KR20070047576A - Use of the gonadotropin-releasing hormone ii antagonists and its analogues - Google Patents

Abstract

The present invention is directed to tryptorelix-1 and its analogs, which are inhibitors of gonadotropin-releasing hormone-2 (GnRH-2) action. According to the present invention, tryptorelix-1 and its analogs specifically act on prostate cancer cells to induce apoptosis, and thus can be used for the treatment and prevention of prostate cancer.

Gonadotropin releasing hormone-2, tryptorelix-1, inhibitors, analogs, prostate cancer, cell death, prostate cancer treatment

Description

Use of gonadotropin-releasing hormone II agonists and analogues thereof {USE OF THE GONADOTROPIN-RELEASING HORMONE II ANTAGONISTS AND ITS ANALOGUES}

1 is a diagram showing the death of prostate cancer cells DU-145 cells according to the treatment time of Tryptorelix-1.

Figure 2 is a diagram showing the apoptosis effect of prostate cancer cells, breast cancer cells, uterine cancer cells according to tryptorelix-1 treatment.

Figure 3a is a diagram showing the effect of Tryptorelix-1 and its analogs on prostate cancer killing.

FIG. 3B shows the amino acid sequence of Tryptorelix-1 and its analogs. FIG.

Figure 4a is apoptosis effect according to the serum concentration of prostate cancer cell culture.

Figure 4b is the effect of apoptosis according to the concentration of tryptorelix-1 when cultured in 2% serum prostate cancer cells.

FIG. 5A is a diagram in which prostate cancer cells are treated with Setrorelix and Tryptorelix-1 and stained with a DAPI fluorescence staining method.

Figure 5b is a diagram showing the treatment of Setrorelix and Tryptorelix-1 to prostate cancer cells and DNA fractions observed on electrophoresis.

5c is a diagram in which prostate cancer cells are treated with Setrorelix and Tryptorelix-1 and DNA fractions are stained by TUNEL staining in cells.

Field of technology

The present inventors have developed and applied US patents for agents and inhibitors that selectively act on gonadotropin-releasing hormone-2 (GnRH-2) (Seong JY, Kwon HB (10/28/2004) US patent application: Agonists and antagonists of gonadotropin-releasing hormone-2, and use according.10 / 513,083). Among the agents and inhibitors of the present invention, tryptorelix-1 (Trptorelix-1) has a direct and specific effect on prostate cancer cell death, so that tryptorelix-1 and its analogs can be used to treat prostate cancer. To present.

Prior art

The prostate gland is a male reproductive organ that produces semen, which is essential for the movement of sperm and is an organ that is constantly growing by male steroid hormones. In men, prostate disease develops after age 40, and prostate-related diseases are prostatitis, benign prostatic hyperplasia (BPH), and prostate cancer. Prostate cancer can be divided into early hormone dependent (reactive) prostate cancer and late hormone refractory prostate cancer. Hormone-dependent prostate cancer can be delayed through surgery or through powerful inhibitors of the male hormone synthesis inhibitor finasteride (an inhibitor of 5α-reductase), or gonadotropin-releasing hormone-1 (GnRH) (Eisenberger MA et al., J Clin Oncol 4: 414-424, 1986). However, after a period of time, these hormone-dependent prostate cancers develop into late-hormone refractory prostate cancer and no longer act as hormone therapy. Hormonal refractory prostate cancer has a high mortality rate due to active cancer metastasis to lymph glands and bones, but no suitable treatment is currently developed (Navarro et al., J Steroid Biochem Mol Biol 81: 191-201, 2002). Therefore, if there is a chemotherapeutic agent for late prostate cancer, it is expected to be very useful.

GnRH-1 synthesized in the hypothalamus plays an important role in regulating steroid concentration in vivo. Continuous treatment of GnRH-1 at high concentrations or treatment with GnRH-1 antagonists lowers the function of the GnRH-1 receptor present in the pituitary gland, ultimately reducing the function of the gonads and steroid concentrations in vivo. Is lowered. The gonad-inhibiting action of GnRH-1 is called " medical castration " and this method is very useful for the treatment of steroid hormone-dependent prostate cancer (Comaru-Schally et al., J Clin Endocrinol Metab 83: 3826-3831, 1998 Cook T and Sheridan WP, Oncologist 5: 162-810, 2000). In addition to these indirect effects, direct inhibition of GnRH-1 in prostate cancer cells was observed (Moretti RM et al., J Clin Endocrinol Metabol 81: 3930-3937, 1996; Limonta P Endocrinology 140: 5250-5256, 1999). This suggests the possibility that GnRH can act directly on cancer cells. However, the effect of GnRH-1 is a growth inhibitory effect of prostate cancer cells and does not induce cell death.

We have developed agonists and inhibitors of GnRH-2 and showed that they act more effectively than GnRH-2 on GnRH-2 receptors found in non-mammalian and monkeys (Maiti K et al., Mol Cells 16: 173-). 179, 2003; Wang AF et al., Mol Cell Endocrinol 209: 33-42, 2003). In the human genome, the gene for the GnRH-2 receptor found in monkeys has been identified as a pseudogene, suggesting that there is no functional GnRH-2 receptor in humans (Morgan K et al., Endocrinology 144: 423-). 436, 2003) and the results of other researchers, including the present inventors, have suggested that there are other forms of GnRH-2 receptors separate from these genes (Kang SK et al., Endocirnology, 142: 182-192, 2001; Chen). A et al., Nat Med 8: 1421-1426; Emons G et al., Endocr Relat Cancer 10: 291-299; Maiti K et al., Supra, 2005).

The inventors have identified a protein that binds GnRH-2 to hormone-refractory prostate cancer cells DU-145, ALVA-41, PPC-1 and TSU-Pr1 cells, which have a molecular weight of about 80 kDa. These cells increased the intracellular calcium concentration in response to GnRH-2, and when treated with GnRH-2 agonist tryptorelix-1, it was confirmed that this reactivity disappeared, GnRH-2 to these cells It is shown that there is a receptor. In addition, the present inventors have completed the present invention by clarifying that these cancer cells are killed by tryptorelix-1 but not by cetrorelix, a GnRH-1 agonist.

As such, the main object of the present invention is to provide a peptide that induces prostate cancer cell apoptosis as a gonadotropin releasing hormone II agonist.

Another object of the present invention to provide a prostate cancer therapeutic agent comprising the peptide.

The Tryptorelix-1 peptide, a GnRH-2 agonist of the present invention, is "Ac-D2Nal- (4Cl) D-Phe-D-3Pal-Ser-Tyr-D-Cit-Trp-Tyr-Pro-DAlaNH ₂ " It has the same chemical structure as (SEQ ID NO: 1). Herein, the modified amino acid is a compound name widely used in the art, and specifically Ac-D-Nal is an acetyl group substituted with D-alanine having a naphthyl group substituted at a beta-carbon position (β- ( 2-naphthyl-D-Ala)), (4Cl) -D-Phe is D-alanine substituted with a pyridyl group at the beta-carbon position linked to position 3 of the pyridine ring, and D-Cit is the D of citrulline. It is an isomer. Tryptorelix was prepared by modifying Setrorelix (Cetrorelix, FIG. 3b), which is well known as an inhibitor of GnRH-1. Setrolelix is an inhibitor of action by replacing amino acids 1, 2, 3, and 6 of GnRH-1 with amino acid derivatives, and shows high binding capacity with GnRH-1 receptor but does not activate the receptor. Tryptorelix-1 was synthesized by substituting the seventh and eighth amino acids of Setrorelix. We have higher selectivity for GnRH-2 receptors found in monkeys and non-mammalians than trylorellix-1 and more effectively inhibit the activity of GnRH-2 receptors (Mait K et al., Supra, 2003; Wang AF et al., Supra, 2003). The prostate cancer cell apoptosis inducing peptide of the present invention may exhibit apoptosis activity if it has at least 80% homology with the amino acids constituting the tryptorelix of SEQ ID NO: 1.

GnRH-1 action and inhibitors have been used to treat prostate, breast and ovarian cancer cells caused by steroids (Schally AV, Peptides 20: 1247-1262, 1999; Grundker C, et al., Eur J Endocrinol 146: 1-14, 2002). Both agonists and inhibitors of GnRH-1 reduce or inhibit the number of pituitary GnRH-1 receptors, thereby inhibiting steroid secretion in the testes or ovaries. Lowering steroid levels in the body ultimately inhibits the growth of tumors or cancer cells formed in the prostate, breast, or ovaries. In recent years, the expression of GnRH-1 receptors in these cancer cells has been shown to study the direct effects of GnRH (Moretti RM et al., J Clin Endocrinol Metabol 81: 3930-3937, 1996; Limonta P Endocrinology 140: 5250 -5256, 1999).

However, no specific study on the treatment of endocrine-related cancer cells by GnRH-2 has been reported. The inventors of the present invention describe five prostate / urinary cancer cells derived from humans, DU-145 (Moretti RM et al., Supra, 1996), ALVA-41 (Nakhla AM and Rosner W, Steroids 59: 586-589), PPC-1 (Brothman AR et al., Cancer Genet Cytogenet 37: 241-248, 1989), TSU-Pr1 (Iizumi T et al., J Urol 137: 1304- 1306), LNCaP (More RM et al., Supra Thesis, 1996) investigated the reactivity to GnRH-2 in cells. The DU-145, ALVA-41, PPC-1, and TSU-Pr1 used in the experiment were hormone refractory prostate cancer cells, while LNCaP cells were hormone reactive prostate cancer cells. All hormone refractory prostate cancer cells except LNCaP cells were observed to increase the concentration of intracellular Ca ²⁺ by GnRH-2 treatment. Responses such as Ca ²⁺ increase were not observed in LNCaP cells and breast cancer cells MCF-7 cells and kidney cells HEK293 cells. These results suggest that prostate cancer cells have proteins or receptors that respond to GnRH-2. We have identified GnRH-2 binding protein in hormone-resistant prostate cancer cells using GnRH-2 radiolabeled with Iodine ¹²⁵ . In addition, tryptopelix-1, a GnRH-2 inhibitor, has been shown to inhibit cell division in prostate cancer cells and ultimately induce cell death. Although LNCaP cells, which are hormone-reactive prostate cancer cells, did not respond to GnRH-2 as increased Ca ²⁺ , there was a possibility of GnRH-2 binding protein when examined with GnRH-2 radiolabeled with Iodine ¹²⁵ . Inhibition of cell division was observed by treatment with Tryptorelix-1. These results thus suggest the possibility that tryptorelix-1 can be used for the treatment of hormone dependent (reactive) and hormone refractory prostate cancer cells.

Tryptorellix-1 and its analogs of the present invention are useful as prostate cancer therapeutic agents because they act directly and specifically on the killing of prostate cancer cells. Accordingly, the present invention provides a pharmaceutical composition for treating prostate cancer containing tryptorelix-1 and its analogs as an active ingredient.

In order to achieve the other object of the present invention, the present invention provides a DNA or RNA oligonucleotide comprising a nucleotide sequence encoding the amino acid sequence of the tryptorelix-1 peptide that induces prostate cancer cell apoptosis of the present invention .

The pharmaceutical compositions of the invention can be formulated in a variety of oral or parenteral dosage forms. Formulations for oral administration include, for example, tablets, pills, light and soft capsules, solutions, suspensions, emulsifiers, syrups, granules, elixirs, and the like, in addition to the active ingredients, these formulations may contain diluents (e.g., lactose). , Dextrose, sucrose, mannitol, sorbitol, cellulose and / or glycine), glidants such as silica, talc, stearic acid and its magnesium or calcium salts and / or polyethylene glycols. Tablets may also contain binders such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and / or polyvinylpyrrolidine, optionally starch, agar, alginic acid or its Disintegrants or boiling mixtures such as sodium salts and / or absorbents, colorants, flavors, and sweeteners.

 In addition, pharmaceutical compositions containing tryptorelix-1 and its analogs as active ingredients may be administered parenterally, and parenteral administration may be by subcutaneous injection, intravenous injection, intramuscular injection or intrathoracic injection. All. At this time, the tryptorelix-1 and its analogs may be mixed in water with a stabilizer or buffer to formulate into a parenteral formulation, and then prepared as a solution or suspension, which may be prepared in a unit dosage form of ampoules or vials. have.

The pharmaceutical composition may contain sterile and / or preservatives, stabilizers, emulsifiers or emulsifiers, auxiliaries such as salts and / or buffers for the control of osmotic pressure, and other therapeutically useful substances, and conventional methods of mixing, It may be formulated according to the granulation or coating method.

In order to introduce a peptide of the present invention into a cell, a protein transduction domain (PTD) may be fused with the peptide to generate a fusion protein. The protein transport domain (PTD) can use any PTD known in the art.

Tryptorellix-1 and its analogs as active ingredients are in an amount of 0.01 to 500 mg / kg body weight per day, preferably 0.1 to 50 mg / kg body weight per day for mammals, including humans. It may be administered once or in divided oral or parenteral routes. The dosage may be appropriately selected depending on the age, sex and severity of the patient.

Hereinafter, the present invention will be described in more detail with reference to Examples. However, the following examples are only for illustrating the present invention, and the scope of the present invention is not limited thereto.

Example

Example 1 Apoptosis Effect by Tryptorelix-1 Treatment

Prostate cancer cells DU-145 and LNCaP cells were distributed from Seoul National University Cell Line Bank (Seoul Korea). Another prostate cancer cell, ALVA-41, was cultured with the permission of Dr. Rosner W, the original producer (Nakhla AM & Rosner W, Steroids 59: 586-589, 1994). DU-145 cells were cultured in DMEM broth (Invitrogen 11960-044) containing 10% heat-inactivated fetal bovine serum. One day before the experiment, 1 × 10 ⁴ cells were transferred to each well of a 24-well plate and incubated in DMEM containing 0.1% fetal bovine serum. One day after the cells were transferred, tryptorellix-1 was treated at 10 μM concentration and cells were counted after 6, 12, 24, 48, 72 and 96 hours, respectively (FIG. 1). The number of cells increased with time in the cell group not treated with Tryptorelix-1, while the number of cells significantly decreased after 24 hours in the group treated with Tryptorelix-1, and rarely observed after 96 hours. (FIG. 1).

Example 2: Killing Effect of Other Cells by Tryptorelix-1 Treatment

To compare the effects of tryptorelix-1 on other cells, breast cancer cells (MCF-7) (ATCC HTB-22), uterine cancer cells (HeLa) (ATCC CCL-2) and another prostate cancer cell, ALVA-41 ( Obtained from Dr. Rosner W.) and LNCaP cells were treated with tryptorelix-1. DU-145 and ALVA-41 cells are late prostate cancer cells and are male hormone refractory prostate cancer cells, and LNCaP cells are male hormone reactive prostate cancer cells. All cells were planted by 1 × 10 ⁴ numbers in 24-well plates and incubated in DMEM containing 0.1% fetal bovine serum. Tryptorelix-1 was treated for 24 hours at 10 μM concentration and the cells were counted. Prostate cancer cells DU-145, ALVA-41, LNCaP cells were killed more than 90% by 24 hours treatment, whereas apoptosis was not observed in MCF-7 and HeLa cells. These results indicate that tryptoelix-1 acts specifically on prostate cancer cells (FIG. 2). Tryptorelix-1 also acted on LNCaP cells, suggesting that tryptorelix-1 can be used to treat hormone-reactive prostate cancer cells as well as hormone-resistant prostate cancer cells.

Example 3: Apoptosis Effect by Treatment of Tryptorelix-1 and Its Analogues

1 × 10 ⁴ DU-145 cells were transferred to each well of a 24-well plate and incubated for one day in DMEM containing 0.1% fetal bovine serum. Cells were treated with Tryptorelix-1 and analogs thereof at 10 μM concentration for 24 hours and the cells were counted (FIG. 3A). The amino acid sequence of Tryptorelix-1 and its analogs is shown in Figure 3b. Among Tryptoelix-1 analogs, N3 showed similar effects to Tryptorelix-1, and N4 and N5 were observed to induce cell death at 50% level. Other analogues did not show the same effect as tryptorelix-1, and serorelix, a GnRH-1 agonist, was found to have minimal cell death effects.

Example 4 Influence of Serum Concentration on the Cell Killing Effect by Tryptorelix-1

1 × 10 ⁴ DU-145 cells were transferred to each well of a 24-well plate and cultured in DMEM containing fetal bovine serum, 0.1, 0.5, 1.0, 2.0, 5.0 and 10.0 wt%, respectively. Cells were treated with Tryptorelix-1 at 10 μM concentration daily for 5 days and the cells were counted (FIG. 4A). In the experimental group not treated with tryptorelix-1, the cell number increased with increasing serum concentration. In the experimental group treated with Tryptorelix-1, the number of cells was significantly reduced. In particular, in the experimental group cultured at 0.1, 0.5, and 1.0% of serum concentrations, about 95% of the cells were killed by Tryptoelix-1 treatment at about 2 days, and no viable cells were observed at 5 days. In addition, in the experimental group cultured in 2.0 and 5.0% serum, about 95% of the cells were killed by tryptorelix-1 treatment in about 3 days. In the experimental group incubated with 10% serum, an effect of about 82% cell number reduction was observed on the 5th day by trytrellix-1 treatment. These results indicate that the effect of tryptorelix-1 is related to serum concentration and that the higher the serum concentration, the less the effect of tryptorelix-1. Cultures were cultured in 2% serum to investigate the cell division inhibitory effect of tryptorelix-1, treated daily at concentrations of 0.001, 0.01, 0.1, 1 and 10 μM for 4 days and observed cell number changes. The cell number reduction effect was not observed when the concentration was 0.001 and 0.01 μM, but the concentration was decreased depending on the concentration of 0.1, 1 and 10 μM. These results show that tryptorelix-1 has a cell division inhibitory effect at concentrations of 0.1 and 1.0 μM.

Example 5: Apoptosis of Tryptorelix-1 through Apoptosis Induction

To determine the pathogenesis of the cell death effect by tryptorelix-1, the cells were treated with serorelix, or tryptorelix-1, and the nucleus was 4 ', 6-diamidino-2. Staining with -phenylindole (4 ', 6-Diamidino-2-phenylindole) (DAPI) (Sigma D8417) (FIG. 5A), obtaining DNA from cells and observing DNA fractionation (FIG. 5B), or in the cell nucleus DNA fractions were observed by terminal deoxynucleotidyl transferase-mediated deoxyuridine 5-triphosphate (dUTP) nick-end labeling (TUNEL) staining (FIG. 5c). 8 x 10 ⁴ cells were incubated under 2% serum in 6-well plates with poly-L-lysine-coated glass cover slip for DAPI staining. Tryptorelix-1 or Setrorelix was treated at 10 μM concentration for 3 days and cells were washed. Cells were fixed for 15 minutes in phosphate buffered saline (PBS) with 2% paraformaldehyde (Invitrogen 21600-069) and treated with 0.2% Triton X-100 (tradename). It was then immersed in DAPI solution (0.1% Triton X-100, 2 mM MgCl ₂ , 0.1 M NaCl, 100 mM PIPES, 1 μg / ml DAPI, pH 6.8) for 5 minutes, washed again, and observed by fluorescence microscope. In the cells treated with Tryptorelix-1, the nuclei were fractionated, and in some cells, nuclei were completely extinguished (FIG. 5A). To observe DNA fragmentation, 1 × 10 ⁶ cells were incubated with 2% serum culture in 100-mm dishes and treated with Tryptorelix-1 or Setrorelix once for 3 days. After the cells were collected, lysis buffer (20 mM Tris-HCl, pH 8.0, 10 mM EDTA, 0.5% Triton X-100) was added and stored on ice for 30 minutes. The cells were then homogenized and centrifuged at 15,000 rpm for 15 minutes. The supernatant was collected and mixed with phenol-chloroform-isoamylalcohol in a 1: 1 volume ratio and centrifuged again to collect the supernatant. The supernatant was then mixed with 70% ethanol and stored for one day in a -70 ° C freezer. DNA pellets were obtained by centrifugation, and RNase was treated for 15 minutes at 37 ° C, followed by electrophoresis. Electrophoresis showed that DNA ladders were cut off uniformly in the Tryptorelix-1 treatment group, but DNA ladders were not observed in the Setrorelix treatment group and the control group (FIG. 5B). In order to observe the DNA fraction on the cells, cells were cultured under the same conditions as when DAPI staining was performed, and then treated with Tryptellix-1 or Setrorelix for 3 days. Cells were fixed in PBS with 4% paraformaldehyde and treated for 15 minutes in methanol with 3% H ₂ O ₂ . Next, 0.1% sodium citrate solution containing 0.1% Triton X-100 was treated for 2 minutes, and TUNEL reaction solution (Roche Diagonastics Corp.) was treated at 37 ° C. for 1 hour, and the cells were washed with PBS. Subsequently, the cover slip was treated for 30 minutes in the converter-POD and washed again with PBS. Diaminobenzidine tetrahydrochloride (DAB) was incubated at 25 ° C. for 10 minutes and cells were observed under an optical microscope. Strong TUNEL staining was observed in the DNase treated group. TUNEL staining was weakly observed in the experimental control group and setlorellix treated group, whereas strong TUNEL staining was observed in the group treated with tryptorelix-1 (FIG. 5C). These results indicate that tryptellix-1 induces apoptosis of prostate cancer cells, which means that this apoptosis process is an important mechanism for cell death.

        As described above, GnRH-2 agonist tryptorelix-1 and its analogs specifically act on prostate cancer cell death and induce cancer cell death by apoptosis, and thus can be usefully used as a prostate treatment agent.

<110> NEUROGENEX CO., LTD. <120> USE OF THE GONADOTROPIN-RELEASING HORMONE II ANTAGONISTS AND ITS          ANALOGUES <130> IPM-29666 <160> 10 <170> KopatentIn 1.71 <210> 1 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Trptorelix-1 <220> <221> MOD_RES <222> (1) Ac-D-Nal (2) <220> <221> MOD_RES <222> (2) D-Phe (4Cl) <220> <221> MOD_RES <222> (3) <223> D-Pal (3) <220> <221> MOD_RES <222> (6) <223> D-Cit <220> <221> MOD_RES <222> (10) <223> D-Ala <400> 1 Xaa Phe Xaa Ser Tyr Xaa Trp Tyr Pro Ala   1 5 10 <210> 2 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Trptorelix-2 <220> <221> MOD_RES <222> (1) Ac-D-Nal (2) <220> <221> MOD_RES <222> (2) D-Phe (4Cl) <220> <221> MOD_RES <222> (3) <223> D-Pal (3) <220> <221> MOD_RES <222> (6) <223> D-Cit <220> <221> MOD_RES <222> (10) <223> D-Ala <400> 2 Xaa Phe Xaa Ser His Xaa Trp Tyr Pro Ala   1 5 10 <210> 3 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Trptorelix-3 <220> <221> MOD_RES <222> (1) Ac-D-Nal (2) <220> <221> MOD_RES <222> (2) D-Phe (4Cl) <220> <221> MOD_RES <222> (3) <223> D-Pal (3) <220> <221> MOD_RES <222> (6) <223> D-Cit <220> <221> MOD_RES <222> (10) <223> D-Ala <400> 3 Xaa Phe Xaa Ser Tyr Xaa Trp Leu Pro Ala   1 5 10 <210> 4 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Trptorelix-4 <220> <221> MOD_RES <222> (1) Ac-D-Nal (2) <220> <221> MOD_RES <222> (2) D-Phe (4Cl) <220> <221> MOD_RES <222> (3) <223> D-Pal (3) <220> <221> MOD_RES <222> (6) <223> D-Cit <220> <221> MOD_RES <222> (8) <223> D-Tyr <220> <221> MOD_RES <222> (10) <223> D-Ala <400> 4 Xaa Phe Xaa Ser Tyr Xaa Trp Tyr Pro Ala   1 5 10 <210> 5 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> N1 <220> <221> MOD_RES <222> (1) Ac-D-Nal (2) <220> <221> MOD_RES <222> (2) D-Phe (4Cl) <220> <221> MOD_RES <222> (3) <223> D-Pal (3) <220> <221> MOD_RES <222> (6) <223> D-Arg <220> <221> MOD_RES <222> (10) <223> D-Ala <400> 5 Xaa Phe Xaa Ser Tyr Arg Trp Tyr Pro Ala   1 5 10 <210> 6 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> N2 <220> <221> MOD_RES <222> (1) Ac-D-Nal (2) <220> <221> MOD_RES <222> (2) D-Phe (4Cl) <220> <221> MOD_RES <222> (3) <223> D-Pal (3) <220> <221> MOD_RES <222> (6) <223> D-Arg <400> 6 Xaa Phe Xaa Ser Tyr Arg Trp Tyr Pro Gly   1 5 10 <210> 7 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> N3 <220> <221> MOD_RES <222> (1) Ac-D-Nal (2) <220> <221> MOD_RES <222> (2) D-Phe (4Cl) <220> <221> MOD_RES <222> (3) <223> D-Pal (3) <220> <221> MOD_RES <222> (6) <223> D-Lys <220> <221> MOD_RES <222> (10) <223> D-Ala <400> 7 Xaa Phe Xaa Ser Tyr Lys Trp Tyr Pro Ala   1 5 10 <210> 8 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> N4 <220> <221> MOD_RES <222> (1) Ac-D-Nal (2) <220> <221> MOD_RES <222> (2) D-Phe (4Cl) <220> <221> MOD_RES <222> (3) <223> D-Pal (3) <220> <221> MOD_RES <222> (6) <223> D-Lys <400> 8 Xaa Phe Xaa Ser Tyr Lys Trp Tyr Pro Gly   1 5 10 <210> 9 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> N5 <220> <221> MOD_RES <222> (1) Ac-D-Nal (2) <220> <221> MOD_RES <222> (2) D-Phe (4Cl) <220> <221> MOD_RES <222> (3) <223> D-Pal (3) <220> <221> MOD_RES <222> (6) <223> D-Cit <400> 9 Xaa Phe Xaa Ser Tyr Xaa Trp Tyr Pro Gly   1 5 10 <210> 10 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Cet <220> <221> MOD_RES <222> (1) Ac-D-Nal (2) <220> <221> MOD_RES <222> (2) D-Phe (4Cl) <220> <221> MOD_RES <222> (3) <223> D-Pal (3) <220> <221> MOD_RES <222> (6) <223> D-Cit <220> <221> MOD_RES <222> (10) <223> D-Ala <400> 10 Xaa Phe Xaa Ser Tyr Xaa Leu Arg Pro Ala   1 5 10

Claims (6)

A gonadotropin releasing hormone II agonist, the peptide which induces apoptosis of prostate cancer cells, comprising the amino acid sequence of SEQ ID NO: 1 or an amino acid sequence having at least 70% homology therewith. The peptide of claim 1, which has an amino acid sequence of SEQ ID NO: 7 to SEQ ID NO: 9, inducing apoptosis of prostate cancer cells. A DNA or RNA oligonucleotide comprising a nucleotide sequence encoding the amino acid sequence of the peptide inducing apoptosis of the prostate cancer cell of claim 1. A pharmaceutical composition for treating prostate cancer containing the peptide of claim 1 or 2 as a pharmaceutical carrier and an active ingredient. A pharmaceutical composition for treating prostate cancer containing a gonadotropin releasing hormone II agonist as a pharmaceutical carrier and an active ingredient. Gonadotropin-releasing hormone II agonist for the treatment of prostate cancer.

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