JPH05271094A - Cancer metastasis-inhibiting agent using carboxymethylated chitin derivative - Google Patents

Abstract

PURPOSE:To provide the subject medicinal agent containing a carboxymethylated chitin derivative having a peptide sequence comprising an arginine residue- glycine residue-aspartic acid residue on its side chain as an active ingredient. CONSTITUTION:Amino acids whose a-amino groups are protected with t- butoxycarbonyl (Boc) groups are successively bonded to a solid phase carrier resin according to the amino acid sequence of a peptide by a peptide solid phase synthesis using a peptide-synthesizing device of Merrifield style, and the product is subjected to the removal of the protecting groups and to the release of the peptide chain from the resin to obtain the adhesive peptide containing the peptide sequence comprising the arginine residue-glycine residue- aspartic acid residue. Subsequently, the peptide is reacted with a carboxymethylated chitin in the presence of a water-soluble carbodiimide as a condensation agent, etc., in a buffer solution to obtain the cancer metastasis- inhibiting agent comprising the carboxymethylated chitin derivative containing the cell-adhesive peptide units.

Description

Detailed Description of the Invention

[0001]

The present invention relates to Arg-Gly-Asp.
The present invention relates to a cancer metastasis inhibitor comprising a carboxymethylated chitin derivative (hereinafter abbreviated as “CM-chitin derivative”) having the tripeptide as an essential unit and a pharmacologically acceptable salt thereof as an active ingredient.

[0002]

2. Description of the Related Art Fibronectin is a protein involved in cell-extracellular matrix adhesion and is also considered to be involved in platelet aggregation and cancer metastasis. These interactions are mediated by a series of cell surface receptors. Although fibronectin is a macromolecule with a molecular weight of about 250,000, these receptors
It was revealed that the g-Gly-Asp sequence was specifically recognized, and it was reported that it is important for the interaction with the receptor (Nature, Vol. 309, 30).
Page, 1984). This Arg-Gly-Asp sequence is also present in other adhesive proteins such as vitronectin, and through the core sequence, it binds to the receptor of adherent cells and transmits the information to adherent cells. Also, heparin, collagen,
It also has the ability to bind to biopolymers such as fibrin, and is also considered to be involved in adhesion between cells and stromal connective tissue, cell differentiation, and proliferation. As described above, since the cell adhesion-activating protein has various biological activities, its application to medicines and medical materials has been studied. For example, a method of inhibiting platelet aggregation using various linear and cyclic oligopeptides having Arg-Gly-Asp sequence (Polymer Prepr
ints, Japan), Volume 38, page 3149, 1989, JP-A 2-1747
97), a method of using a peptide having an Arg-Gly-Asp sequence as a cell migration inhibitor (Japanese Patent Laid-Open No. 2-4716), Arg-Gly-
Method of using Asp-immobilized PMMA membrane as cell adhesion membrane
(Polymer Preprints, Japan), 37th
Vol., P. 705, 1988). Further, a method of covalently bonding a peptide having Arg-Gly-Asp as an essential constituent unit to a polymer and using it as a substrate for animal cell culture or a substrate for biocomposite artificial organs (JP-A-1-309682, JP-A-1-305960)
No.), and a method of using a polypeptide having an Arg-Gly-Asp-Ser sequence as a platelet protecting agent for extracorporeal blood (JP-A-64-6217). Furthermore, cell adhesion activating proteins are also attracting attention as substances related to cancer metastasis. During a series of stages of cancer metastasis, cancer cells come into contact with various host cells and biopolymers. At this time, when a cell adhesion molecule such as fibronectin is present, the cells form a multicellular mass, which facilitates the growth and survival of cancer cells. However, when the above-mentioned cell adhesive activity protein is present, the tripeptide Arg-Gly-Asp, which is an adhesion core of fibronectin, contained in the protein.
Has been reported to exhibit cancer metastasis-inhibiting activity by binding to a receptor on cancer cells (Science, 238, 467, 1986). Furthermore, a method for more efficiently suppressing cancer metastasis using an oligopeptide having this sequence or a polypeptide having a repeating structure thereof is also disclosed (Int. J. Biol. Macr.
omol., vol. 11, p. 23, 1989; ibid., vol. 11, p. 226,
1989; Jpn. J. Cancer Res., 60, 722, 1989.
Year). On the other hand, chitin is a polysaccharide in which N-acetyl-D-glucosamine is β- (1 → 4) linked, and is a main component of the exoskeleton of crustaceans and insects. It is widely distributed in lower animals and invertebrates, plays a role in supporting and protecting the living body, and corresponds to cellulose in the plant kingdom. Chitin is also called the last biomass, and studies on its derivatives have been actively conducted in recent years, and many studies on chitin derivatives which are soluble in a solvent have been particularly reported. Carboxymethyl chitin (CM-chitin) obtained by carboxymethylating the hydroxyl group at the C-6 position of N-acetyl-D-glucosamine unit is water-soluble and is an important compound as a starting material for various chitin derivatives. And C
The CM-chitin derivative in which the hydroxyl group at the C-6 position or the C-3 position of M-chitin is sulfated is sulfated carboxymethyl chitin (SCM-chitin). Regarding these chitins and their derivatives, "application of chitin-chitosan" edited by Chitin-Chitosan Study Group (Gihodo, 1990), "last biomass Chitin-Chitosan" edited by Chitin-Chitosan Research Group (Gihodo, 1988), It is described in detail in the Chitin-Chitosan Experiment Manual (Gihodo, 1991). In addition, heparan sulfate, which is one of the important components of extracellular matrix and basement membrane, is decomposed by heparanase produced from cancer metastatic melanoma cells during the wetting process of cancer, and this enzymatic decomposition is inhibited by heparin. (Science, No. 220)
Volume, 611, 1984; J. Biol. Chem., 59th.
Vol., 2283, 1984). Heparin and heparin derivatives, which have no anticoagulant effect, have been found to suppress cancer metastasis (GANN Monograph of Can.
cer Res., Volume 20, 147, 1977; Bioche.
m., 25, 5322, 1986), SCM-chitin having a heparin-like structure is also known to suppress cancer metastasis (Jpn. J. of Cancer Res.,
Volume 80, 866, 1989; Cancer Res., 5th.
0, 3631, 1990; Cancer Res., 51st.
Vol. 22, p. 1991). As mentioned above, Arg-Gly-As
Although it has been reported that p-tripeptide, SCM chitin derivative and the like have cancer metastasis-suppressing action, it is a derivative of CM-chitin
The application of a compound introduced with an oligopeptide containing Arg-Gly-Asp as an essential unit or a polypeptide having a repeating structure thereof to a cancer metastasis inhibitor is not known at all.

[0003]

As described above, cell adhesion-activating proteins such as fibronectin or peptide fragments thereof have various biological activities, and it is desired to develop a technique for applying the related substances as a medicine. Was there. In particular,
The cancer metastasis suppressive action of the adhesive core sequence is considered to have high application value as a medicine. However, the tripeptide of the adhesion core sequence itself undergoes various metabolic actions in vivo and is likely to be excreted out of the body at an early stage, and its concentration in blood cannot be stably maintained. It was difficult to get the effect. Therefore, the present inventors have focused on the fact that natural polysaccharides have various properties and functions, and the interaction with living organisms is very different from that of low molecular weight compounds and other polymers. As a result of earnest research for a novel compound that retains the biological activity of sucrose, is easy to synthesize and has no serious side effects on the living body, a new compound having the tripeptide Arg-Gly-Asp as an essential unit This study has been completed by discovering various CM-chitin derivatives and their pharmacologically acceptable salts. Therefore, an object of the present invention is to enhance the ability to bind to the receptor, have high stability in blood, and produce Arg-Gly-Asp that can be produced by a simpler method.
Novel CM-having the tripeptide of
It is intended to provide a pharmaceutical composition containing a chitin derivative and a pharmacologically acceptable salt thereof.

[0004]

[Means for Solving the Problems] The pharmaceutical composition of the present invention comprises:
A cancer metastasis inhibitor comprising, as an active ingredient, a CM-chitin derivative having a peptide sequence consisting of an arginine residue-glycine residue-aspartic acid residue in its side chain or a pharmacologically acceptable salt thereof. The tripeptide sequence may be present as part of a peptide sequence containing other amino acids, and the CM-chitin derivative may be provided with an organic group such as an alkylene group or an arylene group that does not affect the physiological activity of the compound. It may be coupled via. Further, in consideration of the production of the compound, it may be bonded via an amide bond, an ester bond, an ether bond, a urethane bond or the like. The non-bonded end of the group containing the above tripeptide sequence may have an alkyl group, an aryl group or the like. In a particularly preferred embodiment, the cancer metastasis inhibitor of the present invention has the following general formula [1] bonded to a side chain of a carboxymethylated chitin derivative through any of an amide bond, an ester bond, an ether bond and a urethane bond. ] A peptide sequence-carrying carboxymethylated chitin derivative having an adhesive peptide represented by the following as an essential unit or a pharmacologically acceptable salt thereof is contained as an active ingredient. [R ¹ ]-[CO]-([X] -Arg-Gly-Asp- [Y]) n- [Z]-[R ² ] [1] In the formula, Arg is arginine, Gly is glycine, and Asp is Indicates an aspartic acid residue. [] Represents that the group in [] may or may not be present, and when it is present, X and Y are respectively serine (Ser), glycine (Gly), valine (Val), Asparagine (As
n), aspartic acid (Asp), proline (Pro)
Represents a amino acid residue selected from or a peptide residue composed of these amino acid residues, and Z represents -O- or -NH-. One of R ¹ and R ² is hydrogen,
Alternatively, a linear or branched alkyl group having 1 to 9 carbon atoms or an aryl group having 6 to 9 carbon atoms, which may have a substituent and may contain an unsaturated bond,
The other represents a linear or branched alkylene group having 1 to 9 carbon atoms, which may have a substituent or may contain an unsaturated bond, or an arylene group having 6 to 9 carbon atoms. .. n shows the integer of 1-5. Examples of the substituent of the alkyl group and the alkylene group include a carbonyl group, a carboxyl group, an amino group, a hydroxyl group, a sulfo group, a halogen atom, an aryl group, a nitro group, and a cyano group.
You may have two or more in the same chain. The unsaturated bond may be a double bond or a triple bond. Preferred CM-used in the peptide sequence-carrying CM-chitin derivative of the present invention
Chitin derivatives include sulfated carboxymethyl chitin (SCM-chitin), carboxylated CM-chitin and CM-chitin. The peptide sequence-carrying CM-chitin derivative used in the present invention preferably has a molecular weight of 2
It is preferably water-soluble at room temperature in the range of 0,000 or less, particularly in the range of 3,000 to 100,000. Examples of the carboxylating agent for carboxylated chitin include succinic anhydride, maleic anhydride, phthalic anhydride, itaconic anhydride, citraconic anhydride, pyromellitic dianhydride, trimellitic dianhydride, and the like. The amino acid used in the cell-adhesive peptide according to the present invention may be L-form or D-form, but is preferably L-form. Examples of the salt of the peptide sequence-supporting CM-chitin derivative of the present invention include salts with inorganic acids such as hydrochlorides, sulfates, nitrates, phosphates and borates, and acetates,
Examples thereof include salts with organic acids such as trifluoroacetates, trifluoromethanesulfonates, lactates, and tartrates, and conversion into such salts can be carried out by a conventional means. The peptide synthesis method is not particularly limited, and examples thereof include a liquid phase method, a solid phase method, and a synthetic method using an automatic synthesizer. For details of these synthetic methods, see Biochemistry Experiment Course "Protein Chemistry IV" p207-495 (edited by the Japanese Biochemical Society, Tokyo Kagaku Dojin), "Sequence Biochemistry Experimental Course Protein Chemistry (below)" (Japan Biochemical Society) Ed., Tokyo Kagaku Dojin), Izumiya et al., "Basics and Experiments of Peptide Synthesis" (Maruzen). It is also possible to use a commercially available synthetic peptide. Examples of the method used for binding the CM-chitin derivative to the adhesive peptide include an amide bond synthesizing method using cyanogen bromide, acid azide, water-soluble carbodiimide and the like. The peptide sequence-carrying CM-chitin derivative of the present invention has a core sequence Arg-Gly-Asp of a cell adhesive protein, and adheres to cells through the core sequence by a mechanism similar to that of the cell adhesive protein. Therefore, it shows various biological activities as an agonist or antagonist of cell adhesive protein, and has a wide range of biological activities such as immunomodulatory action, wound healing action, platelet aggregation inhibitory action by cancer cells occurring in capillaries, and neurological disease healing action. Is recognized. Therefore, the peptide sequence-carrying CM-chitin derivative of the present invention is used not only as a cancer metastasis inhibitor, but also as a wound healing agent, an immunomodulator, a platelet aggregation agent, together with a conventional carrier or a pharmaceutical aid, if necessary. It can be administered to a patient as an anti-adhesion agent. The dose is in the range of 0.2 μg / kg to 400 mg / kg,
It is determined based on symptoms, age, weight, etc. The chitin derivative of the present invention is preferably administered by an administration method generally used for peptide drugs, that is, a parenteral administration method such as intravenous administration, intramuscular administration, subcutaneous administration and the like. In the case of producing such an injectable preparation, the chitin derivative of the present invention may be dissolved in PBS or physiological saline to give an injectable preparation, as shown in the Examples below, or an aqueous solution of about 0.1N acetic acid. And the like, and then may be used as a freeze-dried preparation. A conventional stabilizer such as glycine or albumin may be added to such a preparation. Furthermore, the chitin derivative of the present invention or a salt thereof can be orally administered, for example, in the form of microcapsules encapsulated in liposomes, microspheres, or hydrogels.
Further, in the form of suppository, sublingual tablet, nasal spray, etc., it can be absorbed from mucous membranes other than digestive tract.

[Examples] The present invention will be further described with reference to the following examples, but the present invention is not limited thereto.
Amino acids, various protecting groups, deprotection reagents and the like are represented by commonly used abbreviations. The main abbreviations used for the notation of protecting groups and deprotection reagents are as follows. Bzl: benzyl group t-Boc: t-butoxycarbonyl group Z: benzyloxycarbonyl group Pac: phenacyl group Ac: acetyl group TFA: trifluoroacetic acid HOBt: 1-hydroxybenzotriazole DCC: dicyclohexylcarbodiimide DC urea: dicyclohexylurea AcOEt: Ethyl acetate

Production Example 1 Synthesis of Adhesive Peptide by Solid Phase Method Synthesis was carried out using a peptide synthesizer by the Merrifield method. The Boc group was used for the protection of the α-amino group, and the peptide was cleaved from the resin and then purified by preparative HPLC (high performance liquid chromatography) to obtain an adhesive synthetic peptide showing a single peak.

Table 1 Adhesive synthetic peptides ───────────────────────────────── Name Structural formula Abbreviation Yield ── ─────────────────────────────── Peptide-1 H-Arg-Gly-Asp-OH RGD 37% Peptide-2 H -(Arg-Gly-Asp) ₂ -OH (RGD) ₂ 28% peptide-3H- (Arg-Gly-Asp) ₃ -OH (RGD) ₃ 19% peptide-4H- (Arg-Gly-Asp) ₅ -OH (RGD) ₅ 11% ─────────────────────────────────

Production Example 2 Synthesis of Adhesive Peptide RGD by Liquid-Phase Method Boc-Asp (Purchased from Domestic Chemical Co., Ltd.) by the method described in the literature (Chem. Pharm. Bull., 24, 3025 (1976)). OBzl)
32.3 g, triethylamine 14 ml, benzyl bromide 17.1
A mixture of g and 200 ml of ethyl acetate was heated under reflux for 3 hours.
The reaction solution was allowed to cool to room temperature, washed with 1N aqueous sodium hydrogen carbonate solution and saturated saline (200 ml each), and dried over anhydrous sodium sulfate. Sodium sulfate was removed by filtration, and the filtrate was concentrated under reduced pressure to give a colorless oil. The reaction mixture was purified by silica gel chromatography (eluent hexane / ethyl acetate 95: 5), Boc-Asp (OBzl) -O.
36 g of Bzl were obtained. Dissolve in 20 ml of dichloromethane, add 20 ml of trifluoroacetic acid and stir for 30 minutes at room temperature.
(OBzl) -OBzl was obtained quantitatively. TFA-Asp (OBzl) -OBzl 8.5
g in dichloromethane, Boc-Gly anhydrous 6.7 g,
2.5 g of dimethylaminopyridine (DMAP) was added, and the mixture was stirred at room temperature for 6 hours. The reaction solution was washed with water and dried over anhydrous sodium sulfate. Sodium sulfate was removed by filtration, and the filtrate was concentrated under reduced pressure. The residue was dissolved in 20 ml of dichloromethane, 20 ml of trifluoroacetic acid was added, and the mixture was stirred at room temperature for 30 minutes. After the solvent was distilled off under reduced pressure, 100 ml of chloroform was added, and the mixture was washed several times with 100 ml of 1N sodium hydrogen carbonate aqueous solution and 100 ml of saturated saline each time, and dried over anhydrous sodium sulfate. Sodium sulfate was filtered off and the filtrate was concentrated under reduced pressure to give Gly-Asp (OBzl) -OBzl.
Got Boc-Arg (Z) ₂ (purchased from Kokusan Kagaku Co., Ltd.) 10.83g, DCC 4.54g, HOB without purification
t 2.76 g and DMF 80 ml were added and the mixture was stirred for 24 hours to remove DC urea, and then the solvent was distilled off under reduced pressure.
0 ml was added, the mixture was washed with 1N aqueous sodium hydrogen carbonate solution and 200 ml each of saturated saline and dried over anhydrous sodium sulfate. Sodium sulfate was removed by filtration, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent chloroform / ethyl acetate 6: 4) to give Boc-Arg (Z) ₂ -Gly-Asp (OBzl) -OBzl 13.2g as a white powder.
Got FAB-MASS (M + H) ⁺ 894. 1.34 g of Boc-Arg (Z) ₂ -Gly-Asp (OBzl) -OBzl was dissolved in 10 ml of methylene chloride, 10 ml of trifluoroacetic acid was added, and the mixture was stirred at room temperature.
Stir for 30 minutes. The solvent was distilled off under reduced pressure. Peptide-1 H-Arg-Gly- was added to the residue by adding ethyl acetate and adding 5% palladium-carbon (100 mg) for hydrogenolysis (room temperature, 8 hours), filtering and concentrating the reaction solution, followed by ion exchange treatment. Asp
-OH was obtained quantitatively.

Preparation of Boc-Asp (OBzl) -OPac 16.2 g (50 mmol) of Boc-Asp (OBzl), triethylamine (Et
5.05 g (50 mmol) of ₃ N) was dissolved in 50 ml of DMF and stirred, and 50 ml of a DMF solution containing 9.95 g (50 mmol) of phenacyl bromide was added dropwise under ice cooling. After returning to room temperature and stirring for 4 hours to distill off DMF under reduced pressure, the residue was washed with ethyl acetate (A
It was dissolved in cOEt) and washed with 10% aqueous citric acid solution. Subsequently, the organic layer was washed with a 10% aqueous sodium hydrogen carbonate solution, the organic layer was dried over sodium sulfate and then filtered, and the filtrate was concentrated under reduced pressure. Recrystallize the residue with AcOEt-hexane system
18.0 g (41 mmol, yield 82%) of Boc-Asp (OBzl) -0Pac was obtained as a colorless powder.

Production Example 3 Synthesis of Cell Adhesive Peptide RGDS Boc-Ser (Bzl) 2 purchased from Kokusan Kagaku Co., Ltd. by the method described in the literature (Chem. Pharm. Bull., 24, 3025 (1976)).
9.5 g, triethylamine 14 ml, benzyl bromide 17.1 g,
A mixture of 200 ml of ethyl acetate was heated to reflux for 3 hours. The reaction solution was allowed to cool to room temperature, washed with 1N aqueous sodium hydrogen carbonate solution and saturated saline (200 ml each), and dried over anhydrous sodium sulfate. Sodium sulfate was removed by filtration, and the filtrate was concentrated under reduced pressure to give a colorless oil. The reaction mixture was purified by silica gel chromatography (eluent hexane / ethyl acetate 40: 1) to give Boc-Ser (Bzl) -OBzl 3
Obtained 6 g. Next, 7.71 g of Boc-Ser (Bzl) -OBzl was dissolved in 20 ml of methylene chloride, 20 ml of trifluoroacetic acid was added, and the mixture was stirred at room temperature for 30 minutes. After the solvent was distilled off under reduced pressure, 100 ml of chloroform was added, and the mixture was washed several times with 100 ml of a 1N sodium hydrogen carbonate aqueous solution and 100 ml of a saturated saline solution and dried over anhydrous sodium sulfate. Sodium sulfate was removed by filtration, and the filtrate was concentrated under reduced pressure to give a colorless oil. This and Boc-Asp (OBzl) (domestic chemical
A mixture of 6.47 g (purchased from KK), dicyclohexylcarbodiimide (DCC) 4.54 g, hydroxybenzotriazole (HOBt) 2.76 g and DMF 80 ml was stirred at 0 ° C. for 3 hours and further at room temperature for 12 hours. After removing the DC urea, the solvent was distilled off under reduced pressure, 100 ml of chloroform was added, and 1
The solution was washed with 200 ml each of an aqueous sodium hydrogencarbonate solution and saturated saline solution, and dried over anhydrous sodium sulfate. Sodium sulphate is filtered off and the filtrate is concentrated under reduced pressure and diluted with dichloromethane 20
ml and trifluoroacetic acid 20 ml were added and the mixture was stirred at room temperature for 30 minutes. After distilling off the solvent under reduced pressure, 100 ml of chloroform was added, and 100 ml of 1N sodium hydrogen carbonate aqueous solution and saturated saline solution were added.
It was washed several times with and dried over anhydrous sodium sulfate. Sodium sulfate was removed by filtration, the filtrate was concentrated under reduced pressure, and Asp (OBz
l) -Ser (Bzl) -OBzl was obtained. In addition, Boc-Gly (domestic chemical
Purchased from Co., Ltd.) 3.50g, DCC 4.54g, HOBt 2.76
g, 80 ml of DMF was added, and the mixture was kept at 0 ° C for 3 hours and at room temperature for 1
Stir for 2 hours. After removing the DC urea, the solvent was distilled off under reduced pressure, 100 ml of chloroform was added, the mixture was washed with 200 ml of a 1N sodium hydrogen carbonate aqueous solution and 200 ml of a saturated saline solution, and dried over anhydrous sodium sulfate. Sodium sulfate is filtered off,
The filtrate was concentrated under reduced pressure, 20 ml of dichloromethane and 20 ml of trifluoroacetic acid were added, and the mixture was stirred at room temperature for 30 minutes. After the solvent was distilled off under reduced pressure, 100 ml of chloroform was added, and the mixture was washed several times with 100 ml of 1N sodium hydrogen carbonate aqueous solution and 100 ml of saturated saline each time, and dried over anhydrous sodium sulfate. Sodium sulfate was removed by filtration, and the filtrate was concentrated under reduced pressure to Gly-Asp (OBzl) -Ser (Bzl) -O.
I got Bzl. Boc-Arg (Z) without purification
₂ (Purchased from Domestic Chemicals Co., Ltd.) 10.83g, DCC 4.54g, H
Add 2.76 g of OBt and 80 ml of DMF and stir all day and night.
After removing C urea, the solvent was distilled off under reduced pressure, 100 ml of chloroform was added, the mixture was washed with 200 ml of 1N sodium hydrogen carbonate aqueous solution and 200 ml of saturated saline solution, and dried over anhydrous sodium sulfate. Sodium sulfate was removed by filtration, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent chloroform / methanol 99: 1) to give Boc-Arg (Z) ₂ -Gly-Asp (OBzl) -Ser as a white powder.
(Bzl) -OBzl 14.4 g was obtained. FAB-MASS (M + H) ⁺ 1071. 1.03 g of Boc-Arg (Z) ₂ -Gly-Asp (OBzl) -Ser (Bzl) -OBzl was dissolved in 10 ml of methylene chloride, 10 ml of trifluoroacetic acid was added and the mixture was stirred at room temperature for 30 minutes, and then the solvent was distilled off under reduced pressure. Left. Peptide-5H-Ar was added to the residue by adding ethyl acetate and adding 100% of 5% palladium on carbon for hydrogenolysis (room temperature, 8 hours), filtering and concentrating the reaction solution, followed by ion exchange treatment.
g-Gly-Asp-Ser-OH was obtained quantitatively.

Production Example 4 Synthesis of Peptide-6 H-Gly-Arg-Gly-Asp-Ser-Pro-OH by Solid Phase Method Synthesis was carried out using a peptide synthesizer by the Merrifield method. The Boc group was used for the protection of the α-amino group, and the peptide was cleaved from the resin and then purified by preparative HPLC (high performance liquid chromatography) to obtain an adhesive synthetic peptide showing a single peak. Yield 25%.

Production Example 5 Synthesis of Peptide-2 by Liquid Phase Method 15.4 g (35 mmol) of Boc-Asp (OBzl) -OPac in dichloromethane (C
H ₂ Cl ₂ ) dissolved in 50 ml, trifluoroacetic acid (TFA) 50
ml was added under ice cooling, the temperature was returned to room temperature, and the mixture was stirred for 30 minutes. The solvent was evaporated under reduced pressure, and the residue was crystallized from ether (Et ₂ O) and Asp (OBz
17.1 g of l) -OPac TFA salt was obtained. Next, Boc-Gly 7.88g
(45 mmol), N-methylmorpholine (NMM) 4.55 g (45 mmol)
It was dissolved in 30 ml of HF and stirred. At -10 ° C, 10 ml of a THF solution of chloroformate formate (IBCF) was added and stirred.
After 5 minutes, 13.6 g (30 mmol) of Asp (OBzl) -OPacTFA salt was added, and then 10 ml of a THF solution of 3.03 g (30 mmol) of NMM was added. After reacting for 4 hours, the solvent was distilled off under reduced pressure, the residue was dissolved in AcOEt and washed with a 10% aqueous citric acid solution. Subsequently, the organic layer was washed with a 5% aqueous sodium hydrogen carbonate solution and distilled water, the organic layer was dried over sodium sulfate and then filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography (eluent AcOEt / hexane = 1/1) and Boc-Gly-Asp (OBzl) -OPac 13.6 g (27 mmol, yield 91)
%) Boc-Gly-Asp (OBzl) -OPac 13.6 g (27 mmol) in C
Dissolved in 50 ml of H ₂ Cl ₂ and trifluoroacetic acid (TF
A) 50 ml was added, and the mixture was returned to room temperature and stirred for 1 hour. The solvent was distilled off under reduced pressure and the residue was crystallized with diethyl ether (Et ₂ O) and Gl
The TFA salt of y-Asp (OBzl) -OPac was obtained quantitatively. Then Bo
c-Arg (Mts) 20.5 g (45 mmol), N-methylmorpholine (NMM)
4.55 g (45 mmol) was dissolved in 30 ml of THF and stirred. -1
10 ml of THF solution of chloroformate formate (IBCF) at 0 ℃
Was added and stirred. After 5 minutes, Gly-Asp (OBzl) -OPacTFA salt
After adding 14.8 g (29 mmol), NMM 3.03 g (30 mmol) T
10 ml of HF solution was added. After reacting overnight, the solvent was distilled off under reduced pressure, the residue was dissolved in AcOEt, and washed with 10% aqueous citric acid solution. Subsequently, the organic layer was washed with 5% aqueous sodium hydrogen carbonate solution and brine, the organic layer was dried over sodium sulfate and then filtered, and the filtrate was concentrated under reduced pressure. Silica gel chromatography (eluent AcOEt)
Purified with Boc-Arg (Mts) -Gly-Asp (OBzl) -OPac 17.1g (21
mmol, yield 71%) was obtained. Boc-Arg (Mts) -Gly-Asp (OBzl)-
5.0 g (6.0 mmol) of OPac was dissolved in 132 ml of 90% acetic acid aqueous solution and stirred, and 11.7 g of zinc powder was added under ice cooling, and the mixture was stirred at room temperature for 1.5
Reacted for hours. After that, the solvent was distilled off under reduced pressure to obtain a residue.
A 10% aqueous citric acid solution was added, and the mixture was extracted with chloroform (CHCl ₃ ). The organic layer was dried over sodium sulfate and then filtered,
The filtrate was concentrated under reduced pressure. The residue was recrystallized with AcOEt-hexane system and Boc-Arg (Mts) -Gly-Asp (OBzl) 2.56g (3.6mmo
l, yield 60%) was obtained as a colorless powder. Next, Boc-A
2.56 g (3.6 mmol) of rg (Mts) -Gly-Asp (OBzl) in 50 ml of THF
Dissolved in Arg (Mts) -Gly-Asp (OBzl) -OPac hydrochloride 2.77g
(3.6 mmol), diphenylphosphoryl azide (DPPA)
0.99 g (3.6 mmol) and triethylamine 0.36 g (3.6 mmol) were added, and the mixture was reacted overnight. After that, the solvent was distilled off under reduced pressure,
The residue was dissolved in AcOEt and washed with 10% aqueous citric acid solution. Subsequently, the organic layer was washed with 5% aqueous sodium hydrogen carbonate solution and brine, the organic layer was dried over sodium sulfate and then filtered, and the filtrate was concentrated under reduced pressure. Recrystallize the residue (Ac
OEt / hexane) and then Boc-Arg (Mts) -Gly-Asp (OBzl) -Arg (M
4.5 g (3.13 mmol, yield 87%) of ts) -Gly-Asp (OBzl) -OPac was obtained. Boc-Arg (Mts) -Gly-Asp (OBzl) -Arg (Mts) -Gly-Asp
4.3 g (3.0 mmol) of (OBzl) -OPac was dissolved in 100 ml of 90% acetic acid aqueous solution and stirred, and 9.75 g of zinc powder was added under ice cooling, and the mixture was reacted at room temperature for 1.5 hours. Then, the solvent is distilled off under reduced pressure,
To the residue was added 10% aqueous citric acid solution, and chloroform (CH
And extracted with Cl _3). The organic layer was dried over sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. AcOEt-
Boc-Arg (Mts) -Gly-Asp (OBzl) -A recrystallized in hexane
3.55 g (2.7 mmol, yield 90%) of rg (Mts) -Gly-Asp (OBzl) was obtained as a colorless powder. Then Boc-Arg (Mts) -Gly-Asp
(OBzl) -Arg (Mts) -Gly-Asp (OBzl) 3.55 g (2.7 mmol) 1M-
It was dissolved in 100 ml of a solution of trifluoromethanesulfonic acid, thioanisole, m-cresol and trifluoroacetic acid, and stirred at room temperature for 1 hour. The reaction solution is poured into Et ₂ O to crystallize and Arg-
Gly-Asp-Arg-Gly-Asp 1.48 g (2.2 mmol, yield 81%) was obtained.

Synthesis Example 1 Synthesis of CM-chitin-RGDS (Compound 1) Viscosity 9 cps (1% solution, 20 ° C.), degree of etherification 0.
78 CM-chitin, CM with deacetylation degree of 0.5
-0.30 g of chitin (produced by Yaizu Suisan Chemical Co., Ltd.) at a pH of 7.
Dissolve in 4-phosphate buffer and keep water-soluble carbodiimide [1-ethyl-3-3- (dimethylaminopropyl) -carbodiimide] 128 mg 2.6m while keeping at 0 ° C.
1-phosphate buffer solution was added and the reaction was carried out for 1.5 hours. Then, the adhesive peptide Arg-Gly-Asp-Ser (RGD) dissolved in 8 ml of phosphate buffer.
S) (made by Kokusan Kagaku Kogyo Co., Ltd.) (400 mg) was added, and the mixture was reacted overnight at 4 ° C. The reaction solution was put in a Visking tube, dialyzed against ion-exchanged water and then pure water to remove low molecular weight components, and purified and lyophilized. Yield 0.24 g The structure was confirmed by IR and amino acid analysis. C
The structural formula of M-chitin RGDS is shown below.

[Chemical 1] Amino acid analysis (nmol / 10 μg) Arg: Gly: Asp: Ser = 2.06: 2.14: 1.99: 1.88 IR: Stretching vibration of amidocarbonyl (C═O) 16
52 cm ^-1

Synthesis Example 2 Succinylated CM-chitin-R
Synthesis of GDS (Compound 2) 20.0 g of CM-chitin of Synthesis Example 1 was dissolved in 100 ml of 1% triethylamine solution, and succinic anhydride 3
4.0 g and 4-dimethylaminopyridine 2.00 g were added and it stirred at room temperature all day and night. After completion of the reaction, the solution was poured into a large excess of acetone to reprecipitate succinylated CM-chitin. The precipitate was collected, washed with a large amount of methanol, washed with ether, and vacuum dried. Yield 22.40g.
Succinylated CM-chitin (0.30 g) was dissolved in a pH 7.4 phosphate buffer, and water-soluble carbodiimide (1-ethyl-3-3 (dimethylaminopropyl) -carbodiimide) 128 mg (2.6 ml) was added while keeping it at 0 ° C. An acid buffer solution was added and reacted for 1.5 hours. Then RGDS4 dissolved in 8 ml phosphate buffer
00 mg was added and reacted overnight at 4 ° C. The reaction solution
The mixture was placed in a Visking tube, dialyzed against ion-exchanged water and then pure water to remove low molecular weight components, and purified, and then freeze-dried. Yield 0.26 g The structure was confirmed by IR and amino acid analysis. The structural formula of succinylated CM-chitin-RGDS is shown below.

[Chemical 2] Amino acid analysis (nmol / 10 μg) Arg: Gly: Asp: Ser = 2.36: 2.13: 2.39: 2.00 IR: Stretching vibration of amidocarbonyl (C = O) 16
52 cm ^-1

Synthesis Example 3 Maleylated CM-chitin-RG
Synthesis of DS (Compound 3) 20.00 g of CM-chitin of Synthesis Example 1 and 36.6 g of maleic anhydride were reacted in the same manner as in Synthesis Example 2 to obtain 21.60 g of maleylated CM-chitin. 0.30 g of maleylated CM-chitin was dissolved in a pH 7.4 phosphate buffer, and the RGDS fragment was covalently bonded in the same manner as in Synthesis Example 2. Yield 0.33 g The structure was confirmed by IR and amino acid analysis. The structural formula of maleylated CM-chitin RGDS is shown below.

[Chemical 3] Amino acid analysis (nmol / 10 μg) Arg: Gly: Asp: Ser = 3.13: 2.78: 2.72: 2.57 IR: Stretching vibration of amidocarbonyl (C═O) 16
48 cm ^-1

Synthesis Example 4 Phtharoylated CM-chitin-R
Synthesis of GDS (Compound 4) 20.0 g of CM-chitin of Synthesis Example 1 and 50.0 g of phthalic anhydride were reacted in the same manner as in Synthesis Example 2 to produce phthaloylated CM.
22.31 g of chitin was obtained. 0.30 g of phthaloylated CM-chitin was dissolved in a pH 7.4 phosphate buffer,
The RGDS fragment was covalently bound in the same manner as in Synthesis Example 2. Yield 0.44 g The structure was confirmed by IR and amino acid analysis.
The structural formula of phthaloylated CM-chitin-RGDS is shown below.

[Chemical 4] Amino acid analysis (nmol / 10 μg) Arg: Gly: Asp: Ser = 2.30: 2.22: 1.89: 1.76 IR: Stretching vibration of amidocarbonyl (C═O) 16
52 cm ^-1

Synthesis Example 5 Itaconylated CM-chitin-R
Synthesis of GDS (Compound 5) 20.00 g of CM-chitin of Synthesis Example 1 and 38.0 g of itaconic anhydride were reacted in the same manner as in Synthesis Example 2 to obtain 21.45 g of itaconylated CM-chitin. Itaconylated CM-
Chitin (0.30 g) was dissolved in a pH 7.4 phosphate buffer, and the RGDS fragment was covalently bonded in the same manner as in Synthesis Example 2. Yield 0.36 g The structure was confirmed by IR and amino acid analysis. The structural formula of itaconylated CM-chitin-RGDS is shown below.

[Chemical 5] Amino acid analysis (nmol / 10 μg) Arg: Gly: Asp: Ser = 3.17: 3.25: 3.10: 2.88 IR: Stretching vibration of amidocarbonyl (C═O) 16
50 cm ^-1

Synthesis Example 6 trimellitylated CM-chitin-
Synthesis of RGDS (Compound 6) 20.00 g of CM-chitin of Synthesis Example 1 and 64.9 g of trimellitic anhydride were reacted in the same manner as in Synthesis Example 2 to obtain 23.74 g of trimellitylated CM-chitin. 0.30 g of trimellitylated CM-chitin was dissolved in a pH 7.4 phosphate buffer, and the RGDS fragment was covalently bonded in the same manner as in Synthesis Example 2. Yield 0.37 g The structure was confirmed by IR and amino acid analysis. The structural formula of trimellitylated CM-chitin-RGDS is shown below.

[Chemical 6] Amino acid analysis (nmol / 10 μg) Arg: Gly: Asp: Ser = 2.62: 2.79: 2.55: 2.27 IR: Stretching vibration of amidocarbonyl (C═O) 16
56 cm ^-1

Synthesis Example 7 CM-chitin-GRGD
Synthesis of S (Compound 7) Gly-Arg-G as an adhesive peptide fragment
CM-chitin-GRGDS was synthesized in the same manner as in Synthesis Example 1 using 460 mg of ly-Asp-Ser (GRGDS) (manufactured by Kokusan Kagaku Kogyo). Yield 0.36 g The structure was confirmed by IR and amino acid analysis. C
The structural formula of M-chitin-GRGDS is shown below.

[Chemical 7] Amino acid analysis (nmol / 10 μg) Arg: Gly: Asp: Ser = 1.53: 3.21: 1.34: 1.11 IR: Stretching vibration of amidocarbonyl (C = O) 16
54 cm ^-1

Synthesis Example 8 Succinylated CM-chitin-G
Synthesis of RGDS (Compound 8) GRGDS460m as an adhesive peptide fragment
Succinylated CM-chitin-GRGDS was synthesized in the same manner as in Synthesis Example 2 using g. Yield 0.39 g The structure was confirmed by IR and amino acid analysis. The structural formula of succinylated CM-chitin-GRGDS is shown below.

[Chemical 8] Amino acid analysis (nmol / 10 μg) Arg: Gly: Asp: Ser = 3.64: 7.06: 3.57: 3.30 IR: Stretching vibration of amidocarbonyl (C═O) 16
48 cm ^-1

Synthesis Example 9 Succinylated CM-chitin-R
Synthesis of GD (Compound 9) Using 460 mg of RGD as an adhesive peptide fragment, succinylated CM-chitin-RGD was synthesized in the same manner as in Synthesis Example 3. Yield 0.34 g The structure was confirmed by IR and amino acid analysis. The structural formula of succinylated CM-chitin-RGD is shown below.

[Chemical 9] Amino acid analysis (nmol / 10 μg) Arg: Gly: Asp: Ser = 4.46: 4.55: 4.49 IR: Stretching vibration of amidocarbonyl (C═O) 16
50 cm ^-1

Synthesis Example 10 Succinylated CM-chitin-
Synthesis of (RGD) ₂ (Compound 10) As an adhesive peptide fragment (RGD) ₂ 460 m
Succinylated CM-chitin- (RGD) ₂ was synthesized in the same manner as in Synthesis Example 4 using g. Yield 0.31 g The structure was confirmed by IR and amino acid analysis. The structural formula of succinylated CM-chitin- (RGD) ₂ is shown below.

[Chemical 10] Amino acid analysis (nmol / 10 μg) Arg: Gly: Asp = 6.05: 5.59: 5.90 IR: Stretching vibration of amidocarbonyl (C═O) 16
54 cm ^-1

Synthesis Example 11 Succinylated CM-chitin-
Synthesis of (RGD) ₃ (Compound 11) As an adhesive peptide fragment (RGD) ₃ 460 m
Succinylated CM-chitin- (RGD) ₃ was synthesized in the same manner as in Synthesis Example 5 using g. Yield 0.33 g The structure was confirmed by IR and amino acid analysis. The structural formula of succinylated CM-chitin- (RGD) ₃ is shown below.

[Chemical 11] Amino acid analysis (nmol / 10 μg) Arg: Gly: Asp = 6.69: 6.51: 6.23 IR: Stretching vibration of amidocarbonyl (C═O) 16
48 cm ^-1

Synthesis Example 12 Succinylated CM-chitin-
Synthesis of (RGD) ₅ (Compound 12) As an adhesive peptide fragment (RGD) ₅ 460 m
Succinylated CM-chitin- (RGD) ₅ was synthesized in the same manner as in Synthesis Example 6 using g. Yield 0.28 g The structure was confirmed by IR and amino acid analysis. The structural formula of succinylated CM-chitin- (RGD) ₅ is shown below.

[Chemical 12] Amino acid analysis (nmol / 10 μg) Arg: Gly: Asp: = 23.0: 21.1: 20.3 IR: Stretching vibration of amidocarbonyl (C═O) 16
56cm ^-1 stretching vibration of acid 1250,800Cm ^-1

Synthesis Example 13 Sulfated CM-chitin-RGD
Synthesis of S (Compound 13) C having the degree of etherification and deacetylation shown below
The method of M-chitin by Tokura et al. (Jpn. J. Cancer Res., 80,8
66-872 (1989), Cancer Res., 50, 3631-3637 (1990)), and sulfated CM-chitin having an etherification degree and a sulfation degree as shown in Table 1 was obtained.

Table 1 ────────────────────────────── Samples Etherification degree Sulfation degree Deacetylation degree ──── ────────────────────────── Compound 13-1 0.9 0.5 0.5 13-2 0.7 0.2 0 13-3 0.4 0.6 0 13-4 0.6 1.7 0 ────────────────────────────── Next, the RGDS fragment was covalently bonded in the same manner as in Synthesis Example 1 and Sulfated CM-chitin-R as shown
GDS was obtained. The structure was confirmed by IR and amino acid analysis. The structure of sulfated CM-chitin is shown below.
The amino acid analysis values of each sulfated CM-chitin-RGDS are shown in Table 2.

[Chemical 13]

[Chemical 14] Table 2 Amino acid analysis ^{* 1} ───────────────────────────── Sample Arg: Gly: Asp: Ser (nmol / 10μg) ── ─────────────────────────── Compound 13-1 2.89: 3.14: 3.19: 3.38 13-2 3.93: 4.27: 4.37: 3.81 13- 3 2.34: 2.30: 2.12: 1.42 13-4 4.30: 4.22: 3.43: 1.85 ───────────────────────────── * 1: Amino acid analysis Hydrolysis condition 6N-HCl, 10 hours, 110
° C IR: Stretching vibration of amidocarbonyl (C = O) 16
50 cm ^-1 Stretching vibration of sulfonic acid 1250, 800 cm ^-1

Synthesis Example 14 Sulfated CM-chitin-GRG
Synthesis of DS (Compound 14) Adhesive peptide fragment GRGDS (460 mg) and the sulfated CM-chitin of Synthesis Example 13 were covalently bonded in the same manner as in Synthesis Example 1 to synthesize sulfated CM-chitin-GRGDS. Yield 0.35 g The structure was confirmed by IR and amino acid analysis. The structural formula of sulfated CM-chitin-GRGDS is shown below.

[Chemical 15] Amino acid analysis (nmol / 10 μg) Arg: Gly: Asp: Ser = 2.51: 2.21: 2.42: 2.11 IR: Stretching vibration of amidocarbonyl (C = O) 16
55 cm ^-1

Synthesis Example 15 Synthesis of sulfated succinylated CM-chitin-RGDS (Compound 15) The succinylated CM-chitin of Synthesis Example 2 was sulfated in the same manner as in Synthesis Example 13 and treated in the same manner as in Synthesis Example 2 to prepare an RGDS fragment. Was covalently bonded. Yield 0.37 g The structure was confirmed by IR and amino acid analysis. The structural formula of sulfated succinylated CM-chitin-RGDS is shown below.

[Chemical 16] Amino acid analysis (nmol / 10 μg) Arg: Gly: Asp: Ser = 2.61: 2.77: 2.59: 2.22 IR: Stretching vibration of amidocarbonyl (C═O) 16
54 cm ^-1

Synthetic Example 16 Synthesis of sulfated succinylated CM-chitin-GRGDS (Compound 16) The succinylated CM-chitin of Synthetic Example 2 was sulfated in the same manner as in Synthetic Example 13, and the GRGDS fragment was synthesized in the same manner as in Synthetic Example 7. Was covalently bonded. Yield 0.31 g The structure was confirmed by IR and amino acid analysis. The structural formula of sulfated succinylated CM-chitin-GRGDS is shown below.

[Chemical 17] Amino acid analysis (nmol / 10 μg) Arg: Gly: Asp: Ser = 3.84: 7.50: 3.68: 3.26 IR: Stretching vibration of amidocarbonyl (C═O) 16
52 cm ^-1

Synthesis Example 17 CM-chitin-RGDSG-
Synthesis of NH ₂ (compound 17) Arg-Gly-Asp-Ser-Gly-NH ₂ (RGDSG-N) with an amidated carboxyl terminus as an adhesive peptide fragment
CM-chitin-RGDSG-NH ₂ was synthesized in the same manner as in Synthesis Example 1 using 460 mg of H ₂ ). Yield 0.36
Confirmation of the g structure was carried out by IR and amino acid analysis. C
Of M- Chitin -RGDSG-NH ₂ The structural formula is shown below.

[Chemical 18] Amino acid analysis (nmol / 10 μg) Arg: Gly: Asp: Ser = 1.82: 3.82: 1.84: 1.61 IR: Stretching vibration of amidocarbonyl (C = O) 16
58 cm ^-1

Synthesis Example 18 Synthesis of CM-chitin-RGDS (Compound 18) RGDS 1.5 g as an adhesive peptide fragment
And CM-chitin-RGDS in the same manner as in Synthesis Example 1.
Was synthesized. Yield 0.32 g The structure was confirmed by IR and amino acid analysis. C
The structural formula of M-chitin-RGDS is shown below.

[Chemical 19] Amino acid analysis (nmol / 10 μg) Arg: Gly: Asp: Ser = 4.10: 4.27: 3.98: 3.78 IR: Stretching vibration of amidocarbonyl (C═O) 165
5 cm ^-1

Synthesis Example 19 CM-chitin-GRGDSP
Synthesis of (Compound 19) Adhesive peptide fragment-6 (Production Example 3) Gly
-Arg-Gly-Asp-Ser-Pro (GRGD
SP) using 480 mg and in the same manner as in Synthesis Example 1 CM-
Chitin-GRGDSP was synthesized. Yield 0.35 g The structure was confirmed by IR and amino acid analysis. C
The structural formula of M-chitin GRGDSP is shown below.

[Chemical 20] Amino acid analysis (nmol / 50 μg) Arg: Gly: Asp: Ser: Pro = 1.53: 3.21: 1.34: 1.11: 1.33 IR: Stretching vibration of amidocarbonyl (C = O) 16
57 cm ^-1

Test Example 1 (Experimental lung metastasis) The compounds of the present invention were examined for their cancer metastasis inhibitory action.
RGDS-carrying CM-chitin derivatives (Compounds 1 to 10) were added as 1000 μg each as a highly metastatic cancer cell B
Each 16-BL6 melanoma cell was mixed in 0.2 ml of PBS, and 0.2 ml of the mixture was intravenously injected into 5 female C57BL / 6 mice per group. The injected mixture (0.2 ml) contained 5 × 10 ⁴ B16-BL6 cells. Fourteen days after the administration, the number of mouse lung colonies was counted and compared with the control PBS administration group, and the metastasis inhibition rate was calculated by the following formula. Metastasis inhibition rate (%) = (1- (Number of lung colonies in the sample administration group / PBS
The number of lung colonies in the administration group)) × 100 The results are shown in Table 3. Table 3 ──────────────────────── Sample metastasis inhibition rate (range) ────────────────── ─────── Compound 1 98% (100-93%) Compound 2 77% (85-68%) Compound 3 73% (88-66%) Compound 4 65% (83-42%) Compound 5 56 % (67-44%) Compound 6 88% (92-75%) Compound 7 68% (83-51%) Compound 8 47% (52-40%) Compound 9 66% (73-38%) Compound 10 80 % (91-70%) Compound 11 66% (78-53%) Compound 12 80% (89-66%) Compound 13-1 54% (64-27%) Compound 13-2 50% (86-16%) ) Compound 13-3 52% (82-23%) Compound 13-4 23% (42-6%) Compound 14 50% (72-44%) Compound 15 68% (76-60%) Compound 16 73% ( 80-63%) Compound 17 56% (65-39%) Compound 18 49% (67-40%) Compound 19 51% (73-37%) Comparative sample RGDS (Arg-Gly-Asp-Ser) 8% ( 38-(-32)%) ────────────────────────

Even when the compound of the present invention was not mixed with B16-BL 6 cells and was intravenously administered to mice 5 minutes after the administration of B16-BL 6 cells, a high metastasis-inhibiting effect was still obtained. ..
This result indicates that the compound of the present invention can be administered by an appropriate method such as intravenous injection to obtain a cancer metastasis suppressing effect. In addition, when the compound group of the present invention was subjected to a toxicity test using mice, no toxicity was observed at all.

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Hiroyuki Komazawa, Inventor, 210 Nakanuma, Minamiashigara City, Kanagawa Prefecture, Fuji Photo Film Co., Ltd. (72) Inventor Ichiro Higashi 5-3-2 Makomanaikamimachi, Minami-ku, Sapporo-shi, Hokkaido

Claims (3)

[Claims] 1. An arginine residue-glycine residue-on the side chain
A cancer metastasis inhibitor comprising a carboxymethylated chitin derivative having a peptide sequence consisting of an aspartic acid residue or a pharmacologically acceptable salt thereof as an active ingredient. 2. A carboxymethylated chitin derivative having an adhesive peptide represented by the following general formula [1] bonded to a side chain via an amide bond, an ester bond, an ether bond or a urethane bond or a derivative thereof. The cancer metastasis inhibitor according to claim 1, which comprises a pharmacologically acceptable salt as an active ingredient. [R ¹ ]-[CO]-([X] -Arg-Gly-Asp- [Y]) n- [Z]-[R ² ] [1] In the formula, Arg is arginine, Gly is glycine, and Asp is Indicates an aspartic acid residue. [] Represents that the group in [] may or may not be present. When present, X and Y are amino acid residues selected from serine (Ser), glycine (Gly), valine (Val), asparagine (Asn), aspartic acid (Asp), proline (Pro), or amino acids thereof. A peptide residue composed of residues is shown, and Z is -O- or -NH-. One of R ¹ and R ² is hydrogen, or a linear or branched alkyl group having 1 to 9 carbon atoms or a carbon which may have a substituent and may contain an unsaturated bond. The number represents an aryl group having 6 to 9, and the other is a linear or branched alkylene group having 1 to 9 carbon atoms or a carbon which may have a substituent and may include an unsaturated bond. It represents an arylene group having a number of 6 to 9. n is 1
Indicates an integer of 5. 3. A molecular weight of about 3,000 to 100,000.
3. The cancer metastasis inhibitor according to claim 1 or 2, which comprises a carboxymethylated chitin derivative or a pharmacologically acceptable salt thereof within the range of 10 as an active ingredient.