TW201402614A - Zinc finger-like peptide, and pharmaceutical compositions containing the same for treating or preventing prostate cancer - Google Patents

Abstract

The present invention relates to a zinc finger-like peptide and a pharmaceutical composition containing the same for treating or preventing prostate cancer. The zinc finger-like peptide comprises at least seven amino acids, and the amino acid sequence of the at least seven amino acids has 85-100 similarity to a sequence represented by SEQ ID NO: 1. Besides, the present invention also relates to a pharmaceutical composition containing the same for treating and preventing prostate cancer.

Description

Treatment or prevention of zinc finger peptides such as prostate cancer, their plastids, and pharmaceutical compositions containing the same

The present invention relates to a zinc finger peptide which treats or prevents prostate cancer, and a pharmaceutical composition comprising the same.

The prostate (or prostate) is a gland belonging to the male reproductive system. If a genetic mutation in the cell causes the proliferation to be out of control, it can lead to prostate cancer. With the advancement of diagnostic techniques, the incidence of diagnosis of prostate cancer has increased year by year, and the number of deaths due to prostate cancer has increased by more than 40% in the past decade. According to statistics, the prostate cancer system is one of the top ten cancers, and prostate cancer is the second most common cancer in men, and the number of people killed is second only to lung cancer.

In general, when prostate cancer occurs, it will be like a benign prostate gland, and then it will be pressed into the urethra and there will be symptoms of urinary tract, such as frequent urination, drips, and delayed urine. In addition, when the prostate cancer becomes more serious, it may even invade the bladder or urethra and hematuria occurs. At the same time, in a small part of the tumor, the tumor may even be pressed to the ureter or urethra on both sides, leading to renal dysfunction, which evolves into uremia.

At present, the treatment of prostate cancer includes surgery, radiation therapy, hormone therapy, chemotherapy, and computerized knife radiosurgery. Among them, computerized dental surgery can greatly reduce side effects such as incontinence and sexual dysfunction, especially It can treat local non-metastatic prostate cancer, so It is an emerging treatment for prostate cancer. However, these methods still fail to achieve the efficacy of preventing prostate cancer.

Even though prostate cancer is more common in older men and the growth rate of prostate cancer is slower, early diagnosis and early treatment are still important guidelines for the protection of prostate cancer. Therefore, there is still a need to provide a pharmaceutical composition that can treat or even prevent prostate cancer, in order to reduce the incidence or mortality of prostate cancer.

The main object of the present invention is to provide a zinc finger peptide for treating or preventing prostate cancer and a pharmaceutical composition comprising the same, which can be treated by administering an effective dose of a zinc-like finger peptide to a desired individual. Prevent the purpose and efficacy of prostate cancer.

Further, another object of the present invention is to provide a zinc finger peptide for treating or preventing prostate cancer and a pharmaceutical composition comprising the same, which can express a type of zinc finger peptide in vivo to achieve therapeutic or prophylactic treatment. The purpose and efficacy of prostate cancer.

To achieve the above object, the present invention provides a zinc finger peptide for treating or preventing prostate cancer, comprising: at least seven amino acids having an amino acid sequence of 85-100% of the sequence of SEQ ID NO: 1. Similarity.

In addition, the present invention further provides a pharmaceutical composition for treating or preventing prostate cancer, comprising: an effective dose of a zinc-like finger peptide, wherein the zinc finger peptide comprises at least seven amino acids, and at least seven Amino acid The amino acid sequence has 85-100% similarity to the sequence of SEQ ID NO: 1; and a pharmaceutically acceptable carrier.

In the zinc finger peptide of the present invention and the pharmaceutical composition comprising the same, the zinc-like finger peptide refers to a small molecule protein similar to the zinc finger peptide, and the length of the amino acid sequence is not particularly limited, but may be It is composed of 7-200 amino acids, preferably 7-100 amino acids, more preferably 7-70 amino acids, still more preferably 7-45 amino acids, and even more preferably 7- 31 amino acids, and most preferably consist of 7-15 amino acids. Wherein, the zinc-like finger peptide has a structure similar to that of a zinc finger, optionally comprising a plurality of repeating amino acids of the same type, or may comprise a plurality of repeating identical amino acids, forming a bond-specific Protein motif (motif). The zinc finger peptide of the present invention can perform self-polymerization in an enzyme-free catalytic environment and form a good bond with several proteins in the cell, so that the zinc-like finger peptide can interact with the protein. The bond reaction further inhibits the growth of cancer cells and achieves the effect of treating or preventing prostate cancer.

Further, in the zinc finger peptide of the present invention and a pharmaceutical composition comprising the same, comprising at least seven amino acids, the amino acid sequence may have a 70-100% similarity to the sequence of SEQ ID NO: 1, Preferably, it has 85-100% similarity to the sequence of SEQ ID NO: 1. More preferably, the amino acid sequence of at least seven amino acids contained in the zinc-like finger peptide may have 70-100% identity with the sequence of SEQ ID NO: 1, preferably with SEQ ID NO: The 85-100% identity of the 1 sequence.

The zinc finger peptide of the present invention and the pharmaceutical composition comprising the same comprise at least seven 70-100% similarities to the sequence of SEQ ID NO: 1. Amino acid means that any short peptide comprising an amino acid sequence having 70-100% similarity to the sequence of SEQ ID NO: 1 may be a zinc finger peptide such as the present invention. For example, the amino acid sequence of such a zinc finger peptide can be an amino acid sequence having 70-100% similarity to the sequence of SEQ ID NO: 2, preferably having a sequence of 85 to SEQ ID NO: 100% similarity. More preferably, the amino acid sequence of the zinc-like finger peptide may have a 70-100% identity to the sequence of SEQ ID NO: 2, preferably 85-100% identity to the sequence of SEQ ID NO: 2.

Further, the sequence of the zinc finger peptide of the present invention may also have a 50-100% similarity to SEQ ID NO: 3, preferably 70-100% similarity, more preferably 80-100% similarity. In addition, the amino acid sequence of such a zinc finger peptide may be 50-100% identical to SEQ ID NO: 3, preferably 70-100% identical, more preferably 80-100% identical. Sex. Moreover, such a zinc finger peptide is SEQ ID NO: 3.

Further, the sequence of the zinc finger peptide of the present invention may have a 50-100% similarity to SEQ ID NO: 4, preferably 70-100% similarity, more preferably 80-100% similarity. . In addition, the amino acid sequence of such a zinc finger peptide may be 50-100% identical to SEQ ID NO: 4, preferably 70-100% identical, more preferably 80-100% identical. Sex. Furthermore, the amino acid sequence of such a zinc finger peptide is SEQ ID NO:4.

The zinc finger peptide amino acid sequence described above may be a zinc finger peptide sequence of any species, preferably a zinc finger amino acid sequence such as human or mouse. The zinc finger nucleotide sequence corresponding to the zinc-like finger peptide amino acid sequence can be obtained by genetic engineering artificial synthesis, or by human or mouse, the nucleotide expression sequence of the zinc-like finger peptide is isolated. In the present invention, The test was carried out using a zinc finger amino acid sequence such as a mouse, but the present invention is not limited thereto. For example, when treating a human, a human zinc finger amino acid sequence corresponding to a zinc finger amino acid sequence such as a mouse can be selected.

The method for obtaining the zinc finger peptide of the present invention is not limited to any form, and in addition to the performance of the protein by the plastid, the zinc finger peptide of the present invention can also be obtained by artificially synthesizing the peptide sequence.

In a specific embodiment of the zinc finger peptide of the present invention and a pharmaceutical composition comprising the same, the zinc-like finger-supplemented amino acid sequence is the sequence shown in SEQ ID NO: 2, or the amine of the zinc-like finger peptide. The acid sequence is the sequence shown in SEQ ID NO: 1, but the invention is not limited thereto.

In the present invention, the term "similarity" means the percentage of similar components, and except for the identical amino acid residues, amino acid residues having similar properties are assigned to similar amino acid residues. For example, amino acids containing saturated hydrocarbon groups such as Alanine, Valine, Leucine, and Isoleucine are assigned to similar amino acid residues; Aromatic groups containing amino acids such as Phenylalanine, Tyrosine, Tryptophan, and Histidine are assigned to similar amino acid residues; aspartic acid ( Aspartic acid), aspartic acid (Asparagine), glutamic acid (Glutamic acid), glutamine (Glutamine) and other amino acids containing additional acid groups (and their guanamines) are assigned to similar amine groups. Acid residues; amino acid containing additional amine groups such as Lysine and Arginine are assigned to similar amino acid residues; Serine and Threonine The amino acid containing an alcohol group is assigned to a similar amino acid residue; and the methyl amino acid (Methionine), cysteine A sulfur-containing amino acid such as (Cysteine) is assigned to a similar amino acid residue. In addition, the term "consistency" refers to the percentage of identical components, that is, must be identical amino acid residues.

The zinc finger peptide of the present invention comprises an amino acid sequence of at least seven amino acids, preferably an amino acid of the same type comprising at least two repeats, and the amino acid of the same type may be a hydrophilic amino acid. , such as: Aspartic acid, Glutamine, Serine, Threonine, or cysteine; or, preferably, at least two a repeating alcohol-based amino acid, such as: siminic acid, serine, tyrosine; or, preferably, at least two repeating thiol-type amino acids, such as : cysteine. Here, the zinc-like finger peptide preferably comprises at least two cysteine (Cycteine) and one histidine (Histidine) to form a zinc finger protein similar to the C2H2 type.

The present invention achieves the efficacy of treating or preventing prostate cancer by administering an effective dose of a zinc finger peptide to a desired individual. A pharmaceutically acceptable carrier can be included in the pharmaceutical composition of the present invention, and the carrier can be at least one selected from the group consisting of an active agent, an adjuvant, a dispersing agent, a wetting agent, and a suspending agent. Herein, the carrier to be included must be an acceptable carrier, and must be compatible with the zinc-like finger peptide in the pharmaceutical composition, preferably to stabilize the zinc-like finger peptide, and not to cause harm to the individual.

The term "treatment" refers to the administration of an effective dose of a zinc finger peptide to a desired individual to cure, alleviate, alleviate, affect, or improve the development or propensity of the disease. The term "prevention" refers to the administration of an effective dose of a zinc finger peptide to a desired individual to prevent, inhibit, or block the disease. Development or inclination. Herein, the individual in question generally refers to any individual who has a tendency to develop prostate cancer, has symptoms of prostate cancer, or has had an early, intermediate, and terminal stage of prostate cancer. The individual described above can be identified by a medical professional based on any suitable diagnostic method, such as: long-term exposure to the sun or exposure to an individual having a radiation environment, or an individual having a history of prostate cancer among family members.

The pharmaceutical composition provided by the present invention can be administered by non-oral, oral, nasal, topical, or via an implanted reservoir or the like. Wherein, oral refers to administration of a drug in capsules, lozenges, granules, sprays, syrups or other suitable forms; parenteral refers to subcutaneous injection, intradermal injection, intravenous injection, intramuscular injection, Intra-articular injection, intra-arterial injection, intra-articular injection, intrathoracic injection, intraspinal injection, intralesional injection, or intracranial injection, and any suitable infusion technique is administered.

In summary, the present invention provides a zinc-like finger peptide which can be a possible amino acid sequence of the above various arrangements. The pharmaceutical composition comprising the zinc-like finger peptide of the present invention can achieve the treatment or prevention of various prostate cancers by inhibiting the growth and formation of cancer cells.

Live experiments were performed using 8 week old mice. Each mouse was acclimatized for 2 weeks at room temperature and appropriate humidity before the experiment. Sterile water and standard meals are supplied daily during the adaptation period. After the mice were adapted, the zinc-like finger peptides and tumor cells were separately injected for observation.

Example 1

A zinc-like finger peptide synthesized by Genemed Synthesis Inc. (San Antonio, TX, USA), such as the zinc finger peptide shown in the sequence of SEQ ID NO: 1, is dissolved in degassed sterile water. In water), a fresh zinc-like finger peptide solution (100 μl) was prepared at a concentration of 2 mM. Herein, the zinc-like finger peptide is dissolved by using degassed sterile pure water to avoid the mass polymerization of the peptide and lose the original function of inhibiting the growth of cancer cells.

First, a zinc finger peptide solution (100 μl) configured as described above was injected into a nude mouse via a tail vein, and once a week, for 3 weeks.

Three months after the injection of the zinc-like finger peptide solution, two million cells of human prostate cancer cell line DU145 were inoculated on both sides of each mouse (ie, left and right), and the mice were continuously observed every day. Tumor volume size (mm ³ ). The results are shown in Figure 1A.

Example 2

The experimental procedure of this example was the same as in Example 1, except that the concentration of the zinc-like finger peptide solution was 4 mM.

Comparative example 1

The experimental procedure of this comparative example was the same as in Example 1, except that deionized water was used in place of the zinc-like finger peptide solution. The observation results are shown in Fig. 1B.

As shown in FIG. 1A, the experimental results of Example 1 show that when a mouse is first injected with a zinc-like finger peptide and then attacked by a prostate cancer cell, the cancer cell can be slowed down or even reduced in the early stage of cancer cell attack. Tumor volume. In addition, no tumor hyperplasia was observed after a period of continuous observation. Meanwhile, the experimental results of Example 2 were also similar to those of Example 1.

Accordingly, the results of Example 1 and Example 2 demonstrated that the pre-injection of the zinc-like finger peptide three months before the cancer cell attack can achieve the long-term prevention of prostate cancer.

However, as shown in FIG. 1B, when the zinc-like finger peptide is not injected into the mouse, when the prostate cancer cells are attacked, the tumor system implanted in the mouse is continuously proliferated on the left or right side of the mouse; Compared with the experimental results of Example 1 of Fig. 1A, the tumor of Comparative Example 1 was significantly larger.

Example 3

The experimental procedure of this example is the same as that of Example 1, except that after a continuous injection of a 3-week zinc-like finger peptide solution (100 μl) for one week, on the 20th and 90th days, two mice per mouse. The side (ie, left and right) was inoculated with two million cell numbers of human prostate cancer cell line PZ-HPV-7 (two million cell numbers). The observation results are shown in Fig. 2A.

Comparative example 2

The experimental procedure of this comparative example was the same as in Example 3 except that deionized water was used in place of the zinc-like finger peptide solution. The observation results are shown in Fig. 2B.

As shown in Fig. 2A, the experimental results of Example 3 showed that when the mouse was first injected with a zinc-like finger peptide and then attacked with prostate cancer cells, the proliferation of cancer cells was slowed down. In addition, even after multiple injections of cancer cells and continuous observation for a period of time, no tumor hyperplasia was observed.

However, as shown in Fig. 2B, when the zinc-like finger peptide was not injected before the mouse, when a large amount of prostate cancer cells were attacked, tumor proliferation was observed regardless of the left or right side of the mouse.

In summary, the zinc finger peptide of the present invention can not only treat prostate cancer, but also block the growth of prostate cancer cells and achieve the effect of preventing prostate cancer. Accordingly, the zinc finger peptide of the present invention can be used as an effective drug for treating or preventing prostate cancer.

The above-mentioned embodiments are merely examples for convenience of description, and the scope of the claims is intended to be limited to the above embodiments.

Fig. 1A is a graph showing the tumor volume of a mouse according to Example 1 of the present invention over time.

Fig. 1B is a graph showing the tumor volume of the mouse of Comparative Example 1 of the present invention over time.

Figure 2A is a graph showing tumor volume over time in a mouse of Example 3 of the present invention.

Fig. 2B is a graph showing the tumor volume curve of the mouse of Comparative Example 2 of the present invention over time.

<110> National Cheng Kung University National Cheng Kung University

<120> Zinc finger-like peptide, and pharmaceutical composition for treating or preventing prostate cancer

<130> S4335(100-079-TW2)

<160> 4

<170> PatentIn version 3.3

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Claims (13)

A zinc finger peptide for treating or preventing prostate cancer, comprising: at least seven amino acids having an amino acid sequence having 85-100% similarity to the sequence of SEQ ID NO: 1. A zinc finger peptide as described in claim 1, wherein the amino acid sequence of the at least seven amino acids has an 85-100% identity to the sequence of SEQ ID NO: 1. A zinc finger peptide as described in claim 1, wherein the amino acid sequence of the at least seven amino acids is SEQ ID NO: 1. A zinc finger peptide as described in claim 1, wherein the amino acid sequence of the zinc finger peptide is SEQ ID NO: 1. A zinc finger peptide as described in claim 1, wherein the amino acid sequence of the zinc finger peptide has 80-100% similarity to the sequence of SEQ ID NO: 2. A zinc finger peptide as described in claim 1, wherein the amino acid sequence of the zinc finger peptide is SEQ ID NO: 2. A pharmaceutical composition for treating or preventing prostate cancer, comprising: an effective dose of a zinc-like finger peptide, wherein the zinc finger peptide comprises at least seven amino acids, and the amine of the at least seven amino acids The acid sequence has 85-100% similarity to the sequence of SEQ ID NO: 1; and a pharmaceutically acceptable carrier. The pharmaceutical composition according to claim 7, wherein the amino acid sequence of the at least seven amino acids has an 85-100% identity with the sequence of SEQ ID NO: 1. The pharmaceutical composition according to claim 7, wherein the amino acid sequence of the at least seven amino acids is SEQ ID NO: 1. The pharmaceutical composition according to claim 7, wherein the amino acid sequence of the zinc finger peptide is SEQ ID NO: 1. The pharmaceutical composition according to claim 7, wherein the amino acid sequence of the zinc finger peptide has 80-100% similarity to the sequence of SEQ ID NO: 2. The pharmaceutical composition according to claim 7, wherein the amino acid sequence of the zinc finger peptide is SEQ ID NO: 2. The pharmaceutical composition according to claim 7, wherein the carrier is at least one selected from the group consisting of an active agent, an adjuvant, a dispersing agent, a wetting agent, and a suspending agent.