KR20110021691A - Metformin methanesulfonic acidsalt, preparation thereof, pharmaceutical composition comprising the same and combined formulation comprising the same - Google Patents

Abstract

PURPOSE: A metformin methanesulfonate and a pharmaceutical composition comprising the same are provided to ensure excellent activation of AMPKα, relatively low toxicity, stability, and non-hygroscopic property. CONSTITUTION: A metformin methanesulfonate represented by chemical formula 1 has a melting point of 194.0~194.5 °C. A pharmaceutical composition contains the metformin methanesulfonic acid salt as an active ingredient. The metformin methanesulfonate can be effectively used in treating 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 and/or climacterium, etc.

Description

Metformin methanesulfonic acidsalt, preparations, pharmaceutical composition comprising the same and combined formulation comprising 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) is commonly known as metformin. 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 alone has been characterized as the first choice in many papers. 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, is 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 patients with type 2 diabetes have a lower incidence of cancer 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 have metformin have lower cancer-related mortality than patients who take sulfonylureas or 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.

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 and menopausal syndrome.

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 methanesulfonate.

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

[Formula 1]

In the present invention, metformin methanesulfonate is meant to include all anhydrides, hydrates (hemihydrates, monohydrates, dihydrates, trihydrates, etc.), solvates, and the like of metformin methanesulfonate. The metformin methanesulfonate 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 methanesulfonic acid to produce metformin methanesulfonic acid salt.

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 usable 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, methanesulfonic acid may be purchased and used commercially.

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

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 acetone, 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 step 1 and acetone being preferred in step 2.

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

Scheme 1

[Formula 2] [Formula 3]

Scheme 2

[Formula 3] [Formula 1]

In addition, the present invention diabetes, diabetes, metabolic syndrome, complications of diabetes mellitus, dysmenorrhea, hypertension, hyperlipidemia, fatty liver, coronary artery disease, osteoporosis, polycystic ovary syndrome, muscle cells, muscle cells containing metformin methanesulfonate 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 Can be.

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 metabolic syndrome or diabetes.

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 methanesulfonate of the present invention. The metformin methanesulfonate of the present invention can be used for the treatment of various diseases as described above. Thus, the metformin methanesulfonate 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 methanesulfonic acid salt, 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, estramastin, gemtuzumab ozogamycin, ibritumomab tucetan, heptaplatin, methylaminolevulinic acid, amsacrine, alemtuzumab, procarbazine, alprostadil, holmium nitrate , Gemcitabine, doxyfluridin, pemetrexed, tegapur, capecitabine, gimerasin, otheracil, azacytidine, methotrexate, uracil, cytarabine, fluorouracil, fludagabine, enositabine , Flutamide, decitabine, mer Topurine, thioguanine, cladribine, carmoper, raltitrexed, docetaxel, paclitaxel, irinotecan, velotecan, topotecan, vinorelbine, etoposide, vincristine, vinblastine, teniposide, doxorubicin, 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.

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 exacerbated by the 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, hydroxypropylmethyl cellulose phthalate, shellac, cellulose acetate phthalate, cellulose propionate phthalate, poly (methacrylic acid, methyl methacrylate) copolymer, and poly (methacrylic acid, ethylacrylic). Rate) copolymers and the like can be used one or more, 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 and used as proteins, 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, poly (methacrylic acid, methyl methacrylate) copolymer, etc. can be selected and used, polyethylene glycol, polyethylene oxide, etc. can be selected and used as a polyethylene derivative, and carbomer can be used as a carboxyvinyl polymer. Can be.

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 methanesulfonate can be utilized as a preparation for oral administration in various forms. In the present invention, the dosage is preferably metformin methanesulfonate (based on metformin free base) 50 to 3,000 mg, 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 diabetes, diabetes, metabolic syndrome, complications of diabetes mellitus, dysmenorrhea, hypertension, hyperlipidemia, fatty liver, coronary artery disease, osteoporosis, polycystic ovary syndrome, muscle cells, muscle cells containing metformin methanesulfonate as an active ingredient Provided for the prevention, alleviation or treatment of one or more diseases or symptoms selected from toxicosis and rhabdomyolysis or menopausal syndrome.

In addition, the present invention is metformin methanesulfonate effective amount of diabetes, diabetes mellitus, metabolic syndrome, diabetic complications, menstrual disorders, hypertension, hyperlipidemia, fatty liver, coronary artery disease, osteoporosis, polycystic ovary syndrome, cancer, myalgia, muscle cell toxicity, rhabdomyomus Provided is a method for preventing, alleviating, or treating a disease comprising administering to a mammal, including a human, in need of prophylaxis, symptomatic relief, or treatment of at least one disease selected from lysis or menopausal syndrome.

As used herein, the term "administration" means introducing the metformin methanesulfonate or pharmaceutical composition of the present invention to a patient in any suitable manner, wherein the route of administration of the metformin methanesulfonate or pharmaceutical composition of the present invention reaches the target tissue. As far as possible, administration may be via conventional routes. 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, diabetes complications, menstrual irregularities, hypertension, hyperlipidemia, fatty liver, coronary artery disease, osteoporosis, polycystic ovary syndrome, cancer, myalgia, myococytosis, rhabdomyolysis or menopausal syndrome Metformin methanesulfonate is provided for the prevention, alleviation or treatment of one or more diseases or conditions.

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 metformin methanesulfonate 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 methanesulfonate and the composition of the present invention exhibits excellent AMPKα activation action, diabetes, diabetes, metabolic syndrome, diabetes complications, menstrual irregularities, hypertension, hyperlipidemia, fatty liver, coronary artery disease, osteoporosis, polycystic ovary syndrome, cancer, It is effective for the treatment or prevention of myalgia, myococytosis, rhabdomyolysis, and / or menopausal syndrome. In addition, the metformin methanesulfonic acid salt of the present invention provides a crystalline acid addition salt suitable for the preparation of pharmaceutical formulations, and thus has low toxicity using methanesulfonic acid salt, which is relatively less toxic than 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 methanesulfonic acid salt 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.

1 is a graph confirming the activation effect of AMPKα of metformin methanesulfonic acid salt according to an embodiment of the present invention.

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> Metformin Methanesulfonate  Produce

Step 1: Metformin Free base  Produce

16.6 g of metformin hydrochloride (Harman Finochem) and 6.0 g of sodium 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 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: Metformin Methanesulfonate  Produce

150 ml of acetone was added to 10.0 g (1 equivalent) of the metformin free base prepared in Step 1 of Example 1 to dissolve it. While stirring the reaction solution, 8.9 g (1.2 equivalents) of methanesulfonic acid (mesylic acid, Methanesulfonic acid) was slowly added dropwise and stirred at room temperature for 2 hours. 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 14.6 g (yield yield: 84%) of metformin methanesulfonate as a white solid. Melting point 194.0-194.5 degreeC.

¹ H-NMR (600 MHz, D ₂ O) δ (ppm) 2.82 (s, CH ₃ , 3H), 2.74 (s, CH ₃ , 6H)

¹³ C-NMR (150 MHz, D ₂ O) δ (ppm) 163.0, 40.1, 34.5

Metformin methanesulfonate 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 using methanesulfonate, 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> metformin Methanesulfonate  Preparation of Containing Tablets

340.03 g of metformin methanesulfonic acid salt prepared by the method of Example 1 and 93.97 g of microcrystalline cellulose were respectively sieved through a No. 20 sieve and mixed in a V-type mixer for 60 minutes.

Separately, 10 g of copovidone (collidone VA64, BASF, Germany) and 3 g of hard silicic anhydride were sieved through a No. 35 sieve, added to the mixture, and mixed for 60 minutes. Finally, 3 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 340.03 mg of metformin methanesulfonic acid salt in one tablet.Opadry O3B28796 (hydroxypropylmethylcellulose 62.50%, oxidized by a high coater (SFC-30N, Sejong Machinery, Korea) Tablets containing metformin methanesulfonate were prepared by forming a film coating layer of 10 mg per tablet based on 31.25% titanium, 6.25% polyethylene glycol 400; Colorcon; USA).

< Example  3> metformin Methanesulfonate  Preparation of Containing Tablets

680.08 g of metformin methanesulfonic acid salt and 62.92 g of dicalcium phosphate, respectively, 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, 25 g of povidone was added to 150 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. 7 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.

Then, the final mixture was compressed into tablets containing 680.08 mg of metformin methanesulfonic acid salt 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 methanesulfonate.

< Example  4> Metformin Methanesulfonate  contain Sustained release  Manufacture of tablets

340.03 g of metformin methanesulfonate prepared in the same manner as in Example 1, 49.97 g of microcrystalline cellulose, and 300 g of polyethylene oxide (5 million molecular weight, 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 (collidone VA 64, BASF, Germany) and 5 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 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 a sustained-release tablet containing 340.03 mg of metformin methanesulfonate in one tablet, and 15 mg per tablet was coated with Opadry O3B28796 as a coating agent (SFC-30N, Sejong Machinery, Korea). A film coating layer was formed to prepare metformin tablets containing metformin methanesulfonate.

< Example  5> Metformin Methanesulfonate  contain Sustained release  Manufacture of tablets

340.03 g of metformin methanesulfonate, 99.97 g of dicalcium phosphate, and 340 g of hydroxypropylmethylcellulose 2208 (viscosity 100,000 cps), which were prepared in the same manner as in Example 1, were sieved through a No. 20 sieve and mixed in a double cone mixer for 60 minutes. The mixture was roller compacted under a pressure of 15-25 Mpa to prepare a slug, and was formed into a No. 20 sieve. Separately, 25 g of hydroxypropyl cellulose and 10 g of hard silicic anhydride were sieved through a No. 35 sieve, added to the above-mentioned formulations, 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.

Then, the final mixture was compressed into a sustained-release tablet containing 340.03 mg of metformin methanesulfonic acid salt in one tablet, and 20 mg per tablet was coated with Opadry O3B28796 as a coating agent (SFC-30N, Sejong Machinery, Korea). A film coating layer was formed to prepare a sustained release metformin tablet containing metformin methanesulfonate.

< Example  6> Metformin Methanesulfonate  Preparation of containing capsule

340.04 g of metformin methanesulfonic acid salt and 97.96 g of microcrystalline cellulose were respectively sieved through a No. 20 sieve and mixed in a V-type mixer for 60 minutes. 2 g of hard silicic anhydride was sieved through a No. 35 sieve, added to the mixture, and mixed for 60 minutes. Finally, 2 g of magnesium stearate 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 170.02 mg of metformin methanesulfonate in one capsule.

< Example  7> Metformin Methanesulfonic acid salts Bicalutamide  Of containing complex preparations Produce

340.03 g of metformin methanesulfonate, 50 g of bicalutamide, and 93.97 g of microcrystalline cellulose, respectively, were sieved through a No. 20 sieve and mixed in a V-type mixer for 60 minutes. Separately, 10 g of copovidone (collidone VA64, BASF, Germany) and 3 g of hard silicic anhydride were sieved through a No. 35 sieve, added to the mixture, and mixed for 60 minutes. Finally, 3 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 340.03 mg of metformin methanesulfonate and 50 mg of bicalutamide in 1 tablet, and was coated with Opadry O3B28796 as a high coater (SFC-30F, Sejong Machinery, Korea). A film coating layer of 10 mg per tablet was formed to prepare a film-coated tablet containing metformin propionate and bicalutamide.

[ Experimental Example ]

Experimental Example  1: metformin Methanesulfonate AMPK Confirmation of the activation effect of α

The effect of activating AMPKα was measured by treating cells with metformin methanesulfonate synthesized in the manner described in Example 1 of the present invention. The experimental method is as follows.

AMPKα (5′-AMP-activated protein kinase alpha) activation effect of metformin methanesulfonate on MCF 7 cell line (Korea Cell Line Bank) derived from human breast cancer cells using AMPKα immunoassay kit (Invitrogen, catalog No. KHO0651) Confirmed.

The MCF 7 cell line (Korea Cell Line Bank) was cultured in Dulbecco's modified Eagle's medium (DMEM) medium containing 10% calf serum and placed in a 6-well plate with about 5 × 10 ⁵ cells. The cells were cultured in an incubator fed with CO ₂ . The culture was treated with 10 mM metformin methanesulfonate and then incubated for 24 hours.

Phosphorylation of threonine 172 residue (T172) of AMPKα in cells cultured in the presence of metformin methanesulfonate and cells cultured without metformin methanesulfonate was confirmed using an AMPKα immunoassay kit (Invitrogen, catalog No. KHO0651). After lysing the cells in the manner suggested by the AMPKα immunoassay kit (Invitrogen, catalog No. KHO0651), the method was described in the instructions for use of the AMPKα immunoassay kit. The degree of phosphorylation of the threonine 172 residue of AMPKα from the cell lysate was used and the results are shown in Table 1 and FIG. 1. The ratio of the degree of phosphorylation of cells cultured in the presence of metformin methanesulfonate to the degree of phosphorylation of cells cultured in medium without metformin methanesulfonate is shown in Table 1.

In addition, the degree of phosphorylation of threonine 172 residue (Thr172) of AMPK was confirmed using metformin hydrochloride instead of metformin methanesulfonate, and the results are shown in Table 1 below.

TABLE 1

As can be seen in Table 1 and FIG. 1, it was found that at 10 mM metformin methanesulfonate, phosphorylation of threonine 172 residues of AMPKα was greater than metformin hydrochloride. Therefore, it can be seen that metformin methanesulfonate activates AMPKα more effectively than metformin hydrochloride. Therefore, metformin methanesulfonate can be used for diabetes, diabetes, metabolic syndrome, diabetic complications, menstrual irregularities, hypertension, hyperlipidemia, fatty liver, coronary artery disease, osteoporosis, polycystic ovary syndrome, cancer, myalgia, myococytosis, rhabdomyolysis, And / or excellent effects on diseases such as menopausal syndrome.

Experimental Example  2: metformin Methanesulfonate  Confirmation of cancer cell proliferation inhibitory effect

Metformin methanesulfonate 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 HCT116 cell line derived from human colon cancer, MTT (3- (4,5-dimethylthiazole-2-yl) -2,5-ditetrazoliumbromide) assay measures the percent survival of cells and metformin methanesulfonate The cancer cell proliferation inhibitory effect was confirmed.

HCT116 cell line (Korea Cell Line Bank) was incubated for about 24 hours in a 96 well plate so that each cell number was approximately 5000 in DMEM medium containing 10% calf serum. Thereafter, 10 mM of metformin methanesulfonate synthesized in the manner described in Example 1 was treated to the culture solution and cultured for 72 hours to see cell viability.

After the metformin methanesulfonic acid 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 methanesulfonate compared to the number of cells cultured in the well plate not treated with metformin methanesulfonate was expressed as% cell viability. The results are shown in Table 2.

In addition, the cell survival rate (%) was calculated using metformin hydrochloride or methanesulfonic acid instead of metformin methanesulfonate, and the results are shown in Table 2.

Table 2 Cell survival rate (%)

As can be seen from the cell viability of Table 2, when HCT116 cells were cultured in the presence of metformin methanesulfonate, the cell survival rate was lower than when cultured under metformin hydrochloride, from which metformin methanesulfonate metformin hydrochloride It was found that more effectively inhibit the survival of cancer cells, particularly colon cancer cells.

Claims (16)

Metformin methanesulfonate. According to claim 1, Metformin methanesulfonic acid salt of formula (1).
[Formula 1]

3. Metformin methanesulfonate according to claim 2, wherein the melting point is 194.0 to 194.5 degrees Celsius. Diabetes, diabetes mellitus, metabolic syndrome, diabetic complications, menstrual irregularities, hypertension, hyperlipidemia, fatty liver, coronary artery disease, osteoporosis, polycystic ovary syndrome, comprising the metformin methanesulfonate of any one of claims 1 to 3 as an active ingredient, A pharmaceutical composition for preventing, alleviating or treating 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, gemcitabine, doxyfluidine, pemetrexed, tegapur, capecitabine, gimerasin, oteraceyl, azacytidine, methotrexate, uracil, cytarabine, 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 methanesulfonate is 50 to 3,000 mg. The composition of claim 4 for preventing, alleviating or treating cancer and diabetes. A combination formulation comprising the metformin methanesulfonate of any one of claims 1 to 3 and a second drug. The formulation of claim 14, wherein the second drug is at least one selected from anticancer agents, antihyperglycemic agents, or anti-obesity agents. The method of claim 15, 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 At least one selected from the group consisting of mead, anagrelide, nabelbin, padrazole, tamoxifen, toremifene, testosterone, anastrozole, letrozole, borosol, bicalutamide, romustine and carmustine Formulation.

Images