KR100295269B1 - New Ichinomicin derivatives for cancer treatment and preparation method thereof - Google Patents

Abstract

The present invention relates to the synthesis of novel Ichinomycin derivatives using Ichinomycin known as anticancer agents. Ichinomycin is a substance in Phase II of clinical trials and has many problems to be used as an anticancer agent because of its high cytotoxicity to normal cells and low solubility in polar solvents.

Sulfated derivatives and sulfonium derivatives of Ichinomycin synthesized according to the present invention solve the problem by improving the solubility in polar solvents and reducing the cytotoxicity to normal cells since the introduction of polar groups and derivatization in the form of chlorides. It was. After synthesizing the compounds of the present invention, pure anti-separation and purification were performed to determine anticancer activity. The sulfoxide, disulfoxide, and sulfone synthesized through the oxidation of sulfur element did not significantly improve the anticancer activity, but only slightly from sulfoxide. A reduction in toxicity was obtained. However, the compound obtained through the sulfonium salt synthesis reaction of sulfur element was able to obtain a result of more than 100-fold reduction in cytotoxic ability against normal cells while maintaining anticancer activity.

Description

Novel Ichinomycin Derivatives Treating Cancer and Methods for Preparing the Same

The present invention relates to the synthesis of Ichinomycin derivatives using Echinomycin known as an anticancer agent.

Since Ichinomycin is known as a new substance in 1974, it is being studied for use as an anticancer activity, structural analysis, DNA binding site and anticancer agent. However, due to its low solubility in polar solvents, it has many problems to be used as an anticancer agent. There are about 10 derivatives of Ichinomycin and Triostin which are separated from nature, but synthetic derivative is the only derivative of Triostin which is a natural product rather than synthetic derivative. There is a lot of research on the synthesis method. However, the tandem derived from triostin is a compound having a disulfide as a basic structure and does not have a hydrophilic group, so it is difficult to fundamentally solve the problems of the existing ichiomycin. The general structural formula of Ichinomicin is shown below.

The present invention focuses on the fact that a sulfur compound is contained in the structure of ichiomycin, thereby improving polarity through oxidation of sulfur and synthesizing sulfonium salts on sulfur elements to complete the present invention. An object of the present invention is to improve the solubility in polar solvents and to reduce the cytotoxic ability to normal cells by the synthesis of the sulfated derivatives and sulfonium derivatives in the form of a polar group or a derivative of chloride.

The present invention is synthesized in the chloride form to maximize the polarity of the Ichinomycin chloride represented by the following formula (1), trialkyloxonium tetrafluoroborate or Meerwein salt (Meerwein salt) Reagents can be used to synthesize chlorides of Ichinomycin, or chlorides of Ichinomycin can be easily synthesized using a combination of silver perchlorate (AgClO ₄ ) and methyliodine (MeI).

(One)

In addition, the sulfur oxides of ichiomycin represented by the following structural formula (2) are chlorinated by using reagents such as sodium iodide oxide (NaIO ₄ ) _, metachloroperbenzoic acid (m-CPBA) and benzene iodide as sulfur compounds. Sulfur oxides can be synthesized. Since the sulfation reaction cannot selectively form only sulfoxide or sulfone, each component such as sulfoxide, disulfoxide and sulfone can be separated and purified in pure form using column chromatography or preparative liquid chromatography to carry out anticancer activity. Can be measured.

(2)

Hereinafter, the content of the present invention will be described in detail in Examples and Test Examples. However, these are provided only to facilitate the description of the present invention, and the technical scope of the present invention is not limited thereto.

Example 1 Synthesis of Ichinomycin Sulfur Oxide

Dissolve 1.1 g of purely purified Ichinomycin in 70 ml of purified methylene chloride solvent and slowly add dropwise 0.23 g of metachlorobenzoic acid at 0 ° C., react for 24 hours at room temperature, and then add 100 ml of water to the reaction. . The synthesized sulfur oxides were extracted three times with 100 ml of methylene chloride solvent and the methylene chloride layer was washed with saturated brine and then water was removed using magnesium sulfate. When the methylene chloride is concentrated, 0.7 g of Ichinomycin sulfur oxide is obtained as a mixture, and fractionated using preparative thin layer chromatography (TLC), followed by pure separation using preparative liquid chromatography. Preparative conditions are purely separated by reverse phase column (Ooctadecylsilane, ODS) using a solvent mixed with acetonitrile: water = 70: 30, the pure form of Ichinomycin sulfur oxide is 80mg sulfoxide, disulfoxide 80 mg and sulfone obtained 210 mg. Each purely isolated compound was analyzed by mass, FT-IR and FT-NMR. Ichinomycin sulfoxide was found to have a molecular weight of 1117 (M + 1) in electro spray ionization (ESI) -Mass. , FT-IR confirmed the characteristic band of sulfoxide, 1036cm ^-1 , and the FT-NMR also showed that the methyl group changed from 2.0ppm to 2.43ppm compared with the conventional ichiomycin. Ichinomycin disulfoxide was confirmed to have a molecular weight of 1133 (M + 1) in ESI-Mass, and the FT-NMR also showed that the methyl group was changed from 2.0 ppm to 2.58 ppm in comparison with conventional ichiomycin. Ichinomycin sulfone was found to have a molecular weight of 1133 (M + 1) in ESI-Mass and 1261 cm ^-1 , a characteristic band of sulfone in FT-IR. In FT-NMR, the methyl group was changed from 2.0 ppm to 2.90 ppm compared with the conventional ichiomycin.

Example 2 Synthesis of Ichinomycin Sapluoroborane Chloride

1.1 g of purely purified Ichinomycin was dissolved in 60 ml of purified dichloroethane solvent, and 0.82 ml of triethyloxonium safuluroborane was slowly added dropwise at room temperature and reacted at room temperature for 4 hours to form Ichinomycin chloride. It can be confirmed that the deposit is formed. The reaction mixture was obtained by depositing the precipitate using a centrifuge, and then washed with unreacted ichiomycin by using chloroform solvent. The compound was separated purely using a reverse phase column and preparative liquid chromatography with a solvent mixed with preparative TLC and acetonitrile: water = 70: 30. Ethyl safuluoroborane chloride of Ichinomycin was found to have a molecular weight of 1129 (M + 1) in ESI-Mass and methyl safuloboroborane chloride was found to have a molecular weight of 1115 (M + 1) in ESI-Mass.

Example 3 Synthesis of Ichinomycin Methyl Perchlorate Chloride

Dissolve 200 mg of purely purified Ichinomycin in 10 ml of purified dichloroethane, and then slowly add dropwise 45 mg of perchloric acid with 400 mg of methyl iodine at room temperature and react for 24 hours at room temperature to form Ichinomycin chloride and silver iodide. It can be seen that (AgCl) deposits are produced. After the reaction is completed, remove the silver iodide deposit using a centrifuge, wipe off the unreacted Ichinomycin using chloroform solvent, dissolve it in acetonitrile, extract and remove water using magnesium sulfate. Concentration of the acetonitrile (CH ₃ CN) solvent gives a mixture of about 200 mg, which can be separated and purified in pure form using an ODS column and preparative liquid chromatography with a solvent mixed with acetonitrile: water = 70: 30. Can be. As a result of the separation and purification, 140 mg of Ichinomycin methyl perchlorate chloride in pure form was obtained. The structure was analyzed by mass, FT-IR and FT-NMR. Ichinomycin methyl perchlorate chloride had a molecular weight of 1115 (positive) in ESI-Mass. ), Identified as 102 (negative) and is in good agreement with the precalculated molecular weight. In FT-NMR, the methyl group changed from 2.0ppm to 2.17ppm compared with the conventional Ichinomycin. In particular, methyl group alcohol was produced by NOE (Nnuclear overhouse effect) effect confirmed by 2D nuclear magnetic resonance spectroscopy (2D NMR NOESY). It was confirmed that it was adjacent to the poinium salt.

Test Example 1 In Vitro Anticancer Activity of Ichinomycin Derivatives

As a result of measuring the anticancer activity of the in vitro toxin by purely separating and purifying the compound synthesized in the above example, sulfoxide, disulfoxide, and sulfone synthesized through the oxidation reaction of sulfur element did not significantly improve the anticancer ability. Although the toxicity was slightly reduced only in the sulfoxide, the synthesis compound of the sulfur element synthesized sulfonium salts resulted in more than 100-fold reduction in the cytotoxic activity against normal cells while maintaining anticancer capacity. there was. The anticancer mechanism and anticancer activity of the novel sulfonium salt ichiomycin derivatives are different from those of ichiomycin. It showed similar anticancer activity compared to nomycin. Details are shown in Table 1 below.

Table 1.In Vitro Anticancer Activity of Ichinomycin Derivatives

Test Example 2 In vivo Anticancer Activity of Ichinomycin Derivatives

In vivo anticancer activity of the mice was measured using the ichiomycin derivative synthesized in the example, and is shown in detail in Table 2 below. As shown in Table 2 below, the chloride of Ichinomycin (particularly methyl sulfonium chloride) has a higher survival rate than conventional Ichinomycin. It seems possible.

Table 2. In vivo Anticancer Activity of Ichinomycin Derivatives

compound N ^a P388 (ip-ip) ^b B16 (ip-ip) ^b mg / kg MST (%) ^c ± SD (Day) mg / kg MST (%) ^c ± SD (Day) Control group ^e 10 26 ± 2 18 ± 3 Methylsulfonium 88666 10.250.1250.06250.03125 35 ^d ± 1234 ^d ± 1233 ^d ± 732 ± 331 ± 4 10.250.1250.06250.03125 28 ^d ± 1326 ^d ± 225 ^d ± 324 ^d ± 523 ± 2 Sulfoxide 8 One 27 ± 4 One 20 ± 4 Disulfoxide 8 One 31 ± 1 One 17 Sulfone 8 One 25 ± 5 One 17 Methyl Safulobororon 6 One death 0.25 20 Ethyl safuluoroboron 8 One death 0.25 20 Ichinomicin 68886 0.10.050.0250.01250.00625 27 ± 426 ± 533 ± 332 ± 231 ± 2 0.10.050.0250.01250.00625 21 ± 223 ^d ± 625 ^d ± 524 ^d ± 223 ± 2

^a N: number of mice

^b After cancer cells were transferred intravenously to Mice (BDF1), anticancer activity was measured by intravenous injection of selected compounds for 1 to 9 days.

^c MST: survival days of treated mice

^d P <0.05 Determination of Anticancer Capacity by Kaplan-Meier Method

^e Control group: Mice that only metastasized cancer but did not take any action

Chloride of Ichinomycin prepared by the present invention (particularly methyl sulfonium chloride) in vivo experiments using mice significantly increased the survival rate compared to conventional Ichinomycin, especially the dose to increase the toxicity to normal cells It is possible to control the dosage and to improve the polarity by oxidative reaction with sulfur compound contained in Ichinomycin, and to obtain sulfonium salt by combining sulfur element with pure element. The cytotoxic ability against the 100 times reduced to solve the conventional problem.

Claims (3)

Ichinomycin derivative which is a cancer treatment agent represented by following General formula (1) as an ichiomycin chloride derivative. (One) However, in the formula (1), X represents halogen, sulfate, and perchlorate capable of synthesizing a sulfonium salt. A method for producing an inomycin derivative, which is a cancer therapeutic agent, using a compound capable of producing tetraalkyl oxonium safuluroborane, silver perchlorate and chloride in synthesizing the alkyl sulfonium salt of formula (1). Cancer therapeutic agent, characterized in that the derivative of Ichinomycin represented by the formula (1) or (2) as an active ingredient.