WO2011025270A2 - Metformin caffeic acid salt, method for preparing same, pharmaceutical composition comprising same, and combined formulation comprising same - Google Patents

Abstract

The present invention relates to a metformin caffeic acid salt, to a method for preparing same, to a pharmaceutical composition comprising same, and to a combined formulation comprising same. The metformin caffeic acid salt of the present invention has superior stability, enables easy tabletting, and has low toxicity, thus can be valuably used in the treatment of glycosuria, diabetes, metabolic syndrome, diabetes complications, menstrual irregularities, hypertension, hyperlipidemia, fatty liver, coronary artery disease, osteoporosis, polycystic ovary syndrome, cancer, myalgia, symptoms of muscle cell cytotoxicity, rhabdomyolysis, climacterium, etc., and/or provides antioxidant activity.

Description

Metformin caffeate, its preparation method, pharmaceutical composition comprising the same and a combination containing the same

The present invention relates to a novel salt of metformin, a method for preparing the same, a pharmaceutical composition comprising the same, and a combination formulation comprising the same.

N, N-dimethyl imidodicarbonimidic diamide (N, N-dimethyl imidodicarbonimidic diamide), commonly known as metformin, is an insulin-independent diabetes treatment drug. Is a biguanide-based drug that has the best hypoglycemic action and is most effective in preventing complications and worsening of symptoms.

Of all oral diabetes therapies, metformin is the only drug of choice. In particular, the finding that metformin's efficacy activates AMP-activated protein kinase (AMPK) has justified its clinical effectiveness. AMPK is a key enzyme that physiologically regulates carbohydrate and lipid metabolism. Metformin activates this enzyme to normalize hyperglycemia, improve lipid status, normalize menstrual irregularities, ovulation and pregnancy, and at the same time treat fatty liver and gene p53 (Tumor). protein p53) has been reported to prevent and treat cancer. The Cancer Cancer Journal of the Pennsylvania School of Medicine reported that metformin, an AMPK enzyme activator, was effective in the prevention and treatment of cancers lacking the p53 gene [Monica Buzzai, et al. Syntemic Treatment with the Antidiabetic Drug Metformin Selectively Impairs p53-Deficient Tumor Cell Growth, Cancer Res 2007; 67: (14). July 15, 2007].

In addition, metformin is a drug that activates AMPK enzymes to normalize sugar and lipid metabolism, and metformin administration in cancers lacking the p53 gene alters the energy metabolic pathways of cancer cells and increases anticancer activity in proportion to the dose of metformin. Metformin has been shown to be effective in treating cancer at normal doses for treating diabetes.

In addition, researchers from the Beth Israel Deaconess Medical center in Boston, USA, reported in a medical journal that metformin, a PGC-1α activator, prevents serious side effects such as myalgia, muscle cell damage, and even rhabdomyolysis. Effectiveness has been found [Jun-ichi Hanai, et al. The muscle-specific ubiquitin ligase atroigin-1 / MAFbx mediates statin-induced muscle toxicity, J. Clin. Invest. 117: 3940-3951 (2007). While expression of the Atrogin-1 gene causes myalgia, muscle cell damage, and rhabdomyolysis, metformin inhibits the expression of the Atrogin-1 gene due to PGC-1α transcription factor activity. To prevent and prevent muscle damage caused by.

In addition, Josie MM Evans published a study showing that the incidence of cancer in patients with type 2 diabetes is lower than those who do not receive metformin [Josie MM, Evans et al. BMJ. 2005 , 330, 1304-1305], Samantha L. Bowker reported that patients with type 2 diabetes who take metformin have a lower mortality associated with cancer than patients who take sulfonylureas or take insulin [Samantha L et al. Diabetes Care. 2006, 29, 254-258.

In addition, metformin has been reported to improve cardiovascular disease, lower triglycerides, low-density lipoprotein, and cholesterol levels in diabetics due to its antioxidant effects [UKPDS group. Effect of intensive blood-glucose control with metformin on complications in overweight Patients with type 2 diabetes (UKPDS 34) Lancet 1998: 352: 854-865].

Pharmaceutically, metformin is useful in the free base form, but it is administered in the form of a pharmaceutically acceptable acid addition salt because of the disadvantage of poor stability.

Accordingly, metformin is sold in the form of hydrochloride, and other metformin salts are metformin folate in Chinese Patent Publication No. CN 1962661A, metformin 1,2,6,7,8,8a-hexahydro- in WO 2005/033067. Beta, gamma, 6-trihydroxy-2-methyl-8-[(2s) -2-methyl-1-oxobutoxy]-, (betaR, gammaR, 1S, 2S, 6S, 8S, 8aS) -1-naphthaleneheptarate, metformin acetylsalicylate in US Pat. No. 3,957,853, metformin clofibrate salt in US Pat. No. 4,080,472, fumarate, succinate salt of metformin in US Pat. No. 6,031,004, p-chloro-phenoxy in Pat. Acetate salts, adamantane salts, and the like are disclosed in US Pat. No. 3,903,141.

However, the role of the "salt" in these metformin hydrochloride and other addition salts only had the effect of improving the physical properties of the pharmaceutical form, and does not increase the original pharmacological effect of metformin.

On the other hand, unlike conventional medicines, caffeic acid is a safe in vivo material with little reported toxicity.It is contained in medicinal plants such as fruits, such as pears containing coffee beans, basil, thyme, verbena, tarragon, oregano, and dandelion. Is a phenolic compound. Caffeic acid acts as an antioxidant and is known to be more effective than other antioxidants.

One technical problem to be solved by the present invention is to provide a metformin novel salt excellent in pharmacological effect.

Another technical problem to be solved by the present invention is to provide a method for producing a metformin novel salt.

Another technical problem to be solved by the present invention is diabetes, diabetes, metabolic syndrome, diabetes complications, menstrual irregularities, hypertension, hyperlipidemia, fatty liver, coronary artery disease, osteoporosis, polycystic ovary syndrome To provide a pharmaceutical composition for preventing, alleviating or treating a disease or condition selected from cancer, myalgia, myococytosis, rhabdomyolysis, or menopausal syndrome.

Another technical problem to be solved by the present invention is to provide an antioxidant pharmaceutical composition comprising a metformin novel salt as an active ingredient.

Another technical problem to be solved by the present invention is to provide a combination formulation comprising a metformin novel salt and a second drug.

The present invention provides metformin caffeate.

In the present invention, the binding molar ratio of metformin and caffeic acid salt is preferably 2: 1 to 1: 2, and more preferably metformin caffeic acid salt in a 1: 1 molar ratio of the following Chemical Formula 1.

[Formula 1]

In the present invention, metformin caffeate is meant to include all anhydrides, hydrates (hemihydrates, monohydrates, dihydrates, trihydrates, etc.), and solvates of metformin caffeate. The metformin caffeate may also be in crystalline form.

In addition, the present invention comprises the steps of 1) reacting the metformin hydrochloride with an inorganic base to obtain a metformin free base; And 2) reacting the metformin free base obtained in step 1 with the caffeic acid to produce metformin caffeate.

In the preparation method of the present invention, metformin hydrochloride used as starting material can be purchased commercially or prepared by a known method.

In the preparation method of the present invention, the inorganic base of step 1 means a conventional inorganic base used in the field of organic chemical manufacturing, for example, sodium hydroxide, potassium hydroxide, calcium hydroxide, lithium hydroxide, potassium carbonate, sodium carbonate, cesium carbonate, Sodium hydrogen carbonate, or potassium hydrogen carbonate, but examples thereof are not limited to the inorganic base that can be used in the present invention.

In the preparation method of the present invention, the use equivalent weight of the inorganic base is preferably 0.7 to 4 molar equivalents, more preferably 0.9 to 4 molar equivalents, and even more preferably 0.9 to 1.5 molar equivalents, relative to 1 molar equivalent of metformin hydrochloride. .

In step 1 of the preparation method of the present invention, the reaction temperature is preferably 0 to 60 ° C, more preferably 15 to 50 ° C. The reaction time is preferably 30 minutes to 3 hours, but may vary depending on the reaction temperature.

In step 2 of the preparation method of the present invention, caffeic acid may be purchased and used commercially.

In the preparation method step 2 of the present invention, the use equivalent of caffeic acid is preferably 0.7 to 4 molar equivalents, more preferably 0.9 to 4 molar equivalents, more than 0.9 to 1.5 molar equivalents, relative to 1 molar equivalent of metformin free base. desirable.

In step 2 of the preparation method of the present invention, the reaction temperature is preferably 0 to 80 ℃, the reaction temperature may vary depending on the type of reaction solvent. For example, when the reaction solvent is water, it is preferable to react at room temperature (15 to 30 ℃).

In the preparation method of the present invention, the reaction solvent is water or methanol, ethanol, n-propanol, isopropanol, n-butanol, t-butanol, pentanol, acetone, dioxane, dimethylformamide, dimethyl sulfoxide, acetonitrile, tetra Conventional organic solvents such as hydrofuran are used, with isopropanol being preferred in steps 1 and 2.

Specifically, metformin caffeic acid salt represented by Chemical Formula 1 may be prepared by the steps represented by Schemes 1 and 2.

Scheme 1

Scheme 2

In addition, the present invention is diabetes, diabetes, metabolic syndrome, diabetes complications, menstrual disorders, hypertension, hyperlipidemia, fatty liver, coronary artery disease, osteoporosis, polycystic ovary syndrome, cancer, myalgia, muscle cells containing metformin caffeate as an active ingredient Provided are pharmaceutical compositions for the prevention, alleviation or treatment of one or more diseases or symptoms selected from toxicosis and rhabdomyolysis, or menopausal syndrome. The prevention, alleviation or treatment may be due to the activating action of AMPKα. Preferably, it may be for simultaneously preventing, alleviating or treating cancer and diabetes.

The cancer may be a cancer lacking the p53 gene. In addition, the cancer may be cancer selected from the group consisting of uterine cancer, breast cancer, stomach cancer, brain cancer, rectal cancer, colon cancer, colon cancer, lung cancer, skin cancer, blood cancer, liver cancer, pancreatic cancer, prostate cancer and thyroid cancer, preferably colon cancer , Breast cancer or lung cancer.

In the present invention, diabetes includes not only diabetes but also diabetes of those with metabolic syndrome, which means that diabetes, obesity, hypertension, hyperlipidemia, fatty liver, coronary artery disease, osteoporosis or polycystic ovary syndrome are combined. . In addition, the complications of diabetes are the same as the disease or condition referred to as metabolic syndrome.

Preferably the composition may be for the prevention, alleviation or treatment of cancer. The cancer may be one or more selected from colon cancer, lung cancer or breast cancer.

The composition of the present invention may be for co-administration with a second drug. The second drug means another pharmaceutically effective ingredient other than metformin caffeic acid salt of the present invention.

Metformin caffeic acid salt of the present invention can be used for the treatment of various diseases as described above. Thus, the metformin caffeate of the present invention can be used in combination with a second drug for more efficient treatment of each disease. For example, the second drug may be an anticancer agent, an antihyperglycemic agent, an antiobesity agent, or the like. The antihyperglycemic agent may be for prevention, alleviation or treatment of diabetes mellitus, diabetes, metabolic syndrome, and / or diabetes complications due to hyperglycemia, and the antiobesity agent may be for prevention, alleviation or treatment of obesity. The antihyperglycemic agent may be at least one drug selected from the group consisting of biguanide-based drugs, sulfonylurea-based drugs, thiazolidione-based drugs, and alpha-glocosidase inhibitors. The anticancer agent is not limited as long as it can be used in combination with metformin caffeate, but may be a biological agent such as a gene therapy agent in addition to known chemotherapeutic agents such as alkylating agents, metabolic antagonists, natural agents, hormones, and / or antagonists, and / or immunotherapy agents. You can be the best. For example, nitrogen mustard, imatinib, oxaliplatin, rituximab, erlotinib, neratinib, lapatinib, zefitinib, vandetanib, nirotinib, semasanib, conservinib, axitinib, cediranib, Restautinib, trastuzumab, gefitinib, bortezomib, sunitinib, carboplatin, sorafenib, bevacizumab, cisplatin, cetuximab, biscumalboom, asparaginase, tretinoin, hydroxycarbamide , Dasatinib, estramastine, gemtuzumab ozogamycin, ibritumomab tucetan, heptaplatin, methylaminolevulinic acid, amsacrine, alemtuzumab, procarbazine, alprostadil, holmium nitrate chitosan , Gemcitabine, doxyfluridin, pemetrexed, tegapur, capecitabine, gimerasin, otheracil, azacytidine, methotrexate, uracil, cytarabine, fluorouracil, fludagabine, enositabine , Flutamide, decitabine, mer Captopurine, Thioguanine, Cladribine, Carmoper, Raltitrexed, Docetaxel, Paclitaxel, Irinotecan, Velotecan, Topotecan, Vinorelbine, Etoposide, Vincristine, Vinblastine, Teniposide, Doxorubicin, Ida Rubicin, epirubicin, mitoxantrone, mitomycin, bleomycin, daunorubicin, dactinomycin, pyrarubicin, aclarubicin, pepromycin, temozolomide, busulfan, ifosfamide, Cyclophosphamide, melfaran, altremin, dacarbazine, cheotepa, nimustine, chlorambucil, mitolactol, leucovorin, tretonin, xmestan, aminoglutesimid, anagrelide, Navelvin, padrazole, tamoxifen, toremifene, testosterone, anastrozole, letrozole, borosol, bicalutamide, romustine and / or carmustine and the like.

The antihyperglycemic agent is not limited as long as it can be used in combination with metformin ascorbate, but is not limited to acarbose, miglitol, bogliose, pramlinted, buformin, metformin, phenformin, allogliptin, sasagliptin, cytagliptin , Bildagliptin, Exenatide, Reroglutide, Insulin, Mytiglinide, Nateglinide, Repaglinide, Setohexamide, Carbutamide, Chlorpropamide, Glibenclamide, Glyborneuride, Glyclagit, glymepiride, glyphigit, glyquidone, glycentide, glysolamide, glycyclamide, tol azamide, tolbutamide, pioglitazone, rosiglitazone, and / or troglitazone.

The anti-obesity agent is not limited if it can be used in combination with metformin ascorbate, but not limited to dexamphetamine, caffeine, dexfenfluramine, diethylpropion, ephedrine, fenfluramine, fluoxetine, leptin, liraglutide, magdol, metformin, methylcellulose, oligo Start, penmetrazine, phentermine, phenylpropanolamine, limonabant, sibutramine, stuculia, sucrose, polyester, tefenfencin, topiramate, and / or zonamide.

The present invention also provides an antioxidant pharmaceutical composition containing metformin caffeate as an active ingredient. The antioxidant may be due to the activating action of AMPKα. The antioxidant may prevent, alleviate and / or treat cancer, aging, heart disease, brain disease, arteriosclerosis, inflammation, and / or immune diseases.

As used herein, the term "prevention" means any action that inhibits or delays the onset of a disease or condition by administration of a composition of the present invention. In the present invention, "treatment" means any action that improves or beneficially changes the symptoms of the disease by administration of the composition of the present invention, and "relaxation" means that the disease or symptom is no longer worsened by administration of the composition of the present invention. It means all actions.

The pharmaceutical compositions of the present invention may be in the form of conventional formulations that are acceptable and commonly used in the medical arts and may be, for example, tablets, soft capsules, hard capsules, pills, granules, powders, injections or solutions. The composition may be selected from sustained or immediate release. The sustained release means that the release of the active ingredient occurs slowly, including the base used for the purpose of sustained release in addition to the active ingredient, the immediate release means that the active ingredient is released immediately. Specifically, the tablet may be a sustained release tablet or an immediate release tablet.

In addition, the pharmaceutical compositions of the present invention may further comprise a pharmaceutically acceptable carrier, diluent or excipient.

Among the pharmaceutically acceptable carriers, a component selected from an enteric polymer, a hydrophobic substance, a hydrophilic polymer, and the like may be used as the matrix base used for sustained release. Examples of the enteric polymer include polyvinylacetate phthalate, hydroxypropylmethylcellulose phthalate, shellac, cellulose acetate phthalate, cellulose propionate phthalate, poly (methacrylic acid and methyl methacrylate) copolymer, and poly (methacrylic acid, ethylacrylic). Rate) copolymers and the like can be used one or more selected, preferably hydroxypropyl methyl cellulose phthalate is used.

The hydrophobic materials are pharmaceutically acceptable polyvinyl acetate, poly (methacrylate, methyl methacrylate) copolymer, poly (ethylacrylate, methyl methacrylate, trimethylaminoethylmethacrylate as polymethacrylate copolymers). Acrylates) copolymers, ethyl cellulose and cellulose acetate, fatty acids and fatty acid esters, fatty alcohols, waxes and inorganic substances, and the like, specifically, glyceryl palmitostearate, Fatty acid alcohols such as glyceryl stearate, glyceryl bihenate, cetyl palmitate, glyceryl monooleate and stearic acid, such as cetostearyl alcohol, cetyl alcohol and stearyl alcohol, as waxes, carnauba wax, beeswax Talc, sedimentation as an inorganic material, such as microcrystalline wax Selecting an acid, calcium hydrogen phosphate, calcium oxide, zinc oxide, titanium oxide, kaolin, bentonite, montmorillonite, and one or two or more selected from such bigeom and the like can be used.

The hydrophilic polymer may be selected from sugars, cellulose derivatives, gums, proteins, polyvinyl derivatives, polymethacrylate copolymers, polyethylene derivatives and carboxyvinyl polymers, and specifically, as the sugars, dextrin, polydextrin, dextran, Pectin and pectin derivatives, alginates, polygalacturonic acid, xylan, arabinoxylan, arabinogalactan, starch, hydroxypropyl starch, amylose, amylopectin and the like can be selected and used as cellulose derivatives. , Hydroxypropyl cellulose, hydroxymethyl cellulose, hydroxyethyl cellulose, methyl cellulose, carboxymethyl cellulose sodium, hydroxypropyl methyl cellulose acetate succinate, hydroxyethyl methyl cellulose and the like can be selected and used as a gum , Locker Soybean gum, tragacanta, carrageenan, acacia gum, gum arabic, gellan gum, xanthan gum, etc. can be selected and used, and gelatin, casein, zein, etc. can be selected as proteins, polyvinyl derivatives as polyvinyl derivatives Alcohol, polyvinyl pyrrolidone, polyvinyl acetal diethylamino acetate, and the like can be selected and used as the polymethacrylate copolymer, and poly (butyl methacrylate, (2-dimethylaminoethyl) methacrylate, methyl methacrylate Rate) copolymer, etc. can be selected and used, polyethylene glycol, polyethylene oxide, etc. can be selected and used as a polyethylene derivative, and a carbomer can be used as a carboxyvinyl polymer.

Starch, microcrystalline cellulose, lactose, glucose, mannitol, alginate, alkaline earth metal salt, clay, polyethylene glycol and / or dicalcium phosphate may be used as a pharmaceutically acceptable diluent without impairing the effects of the present invention. have.

As a binder, starch, microcrystalline cellulose, highly dispersible silica, mannitol, lactose, polyethylene glycol, polyvinylpyrrolidone, hydroxypropyl methylcellulose, hydroxypropylcellulose, natural gum, synthetic gum, copovidone and / or gelatin, etc. Can be used.

Starch or modified starch such as sodium starch glycolate, corn starch, potato starch or pregelatinized starch as a disintegrating agent, clays such as bentonite, montmorillonite, veegum, microcrystalline cellulose, hydroxypropyl cellulose or carboxy Celluloses such as methyl cellulose, algins such as sodium alginate or alginic acid, cross-linked celluloses such as croscarmellose sodium, gums such as guar gum and xanthan gum, and crosslinks such as crospovidone A polymer and boiling agents, such as sodium bicarbonate and a citric acid, can be selected and used. Lubricants include talc, magnesium stearate and alkaline earth metal stearate type calcium, zinc, lauryl sulfate, hydrogenated vegetable oils, sodium benzoate, sodium stearyl fumarate, glyceryl monostearate, polyethylene glycol 4000 and / or polyethylene glycol 6000 and the like can be used.

The scope of the present invention is not limited to the use of such additives, and the above additives may contain a conventional range of dosages by the choice of those skilled in the art.

As described above, metformin caffeate can be utilized as a preparation for oral administration in various forms. In the present invention, the dosage is preferably 50 to 3,000 mg metformin caffeate (based on metformin free base), but may vary depending on the age, sex, weight, nationality, health condition and degree of disease of the patient. Depending on the judgment, divided doses can also be administered once or twice a day.

In addition, the present invention is metformin caffeic acid diabetes, diabetes mellitus, metabolic syndrome, diabetes complications, menstrual irregularities, hypertension, hyperlipidemia, fatty liver, coronary artery disease, osteoporosis, polycystic ovary syndrome, cancer, myalgia, myococytosis, rhabdomyolysis or The present invention also provides a method for preventing, symptomatic or treating one or more diseases selected from menopausal syndrome. The present invention also provides an effective amount of metformin caffeate in diabetes mellitus, diabetes, metabolic syndrome, diabetic complications, menstrual irregularities, hypertension, hyperlipidemia, fatty liver, coronary Administering to a mammal, including a human, in need of preventing, alleviating or treating one or more diseases selected from arterial disease, osteoporosis, polycystic ovary syndrome, cancer, myalgia, myococytosis, rhabdomyolysis or menopausal syndrome. How to prevent or alleviate or treat the disease To provide.

The present invention also provides an antioxidant method comprising the step of administering to a mammal, including a human, in need of an effective amount of metformin caffeate. By the antioxidant method, cancer, aging, heart disease, brain disease, arteriosclerosis, inflammation, and / or immune diseases can be prevented or treated. As used herein, the term "administration" means introducing the metformin caffeate and / or pharmaceutical composition of the invention to a patient in any suitable manner, wherein the route of administration of the metformin caffeate and / or pharmaceutical composition of the invention is Administration can be by any general route as long as it can reach the desired tissue. Oral administration, intravenous administration, intramuscular administration, subcutaneous administration, endothelial administration, intranasal administration, pulmonary administration, rectal administration, intraperitoneal administration, intradural administration, but is not limited thereto.

In addition, the present invention is selected from diabetes, diabetes mellitus, metabolic syndrome, diabetic complications, menstrual irregularities, hypertension, hyperlipidemia, fatty liver, coronary artery disease, osteoporosis, polycystic ovary syndrome, cancer, myalgia, muscle cell toxicity, rhabdomyolysis or menopausal syndrome Metformin caffeate is provided for the prevention, alleviation or treatment of one or more diseases or conditions.

The present invention also provides metformin caffeate for antioxidant.

The matters mentioned in the compositions of the invention apply equally to the uses, methods of the invention, unless contradictory.

The present invention also provides a pharmaceutical formulation comprising the metformin caffeic acid salt of the present invention and a second drug.

In the formulations of the present invention, the same applies as long as the points mentioned in the compositions of the present invention do not contradict.

For formulation, reference may be made to Remington's Pharmaceutical Science (Recent), Mack Publishing Company, Easton PA, and others.

The active ingredient in the composition or formulation of the present invention may be included in 10 to 95% by weight based on the total weight.

Metformin caffeate and the composition of the present invention is diabetes, diabetes, metabolic syndrome, diabetes complications, menstrual irregularities, hypertension, hyperlipidemia, fatty liver, coronary artery disease, osteoporosis, polycystic ovary syndrome, cancer, myalgia, myococytosis, rhabdomyolysis And / or are effective in the treatment or prevention of menopausal syndrome and the like and have an antioxidant effect. In addition, the metformin caffeate salt of the present invention provides a crystalline acid addition salt suitable for the manufacture of pharmaceutical formulations, and thus has low toxicity by using a caffeate salt having a relatively low toxicity compared to conventional metformin hydrochloride prepared using hydrochloric acid. In addition, it has the effect of increasing the pharmaceutical and physical advantages such as solubility, stability, non-hygroscopicity and ease of processing of tablet formulations.

In addition, the metformin caffeate can be easily produced by the production method of the present invention.

In addition, the co-formulation of the present invention is excellent in pharmacological effects and can promote the convenience of the patient's medication.

Hereinafter, the present invention will be described in more detail with reference to Examples and Experimental Examples. These Examples and Experimental Examples are only for illustrating the present invention more specifically, and the scope of the present invention is not limited by these Examples and Experimental Examples.

In addition, the reagents and solvents mentioned below were purchased from Aldrich, USA and Daejin, Korea. ^{The 1} H-NMR and ¹³ C-NMR data are measured by Innova 600 MHz FT-NMR from Verian, USA, and the melting point (mp) is No. 1 of Electrothermal, UK. 9201 Melting point measured.

Example 1 Preparation of Metformin Caffeic Acid

Step 1: Preparation of Metformin Free Base

16.6 g of metformin hydrochloride (Harman Finochem) and 6.0 g of potassium hydroxide were added to 50 mL of isopropanol, and stirred at 50 ° C. for 2 hours. The reaction solution was cooled to 25 degrees Celsius, filtered, washed with 20 mL of isopropanol, and washed once more with 20 mL of acetone. The filtrate was concentrated and dried in vacuo to give 12.8 g (yield yield: 98.5%) of a white solid metformin freebase. Melting Point 119.0 ~ 119.5 ℃

¹ H-NMR (600 MHz, D ₂ O) δ (ppm) 3.07 (s, 6H)

¹³ C-NMR (150 MHz, D ₂ O) δ (ppm) 37.37, 158.56, 161.05

Step 2: Preparation of Metformin Caffeic Acid

To 5.0 g (1.2 equivalents) of metformin free base prepared in Step 1 of Example 1, 25 ml of isopropanol was added and dissolved. While stirring the reaction solution, 1.5 g (1.0 equivalent) of caffeic acid was slowly added dropwise, and then 100 mL of ethyl acetate was added thereto. After stirring for 2 hours at room temperature, the resulting crystals were filtered and washed with 20 mL of isopropanol and 50 mL of acetone. The filtrate was hot-air dried to obtain 4.8 g (yield yield: 61.7%) of metformin caffeate as a white solid. It was melting point 147.1 degreeC.

¹ H-NMR (600 MHz, D ₂ O) δ (ppm) 7.05 (d, J = 16.2 Hz, CH, 1H), 6.91 (d, J = 1.82 Hz, CH, 1H), 6.83 (dd, J = 2.4 Hz, J = 7.2 Hz, CH, 1H), 6.69 (d, J = 8.4 Hz, CH, 1H), 6.12 (d, J = 16.2 Hz, CH, 1H), 2.78 (s, CH ₃ , 6H)

¹³ C-NMR (150 MHz, D ₂ O) δ (ppm) 167.09, 160.07, 158.53, 145.85, 144.28, 140.88, 128.22, 121.74, 121.64, 116.30, 114.59, 37.49

Metformin caffeate is a crystalline acid addition salt suitable for the manufacture of pharmaceutical formulations, so it is not only toxic but also has low solubility, stability, and non-toxicity due to the use of caffeate, which is relatively less toxic than conventional metformin hydrochloride prepared using hydrochloric acid. It has the effect of increasing the pharmaceutical and physical advantages such as hygroscopicity and ease of processing of tablet formulations.

Example 2 Preparation of Metformin Caffeate-Containing Tablet

466.9 g of metformin caffeate prepared by the method of Example 1 and 143.1 g of microcrystalline cellulose were sieved through a No. 20 sieve, and then mixed in a V-type mixer for 60 minutes. Separately, 25 g of copovidone (Collidon VA64, BASF, Germany) and 10 g of hard silicic anhydride were sieved through a No. 35 sieve, added to the mixture, and mixed for 60 minutes. Finally, 5 g of stearic acid was sieved through a No. 35 sieve, added to the mixture, and mixed for 3 minutes.

Then, the final mixture was compressed into tablets containing 466.9 mg of metformin caffeate in one tablet.Opadry O3B28796 (hydroxypropylmethylcellulose 62.50%, oxidized by a high coater (SFC-30N, Sejong Machinery, Korea) Titanium 31.25%, polyethylene glycol 400 6.25%; Colorcon; USA) as a coating base to form a 20 mg film coating layer per tablet to prepare a tablet containing metformin caffeate.

Example 3 Preparation of Metformin Caffeate Containing Tablet

933.83 g of metformin caffeate and 36.17 g of mannitol, which were prepared by the method of Example 1, were sieved through a No. 20 sieve and then mixed in a high speed mixer for 3 minutes. Separately, 20 g of povidone was added to 120 g of isopropanol and dissolved to prepare a binding solution. The binding solution was administered to a high speed mixer, and then associated for 3 minutes. The associated union was dried in a steam dryer and sieved to No. 20 sieve to prepare granules. 5 g of hard silicic anhydride was sieved through a No. 35 sieve, added to the granules, and mixed in a V-type mixer for 60 minutes. Finally, 5 g of stearic acid was sieved through a No. 35 sieve, added to the mixture, and mixed for 3 minutes.

Subsequently, the final mixture was compressed into tablets containing 933.83 mg of metformin caffeate in one tablet, and a 20 mg film per tablet was used as a coating agent based on Opadry O3B28796 as a coater (SFC-30N, Sejong Machinery, Korea). A coating layer was formed to prepare metformin tablets containing metformin caffeate.

Example 4 Preparation of Metformin Caffeic Acid-Containing Sustained-Release Tablet

466.9 g of metformin caffeic acid, 88.1 g of dicalcium phosphate, and 400 g of hydroxypropylmethylcellulose 2208 (viscosity 200,000 cps) prepared in the same manner as in Example 1 were sieved through a No. 20 sieve and mixed for 60 minutes in a double cone mixer. The mixture was roller compacted under a pressure condition of 10-30 Mpa to prepare a slug, and was formed into a No. 20 sieve. Separately, 30 g of hydroxypropyl cellulose and 10 g of hard silicic anhydride were sieved through a No. 35 sieve, and added to the above sieved material, and mixed for 60 minutes. Finally, 5 g of magnesium stearate was sieved through a No. 35 sieve, added to the mixture, and mixed for 3 minutes.

Subsequently, the final mixture was compressed to prepare a sustained-release tablet containing 466.9 mg of metformin caffeine salt in one tablet, and a high coater (SFC-30N, Sejong Machinery, Korea) was coated with Opadry O3B28796 as a coating base. A coating layer was formed to prepare a metformin sustained-release tablet containing metformin taurine salt.

Example 5 Preparation of Metformin Caffeic Acid-Containing Sustained-Release Tablet

466.9 g of metformin caffeate, 48.1 g of microcrystalline cellulose and 440 g of polyethylene oxide (molecular weight 5 million, Dow Chemical, USA) were sieved through a No. 20 sieve and mixed in a double cone mixer for 45 minutes. . Separately, 30 g of copovidone (Collidon VA 64, BASF, Germany) and 10 g of hard silicic anhydride were sieved through a No. 35 sieve, added to the mixture, and mixed for 45 minutes. Finally, 5 g of magnesium stearate was sieved through a No. 35 sieve, added to the mixture, and mixed for 3 minutes.

Subsequently, the final mixture was compressed to prepare a sustained-release tablet containing 466.9 mg of metformin caffeate in one tablet. As a high coater (SFC-30N, Sejong Machinery, Korea), Opadry O3B28796 was used as a coating base, and 20 mg A film coating layer was formed to prepare metformin tablets containing metformin taurine salt.

Example 6 Preparation of Metformin Caffeate-Containing Capsule

767.72 g of metformin caffeate and 224.28 g of microcrystalline cellulose, respectively, were sieved through a No. 20 sieve and mixed in a V-type mixer for 60 minutes. 4 g of hard silicic anhydride was sieved through a No. 35 sieve, added to the mixture, and mixed for 60 minutes. Finally, 4 g of stearic acid was sieved through a No. 35 sieve, added to the mixture, and mixed for 3 minutes.

The final mixture was then filled into capsules to prepare capsules containing 191.93 mg of metformin caffeate in one capsule.

Example 7 Preparation of Metformin Caffeic Acid Salt and Capecitabine-Containing Co-Formulations

466.9 g of metformin caffeate, 150 g of capecitabine and 153.1 g of microcrystalline cellulose, respectively, were sieved through a No. 20 sieve and mixed in a V-type mixer for 60 minutes. Separately, 25 g of copovidone (collidone VA6, BASF, Germany) and 5 g of hard silicic anhydride were sieved through a No. 35 sieve, added to the mixture, and mixed for 60 minutes. Finally, 5 g of stearic acid was sieved through a No. 35 sieve and added to the mixture and mixed for 3 minutes.

Subsequently, the final mixture was compressed into tablets containing 466.9 mg of metformin caffeate and 150 mg of capecitabine in one tablet, and was coated with Opadry O3B28796 as a high coater (SFC-30F, Sejong Machinery, Korea). 20 mg of the film coating layer was formed to prepare a film-coated tablet containing metformin caffeate and capecitabine.

Experimental Example

Experimental Example 1: Confirmation of cancer cell proliferation inhibitory effect of metformin caffeate

Metformin caffeate synthesized in the manner described in Example 1 of the present invention was treated with cancer cells to determine the effect of inhibiting cancer cell proliferation. The experimental method is as follows.

Using MCF7 cell line derived from human breast cancer, HCT116 cell line derived from human colon cancer, and A549 cell line derived from human lung cancer, MTT (3- (4,5-dimethylthiazole-2-yl) -2,5- By ditetrazolium bromide assay, the survival rate (%) of the cells was measured to determine the effect of metformin caffeate on cancer cell proliferation.

96 wells of MCF7 cell line (Korea Cell Line Bank), HCT116 cell line (Korea Cell Line Bank), A549 cell line (Korea Cell Line Bank) in Dulbecco's modified Eagle's medium (DMEM) medium containing 10% calf serum Incubated for about 24 hours in a plate (well plate). Thereafter, metformin caffeate synthesized in the manner described in Example 1 was treated with 0.4 mM for MCF7 and HCT116 cells, 2 mM for A549 cells, and cultured for 72 hours. In addition, in order to obtain GIC50, metformin caffeate 10mM, 2mM, 0.4mM, 0.08mM, and 0.016mM were treated in the same cultures for three cell lines and incubated for 72 hours.

After the metformin caffeate treatment, MTT was added to the culture medium to identify living cells and further cultured for 3 hours. The resulting formazan crystal was dissolved using dimethyl sulfoxide (DMSO), and then the absorbance of the solution was measured at 560 nm.

After 72 hours of incubation, the number of cells surviving in the well plate treated with metformin caffeate compared to the number of cells cultured in the well plate not treated with metformin caffeate was expressed as cell viability (%). In addition, the concentration of metformin caffeate (GIC 50) in which growth was inhibited to 50% using a cell viability curve was calculated to confirm the effect of inhibiting cancer cell proliferation of metformin caffeate, and the results are shown in Tables 1 and 2, respectively. Indicated.

In addition, the cell survival rate (%) and GIC 50 value was obtained using metformin hydrochloride or caffeic acid instead of metformin caffeate, and the results are shown in Table 1 and Table 2, respectively.

Table 1 Cell survival rate (%)

Table 2 Cell Growth Inhibition Concentration (GIC 50)

As can be seen from the cell viability of Table 1, when HCT116, MCF7 and A549 cells were cultured in the presence of metformin caffeate, the cell survival rate was only 18-48% compared to the case of culture under metformin hydrochloride. Able to know.

These results indicate that metformin caffeate inhibits various cancer cells 2 to 5.6 times more effectively than metformin hydrochloride. It was found that metformin caffeate inhibited the survival of cancer cells, particularly breast cancer, colon cancer, and lung cancer cells, more effectively than metformin hydrochloride.

As can be seen from the GIC 50 values of Table 2, metformin hydrochloride was able to inhibit 50% growth of HCT116, MCF7, and A549 cells when metformin caffeate was used at 0.27 mM, 0.21 mM, and 0.6 mM, respectively. It can be seen that the treatment of 4.5mM, 10mM or 3.5mM in the same cell, respectively, can inhibit the growth of the cell by 50%.

From this, it was found that metformin caffeate could inhibit the growth of cancer cells derived from breast cancer, colon cancer and lung cancer more effectively than metformin hydrochloride using metformin hydrochloride.

The present invention provides metformin caffeic acid salt, a method for preparing the same, a pharmaceutical composition comprising the same, and a combination formulation containing the same. Metformin caffeate of the present invention has excellent stability and ease of processing of tablets, low toxicity, diabetes, diabetes, metabolic syndrome, diabetes complications, menstrual irregularities, hypertension, hyperlipidemia, fatty liver, coronary artery disease, osteoporosis, polycystic ovary syndrome It is effective in cancer, myalgia, muscle cytotoxicity, rhabdomyolysis, menopausal syndrome, and / or antioxidant, and has industrial applicability.

Claims (19)

Metformin Caffeic Acid. According to claim 1, Metformin caffeic acid salt of the formula (1). [Formula 1]

3. Metformin caffeic acid according to claim 2, wherein the melting point is 147.1 degrees Celsius. Diabetes, diabetes mellitus, metabolic syndrome, diabetes complications, menstrual irregularities, hypertension, hyperlipidemia, fatty liver, coronary artery disease, osteoporosis, polycystic ovary syndrome, comprising the metformin caffeate of any one of claims 1 to 3, A pharmaceutical composition for the prevention, alleviation or treatment of one or more diseases or symptoms selected from cancer, myalgia, myococytosis, rhabdomyolysis, or menopausal syndrome. The composition of claim 4, wherein the cancer is a cancer lacking the p53 gene. The composition of claim 4, wherein the cancer is a cancer selected from the group consisting of uterine cancer, breast cancer, stomach cancer, brain cancer, rectal cancer, colon cancer, colon cancer, lung cancer, skin cancer, blood cancer, liver cancer, pancreatic cancer, prostate cancer and thyroid cancer. The composition of claim 4 for use in combination with a second drug. 8. The composition of claim 7, wherein the second drug is at least one selected from anticancer agents, antihyperglycemic agents, or antiobesity agents. The method of claim 8, wherein the anticancer agent is nitrogen mustard, imatinib, oxaliplatin, rituximab, erlotinib, neratinib, lapatinib, zefitinib, vandetanib, nirotinib, semasanib, conservinib, axity Nip, cediranib, restautinib, trastuzumab, gefitinib, bortezomib, sunitinib, carboplatin, sorafenib, bevacizumab, cisplatin, cetuximab, biscumalboom, asparaginase, tretinoin , Hydroxycarbamide, dasatinib, estramastin, gemtuzumab ozogamycin, ibritumab tucetan, heptaplatin, methylaminolevulinic acid, amsacrine, alemtuzumab, procarbazine, alprosta Dill, Holmium Nitrate Chitosan, Gemcitabine, Doxyfluidine, Pemetrexed, Tegapur, Capecitabine, Gimerasine, Oteraceyl, Azacytidine, Methotrexate, Uracil, Citarabine, Fluorouracil, Fludagar Vienna, Enositabine, Flu Mead, decitabine, mercaptopurine, thioguanine, cladribine, carmofer, raltitrexed, docetaxel, paclitaxel, irinotecan, velotecan, topotecan, vinorelbine, etoposide, vincristine, vinblastine, teni Fosid, doxorubicin, idarubicin, epirubicin, mitoxantrone, mitomycin, bleomycin, daunorubicin, dactinomycin, pyrarubicin, aclarubicin, pepromycin, temozolomide, laying Pan, Ifosfamide, Cyclophosphamide, Melparan, Altretmin, Dcarbazine, Chiotepa, Nimustine, Chlorambucil, Mitolactol, Leucovorin, Tretonin, Xmestan, Aminoglutesimid At least one selected from the group consisting of anagrelide, nabelbin, padrazole, tamoxifen, toremifene, testosterone, anastrozole, letrozole, borosol, bicalutamide, romustine and carmustine . The pharmaceutical composition according to claim 4, wherein the pharmaceutical composition is prepared from tablets, soft capsules, hard capsules, pills, granules, powders, injections or solutions. The composition of claim 10, wherein the composition is selected from sustained or immediate release. The pharmaceutical composition according to claim 4, wherein the content of metformin caffeate is 50 to 3,000 mg. The composition of claim 4 for preventing, alleviating or treating cancer. The composition of claim 13, wherein the cancer is one or more selected from colon cancer, lung cancer, or breast cancer. The pharmaceutical composition for antioxidant containing the metformin caffeate of any one of claims 1 to 3 as an active ingredient. The composition of claim 15, wherein the composition is for preventing, alleviating, or treating one or more diseases or symptoms selected from cancer, aging, heart disease, brain disease, arteriosclerosis, inflammation, or immune disease. A combination formulation comprising the metformin caffeate salt of any one of claims 1 to 3 and a second drug. 18. The formulation of claim 17, wherein the second drug is one or more selected from anticancer agents, antihyperglycemic agents, or antiobesity agents. 19. The method of claim 18, wherein the anticancer agent is nitrogen mustard, imatinib, oxaliplatin, rituximab, erlotinib, neratinib, lapatinib, zefitinib, vandetanib, nirotinib, semasanib, conservinib, axity Nip, cediranib, restautinib, trastuzumab, gefitinib, bortezomib, sunitinib, carboplatin, sorafenib, bevacizumab, cisplatin, cetuximab, biscumalboom, asparaginase, tretinoin , Hydroxycarbamide, dasatinib, estramastin, gemtuzumab ozogamycin, ibritumab tucetan, heptaplatin, methylaminolevulinic acid, amsacrine, alemtuzumab, procarbazine, alprosta Dill, Holmium Nitrate Chitosan, Gemcitabine, Doxyfluidine, Pemetrexed, Tegapur, Capecitabine, Gimerasine, Oteraceyl, Azacytidine, Methotrexate, Uracil, Citarabine, Fluorouracil, Fludagar Vienna, Enositabine, Flu Tamide, decitabine, mercaptopurine, thioguanine, cladribine, carmofer, raltitrexed, docetaxel, paclitaxel, irinotecan, velotecan, topotecan, vinorelbine, etoposide, vincristine, vinblastine, Teniposide, doxorubicin, idarubicin, epirubicin, mitoxantrone, mitomycin, bleomycin, daunorubicin, dactinomycin, pyrarubicin, aclarubicin, pepromycin, temozolomide, Busulfan, ifosfamide, cyclophosphamide, melfaran, altretmin, dacarbazine, cheotepa, nimustine, chlorambucil, mitolactol, leukovorin, tretonin, exemstan, aminogluteci One or more selected from the group consisting of mead, anagrelide, nabelbin, padrazole, tamoxifen, toremifene, testosterone, anastrozole, letrozole, borosol, bicalutamide, romustine and carmustine Formulation.