JPH08325282A - Bisphosphonic acid derivative - Google Patents

Abstract

PURPOSE: To obtain a bisphosphonic acid derivative, having a partial structure of epoxysuccinic acid and capable of manifesting inhibiting activities against cysteine proteases and providing a therapeutic agent for osteopathy and a medicine for treatment, prevention, etc., of diseases in which abnormal sthenia of cathepsins L and B participate. CONSTITUTION: The new bisphosphonic acid derivative of formula I R<1> is H, an alkyl or an aralkyl; X<1> is O or NR<6> (R<6> is the same species as that of R<1> ) or a bivalent heterocyclic ring residue; A<1> is a single bond, an amino acid residue or a dipeptide residue having the right side as the N-terminal; A<2> is a single bond, an amino acid residue or a dipeptide residue having the left side as the N-terminal; L is a bivalent connecting group; R<2> to R<5> are each is the same species as that of R<1> ), etc., having a partial structure of epoxysuccinic acid and capable of manifesting inhibiting activities against cysteine proteases and useful as a therapeutic agent for osteopathy and a therapeutic, a preventing agents, etc., for diseases in which abnormal sthenia of cathepsins L and B participate. The compound is obtained by reacting an epoxysuccinic acid derivative of formula II with a biphosphonic acid derivative of formula III in the presence of a condensing agent.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a novel bisphosphonic acid derivative and a therapeutic agent for bone diseases using the same.

[0002]

2. Description of the Related Art Bone tissue repeats bone resorption by osteoclasts and bone formation by osteoblasts, and the structure and amount of bone are maintained on this balance. However, when bone resorption becomes dominant, osteoporosis, malignant hypercalcemia,
Bone develops bone diseases such as Behcet's disease.

Bone resorption by osteoclasts can be divided into the steps of dissolution of minerals (decalcification) and decomposition of bone matrix, which is considered to be caused by lysosomal enzymes. In recent studies, it is said that the lysosomal enzymes that play a central role are cysteine protease cathepsin L and L-like enzymes (Kakegawa, Katsunuma, Molecular Medicine, 30 (10), 1310- 1318 (19
93) and Tezuka et al., J. Biol. Chem., 269 , 1106-1109.
(1994)). In addition, cysteine protease inhibitors have been reported to suppress bone resorption (JM Delaisse et al., Biochem. Biophys., Res. Commun., 125 , 441-447.
(1984)). Therefore, compounds that inhibit cysteine proteases such as cathepsin L are considered to be promising for the treatment of bone diseases such as osteoporosis.
The use of some epoxysuccinic acid derivatives for the treatment of bone disorders has been proposed (JP 63-284127).
Japanese Patent Laid-Open No. 2-218610). However, there has been no clinical use of a cysteine protease inhibitor, and its research has just started.

On the other hand, bisphosphonic acid derivatives are known to have a high affinity for bone tissue (A. Jung et al., Calcif. Tissue Res., 11 , 269-280 (1973)). Focusing on this point, an attempt was made to create a therapeutic agent for a bone disease with a reduced effect on other organs other than bone by introducing a bisphosphonate group into a compound that is confirmed or expected to be useful for bone diseases. Many have been done. As an example of the compound into which a bisphosphonate group is introduced, an estrogen which has already been used for treating osteoporosis (Japanese Patent Laid-Open No. 4-3
52795, 5-230086, 5-2869
Nos. 93 and 6-100576) and anti-inflammatory agents (Japanese Patent Laid-Open No. 2-268190).

[0005]

SUMMARY OF THE INVENTION An object of the present invention is to provide a compound and a drug which are useful for preventing or treating bone diseases such as osteoporosis, malignant hypercalcemia, and bone Behcet's disease. It is also an object of the present invention to provide compounds and agents useful as therapeutic agents for osteoarthritis and rheumatoid arthritis associated with abnormally elevated cathepsin L activity. Another object of the present invention is to provide a compound useful for treating diseases such as muscular dystrophy and muscular atrophy in which cathepsin B and L are involved.

[0006]

Means for Solving the Problems As a result of intensive studies for solving the above problems, the present inventors have found that a bisphosphonate having an osteophilic bisphosphonate group introduced into an epoxysuccinic acid derivative having a cysteine protease inhibitory action. The inventors have found that the acid derivative or its salt is useful for the prevention or treatment of bone diseases and completed the invention. The present invention has a partial structure of epoxysuccinic acid,
There is provided a bisphosphonic acid derivative or a salt thereof that exhibits ze inhibitor activity. In this bisphosphonic acid derivative, the carbonyl group of epoxysuccinic acid is preferably bonded to the bisphosphonic acid structure by a linking group. The above-mentioned bisphosphonic acid derivative has a partial structure of epoxysuccinic acid and a partial structure of a linking group which undergoes physicochemical conversion or is metabolized in vivo to release an epoxysuccinic acid derivative exhibiting cysteine protease inhibitory activity. Is also included. The present invention particularly provides a bisphosphonic acid derivative represented by the following formula (I) or a salt thereof.

[0007]

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(In the above formula, R ¹ is a hydrogen atom, an alkyl group or an aralkyl group; X ¹ is —O—,
-NR ⁶ -or a divalent heterocyclic residue; A ¹ is a single bond or an amino acid residue or dipeptide residue having the N-terminal on the right side; A ² is a single bond or a N-side on the left side.
A terminal amino acid residue or dipeptide residue;
L ¹ is a divalent linking group; and R ² , R ³ , R
⁴ , R ⁵ , and R ⁶ are each independently a hydrogen atom,
It is an alkyl group or an aralkyl group. ) The alkyl group, aralkyl group, divalent heterocyclic residue, alkylene group and phenylene group may have a substituent. In the above formula (I), L ¹ is —O—,
—NR ⁷ — (wherein R ⁷ is a hydrogen atom, an alkyl group, or an aralkyl group), a divalent heterocyclic residue, an alkylene group, —CO—, a phenylene group, or a combination thereof. It is preferably a divalent linking group.

Of these, a bisphosphonic acid derivative represented by the following formula (II) or (III) or a salt thereof is particularly effective.

[0010]

[Chemical 6]

(In the above formula, R ²¹ is an alkyl group or an aralkyl group; X ²¹ is —NR ²⁶ — or a divalent heterocyclic residue; A ²¹ is a single bond or the right side is N-terminal. A ²² is a single bond or an amino acid residue having the N-terminal on the left side; L ²¹ is —O—, —NR ²⁷ —, or a divalent heterocyclic residue. A divalent linking group selected from the group consisting of a group, an alkylene group, —CO—, a phenylene group, and combinations thereof; and R ²² , R ²³ , R ²⁴ , R ²⁵ , R ^26, and R ²⁷ are And each independently, a hydrogen atom, an alkyl group or an aralkyl group.) The above alkyl group, aralkyl group, divalent heterocyclic residue, alkylene group and phenylene group may have a substituent.

[0012]

[Chemical 7]

(In the above formula, R ³¹ is a hydrogen atom, an alkyl group or an aralkyl group; X ³¹ is —O— or —NR ³⁶ —; A ³² is a single bond or the left is an N-terminal. L ³¹ is —O—, —NR ³⁷ —, a divalent heterocyclic residue,
A divalent linking group selected from the group consisting of an alkylene group, -CO-, a phenylene group and combinations thereof;
R ³² , R ³³ , R ³⁴ , R ³⁵ , R ³⁶ and R ³⁷ are each independently a hydrogen atom, an alkyl group or an aralkyl group. The alkyl group, aralkyl group, divalent heterocyclic residue, alkylene group and phenylene group may have a substituent.

[0014]

BEST MODE FOR CARRYING OUT THE INVENTION The first aspect of the present invention is a bisphosphonic acid derivative or a salt thereof having a partial structure of epoxysuccinic acid and exhibiting cysteine protease inhibitory activity. In this bisphosphonic acid derivative, the carbonyl group of the epoxysuccinic acid is preferably bonded to the bisphosphonic acid structure by a linking group. The above-mentioned bisphosphonic acid derivative also has a partial structure of epoxysuccinic acid and a partial structure of a linking group which undergoes physicochemical conversion or is metabolized in vivo to release an epoxysuccinic acid derivative showing cysteine protease inhibitory activity. Including. The bisphosphonic acid derivative represented by the following formula (I) can be mentioned as a representative example of the bisphosphonic acid derivative having a partial structure of the above-mentioned epoxysuccinic acid and exhibiting cysteine protease inhibitory activity or a salt thereof. Taking a compound as an example,
The present invention will be described.

[0015]

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In the above formula (I), R ¹ is a hydrogen atom, an alkyl group or an aralkyl group. The number of carbon atoms of the alkyl group (when it has a substituent, the total number of carbon atoms including the substituent) is preferably 1 to 30, more preferably 1 to 20, and further preferably 1 to 1
Most preferably, it is 5. The alkyl group may have a cyclic structure, but a chain alkyl group is preferable to a cyclic alkyl group. The chain alkyl group may have a branch. These alkyl groups may have a substituent. Examples of the substituent of the alkyl group include a halogen atom and a hydroxyl group. Number of carbon atoms in aralkyl group (total number of carbon atoms including substituents, if any)
Is preferably 7 to 40, more preferably 7 to 30, and most preferably 7 to 20. Examples of aralkyl groups include benzyl, phenethyl and diphenylmethyl. Diphenylmethyl is especially preferred. The aralkyl group may have a substituent. Examples of the substituent of the aralkyl group include a halogen atom, a hydroxyl group, an alkyl group (having 1 to 10 carbon atoms) and an alkoxy group (having 1 to 10 carbon atoms).

In the formula (I), X ¹ is --O-- or --NR.
^{A 6-} or divalent heterocyclic residue is preferred. Here, R ⁶ is a hydrogen atom or an alkyl group (having 1 to 1 carbon atoms).
0) or an aralkyl group (having 7 to 20 carbon atoms). Of these, a hydrogen atom is particularly preferable. The number of carbon atoms of the alkyl group of R ⁶ (when it has a substituent, the total number of carbon atoms including the substituent) is preferably 1 to 10, more preferably 1 to 6. Through 4
Is most preferable. A chain alkyl group is preferred. The chain alkyl group may have a branch. In addition,
The alkyl group may have a substituent. Examples of the substituent of the alkyl group include a halogen atom and a hydroxyl group. The number of carbon atoms of the aralkyl group of R ⁶ (when it has a substituent, the total number of carbon atoms including the substituent) is 7 to 2
It is preferably 0, more preferably 7 to 15, and most preferably 7 to 10. Examples of aralkyl groups include benzyl and phenethyl. The aralkyl group may have a substituent. Examples of the substituent of the aralkyl group include a halogen atom, a hydroxyl group, an alkyl (having 1 to 10 carbon atoms) and an alkoxy group (having 1 to 10 carbon atoms).

The divalent heterocyclic residue preferably has a 4-membered to 7-membered heterocyclic ring, and more preferably a 5-membered or 6-membered heterocyclic ring. The heterocycle preferably has nitrogen as the heteroatom. More preferably, the heterocycle is composed solely of carbon and nitrogen. The heterocycle is preferably saturated. Particularly preferred heterocycles include a piperazine ring and a piperidine ring. If the heterocycle consists only of carbon and nitrogen,
At least one of the two free valences of the heterocyclic residue is preferably present in nitrogen. In the case of a 6-membered heterocyclic ring composed only of carbon and nitrogen, it is preferable that the free valence of the heterocyclic residue exists at the 1-position and 4-position of the ring. The heterocyclic residue may have a substituent. Examples of the substituent of the heterocyclic residue include a halogen atom, a hydroxyl group, an alkyl group, an aryl group and an alkoxy group as described above.

In the above formula (I), A ¹ is a single bond,
Alternatively, it is an amino acid residue or dipeptide residue having the N-terminal on the right side. In the case of a single bond, the above X ¹ is directly bonded to carbonyl (carbonyl bonded to the oxirane ring). Amino acid residues and dipeptide residues are preferably derived from naturally occurring L-type amino acids. Glycine, alanine, valine, leucine, isoleucine, phenylalanine or tyrosine residues or combinations thereof are especially preferred. In the case of dipeptide residues, different amino acid residues may be combined. The two carbons of the oxirane ring shown in the above formula (I) are both asymmetric carbon atoms. Formula (I) shows that the two carbonyl groups attached to the oxirane ring are trans. That is, the bisphosphonic acid derivative of the present invention is any one of the following optical isomers (T1) or (T2), or a mixture thereof. Among them, the optical isomer of (T1) is preferable.

[0020]

[Chemical 9]

In the above formula (I), A ² is a single bond or an amino acid residue or dipeptide residue having the N-terminal on the left side. In the case of a single bond, L ¹ described below is directly bonded to carbonyl (carbonyl bonded to the oxirane ring). Amino acid residues and dipeptide residues are preferably derived from naturally occurring L-type amino acids. Particularly preferred are residues of glycine, alanine, valine, leucine, isoleucine, phenylalanine or tyrosine, or a combination thereof. In the case of dipeptide residues, different amino acid residues may be combined.

In the formula (I), L ¹ is --O-- or --N.
R ⁷ -, a divalent heterocyclic residues, alkylene, -CO-, is preferably phenylene and divalent linking group selected from the group consisting of. R ⁷ above
Is a hydrogen atom, an alkyl group or an aralkyl group.
Of these, a hydrogen atom is particularly preferable. The number of carbon atoms of the alkyl group of R ⁷ (when it has a substituent, the total number of carbon atoms including the substituent) is preferably from 1 to 10, more preferably from 1 to 6. Most preferably, it is from 4 to 4. A chain alkyl group is preferred.
The chain alkyl group may have a branch. The alkyl group may have a substituent. Examples of the substituent of the alkyl group include a halogen atom and a hydroxyl group. The number of carbon atoms of the aralkyl group of R ⁷ (when it has a substituent, the total number of carbon atoms including the substituent) is 7 to 2
It is preferably 0, more preferably 7 to 15, and most preferably 7 to 10. Examples of this aralkyl group include benzyl and phenethyl. The aralkyl group may have a substituent. Examples of the substituent of the aralkyl group include a halogen atom, a hydroxyl group, an alkyl group and an alkoxy group as described above.

The divalent heterocyclic residue preferably has a 4-membered to 7-membered heterocyclic ring, more preferably a 5-membered or 6-membered heterocyclic ring. The heterocycle preferably has nitrogen as the heteroatom. More preferably, the heterocycle is composed solely of carbon and nitrogen. The heterocycle is preferably saturated. Particularly preferred heterocycles include a piperazine ring and a piperidine ring. If the heterocycle consists only of carbon and nitrogen,
At least one of the two free valences of the heterocyclic residue is preferably present in nitrogen. In the case of a 6-membered heterocyclic ring composed only of carbon and nitrogen, it is preferable that the free valence of the heterocyclic residue exists at the 1-position and 4-position of the ring. The heterocyclic residue may have a substituent. Examples of the substituent of the heterocyclic residue include a halogen atom, a hydroxyl group, an alkyl group, an aryl group and an alkoxy group as described above.

The number of carbon atoms of the alkylene group (the total number of carbon atoms including the substituents in the case of having a substituent) is preferably from 1 to 30, more preferably from 1 to 20. Most preferably, it is from 15 to 15.
The alkylene group may have a cyclic structure, but a chain alkylene group is preferable to a cyclic alkylene group. The chain alkylene group may have a branch. The alkylene group may have a substituent. Examples of the substituent of the alkylene group include a halogen atom, hydroxyl and aryl as described above. The phenylene group may have a substituent. Examples of the substituent of phenylene include halogen atom, hydroxyl, alkyl and alkoxy as described above.

Examples of L ¹ are shown in L ^{2 to} L ¹⁵ below. L ^2: - heterocyclic residue - L ^3: - heterocyclic residue - alkylene - L ^4: -O- heterocyclic residue - L ^5: -O- alkylene ^{-CO-NR 8 - L 6:} -NR 9 - alkylene ^{-CO-NR 10 - L 7:} -O- alkylene - L ^8: - heterocyclic residue -CO- alkylene - L ^9: - heterocyclic residue -CO-O- alkylene - L ^10: - heterocycle residues -O-CO- alkylene - L ^11: -NR ¹¹ - alkylene -NR ¹² -CO- alkylene - L ^12: - heterocyclic residue - alkylene - phenylene -O- alkylene - L ^13: - heterocyclic residue -Alkylene-phenylene-OC
O- alkylene - L ^14: - heterocyclic residue - alkylene - phenylene -O- alkylene ^{-CO-NR 13 - L 15:} -NR 14 - alkylene -O-CO- alkylene -

In each of the above formulas, R ⁸ , R ⁹ , R ¹⁰ , R
¹¹ , R ¹² , R ¹³ and R ¹⁴ are each a hydrogen atom, an alkyl group or an aralkyl group. A hydrogen atom is particularly preferred. The details of the alkyl group and aralkyl group are the same as those of R ⁷ described above. Further, the details of the heterocyclic residue, the alkylene group and the phenylene group are the same as those described above for L ¹ .

In the formula (I), R ² , R ³ , R ⁴ and R ⁵ are each independently a hydrogen atom, an alkyl group or an aralkyl group. A hydrogen atom is particularly preferred. R
^The number of carbon atoms of the alkyl group of ^{2 to} R ⁵ (when it has a substituent, the total number of carbon atoms including the substituent) is 1 to 10
Is preferable, 1 to 6 is more preferable, and 1 to 4 is most preferable. A chain alkyl group is preferred. The chain alkyl group may have a branch. The number of carbon atoms of the aralkyl group of R ^{2 to} R ⁵ (when it has a substituent, the total number of carbon atoms including the substituent)
Is preferably 7 to 20, more preferably 7 to 15, and most preferably 7 to 10. Examples of aralkyl groups include benzyl and phenethyl. The aralkyl group may have a substituent. Examples of the aralkyl substituent include a halogen atom, a hydroxyl group, an alkyl group and an alkoxy group as described above.

Examples of preferable bisphosphonic acid derivatives are represented by the following formula (II).

[0029]

[Chemical 10]

In the formula (II), R ²¹ is an alkyl group or an aralkyl group. Aralkyl groups are particularly preferred. For details of these alkyl groups and aralkyl groups, see
It is the same as R ^{1 of the} above formula (I). In the formula (II), X ²¹ is —NR ²⁶ — or a divalent heterocyclic residue. R ²⁶ is a hydrogen atom, an alkyl group or an aralkyl group. A hydrogen atom is particularly preferred. For details of these alkyl groups and aralkyl groups, refer to R ^{6 of the} above formula (I).
Is the same as The details of the divalent heterocyclic residue are also the same as those of X ^{1 in} the above formula (I).

In the formula (II), A ²¹ is a single bond or an amino acid residue or dipeptide residue having the N-terminal on the right side. In the case of a single bond, the above X ²¹ is directly bonded to the carbonyl group (carbonyl bonded to the oxirane ring). The details of the amino acid residue and the dipeptide residue are the same as those of A ¹ of the formula (I) described above. The formula (II) shows that, like the formula (I), the two carbonyl groups bonded to the oxirane ring are in the trans form. In the formula (II), A ²² is a single bond or an amino acid residue having the left side as the N-terminal. In the case of a single bond, the following L ²¹ is directly bonded to the carbonyl group (carbonyl bonded to the oxirane ring). The details of the amino acid residue are the same as those of A ^{2 in} formula (I) described above.

In the formula (II), L ²¹ is --O-- or --N.
R ²⁷ -, a divalent heterocyclic residue, an alkylene group, -CO-,
It is a divalent linking group selected from the group consisting of phenylene groups and combinations thereof. Among them, -O-, -N
R ²⁷ -, a divalent heterocyclic residue, an alkylene group, -CO- and is preferably selected from the group consisting of. R ²⁷ is a hydrogen atom, an alkyl group or an aralkyl group. A hydrogen atom is particularly preferred. The details of the alkyl group and aralkyl group of R ²⁷ are the same as those of R ^{7 of} formula (I) described above. The details of the above divalent heterocyclic residue, alkylene group and phenylene group are also the same as those for L ^{1 in} formula (I) described above.

Examples of L ²¹ are shown in L ^{22 to} L ²⁶ below. L ^22: - heterocyclic residue - L ^23: -O- heterocyclic residue - L ^24: -O- alkylene ^{-CO-NR 28 - L 25:} -NR 29 - alkylene -CO-NR ³⁰ - L ^26: -O- alkylene - in the above formulas, R ^28, R ²⁹ and R ³⁰ are each a hydrogen atom, an alkyl group or an aralkyl group.
A hydrogen atom is particularly preferred. The details of the alkyl group and the aralkyl group are the same as those of R ^{7 in} formula (I) described above. Further, the details of the heterocyclic residue, the alkylene group, and the phenylene group are also the same as L ^{1 of} the above formula (I).

In the formula (II), R ²² , R ²³ , R ²⁴ and R ²⁵ are each a hydrogen atom, an alkyl group or an aralkyl group. A hydrogen atom is particularly preferred. For details of the alkyl group and the aralkyl group, refer to R ^{2 of} the formula (I) described above.
Is the same as ~ R ⁵ .

Another preferred bisphosphonic acid derivative is represented by the following formula (III).

[0036]

[Chemical 11]

In the above formula (III), R ³¹ is a hydrogen atom, an alkyl group or an aralkyl group. A hydrogen atom or an alkyl group is preferred. The details of the alkyl group and the aralkyl group are the same as R ¹ of the above formula (I).
In the formula (III), X ³¹ is —O— or —NR ³⁶ —. -O- is particularly preferable. R ³⁶ is a hydrogen atom,
It is an alkyl group or an aralkyl group. A hydrogen atom is particularly preferred. The details of the alkyl group and the aralkyl group are the same as those of R ^{6 in} formula (I) described above. The formula (III) shows that, like the formula (I), the two carbonyl groups bonded to the oxirane ring are in the trans form. In the formula (III), A ³² is a single bond or an amino acid residue or a dipeptide residue having the N-terminal on the left side. In the case of a single bond, the following L ³¹ is directly bonded to carbonyl (two carbonyl groups bonded to the oxirane ring). The details of the amino acid residue and the dipeptide residue are the same as those of A ^{2 in} formula (I) described above.

In the formula (III), L ³¹ is --O-- or --N.
R ^37- , a divalent heterocyclic residue, an alkylene group, -CO-,
It is a divalent linking group selected from the group consisting of phenylene groups and combinations thereof. R ³⁷ is a hydrogen atom,
It is an alkyl group or an aralkyl group. A hydrogen atom is particularly preferred. The details of the alkyl group and aralkyl group of R ³⁷ are the same as those of R ⁷ of the above formula (I). The details of the above divalent heterocyclic residue, alkylene group and phenylene group are also the same as those for L ^{1 in} formula (I) described above.

Examples of L ³¹ are shown in L ^{32 to} L ⁴¹ below. L ³² : -heterocyclic residue-L ³³ : -heterocyclic residue-alkylene- L ³⁴ : -heterocyclic residue-CO-alkylene-L ³⁵ : -heterocyclic residue-CO-O-alkylene-L ³⁶ : - heterocyclic residue -O-CO- alkylene - L ^37: -NR ³⁸ - alkylene -NR ³⁹ -CO- alkylene - L ^38: - heterocyclic residue - alkylene - phenylene -O- alkylene - L ^39: - Heterocyclic residue-alkylene-phenylene-OC
O- alkylene - L ^40: - heterocyclic residue - alkylene - phenylene -O- alkylene ^{-CO-NR 40 - L 41:} -NR 41 - alkylene -O-CO- alkylene -

In the above formulas, R ³⁸ , R ³⁹ , R ⁴⁰ and R ⁴¹ are each a hydrogen atom, an alkyl group or an aralkyl group. A hydrogen atom is particularly preferred. The details of the alkyl group and the aralkyl group are the same as those of R ^{7 in} formula (I) described above. Further, the details of the heterocyclic residue, the alkylene group, and the phenylene group are also the same as L ^{1 of} the above formula (I). In the formula (III), R ³² ,
R ³³ , R ³⁴ and R ³⁵ are each a hydrogen atom, an alkyl group or an aralkyl group. A hydrogen atom is particularly preferred. The details of the alkyl group and the aralkyl group are the same as those of R ^{2 to} R ⁵ in the formula (I) described above.

Specific examples of the bisphosphonic acid derivative of the present invention are shown below.

[0042]

[Chemical 12]

[0043]

[Chemical 13]

[0044]

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[0045]

[Chemical 15]

[0046]

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[0047]

[Chemical 17]

[0048]

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[0049]

[Chemical 19]

[0050]

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[0051]

[Chemical 21]

[0052]

[Chemical formula 22]

[0053]

[Chemical formula 23]

[0054]

[Chemical formula 24]

[0055]

[Chemical 25]

[0056]

[Chemical formula 26]

[0057]

[Chemical 27]

[0058]

[Chemical 28]

[0059]

[Chemical 29]

[0060]

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[0061]

[Chemical 31]

[0062]

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[0063]

[Chemical 33]

[0064]

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[0065]

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[0066]

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[0067]

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[0068]

[Chemical 38]

[0069]

[Chemical Formula 39]

[0070]

[Chemical 40]

[0071]

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[0072]

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[0073]

[Chemical 43]

[0074]

[Chemical 44]

[0075]

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[0076]

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[0077]

[Chemical 47]

[0078]

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[0079]

[Chemical 49]

[0080]

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[0081]

[Chemical 51]

[0082]

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[0083]

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In each of the above formulae, Me is methyl, Et
Represents ethyl and Bn represents benzyl. The bisphosphonic acid derivative of the present invention may be used as a salt. When forming a salt, bisphosphonic acid and a base may form a salt, or a heterocyclic residue or an amino group may form a salt with an acid. Bisphosphonic acids may form partial salts (mono-, di- or tri-). Examples of base cations include alkali metal ions (eg, sodium, potassium) and alkaline earth metal ions (eg, calcium). Acids include physiologically acceptable inorganic acids (eg hydrochloric acid, sulfuric acid) and organic acids (eg acetic acid)
Is available. A typical method for synthesizing the bisphosphonic acid derivative of the present invention will be described. The bisphosphon derivative of the above formula (I) can be synthesized by reacting the following compound (A) or (B) with the compound (C). The compound (A) means that A ^{2 in the} formula (I) is a single bond.

[0085]

[Chemical 54]

In the above formula, R ¹ , X ¹ , A ¹ , A
² , L ¹ , R ² , R ³ , R ⁴ and R ⁵ have the same definitions as in formula (I) described above. A ³ is an amino acid residue or dipeptide residue which left the N-terminus. The above reaction is carried out in the presence of a condensing agent. Examples of the condensing agent include N, N'-dicyclohexylcarbodiimide, N-
Examples include hydroxysuccinimide / N, N′-dicyclohexylcarbodiimide, 1-hydroxybenzotriazole / 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride and N, N′-dicyclohexylcarbodiimide / 4-dimethylaminopyridine. be able to. In the above reaction, the compound (C)
The terminal atom on the left side of L ¹ of is oxygen or nitrogen. In the case of oxygen, the compound (C) is an alcohol, and an ester is produced by the reaction with the carboxyl group of the compound (A) or (B). In the case of nitrogen, the compound (C) is an amine compound, and reacts with the carboxyl group of the compound (A) or (B) to form an amide. The bisphosphonic acid derivative (I) can also be synthesized by reacting the following compound (D) with the compound (E).

[0087]

[Chemical 55]

In the above formula, R ¹ , X ¹ , A ¹ , A
² , L ¹ , R ² , R ³ , R ⁴ and R ⁵ have the same definitions as in formula (I) described above. L ¹⁶ is a divalent linking group whose left and right terminal atoms are oxygen or nitrogen, and when the right terminal atom of L ¹⁶ is oxygen, compound (D) is an alcohol or carboxylic acid. When the atom on the right side of L ¹⁶ is nitrogen, compound (D) is an amine body. Y is a halogen, hydroxyl, carboxyl or amino group. L ¹⁷ is a divalent linking group necessary for forming L ¹ by bonding with L ¹⁶ .

In the bisphosphonic acid derivative (I) of the present invention, when X ¹ is —O—, after carrying out the above reaction or the reaction with potassium hydroxide in ethanol and then acidification. R ¹ can be hydrogenated by the method described above or by catalytic reduction (when R ¹ is benzyl).

[0090]

[Chemical 56]

When R ² , R ³ , R ⁴ and R ⁵ are not hydrogen, after the above reaction or is carried out, R ² , R ³ , R ⁴ and R ⁵ are converted to hydrogen as follows. can do.

[0092]

[Chemical 57]

Furthermore, the disodium salt of bisphosphonic acid is prepared by adding a sodium chlorinating agent (eg sodium hydrogen carbonate, sodium acetate) to the bisphosphonic acid derivative (I) in which R ² , R ³ , R ⁴ and R ⁵ are hydrogen. It can be obtained by acting. At this time, when X ¹ is —O— and R ¹ is hydrogen, the carboxylic acid is also obtained as a sodium salt. The compounds (A), (B) and (D) are described in JP-A Nos. 55-115878 and 55-47.
It can be synthesized by a method similar to the method described in each of No. 668 and No. 57-169478. The specific reaction is shown below.

[0094]

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[0095]

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[0096]

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In the above formula, R ¹ , X ¹ , A ¹ , A
² , L ¹ , R ² , R ³ , R ⁴ and R ⁵ have the same definitions as in the above formula (I). R ¹⁵ and R ¹⁶ are protecting groups, specifically alkyl or aralkyl. Examples of binders used here are the same as the binders described above.

The pharmacological action of the bisphosphonic acid derivative of the present invention will be described. The bisphosphonic acid derivative of the present invention is
It has a chemical structure in which a bisphosphonate group having bone affinity is introduced into an epoxysuccinic acid derivative having a thiol protease inhibitory action. The thiol protease includes cathepsin L or B or calpain. Therefore, the bisphosphonic acid derivative of the present invention and its physiologically acceptable salt can be expected to have a pharmacological action against diseases associated with these proteases.

Diseases in which cathepsin L is involved include bone diseases such as osteoporosis, malignant hypercalcemia, and bone Behcet's disease, as described in the prior art. Therefore, the bisphosphonic acid derivative of the present invention and its physiologically acceptable salt are useful as prophylactic or therapeutic agents for these bone diseases. It has been reported that cathepsin L degrades type II, type IX and type XI of collagen constituting articular cartilage in the neutral region (FEBS Lett. 269 , 189-19.
3 (1990)). Therefore, the bisphosphonic acid derivative of the present invention and a physiologically acceptable salt thereof can be expected to be effective also for bone joint inflammation or rheumatoid arthritis, which is a bone disease accompanied by abnormally increased cathepsin L activity (special feature). (See Kaihei 5-178757). The compound of the present invention also exhibits an excellent action as a cathepsin B inhibitor.
Further, as diseases involving thiol proteases other than cathepsin L such as cathepsin B, muscular dystrophy and muscular atrophy (cathepsin B and calpain are involved),
Alzheimer's disease (involves calpain), diseases thought to be caused by demyelination of nerve cells, such as multiple sclerosis and neuropathies of the peripheral nerve (involves calpain), cataracts (involves calpain), allergic diseases (including thiol protease). Involvement), fulminant hepatitis (involves calpain), breast cancer, prostate cancer and benign prostatic hyperplasia (involves calpain), cancer growth and metastasis (involves cathepsin B and calpain), or platelet aggregation (involves calpain). (See Japanese Patent Laid-Open No. 6-239835). Therefore, the compound of the present invention is considered to be effective in treating or preventing these diseases. The bisphosphonic acid derivative of the present invention and a physiologically acceptable salt thereof can be expected to be useful as prophylactic or therapeutic agents for the above diseases, and particularly bone diseases such as osteoporosis, malignant calcemia, and bone Behçet's disease. It is useful as a preventive or therapeutic drug. The bisphosphonic acid derivative of the present invention may be administered either orally or parenterally. Examples of the dosage form of the orally-administered agent include tablets, capsules, powders, granules and syrups.
Parenteral administration methods include mucosal administration, body surface administration, vascular administration, and intratissue administration. For mucosal administration, it is used as an eye drop, an inhalant, a spray or a suppository. For body surface administration, use as an ointment. In the case of vascular administration and tissue administration, it is used as an injection. The above-mentioned oral administration preparation can be produced by using usual excipients, disintegrating agents, binders, lubricants, dyes and diluents. Glucose and lactose are generally used as excipients. Examples of disintegrants include starch and carboxymethyl cellulose calcium. Lubricants include magnesium stearate and talc. As a binder,
Hydroxypropyl cellulose, gelatin and polyvinyl alcohol are used. The parenteral preparation can also be produced by a usual method. For example, in the case of an injection, ordinary distilled water for injection, physiological saline or Ringer's solution may be used. The dose of the bisphosphonic acid derivative of the present invention is usually 0.01 to 100 mg as an injection and 0.1 to 1 g as an oral administration in an adult in general. Of course, the dose may be increased or decreased according to age, race, symptoms, etc.

[0100]

【Example】

Example 1 [4-[(2S, 3S) -3-Diphenylmethylcarbamoyloxirane-2-carbonyl] -1-piperazinyl] methylenebisphosphonate tetrabenzyl Compound Example (29)

(1-1) Synthesis of 1- (t-butoxycarbonyl) -4-formylpiperazine 1-formylpiperazine (3.42 g, 30.0 mmol) was dissolved in tetrahydrofuran (15 mL) and water (15 m).
L) and dissolved in a mixed solvent with triethylamine (3.64 g, 36.0 mmol) under ice cooling, and then di-t-butyl dicarbonate (7.85 g, 36.0 mmol) for 20 minutes. It dripped over. After stirring for 1.5 hours under ice cooling,
Tetrahydrofuran was distilled off under reduced pressure, and ethyl acetate (45 mL) was added to the residue. The organic layer was separated, washed successively with 10% aqueous citric acid solution, saturated aqueous sodium hydrogen carbonate solution and saturated brine, and dried over anhydrous sodium sulfate. After evaporating the solvent under reduced pressure, n-hexane (30 mL) was added to the obtained white solid, and the mixture was vigorously stirred at room temperature for 30 minutes, filtered, washed with n-hexane, and dried to give the title compound as a white powder. (5.24 g). ¹ H NMR (400 MHz, CDCl ₃ ) δ 1.48 (9 H, s) 3.3 to 3.6 (8 H, m) 8.08 (1 H, s)

(1-2) Synthesis of tetrabenzyl [4- (t-butoxycarbonyl) -1-piperazinyl] methylenebisphosphonate The compound (1.07 g, 5.0 mmol) obtained in (1-1) above was synthesized. Anhydrous tetrahydrofuran (THF) (10m
L) solution was cooled to −5 ° C. and oxalyl chloride (0.
(43 mL, 5.0 mmol) was added dropwise, and the mixture was stirred at room temperature for 4 hours. The reaction mixture was cooled to −10 ° C. and the suspension of the sodium salt of dibenzyl phosphite in tetrahydrofuran [6
Dibenzyl phosphite (4.2 g, 16 mmol) was added dropwise to a suspension of 0% sodium hydride (0.50 g, 12.5 mmol) in anhydrous THF (8 mL) at room temperature, and the mixture was stirred at room temperature for 3 hours. Preparation] was added dropwise so that the internal temperature did not exceed 5 ° C. After stirring at −10 ° C. for 0.5 hour and then at room temperature for 1 hour, the reaction mixture was concentrated under reduced pressure, and ethyl acetate and water were added to the residue. The organic layer was separated, the aqueous layer was extracted with ethyl acetate, the organic layers were combined, washed with saturated brine, and dried over anhydrous sodium sulfate. After evaporating the solvent under reduced pressure, the residue was purified by medium pressure silica gel chromatography (n-hexane / ethyl acetate = 1/2) to give the title compound as a white crystalline powder (597 mg). ¹ H NMR (400 MHz, CDCl ₃ ) δ 1.43 (9H, s) 2.95 (4H, m) 3.29 (4H, m) 3.43 (1H, t, J = 25 Hz) 5.0- 5.1 (8H, m) 7.2-7.4 (20Hz, m)

(1-3) Synthesis of (1-piperazinyl) methylenebisphosphonic acid tetrabenzyl p-toluenesulfonate salt (0 mL of the compound (2.16 g, 3.00 mmol) obtained in (1-2) above) ) P-Toluenesulfonic acid monohydrate (1.14 g, 6.0 mmol) was added to the suspension, and the mixture was stirred at room temperature. After 3 days, the extrudate was collected by filtration, washed several times with ether and dried under reduced pressure, and the title compound was mixed with its mono-debenzylated product [(1-piperazinyl) methylenebisphosphonic acid tribenzyl p-toluenesulfonate] ( 66:34) (white powder, 1.60, content of title compound: 1.11 g, content of monodebenzylated product; 0.48 g). FAB MS (M / z, relative intensity) 621 ([M + H] ⁺ , title compound (free base), 2
2) 531 ([M + H] ⁺ , mono-debenzylated product (free base), 7) 359 ([M-PO ₃ Bn ₂ ] ⁺ , title compound (free base), 94) 269 ([M-PO ₃ Bn ₂ ] ⁺ , Mono-debenzylated product (free base), 32)

(1-4) Synthesis of ethyl (2S, 3S) -3-diphenylmethylcarbamoyloxirane-2-carboxylate Under a nitrogen atmosphere, potassium (2S, 3S) -3-ethoxycarbonyloxirane-2-carboxylate ( 566 mg,
2.0 mmol) was suspended in dry ether (7 mL), and under ice cooling, oxalyl chloride (0.19 mL, 2.
A solution of 2 mmol) in dry ether (2 mL) was added dropwise. After stirring at room temperature for 1 hour, a solution of aminodiphenylmethane (0.69 mL, 4.0 mmol) in dry ether (2 mL) was added dropwise under ice cooling, and the mixture was further stirred at room temperature for 1 hour. The precipitate was filtered off and the residue was washed with ether. The filtrate and washings were combined, washed with saturated saline, and dried over anhydrous sodium sulfate. After evaporating the solvent under reduced pressure, the residue was purified by medium pressure silica gel column chromatography (hexane / ethyl acetate = 4/1) to give the title compound as a white crystalline powder (222 mg). ¹ H NMR (400 MHz, CDCl ₃ ) δ 1.32 (3 H, t, J = 7 Hz) 3.51 (1 H, d, J = 2 Hz) 3.77 (1 H, d, J = 2 Hz) 4.2 4.3 (2H, m) 6.22 (1H, d, J = 8Hz) 6.67 (1H, d, J = 8Hz) 7.1-7.4 (10H, m)

(1-5) Synthesis of (2S, 3S) -3-diphenylmethylcarbamoyloxirane-2-carboxylic acid (2S, 3S) -3-diphenylmethylcarbamoyloxirane-2 obtained in (1-4) above. -Ethyl carboxylate (5.50 g, 16.9 mmol) in ethanol (30
0 mL) and 0.5N potassium hydroxide / under ice cooling
An ethanol solution (40.4 mL, 20.2 mmol) was added, and the mixture was stirred at room temperature for 3 hours. The solvent was distilled off under reduced pressure, water (300 mL) was added, and then ether (100 mL x
It was washed in 2). The aqueous layer was adjusted to pH 1-2 with 2N hydrochloric acid, and extracted with ethyl acetate (100 mL × 3). The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate.
The solvent was evaporated under reduced pressure to give the title compound as a white crystalline powder (4.76 g). ¹ H NMR (400 MHz, CD ₃ OD) δ 3.57 (1 H, d, J = 2 Hz) 3.71 (1 H, d, J = 2 Hz) 6.23 (1 H, s) 7.2 to 7.4 (10H, m)

(1-6) [4-[(2S, 3S) -3-
Synthesis of tetrabenzyl diphenylmethylcarbamoyloxirane-2-carbonyl] -1-piperazinyl] methylenebisphosphonate (2S, 3S) -3-diphenylmethylcarbamoyloxirane-2-carboxylic acid (29 obtained in (1-5) above.
A suspension of 8 mg, 1.00 mmol) and N-hydroxysuccinimide (116 mg, 1.01 mmol) in methylene chloride (8 mL) was added to N, N′-dicyclohexylcarbodiimide (212 mg, 1.03) under ice cooling. Mmol) was added and the mixture was stirred at 5 ° C. After 1 hour, the above (1-
(1-piperazinyl) methylenebisphosphonate tetrabenzyl p-toluenesulfonate obtained in 3) and (1-
Piperazinyl) methylenebisphosphonate tribenzyl
Mixture of p-toluenesulfonate (762 mg, former compound content: 523 mg, 0.66 mmol, latter compound content: 239 mg, 0.34 mmol) and N-
Methylmorpholine (113 mg, 1.11 mmol) was added, and the mixture was stirred at the same temperature for 1.5 hours and further at room temperature for 4 hours. The reaction mixture was concentrated under reduced pressure, ethyl acetate and water were added to the residue, and the insoluble material (DC-Urea) was filtered off. The organic layer was separated from the filtrate, washed successively with 10% aqueous citric acid solution, saturated aqueous sodium hydrogen carbonate and saturated brine, and dried over anhydrous sodium sulfate. After evaporating the solvent under reduced pressure, the residue was subjected to medium pressure silica gel column chromatography (chloroform / methanol = 25).
/ 1) to give the title compound as a white powder (3
98 mg). ¹ H NMR (400 MHz, CDCl ₃ ) δ 2.9 to 3.0 (4 H, m) 3.2 to 3.5 (4 H, m) 3.36 (1 H, t, J = 25 Hz) 3.55 ( 1H, d, J = 2Hz) 3.65 (1H, d, J = 2Hz) 4.9-5.1 (8H, m) 6.24 (1H, d, J = 8Hz) 6.83 (1H, d, J = 8 Hz) 7.2-7.4 (30 H, m) FAB MS (M / z, relative intensity) 900 ([M + H] ⁺ , 9)) 638 ([M-PO ₃ Bn ₂ ] ⁺ , 29)

Example 2 [4-[(2S, 3S) -3-Diphenylmethylcarbamoyloxirane-2-carbonyl] -1-piperazinyl] methylenebisphosphonate disodium Disodium salt of compound example (1) Example 1 The compound obtained in (1-6) (200 mg, 0.
222 mmol) in ethyl acetate (4 mL) was added to water (4 mL).
mL) and 10% palladium / carbon (30 mg) were added, and catalytic hydrogenation was carried out at room temperature and 1 atm for 1 hour. Further sodium hydrogen carbonate (37.3 mg, 0.444 mmol)
Was added and catalytic hydrogenation was carried out for 2 hours, and then the catalyst was filtered off. The aqueous layer was separated and the water was evaporated under reduced pressure to give the title compound as a pale brown powder (130 mg). ¹ H NMR (400 MHz, D ₂ O) δ 3.39 (1H, t, J = 18 Hz) 3.6 to 3.8 (4H, m) 3.82 (1H, d, J = 2 Hz) 3.8 -4.0 (4H, m) 4.15 (1H, d, J = 2Hz) 6.19 (1H, s) 7.3-7.5 (10H, m) FAB MS (m / z, relative intensity) ) 606 ([M + Na] ⁺ , 9) 584 ([M + H] ⁺ , 7) IR (KBr) cm ⁻¹ : 3408, 3304, 306
2,3030, 1655, 1543, 1495, 145
2,1246,1155,1092,901,700,
540,463

Example 3 [4- [N-[(2S, 3S) -3-diphenylmethylcarbamoyloxirane-2-carbonyl] glycyl]
Oxypiperidino] methylenebisphosphonate tetrabenzyl Compound example (34) (3-1) Synthesis of 1-formyl-4-piperidone ethylene ketal 98% in acetic anhydride (51.0 mL, 0.54 mmol)
Formic acid (21.5 mL, 0.57 mmol) was added and 50
Heated at ~ 60 ° C for 2 hours. After cooling to room temperature, 4-piperidone ethylene ketal (36.0 g, 0.
25 mmol) was added dropwise so that the internal temperature did not exceed 30 ° C. Stirring was continued at room temperature for 1 hour, then ice water (250 mL) was added to the reaction mixture and the mixture was stirred for a while.
Aqueous sodium hydroxide solution was added dropwise to adjust the pH to 5 to 6, and sodium chloride was added until saturation. It was extracted with ethyl acetate (150 mL × 4), and the extracts were combined and saturated aqueous potassium carbonate solution (80
It was washed with mL × 2) and saturated saline (80 mL), and dried over anhydrous sodium sulfate. After evaporating the solvent under reduced pressure, the title compound was obtained as a colorless oil (33.8 g). ¹ H NMR (400 MHz, CDCl ₃ ) δ 1.6 to 1.7 (4H, m) 3.45 (2H, m) 3.62 (2H, m) 3.99 (4H, s) 8.03 ( 1H, s)

Example 3 (3-2) (1,4-dioxa-8-azaspiro [4.
5] Synthesis of tetrabenzyl decan-8-yl) methylenebisphosphonate A solution of the compound (2.81 g, 16.4 mmol) obtained in (3-1) above in anhydrous THF (16 mL) was added to -5 to -10.
Cooled to ℃, oxalyl chloride (1.41 mL, 1
(6.4 mmol) was added dropwise, and the mixture was stirred at the same temperature for 1 hour and further at room temperature for 4 hours. The reaction mixture was cooled again to −10 ° C., and a suspension of sodium salt of dibenzyl phosphite in tetrahydrofuran (25 mL) [dibenzyl phosphite (1
3.8 g, 52.5 mmol) and 60% sodium hydride (1.64 g, 41 mmol)] with an internal temperature of 5
It was added dropwise so as not to exceed ℃. 0.5 at -5 to -10 ° C
After stirring for 1 hour and then at room temperature for 1 hour, the reaction mixture was concentrated under reduced pressure and ethyl acetate and water were added to the residue. The organic layer was separated, the aqueous layer was extracted with ethyl acetate, the organic layers were combined, washed with saturated brine, and dried over anhydrous sodium sulfate. After evaporating the solvent under reduced pressure, the residue was purified by medium pressure silica gel column chromatography (n-hexane / ethyl acetate = 1/5) to obtain the title compound as a white crystalline powder (2.
48 g). ¹ H NMR (400 MHz, CDCl ₃ ) δ 1.63 (4H, m) 3.11 (4H, m) 3.53 (1H, t, J = 25 Hz) 3.91 (4H, s) 5.0- 5.1 (8H, m) 7.2-7.4 (20H, m)

(3-3) Synthesis of tetrabenzyl (4-oxopiperidino) methylenebisphosphonate A solution of the compound (2.04 g, 3.01 mmol) obtained in (3-2) above in acetone (30 mL) was added to pyridinetosylic acid. Salt (226 mg, 0.90 mmol) and water (3 m
L) was added and the mixture was heated under reflux for 48 hours. After distilling off the solvent under reduced pressure,
The residue was dissolved in ethyl acetate, washed with saturated aqueous sodium hydrogen carbonate and saturated brine, and dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure to give the title compound as a yellow oil (2.04 g). ¹ H NMR (400 MHz, CDCl ₃ ) δ 2.23 (4H, m) 3.22 (4H, m) 3.53 (1H, t, J = 25 Hz) 5.0 to 5.1 (8H, m) 7.2-7.4 (20H, m)

(3-4) (4-hydroxypiperidino)
Synthesis of tetrabenzyl methylenebisphosphonate To a solution of the compound (2.04 g) obtained in (3-3) above in methanol (10 mL) was added sodium borohydride (114 mg, 3.01 mmol) under ice cooling, and the mixture was cooled with ice. 2 below
Stirred for 0 minutes. The solvent was evaporated under reduced pressure, ethyl acetate and 10% aqueous ammonium chloride solution were added to the residue, and the organic layer was separated and washed with saturated aqueous sodium hydrogen carbonate and saturated brine. After drying over anhydrous sodium sulfate and evaporating the solvent under reduced pressure, the obtained white solid was purified by medium pressure silica gel calm chromatography (ethyl acetate) to obtain the title compound as a white crystalline powder (1.41 g). ¹ H NMR (400 MHz, CDCl ₃ ) δ 1.4 to 1.5 (2H, m) 1.7 to 1.8 (2H, m) 1.6 (1H, br) 2.9 to 3.0 ( 2H, m) 3.1-3.2 (2H, m) 3.48 (1H, t, J = 25Hz) 3.59 (1H, m) 5.0-5.1 (8H, m) 7. 2-7.3 (20H, m)

(3-5) Synthesis of N-[(2S, 3S) -3-diphenylmethylcarbamoyloxirane-2-carbonyl] glycine ethyl ester Obtained in (1-5) of Example 1 (2S, 3S). A suspension of 3-diphenylmethylcarbamoyloxirane-2-carboxylic acid (595 mg, 2.00 mmol) and N-hydroxysuccinimide (24 g mg, 2.21 mmol) in methylene chloride (12 mL) was cooled with ice. DCC (455
mg, 2.21 mmol) was added and the mixture was stirred at 5 ° C. Three
After 0 minutes, glycine ethyl ester hydrochloride (307 m
g, 2.20 mmol) and N-methylmorpholine (2
24 mg, 2.21 mmol) was added, and the mixture was stirred at the same temperature for 1 hour and further at room temperature for 3 hours. The reaction mixture was concentrated under reduced pressure, ethyl acetate and water were added to the residue, and the insoluble matter (DC-Ur
ea) was filtered off. The organic layer was separated from the filtrate and washed successively with 10% aqueous citric acid solution, saturated aqueous sodium hydrogen carbonate and saturated brine,
It was dried over anhydrous sodium sulfate. After evaporating the solvent under reduced pressure, the residue was purified by medium pressure silica gel column chromatography (chloroform / methanol = 30/1) to obtain the title compound as a white powder (658 mg). ¹ H NMR (400 MHz, CDCl ₃ ) δ 1.28 (3 H, t, J = 7 Hz) 3.57 (1 H, d, J = 2 Hz) 3.63 (1 H, d, J = 2 Hz) 3.98 ( 1H, dd, J = 18,5Hz) 4.04 (1H, dd, J = 18,5Hz) 4.21 (1H, q, J = 7Hz) 6.22 (1H, d, J = 8Hz) 6. 54 (1H, br.t, J = 5Hz) 6.70 (1H, br.d, J = 8Hz) 7.1 to 7.4 (10H, m)

(3-6) Synthesis of N-[(2S, 3S) -3-diphenylmethylcarbamoyloxirane-2-carbonyl] glycine The compound (648 mg, 1.69 mmol) obtained in the above (3-5) and The title compound was obtained as a white powder by reacting and treating in the same manner as in (1-5) of Example 1 using a 0.5 N potassium hydroxide / ethanol solution (4.1 mL, 2.05 mmol) (586 mg) ). ¹ H NMR (400 MHz, CDCl ₃ -CD ₃ O
D) δ 3.22 (1H, d, J = 2Hz) 3.66 (1H, d, J = 2Hz) 3.92 (1H, d, J = 18Hz) 4.04 (1H, d, J = 18 Hz) 6.23 (1H, s) 7.2-7.4 (10H, m)

(3-7) [4- [N-[(2S, 3S)
-3-Diphenylmethylcarbamoyloxirane-2-
Synthesis of tetrabenzyl [carbonyl] glycyl] oxypiperidino] methylenebisphosphonate In a suspension of the compound (177 mg, 0.50 mmol) obtained in (3-6) above in anhydrous methylene chloride (10 mL),
Tetrabenzyl (4-hydroxypiperidino) methylenebisphosphonate obtained in (3-4) (318 mg, 0.5
0 mmol), 4-dimethylaminopyridine (12 m
g, 0.1 mmol) and DCC (144 mg, 0.55)
(Mmol), and the mixture was stirred at room temperature overnight. Insoluble matter (DC-Ur
ea) was filtered off, and the filtrate was washed successively with 10% aqueous citric acid solution, saturated aqueous sodium hydrogen carbonate solution and saturated brine, and dried over anhydrous sodium sulfate. After evaporating the solvent under reduced pressure, the residue was purified by medium pressure silica gel column chromatography (n-hexane / ethyl acetate = 1/10) to obtain the title compound as a pale brown viscous oil (267 mg). ¹ H NMR (400 MHz, CDCl ₃ ) δ 1.5 to 1.8 (4H, m) 2.8 to 3.0 (2H, m) 3.0 to 3.2 (2H, m) 3.40 ( 1H, t, J = 25Hz) 3.63 (1H, d, J = 2Hz) 3.70 (1H, d, J = 2Hz) 3.88 (1H, dd, J = 18,5Hz) 4.04 ( 1H, dd, J = 18,5Hz) 4.79 (1H, m) 4.9-5.1 (8H, m) 6.25 (1H, d, J = 8Hz) 6.53 (1H, br. t, J = 5 Hz) 7.00 (1 H, d, J = 8 Hz) 7.1-7.4 (30 H, m) FAB MS (m / z, relative intensity) 972 ([M + H] ⁺ , 5) 638 _{([M-PO 3 Bn 2} ] +, 35)

Example 4 [4- [N-[(2S, 3S) -3-diphenylmethylcarbamoyloxirane-2-carbonyl] glycyl]
Oxypiperidino] methylenebisphosphonic acid disodium salt Disodium salt of compound example (2) Compound obtained in Example 3 (3-7) (152 mg, 0.
156 mmol) was subjected to catalytic hydrogenation and sodium chloride in the same manner as in Example 2 to obtain the title compound as a light brown powder (101 mg). ¹ H NMR (400 MHz, D ₂ O) δ 2.0 to 2.3 (4H, m) 3.36 (1H, t, J = 17z) 3.6 to 3.8 (2H, m) 3.80 (1H, d, J = 2Hz) 3.85 (1H, d, J = 2Hz) 3.8-4.0 (2H, m) 4.15 (2H, s) 5.13 (1H, m) 6 .17 (1H, s) 7.3 to 7.5 (10H, m) FAB MS (m / z, relative intensity) 606 ([M + Na] ⁺ , 5) 584 ([M + H] ⁺ , 6) IR (KBr ) Cm ⁻¹ : 3408, 3032, 174
5,1670,1541,1535,1203,111
5,1036,895,700,544

Example 5 [4-[(2S, 3S) -3-Diphenylmethylcarbamoyloxirane-2-carbonyl] oxypiperidino] methylenebisphosphonic acid tetrabenzyl Compound Example (35) Obtained in Example 1 (1-5). (2S, 3S) -3-diphenylmethylcarbamoyloxirane-2-carboxylic acid (110 mg, 0.370 mmol), tetrabenzyl (4-hydroxypiperidino) methylenebisphosphonate (235 mg, 0.370 mmol), 4 -Dimethylaminopyridine (9 mg, 0.07 mmol) and DCC
By using (85 mg, 0.41 mmol) and reacting and treating in the same manner as in (3-7) of Example 3, the title compound was obtained as a white amorphous substance (262 mg). ¹ H NMR (400 MHz, CDCl ₃ ) δ 1.5 to 1.8 (4H, m) 2.8 to 3.0 (2H, m) 3.1 to 3.2 (2H, m) 3.38 ( 1H, t, J = 25Hz) 3.50 (1H, d, J = 2Hz) 3.86 (1H, d, J = 2Hz) 4.78 (1H, m) 4.9-5.1 (8H, m) 6.25 (1H, d, J = 8Hz) 7.00 (1H, d, J = 8Hz) 7.1-7.4 (30H, m). FAB MS (m / z, relative intensity) 915 ([M + H] ⁺ , 6) 653 ([M-PO ₃ Bn ₂ ] ⁺ , 30)

Example 6 [4-[(2S, 3S) -3-Diphenylmethylcarbamoyloxirane-2-carbonyl] oxypiperidino] methylenebisphosphonic acid disodium salt Disodium salt of compound example (3) Compound obtained in example 5 To a solution of (252 mg, 0.275 mmol) in ethyl acetate (4 mL) was added water (4 mL) and 1
0% Palladium / carbon (40 mg) was added, and catalytic hydrogenation was performed at room temperature and 1 atm. After 4 hours, sodium hydrogen carbonate (46.2 mg, 0.550 mmol) and water (2 mL) were added to the reaction mixture, the mixture was stirred for 15 minutes, and the catalyst was filtered off. The aqueous layer was separated, concentrated under reduced pressure to about 3 mL, and ethanol (about 10 mL) was added. The precipitate was collected by filtration, washed with ethanol, and dried under reduced pressure at 50 ° C to give the title compound as a white powder (132 mg). ¹ H NMR (400 MHz, D ₂ O) δ 2.0 to 2.3 (4 H, m) 3.40 (1 H, t, J = 17 Hz) 3.6 to 3.8 (2 H, m) 3.89 (1H, d, J = 2Hz) 3.93 (1H, d, J = 2Hz) 3.8-4.0 (2H, m) 5.19 (1H, m) 6.17 (1H, s) 7 .3 to 7.5 (10 H, m) FAB MS (m / z, relative intensity) 621 ([M + Na] ⁺ , 5) 599 ([M + H] ⁺ , 10) IR (KBr) cm ⁻¹ : 3408, 3032 174
3,1672,1533,1497,1454,128
4,1200,1095,1030,985,897,
700,540

Example 7 [4- [N-[(2S, 3S) -3-diphenylmethylcarbamoyloxirane-2-carbonyl] glycyl]
Tetrabenzyl [-1-piperazinyl] methylenebisphosphonate Compound Example (36) The compound obtained in Example 3 (3-6) (177 mg, 0.
50 mmol), N-hydroxysuccinimide (63
mg, 0.55 mmol), DCC (114 mg, 0.5
5 mmol), tetrabenzyl (1-piperazinyl) methylenebisphosphonate obtained in (1-3) of Example 1 p
A mixture of -toluenesulfonate and tribenzyl (1-piperazinyl) methylenebisphosphonate p-toluenesulfonate (382 mg, former compound; 262 m)
g, 0.33 mmol, latter compound; 120 mg,
0.17 mmol) and N-methylmorpholine (56 m
g, 0.55 mmol) of (1-6) of Example 1
Reaction and treatment were conducted in the same manner as in to give the title compound as a white amorphous substance (239 mg). ¹ H NMR (400 MHz, CDCl ₃ ) δ 2.92 (4 H, m) 3.12 (2 H, m) 3.34 (1 H, t, J = 25 Hz) 3.40 (2 H, m) 3.55 ( 3. 1 (H, d, J = 2Hz) 3.65 (1H, d, J = 2Hz) 3.90 (1H, dd, J = 17,4Hz) 3.92 (1H, dd, J = 17,4Hz) 9-5.1 (8H, m) 6.23 (1H, d, J = 8Hz) 6.71 (1H, d, J = 8Hz) 7.03 (1H, br.t, J = 4Hz) 7. 1-7.4 (30, m) FAB MS (m / z, relative intensity) 957 ([M + H] ⁺ , 2) 638 ([M-PO ₃ Bn ₂ ] ⁺ , 9)

Example 8 [4- [N-[(2S, 3S) -3-diphenylmethylcarbamoyloxirane-2-carbonyl] glycyl]
-1-Piperazinyl] methylenebisphosphonic acid disodium salt Disodium salt of compound example (4) The compound (229 mg, 0.240 mmol) obtained in Example 7 was subjected to catalytic hydrogenation and sodium chloride in the same manner as in Example 6. The title compound was obtained as a white powder (1
30 mg). ¹ H NMR (400 MHz, D ₂ O) δ 3.39 (1 H, t, J = 18 Hz) 3.6-4.0 (8 H, m) 3.80 (1 H, d, J = 2 Hz) 3.85 (1H, d, J = 2Hz) 4.26 (2H, s) 6.17 (1H, s) 7.3 to 7.5 (10H, m) FAB MS (m / z, relative intensity) 663 ([ M + Na] ⁺ , 3) 641 ([M + H] ⁺ , 7). IR (KBr) cm ^-1 : 3390,3062,166
4,1535,1495,1452,1240,117
4,1093,1061,895,700,544

Example 9 [4- [N-[(2S, 3S) -3-ethoxycarbonyloxirane-2-carbonyl] -L-leucyl] -1
-Piperazinyl] methylenebisphosphonate tetrabenzyl Compound example (37) [N-[(2S, 3S) -3-ethoxycarbonyloxirane-2-carbonyl] -L-leucine (151m
g, 0.55 mmol), HOSu (63 mg, 0.55 mmol), DCC (114 mg, 0.55 mmol), tetrabenzyl (1-piperazinyl) methylenebisphosphonate obtained in (1-3) of Example 1. Mixture of p-toluenesulfonate and tribenzyl (1-piperazinyl) methylenebisphosphonate p-toluenesulfonate (3
82 mg, former compound content; 262 mg, 0.33 mmol, latter compound content; 120 mg, 0.17 mmol) and N-methylmorpholine (56 mg, 0.55)
(1-6) of Example 1 to give the title compound as a colorless viscous oil (152 mg). ¹ H NMR (400 MHz, CDCl ₃ ) δ 0.88 (3H, d, J = 6 Hz) 0.95 (3H, d, J = 7 Hz) 1.2 to 1.6 (3H, m) 1.31 ( 3H, t, J = 7Hz) 2.8-3.0 (4H, m) 3.2-3.3 (2H, m) 3.3-3.5 (2H, m) 3.38 (1H, t, J = 25 Hz) 3.46 (1H, d, J = 2 Hz) 3.64 (1H, d, J = 2 Hz) 4.2-4.3 (2H, m) 4.86 (1H, m) 5.0-5.1 (8H, m) 7.2-7.4 (20H, m)

Example 10 [4- [N-[(2S, 3S) -3-ethoxycarbonyloxirane-2-carbonyl] -L-leucyl] -1
-Piperazinyl] methylenebisphosphonate disodium salt Disodium salt of compound example (12) The compound (142 mg, 0.162 mmol) obtained in Example 9 was subjected to catalytic hydrogenation and sodium chloride in the same manner as in Example 6 to give the title compound. Obtained as a white powder (75 mg). ¹ H NMR (400 MHz, D ₂ O) δ 0.94 (3H, d, J = 6 Hz) 0.95 (3H, d, J = 6 Hz) 1.53 (1H, m) 1.67 (2H, m ) 3.41 (1H, t, J = 18Hz) 3.6 to 4.0 (8H, m) 3.76 (1H, d, J = 2Hz) 3.82 (1H, d, J = 2Hz) 4 .31 (2H, q, J = 7 Hz) 4.91 (1H, dd, J = 10, 4 Hz) FAB MS (m / z, relative intensity) 582 ([M + Na] ⁺ , 12) 560 ([M + H] ⁺ , 18) IR (KBr) cm ⁻¹ : 3398, 3390, 296
0,2872,1745,1645,1541,153
5,1471,1448,1371,1308,127
7,1205,1095,1020,958,899,
542

Example 11 [4- [N-[(2S, 3S) -3-Carboxyoxirane-2-carbonyl] -L-leucyl] -1-piperazinyl] methylenebisphosphonic acid tetrabenzyl Compound Example (31) Example 9 (187 mg, 0.213 mmol) and a 0.5 N potassium hydroxide / ethanol solution (0.
The reaction and treatment were carried out in the same manner as in (1-5) of Example 1 using 47 mL (0.235 mmol) to give the title compound as a colorless candy (117 mg). ¹ H NMR (400 MHz, CDCl ₃ ) δ 0.91 (3H, d, J = 6 Hz) 0.93 (3H, d, J = 6 Hz) 1.37 (1H, m) 1.5 to 1.7 ( 2H, m) 2.8-3.1 (4H, m) 3.2-3.3 (2H, m) 3.34 (1H, t, J = 25Hz) 3.5-3.6 (2H, m) m) 3.56 (1H, d, J = 2Hz) 3.65 (1H, d, J = 2Hz) 4.9 to 5.1 (9H, m) 7.2 to 7.4 (20H) 8. 13 (1H, d, J = 9Hz)

Example 12 [4- [N-[(2S, 3S) -3-Carboxyoxirane-2-carbonyl] -L-leucyl] -1-piperazinyl] methylenebisphosphonate trisodium Compound Example (15) Sodium salt The compound (172 mg, 0.203 mmol) obtained in Example 11 was subjected to catalytic hydrogenation and sodium chloride in the same manner as in Example 6 to obtain the title compound as a white powder (108 m
g). ¹ H NMR (400 MHz, D ₂ O) δ 0.94 (3H, d, J = 6 Hz) 0.95 (3H, d, J = 6 Hz) 1.52 (1H, m) 1.6 to 1.8 (2H, m) 3.41 (1H, t, J = 18Hz) 3.45 (1H, d, J = 2Hz) 3.59 (1H, d, J = 2Hz) 3.6-6.0 (8H) , M) 4.90 (1 H, dd, J = 10, 4 Hz). FAB MS (m / z, relative intensity) 576 ([M + Na] ⁺ , 6) 554 ([M + H] ⁺ , 6) IR (KBr) cm ⁻¹ : 3421, 2960, 287
1,1632, 1554, 1444, 1387, 124
2,1155,1095,958,899,544,4
63

Example 13 [4- [N-[(2S, 3S) -3-ethoxycarbonyloxirane-2-carbonyl] -L-leucyl-L-
Leucyl] -1-piperazinyl] methylenebisphosphonate tetrabenzyl Compound example (38) N-[(2S, 3S) -3-ethoxycarbonyloxirane-2-carbonyl] -L-leucyl-L-leucine (387 mg, 1.00) Millimoles), HOSu (116m
g, 1.0 mmol), DCC (207 mg, 1.0 mmol), tetrabenzyl p-toluenesulfonate (1-piperazinyl) methylenebisphosphonate obtained in (1-3) of Example 1 and (1- Piperazinyl) methylenebisphosphonate tribenzyl p-toluenesulfonate mixture (767 mg, former compound content; 547 mg,
0.69 mmol, content of the latter compound; 220 mg,
0.31 mimolole) and N-methylmorpholine (113
mg, 1.11 mmol), and
Reaction and treatment were conducted in the same manner as in 6) to give the title compound as a colorless viscous oil (556 mg). ¹ H NMR (400 MHz, CDCl ₃ ) δ 0.88 (3H, d, J = 6 Hz) 0.92 (6H, d, J = 6 Hz) 0.95 (3H, d, J = 6 Hz) 1.31 ( 3H, t, J = 7 Hz) 1.3 to 1.6 (6H, m) 2.9 to 3.1 (4H, m) 3.2 to 3.5 (4H, m) 3.38 (1H, t, J = 25 Hz) 3.45 (1 H, d, J = 2 Hz) 3.68 (1 H, d, J = 2 Hz) 4.2-4.3 (2 H, m) 4.45 (1 H, m) 4.83 (1H, m) 5.0 to 5.1 (8H, m) 6.54 (1H, d, J = 8Hz) 6.62 (1H, d, J = 9Hz) 7.2 to 7. 4 (20H, m) FAB MS (m / z, relative intensity) 989 ([M + H] ⁺ , 4) 727 ([M-PO ₃ Bn ₂ ] ⁺ , 37)

Example 14 [4- [N-[(2S, 3S) -3-Ethoxycarbonyloxirane-2-carbonyl] -L-leucyl-L-
[Leucyl] -1-piperazinyl] methylenebisphosphonate disodium salt Disodium salt of compound example (16) The compound (289 mg, 0.292 mmol) obtained in Example 13 was subjected to catalytic hydrogenation and sodium chloride in the same manner as in Example 2. To give the title compound as a white powder (196 m
g). ¹ H NMR (400 MHz, D ₂ O) δ 0.8 to 1.0 (12 H, m) 1.31 (3 H, t, J = 7 Hz) 1.4 to 1.7 (6 H, m) 3.43 (1H, t, J = 18 Hz) 3.74 (1H, s) 3.80 (1H, s) 3.6-4.0 (8H, m) 4.31 (1H, q, J = 7Hz) 4 .44 (1H, m) 4.87 (1H, dd, J = 10, 4 Hz) FAB MS (m / z, relative intensity) 695 ([M + Na] ⁺ , 15) 673 ([M + H] ⁺ , 19) IR (KBr) cm ⁻¹ : 3398, 2958, 287
2,1743,1655,1647,1541,153
5,1470,1448,1369,1277,120
5,1163,1095,1030,899,544

Example 15 [4- [N-[(2S, 3S) -3-Carboxyoxirane-2-carbonyl] -L-leucyl-L-leucyl] -1-piperazinyl] methylenebisphosphonate tetrabenzyl Compound Example ( 39) Using the compound obtained in Example 13 (214 mg, 0.216 mmol) and 0.5N potassium hydroxide / ethanol solution (0.48 mL, 0.24 mmol), Example 1
The reaction and treatment were carried out in the same manner as in (1-5) of (1) to give the title compound as a white amorphous substance (202 mg). ¹ H NMR (400 MHz, CDCl ₃ ) δ 0.8-1.0 (12H, m) 1.3-1.7 (6H, m) 2.8-3.1 (4H, m) 3.2 3.5 (4H, m) 3.37 (1H, t, J = 25Hz) 3.56 (1H, d, J = 1Hz) 3.62 (1H, d, J = 1Hz) 4.54 (1H, m) 4.89 (1H, m) 5.0 to 5.12 (8H, m) 7.05 (1H, br) 7.2 to 7.4 (21H, m)

Example 16 [4- [N-[(2S, 3S) -3-Carboxyoxirane-2-carbonyl] -L-leucyl-L-leucyl] -1-piperazinyl] methylenebisphosphonate trisodium Compound Example ( Trisodium salt of 17) The compound (195 mg, 0.203 mmol) obtained in Example 15 was subjected to catalytic hydrogenation and sodium chloride in the same manner as in Example 6 to obtain the title compound as a white powder (108 m).
g). ¹ H NMR (400 MHz, D ₂ O) δ 0.8 to 1.0 (12 H, m) 1.4 to 1.8 (6 H, m) 3.41 (1 H, t, J = 18 Hz) 3.44 (1H, d, J = 2Hz) 3.57 (1H, d, J = 2Hz) 3.6-4.0 (8H, m) 4.42 (1H, m) 4.88 (1H, dd, J) = 10,4 Hz) FAB MS (m / z, relative intensity) 689 ([M + Na] ⁺ , 4) 667 ([M + H] ⁺ , 5) IR (KBr) cm ^-1 : 3427,2958,287
1,1631, 1541, 1535, 1468, 144
8,1387,1246,1155,1115,109
7,899,544,463

Example 17 [3- [4- [N-[(2S, 3S) -3-ethoxycarbonyloxirane-2-carbonyl] -L-leucyl]-
1-Piperazinyl] -3-oxo] propane-1,1-
Tetrabenzyl Bisphosphonate Compound Example (40) (17-1) 2- (t-butoxycarbonyl) ethyl-
Synthesis of tetrabenzyl 1,1-bisphosphonate Under a nitrogen atmosphere, tetrabenzyl methylenebisphosphonate (3.45 g, 6.43 mmol) was dissolved in dry tetrahydrofuran (17 mL), and 60% sodium hydride (514 mg, 12.9 mmol) was added and the mixture was stirred at the same temperature for 20 minutes. To this, t-butyl bromoacetate (0.95 mL, 6.43 mol) was added under ice cooling, and the mixture was stirred at room temperature for 1 hour and 40 minutes. The reaction solution was poured into saturated aqueous ammonium chloride solution (50 mL) and extracted with chloroform (50 mL × 3). The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. After evaporating the solvent under reduced pressure, the residue was purified by medium pressure silica gel column chromatography (hexane / ethyl acetate = 1/1 to 1/2) to give the title compound as a colorless oil (2.80).
g). ¹ H NMR (400 MHz, CDCl ₃ ) δ 1.34 (9 H, s) 2.83 (2 H, dt, J = 6.16 Hz) 3.31 (2 H, tt, J = 6.24 Hz) 5.0- 5.1 (8H, m) 7.2-7.4 (20H, m)

(17-2) 2-carboxyethyl-1,
Synthesis of tetrabenzyl 1-bisphosphonate The compound (2.80 g, 4.30) obtained in the above (17-1).
Mmol) was dissolved in 98% formic acid (10 mL) and stirred at room temperature for 3 hours. Formic acid was distilled off under reduced pressure and then cooled to 10%
A saturated aqueous sodium hydrogen carbonate solution (50 mL) was added, and the mixture was extracted with ethyl acetate (50 mL × 2). The ethyl acetate layer was dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure to give the title compound as a colorless oil (2.12 g). ¹ H NMR (400 MHz, CDCl ₃ ) δ 2.91 (2H, dt, J = 6.16 Hz) 3.29 (1H, tt, J = 6.24 Hz) 4.9-5.0 (8H, m) 7.2-7.3 (20H, m)

(17-3) (2S, 3S) -3-
Synthesis of ethyl [[[(S) -3-methyl-1- [4- (tert-butoxycarbonyl) -1-piperazinyl] carbonyl] butyl] carbamoyl] oxirane-2-carboxylate N-[(2S, 3S) -Ethoxycarbonyloxirane-2-carbonyl] -L-leucine (2.23 g, 7.
32 mmol), tert-butyl 1-piperazinecarboxylate (1.50 g, 8.05 mmol), 1-hydroxybenzotriazole (1.09 g, 8.05 mmol), N-methylmorpholine (0.88 mL, 8. 00
Mmol) was dissolved in dry tetrahydrofuran (50 mL), and 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (1.54 g, 8.0) under ice cooling.
5 mmol) was added slowly. After stirring at the same temperature for 2 hours, the mixture was stirred at room temperature for 3 hours. The solvent was evaporated under reduced pressure, water (100 mL) and ethyl acetate (100 mL) were added, and the organic layer was separated. The aqueous layer was washed with ethyl acetate (50 mL x
The mixture was extracted with 2), combined with the above organic layer, washed successively with a 10% aqueous citric acid solution, a saturated aqueous sodium hydrogen carbonate solution and a saturated saline solution, and then dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure, and the residue was purified by medium pressure silica gel column chromatography (chloroform / methanol = 50/1) to give the title compound as a white amorphous substance (2.02 g). ¹ H NMR (400 MHz, CDCl ₃ ) δ 0.91 (3H, d, J = 7 Hz) 0.98 (3H, d, J = 7 Hz) 1.32 (3H, t, J = 7 Hz) 1.4- 1.6 (3H, m) 1.47 (9H, s) 3.4 to 3.7 (8H, m) 3.47 (1H, d, J = 2Hz) 3.66 (1H, d, J = 2Hz) 4.2-4.3 (2H, m) 4.95 (1H, m) 6.83 (1H, br.d, J = 9Hz)

(17-4) (2S, 3S) -3-
[[[(S) -3-Methyl-1- (1-piperazinyl))
Carbonyl] butyl] carbamoyl] oxirane-2-
Synthesis of ethyl carboxylate (2S, 3S) -3-obtained in (17-3) above.
Ethyl [[[(S) -3-methyl-1-[[4- (tert-butoxycarbonyl) -1-piperazinyl] carbonyl] butyl] carbamoyl] oxirane-2-carboxylate (2.02 g, 4.58 mmol) Was dissolved in trifluoroacetic acid (4 mL), and the mixture was stirred at 0 ° C. for 1 hr 30 min. After the solvent was distilled off under reduced pressure, the residue was chloroform (60 m
L) and saturated aqueous sodium hydrogen carbonate solution (50
(mL) and saturated brine successively. After drying over anhydrous sodium sulfate, the solvent was evaporated under reduced pressure to give the title compound as a white amorphous substance (1.51 g). ¹ H NMR (400 MHz, CDCl ₃ ) δ 0.91 (3H, d, J = 7 Hz) 0.98 (3H, d, J = 7 Hz) 1.31 (3H, t, J = 7 Hz) 1.4- 1.7 (3H, m) 2.8 to 3.0, 3.4 to 3.6 (8H, m) 3.47 (1H, d, J = 2Hz) 3.66 (1H, d, J = 2Hz) 4.2-4.4 (2H, m) 4.97 (1H, m) 6.93 (1H, br.d, J = 9Hz)

(17-5) [3- [4- [N-[(2
S, 3S) -3-Ethoxycarbonyloxirane-2-
Carbonyl] -L-leucyl] -1-piperazinyl]-
Synthesis of tetrabenzyl 3-oxo] propane-1,1-bisphosphonate Tetrabenzyl 2-carboxyethyl-1,1-bisphosphonate obtained in (17-2) (595 mg, 1.00 mmol) and HOSu (115 mg, 1.00 mmol) was dissolved in ethyl acetate (2 mL) and DCC (206 mL) was added under ice cooling.
mg, 1.00 mmol) in ethyl acetate (1 mL) was added, and the mixture was stirred at room temperature for 30 minutes. In addition to this, (2S, 3S) -3-[[[[(S) -3-methyl-1-
Ethyl (1-piperazinyl) carbonyl] butyl] carbamoyl] oxirane-2-carboxylate (431 mg,
A solution of 1.00 mmol) in ethyl acetate (1 mL) was added under ice-cooling, and the mixture was stirred at 5 ° C. for 1 hr and then at room temperature overnight. The deposited insoluble material (DC-Urea, N, N'-dicyclohexylurea) was filtered off and washed with ethyl acetate. The filtrate and the washing solution were combined, washed successively with a 10% aqueous citric acid solution, a saturated aqueous sodium hydrogen carbonate solution and a saturated saline solution, and then dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure, and the residue was purified by medium pressure silica gel column chromatography (ethyl acetate / methanol = 50/1) to give the title compound as a white amorphous substance (164 mg). ¹ H NMR (400 MHz, CDCl ₃ ) δ 0.91 (3H, d, J = 7 Hz) 0.97 (3H, d, J = 7 Hz) 1.32 (3H, t, J = 7 Hz) 1.4- 1.7 (3H, m) 2.7 to 2.9 (2H, m) 3.1 to 3.6 (8H, m) 3.46 (1H, d, J = 2Hz) 3.66 (1H, d, J = 2Hz) 3.69 (1H, tt, J = 6,24Hz) 4.2-4.3 (2H, m) 4.87 (1H, m) 4.9-5.1 (8H, m) 6.79 (1H, br.d, J = 9Hz) 7.2-7.4 (20H, m)

Example 18 [3- [4- [N-[(2S, 3S) -3-Carboxyoxirane-2-carbonyl] -L-leucyl] -1-
Piperazinyl] -3-oxo] propane-1,1-bisphosphonate tetrabenzyl Compound Example (41) The compound obtained in Example 17 (17-5) (164 mg,
0.178 mmol) and 0.5 N potassium hydroxide / ethanol solution (0.39 mL, 0.195 mmol) were used to react and treat in the same manner as in (1-5) of Example 1,
The title compound was obtained as a white amorphous (150 m
g). ¹ H NMR (400 MHz, CDCl ₃ ) δ 0.93 (3H, d, J = 7 Hz) 0.96 (3H, d, J = 7 Hz) 1.5 to 1.8 (3H, m) 2.80 ( 2H, dt, J = 6, 16Hz) 3.2-3.5 (6H, m) 3.5-3.6 (4H, m) 3.67 (1H, tt, J = 6, 24Hz) 4. 9-5.1 (9H, m) 7.2-7.4 (20, m) 7.69 (1H, br)

Example 19 [3- [4- [N-[(2S, 3S) -3-Carboxyoxirane-2-carbonyl] -L-leucyl] -1-
Piperazinyl] -3-oxo] propane-1,1-bisphosphonic acid trisodium salt Sodium salt of compound example (18) The compound (150 mg, 0.169 mmol) obtained in Example 18 was contacted with hydrogen in the same manner as in Example 6. Addition and sodium chloride gave the title compound as a white powder (97.0m
g). ¹ H NMR (400 MHz, D ₂ O) δ 0.94 (6 H, d, J = 6 Hz) 1.5 to 1.8 (3 H, m) 2.68 (1 H, tt, J = 6, 24 Hz) 2 .8 to 3.0 (2H, m) 3.45 (1H, d, J = 2Hz) 3.59 (1H, d, J = 2Hz) 3.5 to 3.9 (8H, m) 4.89 (1H, m) IR (KBr) cm ^-1 : 3415,3275,295
8,2872,1628,1470,1443,128
7,1282,1246,1169,1086,101
6,897,766,530,467

Example 20 [4-[(2S, 3S) -3-[(S) -3-methyl-]
1- (3-Methylbutylcarbamoyl) butylcarbamoyl] oxirane-2-carbonyl] oxypiperidino] methylenebisphosphonate tetrabenzyl Compound example (42) (2S, 3S) -3-[(S) -3-methyl-1- ( Three
-Methylbutylcarbamoyl) butylcarbamoyl]-
2-oxiranecarboxylic acid (190 mg, 0.605 mmol), tetrabenzyl (4-hydroxypiperidino) methylenebisphosphonate (350 mg, 0.550 mmol), 4-dimethylaminopyridine (14.7 mg,
0.120 mmol) and DCC (125 mg, 0.60
5 mmol) was used and reacted and treated in the same manner as in (3-7) of Example 3 to obtain the title compound as a white amorphous substance (457 mg). ¹ H NMR (400 MHz, CDCl ₃ ) δ 0.9 to 1.0 (12 H, m) 1.39 (2 H, q, J = 7 Hz) 1.5 to 1.9 (6 H, m) 2.9 to 3.0 (2H, m) 3.1 to 3.3 (4H, m) 3.40 (1H, t, J = 24Hz) 3.45 (1H, d, J = 2Hz) 3.78 (1H, d, J = 2 Hz) 4.38 (1H, m) 4.78 (1H, m) 5.0 to 5.1 (8H, m) 6.02 (1H, br) 6.91 (1H, br. d, J = 8 Hz) 7.2-7.4 (20H, m)

Example 21 [4-[(2S, 3S) -3-[(S) -3-Methyl-]
1- (3-Methylbutylcarbamoyl) butylcarbamoyl] oxirane-2-carbonyl] oxypiperidino] methylenebisphosphonate disodium salt Disodium salt of compound example (6) The compound obtained in example 20 (457 mg, 0.490 mmol) was used. Catalytic hydrogenation and sodium chloride as in Example 6 gave the title compound as a white powder (275mg). ¹ H NMR (400 MHz, D ₂ O) δ 0.8 to 1.0 (12 H, m) 1.3 to 1.7 (6 H, m) 2.1 to 2.3 (4 H, m) 3.1 -3.3 (2H, m) 3.41 (1H, t, J = 17Hz) 3.6-4.0 (4H, m) 3.82 (1H, br.s) 3.87 (1H, br) .S) 4.32 (1H, m) 5.19 (1H, m) IR (KBr) cm ⁻¹ : 3398, 3304, 308
8, 2958, 2872, 1745, 1653, 155
1, 1470, 1387, 1350, 1282, 119
8,1095,1028,987,897,540

Example 22 Bone resorption inhibitory effect [0137] Calvariae were aseptically collected from 6-7 day-old ICR mice to remove connective tissue, and then the calvariae were cut in half along the midline. 1 mL of culture medium per pair of bones (modif
ied BGJb medium, containing 5% inactivated fetal bovine serum)
24 hours preculture (5% CO ₂ , 37 ° C.) was performed. After the pre-culture, each piece of bone was placed in 0.5 mL of the above culture solution containing various concentrations of the test compound, and cultured for 72 hours in the presence of parathyroid hormone (PTH) 3 × 10 ⁻⁷ M. In addition, the cells cultured in the absence of the test compound and PTH served as controls. The amount of calcium released in the culture supernatant and the amount of calcium in the bone (dissolved in 1 mL of 6N hydrochloric acid) during 72 hours after the completion of the culture were measured using the orthocresolphthalein complexone (OCPC) method. First, the calcium release rate (%) was calculated according to the following formula.

[0138]

[Equation 1]

Next, the bone resorption inhibition rate (%) by the test compound was determined according to the following formula.

[0140]

[Equation 2]

The above results are shown in Table 1 below.

[0142]

[Table 1] Table 1 ──────────────────────────────────── Test compound bone resorption inhibitory effect (Test compound concentration) 10 ⁻⁷ M 10 ⁻⁶ M 10 ⁻⁵ M 10 ⁻⁴ M ──────────────────────────── ──────── Disodium salt of (1) 28 20 77 120 Disodium salt of (2) 26 56 109 Disodium salt of (3) 30 57 89 Disodium salt of (4) 40 60 98 ( 12) disodium salt 54 85 (15) trisodium salt 57 77 91 (16) disodium salt 42 60 73 (17) trisodium salt 38 53 79 (18) trisodium salt 28 51 72 ───────────────────────────────────

Example 23 Cathepsin L inhibitory action (23-1) Preparation of rat liver lysosomal fraction Male Wistar rats were killed by exsanguination, and ice-cooled physiological saline was injected from the portal vein for perfusion, and the liver was then removed. I removed it. The following operation was performed at 4 ° C. After finely cutting with scissors, 5 g was weighed, and 9 times amount of 0.25 M sucrose solution was added to homogenize (Potter type Teflon homogenizer).
The supernatant obtained by centrifuging the homogenate at 800 xg for 10 minutes was further centrifuged at 12000 xg for 20 minutes. 25 mL of a 0.25 M sucrose solution was added to the obtained precipitate and the mixture was homogenized and then centrifuged at 12000 × g for 20 minutes. The resulting precipitate was homogenized by adding 10 ml of 0.25M sucrose solution to obtain a lysosomal fraction. This fraction was diluted with a 0.25 M sucrose solution containing 0.33% Triton X-100 and used for the measurement of cathepsin L activity. (23-2) Measurement of cathepsin L activity 340 mM sodium acetate, 60 mM acetic acid, 4 mM EDTA
And a solution containing 8 mM dithiothreitol (pH: 5.
5) To 0.25 mL, lysosomal fraction 0.1 mL, test compound solution 5 μL and distilled water 0.545 mL were added and pre-incubated at 30 ° C. for 15 minutes, and then 50 μM carbobenzoxy-L-phenylalanyl was used as a substrate. −
L-arginine-4-methylcoumaryl-7-amide (ZP
he-Arg-MCA) solution (0.1 mL) was added to start the reaction.
After reacting for 20 minutes at 30 ° C, 100 mM sodium monochloroacetate, 30 mM sodium acetate and 70 mM
The reaction was stopped by adding 1 mL of a solution containing acetic acid (pH: 4.3). The fluorescence intensity of the final solution is set to the excitation wavelength of 380 nm,
The fluorescence wavelength was measured at 460 nm. Z-Phe-Arg-MCA
Is also degraded by cathepsin B contained in the lysosomal fraction, and therefore CA is a cathepsin B-specific inhibitor.
-074 [Murata et al., FEBS Lett. 280 , 307-310 (1991).
] Was added to the reaction solution at 10 ⁻⁷ M to completely inhibit cathepsin B, and the measurement was performed.

The results are shown in Table 2 below.

[0145]

[Table 2] Table 2 ──────────────────────────────────── Test compound Cathepsin L inhibition IC ₅₀ (M) ──────────────────────────────────── (12) Disodium salt 2.4 ^{X10 -6} (15) trisodium salt 1.5 x 10 ^-7 (16) disodium salt 2.5 x 10 ^-8 (17) trisodium salt 5.4 x 10 ^-10 (18) Trisodium salt 1.9 × 10 ^-8 ─────────────────────────────────────

Example 24 Toxicity test 150 to 200 mg of Wistar rats were orally administered (100 mg / kg) with the compound of the present invention (the compound described in Example 6) for 2 weeks every day, but serious side effects were observed. Was not done.

Claims (15)

[Claims] 1. A bisphosphonic acid derivative having a partial structure of epoxysuccinic acid and exhibiting cysteine protease inhibitory activity. 2. A method having a partial structure of epoxysuccinic acid and a partial structure of a linking group, which releases an epoxysuccinic acid derivative showing cysteine protease inhibitory activity by being subjected to physicochemical conversion or metabolism in vivo. The bisphosphonic acid derivative according to item 1. 3. The carbonyl group of epoxysuccinic acid is bonded to the bisphosphonic acid structure by a linking group.
The bisphosphonic acid derivative according to the item. 4. A bisphosphonic acid derivative represented by the following formula (I) or a salt thereof: In the above formula, R ¹ is a hydrogen atom, an alkyl group or an aralkyl group; X ¹ is —O—, —NR ⁶ — or a divalent heterocyclic residue; A ¹ is a single bond or a right side. Is an N-terminal amino acid residue or dipeptide residue; A ² is a single bond or a left N-terminal amino acid residue or dipeptide residue; L ¹ is a divalent linking group; , R ² , R ³ , R ⁴ , R ⁵ and R
⁶ is each independently a hydrogen atom, an alkyl group or an aralkyl group. 5. In the formula (I), L ¹ is -O-,-
NR ⁷ — (R ⁷ is a hydrogen atom, an alkyl group, or an aralkyl group), a divalent heterocyclic residue, an alkylene group,
A divalent linking group selected from the group consisting of —CO—, a phenylene group, and combinations thereof.
A bisphosphonic acid derivative or a salt thereof according to the item 1. 6. A bisphosphonic acid derivative represented by the following formula (II) or a salt thereof: In the above formula, R ²¹ is an alkyl group or an aralkyl group; X ²¹ is —NR ²⁶ — or a divalent heterocyclic residue; A ²¹ is a single bond or an amino acid having the N-terminal on the right side. A ²² is a residue or a dipeptide residue; A ²² is a single bond or an amino acid residue having the N terminus on the left side;
L ²¹ is a divalent linking group selected from the group consisting of —O—, —NR ²⁷ —, a divalent heterocyclic residue, an alkylene group, —CO—, a phenylene group and combinations thereof; and R ²² , R ²³ , R ²⁴ , R ²⁵ , R ²⁶ and R ²⁷ are each independently a hydrogen atom, an alkyl group or an aralkyl group. 7. A bisphosphonic acid derivative represented by the following formula (III) or a salt thereof: In the above formula, R ³¹ is a hydrogen atom, an alkyl group or an aralkyl group; X ³¹ is —O— or —NR ³⁶ —.
A ³² is a single bond or an amino acid residue or dipeptide residue having the N-terminal on the left side; L ³¹ is —O
-, - NR ³⁷ -, a divalent heterocyclic residue, an alkylene group, -
A divalent linking group selected from the group consisting of CO--, a phenylene group, and combinations thereof; and R ³² ,
R ³³ , R ³⁴ , R ³⁵ , R ³⁶ and R ³⁷ are each independently a hydrogen atom, an alkyl group or an aralkyl group. 8. The amino acid residue or the dipeptide residue comprises a residue of glycine, alanine, valine, leucine, isoleucine, phenylalanine or tyrosine, or a combination thereof.
A bisphosphonic acid derivative or a salt thereof according to any one of items. 9. A bisphosphonic acid derivative or a salt thereof according to claim 4, wherein L ¹ is selected from the group consisting of the following L ^{2 to} L ¹⁵ : L ² : -heterocyclic residue-L ³ : - heterocyclic residue - alkylene - L ^4: -O- heterocyclic residue - L ^5: -O- alkylene ^{-CO-NR 8 - L 6:} -NR 9 - alkylene ^{-CO-NR 10 - L 7:} - O- alkylene - L ^8: - heterocyclic residue -CO- alkylene - L ^9: - heterocyclic residue -CO-O- alkylene - L ^10: - heterocyclic residue -O-CO- alkylene - L ^11: -NR ¹¹ - alkylene -NR ¹² -CO- alkylene - L ^12: - heterocyclic residue - alkylene - phenylene -O- alkylene - L ^13: - heterocyclic residue - alkylene - phenylene -O-C
O- alkylene - L ^14: - heterocyclic residue - alkylene - phenylene -O- alkylene ^{-CO-NR 13 - L 15:} -NR 14 - alkylene -O-CO- alkylene - In the above formulas, R ^8, R ⁹ , R ¹⁰ , R ¹¹ , R ¹² , R
¹³ and R ¹⁴ are each independently a hydrogen atom, an alkyl group or an aralkyl group. 10. The bisphosphonic acid derivative or a salt thereof according to claim 6, wherein L ²¹ is selected from the group consisting of the following L ^{22 to} L ²⁶ : L ²² : -heterocyclic residue-L ²³ : -O- heterocyclic residue - L ^24: -O- alkylene ^{-CO-NR 28 - L 25:} -NR 29 - alkylene ^{-CO-NR 30 - L 26:} -O- alkylene - in the above formulas, R ²⁸ , R ²⁹ and R ³⁰ are each independently a hydrogen atom, an alkyl group or an aralkyl group. 11. A bisphosphonic acid derivative or a salt thereof according to claim 7, wherein L ³¹ is selected from the group consisting of the following L ^{32 to} L ⁴¹ : L ³² : -heterocyclic residue-L ³³ : - heterocyclic residue - alkylene - L ^34: - heterocyclic residue -CO- alkylene - L ^35: - heterocyclic residue -CO-O- alkylene - L ^36: - heterocyclic residue -O-CO- alkylene - L ^37: -NR ³⁸ - alkylene -NR ³⁹ -CO- alkylene - L ^38: - heterocyclic residue - alkylene - phenylene -O- alkylene - L ^39: - heterocyclic residue - alkylene - phenylene -O-C
O- alkylene - L ^40: - heterocyclic residue - alkylene - phenylene -O- alkylene ^{-CO-NR 40 - L 41:} -NR 41 - alkylene -O-CO- alkylene - In the above formulas, R ^38, R ³⁹ , R ⁴⁰ and R ⁴¹ are each a hydrogen atom, an alkyl group or an aralkyl group. 12. A divalent heterocyclic residue having a piperidine ring or a piperazine ring, as defined in any one of claims 4 to 11.
A bisphosphonic acid derivative or a salt thereof according to any one of paragraphs. 13. Having a partial structure of epoxysuccinic acid,
A therapeutic agent for a bone disease, which comprises as an active ingredient a bisphosphonic acid derivative having a cysteine protease inhibitory activity or a physiologically acceptable salt thereof. 14. A therapeutic agent for a bone disease containing a bisphosphonic acid derivative represented by the following formula (I) or a physiologically acceptable salt thereof as an active ingredient: In the above formula, R ¹ is a hydrogen atom, an alkyl group or an aralkyl group; X ¹ is —O—, —NR ⁶ — or a divalent heterocyclic residue; A ¹ is a single bond or a right side. Is an N-terminal amino acid residue or dipeptide residue; A ² is a single bond or a left N-terminal amino acid residue or dipeptide residue; L ¹ is a divalent linking group; , R ² , R ³ , R ⁴ , R ⁵ and R
⁶ is each independently a hydrogen atom, an alkyl group or an aralkyl group. 15. In the formula (I), L ¹ is —O—,
—NR ⁷ — (wherein R ⁷ is a hydrogen atom, an alkyl group, or an aralkyl group), a divalent heterocyclic residue, an alkylene group, —CO—, a phenylene group, or a combination thereof. A therapeutic agent for bone diseases, which comprises a bisphosphonic acid derivative or a physiologically acceptable salt thereof according to claim 14 which is a divalent linking group as defined above.