Self-magnetic metal-salen complex compound

Abstract

A new self-magnetic metal-salen complex compound and its derivatives are provided. The present invention is a metal-salen complex compound including any one of the following compounds. ##STR00001##

Claims

1. An antitumor drug containing a magnetic drug containing a self-magnetic metal-salen complex compound represented by a formula (I), wherein the formula (I) is a following compound (II): ##STR00021## wherein M is Fe, and each of a-f is a methyl group, or each of the a-f is a hydrogen, or each of b and e is a hydrogen and a, c, d, and f is a methyl group, and wherein the magnetic drug being guided to a target tissue by administering the magnetic drug into a body and then irradiating the magnetic drug with an external magnetic field.

2. The antitumor drug of claim 1, wherein each of the a-f is hydrogen.

3. The antitumor drug of claim 1, wherein each of the a-f is a methyl group.

4. The antitumor drug of claim 1, wherein each of the a, c, d and f is a methyl group.

5. The antitumor drug of claim 1, wherein a charge transfer of a side chain of the metal-salen complex compound of the formula (I) is less than 0.5 electrons (e).

6. The antitumor drug containing the magnetic drug of claim 1, as an active ingredient.

7. A magnetic drug guiding system comprising: a means for supplying a magnetic field to the drug stated in claim 1, and introduced into a body and a means for moving the magnetic field to an affected site.

8. A method of treating tumors using the antitumor drug according to claim 1, wherein the method comprises administering the antitumor drug into a body, guiding the antitumor drug to a target issue including tumors by applying an external magnetic field and diminishing tumor expansion.

9. The antitumor drug of claim 2, wherein a charge transfer of a side chain of the metal-salen complex compound of the formula (I) is less than 0.5 electrons (e).

10. The antitumor drug of claim 3, wherein a charge transfer of a side chain of the metal-salen complex compound of the formula (I) is less than 0.5 electrons (e).

11. The antitumor drug of claim 4, wherein a charge transfer of a side chain of the metal-salen complex compound of the formula (I) is less than 0.5 electrons (e).

12. The antitumor drug containing the magnetic drug of claim 2, as an active ingredient.

13. The antitumor drug containing the magnetic drug of claim 3, as an active ingredient.

14. The antitumor drug containing the magnetic drug of claim 4, as an active ingredient.

15. A magnetic drug guiding system comprising a means for supplying a magnetic field to the drug stated in claim 2, and introduced into a body and a means for moving the magnetic field to an affected site.

16. A magnetic drug guiding system comprising a means for supplying a magnetic field to the drug stated in claim 3, and introduced into a body and a means for moving the magnetic field to an affected site.

17. A magnetic drug guiding system comprising: a means for supplying a magnetic field to the drug stated in claim 4, and introduced into a body and a means for moving the magnetic field to an affected site.

18. A method of treating tumors using the antitumor drug according to claim 2, wherein the method comprises administering the antitumor drug into a body, guiding the antitumor drug to a target issue including tumors by applying an external magnetic field and diminishing tumor expansion.

19. A method of treating tumors using the antitumor drug according to claim 3, wherein the method comprises administering the antitumor drug into a body, guiding the antitumor drug to a target issue including tumors by applying an external magnetic field and diminishing tumor expansion.

20. A method of treating tumors using the antitumor drug according to claim 4, wherein the method comprises administering the antitumor drug into a body, guiding the antitumor drug to a target issue including tumors by applying an external magnetic field and diminishing tumor expansion.

Description

BRIEF DESCRIPTION OF DRAWINGS

(1) FIG. 1 is a diagrammatic illustration of a state where a bar magnet is made to be in contact with a rectangular flask containing a culture medium of rat L6 cells.

(2) FIG. 2 is a graph showing the results of calculation of the number of cells as a result of taking photographs of a bottom face of the rectangular flask from its one end to the other end.

(3) FIG. 3 is a perspective view of a magnetic guide device.

(4) FIG. 4 is a graph showing MRI measurement results of kidneys to which a magnetic field was applied after intravenously injecting a metal-salen complex to mice.

(5) FIG. 5 is photographs of mouse tail tendon tissues showing the effects of the salen complex on melanoma growth in mice.

(6) FIG. 6 is a graph showing the effects of the salen complex on melanoma growth in mice.

(7) FIG. 7 is microphotographs showing the results of a histological examination indicating the effects of the salen complex on melanoma growth in mice.

DESCRIPTION OF EMBODIMENTS

(8) Preferred embodiments of the self-magnetic metal-salen complex compound represented by Formula (I) are (II) to (XI) below.

(9) (II)

(10) X, Y: six-membered ring structure

(11) (a to h)=H

(12) ##STR00005##
(III)

(13) X, Y: six-membered ring structure

(14) (c, f)=C(O)H

(15) (a, b, d, e, g, h)=H

(16) ##STR00006##
(IV)

(17) X, Y: five-membered ring structure, (a, c, d, e, f, h)=H

(18) ##STR00007##
(V)

(19) X, Y: six-membered ring structure

(20) (a, b, g, h): H

(21) (e, f), (g, h): constitute part of furan and furan is condensed with a main skeleton.

(22) M: Fe

(23) ##STR00008##
(VI)

(24) X, Y: six-membered ring structure

(25) (a, h): constitute part of cyclohexane and cyclohexane is condensed with a main skeleton.

(26) (c, d), (e, f): constitute benzene

(27) (b, g): H

(28) M: Fe

(29) ##STR00009##
(VII)

(30) X, Y: six-membered ring structure

(31) (a, h): constitute part of benzene

(32) (c, d), (e, f): constitute benzene

(33) (b, g): H

(34) M: Fe

(35) ##STR00010##
(VIII)

(36) X, Y: six-membered ring structure

(37) (c, d), (e, f): constitute anthracene

(38) (a, b, g, h): H

(39) M: Fe

(40) ##STR00011##
(IX)

(41) X, Y: six-membered ring structure

(42) (c, d), (e, f): constitute anthracene

(43) (a, b, g, h)=H

(44) Isomer of (V)

(45) M: Fe

(46) ##STR00012##
(X)

(47) X, Y: six-membered ring structure

(48) (c, d), (e, f): constitute benzene

(49) Side chains at meta positions of benzene are halogens (bromine).

(50) (a, b, g, h): H

(51) M: Fe

(52) ##STR00013##
(XI)

(53) X, Y: six-membered ring structure

(54) (c, d), (e, f): constitute benzene

(55) Side chains at meta positions of benzene are methoxyl groups.

(56) (a, b, g, h): H

(57) M: Fe

(58) ##STR00014##

(59) Since the metal-salen complex compound of Formula (I) has magnetic properties so that it can be guided by an external magnetic field, an absolute value of charge transfer of its side chain should preferably be less than 0.5 electrons (e). The substituted compound described in, for example, WO 2010/058280 can be used as the aforementioned R.sub.3 which constitutes the above-described side chain. The content of the above-mentioned publication constitutes descriptions of the specification of this application.

(60) As the metal-salen complex compound represented by Formula (I) is administered to inside the body and then guided to the target tissue by irradiating the body with the external magnetic field, the magnetic force strength of the metal-salen complex compound and the magnetization strength of the drug containing this metal-salen complex compound as an active ingredient are respectively within the range from 0.5 to 1.5 emu/g. The magnetic drugs are mainly injections or transfusions, or may be powdered drugs. Saline can be preferably used as a solvent for the injections or the transfusions. The magnetic drug contain the metal-salen complex as an active ingredient, for example, by 50 wt % or more; and in addition, the magnetic drug may contain diluents, stabilizing agents, and second active pharmaceutical ingredients which will have no or little influence on effectiveness, physical properties, or chemical properties of the metal-salen complex. The aforementioned metal-salen complex compound can be used as an anticancer agent.

(61) Examples of the means for supplying a magnetic field to the magnetic drug after applying the magnetic drug into a human or animal body are permanent magnets or induction magnetic fields such as MRI. The strength of the external magnetic field should preferably be within the range of 0.5 to 1.0 T, or more preferably 0.8 to 1.0 T. Examples of the means for moving the magnetic field to the affected site include MRI besides an X-Y table for moving the permanent magnet. Examples of a form for supplying the magnetic field to the affected site tissues include a form for supplying the magnetic field from the body surface and a form for setting the magnetic field generating means at blood vessels near the affected site tissues. In order to supply the magnetic field from the body surface, there is a form for supplying the magnetic field from the front of the body and/or the back of the body.

EXAMPLES

(62) Next, examples of the present invention will be explained.

Example 1

Synthesis of Metal-Salen Complex Compound (II)

First Synthesis Example

(63) The metal-salen complex compound (II) was synthesized in accordance with the following reaction formulas.

(64) ##STR00015##

Synthesis of Compound 1

(65) P-TsOH (10 mg) was added to an ethyl formate solution (60 ml) containing glycine methyl ester monohydrochloride (10.0 g, 0.079 mol) and the obtained solution was heated to boiling. Several drops of triethylamine were put into the solution while being boiled; and the mixed solution was brought to reflux for 24 hours and then cooled down to room temperature. Subsequently, white triethylaminehydrochloride was filtered and the residue was concentrated to 20 ml. The obtained solution was cooled down to a temperature of 5 degrees Celsius and then filtered. A reddish brown concentrated solution which was a residue (Compound 1) was obtained.

Synthesis of Compound 2

(66) CH.sub.2Cl.sub.2 (20 ml) was dissolved in Compound 1. Then, ethane-1,2-diamine (1.2 g) and acetic acid (HOAc) (20 l) were added to the obtained solution; and this reacted mixed solution was then brought to reflux for 6 hours. Subsequently, the reactant mixed solution was cooled down to room temperature, thereby obtaining 4 g of a yellow oil concentrate (Compound 2). Purity of the obtained Compound 2 was enhanced by flash column chromatography by using silica gel.

Synthesis of Compound 0

(67) Compound 2 and triethylamine were introduced in methanol (50 ml) and a solution of metallic chloride (FeCl.sub.3(4H.sub.2O) when synthesizing the iron-salen complex compound) was mixed in methanol (10 ml) in a nitrogen atmosphere. The mixture were mixed for one hour in a nitrogen atmosphere, thereby obtaining a brown compound. Then, this compound was dried in a vacuum, the obtained compound was diluted with dichloromethane (400 ml), washed twice with a saline solution, was dried over Na.sub.2SO.sub.4, and then dried in a vacuum, thereby obtaining Compound 0 (the metal-salen complex compound (II)).

Second Synthesis Example

(68) The metal-salen complex compound (II) was synthesized in accordance with the following reaction formulas.

(69) ##STR00016##

(70) Compound 5 was synthesized by introducing 3.4 g of 3-methylacetylacetone (Compound 4) and 0.9 g of ethylene diamine (Compound 3) into anhydrous methanol (50 ml) while adjusting pH to pH 6 by using acetic acid on ice. The obtained solution was brought to reflux for 15 minutes and allowed to evaporate until its volume reduces to one half its original volume. Then, water of the same volume was added to the solution and let it deposit, thereby obtaining 1.4 g of white compound (Compound 5).

(71) Subsequently, Compound 5 (1.2 g, 5 mmol) was introduced into methanol (50 ml) and FeSO.sub.4.7H.sub.2O (1.4 g, 5 mmol) was added to the obtained solution, thereby obtaining a pale bluish green solution. As this mixed solution was stirred for 8 hours at room temperature in a nitrogen atmosphere, its color gradually changed to brown. Subsequently, the solution was allowed to evaporate to reduce a half of its volume and then the same volume of water was added to the obtained solution. Next, vacuum was produced to allow methanol to evaporate, thereby obtaining brown lumps. These lumps were gathered, washed with water, and dried by producing a vacuum, thereby obtaining 360 mg of the target compound (the metal-salen complex compound (II)).

Third Synthesis Example

(72) The metal-salen complex compound (II) was synthesized in accordance with the following reaction formulas.

(73) ##STR00017##

(74) Iron (II) acetate (0.83 g, 4.8 mmol) and degassed methanol (48 ml) were introduced into a reaction container in a nitrogen atmosphere and acetylacetone (0.95 g, 9.5 mmol) was added to the obtained solution. The solution was stirred in a reflux for 15 minutes and then let it stand to cool. Deposited crystals were filtered and the obtained solution was washed with cooled methanol (10 ml). Subsequently, the solution was dried under reduced pressure, thereby obtaining 1.07 g of an intermediate.

(75) The intermediate (1.07 g, 3.4 mmol), ligand atoms (0.70 g, 3.4 mmol), and degassed decalin (30 ml) were introduced into a reaction container in a nitrogen atmosphere and the obtained solution was stirred in a reflux for 1 hour. After letting the solution stand to cool and filtering a deposited solid, the obtained solid was washed with degassed cyclohexane (10 ml). Next, the solution was dried under reduced pressure, thereby obtaining 0.17 g of a product (the metal-salen complex compound (II)).

Example 2

Synthesis of Metal-Salen Complex Compound (III)

(76) The metal-salen complex compound (III) was synthesized in accordance with the following reaction formulas.

(77) ##STR00018##

(78) Iron (II) acetate (0.78 g, 4.5 mmol) and degassed methanol (20 ml) were introduced into a reaction container in a nitrogen atmosphere and acetylacetone (0.91 g, 9.9 mmol) was added to the obtained solution. The solution was stirred in a reflux for 15 minutes and then let it stand to cool. Deposited crystals were filtered, and the obtained solution was washed with cooled methanol (10 ml). Subsequently, the solution was dried under reduced pressure, thereby obtaining 0.58 g (yield 67%) of an intermediate.

(79) The intermediate (240 mg, 0.75 mmol), ligand atoms (210 mg, 0.75 mmol), and degassed decalin (10 ml) were introduced into a reaction container in a nitrogen atmosphere and the obtained solution was stirred in a reflux for 30 minutes. After letting the solution stand to cool and filtering a deposited solid, the obtained solid was washed with degassed cyclohexane (3 ml). Next, the solution was dried under reduced pressure, thereby obtaining 101 mg of a product (the metal-salen complex compound (III)).

Example 3

Synthesis of Metal-Salen Complex Compound (IV)

(80) The metal-salen complex compound (IV) was synthesized in accordance with the following reaction formulas.

(81) ##STR00019##

(82) Iron (II) acetate (0.83 g, 4.8 mmol) and degassed methanol (48 ml) were introduced into a reaction container in a nitrogen atmosphere and acetylacetone (0.95 g, 9.5 mmol) was added to the obtained solution. The solution was stirred in a reflux for 15 minutes and then let it stand to cool. Next, Compound 7 (120 mg, 2.0 mmol) and SiO.sub.2 (1 g) were added to a solution of Compound 6 (60 mg, 1.0 mmol) dissolved in CH.sub.2Cl.sub.2 (10 ml); and the obtained solution was stirred all night at room temperature to cause a reaction, thereby synthesizing Compound 8. Subsequently, the obtained Compound 8 together with iron (II) acetate (0.83 g, 4.8 mmol) and degassed methanol (48 ml) was introduced into a reaction container in a nitrogen atmosphere and acetylacetone (0.95 g, 9.5 mmol) was added to the obtained solution. The solution was stirred for 15 minutes in a reflux and deposited crystals were filtered, thereby obtaining a brown target compound (the metal-salen complex compound (IV)).

Example 4

(83) The compounds (V) to (XI) were synthesized by a method described on pages 43 to 47 of a specification of WO2010/058280. Bromine or a methoxyl group which is a side chain is added to a main skeleton, when forming a metal complex bond to salen, by substituting a protecting group (NHBoc), which is bonded to a benzene ring at a para position with an OH group of the benzene ring, with bromine or the methoxyl group. With the compounds of (VIII) and (IX) in which (c, d) and (e, f) constitute anthracene, the following compound is used as a starting material instead of para-nitrophenol.

(84) ##STR00020##

(85) Regarding synthesis of the metal-salen complex (VI) in which (a, h) constitute cyclohexane and the metal-salen complex (VII) in which (a, h) constitute benzene, the target salen before forming a coordinate bond with a metal is produced by a method described in Journal of Thermal Analysis and calorimetry, Vol. 75 (2004) 599-606, Experimental on page 600.

Example 5

(86) Whether the aqueous solution of each metal-salen complex (II) to (XI) could be trapped by the permanent magnet or not was examined while having a pump circulate the aqueous solution in a glass tube. A circulation speed of the aqueous solution of the metal-salen complex was 100 mm/s, the diameter of the glass tube was 1.3 mm, the distance between the surface of the glass tube and the permanent magnet was 1.35 mm, and a concentration of the compound was 10 mg/ml. The magnet used was a commercially available bar magnet with a circular cross-section (20 mm in diameter; 150 mm long; model number N50 by Shin-Etsu Chemical Co., Ltd.; maximum magnetic flux density 0.8 T). Each metal complex was checked if it was trapped in an area where it traps on the magnet.

Example 6

(87) Regarding each of the metal (iron)-salen complexes (II) to (XI) obtained by the above-described methods, powder of the iron-salen complex compound (10 mg) was applied, to the degree allowing its magnetic attraction to be visibly observed, to a culture medium (PBS) when the rat L6 cells were in a 30% confluent state; and the state of the culture medium was photographed after 48 hours. FIG. 1 is a diagrammatic illustration of a state in which a bar magnet is in contact with a rectangular flask containing the rat L6 cell culture medium. After 48 hours, an image of the bottom of the rectangular flask was photographed from one end to the other end and the number of cells was calculated and the results are shown in FIG. 2. Referring to FIG. 2, a proximal position from the magnet indicates within a project area of the magnet end face on the bottom of the square-type flask and a distal position from the magnet indicates an area on the opposite side of the magnet end face on the bottom of the rectangular flask

(88) FIG. 2 shows that a concentration of each metal-salen complex increases as the metal-salen complex is attracted at the proximal position from the magnet; and it can be seen that the number of cells becomes extremely lower than that at the distal position due to a DNA-growth inhibition action of the complex. As a result, the drug can be concentrated at the target affected site or tissues of the individual by means of the metal-salen complex and the system including a magnetism-generating means.

(89) Next, a guide example using a guide device will be explained. With this guide device, as illustrated in FIG. 3, a pair of magnets 230 and 232 facing each other in the direction of gravity are supported by a stand 234 and a clamp 235, and a metal plate 236 is located between the magnets 230 and 232. A magnetic field of uniform strength can be created locally by placing the metal plate, especially an iron plate, between the pair of magnets. An electrical magnet can be used instead of the magnet to modify the magnetic force generated in this guide device. The magnetism-generating means can be moved to a target position of the individual on a table to allow the pair of magnetism-generating means to move in X, Y, and Z directions.

(90) The drug can be concentrated on a specific tissue by placing a tissue in the region of the magnetic field. After intravenously injecting the aforementioned metal-salen complex (drug concentration: 5 mg/ml (15 mmol)) to a mouse weighing about 30 g, a laparotomy was performed, and the mouse was placed on the iron plate to locate its right kidney between the pair of magnets. The magnets used were Product No. N50 (neodymium permanent magnets) by Shin-Etsu Chemical Co., Ltd. with a residual flux density of 1.39 to 1.44 T. Under this circumstance, the magnetic field applied to the right kidney was about 0.3 T, and the magnetic field applied to its left kidney was about 1/10 of the above-mentioned magnetic field.

(91) Together with the left kidney and a kidney to which no field was applied (Control), a magnetic field was applied to the right kidney of the mouse; and after 10 minutes the SNR was measured by MRI in T1 mode and T2 mode. As a result as shown in FIG. 4, it was confirmed that the drug stayed in the right kidney (RT) to which the magnetic field was applied, as compared to the left kidney (LT) and Control.

(92) FIG. 5 shows the effect of the salen complex on melanoma growth in mice. Melanoma was established in mouse tail tendons in vivo by local grafting of cultured melanoma cells (Clone M3 melanoma cells). The salen complex (50 mg/kg) was administered intravenously via tail tendon vein, followed by local application of a magnetic field by using a commercially available bar magnet (630 mT, a cylindrical neodymium magnet, 150 mm long and 20 mm in diameter). Application of a bar magnet was performed with 3 hour gentle contact with the site of melanoma immediately after injection of the salen complex for 10-14 days.

(93) Application of the bar magnet was performed in such a way so that the magnetic field strength became maximal over the area of expected melanoma extension, which was approximately 150 mm or shorter in mouse tail tendons with the growth period of 2 weeks. Twelve days after the initial injection of the metal-salen complex, the extension of melanoma was evaluated by assessing the area of melanoma pigmentation. As shown in FIG. 6, the melanoma extension was greatest in the saline group (10017.2%), in which saline was injected instead of the metal-salen complex.

(94) Meanwhile, the melanoma extension modestly decreased (63.6816.3%) in the SC group, in which the salen complex was injected without the application of a magnetic force field. In contrast, most melanoma disappeared (9.053.42%) in the SC+Mag group, in which the metal-salen complex was injected while applying a magnetic field.

(95) A histological examination was performed as shown in FIG. 7 by means of Hematoxylin-Eosin staining (HE) and immunohistological staining (Ki, CyclinD1) with an anti-Ki-67 antibody and an anti-Cyclyn D1 antibody which are tumor proliferation markers in tissue sections. As a result, the histological examination revealed that tumor expansion of melanoma which was prominent before administration diminished when the metal-salen complex was injected (SC); and the tumor expansion of melanoma mostly disappeared after the application of the salen complex when the magnetic field application was combined with administration of the salen complex (as indicated as an ellipse in a dotted line in FIG. 7).