WO1998006424A1

WO1998006424A1 - Cancerous metastasis inhibitors for oral administration

Info

Publication number: WO1998006424A1
Application number: PCT/JP1997/002685
Authority: WO
Inventors: Hiroyuki Tsuda; Masaaki Iigo; Mamoru Tomita; Seiichi Shimamura; Zenta Takatsu; Kazunori Sekine
Original assignee: Morinaga Milk Industry Co., Ltd.
Priority date: 1996-08-15
Filing date: 1997-08-01
Publication date: 1998-02-19
Also published as: JPH1059864A

Abstract

Cancerous metastasis inhibitors for oral administration which contain as the active ingredient one or more substances selected from the group consisting of iron-free saturated lactoferrin, hydrolyzates of lactoferrins, pharmaceutically acceptable derivatives of these hydrolyzates, pharmaceutically acceptable salts of these hydrolyzates, peptides originating in the hydrolyzates of lactoferrins, pharmaceutically acceptable derivatives of these peptides, and pharmaceutically acceptable salts of these peptides. These cancerous metastasis inhibitors are reduced in side effects and can be orally administered over a long period of time, thus exerting inhibitory effects on cancerous metastasis.

Description

Technical blockage Oral cancer transfer control agent Technical field

The present invention relates to a cancer metastasis inhibitor that can be administered orally. More specifically, the present invention relates to a non-ferrous saturated lactoferrin, a hydrolyzate of lactoferrin, a pharmaceutically acceptable derivative of the hydrolyzate, a pharmaceutically acceptable salt of the hydrolyzate, and a hydrolysis of lactoferrin. Containing, as an active ingredient, one or two substances selected from the group consisting of a peptide derived from a product, a pharmaceutically acceptable derivative of the peptide, and a pharmaceutically acceptable salt of the peptide. It relates to a cancer metastasis inhibitor. Background art

Cancer diagnosis and treatment techniques have advanced remarkably, and the cure rate of cancers detected earlier has been improving. However, cancers that once seem to have healed often grow again in other organs, and the problem of metastasis suppression in cancer clinical practice is increasing in importance. The outline of metastasis is the release of cancer from the primary tumor, invasion into blood vessels or lymph nodes, migration in the bloodstream or lymph, adhesion to vascular endothelium, basement membrane or lymph nodes, and specific organs. It is known that it undergoes a number of processes, such as the propagation of germ (Experimental Medicine, Vol. 12, No. 8, pp. 906-911, 1994), and any of these processes, or By inhibiting all, metastasis of cancer cells can be suppressed.

Conventionally, many anticancer drugs have been used for the purpose of inhibiting metastasis, but their effect on inhibiting metastasis has not been clarified (Article of the General Meeting of the Cancer Society of Japan in 1994, p. 458, p. Year). In addition, many new substances have a major focus on suppressing cancer metastasis. Carbostiryl derivatives (JP-A-8-81443) which can suppress metastasis invasion as a metalloprotease inhibitor present in the extracellular matrix. Aminohalogenonaphthoquinone derivatives (JP-A-8-113555) which are known to prevent the adhesion of cancer cells are known.

Regardless of whether the drug is administered orally or parenterally, these various biological activity inhibitors act on cells other than cancer cells when administered in an effective amount. There are strong side effects, and it is not appropriate to administer an effective dose in terms of the quality of life (QOL) of patients. Therefore, a satisfactory cancer metastasis inhibitor, oral or parenteral, has not yet been put to practical use.

Cancer swelling often occurs early from the primary tumor, such as breast cancer, but metastasis recurs even after 5 and 10 years, as the survival rate after 5 years can be interpreted as a cure. Shigeru (Toshima Shigeru, Cancer Recurrence and Metastasis, p. 64, Jiyokuminsha, September 10, 1979, and Internal Medicine, Vol. 49, No. 6, pp. 1061-1066, 1982 ). Therefore, cancer patients who have been discharged from hospital usually use cancer metastasis inhibitors continuously at home medical treatment for several years to more than 10 years, while injections are used after home medical treatment or reintegration. Even if it is effective, it is necessary to go to the hospital for a long time even if it is effective, and it is a drug that is difficult to use practically for busy patients. Under such circumstances, there has been a long-awaited need for an orally-administered drug that can be easily used with ease and has few side effects.

Docosahexaenoic acid (DHA) and its derivatives (Japanese Unexamined Patent Publication No. Hei 8-53351) and perilla leaf extract (Japanese Unexamined Patent Publication No. Hei 8-73371), which are natural substances, are substances meeting such demands. Has been developed. These oral cancer metastasis inhibitors derived from natural products generally have few side effects, but are often ineffective, and have unique flavors and properties, and are liable to be oxidized. It is a substance having properties. Therefore, foods that have high personal taste and can be mixed are limited. For many patients, it was difficult to use the drug for a long time and continuously without resistance to control cancer metastasis.

On the other hand, lactoferrin is present in bodily fluids of mammals including humans, such as milk and saliva, tears, and mucous secretions, and has an approximately 10% sugar chain content and an iron-binding molecular weight of around 80,000.耱 As a protein, natural lactoferrin usually binds 10 to 20% of iron in a saturated state (hereinafter, non-iron-saturated lactoferrin may be referred to as Lf).

Lactoferrin is known to exhibit antibacterial activity against harmful microorganisms such as Escherichia coli, Candida and Clostridium [Journal of Pediatrics, Vol. 94, No. 1, 1979], also known to have antibacterial activity against staphylococci and enterococci [Journal of Dairy Science, 67, 606, 1984] ].

The present inventors have focused on the antibacterial properties of lactoferrin and have studied the acidity of mammalian lactoferrin, apolactoferrin, and metal-saturated or partially-saturated lactoferrin (hereinafter, these may be collectively referred to as Lf). Alternatively, it has been found that an enzymatic hydrolyzate has no undesirable side effects (for example, antigenicity) and has higher heat resistance and antibacterial properties than those of undegraded hydrolyzate. JP-A-5-320068, Kyoichi Noh << Patent Publication No. 438750>.

Further, the present inventors isolated peptides having strong antibacterial activity from hydrolyzates of Lf compounds, synthesized peptides having the same amino acid sequence as those peptides and derivatives of those peptides, An antimicrobial peptide consisting of 20 amino acid residues (JP-A-5-92994); an antibacterial peptide consisting of 11 amino acid residues (JP-A-5-78392); An antimicrobial peptide consisting of amino acid residues (JP-A-5-148297), an antibacterial peptide consisting of five amino acid residues (JP-A-5-1498296), and 3 to 6 amino acid residues Antibacterial properties consisting of base Invented a peptide (Japanese Patent Application Laid-Open No. 5-148295) and already filed a patent application. Furthermore, the present inventors added a peptide having the same amino acid sequence as the hydrolyzate of Lf or a derivative of these peptides to the brain. Protective action (JP-A-6-172200), action of promoting growth of fibroblasts by epidermal growth factor (JP-A-6-48955), and action of promoting nerve growth factor production (JP-A-5-23557) And each has already filed a patent application.

As an example of using lactoferrin as a therapeutic agent for diseases, an antirheumatic agent (JP-A-5-186368) is known, and lactoferrin hydrolyzate includes tyrosinase activity inhibition (European Patent Publication No. 438750), Prevention of adhesion of pathogenic bacteria to cells (JP-A-3-220130), antiviral action (JP-A-12332326) and the like are known.

The use of lactoferrin as an anticancer agent has also been studied. For example, iron-saturated lactof:!: Phosphorus is known to have an antitumor effect (Japanese Patent Publication No. 5-86932). The present inventors have discovered that a peptide having the same amino acid sequence as a substance obtained by hydrolyzing lactoferrin with an acid or an enzyme or a derivative of these peptides has a parenteral antitumor effect, and filed a patent application (Japanese Patent Application Laid-Open No. 7-1995). No. 309771).

Several studies have also been conducted on the suppression of cancer metastasis.Lactoferrin has anti-cancer metastasis activity by parenteral administration, and the results of studies indicate that lactoferrin is injected intravenously into tumor cells. Has been reported under the unique condition of intraperitoneal administration [Cancer's Research (Cancer Research), Vol. 54, pp. 2310-2312, 1994]. Furthermore, it has been reported that lactoferrin and a peptide consisting of specific 25 amino acids in the amino acid sequence of lactoferrin have a cancer metastasis inhibitory activity by parenteral administration (Agricultural Chemistry Society, Vol. 69, extraordinary). No. pp. 143, 1995). Thus, despite some studies, whether lactoferrin, lactoferrin hydrolyzate, or a lactofer:!: Phosphorus-derived peptide with a specific amino acid sequence, etc., has cancer metastasis inhibitory activity by oral administration. This has not been studied in the past. The reason is that when lactoferrin is orally administered, it is easily degraded by the action of an enzyme, and therefore cannot exhibit a high level of physiological activity, especially its activity in suppressing cancer metastasis. This is due to the common sense of protein chemistry.

Furthermore, lactoferrin hydrolyzate and peptides derived from lactofer:!: Phosphorus are more likely to be degraded by digestive enzymes than proteins before hydrolysis. It was not being considered.

In view of the above, an oral cancer metastasis inhibitor which can be used for a long period of time and has few side effects has been desired, but it is a reality that no excellent substance is known yet, and lactoferrin, a hydrolyzate of lactoferrin However, it has not been reported that lactoferrin-derived peptide has a cancer metastasis inhibitory effect by oral administration, and it is not described in the literature.

The present invention has been made in view of the above prior art, and has as its object to provide a cancer metastasis inhibitor that can be administered for a long time with few side effects and that can be administered orally. Disclosure of the invention

The present inventors have searched for a cancer metastasis inhibitor that has few side effects, can be administered for a long period of time, and can be administered orally, but surprisingly, digestion by digestive enzymes is inevitable. Lf has been found to have a cancer metastasis inhibitory effect by oral administration. In the process of studying this, a more surprising phenomenon was the hydrolysis of lactoferrins, which are more susceptible to degradation by digestive enzymes than Lf. In addition, the inventors have found that a peptide derived from lactoferrins has an effect of suppressing cancer metastasis by oral administration, and repeated tests for confirming the effectiveness thereof, thereby completing the present invention.

The present invention relates to non-iron-saturated lactofu:!: Phosphorus, a hydrolyzate of lactoferrin, a pharmaceutically acceptable derivative of the hydrolyzate, a pharmaceutically acceptable salt of the hydrolyzate, and a lactoferrin. Contains, as an active ingredient, one or two substances selected from the group consisting of hydrolyzate-derived peptides, pharmaceutically acceptable derivatives of the peptides, and pharmaceutically acceptable salts of the peptides. To provide an oral cancer metastasis inhibitor.

The oral cancer transfer inhibitor of the present invention includes non-iron-saturated lactofurin, a hydrolyzate of lactoferrins, a pharmaceutically acceptable derivative of the hydrolyzate, or a pharmaceutically acceptable salt of the hydrolyzate. In a preferred embodiment, the compound is orally administered at a rate of 3 kg / kg of body weight.

Further, the oral cancer transfer inhibitor of the present invention may be a peptide derived from a hydrolyzate of lactoferrin, a pharmaceutically acceptable derivative of the peptide, or a pharmaceutically acceptable salt of the peptide. It is a desirable embodiment that the compound is orally administered at a rate of 2 to 3 20 mg / kg body weight.

Further, in the oral cancer metastasis inhibitor of the present invention, it is preferable that the peptides have the amino acid sequence of any one of SEQ ID NO: 1 to SEQ ID NO: 31.

Further, the present invention also provides a food composition and a feed composition containing the above-mentioned oral cancer transfer inhibitor as an essential component. BEST MODE FOR CARRYING OUT THE INVENTION

Lf, which is one of the active ingredients of the oral cancer metastasis inhibitor of the present invention, is a commercially available product or is isolated from mammalian milk by a conventional method, and is isolated from milk because of its large production amount. Things are desirable. An example of the separation and purification of Lf is as follows. CM-Sepharose FF (Pharmacia) was packed in a column, hydrochloric acid was passed through, washed with water, the ion exchanger was equilibrated, and skim milk with a pH of 6.9 cooled to 4 ° C was passed through the column. Then, the permeate is collected and passed through the column again. Next, distilled water is passed through the column, and saline is passed through to obtain an eluate of the basic protein adsorbed on the ion exchanger.

To this eluate, ammonium sulfate was added at a saturation of 80% to precipitate the protein, and the precipitate was collected by centrifugation, washed with an ammonium sulfate solution having a saturation of 80%, and deionized water was removed. The resulting solution was ultrafiltered using an ultrafiltration membrane module (for example, SLP 005 3 manufactured by Asahi Kasei Corporation), water was added, and then filtration was performed using the same apparatus. , Desalting and freeze-drying to obtain powdered lactolactoferrin.

The purity of Lf obtained by the above method was measured by electrophoresis, and as a result, it was found to have a purity of 95% or more (weight; hereinafter the same, except for the decomposition rate, unless otherwise specified). It is needless to say that the lactofurin-containing solution in each purification step before freeze-drying can be used in the present invention.

Although human Lf cannot be produced in large quantities, it is human Lf obtained from recombinant fungi or dairy cows (transgenic 'caws) obtained by recombinant DNA technology. Can also be used in the present invention.

The effective dose of Lf is in the range of 3 to 3200 mg / kg body weight per day based on the results of animal studies.

When lactoferrin hydrolyzate or peptide, which is another active ingredient of the oral cancer metastasis inhibitor of the present invention, is produced from lactoferrin, lactoferrin used as a starting material is commercially available lactoferrin, mammals (for example, human, Colostrum, sheep, goats, goats, etc., colostrum, transitional milk, normal milk, end-stage milk, etc., or the processed products of these milks, such as skim milk, whey, etc., in a conventional manner (eg, ion exchange) Lf separated by chromatography, etc., and Apolactov which was deferred with hydrochloric acid, citric acid, etc .:! : Lin, a It is a metal-saturated or partially-saturated lactoferrin obtained by chelating polactopherin with a metal such as iron, copper, zinc, and manganese, and a commercially available product or a preparation produced by a known method can also be used.

The decomposed product of Lf can be obtained by hydrolyzing Lf using an acid or an enzyme. Hydrolysis with an acid is performed by dissolving Lf in a concentration of 0.1 to 20%, preferably 5 to 15%; water or purified water; and adding an inorganic acid such as hydrochloric acid or phosphoric acid, or quinic acid to the resulting solution. The pH of the solution is adjusted to 1 to 4, preferably 2 to 3, and the mixture is heated at an appropriate temperature for a predetermined time according to the adjusted pH to hydrolyze. For example, when adjusted to pH 1-2, 80-130 ° C, desirably <90-120 ° 〇, and adjusted to pH 2-4, 100-130 ° C, desirably 100-120 ° (: And, respectively, heating for 1 to 120 minutes, preferably 5 to 60 minutes.

Enzymatic hydrolysis involves dissolving Lf in water or purified water at a concentration of 0.5 to 20%, preferably 5 to 15%, adjusting the pH of the solution to the optimal pH of the enzyme to be used, and ° C, preferably 30 to 50 ° C, and hold and hydrolyze for 30 to 600 minutes, preferably 60 to 300 minutes. Deactivates enzymes.

There are no particular restrictions on the enzymes used. Commercially available enzymes, for example, Morcine (trademark, manufactured by Morishin Pharmaceutical Co., Ltd., optimal pH 2.5-3.0), butapepsin (Wako Pure Chemical Industries, Ltd., optimal pH 2-3) , Sumiteam AP (trademark, manufactured by Shin Nippon Chemical Co., Ltd., optimal pH 3.0), Amano M (trademark, manufactured by Amano Corporation, optimal pH 7.0), tribcine (Novo Corp., optimal pH 8.0), etc. Used alone or optionally in combination.

The amount of enzyme used is in the range from 0.1 to 5.0%, particularly preferably from 0.5 to 3.0%, based on the substrate.

The rate of hydrolysis degradation is 4 to 50%, preferably 6 to 40%, as a percentage measured by the following method. In other words, this decomposition rate is the formol state measured by formol titration with respect to the total amount of nitrogen measured by the Kjeldahl method. Formula from nitrogen content

Decomposition rate (%) = (formol nitrogen content total nitrogen content) x 100

Is a value calculated by

The effective dose of Lf hydrolyzate is in the range of 3 to 320 Omg / kg of body weight per day, based on the results of animal studies.

The peptide derived from Lf, which is another active ingredient of the cancer metastasis inhibitor of the present invention, is the same as the peptide isolated from the hydrolyzate of the Lf by a known separation means. A peptide having the same or homologous amino acid sequence, a pharmaceutically acceptable derivative of these peptides, a pharmaceutically acceptable salt of these peptides, an amino acid sequence identical or homologous to these peptides It is a chemically synthesized peptide, or an arbitrary mixture thereof (hereinafter, these may be referred to as peptides).

For isolation of the peptide, for example, the filtrate of the hydrolyzate of Lf is neutralized with a sodium hydroxide solution, heated at 80 ° C for 10 minutes to inactivate the enzyme, cooled to room temperature, and centrifuged. Obtain a clear supernatant. The supernatant was subjected to reversed-phase high-performance liquid chromatography and eluted with a gradient of 20 to 60% acetonitrile containing 0.05 WTFA (trifluoroacetic acid), and fractions having an acetonitrile content of 27 to 30% were separated. The fraction is dried under vacuum to obtain peptides derived from Lf.

Alternatively, an automatic peptide synthesizer (Pharmacia LKB Biotechnology Inc.

LKBBiolynx4170) and solid-phase peptide synthesis by Shepard et al. [Jarnal of Chemical'Society. Perkin's Transaction 1: Organic and Bio-Organic Chemistry 8: 538, 1981 [Journal of Chemical Society. Peptides can also be synthesized as follows.

Amino acid whose amine function is protected by 9 fluorenylmethoxycarbonyl group [Hereinafter, sometimes referred to as Fmoc-amino acid or Fmoc-unique amino acid name (for example, Fmoc-asparagine)], N, N-dicyclohexylcarboximide is added to form a desired amino acid anhydride. This Fmoc-amino acid anhydride is used for the synthesis. To produce peptide chains, Fmoc-amino acid anhydride corresponding to the amino acid residue at the C-terminus is immobilized via its carboxyl group on Ultrocin A resin (Pharmacia LKB Biotechnology, Inc.) using dimethylaminopyridine as a catalyst. I do. The resin is then washed with dimethylformamide containing piperidine to remove the protecting group of the amine function of the C-terminal amino acid. Subsequently, the Fmoc-amino acid anhydride corresponding to the second from the C-terminal of the amino acid sequence is coupled to the deprotected amine functional group of the amino acid fixed to the resin via the C-terminal amino acid residue. Hereinafter, amino acids are sequentially fixed in the same manner. After coupling of all amino acids is completed and a peptide chain of the desired amino acid sequence is formed, removal of protecting groups and removal of the peptide with a solvent consisting of 94% TFA, 5% phenol, and 1% ethanediol. The peptide is purified by high performance liquid chromatography, and the solution is compressed and dried to obtain peptides derived from Lf by synthesis.

Furthermore, peptides having the same amino acid sequence or homologous amino acid sequence as these peptides, derivatives of these peptides, pharmaceutically acceptable salts of these peptides, or any mixture thereof are, for example, The methods described in the inventions of JP-A-5-92994, JP-A-5-78392, JP-A-5-148297, JP-A-5-1498296 and JP-A-5-148295 are disclosed. Can be obtained by

The effective dose of peptides derived from Lf is in the range of 0.2 to 320 mgZ / kg of body weight per day when integrated from the results of animal tests.

As the peptides obtained by the above method, peptides having the following amino acid sequence, derivatives or salts thereof can be exemplified as desirable embodiments. You. For example, peptides having the amino acid sequences of SEQ ID NOs: 1, 2, and 27, salts or derivatives thereof (JP-A-5-78392), peptides having the amino acid sequences of SEQ ID NOs: 3, 4, 5, and 6, and salts thereof Or a derivative thereof (JP-A-5-148297); a peptide having the amino acid sequence of SEQ ID NO: 8, 9, 9 or 31; a salt thereof or a derivative thereof (JP-A-5-1498296); Peptides having an amino acid sequence of 21; salts or derivatives thereof (JP-A-5-148295); Peptides having an amino acid sequence of amino acids 22 to 26, 28, 29, and 30; salts thereof; A derivative (JP-A-5-92994).

Examples of the pharmaceutically acceptable salts of the peptides include acid addition salts such as hydrochloride, phosphate, sulfate, citrate, lactate and tartrate. Amidated or acylated derivatives can be exemplified.

Lf, Lf hydrolysates, and Lf-derived peptides obtained as described above are formulated into oral dosage forms such as sugar-coated tablets, tablets, capsules, and nutrients by conventional methods, and formulated as enteral dosage forms. can do. Furthermore, it can be provided as a food composition by being blended with food such as a beverage or jelly, or a feed composition by being blended with feed.

Next, the present invention will be described in more detail with reference to test examples. Test example 1

This test was carried out to examine the effect of Lf and peptides derived from Lfs on oral cancer metastasis.

1: Test sample, test animal and tumor cell line

(a) Test sample

A commercially available Lf powder (manufactured by Morinaga Milk Industry Co., Ltd.) and the same method as in Reference Example 2 from this Lf powder Lf-derived peptide prepared by the above was used.

(b) Test animals

Ninety-six 6-week-old CDF1 mice (purchased from Nippon Charles Riva Co., Ltd.) were randomly used in 9 groups (10 mice per group).

(c) tumor cells

The method of Fiddler et al. [Nature, Vol. 242, pp. 148-149, 1973] was partially modified and prepared as follows. Mouse colon cancer colon 26 cells (hereinafter sometimes referred to as Co26 cells. Obtained from Cancer Research Institute, Cancer Institute, Cancer Chemotherapy Center) IX 10 ⁵ mice, 6-week-old Balb / c mice (Japan (Purchased from Charlriver Co., Ltd.) Three mice were injected into the tail vein, and about 3 weeks later, the formed lung metastasis tumor was removed and crushed, and the same number of released cells was injected again into the tail vein of three mice of the same strain I do. This was repeated several times to select a highly metastatic cell line, Co26Lu, which spontaneously metastasized to the lung, and was used in this study. In addition, it was found that Co26Lu metastasized about 14 after subcutaneous transplantation.

2: Test method

1 × 10 ⁵ Co26Lu cells were implanted subcutaneously on the back of all 90 CDF1 mice in 9 groups (day 0). Mice in each group were orally administered on day 5-9, 12-16 and 19-21 up to 300-1000 mg Lf of kg body weight or 30-100 mg Lf-derived peptide of kg body weight 1 time However, nothing was administered to the control group.

On the 22nd day, the lungs were excised, fixed with acetone, and the number of lung metastases was visually counted to test the effect of suppressing metastasis to the lungs.

One group in the Lf administration group received 150 mgZ / kg body weight twice daily (shown as 150 X 2 in Table 1), and one group in the peptide administration group received 15 mgZ day / kg body weight twice daily (Table 1). 15 X 2 in Table 1). 3: Test results

The results of this test are shown in Table 1. As is clear from Table 1, in the group to which Lf and the peptide derived from Lf were administered, the number of metastases almost decreased by half according to the dose, and the cancer metastasis inhibitory effect of these substances was clearly observed. More specifically, even with a dose of 300 mg / kg body weight per day in the Lf-administered group, it is more effective to divide the group twice a day and administer it twice a day. There was no significance in administering divided doses.

In addition, although the size of the transplanted tumor was reduced due to its toxicity with general metastasis inhibitors, in this test the size of the transplanted tumor was different between the control group and the Lf and Lf-derived peptide-administered groups. Therefore, their toxicity was estimated to be low. Furthermore, no metastasis to other organs such as the liver was observed by microscopic observation of each organ.

From the above results, it was clear that Lf and Lf-derived peptides had an effect of suppressing oral cancer metastasis. Similar tests were performed for other peptides, but almost the same results were obtained.

table 1

Amount transferred Number of transfers

Test group

(mg / day / kg body weight)

Control group 0 56.0 47- 95

1000 28.0 20-44

300 37.0 5-53

Lf group 100 44.5 10-67

150 X 2 27.5 21-111

100 31.0 6-49

Peptide 30 36.5 12-86

Group 10 47.5 35-71

15 X 2 38.0 21-65 Test example 2

This test was conducted to examine the effect of Lf, Lf hydrolyzate, and peptides derived from Lf compounds on oral cancer metastasis.

1: Test sample, test animal and tumor cell line

(a) Test sample

A commercially available Lf powder (manufactured by Morinaga Milk Products Co., Ltd. administered to Group 1), an Lf hydrolyzate prepared from this Lf powder by the same method as in Reference Example 1 (administered to Group 2), the same as Reference Example 2 Lf-derived peptide prepared from this hydrolyzate by the method (administered to Group 3) and Lf-derived peptide chemically synthesized by the same method as in Reference Example 3 (organic synthetic peptide; administered to Group 4) used. As a control, serum serum albumin (fraction V, manufactured by Sigma) was administered to the control group.

(b) Test animals

Twenty 6-week-old CDF1 mice (purchased from Nippon Charls Riva Co., Ltd.) were randomly used in 4 groups (5 mice per group).

(c) tumor cells

The highly metastatic lung cell line Co26Lu prepared in Test Example 1 was used.

2: Test method

After transplantation of Co26Lu cells, each group was treated in the same manner as in Test Example 1 except that administration was performed once daily from No. 5 to Day 27 at the dose shown in Table 2, and lungs were removed on Day 28. Tested by the same method.

3: Test results

The results of this test are shown in Table 2. As is evident from Table 2, the number of metastases was lower in all of the first to fourth groups than in the control group, and the effects of these administrations on suppressing cancer metastasis were observed. A similar test was performed for other Lf, Lf hydrolysates, and Lf-derived peptides, but almost the same results were obtained. Table 2

Test example 3

This test was performed to determine the acute toxicity of Lf hydrolysates.

1: Animal used

Twenty bisexual CD (SD) rats (purchased from Japan SLC) of 6 weeks of age were used, and male and female were randomly divided into four groups (five rats per group).

2: Test method

The Lf hydrolyzate produced by the same method as in Reference Example 1 was dissolved in water for injection (Otsuka Pharmaceutical Co., Ltd.), and a single gavage was administered at a rate of 4 ml Zl00 g body weight using a metal ball-pointed needle. Toxicity was tested. Dosage 1000, 2000 and 4000

1718 1 <£ body.

3: Test results

The results of this test are shown in Table 3. As is evident from Table 3, no deaths were observed in the groups that received this hydrolyzate at the rate of 1 OOOmgZkg and 2000mgZkg. Therefore, the LD _{5 of} this hydrolyzate. Was 200 OmgZkg body weight or more, and the toxicity was found to be extremely low. The same test was conducted for other hydrolyzates of Lf and peptides derived from Lf, but almost the same results were obtained. Table 3

Number of deaths

Amount

(mg / kg) male

0 0/5 0/5

1000 0/5 0/5

2000 0/5 0/5

4000 4 1 5 3/5 Test example 4

This test was performed to determine the acute toxicity of the peptides.

1.Use animal

Six-week-old CD (SD) rats (purchased from Japan SLC) of 35 amphoteric sex were used, and male and female were randomly divided into seven groups (five in each group).

2: Test method

An Lf-derived peptide (peptide purified from a hydrolyzate) produced by the same method as in Reference Example 2 and a peptide (organic synthesis peptide) produced by the same method as in Reference Example 3 were combined with water for injection (Otsuka Pharmaceutical Co., Ltd.). And a single gavage administration using a metal ball needle at a rate of 4 ml / 100 g body weight was conducted to test acute toxicity. The doses were 1000, 2000 and 4000 mgZ kg body weight.

3: Test results

The results of this test are shown in Table 4. As is evident from Table 4, there were no deaths in the groups administered at 1000 mg / kg body weight and 2000 mg / kg body weight, regardless of the peptide administered. Therefore, the LD50 of these peptides was 2000 mgZkg or more, indicating that their toxicity was extremely low. The same test was conducted for other peptides, but almost the same results were obtained. Table 4

Next, the present invention will be described more specifically with reference to examples. However, the present invention is not limited to the following examples. The Lf hydrolyzate and peptides used in each Example were prepared in the same manner as in the following Reference Examples. Reference Example 1: Preparation of Lf hydrolyzate

The hydrolyzate was prepared by using the following method using Lf.

Dissolve 500 g of commercial Lf (manufactured by Mirai) in 9.5 liters of purified water, add hydrochloric acid to the resulting solution to adjust the pH to 3.0, and then use commercially available pork pepper. (Manufactured by Wako Pure Chemical Industries, Ltd.) was added thereto and hydrolyzed at 37 ° C for 6 hours. Then adjust the pH to 7.0 with 6N sodium hydroxide, heat at 80 ° C for 10 minutes to inactivate the enzyme, cool to room temperature, filter through celite, freeze-dry the filtrate, powder About 470 _g of Lf hydrolyzate was obtained. The decomposition rate of the obtained hydrolyzate was measured by the same method as described above, and was found to be 30%. Reference Example 2: Preparation of peptides

Dissolve 50 mg of commercially available ゥ Lf (manufactured by Sigma) in 0.9 ml of purified water, adjust the pH to 2.5 with 0.1N hydrochloric acid, and then use commercially available butapepsin (manufactured by Sigma). 1 mg was added and hydrolyzed at 37 ° C for 6 hours. Then adjust the pH to 0.1 with 0.1N sodium hydroxide, inactivate the enzyme by heating at 80 ° C for 10 minutes, cool to room temperature, centrifuge at 15, OOOrpm for 30 minutes, and clear The supernatant was obtained. The supernatant (1 OO ju I) was subjected to high-performance liquid chromatography using TSK gel ODS- "! 20T (manufactured by Tosoichi Co., Ltd.) at a flow rate of 0.8 mlZ. Elute with 20% acetonitrile containing TFA (trifluoroacetic acid), then elute with a gradient of 20-60% acetonitrile containing 0.055 TFA for 30 minutes, and collect fractions eluted between 24 and 25 minutes The dried product was dissolved in purified water at a concentration of 2% (WV) and subjected to high performance liquid chromatography using TSK gel ODS-120T (manufactured by Tosoh I) to obtain 0.8 ml Eluted with 24% acetonitrile containing 05% TFA for 10 minutes after sample injection at a flow rate of 35.5 The fractions eluted during 5 minutes were collected and dried in vacuo to give the peptide.

The above peptide was hydrolyzed with 6N hydrochloric acid, and the amino acid composition was analyzed by an ordinary method using an amino acid analyzer. The same sample was subjected to Edman degradation 25 times using a gas-phase seek sensor (Applied Biosystems) to determine the sequence of 25 amino acid residues. In addition, a disulfide bond analysis method using DTNB [5,5-dicyclohexyl bis (2-dibenzobenzene) '] [Analytical Biochemistry], Vol. 67, No. 493 P. 1975] confirmed the presence of disulfide bonds.

As a result, this peptide consists of 25 amino acid residues, the 3rd and 20th cysteine residues form a disulfide bond, and 2 amino acid residues N-terminal from the 3rd cis-in residue. Group is located at the C-terminal side from the 20th cysteine residue It was confirmed that each of the five amino acids had the amino acid sequence of SEQ ID NO: 26 linked thereto. Reference Example 3: Organic synthesis of peptides

Solid-phase peptide synthesis method using Shepard et al. Using an automatic peptide synthesizer (Pharmacia LKB Biotechnology, Inc., LKB Biolynx4170) [Journal 'Saibu Chemical' Society. Perkin 'Transaction 1: Perpetual Nick-Andy Nozio Perpetual Nick' Chemistry (Journal of Chemical Society. Perkin ransaction 1: Organic and Bio-Organic Chemistry, p. 538, 1981) Was used to synthesize the peptide as follows.

N, N-Dicyclohexylcarbodiimide is added to an amino acid having its amine functional group protected with a 9-full-year-old enylmethoxycarbonyl group to form an anhydride of the desired amino acid, and this Fmoc-amino acid anhydride is synthesized. Using. In order to produce a peptide chain, Fmoc-asparagine anhydride corresponding to the C-terminal asparagine residue is converted to pertrosin A resin (Pharmacia LKB Biotechnology Co., Ltd.) via its carboxyl group using dimethylamino pyridine as a catalyst. Made). The resin is then washed with dimethylformamide containing piperidine to remove the protecting group for the amine function of the C-terminal amino acid. The Fmoc-arginine (Pmc: 2,2,5,7,8-pentamethy chromato-6-sulphonyl group) anhydride corresponding to the second from the C-terminal of the amino acid sequence is then replaced with the C-terminal amino acid residue Via the above, was coupled to the deprotected amine functional group of asparagine fixed to the resin. Glutamine, tributofan, glutamine, and phenylalanine were sequentially fixed in the same manner as described below. After coupling of all amino acids is completed and a peptide chain with the desired amino acid sequence is formed, removal of protecting groups and removal of peptide with a solvent consisting of 94% TFA, 5% phenol, and 1% ethanediol And purify the peptide by high performance liquid chromatography, concentrate this solution, dry it, A powder was obtained.

The peptide was analyzed for amino acid composition by a conventional method using an amino acid analyzer, and the sequence was determined using the same sequencer as in Reference Example 2.As a result, the peptide had the amino acid sequence of SEQ ID NO: 10. It was confirmed. Reference Example 4: Preparation of Apolactoferrin

Dissolve 1 kg of commercial L-powder powder (manufactured by Domo) prepared from skim milk in 20 liters of purified water, place in a dialysis tube (manufactured by Sanko Junyaku Co., Ltd. 1-7Z8), and add 400 liters of 0.1 Dialysis at 4 ° C for 36 hours against M citrate solution (pH 2.2) and then against 400 liters of deionized water at 4 ° C for 24 hours to remove citrate (2 times) The deionized water was exchanged), and the dialysis solution was centrifuged to remove solids and freeze-dried to obtain about 950 g of powdery apolactopherin. The saturation of iron was measured using the spectrophotometer (2000U, manufactured by Hitachi, Ltd.) for the obtained apolactoferin based on the absorbance at 450 nm. As a result, the saturation was 0%. Reference Example 5: Apolactov ;! : Preparation of phosphorus hydrolyzate

Using disiapolactoferin of Reference Example 4, its hydrolyzate was prepared by the following method. Was dissolved apo Lactobacillus Hue phosphorus 500g of purified water 9.5 liters, and adjust the _P H to 2.0 by addition of 1 M hydrochloric acid to give al solution was heated for 15 minutes at 120 ° C, cooled Thus, a hydrolyzate was obtained. Next, adjust the pH to 6 with 6M sodium hydroxide, filter through celite, freeze-dry the rapa solution, and powder Apolactov :! : About 400 g of phosphorus hydrolyzate was obtained. The degree of decomposition of this hydrolyzate was 9%. Reference Example 6: Preparation of Apolactoferrin-Derived Peptides

50 mg of the hydrolyzate of asiapolactoferrin from Reference Example 5 was dissolved in 0.9 rrH of purified water, and purified by the same method as in Reference Example 2 below. TSK gel 0D at the end of purification In the high performance liquid chromatography using S-120T (manufactured by Tosoichi Co., Ltd.), the fractions eluted between 35.5 and 37.5 minutes were collected and dried in vacuo to obtain the peptide derived from Apolactoferrin. Was.

The peptide was hydrolyzed in the same manner as in Reference Example 2, the amino acid composition was determined, the amino acid sequence was determined, and the presence of disulfide bonds was confirmed by disulfide bond analysis using DTNB.

As a result, this peptide consists of 32 amino acid residues, the 10th and 27th cysteine residues form a disulfide bond, and the 9th amino acid residue N-terminal from the 10th cyspine residue. It was confirmed that the group had the amino acid sequence of SEQ ID NO: 29 in which 5 amino acids were bonded to the C-terminal from the cysteine residue at position 27, respectively. Example 1: Preparation of powdered animal feed containing Lf

60 g of Lf (made by Mirai) and 3 kg of MF powder feed (manufactured by Oriental Yeast) are mixed for 30 minutes with a powder mixer (manufactured by Kanto Blender Co., Ltd., SS model, Model No. 151), and 500 g of polyethylene is mixed. Into 2% Lf-containing feed. This feed was stored in a refrigerator. Example 2: Preparation of enteral nutritional supplement containing Lf

Add 100 ml of hot water at 50-60 ° C to 2 g of powdered Lf (made by Mirai) and 1 g (89 g) of a commercially available enteral nutritional supplement (made by Eisai Co., Ltd. Clinimir) and mix evenly with a whisk. Then, 240 ml of hot water was further added to dissolve completely, and this was divided into several doses per day and administered. Example 3: Preparation of beverage containing Lf

90 g of skim milk powder (Morinaga Milk Industry Co., Ltd.) is dissolved in 800 ml of hot water at 50 ° C. (Nissin Seiko Co., Ltd.) 30 g, instant coffee powder (Nestlé Co., Ltd.) 14 g, caramel (Showa Kako Co., Ltd.) 2 g and coffee flavor (San-Ei Kagaku Co., Ltd.) 0.01 g were sequentially added with stirring and dissolved. After cooling to 10 ° C., a solution prepared by dissolving 1 g of Lf (Morinaga Dairy) in 75 ml of water was added to prepare a milk drink containing 1% of LfO. Example 4: Preparation of Lf powder

In a mortar, mix 10 g of commercial Lf powder (produced by Mirai) and 50 g of lactose (produced by Morinaga Milk Co., Ltd.) in a mortar. Add 40 g of lactose separated by Inuchi Seieido Co., Ltd. and mix. The whole amount is again separated with a No. 5 sieve, and 5 g per package is packed in a packing machine (Tokyo Shokai; OMP-90A), 10%. Lf powder was obtained. This powder was stored in a cool and dark place. Example 5: Preparation of rare cheese cake containing Lf

(1) Biscuits (made by Morinaga Seika Co., Ltd.) A total of about 160 g was shredded, mixed with 30 g of heated and melted butter (made by Morinaga Milk Products).

(2) The biscuit butter mixture (1) was spread in a round bottom container of 18 cm and pressed hard for $$.

(3) 5 g of powdered gelatin (manufactured by Maruha Corporation) was swollen in 30 ml of water and heated to dissolve.

(4) 10 g of Lf (produced by Mirai) was added to 60 ml of milk (produced by Morinaga Milk Products) heated to 50 ° C;

(5) 250 g of cream cheese (manufactured by Morinaga Milk Products Co., Ltd.) was heated and softened to a creamy state, and 50 g of granulated sugar (manufactured by Mitsui Seiko Co., Ltd.) was added and kneaded.

(6) Gelatin liquid (3), milk (4) and 1/2 lemon juice were added to cream cheese (5), mixed, and cooled.

(7) Whisk 60ml of fresh cream (Morinaga Milk Industry Co., Ltd.) and mix cream cheese It was added to (6), filled in container (2), and cooled in a refrigerator to solidify. Example 6: Preparation of Lf tablet

20 g of crystalline cellulose (manufactured by Wako Pure Chemical Industries, Ltd.) is placed in a mortar of about 1 liter (manufactured by Nakajima Seisakusho), 20 ml of water is added and mixed, and then separated by a 48-mesh sieve (manufactured by Wakashimori). Lactose (manufactured by Morinaga Milk Products Co., Ltd.) (25 g) and Pashi Lf (Morinaga Milk Products Co., Ltd.) 55 g were added and mixed. The obtained wet mass is sieved with stainless steel 20 mesh, placed on (Washamori) and extruded by hand onto two stainless steel plates for drying to form condyles, quickly and evenly distributed, and dried. The mixture was placed in a press and dried at 25 ° C for 2 days to obtain fine granules.

The dried granules are separated with a 20-mesh sieve made of polyethylene, and the granules that have passed through the sieve are spread on a wide sheet of paper, and 2 g of magnesium stearate (manufactured by Kanto Chemical Co., Ltd.) pre-sorted with 48 mesh is added. Mix to homogenize. Using a tableting machine (Kimura Seisakusho, KT-1 type 2), using an R punch with a diameter of 8 mm, the number of tablets is 10, the tablet weight is 0.62 g, and the Monsanto hardness is 3.5 to 5. Okg. Tableting was performed with the compression pressure set to obtain a 50% Lf-containing agent. The tablet was stored in a light-shielded desiccator (NRT-90B, manufactured by Nippon Rikagaku Kikai). Example 7: Preparation of encapsulated Lf hydrolyzate

Lactose (manufactured by Wako Pure Chemical Industries) 60 g, corn starch (manufactured by Nisshin Seifun Co., Ltd.) 40 g, crystalline cellulose (manufactured by Wako Pure Chemical Industries) 40 _g, and Lg hydrolyzate 60 g prepared by the same method as in Reference Example 1 Was separated by a 50-mesh sieve (manufactured by Yamato Scientific Co., Ltd.), placed in a 0.5 mm-thick polyethylene bag, mixed by inversion, and the above powder was mixed using a fully automatic capsule filling machine (manufactured by Cesere Pedini; press type). Capsules (made by Elanco Japan, No. 1 gelatin capsule, Op. Yellow No. 6 Body, empty weight: 75 mg) are filled with a content of 275 mg, and a capsule containing 82 mg of Lf hydrolyzate is obtained. Obtained. This capsule was stored at room temperature. Example 8: Preparation of solid feed containing Lf hydrolyzate

5 g of Lf hydrolyzate powder of papaya prepared by the same method as in Reference Example 1 and feed for solid rodents (Funabashi Farm Co., Ltd. F2) 1.5 kg of 0.2 mm thick transparent polyethylene Put in a bag, close the opening with a polyethylene string, shake by hand up, down, left and right for about 10 minutes, let the powder enter the fine holes of the solid sample, mix at the same time, and then 200g each in a plastic bag The solid feed containing 0.33% Lf hydrolyzate was obtained. This feed was stored in a cool and dark place. Example 9: Tablet containing Lf hydrolyzate-derived peptide

Peptides obtained from Lf hydrolysates

(Manufactured by the same method as in Reference Example 2) 50 (mg) crystalline cellulose 170

Corn starch 66

Talc 11

Magnesium stearate 3

The raw materials at the above ratios per tablet were uniformly mixed by a conventional method, granulated, dried, and tableted to obtain a tablet containing 17% of the peptide. In addition, the raw materials other than the peptide obtained from the hydrolyzate of Lf were all commercially available products. Example 10: Preparation of Bavaroi containing Lf hydrolyzate-derived peptide

Approximately 9 g of plate gelatin (manufactured by Maruha) was swollen with water, and 30 g of chocolate (manufactured by Morinaga Seika Co., Ltd. for confectionery) was shredded and melted in a water bath. Separately, put 3 egg yolks in a bowl, add 70 g of sugar (manufactured by Mitsui Seiko Co., Ltd.), stir with a whisk until the whiteness is reduced, add the molten chocolate to the mixture, and further add Stir 220 ml of milk (manufactured by Morinaga Milk Products Co., Ltd.) mixed and heated to 50 ° C is gradually added, transferred to a pan and set on low heat, heated with constant stirring, and then turned off to remove the swollen gelatin. Add, dissolve with stirring, filter with a stainless steel strainer, cool in ice water, and cool to about 50 ° C. Lf-derived peptides suspended in 30 ml of milk (Reference Example 2 and 1 g) was added, and the mixture was stirred and mixed. —One, 100ml of whipped cream (made by Morinaga Milk Co., Ltd., 45% fat whipped cream) in a bowl, whisk with a whisk, pour the mixture into the above mixture, mix quickly and pour into stainless steel mold, in refrigerator And solidified to obtain chocolate bavarois. Example 11: Powder containing Lf-derived organic synthetic peptide

Organic synthetic peptides derived from Lf

(Manufactured in the same manner as in Reference Example 3) 50 (mg) crystalline cellulose 375

Corn starch 575

The above materials were uniformly mixed per bag, and 15 bags of powder were prepared by a conventional method. In addition, the raw materials other than the organic synthetic peptide were all commercially available products. Example 12: Capsule containing organic synthetic peptide

Organic synthetic peptides derived from Lf

(Manufactured by the same method as in Reference Example 3) 10 (mg) Lactose 120

Microcrystalline cellulose 42

Carboxymethyl cellulose 10

Talc 15

Magnesium stearate 3 Each raw material in the above ratio per tablet was uniformly mixed by a conventional method, and a capsule was prepared using a capsule filling machine. In addition, the raw materials other than the organic synthetic peptide were all commercial products. Example 13: Preparation of solid feed containing Lf hydrolyzate

Using a 5 g powder of apolactoph of Azurium:!: Phosphorus hydrolyzate prepared in the same manner as in Reference Example 6, containing 0.33% of apolactoferin hydrolyzate in the same manner as in Example 8. Solid feed was obtained. This feed was stored in a cool and dark place. Example 14: Tablet Containing Peptide Derived from Apolactoferin Hydrolyzate

Peptides obtained from Apolactoferin hydrolyzate

(Manufactured by the same method as in Reference Example 6) 50 (mg) Crystal cell opening 1 7 0 Corn starch 66 Talc 1 1

Magnesium stearate 3

The raw materials at the above ratios per tablet were uniformly mixed by a conventional method, granulated, dried and tableted to obtain a tablet containing 17% peptide. In addition, all the raw materials other than the peptide obtained from the apo Lf hydrolyzate used the commercial item. Industrial applicability

The cancer metastasis inhibitor of the present invention can be easily orally ingested as foods, pharmaceuticals and the like to suppress cancer metastasis. In addition, the cancer metastasis inhibitor of the present invention contains Lf, a protein derived from milk as a food, a hydrolyzate of Lf or a peptide derived from Lf as an active ingredient. It is safe with no side effects. iH column table

SEQ ID NO: 1

Array Length: 1 1

Sequence type: amino acid

Topology: linear

Sequence type: Peptide

Sequence features: the present peptide and peptides containing the present peptide as fragments.

In the following sequence, R01 represents any amino acid residue except Cys.

Array:

Ys 01 R01 R01 R01 G in R01 R01 Met Lys Lys

1 5 10

SEQ ID NO: 2

Array Length: 1 1

Sequence type: amino acid

Topology: linear

Sequence type: Peptide

In the following sequence, R01 represents any amino acid residue except Cys.

Array:

Ys R01 R01 R01 R01 G in R01 R01 Met Arg Lys

1 5 10

SEQ ID NO: 3

Array Length: 6

Sequence type: amino acid Topology: linear

Sequence type: Peptide

In the following sequence, R01 represents any amino acid residue except Gys.

Array:

Ar g R01 R01 R01 R01 Ar g

1 5

SEQ ID NO: 4

Array Length: 6

Sequence type: amino acid

Topology: linear

Sequence type: Peptide

In the following sequence, R01 represents any amino acid residue except Cys.

Array:

Lys R01 R01 R01 R01 Ar g

1 5

SEQ ID NO: 5

Array Length: 6

Sequence type: amino acid

Topology: linear

Sequence type: Peptide

In the following sequence, R01 represents any amino acid residue except Gys Show.

Array:

Lys R01 R01 R01 R01 ys

1 5

SEQ ID NO: 6

Array Length: 6

Sequence type: amino acid

Topology: linear

Sequence type: Peptide

In the following sequence, R01 represents any amino acid residue except Cys.

Array:

Arg R01 R01 R01 R01 Lys

1 5

SEQ ID NO: 7

Array Length: 5

Sequence type: amino acid

Topology: linear

Sequence type: Peptide

In the following sequence, R01 represents any amino acid residue except Cys.

Array:

Arg R01 R01 R01 Arg

1 5 SEQ ID NO: 8

Array Length: 5

Sequence type: amino acid

Topology: linear

Sequence type: Peptide

Sequence characteristics: the present peptide, and a peptide comprising the present peptide as a fragment.

In the following sequence, R01 represents any amino acid residue except Cys.

Array:

Ys R01 R01 R01 Arg

1 5

SEQ ID NO: 9

Array Length: 5

Sequence type: amino acid

Topology: linear

Sequence type: Peptide

In the following sequence, R01 represents any amino acid residue except Cys.

Array:

Arg R01 R01 R01 then ys

1 5

SEQ ID NO: 10

Array Length: 6

Sequence type: amino acid

Topology: linear Sequence type: Peptide

Sequence characteristics: the present peptide and a peptide containing the present peptide as a fragment

Phe Gin T r p Gin A r g As n

1 5

SEQ ID NO: 1 1

Array Length: 5

Sequence type: amino acid

Topology: linear

Sequence type: Peptide

Sequence characteristics: the present peptide and a peptide containing the present peptide as a fragment ₍ sequence:

Phe Gin Trp Gin Ar g

1 5

SEQ ID NO: 12

Array Length: 4

Sequence type: amino acid

Topology: linear

Sequence type: Peptide

Sequence features: the present peptide and peptides containing the present peptide as fragments. Array:

Gin Trp Gin Ar g

1

SEQ ID NO: 13

Array Length: 3

Sequence type: amino acid Topology: linear

Sequence type: Peptide

Sequence characteristics: the present peptide and a peptide containing the present peptide as a fragment, sequence:

Trp Gin Arg

1

SEQ ID NO: 14

Array Length: 5

Sequence type: amino acid

Topology: linear

Sequence type: Peptide

Sequence characteristics: the present peptide, and a peptide containing the present peptide as a fragment.

Ar Ar Trp Gin Trp

1 5

SEQ ID NO: 15

Array Length: 4

Sequence type: amino acid

Topology: linear

Sequence type: Peptide

Arg Ar Trp Gin

1

SEQ ID NO: 16

Array Length: 4 Sequence type: amino acid

Topology: linear

Sequence type: Peptide

Trp Gin T r p A r g

1

SEQ ID NO: 17

Array Length: 3

Sequence type: amino acid

Topology: linear

Sequence type: Peptide

Gin Trp Arg

1

SEQ ID NO: 18

Array Length: 6

Sequence type: amino acid

Topology: linear

Sequence type: Peptide

Leu ftr g Trp Gin As n Asp

1 5

SEQ ID NO: 19 Array Length: 5

Sequence type: amino acid

Topology: linear

Sequence type: Peptide

Leu A r g T r p Gin As n

1 5

SEQ ID NO: 20

Array Length: 4

Sequence type: amino acid

Topology: linear

Sequence type: Peptide

Leu A r g Trp Gin

1

SEQ ID NO: 21

Array Length: 3

Sequence type: amino acid

Topology: linear

Sequence type: Peptide

Arg Trp Gin

1 SEQ ID NO: 22

Array length: 20

Sequence type: amino acid

Topology: linear

Sequence type: Peptide

In the following sequence, Cys 2 and Cys 19 are disulfide bonded.

Array:

Ly s Cys Ar g A r g T rp Gin T rp Ar g Met L s Ly s Leu G ι y Ala Pro 1 5 10 15

Ser l ie Thr Cys Va I

20

SEQ ID NO: 23

Array length: 20

Sequence type: amino acid

Topology: linear

Sequence type: Peptide

In the following sequence, Cys * indicates a cysteine in which the thiol group has been chemically modified to prevent the formation of disulfide bonds. Array:

Ly s Cys * Ar g Ar g T rp Gin Trp Ar g Met Ly s Lys Leu G I y A I a Pro 1 5 10 15 Ser Me Thr Cys * Va I

20 SEQ ID NO: 24

Array length: 20

Sequence type: amino acid

Topology: linear

Sequence type: Peptide

In the following sequence, Cys No. 2 and Cys No. 19 are disulphide yarns o σ しし ο ο

Array:

Ys Cys Phe Gin T rp Gin A rg Asn Met Ar g ys Va I A rg G I y Pro

1 5 10 15

Pro Va I Ser Cys I I e

20

SEQ ID NO: 25

Array length: 20

Sequence type: amino acid

Topology: linear

Sequence type: Peptide

In the following sequence, C * indicates a cysteine in which the thiol group has been chemically modified to prevent the formation of disulfide bonds. Array:

Ly s Cys * Phe Gin T rp Gin Ar g Asn Met Ar g Ly s Va I Ar g G I y Pro 1 5 10 15 Pro Va I Ser Cys * I I e

20 SEQ ID NO: 26

Array Length: 25

Sequence type: amino acid

Topology: linear

Sequence type: Peptide

In the following sequence, Gys 3 and Cys 20 are disulfide bonded.

Array:

Phe Ly s Cys Ar g Ar g I'rp Gin T rp A r g Met Ly s Ly s Leu G I y Ala

1 5 10 15

Pro Se r lie Th r Cys Va I Ar g Ar g A I a Phe

20 25

SEQ ID NO: 27

Array Length: 1 1

Sequence type: amino acid

Topology: linear

Sequence type: Peptide

Sequence characteristics: the present peptide, and a peptide containing the present peptide as a fragment

Array:

Ly s Th r Ar g Ar g i'rp Gin Trp Ar g Met Ly s Ly s

1 5 10

SEQ ID NO: 28

Array Length: 38

Sequence type: amino acid Topology: linear

Sequence type: Peptide

In the following sequence, Cys # 16 and Cys # 33 are disulfide bonded.

Array:

Ys As n Va I Arg T r p Cys Th r I I e Ser Gin Pro G I uT rp Phe ys 1 5 10 15

Cys Ar g A r g T r p Gi n T r p Ar g Met Ly s ys Leu G I y A I a Pro Ser

20 25 30 l ie Thr Cys Va I Ar g Ar g A I a Phe

35

SEQ ID NO: 29

Array length: 32

Sequence type: amino acid

Topology: linear

Sequence type: Peptide

In the following sequence, Cys No. 10 and Cys No. 27 are disulfide bonded.

Array:

Thr lie Ser Gin Pro Glu T r p Phe Lys Cys Ar g Ar g T r p Gin Tr p 1 5 10 15

Ar g Met Ly s Ly s Leu G I y Ala Pro Ser I I e Thr Cys Va I Ar g Ar g

20 25 30

Ala Phe SEQ ID NO: 30

Array length: 47

Sequence type: amino acid

Topology: linear

Sequence type. 'Peptide

In the following sequence, the peptide having the sequence length of 36 and having Cys at positions 9, 26, and 35 has a disulfide bond between Gys 9 and Cys 26, and the length of the sequence 36 No. 35 of the peptide has a Cys force of <<<<, the sequence length is "M", and the peptide having Cys at No. 10 has a disulfide bond with Cys at No. 10.

Array:

Va I Ser Gin Pro Glu A I a Thr Lys Cys Phe Gin Trp Gin Ar Asn 1 5 10 15 Met Ar g Lys Va I Ar g G I y Pro Pro 'Va I Ser Cys I I e Lys Ar g Asp

20 25 30

Ser Pro l ie Gin Cys l ie

35

G I y Ar g Ar g Ar g Ar g Ser Va I Gin Trp Cys A I a

1 5 10

SEQ ID NO: 31

Array Length: 5

Sequence type: amino acid

Topology: linear

Sequence type: Peptide

Sequence characteristics: the present peptide, and a peptide comprising the present peptide as a fragment. In the following sequence, R01 represents any amino acid residue except Cys.

Array:

Lys R01 R01 R01 Lys

1 5

Claims

The scope of the claims

1. Non-ferrous saturated lactoferrin, hydrolysates of lactoferrins, pharmaceutically acceptable derivatives of the hydrolysates, pharmaceutically acceptable salts of the hydrolysates, lactofu :! : One or two substances selected from the group consisting of peptides derived from a hydrolyzate of phosphorus, a pharmaceutically acceptable derivative of the peptide, and a pharmaceutically acceptable salt of the peptide. An oral cancer metastasis inhibitor contained as an active ingredient.

2. Non-ferrous saturated lactoferrin, a hydrolyzate of lactoferrins, a pharmaceutically acceptable derivative of the hydrolyzate, or a pharmaceutically acceptable salt of the hydrolyzate is 3 to 3200 mgZ. 3. The oral cancer metastasis inhibitor according to claim 1, which is orally administered at a weight of kg.

3. A peptide derived from a hydrolyzate of lactoferrin, a pharmaceutically acceptable derivative of the peptide, or a pharmaceutically acceptable salt of the peptide is 0.2

The oral cancer metastasis inhibitor according to claim 1, which is orally administered at a rate of 〜320 mgZ.

4. The oral cancer metastasis inhibitor according to claim 1 or 3, wherein the peptides have the amino acid sequence of any one of SEQ ID NOs: 1 to 31.

5. A food composition comprising the oral cancer transfer inhibitor according to any one of claims 1 to 4 as an essential component.

6. A feed composition comprising the oral cancer transfer inhibitor according to claim 1 as an essential component.