KR20110021693A - Metformin ascorbic acid salt, preparation thereof, pharmaceutical composition comprising the same and combined formulation comprising the same - Google Patents

Abstract

PURPOSE: A metformin ascorbate and a pharmaceutical composition containing the same are provided to ensure excellent stability and proceesibility of tablet. CONSTITUTION: A metformin ascorbate is denoted by chemical formula 1. The melting point of metformin ascorbate is 145 degree celsius. A pharmaceutical composition for preventing or treating diabetes, metabolic syndrome, diabetic complication, hypertension, fatty liver, or cancer contains metformin ascorbate of chemical formula 1 as an active ingredient. The composition is combined with a second drug, such as anti-cancer drug, anti-hypertension agent, or anti-obesity agent. The pharmaceutical composition is manufactured in the form of tablet, soft capsule, hard capsule, pill, granule, powder, injection, or liquid. The composition is sustained release or rapid release.

Description

Metformin ascorbate, a method for preparing the same, a pharmaceutical composition comprising the same, and a combination comprising the same {Metformin ascorbic acid salt, preparation conjugate, 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, which 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 metabolism and lipid metabolism. Metformin activates this enzyme to normalize hyperglycemia, improve lipid status, normalize menstrual irregularities, ovulation and pregnancy, and 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 normalizes sugar and lipid metabolism by activating the AMPK enzyme. When metformin is administered to cancers lacking the p53 gene, the energy metabolic pathways of cancer cells are changed and anticancer activity increases 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 rate 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.

In addition, ascorbic acid is known as vitamin C as one of the water-soluble vitamins, and the development and sale of health drinks and functional foods is continuously expanding as a safe substance with little reported toxicity.

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 ascorbate.

In the present invention, the binding molar ratio of metformin and ascorbate is preferably from 2: 1 to 1: 2, and more preferably from 1: 1 molar ratio of metformin ascorbate of formula (1).

[Formula 1]

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

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, ascorbic acid may be purchased and used commercially.

In step 2 of the preparation method of the present invention, the ascorbic acid use equivalent 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 ℃), and in the case of isopropanol, it is preferred to react by heating to 50 to 60 ℃.

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, methanol is preferred in step 1 and water or isopropanol is preferred in step 2.

Specifically, metformin ascorbate 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 is diabetes, diabetes, metabolic syndrome, diabetes complications, dysmenorrhea, hypertension, hyperlipidemia, fatty liver, coronary artery disease, osteoporosis, polycystic ovary syndrome, muscle cell toxicity, muscle cell toxicity Provided is a pharmaceutical composition for the prevention, alleviation or treatment of one or more diseases or symptoms selected from symptom 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 lung 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 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 ascorbate of the present invention. The metformin ascorbate of the present invention can be used for the treatment of various diseases as described above. Thus, the metformin ascorbate 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 ascorbate, 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. Can be. 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, Carmofer, 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 ascorbate 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 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 and 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 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 Etc. 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 ascorbate can be utilized as a preparation for oral administration in various forms. In the present invention, the dosage is preferably from 50 to 3,000 mg of metformin ascorbate (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 may be given once or twice a day.

In addition, the present invention is metformin ascorbate diabetes mellitus, diabetes, metabolic syndrome, diabetic complications, menstrual irregularities, hypertension, hyperlipidemia, fatty liver, coronary artery disease, osteoporosis, polycystic ovary syndrome, cancer, myalgia, myococytosis, rhabdomyolysis or Provided are one or more disease prophylactic, symptomatic or therapeutic uses selected from menopausal syndrome.

In addition, the present invention is metformin ascorbate 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, rhabdomyolysis Or administering to a mammal comprising a human in need of preventing, alleviating or treating one or more diseases selected from menopausal syndrome.

The present invention also provides an antioxidant method comprising administering to a mammal, including a human, in need of an effective amount of metformin ascorbate.

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 ascorbate and / or pharmaceutical composition of the invention to a patient in any suitable manner, wherein the route of administration of the metformin ascorbate and / or pharmaceutical composition of the invention is intended for Administration can be by any general route as long as it can be reached. 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 ascorbate is provided for the prevention, alleviation or treatment of one or more diseases or conditions.

The present invention also provides metformin ascorbate 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 ascorbate 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.

The metformin ascorbate and the composition of the present invention exhibit excellent AMPKα activating action, diabetes, diabetes, metabolic syndrome, diabetes complications, menstrual irregularities, hypertension, hyperlipidemia, fatty liver, coronary artery disease, osteoporosis, polycystic ovary syndrome, cancer, myalgia It is effective in the treatment or prophylaxis of muscle cell toxicity, rhabdomyolysis and / or menopausal syndrome and has an antioxidant effect. In addition, the metformin ascorbate of the present invention provides a crystalline acid addition salt suitable for the preparation of pharmaceutical formulations, which is low toxicity by using ascorbate having a relatively low toxicity compared to conventional metformin hydrochloride prepared using hydrochloric acid. 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, metformin ascorbate 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.

Figure 1 shows the IR spectrum for metformin ascorbate in accordance with one embodiment of the present invention.
2 is a graph confirming the activation effect of AMPKα of metformin ascorbate 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. Measured by the 9201 melting point meter, the FT IR is measured by the TRAVEL-IR model manufactured by SENSIR Technologies of the United States.

< Example  1> Metformin Ascorbate  Produce

Step 1: Metformin Free base  Produce

200.00 g of metformin hydrochloride (Harman Finochem) and 48.30 g of sodium hydroxide were added to 1 L of methanol, and the resulting inorganic salt was filtered by stirring at room temperature for 2 hours, and the filtrate was concentrated under reduced pressure. 2 L of acetone was added to the resulting solid, and the insoluble matters were not dissolved. The filtrate was concentrated under reduced pressure to obtain 148.9 g of a metformin free base (yield yield: 95.5%). 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 Ascorbate  Produce

3.38 g of the metformin free base obtained in Step 1 of Example 1 was added to 50 mL of isopropanol and warmed to 60 degrees Celsius while stirring. After confirming that all the metformin free bases were dissolved, 4.44 g of ascorbic acid was added and stirred for 1 hour at 60 degrees Celsius. After the reaction solution was dissolved, it was confirmed that a white solid was produced. 50 mL of acetone was added to the reaction solution, cooled to 5 degrees Celsius, and the resulting solid was filtered. The resulting crystals were washed with 5.0 mL of isopropanol and 5.0 mL of acetone, followed by hot air drying at 50 ° C. for 12 hours to obtain 3.87 g of yellow solid metformin ascorbate (yield yield: 50.3%). The melting point was 145.0 ° C., and its IR data is shown in FIG. 1.

¹ H-NMR (600MHz, D ₂ O) δ (ppm) 2.77 (s, 6H), 3.47 (m, 2H), 3.75 (m, 1H), 4.23 (d, 2H)

¹³ C-NMR (150 MHz, D ₂ O) δ (ppm) 37.47, 62.57, 69.59, 78.40, 113.32, 158.51, 160.08, 175.65, 177.67

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

< Example  2> metformin Ascorbate  Produce

0.77 g of the metformin free base obtained in Step 1 of Example 1 and 0.88 g of ascorbic acid were added to 3.5 mL of purified water at room temperature, followed by stirring for 1 hour. After confirming that all of the reaction solution was changed to clear, it was lyophilized for 15 hours using a freeze dryer.

The resulting solid was crushed finely using a mortar and then 5 mL of acetone was added. Ultrasonic wave was added for about 10 minutes, and the resulting solid was washed with 50 mL acetone and 50 mL isopropanol and hot-air dried at 50 ° C. for 12 hours to obtain 1.09 g of yellow solid metformin ascorbate (yield yield: 71.4%). The melting point was 145.0 ° C., and the values of ¹ H-NMR and ¹³ C-NMR were the same as in Example 1.

< Example  3> metformin Ascorbate  Preparation of Containing Tablets

460.8 g of metformin ascorbate prepared by the method of Example 1 and 209.2 g of microcrystalline cellulose were respectively sieved through a No. 20 sieve and mixed in a V-type mixer for 60 minutes.

Separately, 20 g of copovidone (collidone 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.

Subsequently, the final mixture was compressed into tablets containing 380.8 mg of metformin ascorbate in one tablet.Opadry O3B28796 (hydroxypropylmethylcellulose 62.50%, titanium oxide) using a high coater (SFC-30N, Sejong Machinery, Korea) 31.25%, polyethylene glycol 400 6.25%; Colorcon; USA) as a coating base to form a film coating layer of 15mg per tablet to prepare a tablet containing metformin ascorbate.

< Example  4> Metformin Ascorbate  Preparation of Containing Tablets

921.63 g of metformin ascorbate prepared by the method of Example 1 was sieved through a No. 35 sieve and then charged into a high speed mixer. Separately, 35 g of povidone was added to 120 g of isopropanol and dissolved to prepare a binding solution. The binding solution was fed 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. 8.37 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.

The final mixture was then compressed into tablets containing 921.63 mg of metformin ascorbate in one tablet. Was formed to prepare a metformin tablet containing metformin ascorbate.

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

A double cone mixer after sieving 230.4 g of metformin ascorbate, 94.6 g of microcrystalline cellulose, and 250 g of hydroxypropylmethylcellulose 2208 (viscosity 100,000 cps, Shin-etsu, Japan), respectively, prepared in the same manner as in Example 1 Mix for 60 minutes at. Separately, 15 g of hydroxypropyl cellulose 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, added to the mixture, and mixed for 3 minutes.

Subsequently, the final mixture was compressed to prepare a sustained-release tablet containing 380.8 mg of metformin ascorbate in one tablet. The coating layer was formed to produce a sustained release metformin tablet containing metformin ascorbate.

< Example  6> Metformin Ascorbate  contain Sustained release  Manufacture of tablets

460.8 g of metformin ascorbate, 59.2 g of dicalcium phosphate, and 445 g of hydroxypropylmethylcellulose 2208 (viscosity 100,000 cps) 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 condition of 10-30 Mpa to prepare a slug, and was formed into a No. 20 sieve. Separately, 20 g of copovidone (collidone VA64, BASF, Germany) and 10 g of hard silicic anhydride were sieved through a No. 35 sieve, and added to the above formulation, 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 380.8 mg of metformin ascorbate in one tablet, and a high coater (SFC-30N, Sejong Machinery, Korea) was coated with Opadry O3B28796 as a coating base. The coating layer was formed to produce a sustained release metformin tablet containing metformin ascorbate.

< Example  7> Metformin Ascorbate  contain Sustained release  Manufacture of tablets

460.8 g of metformin ascorbate, 64.2 g of microcrystalline cellulose and 400 g of polyethylene oxide (5 million molecular weight, Dow Chemical, USA) prepared in the same manner as in Example 1 were sieved through a No. 20 sieve and then mixed in a double cone mixer for 60 minutes. . Separately, 40 g of hydroxypropyl cellulose and 10 g of hard silicic anhydride were sieved through a No. 35 sieve and 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.

Subsequently, the final mixture was compressed to prepare a sustained-release tablet containing 380.8 mg of metformin ascorbate in one tablet, and a high coater (SFC-30N, Sejong Machinery, Korea) was coated with Opadry O3B28796 as a coating base. The coating layer was formed to produce a sustained release metformin tablet containing metformin ascorbate.

< Example  8> Metformin Ascorbate  Preparation of containing capsule

921.6 g of metformin ascorbate and 84.4 g of microcrystalline cellulose prepared in the same manner as in Example 1 were sieved through a No. 20 sieve, and then mixed in a V-type mixer for 60 minutes. 8 g of hard silicic anhydride was sieved through a No. 35 sieve, added to the mixture, and mixed for 60 minutes. Finally, 6 g of stearic acid was sieved through a No. 35 sieve and added to the mixture, followed by mixing for 3 minutes.

Subsequently, the final mixture was filled into capsules to prepare capsules containing 230.4 mg of metformin ascorbate in one capsule.

< Example  9> Metformin Ascorbate and Capecitabine  Of containing complex preparations Produce

460.8 g of metformin ascorbate prepared in the same manner as in Example 1, 150 g of capecitabine, 100 g of mannitol, and 159.2 g of microcrystalline cellulose were sieved through a No. 35 sieve and then charged into a high speed mixer. Separately, 20 g of povidone was added to 120 g of isopropanol and dissolved to prepare a binding solution. 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 460.8 mg of metformin ascorbate and 150 mg of capecitabine in one tablet, and coated with Opadry O3B28796 as a high coater (SFC-30N, Sejong Machinery, Korea). A film coating layer of 15 mg per tablet was formed to prepare a film coated tablet containing metformin ascorbate and capecitabine.

[ Experimental Example ]

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

Metformin ascorbate 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 A549 cell line derived from human lung cancer, MTT (3- (4,5-dimethylthiazole-2-yl) -2,5-ditetrazoliumbromide) assay was used to determine the percentage of cell survival rate of metformin ascorbate. The cancer cell proliferation inhibitory effect was confirmed.

A549 cell line (Korea Cell Line Bank) was incubated for 24 hours in a 96 well plate so that each cell number was approximately 5000 in DMEM (Dulbecco's modified Eagle's medium) containing 10% calf serum. Thereafter, 2mM and 10mM of metformin ascorbate, synthesized in the manner described in Example 1, were treated in the culture solution and cultured for 72 hours. In addition, in order to obtain GIC 50, metformin ascorbate 10 mM, 2 mM, 0.4 mM, 0.08 mM, 0.016 mM were respectively treated in the culture solution and incubated for 72 hours.

MTT was added to the culture solution to identify the living cells after metformin ascorbate treatment 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 ascorbate compared to the number of cells cultured in the well plate not treated with metformin ascorbate was expressed as cell viability (%). In addition, the concentration of metformin ascorbate (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 ascorbate, and the results are shown in Table 1 and Table 2, respectively. It was.

In addition, metformin hydrochloride or ascorbic acid was used instead of metformin ascorbate to obtain cell viability (%) and GIC 50 values, and the results are shown in Tables 1 and 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 A549 cells were cultured in the presence of metformin ascorbate, the cell viability was lower than when cultured under metformin hydrochloride, from which metformin ascorbate was a cancer cell than metformin hydrochloride. In particular, it was found that the survival of lung cancer cells is effectively suppressed.

In addition, as can be seen from the GIC 50 value of Table 2, the use of metformin ascorbate could inhibit the growth of A549 cells by 50%, whereas metformin hydrochloride had to be treated with 3.5 mM to increase the cell growth by 50%. Could be restrained. Therefore, it was found that the use of metformin ascorbate in a smaller amount than metformin hydrochloride can effectively inhibit the growth of cancer cells, especially cancer cells derived from lung cancer.

Experimental Example  2: metformin Ascorbate AMPK Confirmation of the activation effect of α

The effect of activating AMPKα was measured by treating cells with metformin ascorbate 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 of metformin ascorbate was confirmed in MCF 7 cell line (Korea Cell Line Bank) derived from human breast cancer cells using AMPKα immunoassay kit (Invitrogen, catalog No. KHO0651). It was.

The MCF 7 cell line (Korea Cell Line Bank) was cultured in DMEM medium containing 10% calf serum, placed in a 6-well plate so that the number of cells was about 5 × 10 ⁵ , and then incubated with 5% CO _2. Cells were cultured. The culture solution was treated with 2 mM metformin ascorbate, and then cultured for 24 hours.

Phosphorylation of threonine 172 residue (T172) of AMPKα in cells cultured in the presence of metformin ascorbate and cells cultured without metformin ascorbate 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 threonine 172 residue of AMPKα from the cell lysate was used and the results are shown in Table 3 and FIG. 2. The ratio of the degree of phosphorylation of cells cultured in the presence of metformin ascorbate to the degree of phosphorylation of cells cultured in medium without metformin ascorbate is shown in Table 3.

[Table 3]

As can be seen in Table 3 and Figure 2, it can be seen that metformin ascorbate effectively activates AMPKα. Thus, metformin ascorbate may be used to treat diabetes, diabetes, metabolic syndrome, diabetic complications, menstrual irregularities, hypertension, hyperlipidemia, fatty liver, coronary artery disease, osteoporosis, polycystic ovary syndrome, cancer, myalgia, myococytosis, rhabdomyolysis, and It can be seen that it may have an excellent effect on diseases such as menopausal syndrome.

Claims (19)

Metformin ascorbate. 2. The metformin ascorbate of claim 1, wherein
[Formula 1]

The metformin ascorbate of claim 2, wherein the melting point is 145.0 degrees Celsius. Diabetes, diabetes mellitus, metabolic syndrome, diabetes complications, menstrual irregularities, hypertension, hyperlipidemia, fatty liver, coronary artery disease, osteoporosis, polycystic ovary syndrome, cancer containing the metformin ascorbate of any one of claims 1 to 3 as an active ingredient Pharmaceutical composition for the prevention, alleviation or treatment of one or more diseases or symptoms selected from 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 6, wherein the cancer is lung cancer The composition of claim 4 for use in combination with a second drug. The composition of claim 8, wherein the second drug is one or more selected from anticancer agents, antihyperglycemic agents, or anti-obesity agents. 10. The method of claim 9, 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 11, wherein the composition is selected from sustained or immediate release. The pharmaceutical composition according to claim 4, wherein the content of metformin ascorbate is 50 to 3,000 mg. The composition of claim 4 for preventing, alleviating or treating cancer. An antioxidant pharmaceutical composition containing the metformin ascorbate 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 a disease or condition selected from cancer, aging, heart disease, brain disease, arteriosclerosis, inflammation, or immune disease. A co-formulation comprising the metformin ascorbate of claim 1 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 At least one selected from the group consisting of mead, anagrelide, nabelbin, padrazole, tamoxifen, toremifene, testosterone, anastrozole, letrozole, borosol, bicalutamide, romustine and carmustine Formulation.

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