KR100516648B1 - A lignan compound and a hypoglycemic agent comprising the lignan compound as active ingredient - Google Patents

Abstract

The present invention is excellent in hypoglycemic activity prepared from isolated thistle ( Carduus crispus L.), a wide range of new compounds: (7R, 8R, 8'R, 9'R)-4, 7, 4 ', 9'-tetrahydroxy-3,3'-dimethoxy- ^{1,3,5,1 ', 3', 5 '} -8 . 8 ', 9 . O. 9'-Lignan and a novel antidiabetic agent containing the substance as an active ingredient.

      The new antidiabetic agent according to the present invention is expected to have an excellent hypoglycemic activity and a very high safety compared to the drugs widely used as an oral hypoglycemic agent, and also to significantly reduce side effects compared to the existing drugs.

Description

A lignan compound and a hypoglycemic agent comprising the lignan compound as active ingredient

The present invention relates to a novel lignanic compound exhibiting a hypoglycemic action first isolated from natural sources. In more detail, the present invention is isolated from the Carduus crispus L. outpost (especially flower buds), and then formula (7R, 8R, 8'R, 9'R) -4, 7 , 4 ', 9'-tetrahydroxy-3, 3'-dimethoxy- ^{1,3,5,1 ', 3', 5 '} -8 . 8 ', 9 . O. 9'-Lignan: (+)-Todolactol A (hereinafter SJ-2030) and a new oral hypoglycemic agent containing this substance as an active ingredient.

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The invention material SJ-2030 is a lignan-based material having a structure of dibenzylbutyrolactone , which was first isolated from Roger ( Taxus willichiana ) in 1982 by Isoliovil [ J. Nat. . prod . 45 (1), 78 (1982)], but this material could not determine the stereoconfiguration of C-7, C-8, C-8 'and C-9' in its structure. Subsequently, Ozawa et al., 1988, isolated from Abies sacchalinensis to 8R, 8'R, and 9'R in Isoliovil, todolactol A [ Mokuzai Gakkaishi 34 (10), 851 (1988)] Todolactol A was an epimer mixture of 7R and 7S in the stereoconfiguration of the hydroxyl group bound to C-7. In 1994, Barrero et al . Reported the separation of 7S-todolactol A tetraacetate body from the CHCl ₃ extract fraction of Abies pinsapo as a whole acetate body [ J. Nat. Prod. 57 (6), 713 (1994)]. However, the inventors have isolated SJ-2030, an inventive compound from the fin thistle ( Carduus crispus L.), which shows a cross-peak with H-8 'and H-8' and Hb-7 'in the NOESY spectrum. It can be seen that the H-7 of this compound is in the same plane as H-8 ', as shown in Fig. 1, and the [α] value is +31.68. R-type (7R, 8R, 8'R, 9'R)-4, 7, 4 ', 9'-tetrahydroxy-3, 3'-dimethoxy- ^{1,3,5,1 ', 3', 5 '} -8 . 8 ', 9 . O. 9'-Lignan, identified as (+)-todolactol A, is the first foreign substance to be separated from nature.

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(SJ-2030) (Todolactone A (SJ-2031)) Isorioville

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       Diabetes is a chronic metabolic disease that causes dysfunction of nerves, kidneys, and retinas, thereby degrading quality of life and causing one of life-threatening diseases. Most of the adult patients are classified as insulin independent diabetes mellitus. Patients use a combination of drugs and diet for the treatment of disease. However, the most popular oral hypoglycemic agent for the treatment of insulin independent diabetes is sulfonylurea system. And biguanide-based drugs, which have problems with their effectiveness and safety, and in particular have serious side effects such as hypoglycemia and lactic acidosis, as well as liver dysfunction. Therefore, to reduce these problems and side effects, improve the metabolic disorders, there is an urgent need for the development of high-safety diabetes treatment for long-term use.

     The present inventors conducted a drug efficacy search on natural resources to develop an anti-diabetic agent that can be ideally used in clinical practice. Among them, the extracts of the fin thistle flower group showed the strong hypoglycemic action, and the active ingredient was traced and separated by the fractionation and separation method according to the physiological activity orientation of this plant, and the lignan compound SJ-2030 has the hypoglycemic effect. It was confirmed that it is an excellent material.

The fin thistle ( Carduus crispus L.) is a biennial herb belonging to the genus Compositae, growing straight up to 60-100 cm, with wings on the main stem, with spiky serrates. The leaves are regenerated, long oval, erupted, deeply and shallowly divided into pterygium, and the lateral side is dull, ending with thorns. Flowers bloom in June-August, purple but white, and fruit is July-September. It is distributed throughout the country and grows in mountainous or wild areas. Their outposts or roots are called astringents or parenterals. It is also called cotton (wooden), fugu (비), unsecured lake, jatacho, cerebrum (채 公 菜). Big wind. Colloquial. It has the effect of anti-inflammatory, and is mainly a sense of wind fever, wind fever, wind cramps, skin ulcers, lobster, urethritis, and carbuncle (癰 腫), arthritis (關節炎), vaginal damage (跌打 損傷). It is widely used for burns, hematuria, and hemostasis.

      The present invention is described in detail as follows.

     Finned thistle flower parts were extracted with a mixed solvent of water and alcohol, the extract was concentrated under reduced pressure, and systematic solvent fractionation was performed. Each of the fractionated solutions was concentrated to prepare a concentrated extract, which was used as a primary drug search sample. SJ-2030 having a hypoglycemic effect was isolated by performing silica gel column chromatography and gel filtration on the methylene chloride fraction showing a significant effect. SJ-2030 showed excellent efficacy and high safety as a result of comparative experiments on the drugs used for oral hypoglycemic and the hypoglycemic effect and acute toxicity. It is expected. SJ-2030 is conventionally used in the form of tablets, capsules, powders, oral solutions, poultices, and injections, which are conventionally used pharmaceutical forms with excipients or auxiliaries, stabilizers, PH regulators, antioxidants, etc. It can be administered in the form of a formulation of. The amount of SJ-2030 may vary depending on the patient's condition, age, sex, etc., but in general, the amount of 10-2000 mg may be divided into several times a day. This component may also be administered alone or in combination with other effective agents in the form of commonly used acids, alkali salts or inorganic or organic acid salts.

      The present invention is explained in more detail by the following examples and experimental examples.

Example 1.

     Finned thistle fire department (10 kg) was cut into small pieces, reflux-cooled extracted with methanol (30 L) for 3 hours, and the extract was dried under reduced pressure to obtain a concentrated extract. Distilled water (2 L) was added to the extract and suspended in a water bath, followed by systematic solvent fractionation at room temperature, followed by nucleic acid (9 g), methylene chloride (15 g), ethyl acetate (23 g), and butanol (67 g). Five extracts, such as and water (93 g), were prepared and used as a sample for the primary drug search. In addition, methylene chloride fractions with significant activity were subjected to silica gel column chromatography using chloroform and methanol, gel filtration using methanol as a solvent, and recrystallized with water and methanol to isolate the blood glucose lowering substance SJ-2030. It was.

Example 2.

     5 kg of the fin thistles were chilled with 15 L of 80% methanol for 72 hours, filtered and concentrated under reduced pressure. The concentrated extract was suspended in water, fractionated with ether, and the remaining water layer was partitioned sequentially with ethyl acetate, butanol, and water to obtain respective extracts. The ether fraction, an active fraction, was subjected to silica gel column chromatography using benzene, chloroform and methanol, and gel filtration using methanol, and recrystallized from water and methanol to separate SJ-2030.

Example 3.

     The cut and cut thistle flower parts are extracted with 80% ethanol three times at room temperature for 24 hours, the extracts are combined and concentrated appropriately, the solution is partitioned between nucleic acid and 90% methanol, and the 90% methanol layer is concentrated and 60 The methylene chloride layer obtained by fractional extraction with% methanol and methylene chloride was concentrated under reduced pressure, and then the concentrated extract was subjected to silica gel column chromatography using benzene, chloroform and methanol, and gel filtration using methanol. Recrystallization with methanol separated SJ-2030.

Instrument Analysis Data of SJ-2030

Colorless crystal

* mp: 165-167 ℃

[Α] ²⁵ _d : +31.68 (c = 0.25, MeOH)

* UV λmax (MeOH) nm: 230, 282

       λmax (MeOH + NaOMe) nm: 250, 290

* IR νmax (KBr) cm-1: 3350, 1610, 1520, 1470, 1450, 1430, 1270, 1240 1160, 1120, 1090, 1040, 1010, 930, 850, 810 and 780

HR-MS m / z: 376.1528, Calcd. for C ₂ 0H ₂₄ O ₇ 376.1510

* EI-MS (70eV) m / z (rel. Int.,%):

         376 (M +, 15), 358 (20), 340 (11), 206 (33), 175 (21), 153 (62), 137 (100)

* ¹ H-NMR (500MHz, DMSO-d ₆ )

* ¹³ C-NMR (125MHz, DMSO-d ₆ )

* DEPT (125MHz, DMSO-d ₆ )

* C-H COSY (HMQC)

* Extensive C-H COSY (HMBC) ------------------ Table 1

<Figure 1>

NMR Instrumental Analysis Data of SJ-2030 No. C δc DEPT δ H, mult., J in HZ HMBC 123456789 135.85110.28147.29145.41114.88118.6074.1351.5467.48 CCHCCCHCHCHCHCH ₂ 6.83, s 6.71, d, 8.06.69, dd, 7.1, 1.04.47, dd, 7.4, 4.42.16, m3.53, dd, 13.3, 5.4 C-1, C-3, C-4, C-6, C-7 C-1, C-3, C-4C-1, C-2, C-4C-2, C-6, C-9, C-1C-7, C-8'C-9 ' 1'2'3'4'5'6'7'a 131.34112.95147.27144.54115.18120.9538.73 CCHCCCHCHCH ₂ 6.70, d, 1.2 6.65, d, 8.16.53, dd, 7.7, 1.82.59, dd, 13.6, 4.5 C-4 ', C-6' C-1 ', C-3'C-2', C-4'C-1 ', C-2', C-6 ', C-8', C-9 ' 7'b8'9'3-OCH ₃ 50.29101.5555.5155.48 CHCHCHCH ₃ 2.39, dd, 12.0, 10.62.32, m4.95, d, 4.73.73, s C-1 ', C-2', C-6 ', C-8', C-9'C-1'C-8, C-9C-3 3'-OCH ₃ 4-OH4'-OH7-OH9'-OH CH ₃ 3.74, s8.79, s8.67, s5.40, d, 4.55.89, d, 5.1 C-3'C-3, C-4, C-5C-3 ', C-4', C-5'C-7, C-8C-8 ', C-9'

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Experimental Example 1

     Acute Toxicity Experiment

Abnormal behavioral behavior by oral administration of component SJ-2030 in a group of 10 pure ICR male males (23 ± 2 g) fasted from the evening before the experiment. The presence or absence and LD ₅₀ values were calculated according to the methods of Richfield J. and Wilcoxon F. from the mortality rate of 72 hours.

Acute Toxicity Experimental group LD ₅₀ (mg / kg) Route of administration     SJ-2030     Tolbutamide *        > 1000> 500 280 185        Oral cavity oral cavity

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* Where to get it (Drug Pharmaceutical Information Center, Japan, 10/1997).

From the results of Table 2, the LD ₅₀ value of SJ-2030 was more than 1000 mg / kg for oral administration and 500 mg / kg for intraperitoneal administration, which is very safe compared to tolbutamide.

Experimental Example 2

      Hypoglycemic action (hyperglycemic-induced mice)

     The male rats of 25 ± 2 g of body weight were divided into 10 groups and fasted for 16 hours, followed by intravenous administration of 75 mg / kg of alloxane, 48-72 hours later, and the blood glucose level was 400 mg / dl or more. The rats were selected and used for experiments. SJ-2030 was dissolved in physiological saline and administered orally, and blood glucose was measured by glucose oxidase method after collecting blood from the tail vein after 120 minutes.

Blood sugar measurement Experimental group Capacity (mg / kg, po ) Marisu Glucose content in serum (mg / dl) 0 hours 2 hours  %     Relative control group (aloxane) SJ-2031SJ-2030       --40204080    101010101010    126.4 ± 4.0 521.8 ± 35.2 520.9 ± 31.0 510.4 ± 33.3 516.8 ± 35.3 530.8 ± 26.7     127.4 ± 5.0 498.8 ± 52.6 405.4 ± 38.2 * 437.8 ± 50.6 * 345.5 ± 43.6 ** 310.6 ± 44.0 **     0.8 4.4 22.2 14.2 33.1 41.5

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* p <0.05, ** p <0.01

      From the results of Table 3, it was confirmed that the hypoglycemic action of SJ-2030 in the hyperglycemic mice was strongly dose dependent.

Formulation Example 1

* Manufacture of tablets

   SJ-2030 50 mg

   Cornstarch 30 mg

   Lactose 30 mg

   Sodium starch glycolate 30 mg

   H, P, C 5 mg

   Magnesium Stearate 5 mg

   Tablets were prepared by tableting the above components with 150 mg of tablets according to the conventional method for producing tablets.

Formulation Example 2

* Preparation of capsule

   SJ-2030 50 mg

   Lactose 60 mg

   Dicalcium Phosphate 70 mg

   H, P, C 5 mg

   15 mg magnesium stearate

   The above ingredients were filled in 200 mg of light capsules according to the conventional method for preparing capsules to prepare capsules.

Formulation Example 3

* Preparation of Oral Solution

   SJ-2030 100 mg

   Caffeine anhydrous 30 mg

   Polysorbate 80 250 mg

   Di-sorbitol 1.5 g

   6 g of sugar

   1.5 g of methyl paraben

   Maleic acid 8.5 mg

   Alcohol 0.3L

   Appropriate amount of purified water

   The above components were filled and prepared with 30 ml of liquid according to a conventional method for preparing oral liquid.

Formulation Example 4

* Preparation of poultice (based on 10 g)

   SJ-2030 100 mg

   Menthol 80 mg

   Propylparaben 10 mg

   The above components were prepared as a poultice agent according to a conventional method for producing a poultice agent.

Formulation Example 5

 * Preparation of injections (based on 2 ml)

   SJ-2030 50 mg

   Sodium Chloride 18 mg

   Benzyl alcohol 45 mg

   Appropriate amount of distilled water for injection

   The above ingredients were filled into ampoules according to a conventional injection method to prepare injections.

       From the above results, the new lignan compound of the present invention, the compound of formula 1 (SJ-2030), showed an excellent hypoglycemic effect in the efficacy test, and even in the acute toxicity test, all other drugs (tolbuta) during oral and intraperitoneal administration. Much higher safety than amide). Therefore, SJ-2030 proved to be useful as a new oral hypoglycemic agent in terms of effectiveness and safety.

Claims (2)

(7R, 8R, 8'R, 9'R) -4, 7, 4 ', 9'-tetrahydroxy-3, 3'-dimethoxy- ^{1,3,5,1 ', 3', 5 '} -8 . 8 ', 9 . O. 9'-Lignan: (+)-Todolactol A (SJ-2030). Formula 1 (7R, 8R, 8'R, 9'R) -4, 7, 4 ', 9'-tetrahydroxy-3, 3'-dimethoxy- ^{1,3,5,1 ', 3', 5 '} -8 . 8 ', 9 . O. 9'-Lignan: Diabetes treatment containing (+)-todol lactola (SJ-2030) as an active ingredient. Formula 1