Technical Field This invention relates to cyclic etheramine derivatives useful as remedies for malignant tumors, the prognoses of which are not promising, such as neuroblastomas.

Background Art Among infantile malignant tumors, progressive neuroblastoma (NB) is still not promising in prognosis in spite of multidisciplinary treatment. From a study conducted using <BR> 13-cis-retinoic acid, it is known that a down regulation takes place in the expression of the N-myc gene to exhibit differentiation-inducing effect. This acid is, however, not considered to bring about sufficient clinical effects for neuroblastomas.

An object of the present invention is, therefore, to provide a medicament effective for malignant tumors, the prognoses of which are not promising, such as neuroblastomas.

Disclosure of the Invention By stimulating cultured cell strains of human neuroblastoma origin with nerve growth factor (NGF), the

present inventors conducted an investigation as to whether or not the expression of c-fos, which is one of the initial <BR> <BR> expression gene groups located downstream of theintracellular signal transduction system, would take place. As a result, cultured cell strains-in each of which upon NGF stimulation, the signal was transducted to c-fos-were found to be those recognized to express NGF high affinity receptor (trk-A) and <BR> <BR> NGFlowaffinityreceptor (LNGFR). Itwasalsosuggestedthat each cultured cell strain, on which the expression of c-fos was not confirmed despite the possession of both of the receptors, developed a certain abnormality in the course of the transduction of the signal.

Therefore, the present inventors synthesized a variety of compounds, and conducted an investigation as to whether or not they would repair the signal transduction system in the case of the above-described abnormality in the expression of c-fos and would act to induce differentiation. As a result, it was found that cyclic etheramine derivatives represented by the below-described formula have effect to repair the expression of c-fos. The present inventors proceeded with a further investigation on the c-fos expression reparative effect of these compounds. As a result, it has been found that these compounds also have effect to maintain the c-fos expression reparative effect for a long time and further, to increase the expression of c-fos, leading to the completion of the present invention.

Described specifically, the present invention provides a cyclic etheramine derivative represented by the following formula (1) :

(1) whereinYlandY2maybethesameordifferentandeachrepresents an oxygen atom or two hydrogen atoms, and Ri and R2 may be the same or different and each represents a hydrogen atom or a substituted or unsubstituted alkyl group ; or a salt thereof.

The present invention also provides a medicament which comprises as an active ingredient the cyclic etheramine derivative represented by the formula (1) or the salt thereof.

The present invention further provides a medicament composition which comprises the cyclic etheramine derivative represented by the formula (1) or the salt thereof and a pharmaceutically acceptable carrier.

The present invention further provides use of the cyclic etheramine derivative represented by the formula (1) or the salt thereof for the production of the medicament.

The present invention still further provides a method for treating a malignant tumor, which comprises administering an effective amount of the cyclic etheramine derivative represented by the formula (1) or the salt thereof.

Best Modes for Carrying Out the Invention In the formula (1), Y'and y2 may be the same or dif f erent and each represents an oxygen atom or two hydrogen atoms. In this definition, the representation of two hydrogen atoms by each of Y1 and Y2 is more preferred. <BR> <BR> <BR> <BR> <P> R1andR2maybethesameordifferentandeachrepresents<BR> <BR> <BR> <BR> <BR> <BR> a hydrogen atom or a substituted or unsubstituted alkyl group.

In this definition, the alkyl group may preferably be a C1-C8 alkyl group, with a C1-C6 alkyl group being more preferred.

The alkyl group can be linear or branched. Examples of the alkyl group can include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, n-pentyl and n-hexyl.

Illustrative of substituents which can substitute the alkyl group are carboxyl groups, alkoxycarbonyl groups, hydroxyl groups, halogen atoms, substituted or unsubstituted aryl groups, and substituted or unsubstituted heterocyclic groups. Examples of the alkoxycarbonyl groups can include C1-C6 alkoxycarbonyl groups, more specifically methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl and butoxycarbonyl. Examples of the halogen atoms can include

fluorine atoms, chlorine atoms, bromine atoms and iodine atoms.

Examples of the aryl groups can include C6-C1o aryl groups, more specifically phenyl and naphthyl. Examples of the heterocyclic group can include 5-or 6-membered, nitrogen-containing heterocyclic groups, more specifically pyrrolyl, imidazolyl, pyrazolyl, pyridyl, pyrimidinyl and pyrazinyl. Of these, pyridyl is particularly preferred. It is to be noted that this pyridyl group can include such a case that the nitrogen atom in the pyridine ring is bonded with an alkyl group to form a pyridinium salt.

Illustrative of groups which can substitute on these aryl and heterocyclic groups are an amino group, C1-C6 alkylamino groups and di (C1-C6 alkyl) amino groups. More specific substituents can include amino, methylamino, ethylamino, propylamino, dimethylamino, diethylamino and diisopropylamino. Of these, di (C1-C6alkyl) amino groups are <BR> <BR> <BR> <BR> <BR> morepreferred, withadimethylaminogroupbeingparticularly preferred.

Preferred examples of the salt of the cyclic etheramine derivative represented by the formula (1) are mineral salts <BR> <BR> <BR> <BR> suchasthehydrochloride, sulfateandnitrate. WhenR1and/or R2 contain a group with a pyridinium salt formed therein, its counter ion may preferably be a halogen ion such as a chlorine ion or a bromine ion.

In the present invention, solvates represented by

hydrates are also included. The compound according to the present invention can have one or more asymmetric carbon atoms depending on the substituents. In such a case, optically active isomers are also included in the present invention.

The cyclic etheramine derivative represented by the formula (1) or the salt thereof, which may hereinafter be referred to as"the compound (1) of the present invention", can be produced, for example, in accordance with the following reaction scheme:

wherein X1 and X2 each independently represents a halogen atom,

R3 represents a hydrogen atom, an alkyl group, a phenyl group or a substituted or unsubstituted phenyl group, R4 represents <BR> <BR> <BR> <BR> <BR> analkylgroup, R5representsanalkylgroup, andR6represents a substituted or unsubstituted aryl group or a substituted or unsubstituted heterocyclic group.

Specifically described, a halogenoacetic acid ester (3) is reacted with 4, 4'-dihydroxydiphenylmethane (2) to afford a compound (4). Ester residual groups of the compound (4) <BR> <BR> <BR> <BR> <BR> may be converted into active ester moieties as needed, followed by the reaction with ammonia to yield a compound (5). The compound (5) is then reduced to obtain the compound (6). The compound (6) and the compound (4) are reacted to afford a cyclic etheramine (la). Reduction of this compound (la) provides <BR> <BR> <BR> <BR> <BR> acompound (lb), withwhichana, ß-unsaturatedfattyacidester or halogenofatty acid ester is reacted to obtain a compound (lc). Reduction of this compound (1c) provides a compound (1d), halogenation of which results in a compound (le).

Reaction of desired one of various aryl compounds and heterocyclic compounds with the compound (le) provides a compound (1f).

A description will hereinafter be made reaction step by reaction step.

Step (a) In the step (a), the halogenoacetic acid ester is reacted with 4, 4'dihydroxydiphenylmethane (2) to obtain the compound

(4).

Illustrative of the halogenoacetic acid ester are methyl bromoacetate and ethyl bromoacetate. This reaction can be conducted prefer ably in the presence of abase such as potassium carbonate, sodium hydroxide or sodium carbonate. It is also preferred to conduct this reaction at 0 to 100°C in a polar solvent such as dimethylformamide.

The ester residual groups of the compound (4) may be converted into active ester moieties, for example, substituted aryl ester moieties typified, for example, by pentafluorophenyl ester moieties to make the reaction with ammonia more efficient. To convert into such active ester moieties, the compound (4) maybe hydrolyzed, followed by the reaction with a substituted phenol such as pentafluorophenol.

This esterification reaction can be conducted preferably in the presence of a condensing agent such as N, N'-dichlorohexylcarbodiimide.

Step (b) In the step (b), ammonia is reacted with the compound (4) to afford the compound (5).

The reaction is conducted by reacting aqueous ammonia with the compound (4) in a solvent such as tetrahydrofuran.

As a reaction temperature, 0 to 80°C or so is sufficient.

Step (c) In the step (c), the compound (5) is reduced to yield

the compound (6).

This reducing reaction can be conducted preferably by <BR> <BR> <BR> <BR> using a boron hydride compound such as a boran-dimethyl sulfide complex. It is preferred to conduct the reaction in a solvent such as tetrahydrofuran at 0°C to the reflux temperature of the solvent.

Step (d) In the step (d), the compound (4) is reacted with the compound (6) to obtain the cyclic etheramine (la).

The reaction can be conducted preferably in an inert solvent, such as methylene chloride, in the presence of a base, such as triethylamine or pyridine, at 0°C to the reflux temperature of the solvent.

Step (e) In the step (e), the compound (la) is reduced to afford the compound (1b).

This reducing reaction can be conducted under similar conditions as in the above-described step (c).

Step (f) In the step (f), alkoxycarbonylalkylgroups (-R4-CooR5) are introduced onto the amino groups of the compound (1b), respectively, to obtain the compound (lc).

Illustrativeof arawmaterial for introducing the groups (-R4-COOR5) are a, ß-unsaturated fatty acid esters such as acrylic acid esters and p-halogenofatty acid esters such as

ß-bromopropionic acid esters.

When a ß-halogenofatty acid ester is used, the introduction can be effected as in the step (a). When an a, ß-unsaturated fatty acid ester is used, on the other hand, the introduction can be conducted preferably in the presence of a metal catalyst, such as cupric nitrate or cupric acetate, in a solvent, such as methanol, at a temperature of from 0 to 150°C.

Step (g) In the step (g), the compound (1c) is reduced to yield the compound (1d).

This reducing reaction can be conducted preferably by using a reducing agent such as lithium borohydride. It is preferred to conduct the reaction in a solvent, such as methanol or tetrahydrofuran, at room temperature to the reflux temperature of the solvent.

Step (h) In the step (h), the compound (ld) is halogenated to provide the compound (le).

As a halogenating agent, thionyl chloride or the like is used. The reaction can be conducted preferably in a solvent, such as methylene chloride, at 0 to 100°C.

Step (i) In the step (i), desired ones of various aryl groups and heterocyclic groups are introduced into the compound (le)

to obtain the compound (1f).

The reaction can be conducted preferably in the presence of a base such as dimethylaminopyridine.

As a method for isolating the target compound from the reaction mixture, it is preferred to use two or more of washing, recrystallization, various chromatographic techniques and the like in combination as needed. Further, the conversion into a salt can be conducted by a process known per se in the art.

The compound (1) of the present invention obtained as described above has effect to repair an abnormality in the <BR> <BR> <BR> <BR> <BR> expression of c-fos in cells of neuroblastoma origin stimulated by NGF, and is useful as a remedy for malignant tumors <BR> <BR> <BR> <BR> <BR> represented neuroblastomas in mammals including human beings.

The medicament according to the present invention can be obtained by adding to the compound (1) of the present <BR> <BR> <BR> <BR> <BR> invention one or more of pharmaceutically acceptable carriers such as excipients, binders, lubricants, disintegrators, coating agents, emulsifiers, suspending agents, solvents, stabilizers, absorption aids and ointment bases as needed and formulating the thus-obtained mixture into a preparation form for oral administration, injection administration or rectal administration or for external use by a method known per se in the art.

Preferred preparations for oral administration can

include granules, tablets, sugar-coated tablets, capsules, pills, liquids, emulsions and suspensions; preferred preparations for administration by injections can include intravenous injections, intramuscular injections, subcutaneous injections and instillations; and preferred preparations for rectal administration can include soft capsular suppositories.

The medicament according to the present invention can be administered to mammals, including human beings, as such preparations as described above.

The medicament according to the present invention can be administered preferably in a daily dose of from about 1 to 500 mg/kg one to four times a day.

Examples The present invention will next be described in further detail based on Examples. It should however be borne in mind that the present invention is by no means limited to or by these Examples. <BR> <BR> <BR> <BR> <BR> <P>Referential Example 1 Synthesis of CompoundA (in the formula<BR> <BR> <BR> <BR> <BR> <BR> (4), R3 = CH3) A mixture of 4, 4'-dihydroxydiphenylmethane (1.0 g, 5 mmol), methyl bromoacetate (1.53 g, 10 mmol) and potassium carbonate (1. 38 g, 10 mmol) in N, N-dimethylformamide (20 mL) was stirred at room temperature for 24 hours. The reaction

mixture was filtered, and the filtrate was extracted with ethyl acetate (50 mL x 3). The extract was washed with brine, and then dried over anhydrous magnesium sulfate. The solvent was <BR> <BR> <BR> <BR> <BR> evaporated under reduced pressure, and the residue was purified by column chromatography on silica gel (ethyl acetate: chloroform = 1: 9) to afford colorless needles (1.6 g, 93%). mp 64-65°C.

1H-NMR (CDCl3) b : 3.80 (s, 6H), 3.85 (s, 2H), 4.60 (s, 4H), 6.82 (d, 4H, J=8. 8Hz), 7.08 (d, 4H, J=8. 8Hz).

MS (EI) (m/z): 344 [M] +.

HRMS (EI) Calcd for Cl9H2006 : 344.1259.

Found 344.1256.

Referential Example 2 Synthesis of Compound B (in the formula <BR> <BR> <BR> <BR> <BR> (4), R3 = H)<BR> <BR> <BR> <BR> <BR> <BR> <BR> <BR> Amixture of CompoundA (1. 0g, 5 mmol) and 5 N KOH-methanol solution (4 mL) in methanol (40 mL) was refluxed for 2 hours.

The solvent was evaporated under reduced pressure, the residue was dissolved in water (100 mL), and the resultant solution was extracted with ethyl acetate (100 mL). The aqueous layer was acidified with 10% hydrochloric acid, followed by the extraction with ethyl acetate (300 mL). The extract was washed with brine and then dried over anhydrous magnesium sulfate.

The solvent was evaporated under reduced pressure to afford colorless powder (1.09 g, 100%).

mp 199-200°C.

H-NMR (DMSO-d6) b : 3.79 (s, 2H), 4.59 (s, 4H), 6.80 (d, 4H, J=8.

8Hz), 7.10 (d, 4H, J=8.8Hz), 12.90 (s, 2H).

MS (EI) (m/z): 316 [M] +.

HRMS (EI) (m/z): Calcd for C17H160g : 316.0946.

Found 316.0944.

Referential Example 3 Synthesis of Compound C (in the formula (4), R3 = pentafluorophenyl) A mixture of Compound B (2.95 g, 9.3 mmol), pentafluorophenol (3.46 g, 18.8 mmol) and N, N'-dicyclohexylcarbodiimide (3.88 g, 18.8 mmol) in tetrahydrofuran (100 mL) was stirred for 24 hours at room temperature. After the reaction mixture was filtered, the filtrate was evaporated under reduced pressure. The residue was purified by column chromatography on silica gel (dichloromethane) to afford colorless crystals (5. 56 g, 92%). mp 135-136°C.

H-NMR (CDCl3) b : 3.90 (s, 2H), 4.97 (s, 4H), 6.89 (d, 4H, J=8.4H z), 7.13 (d, 4H, J=8.4Hz).

MS (FAB) (m/z) : 648 [M] +.

HRMS (EI) (m/z): Calcd for C29H14F1006 : 648.0630.

Found 648.0628.

Referential Example 4 Synthesis of Compound D (Formula (5)) Amixture of Compound C (4. 0 g, 6.17 mmol) and 25% aqueous ammonia (12 mL) in tetrahydrofuran (30 mL) was stirred at room temperature for 12 hours. A saturated aqueous solution of

sodium hydrogencarbonate (200 mL) was added to the reaction mixture. The precipitate was collected by filtration, washed with water, ethanol and diethyl ether, and dried under vacuum to afford colorless crystals (1.9 g, 98%). mp 233-234°C.

1H-NMR (DMSO-d6) 3.80 (s, 2H), 4.36 (s, 4H), 6. 85 (d, 4H, J=8.

8Hz), 7.11 (d, 4H, J=8. 8Hz), 7.32 (s, 2H), 7.43 (s, 2H).

MS (FAB) (m/z): 315 [M+1] +.

HRMS (FAB) (m/z): Calcd for C17HlgN204 : 315.1344.

Found 315.1346.

Referential Example 5 Synthesis of Compound E (Formula (6)) A mixture of Compound D (314 mg, 1 mmol) and boran-dimethyl sulfide complex (1.16 mL, 12 mmol) in tetrahydrofuran (12 mL) was refluxed for 24 hours. A 0.7 M HCl-methanol solution (6 mL) was added, and the mixture was refluxed for 30 minutes. After the solvent was evaporated under reduced pressure, the residue was basified with 25% aqueous ammonia and then extracted with dichloromethane (300 mL). The extract was washed with brine and then dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure to obtain a pale yellow oil. The oil was purified by column chromatography on silica gel (chloroform: methanol: 25% aqueous ammonia = 100 : 40: 4) to afford a colorless amorphous solid (243 mg, 85%).

H-NMR (CD30D) 6 : 2.99 (t, 4H, J=5. 6Hz), 3.82 (s, 2H), 3.98 (t, 4 H, J=5.6Hz), 6.84 (d, 4H, J=8.4Hz), 7.07 (d, 4 H, J=8.4Hz).

MS (FAB) (m/z) : 287 [M+1] +.

HRMS (FAB) (m/z): Calcd for C17H23N202 : 287.1759.

Found 287. 1757.

Example 1 Synthesis of Compound F (Formula (la)) A mixture of Compound B (973 mg, 1.5 mmol), Compound E (430 mg, 1.5 mmol) and triethylamine (2.1 mL, 15 mmol) in dichloromethane (300 mL) was refluxed for 24 hours. After the solvent was evaporated under reduced pressure, the residue was purified by column chromatography on silica gel (ethyl acetate: methanol=9 : 1) to afford colorless crystals (586 mg, 69%). mp 195-196°C.

1H-NMR (CDCl3) 6 : 3.53 (s, 2H), 3.70-3. 73 (m, 4H), 3.80 (s, 2H), 3.92 (t, 4H, J=5.2Hz), 6.63 (d, 4H, J=8. 8Hz), 6.71 (d, 4H, J=8.8Hz), 6.87 (d, 4H, J=8.8Hz), 6.94 (t, 2H, J=5.2Hz), 7.05 (d, 4H, J=8. 8Hz).

MS (FAB) (m/z): 567 [M+1] +.

HRMS (FAB) (m/z): Calcd for C34H35N206 : 567. 2495.

Found 567.2496.

Example 2 Synthesis of Compound G (Formula (lb)) A mixture of Compound F (610 mg, 1.07 mmol) and boran-dimethyl sulfide complex (1.3 mL, 1.34 mmol) in THF (13 mL) was refluxed for 24 hours. A 0.7 M HCl-methanol solution

(6.5 mL) was added, and the mixture was refluxed for 30 minutes.

After the solvent was evaporated under reduced pressure, the residue was basified with 25% aqueous ammonia and then extracted with dichloromethane (100 mL). The extract was washed with brine and then dried over anhydrous sodium sulfate.

The solvent was evaporated under reduced pressure, and the residue was purified by column chromatography on silica gel (chloroform: methanol : 25% aqueous ammonia = 100: 10: 1) to afford colorless powder (480 mg, 83%). mp 135-136°C.

1H-NMR (CDCl3) 3. 0 0 (t, 8H, J=4.8Hz), 3. 80 (s, 4H), 4. 06 (t, 8 H, J=4.8Hz), 6.75 (d, 8H, J=8.4Hz), 7.00 (d, 8 H, J=8.4Hz).

MS (FAB) (m/z) : 539 [M+1] +.

HRMS (FAB) (m/z): Calcd for C34H3gN204 : 539.2909.

Found 539.2908.

Example 3 Synthesis of Compound H (in the formula (1c), R4 = (CH2)2, R5 = CH3) A mixture of Compound G (269 mg, 0.5 mmol), methyl acrylate (258 mg, 3 mmol) and cupric acetate monohydrate (10 mg, 0. 05 mmol) in methanol (5 mL) was stirred at 100°C for 24 hours. The reaction mixture was filtered through celite, and the filtrate was evaporated under reduced pressure. The residue was purified by column chromatography on silica gel (chloroform: methanol: 25% aqueous ammonia = 100: 10: 1) to

afford a colorless amorphous solid (305 mg, 86%).

H-NMR (CDCl3) # : 2.53 (t, 4H, J=7. 2Hz), 2. 95 (t, 8H, J=6. OHz), 2.99 (t, 4H, J=7.2Hz), 3.65 (s, 6H), 3.81 (s, 4 H), 3.94 (t, 8H, J=6. OHz), 6.72 (d, 8H, J=8.4H z), 7. 00 (d, 8H, J=8. 4Hz).

MS (FAB) (m/z) : 711 [M+1] +.

HRMS (FAB) (m/z): Calcd for C42H51N208 : 711.3645.

Found 711.3647.

Example 4 Synthesis of Compound I (in the formula (1d), R4 = (CH2) 2) A mixture of Compound H (300 mg, 0.42 mmol), lithium borohydride (84 mg, 3. 8 mmol) and methanol (0.24 mL) in tetrahydrofuran (12 mL) was refluxed for 24 hours. Water and 1 N hydrochloric acid were added to quench the reaction, then the reaction mixture was basified with 25% aqueous ammonia.

The resultant precipitate was collected by filtration, washed with water, and then dried under reduced pressure to afford colorless powder (260 mg, 95%). mp 182-183°C.

H-NMR (Pyridine-d5) 6 : 1. 91 (quint, 4H, J=6. 3Hz), 2.87 (t, 4H, J=6.3Hz), 2. 98 (t, 8H, J=5. 6Hz), 3.84 (s, 4H), 3.98 (t, 4H, J=6.3Hz), 4.07 (t, 8H, J=5.6Hz), 6.92 (d, 8H, J=8.4H z), 7.11 (d, 8H, J=8. 4Hz).

MS (FAB) (m/z): 655 [M+1] +.

HRMS (FAB) (m/z) : Calcd for C40H51N206 : 655.3746.

Found 655.3747.

Example 5 Synthesis of Compound J (in the formula (le), R4 = (CH2) 2, x2 = Cl) A mixture of Compound I (196 mg, 0.3 mmol) and thionyl chloride (2 mL) in dichloromethane (10 mL) was stirred at room temperature for 24 hours and was then evaporated under reduced pressure. The residue was diluted with water, and basified with saturated aqueous solution of sodium hydrogencarbonate.

The mixture was extracted with dichloromethane (25 mL), washed with brine and dried over anhydrous sodium sulfate. After removal of the solvent under reduced pressure, the residue was purified by column chromatography on silica gel (chloroform: methanol = 10: 1) to afford colorless powder (207 mg, 100%). mp 103-104°C.

1H-NMR (CDCl3) 6 : 1.95 (quint, 4H, J=6. OHz), 2. 78 (t, 4H, J=6. OH z), 2. 92 (t, 8H, J=6. 0Hz), 3.67 (t, 4H, J=6. OH z), 3.81 (s, 4H), 3.94 (t, 8H, J=6. 0Hz), 6. 72 (d, 8H, J=8.4Hz), 7.00 (d, 8H, J=8.4Hz).

MS (FAB) (m/z): 690 [M+11+, 692 [M+1] +37Cl, 694 [M+1] +37Cl2.

HRMS (FAB) (m/z): Calcd for C40H48Cl2N2O4 : 690.2990.

Found 690.2988.

Example 6 Synthesis of Compound K (in the formula (1f), R4 = (CH2) 2, R6 = 4-dimethylaminopyridin-1-yl) A mixture of Compound J (48 mg, 0.07 mmol) and

4-dimethylaminopyridine (17 mg, 0. 14 mmol) in methanol (2 mL) was refluxed for 18 hours. The reaction mixture was concentrated under reduced pressure. The resulting powder was washed with ethyl acetate, collected by filtration, and dried under reduced pressure to afford a colorless amorphous powder (65 mg, 100%).

1H-NMR (D20) 6 : 1.74 (brs, 4H), 2.13 (brs, 4H), 2.41 (brs, 20H), 3.26 (brs, 4H), 3.49 (brs, 8H), 3. 81 (brs, 4H), 5.85 (d, 4H, J=6.8Hz), 6.34 (d, 8H, J=8. OHz), 6.

66 (d, 8H, J=8. 0Hz), 7.48 (d, 4H, J=6.8Hz).

MS (FAB) (m/z) : 899 [M-Cl] +, 432 [M-Cl2] 2+.

HRMS (FAB) (m/z): Calcd for C54H6ôClN604 : 899. 4990.

Found 899.5074.

Compound K

Example 7 (1) As the cultured strains of neuroblastoma cells, two

strains (IMR-32, NB-39) were used. The compound of the present invention (the compound K) (10-5 M/L to 10-8 M/L) was caused to act on each strain, and 60 minutes later, NGF (100 ng/mL) was stimulated to act. Upon elapsed time of 60 minutes which were considered to allow the expression of c-fos to reach its peak, total RNA was extracted, cDNA was produced, and RT-PCT was performed. As a result, it was confirmed that for the IMR-32 strain known to have an abnormality in the expression of c-fos after NGF stimulation, addition of the cyclophane derivative according to the present invention (the compound K) achieved maximum expression of c-fos at a concentration as low as 10-8 M/L.

(2) An investigation was conducted about the persistence of reparative effect for the c-fos expression according to the present invention. Described specifically, the compound K (10-8 M/L) was caused to act on the IMR-32 strain, and 30 minutes later, NGF (100 ng/mL) was stimulated. The expression level of c-foswas determined with time by RT-PCR. As a result, it was found that the expression level of c-fos increased from 5.5 ng/mL shortly after NGF stimulation, to 6.7 ng/mL upon elapsed time of 3 hours after NGF stimulation, and to 8.1 ng/mL upon elapsed time of 7 hours after NGF stimulation and remained at 4.9 ng/mL and 5.5 ng/mL even 24 hours and 48 hours later, respectively. From the foregoing results, a peak was found to have reached 7 hours after NGF stimulation.

(3) An investigation was next conducted about c-fos expression reparative effect by advance administration of a compound according to the present invention. Described specifically, the compound K (10-8 M/L) was caused to act on the IMR-32 strain for 7 hours, NGF (100 ng/mL) was then caused to act, and the expression level of c-fos was determined with time by RT-PCR. As a result, it was found that the expression level of c-fos, which was 4.1 ng/mL shortly after NGF stimulation, apparently increased to 7.6 ng/mL one hour after NGF stimulation, remained at 5.1 to 5.7 ng/mL during 2 to 6 hours after the NGF stimulation, and clearly increased to 7.3 ng/mL 15 hours later, to 6.0 ng/ML 24 hours later, and to 12.1 ng/mL 48 hours later. Persistence of the effect of the compound K was hence confirmed. Similar effect was also exhibited on the NB-39 strain which is known to have an abnormality in the expression of c-fos.

Industrial Applicability The compounds (1) according to the present invention have reparative effect for an abnormality in the expression of c-fos, said abnormality being developed in neuroblastomas or the like, and are useful as remedies for various malignant tumors.