JPH02104593A - Combination of medical compound with diphosphonic acid derivative - Google Patents

Abstract

NEW MATERIAL:A compound expressed by formula I (A-CO-represents residue of a medical compound; R represents -NH- or -O-; m is 0 or 1; n is 1-10) and salts thereof. EXAMPLE:Disodium salt of [alpha-(3',6'-dihydroxy-3-oxo-spiro[phthalan-1,9'- xanthene]-6-carbonyloxy)acetamidomethylene]bis(phosphonic acid). USE:An anti-inflammatory agent, remedy for osteopathy and carcinostatic agent. PREPARATION:A compound expressed by the formula A-COOH is reacted with a compound expressed by formula II (Y represents halogen) in an inert solvent such as water or methanol preferably in the presence of a base (example; sodium hydroxide or trimethylamine) at ambient temperature of while being heated to form a compound expressed by formula III, followed by elimination of a hydroxy-protecting group.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Field of Application] This invention relates to a conjugate of a pharmaceutical compound and a diphosphonic acid derivative (
(hereinafter referred to as 'conjugate'), more specifically, it relates to a conjugate in which a pharmaceutical compound having a carboxy group in the molecule or a salt thereof and a diphosphonic acid derivative are bound via a suitable spacer, and is used in the medical field. used.

[Prior art and its problems] Conventionally, when administering anti-inflammatory drugs to treat bone diseases such as rheumatoid arthritis, in order to deliver high-concentration anti-inflammatory drugs to the affected bone area, injections have been used to administer the drug into the joint. Direct administration is being practiced. However, since anti-inflammatory agents injected into a joint are quickly excreted from the injection site, the anti-inflammatory effect cannot be maintained for a long period of time.

In order to maintain the drug concentration at the administration site after intra-articular injection, attempts have been made to make the drug poorly water soluble [Clinical Pharmafukinetics (C11nica
l Pharmacokinetics), 8,
496-522, 1983, but nothing that fully achieves that purpose has yet been developed.

It has also been pointed out that in order to prevent cancer from metastasizing to bone, it is necessary to deliver anticancer drugs to bone tissue at high concentrations, but no drug has yet been developed that fully achieves this goal.

On the other hand, it has been reported that a complex of a diphosphonic acid derivative and technetium is synthesized and used for the diagnosis of arthritis [Journal of Nuclear Medicine].
cine), Volume 18 No. 10.973-976
page.

1977].

Furthermore, the concept of using hydroxyapatite [molecular formula: cat(po4)s(oH)z], which is a main component of bone, as a drug carrier has already been reported.

[Calcified Tissue International (C
a1cified Ti5sua International
onal), 40 volumes.

pp. 344-348, 1987].

[Means for Solving the Problems] The inventor of the present invention has conducted intensive research in order to obtain a derivative of a pharmaceutical compound that can maintain the concentration of the pharmaceutical compound in the bone for a long period of time. As a result, when a conjugate of a pharmaceutical compound or its salt having a carboxy group in its molecule and a diphosphonic acid derivative synthesized via various spacers is administered to a living body, it is selectively incorporated into bone tissue. It was found that the bone concentration was maintained at a high level for a long period of time.

Furthermore, this conjugate breaks the bond with the diphosphonic acid derivative in the bone tissue and is gradually converted into the above-mentioned pharmaceutical compound, allowing its concentration in the bone to be maintained at a high level for a long period of time. This invention was completed after discovering that the blood concentration could be maintained for a long period of time.

The conjugates of this invention are novel and have the form - where A-C
o- is a residue of a pharmaceutical compound, R is one book or -0-1m
is 0 or 1, and n is an integer from 1 to 10, respectively).

In conjugate [I], the pharmaceutical compound whose residue is represented by A-Co- is a pharmaceutical compound having a carboxy group in the molecule or a salt thereof, such as diclozenaf sodium, ibuprofen, flufenamic acid, mefenam. acids, anti-inflammatory agents such as aspirin, sodium salicylate, alclofenac, naproxen, flurbiprofen, ketoprofen, fuenbufuene, indomethacin, clidanac, sulindac, antitumor agents such as melphalan, methotrexate, hormones such as calcitonin, insulin-like growth factor summer Examples include agents.

The conjugates of this invention and their salts can be used depending on the type of spacer that binds the pharmaceutical compound and the diphosphonic acid derivative.
Each can be manufactured by the following method.

Production method 1 [I[] Production method 2 [1[] [IbCo production method 3 [■] [In the formula, A-Co- and n each have the same meaning as before, and OR1 is a protected hydroxy group, Y means a halogen atom. Raw materials [1] and [■] for the above production method can be produced by the following production method.

Jutoyo A [IK] NH(C1() R 22n 2 Hydrolysis ----→A-Co-N)t(CI(2)nCOOI
([■co (in the formula, A-CO-1Y, OR1 and n each have the same meanings as before, and R2 means a lower alphkyl dicarbonyl group) Each of the above definitions will be explained in detail below.

Examples of protected hydroxy groups include lower alkoxy groups such as methoxy, ethoxy, propoxy, isopropoxy, butoxy, secondary butoxy, isobutoxy, tertiary butoxy, pentyloxy, neopentyloxy, hexyloxy, etc. More preferred examples include 01-C4 alkoxy groups.

Halogen atoms include chlorine, bromine, iodine and fluorine.

Examples of the lower alkoxycarbonyl group include methoxycarbonylpoxycarbonyl, impropoxycarbonyl, butoxycarbonyl, and tertiary butoxycarbonyl.

Examples of the salt of the conjugate [1] include organic acid salts such as acetate, maleate, tartrate, methanesulfonate, benzenesulfonate, and toluenesulfonate, or hydrochloride, hydrobromide, and sulfuric acid. Acid addition salts such as inorganic acid salts such as salts and phosphates, metal salts such as sodium salts, potassium salts, calcium salts, magnesium salts, etc., organic amine salts such as ammonium salts, trimethylamine salts, triethylamine salts, dicyclohexylamine salts, etc. can be mentioned.

Here, the conjugates [Ia] to [I cl are included within the scope of the conjugate [I], so these conjugates [1a] to [
Regarding the salt of [IC], reference can be made to the salts exemplified for the above-mentioned conjugate [I].

The method for producing the target compound [1] and its salt will be explained in detail below.

Production method 1 Target compound [Ia] and its salt are pharmaceutical compound [1[
] or a salt thereof can be reacted with compound [III], and the resulting compound [IV] can be produced by subjecting the resulting compound to an elimination reaction of the hydroxy protecting group.

For suitable salts of pharmaceutical compound [II], reference can be made to the salts exemplified for conjugate [I] above.

The reaction of transferring compound [111] to pharmaceutical compound [I[] or a salt thereof to obtain compound [IV] is usually carried out using water, methanol, ethanol, propatool, tetralin, tetrahydrofuran, acetonitrile, dioxane, chloroform, toluene, The reaction is carried out in a solvent such as dimethylformamide or dimethyl sulfoxide, but the reaction can be carried out in any other organic solvent as long as it does not adversely affect the reaction.

Although the reaction temperature is not particularly limited, the reaction is usually carried out at room temperature or under heating.

This reaction can be performed with, for example, alkali metal hydrides such as sodium hydride and potassium hydride, alkaline earth metal hydrides such as calcium hydride and magnesium hydride, and alkali metal hydroxides such as thorium hydroxide and potassium hydroxide. alkali metal carbonates such as sodium carbonate and potassium carbonate; alkali metal hydrogen carbonates such as sodium hydrogen carbonate and potassium hydrogen carbonate; alkali metal fluorides such as potassium fluoride and cesium fluoride; sodium methoxide and sodium ethoxide. , alkali metal alkoxides such as potassium tert-butoxide, trialkylamines such as trimethylamine and triethylamine, picoline, 1.5-
Diazabicyclo [4.

3,0]non-5-ene, 1,4-diazabicyclo[
2,2.2 co-octane, 1,5-diazabicyclo[5
, 4.0] undecene-5 and the like in the presence of an inorganic or organic base.

The reaction of removing the hydroxy protecting group of compound [IV] to obtain the target compound [1a] or a salt thereof is carried out by conventional methods such as hydrolysis and reduction.

Hydrolysis is preferably carried out in the presence of a base, an acid including a Lewis acid, or a halosilane compound.

Examples of the base include the above-mentioned inorganic bases and organic bases, and examples of the acid include organic acids such as diric acid, acetic acid, propionic acid, trichloroacetic acid, and trifluoroacetic acid, as well as hydrochloric acid, hydrobromic acid, sulfuric acid, Examples include inorganic acids such as hydrogen chloride and hydrogen bromide.

Examples of the halosilane compound include halotri(lower)alkylsilanes such as iodotrimethylsilane and bromotrimethylsilane.

Note that the elimination reaction using a Lewis acid such as a trihaloacetic acid such as trichloroacetic acid or trifluoroacetic acid is preferably carried out in the presence of a cation scavenger such as anisole or phenol.

This reaction is usually carried out in water, an alcohol such as methanol or ethanol, methylene chloride, acetonitrile, chloroform, tetrachloromethane, tetrahydrofuran, or a mixed solvent thereof, but any other solvent may be used as long as it does not adversely affect the reaction. The reaction can also be carried out in a solvent. Note that a liquid base, acid, or halosilane compound can also be used as a solvent.

The reaction temperature is not particularly limited, and the reaction is usually carried out under cooling or heating.

On the other hand, reduction methods applied to the elimination reaction of hydroxy protecting groups include chemical reduction and catalytic reduction.

Suitable reducing agents that can be used for chemical reduction include, for example,
Metals such as zinc and iron or metal compounds such as chromium chloride and chromium acetate, and organic or inorganic acids such as acetic acid, propionic acid, trifluoroacetic acid, p-toluenesulfonic acid, hydrochloric acid, and hydrobromic acid. It is a combination of

Suitable catalysts used in the catalytic reduction include, for example, platinum catalysts such as platinum plates, platinum sponges, platinum black, colloidal platinum, platinum oxide, platinum wire, palladium sponges, palladium black, palladium oxide, palladium-backed elements, and colloidal palladium. , palladium catalysts such as palladium-barium sulfate and palladium-barium carbonate, nickel catalysts such as reduced nickel, nickel oxide, and Raney nickel, reduced cobalt, cobalt catalysts such as Raney cobalt, reduced iron, iron catalysts such as Raney iron, reduced copper, Conventional catalysts such as copper catalysts such as Raney copper, Ullmann steel, etc.

Elimination reactions by chemical reduction are usually carried out in commonly used solvents that do not adversely affect the reaction, such as water, methanol, ethanol, propatool, N, N-', and methylformamide, or in a mixture of these solvents. It will be done. Note that if the acid that can be used for chemical reduction is in liquid form, it can also be used as a solvent.

In addition to the above-mentioned solvents, examples of the solvent used in the catalytic reduction include conventional solvents such as diethyl ether, dioxane, and tetrahydrofuran, and mixed solvents thereof.

The reaction temperature of these reactions is not particularly limited, and the reactions are usually carried out under cooling or heating.

Production method 2' The target compound [Ib] and its salt are the pharmaceutical compound [1[
] or a salt thereof with compound [V] or a reactive derivative thereof at an amine group, or a salt thereof, and compound [VI] obtained by reacting the compound [VI] can be produced by subjecting the compound [VI] to an elimination reaction of the hydroxy protecting group.

Regarding the salt of compound [V], reference can be made to the salts exemplified for conjugate [I].

As the reactive derivative at the amino group of compound [V], commonly used derivatives that can be used in the amidation reaction, such as Schiff base-type imino produced by the reaction of compound [V] and a carbonyl compound or its enamine-type tautomer Examples include silyl derivatives produced by the reaction of compound [V] with silyl compounds such as triphthylsilylacetamide and bis(trimethylsilyl)acetamide, and derivatives produced by the reaction of compound [v] with the trichloride groove or phosgene. It will be done.

The reaction to obtain compound [VI] by reacting pharmaceutical compound [11] or its salt with compound [V] or its reactive derivative at the amine group or its salt is usually carried out using water, methanol, ethanol, propatool, tetralin, or tetrahydrofuran. , dioxane, chloroform, toluene,
The reaction is carried out in a commonly used solvent such as dimethylformamide, dimethylsulfoxide, or dichloromethane, but the reaction can also be carried out in any other organic solvent as long as it does not adversely affect the reaction.

Although the reaction temperature is not particularly limited, the reaction is usually carried out under cooling, room temperature, or heating.

This reaction is performed on N,N-dicyclohexylcarbodiimide, phosphorus oxychloride, trichloride, pentachloride, thionyl chloride, oxalyl chloride, 1-(p-'70-benzenesulfonyloxy)-6-chloro -IH-benzotriazole, chloride (chloromethylene) produced by the reaction of dimethylformamide with thionyl chloride or phosgene
It is preferably carried out in the presence of a conventional condensing agent such as the so-called Vilsmeier reagent, such as a compound formed by the reaction of dimethylammonium, dimethylformamide and phosphorus oxychloride.

The reaction of removing the hydroxy protecting group of compound [VI] to obtain the target compound [Ib] or a salt thereof can be carried out in the same manner as in Production Method 1.

The target compound [Ic] and its salt are obtained by hydroxy-protecting the compound [■] obtained by reacting the compound [■] or its salt with the compound [V] or its reactive derivative at the amine group or its salt. It can be produced by subjecting it to a group elimination reaction.

As the salt of the compound [■], the salts exemplified for the conjugate [I] can be referred to.

These reactions can be carried out in the same manner as in Production Method 2.

The compounds obtained in Production Methods 1 to 3 can be converted into salts as described above by conventional methods.

Next, the raw material compound [111] and the raw material compound [■]
The method for producing the compound and its salt will be explained in detail below.

Manufacturing company Raw material compound [I[[] is produced by reacting compound [IX] or its reactive derivative at the carboxy group or its salt with compound [V] or its reactive derivative at the amino group or its salt. be able to. For the salt of compound [IX], the conjugate [I]
Reference may be made to the salts exemplified for.

Examples of reactive derivatives at the carboxy group of compound [IX] include acid halides, acid anhydrides, activated amides, activated esters, and the like.

Suitable examples of such reactive derivatives include acid chlorides, acid azides, substituted phosphoric acids such as dialkyl phosphoric acid, phenyl phosphoric acid, and aliphatic carboxylic acids such as pivalic acid, acetic acid, trichloroacetic acid, etc. mixed acid anhydrides, symmetrical acid anhydrides, activated amides with imidazole, triazole or dimethylpyrazole, N-hydroxysuccinimide,
N-hydroxyphthalimide or 1-hydroxy-6
Examples include activated esters with =chlorobenzotriazole.

This reaction can be carried out in the same manner as the method for obtaining compound [VI] described in Production Method 2.

1 Sumitomo 1 Raw material compound [■ and its salt are obtained by hydrating the compound [XI] obtained by reacting the pharmaceutical compound [I] or its salt with the compound [X] or its reactive derivative at the amino group or its salt. It can be produced by subjecting it to a decomposition reaction.

Regarding the salt of compound [X], reference can be made to the salts exemplified for conjugate [I].

The reaction to obtain compound [XI] by reacting pharmaceutical compound [II] or a salt thereof with compound [X] or a reactive derivative thereof at an amino group or a salt thereof is the same as the method for obtaining compound [VI] in Production method 2. It can be carried out in the same manner, and the reaction of hydrolyzing compound [XI] to obtain starting compound [2] or a salt thereof can be carried out in the same manner as the hydrolysis reaction in Production Method 1.

If the target compound [I] has one or more stereoisomers such as optical isomers or geometric isomers based on asymmetric carbon atoms, double bonds, etc. in the molecule, such stereoisomers of the target compound [I] All isomers and mixtures thereof are also included within the scope of the target compound [I].

The conjugate [I] of the present invention is prepared by a conventional method into a solubilized preparation, an emulsion preparation, a liposome preparation, etc., and then administered by injection (for example, intravenous injection, intramuscular injection, intraarticular injection, etc.)
, administered in vivo by oral administration, rectal administration, etc.
Administration by injection is most preferred.

After the conjugate [I] of this invention has been administered to a living body, it is selectively taken up into the bone tissue, maintains the concentration in the bone at a high level for a long period of time, and has the ability to interact with diphosphonic acid derivatives in the bone tissue. The bond is broken and the original drug compound is gradually converted, allowing the concentration of the original drug compound in the bone tissue to be maintained over a long period of time.

In addition, in pharmaceutical compounds having two or more carboxyl groups in the molecule or their salts, if the diphosphonic acid derivative is not bound to the pharmacologically active carboxyl group via a spacer, the conjugate itself However, even if the conjugate is a pharmaceutical compound or its salt having one carboxy group in its molecule, the conjugate obtained by this invention may itself have sufficient pharmacological activity. .

Since the conjugate [I] of this invention is selectively taken up into bone tissue, it can be used as an anti-inflammatory agent to treat rheumatoid arthritis,
It can be used to treat osteoarthritis, low back pain, etc., and when applied to bone disease therapeutics (e.g. insulin-like growth factor I, etc.), it can be used to treat bone diseases (e.g. osteoporosis, budget bone disease, osteolysis, etc.) It can be used for the treatment of cancer, and if applied as an anticancer drug,
It can also be used to prevent cancer from metastasizing to bones.

In addition, once the conjugate [I] is distributed in the bone tissue, by gradually releasing the pharmaceutical compound from there, the blood concentration can be maintained for a long period of time, so a trace amount of blood concentration can be maintained. However, it can also be applied to pharmaceutical compounds (eg hormones, bioactive polypeptides, etc.) which are reflected in improved treatment.

The conjugate [I] of this invention and its salt are usually 10 to 1
A unit dose of 000 mg is administered once every 1 to 15 days, but the dose can be adjusted as appropriate depending on the type of pharmaceutical compound, patient's age, weight, symptoms, administration method, use in combination with other drugs, etc.

[Effect of the invention] The effects of the present invention will be explained below using test examples.

s1u [1 so (i) In the following test examples, rcarboxyfluorescein J
refers to the above-mentioned compound.

(i) (Conjugate obtained in Example 1-(i) below) In the following test examples, "conjugate obtained in Example 1-(i) below" refers to the above-mentioned iso-scenic compound. Say something.

An aqueous solution (fi degree: 200 μM) of the conjugate obtained in Example 1-(i) below was added to 1 L star method rat plasma 0.95 mu.
, 05ml was added and cultured at 37°C. After adding the aqueous solution of the conjugate, Q at 1, 2, 4, and 6 hours, respectively.
, 1 mQ samples were collected. The collected sample was diluted with distilled water Q, 4mQ, and immediately filtered through a column guard (Millibore 5JHVOO4NS, 0.45p), and the concentration of the remaining conjugate and the concentration of carboxyfluorescein produced were determined by high performance liquid chromatography. .

The above test results show that the conjugate of the present invention is gradually converted to carboxyfluorescein in plasma.

Test Example 2 pH 7.4 of the conjugate obtained in Example 1-(i) of Ying J Nii Postscript for 1 dust of Keicheng Jibori
Phosphate buffer solution (a degree: 1.63 mg/mQ
) and carboxyfluorescein in p) 17.4 phosphate buffer solution (A degree: 0.94 mg/ml), respectively, and 5 ml of SD male and female rats weighing approximately 2
50 g) into the tail vein, and those who received the conjugate were administered at 1 hour, 1, 2, 4, 7, and 14 days after administration, and those who received carboxyfluorescein were administered at 1 hour and 14 days after administration. After one day had elapsed, each femur was collected, the pieces of meat were removed and freeze-dried, and the dry weight was measured.

The femur was then treated with 6N hydrochloric acid saturated with sodium chloride.
Add 11IQ per 100mg of femur and leave at room temperature for 4 hours. After shaking and centrifugation, in-amyl alcohol 1.25111 is added to the femoral bone solution 1- for extraction. Divide the organic layer into two parts of 0.5 mQ, add 1.5-m of pH 7.4 phosphate buffer to one organic layer Q, and 5 mQ, shake and centrifuge to determine the conjugate concentration in the aqueous layer. Measured by high performance liquid chromatography.

In the other organic layer, 0.5 - IN sodium hydroxide was added.
．． After adding 5ml of the mixture and leaving it for 30 minutes at room temperature, it was shaken and centrifuged, and the total carboxyfluorescein concentration in the aqueous layer was measured by high performance liquid chromatography. Carboxyfluorescein concentration in the femur was calculated according to the following formula.

(Concentration of carboxyfluorescein) - 1 Concentration of carboxyfluorescein - (Concentration of conjugate) From the above test results,
It can be seen that the conjugate of this invention exhibits a significantly higher bone concentration than when carboxyfluorescein is administered. It can be seen that the bonds of the conjugate are gradually broken in the bone and converted to carboxyfluorescein.

In addition, the concentration of carboxyfluorescein in the bone produced is significantly higher than the carboxyfluorescein concentration in blood shown in Test Example 3 below, and the concentration is approximately 1000 times higher than that of carboxyfluorescein in blood.
It is found to last up to a day.

Kirani, bone concentration 1.60% dose 1 hour after conjugate administration
/femur contains 62% of the dose when converted into whole rat bone.
(Similarly, the bone concentration 1 hour after administration of carboxyfluorescein is 0.03 dose%).
/femur is equivalent to 1.0% of the administered dose when converted to whole rat bone.
(1% is distributed in bones) It can be seen that the conjugate of this invention is extremely easily distributed in bones.

Test Example 3 Monkisei Standing Soil 11 Test Method Pi(7,
4-phosphate buffer solution (concentration: 8.14 mg/mfl) and carboxyfluorescein pH 7, 4-phosphate buffer solution (a degree: 4.7 mg/mfl), respectively.
, 5 mQ was injected into the tail vein of SD male rats (weight 250 g).
minutes, 1 hour, 4 hours, 1 day, 2 days, 4 days, 7 days and 1
At the end of 4 days, the carboxyfluorescein-administered animals were treated at 5 minutes, 15 minutes, 30 minutes, 1 hour, and 2 hours after administration.
Blood was collected from the jugular vein after a period of time, and after centrifugation, plasma was removed.
．． 1 mQ was collected, and g of distilled water and ImQ were added to prepare a sample. Carboxyfluorescein concentration in blood was measured by high performance liquid chromatography.

From the above test results, when the conjugate of this invention is administered by injection, carboxyfluorescein remains in the blood for 14 days, compared to when carboxyfluorescein is administered by injection (calculated half-life of about 20 minutes). It can be seen that the blood concentration is significantly sustained.

The conjugate of flufenamic acid and a diphosphonic acid derivative obtained in Example 6-(1) described later in the 1st Kudo and the physiological saline solution of flufenamic acid were each administered at 40 μmol/k to ICR male mice (body weight: approximately 40 g).
g and 100 μmol/kg by tail vein injection,
Femurs were harvested at 15 minutes, 30 minutes, 1.2.4 and 24 hours after administration. The drug concentration in the collected samples was measured by high performance liquid chromatography.

(Eichen Souse) The above test results show that the conjugate of the present invention is more easily incorporated into bone tissue than the original pharmaceutical compound.

[Example] The present invention will be explained in more detail below with reference to Production Examples and Examples.

11 Counterfeit (aminomethylene) bis(phosphonic acid) tetraether (
To a solution of 2,25 g) in dichloromethane (20 mJl) was added α-bromoacetic acid (0,973 g) under water cooling.
Furthermore, N,N-dicyclohexylcarbodiimide (1,
Add a solution of 45g) in dichloromethane (5ml) and
After stirring at ℃ for 1 hour, it is stirred at room temperature for 20 hours. After filtering off the resulting dicyclohexylurea, the solvent is distilled off. Water and ethyl acetate were added to the residue for extraction, the organic layer was dried over magnesium sulfate, the solvent was distilled off, and (α-
Tetraethyl bromoacetamitomeprene bis(phosphonate) (2.42 g) is obtained.

NMR (DMSO-ds, 8)' 1.25 (1
2H1t), 3.95-4.20 (10H, m),
4.65-4.95 (LH, m>j11) Also, in the same manner as in Production Example 1, the following compounds were obtained.

(1) (6-promohexanoamidemethylene)bis(phosphonic acid)tetraethyl NMR (DMSO-d6.δ): 1.15-1.9
0 (188, m), 2.26 (2H, t), 3.
52 (2H, t>, 3.85~4.20 (8H,
m).

4.70-5.10 (IH, m) (2) (11-bromoundecanoamidomethylene)bis(phosphonic acid)tetraethyl NMR (DMSO-d6.8) i 1.10-1.
85 (18H, m), 2.22 (2H, t), 3
．． 55 (2H, t), 4.0=4.20 (8H,
m).

4.65-4.95 (IH, m) Carboxyfluorescein (1) was added to a solution of α-aminoacetate methyl hydrochloride (0,975 g) and triethylamine (1,0811111) in dry dimethylformamide (20 ml). 3',6'-dihydroxy-3-oxose 10[phthalane-1,9'-xanthine]-6-
Carboxylic acid and 3′,6′-dihydroxy-3
-oxo-spiro[phthalane-1,9'-xanthine]
-5-carboxylic acid mixture) (2,63 g) was added under water cooling, and N,N-dicyclohexylcarbodiimide (1,77 g) was added in dry dimethylformamide (5 m
Q) Add the solution and stir at 0°C for 1 hour, then at room temperature for 22 hours. After the generated dicyclohexylurea is filtered off, the solvent is distilled off. Ethyl acetate (5
After washing with 0.5N hydrochloric acid (3QmQ) twice and drying over magnesium sulfate, the solvent was distilled off. The resulting residue was poured into a silica gel column (80 g) and eluted with a mixture of chloroform and methanol (50:1). The solvent was distilled off from the eluate to obtain methyl α-(3
',6'-dihydroxy-3-oxo-spiro[phthalan-1,9'-xanthene-6-carboxamide) acetate and methyl α-(3',6'-dihydroxy-3-oxo-spiro[phthalan-1, Mixture of 9'-xanthene-5-carboxamide) acetate (1,
1 g) is obtained.

(i) Ethanol (30ml) is added to the compound (1.1g) obtained in (i) above, and IN sodium hydroxide (Ltmu) is added under water cooling, followed by stirring at 0°C for 4 hours. The pH was adjusted to 7 with 10% citric acid, ethanol was distilled off, and the pH was adjusted to 3 with 10% citric acid while cooling with water. The precipitate was collected by filtration, dried, and recrystallized from a mixture of methanol and diisopropyl ether to give α-(3',6'-dihydroxy-3-oxo-spiro[phthalane-1,9'-
xanthine]-6-carpoxamide]acetic acid and α-(
3',6'-dihydroxy-3-oxo-spiro[phthalane-1,9'-xanthene-5-carboxamide)
A mixture of acetic acid (0.92 g) is obtained.

NMR (DMSO-da, 8): 3.95-4-1
0 (2H9m>, 6-50~6.75 (6H, m)
, 7.4 (1 to 8.55 (3H, m)) Example 1 (i) To a solution of carboxyfluorescein (0,158 g) in dry metherformamide (2 mQ) was added triethylamine (71JJQ) and the compound obtained in Production Example 1. (α
-Tetraethyl bromoacetamitomeprene bis(phosphonate) (0.28 g) is added and stirred for 2 hours at 70"C. The resulting reactive mixture is poured into a mixture of 0.5N hydrochloric acid and ice water (100 mA). , ethyl acetate (50 m
Extract twice with Q), dry over magnesium sulfate, and evaporate the solvent. The resulting residue is subjected to adjusting bed chromatography, extracted with a mixture of chloroform and methanol (4:1), and the solvent is distilled off from the extract. Ethyl acetate (30 mQ) was added to the residue, washed with 50% brine, dried over magnesium sulfate, and the solvent was distilled off to give [α-(3'
, 6'-dihydroxy-3-oxo-spiro[phthalane-1°9'-xanthine]-6-carbonyloxy)acetamidomethylene]bis(phosphonic acid)tetraethyl and [α-(3',6'-dihydroxy -3-oxo-
A mixture (0,216 g) of spiro[phthalane-1,91-xanthine]-5-carbonyloxy)acetamidomethylenecobis(phosphonate)tetraethyl is obtained.

NMR, (DMSO-ds, S)' 1.05-1
．． 40 (12H), 4.0-4.25 (8H, m
), 4.65-5.20 <3H, m), 6.50
~6.75 (6H, m), 7.40-8.50 (
3H,m>(i) The mixture obtained in (i>) above (0,2
Iodotrimethylsilane (0.31 mQ) was added to a dry acetonitrile (2 mA) solution of 0.1g) at -20°C, stirred from -20"C to 0°C for 30 minutes, and then stirred at 0°C.
Stir at C for 2 hours. The solvent was distilled off, and the residue was mixed with water (20 m
1 l) and dichloromethane (201 m),
Stir for 30 minutes at O'C. The aqueous layer was dichloromethane (1
2111') three times, and the water is distilled off. Water and sodium acetate (0.08 g) were added to the resulting residue, and 4
Distill at 0"C.

Crystallized from a mixture of water and ethanol, [α-(3
',6'-dihydroxy-3-oxo-spiro[phthalan-1,9'-xanthine]-6-carbonyloxy)
disodium salt of [acetamidomethylene]bis(phosphone #) and [α-(3′).

6'-dihydroxy-3-oxo-spiro[phthalane-
A mixture (0,127 g) of the disodium salt of 1,9'-xanthencho-5-carbonyloxy)acetamidomethylenecobis(phosphonic acid) is obtained.

NMR (D20.S)' 3.95~4-25 (
2) 19m), 6.60-7-40 (6H, m),
7.50 to 8.35 (3H, m>11) The following compounds are obtained in the same manner as in Main Example 1.

(1) Disodium salt of [6-(3',6'-dihydroxy-3-oxo-spiro[phthalane-1,9'-xanthine]-6-carponyloxy)hexanoamidemetelenchobis(phosphonic acid) and [6-(3',6'-
Dihydroxy-3-oxo-spiro[phthalane-1,9
'-xanthine]-5-carbonyloxys(phosphonic acid) disodium salt mixture NMR (
DMSO-da. 8) 't. 3o-t. 95
(6) 1, m), 2. 30 (2H.t), 4.1
5-4.50 (2H, m), 6.60-7.35 (
6H. m), 7.45-8.35 (3H.m) (2
)
Mixture of disodium salts of (3',6'-dihydroxy-3-oxo-spiro[phthalan-1,9'-xanthine]-5-carbonyloxybis(phosphonic acid) NMR
(ND3,S)' 1.0-1. 95 (6H,
m), 2.32 (2H.

t), 4.05-4.50 <28, m), 6.6
0-7.25 (6H, m).

7, 40-8.50 (3H, m) Example 3 (1) Carboxyfluorescein (0.376g)
in dry dimethylformamide solution (31d) of (
aminomethylene) bis(phosphonic acid) tetraether (0
．． 32g) and N. A solution of N-dicyclohexylcarbodiimide (0.22 g) in dry dimethylformamide was added at 0°C, stirred at the same temperature for 1 hour, and further stirred at room temperature for 1 hour.
Stir for 6 hours. Dicyclohexylurea is filtered off, and the solvent is distilled off. The resulting residue was added with 4% sodium hydrogen carbonate (20+1111) and ethyl acetate (2QmQ
) and shake. Dilute the water reservoir with ethyl acetate (20mQ
), then adjust the pH to 2 by adding IN@acid and extract with ethyl acetate. Separate the organic layer, dry with magnesium sulfate, and evaporate the solvent. The obtained residue was subjected to preparative layer chromatography (Merck 5717) and eluted with a mixture of chloroform and methanol (4:1).
Add ethyl acetate (3oma) and 50% saturated saline and shake. Separate the organic layer, dry with magnesium sulfate, and evaporate the solvent. Powdered by treatment with isopropyl ether to give (α-3′,6″-dihydroxy-
3-oxo-spiro[phthalane-1,9'-xanthine]-6-carpoxamidomedelene)bis(phosphonic acid)
Tetraethyl and (α-3′.

6'-dihydroxy-3-oxo-spiro[phthalane-
1.9'-xanthine]-5-carboxamidomethylene)bis(phosphonic acid)tetraethyl mixture (0.16
6g) is obtained.

NMR (DMSO-d6, δ) + 1.0-1.4
(12H.111>, 3.95~4, 25 (8H
．． m), 4.95-5.35 (IH.m>, 6.
5-6.8 (6H.a+), 7.3-8.6 (3H.
, m) (i) The mixture obtained in (i) above was prepared in Example 1.
- (α-3′,6′-dihydroxy-3-oxo-spiro[phthalan-1.9′-
xanthine]-6-carpoxamidomethylene)bis(phosphonic acid) disodium salt and (α-3'.6'-
Dihydroxy-3-oxo-spiro[phthalane-1.9
'-xanthene-5-carboxamidomethylene)bis(phosphonic acid) disodium salt mixture (0.095
g) is obtained.

NMR (D20.8): 6. 60-7. 35
(6)1. m), 7. 50-8. 35 (3H, m
) α-(3',8'- obtained in Production Example 3-(i)
Dihydroxy-3-oxo-spiro[phthalane-1,9
'-xanthine-6-carpoxamide) acetic acid and α
-(3'.6'-dihydroxy-3-oxo-spiro[
(α-3′.6′-dihydroxy- 3-oxo-spiro[phthalane-1.9'
-xanthene 6-carbonylglycylaminomethylene)bis(phosphonic acid)tetraethyl and (α-3'
．． 6'-dihydroxy-3-oxo-spiro[phthalane-1.

9′-xanthine]-5-carbonylglycylaminomethylene)bis(phosphonic acid)tetraethyl mixture (0
, 69g).

NMR(DMSO-d8)'L, (14,4
(12)1. m), 3.95~6°4.25 (10B, m), 6.5~6.8 (6H
, m), 7.3 to 8.6 (3H, m) (i) Using the mixture (0, 2 g) obtained in (1) above, process in the same manner as in Example 1-(i), (α−3'
, 6'-dihydroxy-3-oxosespiro[phthalane-1,9'-xanthene-6-carponylglycylaminomethylene)bis(phosphonic acid) disodium salt and ('a-3', 6'mi3', 6'-dihydroxy-pyro[phthalane-1゜9'-xanthene-5-
Carbonylglycylaminomethylene) bis(phosphone #
) is obtained (0.06 g).

NMR (D20,8)' 3.95-4.2
5 (2H9m), 6.60~7.35 (6H,m
), 7.50 to 8.35 (3H, m) Example 5 (i) Diclzenaf sodium (0,382 g) and (α-bromoacetamidomethylene)bis(phosphonic acid) tetraethyl obtained in Production Example 1 (0, so9g)
was added to dry dimethylformamide (4 ml+) and stirred at room temperature for 3 hours. Water (50111) and ethyl acetate (50 ml) are added for extraction, the organic layer is dried over magnesium sulfate, and the solvent is distilled off. The resulting residue was subjected to preparative layer chromatography and eluted with a mixture of chloroform and methanol (10:1) to give [2-(2,6-dichloroanilino)phenylacetoxyacetamidomethylenezbis(phosphone)]. Tetraethyl acid) (0.58 g) was obtained.

NMR (DMSO-d6.δ): 1.23 (L2
H,t), 3.90 (2)1゜s), 3.95
~4.18 <8H, m), 4.68 <2H
,s).

6.80-7.60 (6H, m) (i) [2-(2,6-dichloroanilino)phenylacetoxyacetamidomethylenezbis(phosphonic acid)tetraethyl(0 , 4 g) of dry dichloromethane (10 mQ) g solution, iodotrimethylsilane (4684) was added dropwise at 0°C. The mixture is stirred at 0°C for 3 hours, then methanol (511111) is added and further stirred at room temperature for 10 minutes. Extract by adding water (601d) and dichloromethane (6011111”),
Wash the aqueous layer three times with dichloromethane (50+1111).

The solvent is distilled off, the residue is dissolved in methanol, the insoluble materials are filtered off, and the solvent is distilled off. Sodium acetate (0,141 g) was added to the resulting residue, concentrated, and crystallized from a mixture of water and ethanol to give [2-(2,6-dichloroanilino)phenylacetoxyacetamidomethylene bis(phosphon Acid) Nori sodium salt (0.2 g) is obtained.

NMR (D20,f; )' 4.02 (2H,s
), 4.80 (2H,s).

6.90-7.55 (6H, m) [2-[(3-trifluoromethyl phenyl)amino]benzoyloxyacetamidomethylenecobis(phosphonic acid) disodium salt (0,23
5g).

NMR (D20.l; )' 4.0-4.66
(LH9m), 4.88 (2H3s), 6.96
(18,t), 7.30~7.61 (6H,m),
7.98 (IH, d), 8.40 (IH, d) (
2) Using ibuprofen (0,206 g), treatment was carried out in the same manner as in Example 5 to obtain the disodium salt of [α-methyl-4-(2-methylpropyl)benzeneacetoxyacetamid methylene bis(phosphonic acid)]. (0.08 g) is obtained.

NMR (D20.l; ) 'o, 85 (3H,
s), o, 89 (3H9s).

Claims (3)

[Claims] (1) General formula ▲ Numerical formula, chemical formula, table, etc. ▼ (In the formula, A-CO- is a residue of a pharmaceutical compound, R is -NH- or -O-, m is 0 or 1, and n is 1 to (each represents an integer of 10) A conjugate of a pharmaceutical compound and a diphosphonic acid derivative, and a salt thereof. (2) A conjugate of a pharmaceutical compound and a diphosphonic acid derivative and a salt thereof according to claim (1), wherein R is -O- and m is 1. (3) A conjugate of a pharmaceutical compound and a diphosphonic acid derivative and a salt thereof according to claim (1) or (2), wherein the pharmaceutical compound is an anti-inflammatory agent.