JP3341303B2 - Methylene diphosphonic acid derivative, its production method and its pharmaceutical use - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an in vivo
Suppresses the action of interleukin-1 that has an inflammation-inducing reaction, activation of various blood cells, bone destruction, etc., and at the same time suppresses the active oxygen that causes cell damage and fat degeneration, and also suppresses the effects of osteoporosis and rheumatoid arthritis disease. The present invention relates to a novel methylene diphosphonic acid derivative having an effect of suppressing bone destruction.

[0002]

2. Description of the Related Art Many diphosphonic acid compounds, which have been mainly developed as drugs for bone metabolism disorders, have an effect of suppressing bone destruction and are expected to suppress bone destruction at the onset of arthritis such as rheumatoid arthritis. Have been. JP-A-59-423
No. 95, JP-A-2-22285, JP-A-3-77894 and JP-A-60-174792 describe compounds having a diphosphonic acid structure.
These diphosphonic acid compounds are mainly used for inhibiting bone resorption and are effective as drugs for bone metabolic diseases, but are not yet sufficient for the treatment of rheumatoid arthritis. In order for diphosphonic acid compounds to be used for the treatment of rheumatoid arthritis,
At the same time as inhibiting bone resorption, it suppresses interleukin-1 (hereinafter abbreviated as IL-1), which is a mediator involved in inflammation, and suppresses cell damage caused by activated neutrophils and macrophages. There is a need for a new drug that has a combination.

[0003] IL-1 is known as a mediator involved in fever and inflammation, and its inhibitor has been expected as an anti-inflammatory drug. However, like many other cytokines, it is thought that IL-1 mainly acts locally, and many substances that suppress IL-1 in vitro have been reported. An anti-inflammatory drug having a sufficient effect of suppressing -1 and improving the disease state has not been developed. Infiltration of activated neutrophils and macrophages at the site of inflammation is observed during inflammation, and the active oxygen produced by these blood cells has the effect of digesting foreign substances, but normal when inflammation becomes chronic. It is known that even an organization can be impeded. Thus, compounds having an IL-1 inhibitory action and an antioxidant action are, of course, anti-inflammatory drugs,
It is also considered to be useful for autoimmune diseases such as rheumatoid arthritis and brain and liver organ disorders that occur during ischemia.

[0004]

DISCLOSURE OF THE INVENTION The present inventors have found that diphosphonic acid derivatives not only act as drugs for bone metabolism disorders,
By giving IL-1 inhibitory action and antioxidant action,
We have been studying diphosphonic acid compounds that exhibit excellent anti-inflammatory effects. In the course of this research, it was found that the addition of a naphthalene skeleton to the diphosphonic acid structure has an IL-1 inhibitory effect and an antioxidant effect that are not present in existing drugs. The present invention relates to interleukin-1
It is an object of the present invention to provide a useful novel compound having an action of suppressing lipase, an antioxidant action and an action of inhibiting bone destruction.

[0005]

The above objects are achieved by the present invention described below. That is, the present invention provides a compound represented by the general formula (I):
Methylene diphosphonic acid derivative shown in

Embedded image [In the formula, X represents a substituent on a naphthyl group, a halogen, a nitro group, a nitrile group, a trifluoromethyl group,

Embedded image Group (provided that Z ¹ and Z ² independently represent a hydrogen atom or an alkyl group, or Z ¹ and Z ² may form a carbon ring or a carbon ring containing a heteroatom) ),

Embedded image Groups (wherein Z ¹ and Z ² are the same as above and Z ³ represents oxygen or sulfur), thiol group, alkylthio group, arylthio group, acylamino group, acylthio group, acyl group, alkenyl group, aryl group , Cycloalkyl group, COOH group or COO alkyl group, m represents an integer of 0 to 3, Y represents a substituent on a naphthyl group, halogen, nitro group, nitrile group, alkyl group, alkoxy group , Trifluoromethyl group,

Embedded image A group wherein Z ¹ and Z ² are the same as above;

Embedded image Groups (however, Z ¹ , Z ² and Z ³ are the same as described above), thiol group, hydroxyl group, alkylthio group, arylthio group, acyloxy group, acylamino group, acylthio group, acyl group, alkenyl group, aryl group, cyclo group An alkyl group, a COOH group, or a COO alkyl group, n represents an integer of 0 to 4, m X and n Y
May be the same or different, A is-(CH
2) a-S- (a is Ri integer der of 0, A sulfur atom
Is directly bonded to methanebisphosphonic acid ), B means hydrogen, R ¹ , R ² , R ³ and R ⁴ are hydrogen, carbon atom 1
-7 linear or branched alkyl groups or pharmacologically acceptable cations, which may be the same or different. And a pharmaceutical use of the derivative as an active ingredient, such as an anti-inflammatory drug, an anti-rheumatic drug, a drug for bone metabolism, an interleukin-1 inhibitor, an antioxidant, and a bone destruction inhibitor. Things.

An unsubstituted naphthyl group represents a 1-naphthyl group or a 2-naphthyl group, and a naphthyl group having one or more substituents has 1-naphthyl having a substituent at the 2- to 8-position. Represents a naphthyl group or a 2-naphthyl group having a substituent at the 1- or 3-position to the 8-position. When the naphthyl group has a substituent, the preferred position of the substituent is, when the naphthyl group is a 1-naphthyl group, the position substituted with X is the 2-position and / or 4-position, and the position substituted with Y is In the 5-position and / or 6-position and / or 8-position, when the naphthyl group is a 2-naphthyl group, the position substituted with X is 1
Position and / or position 4;
Position and / or 6 position and / or 8 position. Halogen atoms used as the substituents X and Y are fluorine, chlorine, bromine and iodine. The alkyl group (the alkyl groups shown below have the same meaning) is a linear or branched one having 1 to 7 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl,
Pentyl, hexyl, cyclohexylmethyl and the like can be mentioned. The alkoxy group has 1 to 7 carbon atoms and includes, for example, methoxy, ethoxy, propoxy, isopropoxy, butoxy and the like.

[0007]

Embedded image Examples of the group (the alkyl of Z ¹ and Z ² are the same as described above) include amino, methylamino, ethylamino, propylamino, butylamino, dimethylamino, diethylamino, pyrrolidino, piperidino, morpholino, thiomorpholino and the like.

[0008]

Embedded image The groups (Z ¹ , Z ² and Z ³ are as defined above) are carbamoyl, thiocarbamoyl, N-methylaminocarbonyl,
N, N-dimethylaminocarbonyl, piperidinocarbonyl, pyrrolidinocarbonyl, morpholinocarbonyl and the like can be mentioned. Examples of the alkylthio group (alkyl is the same as the above-mentioned alkyl group) include methylthio, ethylthio, propylthio, isopropylthio, cyclopentylthio, cyclohexylthio and the like. An arylthio group is
It preferably has 6 to 15 carbon atoms, such as phenylthio and substituted phenylthio. The acyl (group) of acyloxy, acylamino, acylthio, and acyl groups has 2 carbon atoms.
And 7 linear or branched chains, for example, acetyl, propanoyl, butanoyl and the like. The alkenyl group has a straight or branched chain having 2 to 7 carbon atoms and includes vinyl, allyl, 2-propenyl, isopropenyl, butenyl, pentenyl and the like. The aryl group preferably has 6 to 15 carbon atoms, phenyl, substituted phenyl,
And naphthyl. A cycloalkyl group has 3 to 3 carbon atoms.
8 and includes cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and the like. Examples of the COO alkyl group (alkyl is the same as the above-mentioned alkyl group) include methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl and the like.

A is-(CH ₂ ) _a- (D) _b- (C
H ₂ ) _c- (D is sulfur, oxygen, NH or alkyl-substituted N, a and c are integers from 0 to 10, and b is 0 or 1 (however, when b = 0, a = c = 0)), and more preferably a, b and c are independently 0 or 1.

Further, when B is other than hydrogen or an alkyl group, those having C = 0 are not preferable because they are chemically unstable. However, even in this case, a = b = c = 0 is stable and preferable.

In a particularly preferred embodiment, A is S,
O, NH, CH ₂ S, CH ₂ O, CH ₂ NH, SC
H ₂ , OCH ₂ , NHCH ₂ and the like. Further, the naphthyl group is not mediated by A (that is, the case where a = b = c = 0),
Also included are compounds directly connected to the carbon of methylene diphosphonic acid.

When B is a monoalkylamino group, a dialkylamino group or an alkoxy group, the alkyl is the same as the above-mentioned alkyl group, and the acyl of the acylamino group and the acyloxy group is the same as the above-mentioned acyl.

Representative alkyl groups for R ¹ , R ² , R ³ and R ⁴ include methyl, ethyl, propyl,
Isopropyl, butyl, isobutyl, t-butyl, pentyl and the like.

When R ¹ , R ² , R ³ and R ⁴ are hydrogen, the phosphonic acid moiety of formula (I) can form salts with inorganic or organic bases. In this case, the pharmacologically acceptable cation includes a metal cation and ammonium NR ₄ (where R is the same as the above-described alkyl group and hydrogen of R ^{1 to} R ⁴ ), and particularly preferred metal cations. Are cations of alkaline earth metals, for example, magnesium, calcium and the like. However, other metals, such as cations such as aluminum, zinc, iron, etc., are also included in the present invention. Ammonium includes ammonium, primary amine, secondary amine, tertiary amine ammonium and quaternary ammonium. These include ammonia, methylamine, dimethylamine, trimethylamine, ethylamine, diethylamine, triethylamine, propylamine, dipropylamine, isopropylamine, diisopropylamine, butylamine, dibutylamine,
Examples include ammonium such as isobutylamine, t-butylamine, monoethanolamine, diethanol, and triethanolamine, and tetramethylammonium and tetraethylammonium. Of these, cations of sodium, potassium, ammonia and alkylamine are preferred. In R ^{1 to} R ⁴ , the cations may be the same or different, and a mixture of cations and hydrogen, for example, monocationic salts, dicationic salts, and tricationic salts are also included in the present invention. Preferably, methylene diphosphonic acid derivative represented by the general formula (I), the ones that all R ¹ to R ⁴ is hydrogen, or two are hydrogen of R ¹ to R ^4, the rest two are sodium Thing or 2
One is hydrogen and the other two are ammonium.

The methylene diphosphonic acid derivative of the present invention comprises
It can be produced by a method similar to a method known in the art. For example, one of the methylene diphosphonic acid derivatives of formula (I) of the present invention (when B is H)
Can be produced by a method represented by the following reaction formula.

[0016]

Embedded image The starting material used is a lower alkyl ester of methylene diphosphonic acid) (II) (where lower alkyl is a straight-chain or branched-chain alkyl having 1 to 7 carbon atoms), sodium hydride, alkyl By reacting with a base such as lithium, the corresponding metalated methylene diphosphonic ester (V) is obtained, to which various naphthyl-A group introducing agents (where naphthyl is

Embedded image (X, Y, m, n are the same as above, and A is the same as defined above) to give compound (VI). Naphthyl
[Naphthyl- (CH ₂ ) _a -S] ₂
(A is the same as described above) is used.

The reaction temperature and reaction time depend on the reagents used. For example, the reaction temperature is between -78 C and the boiling point of the solvent or solvent mixture, and the reaction time ranges from 10 minutes to several days.

A methylene diphosphonic acid derivative in which R ^{1 to} R ⁴ is hydrogen is obtained by hydrolysis or the like from a methylene diphosphonic acid (phosphonic ester) in which R ^{1 to} R ⁴ is an alkyl group. For example, phosphonates can be reacted with an acid such as hydrochloric acid, or trimethylsilyl bromide,
It is then hydrolyzed by treatment with water or alcohol. The methylene diphosphonic acid thus obtained can be converted into one of its salts in a known manner.

Compounds in which 1 to 3 of R ^{1 to} R ⁴ are alkyl groups by partial hydrolysis of methylene diphosphonic acid ester or partial esterification of methylene diphosphonic acid (methylene diphosphonate) Partial esters of acids) are also included in the present invention.

Most of the methylene diphosphonic acid derivatives of the present invention have a P ＝ O bond as a keto type, but partially exist as an enol type depending on the chemical properties of the compound itself, an external environment such as a solvent and temperature. In some cases, these are also included in the compounds of the present invention.

When all the reactions contain reactive substituents and reactive functional groups other than the target reaction, these substituents and functional groups are blocked in advance by a reagent which can be easily removed. Must be kept.

The diseases targeted by the compounds of the present invention include inflammatory diseases, pain diseases, skin diseases, respiratory diseases, liver diseases,
Infectious disease, autoimmune disease, ischemic organ damage or bone metabolism disease. For example, (chronic) rheumatoid arthritis, rheumatoid polyarthritis, osteoarthritis, scapular periarthritis, cervico-brachial syndrome, intervertebral disc disorders, low back pain, tendon / tenosynovitis, osteoarthritis, fifty shoulder, fibrospermitis, myalgia , Neuralgia, gout, inflammation and swelling after surgery and trauma (anti-inflammatory, anti-rheumatic, anti-arthritic, analgesic and antipyretic) or psoriasis, asthma, pulmonary sarcoidosis, viral hepatitis, human immunity Deficiency virus infection, protozoan infection, ischemic heart disease, ischemic encephalopathy, ischemic liver injury, arteriosclerosis and osteoporosis, Paget's disease, Behitilev's disease, hypercalcemia, ectopic ossification, etc. Metabolic bone disease drug).

When the novel methylene and methanediphosphonic acid derivatives of the present invention are used in the above-mentioned applications of the present invention, a pharmaceutical composition as it is or mixed with a pharmaceutically acceptable carrier, excipient or the like known per se. It is used as an object. Administration may be any of oral administration such as tablets, capsules, powders, granules and pills, and parenteral administration such as injections, syrups, ointments and suppositories. The dose is
The dose is about 0.1 mg to 5 g, preferably about 1 mg to 2 g, depending on the administration subject, administration route, symptoms and the like, and is orally or parenterally administered once or several times a day.

[0024]

The present invention will now be described more specifically with reference to examples.

Example 1 Tetraethyl 2-naphthylthiomethylenediphosphonate
(1)

Embedded image Under an argon atmosphere, a solution of 10.09 g (35 mmol) of tetraethyl methylenediphosphonate in dry tetrahydrofuran (100 ml) was cooled to -78 ° C, and n-
Butyllithium hexane solution [1.59mol / l] 2
2.01 ml (35 mmol) was added and stirred for 30 minutes. Next, a solution of 11.15 g (35 mmol) of 2,2′-dinaphthyl disulfide in 75 ml of dry tetrahydrofuran was added to the mixture, and the temperature was raised to room temperature.
Stirred for 16 hours. Pour the resulting mixture into ice water,
After neutralization with 6N hydrochloric acid, ethyl acetate (3 × 150 m
Extracted in l). The organic layer was washed with water and saturated saline, dried over anhydrous magnesium sulfate, the solvent was distilled off under reduced pressure, and the obtained residue was purified by column chromatography (developing solvent: ethanol: ethyl acetate = 5: 95). Then, 8.82 g of the title compound was obtained as a colorless oil. Yield 57%.

¹ H-NMR (CDCl ₃ ) [ppm]:
1.33 (t, J = 7Hz, 12H), 3.55 (t, J = 21Hz, 1H), 4.00 ~ 4.55 (m, 8
H), 7.46 to 7.52 (m, 3H), 7.76 to 7.83 (m, 3H), 8.07 (S, 1H) IR (KBr) [cm ^-1 ]: 2984, 1626, 1589, 1502, 139
2,1257,1029,975 MASS (FAB) m / z: 447 (M + H) ⁺ EA (as C ₁₉ H ₂₈ O ₆ P ₂ S) Calculated value (%): C 51.12 H 6.33 Measured value ( %): C 51.10 H 6.29

Example 2 2-Naphthylthiomethylene diphosphonic acid (2)

Embedded image Under an argon atmosphere, 8.04 g of tetraethyl 2-naphthylthiomethylenediphosphonate obtained in Example 1 (18
mmol) in dry methylene chloride (100 ml) solution,
At room temperature, 27.56 g of brominated trimethylsilane (180 mm
ol) was added dropwise, and the mixture was stirred at room temperature for 72 hours. After distilling off the solvent and excess trimethylsilane bromide under reduced pressure, the obtained residue was dissolved in a mixed solution of water: methanol = 5: 95, heated and refluxed for 30 minutes, and the solvent was distilled off again under reduced pressure. . The obtained residue was crystallized using acetone-methylene chloride as a solvent, and the obtained crystals were recrystallized again from the same solvent system to obtain the title compound as white crystals. Yield 86
%.

M. p. : 218.5-219.5 ° C (d
ec) ¹ H-NMR (CD ₃ OD) [ppm]: 3.51 (t, J = 21H)
z, 1H), 7.42-7.51 (m, 2H), 7.65-7.70 (m, 1H), 7.74-7.8
7 (m, 3H), 8.10 to 8.12 (m, 1H) IR (KBr) [cm ^-1 ]: 2926,1657,1638,1620,115
1,973,812 MASS (FAB) m / z : 335 (M + H) + EA (C 11 H 12 O 6 P 2 S) Calculated (%): C 39.53 H 3.63 measurements (%): C 39. 62H 3.70

Example 3 Tetraethyl 6-methoxy-2-naphthylthiomethylenediphosphonate (3)

Embedded image (A) 6,6′-dimethoxy-2,2′-dinaphthyldi
Under a sulfide argon atmosphere, 6,6′-dihydroxy-2,2 ′
10.00 g of dinaphthyl disulfide (28.5 mm
ol) in dry dimethylformamide (250 ml) was cooled to -23 ° C and sodium hydride (60%
3.42 g of mineral oil dispersion (85.
(5 mmol) was added slowly, and the mixture was stirred until hydrogen evolution ceased. Next, 12.14 g of methyl iodide was added to the mixture.
(85.5 mmol) was added, and the temperature was raised to room temperature.
Stirred for hours. The resulting mixture was poured into ice water and extracted with ethyl acetate (3 × 150 ml). The organic layer was washed with water and saturated saline, dried over anhydrous magnesium sulfate, and then the solvent was distilled off under reduced pressure. The obtained crystals were recrystallized from ethyl acetate-petroleum ether to give 9.92 g of 6,6′-dimethoxy-2,2′-dinaphthyl sulfide as orange crystals. Yield 92%.

M. p. : 125 to 126 ° C ¹ H-NMR (CDCl ₃ ) [ppm]: 3.89 (s, 6H),
7.00 to 7.25 (m, 4H), 7.45 to 7.75 (m, 6H), 7.85 to 7.95 (m, 4H)
2H)

(B) 6-methoxy-2-naphthylthiome
According to a method similar to that of Example 1, 10.09 g (35 mmol) of tetraethyl methylenediphosphonate and 6,6′-
12. dimethoxy-2,2'-dinaphthyl disulfide
From 25 g (35 mmol), 11.34 g of the title compound
Was obtained as a pale yellow oil. Yield 68%.

¹ H-NMR (CDCl ₃ ) [ppm]:
1.33 (t, J = 7Hz, 6H), 1.35 (t, J = 7Hz, 6H), 3.48 (t, J = 22Hz, 1
H), 3.91 (s, 3H), 4.00 ~ 4.45 (m, 8H), 7.00 ~ 7.30 (m, 2H),
7.50 to 7.80 (m, 3H), 8.00 to 8.10 (m, 1H) IR (KBr) [cm ^-1 ]: 2984,2936,2910,1628,159
3,1390,1259,1214,1023,975 MASS (FAB) m / z : 477 (M + H) + EA (C 20 H 30 O 7 P 2 as S) Calculated (%): C 50.42 H 6 . 36 measured value (%): C 50.65 H 6.42

Example 4 6-Methoxy-2-naphthylthiomethylene diphosphonic acid
(Four)

Embedded image In the same manner as in Example 2, 6-
7.15 g (15 mmol) of tetraethyl methoxy-2-naphthylthiomethylene diphosphonate was treated with trimethylsilane bromide in dry methylene chloride and then hydrolyzed to obtain 4.21 g of the title compound as white crystals. Yield 77%.

M. p. : 234 to 235 ° C (dec) ¹ H-NMR (CD ₃ OD) [ppm]: 3.37 (t, J = 21H)
z, 1H), 3.90 (s, 3H), 7.12-7.23 (m, 2H), 7.65-7.75 (m, 3
H), 8.05-8.09 (m, 1H) IR (KBr) [cm ^-1 ]: 2920,1628,1466,1214,112
5,994,924,911 MASS (FAB) m / z : 365 (M + H) + EA (C 12 H 14 O 7 P 2 as S) Calculated (%): C 39.57 H 3.88 measurements (%): C 39. 55H 3.79

Example 5 Tetraethyl 6-hydroxy-2-naphthylthiomethylenediphosphonate (5)

Embedded image (A) 6,6′-di (t-butyldimethylsiloxy)-
2,6'-Dihydroxy-2,2 'under an argon atmosphere of 2,2'-dinaphthyl disulfide
10.00 g of dinaphthyl disulfide (28.5 mm
ol) and 9.70 g (140 mmol) of imidazole
1) dimethylformamide (150 ml) solution at 0 ° C
And then t-butyldimethylchlorosilane 1
A solution of 2.89 g (85.5 mmol) in dry dimethylformamide (50 ml) was added, and the mixture was heated to room temperature.
Stir for 3 hours. The resulting mixture was poured into ice water and extracted with ethyl acetate (3 × 150 ml). The organic layer was washed with water and saturated saline, dried over anhydrous magnesium sulfate, and then the solvent was distilled off under reduced pressure. The obtained residue was purified by column chromatography (developing solvent: ethyl acetate: n-hexane = 5: 95) to obtain the title disulfide compound as a pale yellow oil. Yield 99%.

¹ H-NMR (CDCl ₃ ) [ppm]:
0.23 (s, 6H), 1.00 (s, 9H), 6.95-7.20 (m, 2H), 7.45-7.70
(m, 3H), 7.85 to 7.95 (m, 1H)

(B) 6-hydroxy-2-naphthylthio
Tetraethyl methylenediphosphonate In the same manner as in Example 1, 10.09 g (35 mmol) of tetraethyl methylenediphosphonate and 6,6′-
The reaction with 13.25 g (35 mmol) of di (t-butyldimethylsiloxy) -2,2'-dinaphthyl disulfide was carried out, and the resulting residue was mixed without isolation and 6N hydrochloric acid: methanol = 1: 20. The mixture was dissolved in the solution, heated and stirred at 50 ° C. for 3 hours, and the solvent was distilled off again under reduced pressure. The obtained residue was purified by column chromatography (developing solvent: ethanol: ethyl acetate = 5: 95) to obtain 10.02 g of the title compound as a pale yellow oil which gradually crystallized. Yield 62%.

M. p. : 85.5-86.5 ° C ¹ H-NMR (CDCl ₃ ) [ppm]: 1.35 (t, J = 7H
z, 12H), 3.50 (t, J = 22Hz, 1H), 4.05-4.65 (m, 8H), 6.95-
7.20 (m, 2H), 7.25 to 7.70 (m, 3H), 7.90 to 8.05 (m, 1H), 8.9
5 (brs, 1H) IR (KBr) [cm ^-1 ]: 3148,2984,1626,1392,123
2,1212,1027 MASS (FAB) m / z: 463 (M + H) ⁺ EA (as C ₁₉ H ₂₈ O ₇ P ₂ S) Calculated value (%): C 49.35 H 6.12 Measured value (%) : C 49.39 H 6.11

Example 6 6-Hydroxy-2-naphthylthiomethylene diphosphonic acid (6)

Embedded image In the same manner as in Example 2, 6-
By treating 7.15 g (15 mmol) of tetraethyl hydroxy-2-naphthylthiomethylene diphosphonate with trimethylsilane bromide and then hydrolyzing,
4.21 g of the title compound were obtained as white crystals.

[0040] Yield 75%.

M. p. : 210 to 211 ° C ¹ H-NMR (D ₂ O) [ppm]: 3.51 (t, J = 20 Hz, 1
H), 7.02 ~ 7.13 (m, 2H), 7.48 ~ 7.55 (m, 1H), 7.55 ~ 7.62
(m, 1H), 7.63 to 7.69 (m, 1H), 7.90 to 7.97 (m, 1H) IR (KBr) [cm ^-1 ]: 3570, 3164, 1636, 1506, 113
5,1046,939,919 MASS (FAB) m / z: 351 (M + H) ⁺ EA (as C ₁₁ H ₁₂ O ₇ P ₂ S) Calculated value (%): C 37.73 H 3.46 Measured value (%) : C 37.80 H 3.55

Example 7 Tetraethyl 1-naphthylthiomethanediphosphonate (7)

Embedded image (A) 1,1′- Dinaphthyl disulfide Iodotrimethylsilane 10 was added to a solution of 22.67 g (100.0 mmol) of 1-naphthalenesulfonyl chloride in 250 ml of dry methylene chloride under an argon atmosphere.
0.0 g (500.0 mmol) was slowly added, and the mixture was stirred for 6 hours. The resulting mixture was poured into a saturated aqueous sodium hydrogen carbonate solution and extracted with methylene chloride (3 × 150 ml). The organic layer was washed with a saturated aqueous solution of sodium thiosulfate until the iodine color disappeared, further washed with water and a saturated saline solution, and then dried over anhydrous magnesium sulfate.
The solvent was distilled off under reduced pressure, and the resulting crystals were collected in ethyl acetate-n-
Recrystallization from hexane gave 12.43 g of the title compound as pale yellow crystals. Yield 78%.

M. p. : 85 to 86 ° C ¹ H-NMR (CDCl ₃ ) [ppm]: 7.15 to 8.00
(m, 12H), 8.20-8.45 (m, 2H)

( B) 1-naphthylthiomethanediphosphone
In a similar manner to that orthosilicate Example 1, methylene diphosphonic acid tetraethyl 10. . 9g (35mmol) and 1,1'-
11.15 g (35 mmol) of dinaphthyl disulfide
To give 9.38 g of the title compound as a pale yellow oil.
Yield 60%.

¹ H-NMR (CDCl ₃ ) [ppm]:
1.30 (t, J = 7Hz, 12H), 3.55 (t, J = 21Hz, 1H), 3.95-4.45 (m, 8
H), 7.30 to 7.75 (m, 3H), 7.75 to 8.10 (m, 3H), 8.55 to 8.75
(m, 1H) IR (KBr) [cm ^-1 ]: 3434,2984,2934,2910,150
6,1444,1255,1164,1098,1013,971 MASS (FAB) m / z: 447 (M + H) ⁺ EA (as C ₁₉ H ₂₈ O ₆ P ₂ S) Calculated value (%): C 51.12 H 6.33 measured value (%): C 51.33 H 6.19

Example 8 1-Naphthylthiomethanediphosphonic acid (8)

Embedded image In the same manner as in Example 2, 6.70 g (15 mmol) of tetraethyl 1-naphthylthiomethylenediphosphonate was used.
l) was treated with trimethylsilane bromide and then hydrolyzed to give 3.92 g of the title compound as white crystals. Yield 78%.

M. p. : 241-242 ° C (dec) ¹ H-NMR (CD ₃ OD) [ppm]: 3.45 (t, J = 21H)
(z, 1H), 7.30 ~ 7.70 (m, 3H), 7.75 ~ 8.10 (m, 3H), 8.60 ~ 8.
80 (m, 1H) IR (KBr) [cm ^-1 ]: 2910,2892,1506,1185,114
1,1006,932 MASS (FAB) m / z: 335 (M + H) ⁺ EA (as C ₁₁ H ₁₂ O ₆ P ₂ S) Calculated value (%): C 39.53 H 3.63 Measured value (%) : C 39.44 H 3.70

Example 9 Adjuvant Arthritis Test Injection of Mycobacterium tuberculosis adjuvant into rats results in polyarthritis resembling human rheumatoid arthritis. Using this adjuvant arthritis model, the effects of the compound of the present invention on anti-inflammatory / anti-rheumatic and bone metabolism improvement were examined by the following procedure.

0.1 mg of dried dead mycobacterium butyricum was suspended in 0.1 ml of liquid paraffin and injected into the footpad of the left hind leg of a 7-week-old Lewis female rat. The compounds obtained in the examples were dissolved in sterile distilled water and subcutaneously administered at a ratio of 20 mg / kg body weight for two consecutive weeks from day 8 to day 21 from the day of adjuvant injection. During this time, measure the foot volume of the left and right hind limbs,
The edema rate was calculated by the following equation.

[0050]

(Equation 1) Further, the edema inhibition rate was determined by the following equation, and is shown in Table 1.

(Equation 2)

On the 22nd day, the rats were sacrificed, and soft X-ray photographs of the left and right hind limbs were taken. Left and right hind limbs 5 based on soft X-ray
The degree of bone destruction at each of the four locations was evaluated with a five-point score, and the total number was defined as the bone destruction score. Further, the rate of inhibiting bone destruction was calculated by the following equation, and is shown in Table 1.

[0052]

(Equation 3)

The results obtained are shown by the Student's t-test and the Tukey's multiple comparison method, where the risk factor is P <0.001 and the significance is P ** 1 for the control group to which only sterile distilled water was administered. And ** for P <0.01 significant **
Marks are given, and those with significance at P <0.05 are marked with *. As is clear from Table 1, the compound according to the present invention suppressed foot edema and bone destruction due to primary and secondary inflammation of adjuvant arthritis.

[0054]

[Table 1]

Example 10 IL- from mouse macrophage strain cell J774-1
1 Action on production Macrophages, which are lymphocytes, phagocytose invading microorganisms and blood cell debris as a foreign body elimination mechanism,
It presents antigens to cells and releases active oxygen to digest foreign substances. At this time, macrophages release various cytokines. Among them, IL-1 is fever, inflammation,
It is known that it has effects such as cartilage / bone destruction, activation of leukocytes, damage to vascular endothelial cells, and various effects by inducing other cytokines.

Mouse macrophage cell line J774-1
Is a cell line selected from those exhibiting high IL-1 production, and is known to exhibit IL-1 production when stimulated with LPS. Using this cell line, the inhibitory effect of the compound of the present invention on IL-1 production was measured by the following procedure.

J774-1 cells were subjected to RPM containing 10% fetal calf serum and 50 μM of 2-mercaptoethanol.
The cells were cultured in I-1640 medium and adjusted to a cell number of 2 × 10 ⁵ cells / ml. 1 ml of the cell suspension was placed in a 24-well plate and cultured for 30 minutes. Thereafter, LPS was added to a final concentration of 1 μg / ml, and at the same time, the compound obtained in the example was dissolved in sterilized distilled water to obtain 100 μl.
M was added at a concentration of M. 2 at 37 ° C, 5% CO ₂ environment
After culturing for 4 hours, the supernatant was collected, centrifuged to remove cell debris and the like, and then sterilized through a 0.22 μm filter.

The activity of IL-1 was measured using C3H / He J
It was measured by the thymocyte proliferation activity of male mice. That is, C3
Thymus was collected from 4-6 week old H / He J male mice. The thymus was loosened in RPMI-1640 medium containing 10% fetal calf serum and 50 μM of 2-mercaptoethanol to prepare a cell solution at a concentration of 2 × 10 ⁷ cells / ml. Phytohemagglutinin was added to this cell suspension to a final concentration of 1%, and this was used as a T cell suspension.

The sample obtained above was subjected to two-fold serial dilution in a 96-well multiplate in a volume of 50 μl, and a T cell suspension was added thereto in an amount of 50 μl each. T
The cells were cultured for 72 hours, and the IL-1 activity was determined based on the cell growth rate. The cell growth was performed at 4- [4,5
-Dimethylthiazol-2-yl] -2,5-diphenyltetrazolium bromide, and the maximum of T cells with recombinant human IL-1 was determined using the absorbance at 570 nm of a dye that was reduced by viable cell mitochondria to form a color. The degree of proliferation was defined as 100%,
The dilution ratio of the sample when the growth of T-saibo was caused by 50% assuming that the growth without addition of -1 was 0% was calculated as the number of units of the sample.

At this time, the inhibition rate of the compound of the present invention on IL-1 production when LPS of J774-1 cells was stimulated with 1 μg / ml of LPS was calculated by the following formula. Table 2 shows the results.

(Equation 4)

[Table 2]

Example 11 When cartilage of a rabbit knee joint was isolated and cultured and stimulated with IL-1, proteoglycan, a glycoprotein which is a main component of chondrocytes, was released. Using this effect as an index, the IL-1 inhibitory effect of the compound of the present invention was measured as follows.

A New Zealand white male rabbit, 3 weeks old, weighing 250-300 g, was bled to death under diethyl ether anesthesia, and the knee joint was separated. Cut out the cartilage from the knee joint with a scalpel, 0.14M
Sodium chloride, 4 mM potassium chloride, 0.4 mM sodium dihydrogen phosphate, 12 mM sodium bicarbonate,
It was immersed in a CMF solution consisting of 0.2 mM potassium dihydrogen phosphate and 11 mM glucose. 0.1% of this cartilage
The plate was placed in EDTA and incubated at 37 ° C. for 20 minutes.
After removing the supernatant, 0.15% trypsin was added and 37
Incubated at 60 ° C for 60 minutes. After washing three times with a CMF solution, 0.15% collagenase was added thereto, and further at 37 ° C.,
Treated for 05 minutes. A piece of cartilage tissue was isolated into chondrocytes by pipetting, passed through a 120 μm nylon mesh, and centrifuged at 4 ° C., 500 g for 7 minutes to obtain chondrocytes.
The cells are washed three times and Dulbecco M containing 10% fetal bovine serum
The cells were suspended in an EM medium at 1.2 × 10 ⁵ cells / ml. 250 μl of the cells were placed in a 48-well plate and cultured for 5 days until confluence was reached. Thereafter, the culture solution was added to Dulbecco ME containing 0.3% fetal bovine serum.
After replacing with M and further culturing for 24 hours, ³⁵ S-labeled inorganic sulfuric acid was added at a concentration of 185 kBc and cultivated for 24 hours. After washing the cells three times with Dulbecco's MEM medium, the medium was washed with BGjb containing 0.1% bovine serum albumin.
The medium was exchanged, and 30 g of the recombinant human IL-1β was added.
It was added at a concentration of units / ml. At the same time, the compounds of the invention were dissolved in sterile distilled water and added at a final concentration of 100 μM. Twenty-four hours after the IL-1 stimulation, the culture supernatant and the cell layer were collected.

The cell layer was decomposed by adding 200 μg of pronase E and treating at 37 ° C. for 24 hours. 0.05 ml of 0.1 mg / ml chondroitin sulfate was added to the culture supernatant,
0.5 ml of 2 mM magnesium sulfate, 0.2 M Tris-HCl buffer solution of 5 mM calcium chloride (pH 7.8)
0.5 ml of a 1 mM cetylpyridinium chloride 20 mM sodium chloride solution was added sequentially, and the proteoglycans precipitated by treatment at 37 ° C. for 2 hours were collected on a glass filter, a liquid scintillator was added, and the mixture was counted with a liquid scintillation counter.

In the cell layer, 0.05 ml of 0.1 mg / ml chondroitin sulfate and 0.2 mM of 2 mM magnesium sulfate were added.
5 ml of 1% cetylpyridinium chloride 20 mM sodium chloride solution 0.5 ml was sequentially added, and the proteoglycan precipitated by treatment at 37 ° C. for 2 hours was collected on a glass filter, added with a liquid scintillator, and counted with a liquid scintillation counter.

The counts obtained in each case were expressed as a percentage divided by the count of the inorganic sulfuric acid initially added.
The results obtained were determined using the Student's t-test, and those with a significance level P <0.01 relative to the unstimulated control group
Marks indicate the risk ratio P <0.01 relative to the IL-1 stimulated control group.
** indicates significant ones. As shown in Table 3, the compounds of the present invention inhibit proteoglycan release from the cell layer upon IL-1 stimulation, and are useful as IL-1 inhibitors.

[0066]

[Table 3]

Example 12 It is known that neutrophils phagocytose foreign substances to eliminate them and produce active oxygen and digestive enzymes as a biological defense reaction. However, during chronic inflammation or the like, it is conceivable that active oxygen and digestive enzymes produced by neutrophils damage normal tissues and further enhance inflammation. Therefore, the effect of the compound of the present invention on the release of active oxygen from human neutrophils was measured as follows.

Using 3.8% sodium citrate as an anticoagulant, 50 ml of blood was collected from a human vein. Equal amounts of this blood and 2% dextran saline solution are mixed,
After shaking several times, it was left still at 37 ° C. for 30 minutes. The upper layer was separated and overlaid on an equal volume of Ficoll-Paque solution. 20 ° C, 1
The precipitate obtained after centrifugation at 400 rpm for 30 minutes is taken, the cells are resuspended in Hanks buffer, and
The cells were washed by spinning and centrifuging for 5 minutes. The mixed erythrocytes are removed by osmotic shock and finally neutrophils are
Suspend in Hanks buffer to a concentration of 10 ⁶ cells / ml.
1 × 10 ⁵ neutrophils and the stimulant formyl-methio
10 ⁷ M of nyl-leucyl-phenylalanine (fMLP) was incubated at 37 ° C., and simultaneously the active oxygen produced by adding the compound of the present invention was measured. For measurement of active oxygen, 2-methyl-6-phenyl-3,7-dihydroimidazo
[1,2a] pyradin-3-one (CLA) reacts with active oxygen to form an excited carbonyl form, which emits light at 380 nm in the process of transition to the ground state. Was. The active oxygen production suppression rate was calculated by the following equation. Table 4 shows the results.

[0069]

(Equation 5)

[Table 4]

Example 13 In the osteoporosis condition, it is considered that the balance between bone formation and bone destruction is lost, and bone destruction is considered to be in an enhanced state. Bone destruction is believed to result from increased activation and number of osteoclasts, seeded mouse bone cells on a piece ivory as the model, there is experimental cause bone resorption in active vitamin D ₃ stimulation. Using this model, the effect of the compound of the present invention on inhibiting bone resorption was measured.

The femur and the tibia were separated from 10- to 15-day-old ICR mice, minced in an α-MEM medium containing 5% fetal bovine serum, and bone cells containing bone marrow cells and bone matrix. A suspension was prepared. Large pieces of bone were removed with a nylon mesh, live cells were stained with trypan blue exclusion, and osteoclasts were stained with tartrate-resistant acid phosphatase.
A cell suspension containing osteoclasts at a ratio of about 0.05 to 0.1% was prepared. The ivory was cut to a thickness of 150 μm with a low-speed rotating diamond cutter, and punched out with a punch according to the size of the well of the 96-well plate. These ivory pieces were placed in a 96-well plate, and the above-prepared cell suspension was added thereto with 50% osteoclasts per well.
0 was inserted. 10nM of the active form of vitamin D ₃
Was added as a stimulant, and at the same time,
And at a concentration of 100 μM. 37 ° C, 10% C
Four days after culturing the cells in an O ₂ environment, the absorption pits formed on the ivory pieces were stained with hematoxylin, observed under a microscope, and counted. The following formula was used to calculate the rate of inhibiting the formation of absorption pits.

[0072]

(Equation 6) Table 5 shows the results. The results were statistically calculated by the Student's t-test, and those with a significance level P <0.05 relative to the active vitamin D3 stimulated control group were marked with an asterisk (*) and P <0.
Significant ones with 01 are marked with **.

[0073]

[Table 5]

[0074]

The compound of the present invention has an IL-1 inhibitory action,
With antioxidant action, bone resorption inhibitory action, etc., anti-inflammatory drugs, analgesics, anti-rheumatic drugs, bone metabolism drugs, autoimmune drugs,
It is useful as an infectious agent, dermatological agent, antiallergic agent, antioxidant, and therapeutic agent for ischemic organ damage.

────────────────────────────────────────────────── ─── Continued on the front page (51) Int.Cl. ⁷ Identification code FI A61P 29/00 A61P 29/00 101 101 39/06 39/06 43/00 111 43/00 111 C07F 9/40 C07F 9/40 E (72) Inventor Takumi Uchiro 1111 Tehiro, Kamakura-shi, Kanagawa Prefecture Toray Industries, Inc. Basic Research Laboratory (56) References JP-A-3-112994 (JP, A) JP-A-49-27339 (JP, A) J . Med. Chem. , 1987, Vol. 30, No. 8, p1426-1433 Biochimica et Biophysica Acta, 1985, Vol. 818, no. 1, p9 Izv. Akad. Nauk SSS R, Ser. Khim. , 1987, No. 4, p. 860-4 Izv. Akad. Nauk SSS R, Ser. Biol. , 1980, No. 2, p289-92 (58) Fields investigated (Int. Cl. ⁷ , DB name) C07F 9/38 C07F 9/40 CA (STN) REGISTRY (STN)

Claims (9)

(57) [Claims] 1. A methylene diphosphonic acid derivative represented by the general formula (I). Embedded image [In the formula, X represents a substituent on a naphthyl group, and is a halogen, a nitro group, a nitrile group, a trifluoromethyl group, Group (provided that Z ¹ and Z ² independently represent a hydrogen atom or an alkyl group, or Z ¹ and Z ² may form a carbon ring or a carbon ring containing a heteroatom) ), Embedded image Groups (wherein Z ¹ and Z ² are the same as above and Z ³ represents oxygen or sulfur), thiol group, alkylthio group, arylthio group, acylamino group, acylthio group, acyl group, alkenyl group, aryl group , Cycloalkyl group, COOH group or COO alkyl group, m represents an integer of 0 to 3, Y represents a substituent on a naphthyl group, halogen, nitro group, nitrile group, alkyl group, alkoxy group , Trifluoromethyl group, Groups (provided that Z ¹ and Z ² are the same as above), Groups (however, Z ¹ , Z ² and Z ³ are the same as described above), thiol group, hydroxyl group, alkylthio group, arylthio group, acyloxy group, acylamino group, acylthio group, acyl group, alkenyl group, aryl group, cyclo group An alkyl group, a COOH group, or a COO alkyl group, n represents an integer of 0 to 4, m X and n Y
May be the same or different, A is-(CH
2) a-S- (a is Ri integer der of 0, A sulfur atom
Is directly bonded to methanebisphosphonic acid ), B means hydrogen, R ¹ , R ² , R ³ and R ⁴ are hydrogen, carbon atom 1
-7 linear or branched alkyl groups or pharmacologically acceptable cations, which may be the same or different. ] 2. The methylene diphosphonic acid derivative according to claim 1, wherein the naphthyl group is a 1-naphthyl group or a 2-naphthyl group. 3. A compound of the formula (IV) in the presence of a base (Wherein X, Y, m, n and a are the same as described above), and a diphosphonate compound of the general formula (II) Wherein R ′ is a linear or branched alkyl group having 1 to 7 carbon atoms, which may be the same or different.
The method for producing a methylene diphosphonic acid derivative according to claim 1, wherein a methylene diphosphonic acid derivative of the formula (I) is obtained. 4. An anti-inflammatory drug comprising the methanediphosphonic acid derivative according to claim 1 as an active ingredient. 5. An antirheumatic drug comprising the methanediphosphonic acid derivative according to claim 1 as an active ingredient. 6. A drug for a bone metabolic disease comprising the methanediphosphonic acid derivative according to claim 1 or 2 as an active ingredient. 7. An interleukin-1 inhibitor comprising the methanediphosphonic acid derivative according to claim 1 or 2 as an active ingredient. 8. An antioxidant comprising the methanediphosphonic acid derivative according to claim 1 as an active ingredient. 9. A bone resorption inhibitor comprising the methanediphosphonic acid derivative according to claim 1 as an active ingredient.