KR100615971B1 - Analogues of anticancer peptide synthesized and prepared from Gaegurin 5 - Google Patents

Abstract

The present invention relates to an anticancer peptide having high anticancer activity with a molecular size of less than half of that of openulin 5 through systematic peptide engineering of Gaegurin 5, an anticancer peptide isolated from domestic Rana rugosa , Specifically, the anticancer peptide derivatives synthesized and prepared from the shortest length of the open lipase 5 peptides isolated from the skin of domestic frogs have the shortest length among the known anticancer peptides, and have no cytotoxic and excellent anticancer activity. In addition, since hemolytic activity against human erythrocytes is low, it can be usefully used as an optimal anticancer agent safe for human body.

Mange frog, anticancer peptide, anticancer activity, anticancer agent

Description

Analogues of anticancer peptide synthesized and prepared from Gaegurin 5}             

Figure 1a is a side view of the structure of the peptide of the open mouth 5 (GGN5) in SDS micelles (cells),

Figure 1b is a view showing the top view of the N-terminal portion (G3 ~ V13) of the α-helix region of the opening 5 in the SDS micelle,

Figure 2 is a diagram showing a sequence list of the opening 5 and derivatives thereof,

3A and 3B show CD spectral analysis of A4W-GGN5N11 and V8W-GGN5N11, respectively.

4A is a diagram showing NMR analysis of A4W-GGN5N11 and V8W-GGN5N11,

4B is a diagram showing distance information of each remaining period of A4W-GGN5N11 and V8W-GGN5N11,

5a is a diagram showing the anticancer activity of paclitaxel and GGN5 against nine cancer cell lines,

Figure 5b is a diagram showing the anticancer activity of A4W-GGN5N11 and V8W-GGN5N11 against nine cancer cell lines.

The present invention relates to anticancer peptide derivatives synthesized and prepared from openulin 5.

Cancer is one of the incurable diseases that humanity must solve, and huge capital is invested in the development to cure it worldwide. In Korea, it is the number one disease death cause and more than 100,000 people per year It is diagnosed, and about 60,000 or more die. The growth rate of cancer patients is also reaching 10%. According to the National Cancer Center's statistics in 1999, there were 120,000 patients, with men having stomach cancer (24%), liver cancer (16%), lung cancer (16%), and colon cancer (10%), and women with stomach cancer (16 %), Cervical cancer (12%), breast cancer (15%) and colorectal cancer (10%). Carcinogens, which cause cancer, are smoking, ultraviolet rays, chemicals, foods, and other environmental factors. However, various causes are not only difficult to develop a therapeutic agent, but also effective effects vary depending on the site of occurrence. Currently, the substances used as therapeutic agents are extremely toxic and cannot selectively remove only cancer cells. Therefore, there is an urgent need for the development of low-toxic and effective anti-cancer agents to prevent cancer after treatment. .

Most anticancer agents are drugs that exhibit anticancer activity by interfering with various metabolic pathways of cancer cells and mainly inhibiting the synthesis of nucleic acids.

Research methods of anticancer agents for treating cancer have recently been investigated, such as searching for a direct cytotoxic substance to cancer cells, searching for a substance that modulates the immune ability of the living body, searching for a substance that inhibits the metastasis of cancer cells, and recently Various methods are in progress, such as searching for substances that inhibit angiogenesis. In addition, for the prevention of cancer, it is possible to use a substance that can effectively block various mechanisms of cancer development to prevent normal cells from transferring to cancer cells or to delay or restore the progression of carcinogenesis. Should be free of short or long term toxicity, be easy to take and inexpensive.

Currently used anticancer drugs can be broadly divided into biologics such as enzymes and vaccines, pure synthetic drugs, and natural products. Among them, anticancer drugs using genes, enzymes, vaccines, etc. are not in a practical stage and are used in chemotherapy. Many of the anticancer drugs developed by the drug have different pharmacological effects depending on the type of cancer (Gillman et al., The pharmacological Basis of therapeutics, Maxwell Macmillan. , 18 , p1202, 1986), It is pointed out as a city problem (Chung et al., J. Wonkwang Medical Sci. , 3 , pp 13-34, 1987). In addition, anti-cancer drugs effectively inhibit the growth of cancer cells, but sometimes become toxic to normal cells and become a major problem in cancer treatment, such as resistance to anti-cancer drugs as the cancer cells change during the process of growth, proliferation and metastasis. have. This is because most of the anticancer drugs have a molecular weight of 1,000 or less, and when the anticancer drugs are administered, they invade not only cancer tissues but also normal tissues and damage normal tissues, particularly tissue cells with active cell division, thus deteriorating bone marrow function and gastrointestinal tract. Various side effects such as disorders and alopecia will occur. In addition, because of its small molecular weight, it is easily excreted from the body by urine, which is caused by the large amount of drugs required for cancer treatment. Therefore, the toxicity to normal cells, excretion from the body, and the resistance of cancer cells with anticancer drugs are the biggest problems in the treatment of cancer, a problem that modern medicine must solve. Therefore, many scholars are conducting a number of studies to elucidate the mechanism of action of such toxicity (Yamazaki et al., Biosci, Biotech Biochem. , 56 (1) , p149, 1992; Tompson et al., Exp. Cell Res. , 41 , pp411-427, 1966; Ellem et al., Devel, Biol. , 118 , pp311-330, 1968), in recent years, as cell culture technology is rapidly developed, various cells are cultured and then various toxic substances are administered. As a result, studies are being actively conducted to investigate the mechanism of cytotoxicity at the cellular level, and development of anticancer drugs using herbal medicines and natural products has been continuously attempted to minimize side effects of anticancer drugs and to increase therapeutic effects.

In order to overcome the disadvantages of the conventional anticancer drugs, the present inventors found that the shortest length of the six antibiotic peptides (gaglin 1 to 6) was isolated from domestic frogs whose structures have already been identified by the present inventors. By confirming that the anticholinergic 5 with anticancer activity, based on this design and synthesis of a new anti-cancer peptide of a much smaller size, by identifying the excellent anticancer activity and hemolytic activity of the synthesized anticancer peptides, The invention has been completed.

An object of the present invention is to provide an anticancer peptide derivative having a novel structure designed and synthesized from an openrin 5 that has high anticancer activity and does not cause hemolysis when used in a human body, and an anticancer composition comprising the same.

In order to achieve the above object, the present invention provides an anticancer peptide derivative represented by F-L-G-W-L-F-K-V-A-S-K (A4W-GGN5N11) and F-L-G-A-L-F-K-W-A-S-K (V8W-GGN5N11) designed and synthesized from Openulin 5.

The anticancer peptide derivative is characterized in that it is synthesized by replacing a specific site of the peptide with tryptopan (Tryptopan).

The present invention also provides a pharmaceutical composition for the prevention and treatment of cancer, comprising the anticancer peptide derivative and the pharmaceutically acceptable carrier of the above designed and synthesized from an opening line 5, which has been found to have excellent anticancer activity.

Hereinafter, the present invention will be described in detail.

The anticancer peptide derivatives designed and synthesized from the openulin 5 of the present invention can be obtained by the following method.

The anticancer peptide derivatives of the present invention can be synthesized and prepared by engineering modification of the open chain 5 having the shortest length (24 residues) among the peptides isolated from the Korean frog, Om frog, (frog 1 to 6). Can be.

As described above, as a result of verifying the anticancer activity by removing the C-terminal residues of the openulin 5 one by one, the least residue having the anticancer activity was 13 residues. Since the usefulness of Trp in the anticancer peptide derivative development is expected from the derivative study of 4, a total of 11 Trp substituents of the openulin 5 may be synthesized by substituting the Trp residue for the 11 residue inactive segment instead of the 13 residue active segment of the openulin 5. Among the Trp substituents of the opening 5, the A4W-GGN5N11 in which Trp was introduced at position 4 and the V8W-GGN5N11 derivative in which Trp was introduced at position 8 were the parent molecules, and After confirming similar anticancer activity and hemolytic activity, other amino acids, preferably the hydrophobic residue Leu (leucine), hydrophilic to determine whether only Trp is specific at positions 4 and 8 In particular, a total of seven amino acid substituents of openulin 5 can be synthesized by substituting three amino acids of Phe (phenylalanine) having an aromatic ring such as Lys (lysine) and Trp having a positive ion.

In addition, the derivatives of the anticancer peptide of the present invention may be obtained by a chemical method using a peptide synthesizer, and may include a DNA or RNA sequence encoding an anticancer peptide, and may include a gene clone suitable for expressing the anticancer peptides. It may also be obtained by genetic recombination techniques that yield the above anticancer peptides expressed by genetic engineering and transplanted into suitable cells.

The present invention provides an anticancer peptide derivative obtained by the above method.

In addition, the present invention provides a pharmaceutical composition for preventing and treating cancer, comprising the anticancer peptide derivative and a pharmaceutically acceptable carrier thereof.

Among the anticancer peptide derivatives of the present invention obtained as described above, in order to find an optimal anticancer peptide having excellent molecular weight and stability, the anticancer activity and hemolytic activity of the anticancer peptide derivatives were examined. The same derivatives were found to exhibit anticancer and hemolytic activity similar to that of openulin 5.

GGN5N13: F-L-G-A-L-F-K-V-A-S-K-V-L

A4W-GGN5N11: F-L-G-W-L-F-K-V-A-S-K

V8W-GGN5N11: F-L-G-A-L-F-K-W-A-S-K

On the other hand, among the derivatives of openingulin 5, the following derivatives showed anticancer activity similar to that of openingulin 5, but it was confirmed that hemolytic activity was high.

AL4-GGN5N11: F-L-F-L-L-F-K-V-A-S-K

V8L-GGN5N11: F-L-G-A-L-F-K-L-A-S-K

W4,8-GGN5N11: F-L-G-W-L-F-K-W-A-S-K

In addition, other derivatives showed almost no anticancer activity against most cancer cell lines except for some cancer cell lines, and the anticancer peptide derivatives of the present invention had structural similarity to that of openulin 5. Derivatives having at least 11 of the 23 amino acid residues of openingulin 5 in order to have a structure, anticancer activity and hemolytic activity similar to that of openingulin 5 based on whether or not it has anticancer activity and hemolytic activity. Also, in order for the derivatives to have sufficient anticancer activity, it was confirmed that the derivatives had an affinity through Trp substitution at the position between the hydrophilic end and the hydrophobic start plane, and from these results, the optimal anticancer peptide for the development of anticancer drugs It was confirmed that the derivatives were A4W-GGN5N11 and V8W-GGN5N11. .

The pharmaceutical composition for preventing and treating cancer diseases of the present invention comprises 0.1 to 50% by weight of the compound based on the total weight of the composition.

Pharmaceutical compositions comprising the compounds of the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.

Pharmaceutical dosage forms of the compounds of the present invention may be used in the form of their pharmaceutically acceptable salts, and may be used alone or in combination with other pharmaceutically active compounds as well as in a suitable collection.

The pharmaceutical compositions comprising the compounds according to the present invention may be prepared in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories, and sterile injectable solutions, respectively, according to conventional methods. Can be formulated and used. Carriers, excipients and diluents that may be included in the composition comprising the compound include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate , Cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations may contain at least one excipient such as starch, calcium carbonate, sucrose, or the like. ) Or lactose, gelatin and the like are mixed. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

Preferred dosages of the compounds of the present invention depend on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, but may be appropriately selected by those skilled in the art. However, for the desired effect, the compound of the present invention is preferably administered at 0.0001 to 100 mg / kg, preferably at 0.001 to 10 mg / kg. Administration may be administered once a day or may be divided several times. The dosage does not limit the scope of the invention in any aspect.

The compounds of the present invention can be administered to mammals such as mice, mice, livestock, humans, and the like by various routes. All modes of administration can be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.

Hereinafter, the present invention will be described in detail by the following Examples and Experimental Examples.

However, the following Examples and Experimental Examples are only illustrative of the present invention, and the content of the present invention is not limited by the following Experimental Examples.

Example 1. Synthesis and Purification of Peptides

In order to synthesize an optimal anti-cancer peptide having an amphiphilic structure, an anti-cancer peptide, which is an anticancer peptide isolated from the epidermis of domestic scavenger, was used as a parent molecule and a new anticancer peptide having a much smaller size was standardized using standardized Fmoc-method. It was automatically synthesized using a solid phase method (Wellings, DA, and Atherton, E., Methods Enzymol., 289 , pp 44-67, 1997) using a peptide synthesizer (Model 90, Advanced Chemtech, Inc. United States of America). In other words, after adding 70 mM Resin (Resin Resin) to the reaction vessel (equivalent to 3 equivalents of amino acid for the resin, 2 equivalents of HOBT (1-Hydroxybenzotriazole) for the amino acid and 2 equivalents for the amino acid) DIC (1,3-Diisopropylcarbodiimide) was dissolved in an amino acid vessel (amino acid vessel) and dissolved in 10 ml of DMF (dimethylformamide), and then the resin of the reaction vessel was swelled using DMF. The Fmoc group of the resin was removed using 25% piperidine / DMF and then the resin was washed off with DMF, Dichloromethane (DCM) and Isopropanol (IPA). The amino acid dissolved in the amino acid container was transferred to the reaction container and reacted with the resin for 2 to 3 hours. In the case of attaching several amino acids, the above procedure was carried out in sequence to bind amino acids in sequence, and then the Fmoc group attached to the N-terminus of the last amino acid was removed using 25% piperidine / DMF. Remove the protecting group attached to the lysine or serine residue, add 20% of 10% Trifluoro acetic acid (DFA) / DCM to separate the synthesized peptide from the resin, and react for 4 hours. The reaction solution was distilled into a round flask. After all the solvents have evaporated, 10 ml of 20% acetonitrile (0.1% TFA) is added dropwise to dissolve again, and then filtered by centrifugation or a filter. The supernatant is lyophilized and reversed phase HPLC (WE J55B5) using a C-18 column is performed. , HITACHI, mobile phase: acetonitrile, fixed phase: C-18 resin, flow rate: 1 ml / min) was purified. For 45 minutes using acetonitrile / water mixed solvent containing 0.1% trifluor acetic acid at a flow rate of 1 ml / min, the fractions of which only the desired peptide was detected in the eluate were lyophilized, After obtaining the peptide, the molecular weight was measured using Mass Spectroscopy to obtain the results of A4W (MW: 1295.75) and V8W (MW: 1267.72). The sequence listing of the synthesized anti-cancer peptide derivatives of openulin 5 is shown in FIG. 2.

Experimental Example 1. Analysis of structure-function relationship of anticancer peptide

1-1. Circular Dichroism (CD)

The dried peptide powder obtained in Example 1 was precisely weighed and dissolved in various solvents at a concentration of 50 mM, and then the pH was adjusted to 4.0, and then used in a CD spectrum experiment. As solvent, 20 mM sodium acetate buffer (pH 4.0), 50% TFE / water, 5 mM DPC micelles and 10 mM SDS micelle solution were used. The CD signal was measured at 0.5 nm intervals from 250 nm to 190 nm at 20 mM using a JASCO J-720 spectrometer equipped with a temperature controller. At this time, the diameter of the measuring cell (container) was 0.2cm, the scan speed was set to 50nm / min, the bandwidth is 1nm, the response time (response time) was set to 4 seconds. Three separate measurements were taken for each experiment, and after subtracting the signal from the blank solvent, the mean value was converted to mean residue molar ellipticity by Equation 1 below.

(deg㎠·d·mol^-1)

(degcm2 · d · mol ⁻¹ )

(deg·㎠·dmol^-1)와

(mdeg) 는 각각 평균 잔기 몰 타원율과 특정파장 λ에서 측정된 CD 강도(3회 평균치)이고, ℓ, c, n 은 각각 측정용 셀(용기)의 지름(cm), 시료농도(mM), 시료 펩타이드의 잔기수이다.here

(deg · cm · dmol ⁻¹ ) and

(mdeg) is the CD residue (average of three times) measured at the average residue mole ellipticity and the specific wavelength λ, respectively, ℓ, c, n are the diameter (cm), the sample concentration (mM), The number of residues in the sample peptide.

As a result, both derivatives of A4W-GGN5N11 and V8W-GGN5N11 showed CD spectrum changes in the same pattern as the parent molecule, Openulin 5, under various solvent conditions (see FIGS. 3A and 3B).

1-2. Nuclear Magnetic Resonance (NMR) Analysis

Deuterium-labeled SDS was dissolved in water to make 300 mM SDS micelle solution, and peptide powder was dissolved therein to 3 mM. NMR measurement was performed at 40 ° C. with a Brooker DRX 500 spectrometer. The types of measurement spectra are the two-dimensional spectra of DQF-COSY, TOCSY (60 ms mixing time) and NOESY (200 ms mixing time). Solvent signal rejection is a selective pre-saturation method. NMR data were analyzed using the NMR Pipe / NMR Draw and NMR View program, and the DSS was used as a reference for the chemical shift of ¹ H. NMR analysis used a method standardized by Wuthrich (Wuthrich, K., NMR of Proteins and Nucleic Acids, John Wiley and Sons, New York, 1986). In other words, sequence-specific assignment was performed according to NOE connectivity on the NOESY spectrum after spin-system classification based on TOCSY and DQF-COSY.

As a result, more accurate structural information could be obtained from NMR analysis. As a result, both derivatives of A4W-GGN5N11 and V8W-GGN5N11 formed stable helix structure from Gly 3 or Leu 2 residues to Ser 10 or Lys 11 residues. It could be confirmed (see FIGS. 4A and 4B).

Experimental Example 2. Determination of anticancer activity of anticancer peptide

2-1. 50% inhibitory concentration (IC ₅₀ ) Measurement

In order to confirm the anticancer activity of the peptide derivative of the present invention obtained in Example 1, the microculture MTT method described in the literature (microculture MTT method, Angelina Quintero et al., J. Pharm Pharmaceut Sci., 2 (3)) , pp 108-112, 1999; Jin Yung Kim et al., Bioog.Med. Chem. Lett., 11 , pp2405-2408, 2001; Ashutosh K. Pathak et al., J. Am. Coll. Nutr., 21 ( 5) , pp416-421, 2002), and anti-cancer activity was verified by measuring IC ₅₀ of 9 cell lines.

The nine cell lines used in this experiment were: A498 (kidney), A549 (lung), HCT116 (colon), MCF-7 (breast), MKN45 (top), NCl-H630 (liver) , PC-3 (prostate), SK-MEL-2 (skin), and SK-OV-3 (ovarian cancer cell line). Cells were aliquoted into 96 well microtiter plates and incubated at 37 ° C. for 24 hours, and drug was added during exponential growth phase. Cells in the wells were seeded with and without dimethyl sulfoxide. After 3 days of incubation, 50 μl of 2 mg / ml MTT reagent was added to each well and incubated for 3 hours. The supernatant was removed, gently shaken to dissolve the remaining formazan crystals in the wells, and then 150ul of dimethyl sulfoxide was added. The wells without cells were blanked and the cells from the wells without drugs were used as a standard for cell viability, using a microplate reader at 570 nm at 570 nm. Tecan, Austria) absorbance was measured immediately. Inhibition rate on cell growth was calculated by the following equation (2).

% Inhibition on cell growth = {1- (uptake of treated cells / uptake of untreated cells)} X 100

The IC ₅₀ value of each cell was calculated as the amount of drug that caused a 50% reduction in uptake compared to untreated reference cells.

As a result, as shown in Table 1, FIGS. 5A and 5B, A4W-GGN5N11 and V8W-GGN5N11 both showed anticancer activity against various strains, and their anticancer activity also showed similar efficacy as that of the parent molecule, Openrin 5.

Cancer cell lines IC ₅₀ : mg / ml and (mM) Paclitaxel Native GGN5 A4W-GGN5 ^N11 V8W-GGN5 ^N11 A498 0.015 (0.017) 74.83 (54.1) 73.26 (56.5) 214.33 (169.1) A549 0.004 (0.0049) 78.96 (57.1) 106.27 (82.0) 418.15 (329.9) HCT116 0.002 (0.0024) 61.43 (44.4) 30.48 (23.5) 144.37 (113.9) MCF-7 0.002 (0.0019) 99.57 (72.0) 77.29 (59.6) 198.87 (156.9) MKN45 0.002 (0.0026) 18.98 (13.7) 82.70 (63.8) 142.72 (112.6) NCI-H630 0.021 (0.024) 22.67 (16.4) 75.92 (58.6) 129.54 (102.2) PC-3 0.015 (0.017) 23.74 (17.1) 123.81 (95.5) 174.15 (137.4) SK-MEL-2 0.006 (0.0072) 25.76 (18.6) 30.48 (23.5) na ^a SK-OV-3 0.004 (0.0050) 20.75 (15.0) 92.15 (71.1) 150.45 (118.7) ^a na: not available; 3 independent experiments do not yield reliable results

2-2. Hemolysis Measurement

3 ml of blood collected from healthy males were mixed with PBS (phosphate buffered saline (isotropic solution) 1: 1) (v / v), centrifuged to remove buffy coat and plasma, and again using physiological saline solution 3 Pure red blood cells were isolated by washing several times. It was suspended in 20 ml PBS and pre-incubated for 15 minutes in a water bath at 37 ℃. 10 ml of peptide solution per 190 ml of erythrocyte solution was mixed to prepare a mixed solution having a final peptide concentration of 100, 50, or 25 mg / ml, and then these were incubated for 30 minutes in a water bath at 37 ° C., followed by centrifugation and 100 ml of supernatant. Each solution was diluted in 1 ml of PBS, and the absorbance was measured at 550 nm (UV spectroscopy, Becknman, Germany). The% hemolysis was calculated from the ratio between the measured absorbance and the absorbance of the cell suspension treated with 0.2% Triton X-100.

As a result, as shown in Table 2, the hemolytic activity of A4W-GGN5N11 and V8W-GGN5N11 was also weak, similar to that of the parent molecule, Openin 5, and thus, it was confirmed that V8W-GGN5N11 was an A4W-GGN5N11. It showed less hemolytic activity.

In conclusion, it was confirmed that the V8W-GGN5N11 and A4W-GGN5N11 synthesized from the present invention 5 is useful as an excellent anticancer agent with reduced side effects such as hemolytic action.

Native GGN5 A4W-GGN5 ^N11 V8W-GGN5 ^N11 Peptide concentration Hemolysis% 100 µg / ml 6.78 11.9 6.76

Examples of the pharmaceutical composition of the present invention will be described, but the present invention is not intended to limit the present invention, but is only intended to be described in detail.

Formulation Example 1 Preparation of Powder

20 mg of peptide derivative of Example 1

Lactose 100 mg

Talc 10 mg

The above ingredients are mixed and filled in an airtight cloth to prepare a powder.

Formulation Example 2 Preparation of Tablet

 10 mg of peptide derivative of Example 1

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

After mixing the above components, tablets are prepared by tableting according to a conventional method for preparing tablets.

Formulation Example 3 Preparation of Capsule

10 mg of peptide derivative of Example 1

3 mg of crystalline cellulose

Lactose 14.8 mg

Magnesium Stearate 0.2 mg

According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare capsules.

Formulation Example 4 Preparation of Injection

10 mg of peptide derivative of Example 1

Mannitol 180 mg

Sterile distilled water for injection 2974 mg

_{Na 2 HPO 4 · 12H 2 O} 26 mg

According to the conventional method for preparing an injection, the amount of the above ingredient is prepared per ampoule (2 ml).

Formulation Example 5 Preparation of Liquid

20 mg of peptide derivative of Example 1

10 g of isomerized sugar

5 g of mannitol

Purified water

According to the conventional method of preparing a liquid solution, each component is added and dissolved in purified water, lemon flavor is added to the mixture, and then the above ingredients are mixed, purified water is added to adjust the total amount to 100 ml, and then filled in a brown bottle. The solution is prepared by sterilization.

As described above, the anticancer peptide derivatives designed and synthesized from the openulin 5 of the present invention not only show excellent anticancer activity in 9 cancer cell lines, but also have very low hemolytic activity against human red blood cells. All side effects due to anticancer drugs can be significantly reduced, and since it has the shortest length among anticancer peptides known so far, it can be usefully used as an optimal anticancer agent since it is very advantageous in absorption and drug transport.

Claims (6)

A pharmaceutical composition for preventing and treating cancer, comprising an anticancer peptide derivative represented by F-L-G-W-L-F-K-V-A-S-K (A4W-GGN5N11) and a pharmaceutically acceptable carrier.        A pharmaceutical composition for preventing and treating cancer, comprising an anticancer peptide derivative represented by F-L-G-A-L-F-K-W-A-S-K (V8W-GGN5N 11) and a pharmaceutically acceptable carrier. delete An antimicrobial pharmaceutical composition comprising the antibiotic peptide derivative of claim 1 and a pharmaceutically acceptable carrier. delete A pharmaceutical composition for antimicrobial comprising the antibiotic peptide derivative of claim 2 and a pharmaceutically acceptable carrier.

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