. That is, K562 cells treated with 10 μM compound 2 (12AC-3O) for 4 or 8 hours as described above were stained with Hoechst 33342 according to the manufacturer's protocol (37 ° C., 60 minutes, 5 μg / ml Hoechst 33342, stock Washed with phosphate buffered saline (PBS, manufactured by Takara Bio Inc.). Cells were resuspended with PBS, the suspension was dropped onto a slide glass, and examined by fluorescence microscopy using a microscope (Olympus). The results are shown in FIG.

  In the group treated with DMSO, no nuclear fragmentation or condensation was observed, but in the group treated with Compound 2 (12AC-3O), nuclear fragmentation or condensation (white arrows in FIG. 5) was observed.

  Further, FIG. 6 shows the results of counting the number of cells undergoing apoptosis and calculating the ratio of cells undergoing apoptosis relative to the total number of cells.

  From the above, it was shown that apoptosis of cancer cells was induced in a short time after treatment by the compound according to the present invention.

3-4. Analysis of apoptosis-related protein Apoptosis-related protein was analyzed by Western blot using cells treated with 10 μM of compound 2 (12AC-3O) for 2, 4, 8, 12, or 24 hours.

(Western blot)
Ice-cold lysis buffer (10 mM Tris-HCl (pH 7.4), 1% (w / v) NP-40, 0.1% (w / v) deoxycholic acid, 0.1% (w / v) SDS , 150 mM NaCl, 1 mM EDTA, and 1% (w / v) protease inhibitor cocktail (Sigma-Aldrich)) and allowed to stand on ice for 20 minutes. The homogenate was centrifuged at 13,000 rpm for 20 minutes (4 ° C.), and the supernatant was collected as a whole cell protein sample. The protein content in the sample was measured using a DC protein assay kit (manufactured by Bio-Rad).

  Samples (10 μg protein amount) were separated by SDS-PAGE using 10.0 or 12.5% (w / v) polyacrylamide gel and transferred to a PVDF membrane (Perkin Elmer Life Sciences). Non-specific binding was blocked by incubation for 1 hour in 5% (w / v) skim milk (prepared in PBS containing 0.1% (w / v) Tween® 20 (PBS-T)). . Thereafter, an anti-PARP antibody (manufactured by Cell Signaling) or anti-caspase 8 appropriately diluted with PBS-T containing 2% (w / v) bovine serum albumin and 0.01% (w / v) sodium azide Membranes were incubated overnight at 4 ° C. with antibodies (Cell Signaling). The membrane was then washed 3 times with PBS-T and further incubated at room temperature (25 ° C.) with a secondary antibody (HRP-conjugated goat anti-rabbit antibody or HRP-conjugated horse anti-mouse IgG antibody, above Cell Signaling Technology). did. The membrane was then washed 3 times with PBS-T. Immunoblots were visualized using Amersham ECL plus Western blotting detection reagent (GE Healthcare). Β-actin was used as an internal standard by reincubating the same membrane with anti-β-actin antibody (Sigma-Aldrich). The result of the Western blot is shown in FIG.

  As shown in FIG. 7, it was found that caspase 8 was activated by the gem-dihydroperoxide compound according to the present invention, and the cleavage of PARP (poly (ADP-ribose) polymerase) proceeded. Therefore, the progress of apoptosis was also confirmed in the protein level test.

3-5. Cytotoxicity to normal cells Human normal lymphocytes were cultured under the same conditions as for K562 cells, stimulated with concanavalin A, and then treated with 5 μM or 10 μM compound 2 (12AC-3O) for 8 hours. Thereafter, the ratio of the living cells of the compound 2 (12AC-3O) treatment group to the DMSO treatment group was calculated as described above. The results are shown in FIG.

  In normal cells, the ratio of viable cells in the compound 2 (12AC-3O) treatment group was comparable to that in the DMSO treatment group. From the above, it can be seen that the compound according to the present invention has low cytotoxicity against normal cells.

Claims (8)

A compound represented by the following formula (1) or a physiologically acceptable salt thereof;
However, in the above formula (1), R ¹ is a substituted or unsubstituted, a straight-chain, branched-chain or a saturated hydrocarbon group having 1 to 4 carbon atoms containing a cycloaliphatic structure; R ² is a substituted or unsubstituted the linear, branched, or a saturated hydrocarbon group having 7 to 13 carbon atoms containing an alicyclic structure; R ¹ and Ri total 11-14 der number of carbon atoms of R ^2; wherein the substituted carbon The substituents in the saturated hydrocarbon group having 1 to 4 and the saturated hydrocarbon group having 7 to 13 carbon atoms are selected from the group consisting of a halogen atom, a hydroxyl group, an acyl group having 2 to 3 carbon atoms, and an alkoxy group having 1 to 3 carbon atoms. Ru is selected. The compound according to claim 1 or a physiologically acceptable salt thereof, wherein in the formula (1), R ¹ is an alkyl group having 1 to 2 carbon atoms. In the above formula (1), R ² is a substituted or unsubstituted, saturated hydrocarbon group having 9 to 11 carbon atoms including a linear, branched or alicyclic structure, and the number of carbon atoms of R ¹ and R ² Or a physiologically acceptable salt thereof according to claim 2, wherein the sum of is 11-12.   The antioxidant containing one or more selected from the compound of any one of Claims 1-3, and its physiologically acceptable salt as an active ingredient.   The anticancer agent which contains one or more selected from the compound of any one of Claims 1-3, and its physiologically acceptable salt as an active ingredient.   A cosmetic comprising one or more selected from the compounds according to any one of claims 1 to 3 and physiologically acceptable salts thereof.   A prophylactic and / or therapeutic agent for oxidative stress diseases, comprising as an active ingredient one or more selected from the compound according to any one of claims 1 to 3 and a physiologically acceptable salt thereof.   The prophylactic and / or therapeutic agent according to claim 7, wherein the oxidative stress disease is selected from the group consisting of neurodegenerative diseases, vascular disorders, heart diseases, inflammatory diseases, eye diseases, and skin diseases.