JPH0656794A - New pilocarpine derivative and its preparation - Google Patents

Abstract

PURPOSE: To obtain a new bispilocarpic acid ester deriv. having intraocular tension degradation action, excellent in cornea permeability, ensuring a slight adverse drug reaction to the body and eyes and an increased consent rate of a patient and useful in the treatment of glaucoma, etc.

CONSTITUTION: 1,2-Dibromoetane is dropped into a soln. of sodium pilocarpate in dimethyl sulfoxide by ≤0.5 mol per 1 mol sodium pilocarpate and they are reacted at room temp. for 72 hr. The reaction mixture is poured into distilled water and this liq. mixture is subjected to extraction with chloroform. The reaction product is recovered from the extracting liq., crystallized and converted into a bispilocarpate of formula I [A is a direct bond or (substd.)methylene]. This bispilocarpate is reacted with an acid of the formula RCO₂H (R is 1-4C alkyl, 3-6C cycloalkyl or phenyl) or its functional deriv. to obtain the objective bispilocarpic acid ester deriv. (salt) of formula II (Y is RCO; and W is a group of formula III) useful in the treatment of glaucoma, etc.

COPYRIGHT: (C)1994,JPO

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to a novel pilocarpine prodrug compound useful for the treatment of glaucoma, and in particular to a bispyrocarpine acid ester, a method for producing the above novel compound, and a pharmaceutical composition containing the novel compound. About things and their use.

[0002]

[Prior art]

(+)-Pilocarpine, i.e. (3S-cis) -3-ethyl-dihydro-4
-[(1-Methyl-1H-imidazol-5-yl) methyl] -2 (3H) -furanone is used to treat glaucoma and lowers intraocular pressure by increasing the flow of aqueous humor from the eye It is a drug that lets you. The action of lowering the intraocular pressure by pilocarpine is based on the ciliary muscle contracting effect that widens the angle of the anterior chamber, which is important from the viewpoint of outflow of aqueous humor, and promotes outflow of aqueous humor.

However, the reduction of intraocular pressure is not the only effect of pilocarpine in the eye. If the drug concentration is high enough, the contractile effect of pilocarpine on ciliary muscles is increased, thereby accommodating the lens to see at close range. Therefore, it is difficult for the patient to adjust their eyes to look further away, which is inconvenient for the patient. Pilocarpine also contracts the iris,
Reduce the pupil significantly. Apart from these effects, which are unnecessary from a medical point of view and are uncomfortable for the patient, pilocarpine also causes side effects in areas other than the eye. Such side effects include in particular increased salivation and increased bradycardia.

Conventionally, patients with glaucoma have locally administered pilocarpine as eye drops. However, when administered in this manner, only about 1% of the pilocarpine dose is absorbed by the eye and about 70% is absorbed in the bloodstream. In this case, the low absorption rate of pilocarpine into the eye is due to the following three main factors. 1) A drop of liquid rapidly flows off the surface of the eye. 2) Rapid absorption of pilocarpine into the bloodstream through the conjunctiva on the inner surface of the eyelid. 3) The corneal permeability of pilocarpine is low.

Pilocarpine is absorbed by the eye through the cornea. In the cornea, pilocarpine is first absorbed by the dense epithelial layer on the surface of the eye, which is rich in cell membrane lipids (fats). However, since pilocarpine is not very soluble in fat, it penetrates the corneal epithelium relatively little.
The corneal epithelium acts as a film that suppresses the absorption of pilocarpine, and at the same time, it forms an aqueous stroma of the cornea.
It acts as a reservoir that carries pilocarpine through the ma) and endothelium to the aqueous humor of the anterior chamber. From the aqueous humor of the anterior chamber,
Pilocarpine can easily reach its site of action, the ciliary muscle. The action time of pilocarpine in the eye is significantly short due to the conversion of part of pilocarpine to inactive pilocarpine acid and the rapid outflow of pilocarpine from the eye due to the circulation of aqueous humor and the blood circulation in the iris. Become.

The low absorption of pilocarpine into the internal organs of the eye and the short duration of action of pilocarpine administered to the eye makes treatment with pilocarpine difficult. In order to improve the action of pilocarpine and to prolong its action time, a relatively large amount of pilocarpine must be used. By using a relatively large amount, in the aqueous humor,
High levels of pilocarpine concentration in the iris and in the ciliary muscles cause a strong contraction of the pupil and an adaptation of the eye for near vision. Furthermore, increasing the dose of pilocarpine, a type of drug that is rapidly eliminated from the eye, is a relatively ineffective method of prolonging its action, so pilocarpine eye drops are used depending on the patient. It is administered 3 to 8 times a day. Such frequent administration of eye drops is inconvenient for the patient, especially if side effects occur in the eye after each administration. Also, large doses increase the absorption of pilocarpine into the blood circulatory system, thereby increasing the risk of other side effects.

In order to overcome the above-mentioned drawbacks associated with poor absorption of pilocarpine, attempts have been made to use prodrug derivatives of pilocarpine that have better absorption into the corneal epithelium. To increase absorption, these prodrug derivatives of pilocarpine must be more soluble in fat than pilocarpine. Moreover, their prodrug derivatives degrade as completely as possible within the corneal epithelium,
The pharmaceutically effective pilocarpine and the ineffective ingredient [pro-moiety] must be released. The extent of degradation in the cornea depends on the residence time of the prodrug derivative within the corneal epithelium and its rate of degradation. The residence time of the prodrug derivative in the corneal epithelium increases with increasing lipophilicity and decreasing diffusion coefficient.

To date, two prodrug derivatives of pilocarpine have been developed. Border
Discloses in US Pat. No. 4,061,722 prodrugs of pilocarpine based on quaternary ammonium compounds. Bundgaard et al., In European Patent Application 0106541, disclosed pilocarpic acid diesters, which are used to enhance ocular absorption. However, this pilocarpic acid diester has low solubility in water and irritation to the eyes.
It has some drawbacks. Also, many undesired by-products are generated from this diester, as compared to the active ingredient, pilocarpine itself.

[0009]

SUMMARY OF THE INVENTION The present invention relates to a novel bispyrocarpic acid ester, namely bispirocarpate, which is capable of largely solving the abovementioned problems or at least minimizing them. The prodrug derivative according to the present invention is a Bundgaard
Prodrugs, etc. have the same lipophilicity as those of the same prodrugs, and at least equivalently rapidly decompose into pilocarpine and promoiety, and promote the permeability of pilocarpine through the cornea to at least the same extent as prodrugs such as Bundgard. . Furthermore, derivatives such as Bundguard are pilocarpine 1
Whereas one promoiety per molecule is carried into the cornea, the bispirocarpeto derivatives of the present invention carry one promoiety per two pilocarpine molecules. Since the diffusion coefficient of this bispyrocarpate derivative in the corneal epithelium is smaller than that of the compound such as Bundgard, the undegraded bispyrocarpate derivative stays longer in the corneal epithelium. Therefore, the time left for complete degradation of this prodrug is longer. Moreover, the novel compounds of the present invention have better solubility and are therefore more suitable for preparing pharmaceutical compositions.

As described above, according to the present invention, since the drug can be gradually released from the cornea to the internal organs of the eye, the action time of pilocarpine can be effectively prolonged and, further, pilocarpine in the eye can be treated. Since the high concentration peak can be lowered, it has an important effect from the viewpoint of reducing the above-mentioned side effects.

[0011]

The novel prodrug derivative of pilocarpine of the present invention is a bispyrocarpic acid ester represented by the following general formula 1, that is, bispyrocarpate or an acid addition product thereof.

In the formula 1, A) Y is -C (= O) -R (R is an alkyl group having 1 to 4 carbon atoms,
A cycloalkyl group having 3 to 6 carbon atoms or a phenyl group), and W is

The group represented by: [Y is as defined above, A is a direct bond, or a hydroxyl group, a group represented by YO- (Y is as defined above) and 1 One or two methyl groups are substituted or unsubstituted methylene groups], or B) W is an alkyl group having 1 to 4 carbon atoms or 3 to 6 carbon atoms.
Is a cycloalkyl group of, and Y is

[Chemical formula 3] (B is a direct bond, or a methylene group or an ethylene group, and R ′ has the same meaning as W above)
Is.

The first embodiment [A) of the compound of general formula 1
In the case of]

Embedded image (Y and A are as defined above).

In the general formula 4, R is preferably a cycloalkyl group having 3 to 6 carbon atoms, or linear or branched propyl or butyl. A is preferably a direct bond or an unsubstituted methylene group.

A second embodiment of the compound of the general formula 1 [B)
In case of], the general formula 6

Embedded image (R ′ and B are as defined above).

In the general formula 6, R'is preferably a cycloalkyl group having 3 to 6 carbon atoms, or linear or branched propyl or butyl. B is preferably a direct bond or an unsubstituted methylene group.

In the above general formula 1, the number of carbon atoms is 1 to 4.
The alkyl group of is a linear or branched methyl,
Ethyl, propyl or butyl is advantageously used, preferably linear or branched propyl or butyl such as n-propyl, isopropyl, n-butyl, isobutyl, t-butyl.

In the compound of the general formula 4 and the compound of the general formula 6 in a preferred embodiment, R and R'are the same.

When R or R'is a cycloalkyl group,
3 to 3 carbon atoms such as cyclopropyl or cyclobutyl
It is a cyclic body of 6 and may be substituted with a methylene group.

The acid addition salt of the compound represented by the general formula 1 is as follows:
Preferred are pharmaceutically acceptable addition salts with non-toxic inorganic or organic acids. Examples of suitable acids include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and organic acids such as acetic acid, propionic acid, stearic acid, oxalic acid, malonic acid, succinic acid, glutaric acid, adipic acid, Mention may be made of maleic acid, fumaric acid, malic acid, tartaric acid, citric acid, ascorbic acid, benzoic acid, pamoic acid, sulphonic acids such as mesylic acid and tosylic acid.

Preferred compounds of the general formula 4 are O, O'-dicyclopropylcarbonyl (1,2-ethylene) bispyrocarbate, O, O'-dicyclobutylcarbonyl (1,2-ethylene) bis Pilocarpeto, O, O'-dicyclopropylcarbonyl (1,3-propylene) bispyrocarpate, O, O '
-Dicyclobutylcarbonyl (1,3-propylene) bispyrocarbate, O, O'-dicyclopropylcarbonyl (2-
Methyl-1,3-propylene) bispyrocarpate, O, O'-dicyclopropylcarbonyl (2,2-dimethyl-1,3-propylene) bispyrocarpate, O, O'-dipropionyl (1,2 3
-Propylene) bispyrocarpate, O, O'-dicyclopropylcarbonyl (2-hydroxy-1,3-propylene) bispirocarpate, O, O'-dicyclopropylcarbonyl (2
-Cyclopropylcarbonyloxy-1,3-propylene) bispyrocarbate, O, O'-dipivalyl (1,2-ethylene)
Bispyrocarpet, O, O'-dipivalyl (1,3-propylene) bispyrocarpate, O, O'-di (1-methylcyclopropylcarbonyl) (1,2-ethylene) bispyrocarpate, O, O '-Dicyclopentylcarbonyl (1,2-ethylene) bispyrocarbate, O, O'-dipropionyl (1,2-
Ethylene) bispyrocarpate, O, O'-diisobutyryl (1,2-ethylene) bispyrocarpate, O, O'-dipropionyl (2-hydroxy-1,3-propylene) bispyrocarpate, O, O'-dicyclohexylcarbonyl (1,2-ethylene) bispyrocarpate, O, O'-dicyclopentylcarbonyl (1,3-propylene) bispyrocarpate, O, O '
-Dicyclohexylcarbonyl (1,3-propylene) bispyrocarbate, O, O'-dibenzoyl (1,2-ethylene)
Bispirocarbate or O, O'-dibenzoyl (1,2-
Propylene) bispyrocarpate.

Preferred compounds of general formula 6 are O, O'-succinyl (diethyl) bispyrocarpate, O, O'-succinyl (diisopropyl) bispyrocarpate, O, O'-
Succinyl (di-t-butyl) bispyrocarbate, O, O'-
Succinyl (dicyclopropyl) bispyrocarbate,
O, O'-succinyl (dicyclobutyl) bispyrocarpate, O, O'-glutaryl (diisopropyl) bispyrocarpate, O, O'-glutaryl (di-t-butyl) bispyrocarpate, O, O ' -Glutaryl (dicyclopropyl) bispyrocarpate or O, O'-glutaryl (dicyclobutyl) bispyrocarpate.

The present invention also relates to a process for preparing the compounds of general formula 1.

According to the manufacturing method of the present invention: a) Formula 4

Embedded image [Y is —C (═O) —R (R is an alkyl group having 1 to 4 carbon atoms, a cycloalkyl group having 3 to 6 carbon atoms, or a phenyl group), and A is A direct bond, or
A hydroxyl group, a group represented by YO- (Y is as defined above) and a methylene group substituted or unsubstituted with a group selected from the group consisting of one or two methyl groups] The compound is prepared by the following general formula 5

Reacting a compound of formula (A is as defined above) with an acid of formula RCO ₂ H or a functional derivative thereof; or b) a compound of formula 6

Embedded image (B is a direct bond, or a methylene group or an ethylene group, and R ′ is an alkyl group having 1 to 4 carbon atoms or a cycloalkyl group having 3 to 6 carbon atoms). The compound is produced by the compound represented by the general formula 7.

Embedded image (R ′ is as defined above)

Reacting with a dicarboxylic acid of formula (B is as defined above) or a divalent acid derivative thereof,
And, if desired, consists of converting the compound obtained by the reaction to its acid addition salt.

In order to prepare the compound used as the starting material in the above preparation method a), the compound of the general formula 9

[Chemical 9] Pilocarpic acid of the general formula X-CH ₂ -A-CH ₂ -X '(X and
X ′ independently represents a hydroxyl group or a residue such as halogen, an acyloxy group, an alkylsulfonyloxy group or an arylsulfonyloxy group, and A is reacted with a compound as defined above.

In order to produce the compound used as the starting material in the above production method b), pilocarpic acid as defined above is prepared by the general formula RX (X is a hydroxyl group, or
Represents a residue such as halogen, an acyloxy group, an alkylsulfonyloxy group or an arylsulfonyloxy group, and R is as defined above.

In the above reaction, acid halides, anhydrides, alkyl sulfonates and aryl sulfonates are preferably used as the functional acid derivative. As the reaction solvent, a solvent inert to the reactants, for example, a hydrocarbon, a halogenated hydrocarbon, an ether,
Alternatively, a ketone or the like is used. Suitable hydrocarbons are aromatic hydrocarbons such as benzene and alkylbenzenes such as toluene and xylene. Suitable halogenated hydrocarbons include, for example, dichloromethane, chloroform,
And there is chlorobenzene. Examples of the ketone include acetone, ethyl methyl ketone, and isobutyl methyl ketone, and examples of the ether include diethyl ether, diisopropyl ether, dibutyl ether, 1,
4-dioxane may be mentioned. Other suitable solvents include dimethyl sulfoxide, dimethylformamide, acetonitrile and the like.

The reaction temperature is not critical, for example -1
It may be in the range of 0 ° C. to the boiling point of the solvent. Room temperature is preferably used. The reaction time can be selected from a wide range and is generally 12 to 72 hours, usually about 24 hours. For the reaction, it is advantageous to use an acid binder such as an alkali metal or alkaline earth metal carbonate or an organic base. Suitable metal carbonates include sodium carbonate and potassium carbonate, and the organic bases include pyridine and its homologue, 4- (dimethylamino).
Mention may be made of pyridine, quinoline and its homologues, N, N-dimethylaniline, and trialkylamines, preferably triethylamine. The above reaction can be carried out in a homogeneous solution or in a heterogeneous system with phase transfer.

The ester bond of the compound of the present invention can be formed by using known water-cleaving agents such as carbodiimide. In some cases, a known acid-catalyzed esterification reaction may be used.

The present invention further comprises a pharmaceutically acceptable adjuvant containing a compound according to the invention as an active ingredient.
djuvants) and excipients.

The pharmaceutical compositions according to the invention are prepared in a known manner by using the excipients and other adjuvants known in the art. Excipients are, for example, liquids,
It may be a suspension, emulsion, cream, ointment or the like. The pharmaceutical composition may be in the form of a solid pharmaceutical for insertion into the eye. As an example of a suitable form for administration, the compound according to the present invention is added to an appropriate concentration, for example, 0.1 to 4 in a sterile aqueous solution adjusted to an appropriate pH with a buffer or an acid or a base. % Ophthalmic solution, in which case the compounds according to the invention are preferably used in the form of water-soluble acid addition salts. The eye drop having a desired concentration is instilled preferably 1 to 3 times a day depending on the patient's condition.

Test Method The compounds of the present invention were tested by the following test method.

1) Lipophilicity The lipophilicity of the compound was determined by measuring the partition coefficient (P) of the compound at pH 7.40 and 5.00. The measurement was carried out in a 1-octanol-phosphate buffer mixture by measuring the concentration of the compound in the buffer phase before and after partitioning. From the results obtained, all compounds of the present invention were found to be pilocarpine [log P
= 0.01 (pH 7.40); -1.74 (pH
5.0)] is more lipophilic, and it can be seen that the lipophilicity of the derivative of the present invention can be adjusted by changing the substituent bonded to pilocarpic acid.

2) Enzymatic hydrolysis The half-life of the enzymatic hydrolysis of bispirocarpet according to the invention was determined in plasma / buffer mixture and / or in rabbit corneal homogenate.

A. Enzymatic hydrolysis in plasma / buffer mixture Half-life of enzymatic hydrolysis of bispyrocarpic acid diester in plasma / pH 7.40 phosphate buffer mixture (80% / 20%) at 37 ° C It was measured at. The logarithm of the residual diester was expressed as a function of time and the half-life T _1/2 was determined from the resulting plot.

From this result, in the buffer solution, stable diesters are decomposed under the influence of plasma esterase into intermediate products O, O'-dihydrogenbispyrocarpate and monopyrocarpate, which are It can be seen that the intermediate product chemically decomposes to pilocarpine which is active at physiological pH values. The degradation rate of the prodrug derivative is
It can be adjusted by changing the substituents attached to pilocarpic acid.

B. Enzymatic Hydrolysis in Corneal Homogenate The half-life of enzymatic hydrolysis of bispyrocarpic acid diester was determined by rabbit corneal homogenate (pH 7.40, 37 °).
It was measured in C). Corneal homogenate is 0.05M T
Prepared with ris buffer. Expressing the logarithm of the residual diester as a function of time, the half-life T
I decided _1/2 .

From these results, the esterase of rabbit cornea was found to use bispyrocarpic acid diester as an intermediate product O, O'-.
It can be seen that upon hydrolysis to dihydrogen bispyrocarpetes and monopyrocarpetes, these intermediate products chemically decompose to active pilocarpine at physiological pH values. The rate of degradation of the prodrug derivative can be adjusted by changing the substituents attached to pilocarpic acid.

3) Corneal permeability The corneal permeability of the novel bispirocarpate according to the present invention was measured in a diffusion chamber. In the diffusion chamber, the compound moved from the release phase (epithelial side) through the cornea to the receptive phase (endothelium side). Rabbit corneas were used in this test. Transmission coefficient (P
_app ) was calculated from the permeation rate.

From these results, the novel compound of the present invention was found to have pilocarpine (P _app = 2.77 cm / s × 1).
It is understood that it is possible to improve the corneal permeability of 0 ^-6 ).

Table 1 shows the test results of the compounds of the present invention obtained in the examples described below.

[Table 1]

The results show that the compound of the present invention is hydrolyzed into corneal pilocarpine, and that the group binding to pilocarpic acid can regulate the corneal permeability of the prodrug and the rate of pilocarpine formation. It should be noted that, for example, derivatives that release pilocarpine only at a low rate, due to moderately moderate chemical hydrolysis of intermediates formed from derivatives of the compounds of the invention, release pilocarpine more rapidly. Also has a low transmission coefficient.

CONCLUSION By changing the group attached to pilocarpic acid, the lipophilicity, enzymatic hydrolyzability of intermediates of the compounds of the invention,
Corneal permeability, chemical hydrolysis and thus production of active pilocarpine in the eye can be easily tailored for the purpose. Bispyrocarpic acid diesters are enzymatically hydrolyzed to produce bispyrocarpic acid esters and pilocarpic acid monoesters, which are chemically (naturally) hydrolyzed to pilocarpine.

The bispyrocarpic acid diester according to the present invention sends one promoiety to every two molecules of pilocarpine to the cornea, so that the production amount of promoiety is suppressed to the minimum. In this way, minimizing the amount of promoiety produced is intended to reduce eye irritation and pain caused by drugs.

[0044]

The bispyrocarpic acid diester of the present invention as an oil-soluble compound is effectively absorbed by the corneal epithelium, whereupon a suitable water-soluble intermediate [for example, O from the compound represented by the general formula 4 O'-dihydrogen (1,2-ethylene) bispyrocarpate or O-hydrogen (ethyl) bispyrocarpate] from the compound represented by the general formula 6 is rapidly produced, and this is a water-soluble compound in the epithelium. From the stroma to the site of action of the internal organs of the eye, producing persistent pilocarpine during the migration. Therefore, neither the prodrug derivative nor pilocarpine is excessively accumulated in the corneal epithelium, so that the irritation to the eye due to the excessive accumulation thereof can be prevented.

[0045]

By using the compound of the present invention,
It is possible to significantly solve the problems associated with drug therapy with pilocarpine [low biological availability, side effects on the body and eyes, frequent medication, and low patient acceptance rate]. it can. Due to the better corneal permeability, the dose of the compound of the present invention can be considerably lower, and the number of times of administration per day can be reduced, so that the side effects are reduced and the patient acceptance rate is improved. Drug therapy can be performed more effectively.

[0046]

EXAMPLES The following examples specifically illustrate the present invention, but do not limit the present invention in any way.

Equipment used Melting point measurement: Reichert Thermovar (Reichert The
rmovar) Refractive index measurement: Atago Illumina
tor) apparatus Measurement of pKa value: titration of pilocarpine derivative in water-ethanol (50% -50%) mixture Mass spectrometer: VG 70-250SE Measurement condition in electron impact ionization apparatus: electron energy: 70 eV (especially Unless otherwise noted) Ionization current: 500 μA Ionizer temperature: 150 ° C Sample holder temperature: 30 ° C → 500 ° C in 2 to 5 minutes Resolution: 10,000 Thermospray type mass spectrometer: VG Thermospray / Plasma spray (thermospray /
plasmaspray) VG Trio-2 quadropole type Beckmann 112 pump (The measurement conditions in thermospray ionization were optimized daily.) NMR spectrometer: Bruker AC 250 / Aspect ( Aspect) 3000 ¹ H / ¹³ C 5 mm dual probe CD ₃ OD 20 mg / ml δ ppm (tetramethylsilane = 0)

[0048]

Example 1 1302 mg (5.25 mmol) of pilocarpic acid sodium salt was added to 60 ml of dimethylsulfoxide , O, O'-dihydrogen (1,2-ethylene) bispyrocarpate ( Y = hydrogen, A = direct bond). 247 mg (1.31 mm) in the solution containing
ol) 1,2-dibromoethane was added dropwise to prepare pilocarpic acid monoester. This solution was mixed at room temperature for 72 hours and poured into 100 ml of distilled water. The mixture was extracted with chloroform in 100 ml portions each in two portions. Combine the chloroform extracts and add 100
It was washed with ml of distilled water, washed with 100 ml of 2% sodium bicarbonate solution, and further washed with 100 ml of distilled water. The chloroform extract was dried over calcium sulphate for 30 minutes, chloroform was evaporated under reduced pressure and the resulting bispirocarpate was crystallized with an ethyl acetate / ether mixture to give 332 mg (0.69 mmo of the title compound.
l) was obtained. Mp _{= 111~115 ° C pK a = 6.30} HR-MS- spectrum: m / e (relative intensity): 209
(8%), 208 (19%), 96 (30%), 95
(100%). NMR: δ 7.49 2H bs, 6.74 2H b
s, 4.30 4H m, 3.60 6H s, 3.54
4H m, 2.72 2H m, 2.56 2H m, 2.
51 2H m, 2.02 2H m, 1.68 4H m,
0.896 Ht. The sodium salt of pilocarpic acid, which is the starting material, can be obtained as follows. Pilocarpine hydrochloride (3.92 g; 16.00 mmol) was added to distilled water (4 m
1) and the resulting solution was cooled to about 0 ° C. To this solution, 18 ml of ice-cold 2M NaOH was added in 4 portions, and the mixture was allowed to stand at about 0 ° C for 1 hour. 5 ml of 1M
After neutralizing the excess NaOH with HCl, the solution was evaporated under reduced pressure. The residue was dissolved in 60 ml absolute ethanol and mixed at 60 ° C for 10 minutes. After cooling to 4 ° C, the insoluble content of NaCl was removed by filtration. The filtrate was evaporated under reduced pressure to give 3.93 g of pilocarpic acid sodium salt as a white, highly hygroscopic material. Similarly, the sodium salt of pilocarpic acid (10
21 mg; 4.11 mmol) and 1,3-dibromopropane (208 mg; 1.03 mmol) gave O, O'-dihydrogen (1,3-propylene) bispyrocarbate. The yield was 330 mg (0.70 mmol). Similarly, the sodium salt of pilocarpic acid (1674 mg;
6.75 mmol) and 1,3-dibromo-2-hydroxypropane (341 mg; 1.69 mmol) can give O, O′-dihydrogen (2-hydroxy-1,3-propylene) bispyrocarbate. O, O'-dihydrogen (2-hydroxy-1,3-propylene) bispyrocarbate is a sodium salt of pilocarpine acid and epichlorohydrin (2:
It can also be obtained from 1).

[0049]

Example 2 O, O'-dicyclopropylcarbonyl (1,2-ethylene) bis
Pyrocarbate (Y = cyclopropylcarbonyl; A = direct bond) of 540 mg (1.13 mmol) in toluene (150 ml)
O, O'-dihydrogen (1,2-ethylene) bispyrocarpate (see Example 1) and 1866 mg (13.5 mmo)
945 mg in a mixed solution containing 1) potassium carbonate.
Cyclopropylcarbonyl chloride (9.64 mmol) was added dropwise over about 24 hours to give the title compound. The mixture was stirred for 24-72 hours. 2% sodium bicarbonate solution (150 m
1) was added and the resulting mixture was stirred at room temperature for 3 hours.
The resulting layers were separated and the toluene phase was washed twice with water (2 x 15
(0 ml), washed with calcium sulfate for 30 minutes and evaporated under reduced pressure to give O, O'-dicyclopropylcarbonyl (1,2-ethylene) bispyrocarpate. Yield is 3
It was 04 mg (0.49 mmol). pK _a = 5.70 HR-MS- spectrum: m / e (relative intensity): 614
[M ^+. ] (1%), 408 (11%), 393 (10
%), 321 (7%), 320 (29%), 307 (9
%), 291 (16%), 209 (18%), 208
(11%), 207 (78%), 163 (29%), 1
62 (14%), 121 (47%), 113 (52
%), 96 (38%), 95 (100%). HR-MS: Molecular weight = 614.3299260 (measured value) 614.3315650 (calculated value) NMR: δ 7.56 2H bs, 6.78 2H b
s, 4.35 4H m, 4.08 4H m, 3.63
6H s, 2.69 4H m, 2.50 2H m, 2.
33 2H m, 1.69 4H m, 1.61 2H m,
1.10 6H t, 0.92 6H t, 0.90 8H
m.

[0050]

Example 3 O, O′-dipropionyl (1,2-ethylene) bispyrocarpe
Doo (Y = propionyl, A = direct bond) by the method described in Example 2 O, O'dihydrogen (1,2-ethylene) bispilocarpate (300 mg; 0.63 mmol) (see Example 1) Propionyl chloride (464mg; 5.02mmo
The title bispirocarpet was obtained from 1). Yield is 2
It was 43 mg (0.41 mmol). pK _a = 6.00 HR-MS- spectrum: m / e (relative intensity): 591
(5%), 590 [M ^+. ] (5%), 503 (7%),
397 (19%), 396 (85%), 395 (8)
%), 382 (20%), 381 (95%), 309
(11%), 209 (18%), 195 (45%), 1
21 (18%), 96 (78%), 95 (100%). HR-MS: molecular weight = 590.36363040 (measured value) 590.3315650 (calculated value) NMR: δ 7.61 2H bs, 6.78 2H b
s, 4.33 4H m, 4.08 4H m, 3.62
6H s, 2.68 4H m, 2.49 2H m, 2.
32 2H q, 1.68 4H m, 1.10 6H t,
0.906 Ht.

[0051]

Example 4 O, O'-dicyclobutylcarbonyl (1,2-ethylene) bispi
Locarpate (Y = cyclobutylcarbonyl, A = direct bond; fumarate) By the method described in Example 2, O, O′-dihydrogen (1,
2-ethylene) bispyrocarpate (470 mg; 0.9
8 mmol) (see Example 1) and cyclobutylcarbonyl chloride (931 mg; 7.86 mmol) to give the title bispyrocarpate. The fumarate was obtained by dissolving the obtained bispyrocarbate in toluene, adding a solution of fumaric acid in 2-propanol, and precipitating with petroleum ether. The yield was 810 mg (0.82 mmol). Melting point = 49 to 51 ° C. (fum.) HR-MS-spectrum: m / e (relative intensity): 642
[M ^+. ] (4%), 529 (6%), 436 (10
%), 423 (22%), 422 (100%), 421
(13%), 408 (18%), 407 (85%), 3
35 (10%), 221 (52%), 209 (12
%), 121 (13%), 96 (86%), 95 (50
%). HR-MS: Molecular weight = 642.3648680 (measured value) 642.3628651 (calculated value) NMR: δ 8.50 2H bs, 7.23 2H b
s, 6.73s (fum.), 4.364Hm, 4.
11 4H m, 3.79 6H s, 3.16 2Hm,
2.77 4H m, 2.54 2H m, 2.40 2H
m, 2.22 4Hm, 2.01 4Hm, 1.894
Hm, 1.714Hm, 0.936Ht.

[0052]

Example 5 O, O′-dipropionyl (1,3-propylene) bispyrocarpe
Over preparative (Y = propionyl, A = methylene) by the method described in Example 2 O, O'dihydrogen (1,3-propylene) bispilocarpate (500 mg; 1.02 mmol) (see Example 1) Propionyl chloride (751mg; 8.12m
mol) and the title bispirocarpate was obtained. The yield was 467 mg (0.77 mmol). pK _a = 5.95 HR-MS- spectrum: m / e (relative intensity): 604
[M ^+. ] (6%), 517 (7%), 411 (13
%), 410 (57%), 396 (16%), 395
(71%), 339 (36%), 209 (15%), 2
08 (6%), 195 (48%), 121 (20%),
96 (56%), 95 (100%). HR-MS: Molecular weight = 604.3491210 (measured value) 604.3472151 (calculated value) NMR: δ 7.56 2H bs, 6.76 2H b
s, 4.18 4H m, 4.08 4H m, 3.62
6H s, 2.69 4H m, 2.49 2H m, 2.
33 2H m, 2.31 4H q, 2.01 4H q
v, 1.68 4H m, 1.10 6H t, 0.91
6H t.

[0053]

Example 6 O, O′-dicyclopropylcarbonyl (1,3-propylene) bi
Spirocarpet (Y = cyclopropylcarbonyl, A = methylene) By the method described in Example 2, O, O′-dihydrogen (1,3-propylene) bispyrocarpate (422 mg; 0.86 mmo)
l) (see Example 1) and cyclopropylcarbonyl chloride (719 mg; 6.88 mmol) gave the title bispyrocarpate. Yield 297 mg (0.47 mm
ol). pK _a = 6.05 HR-MS- spectrum: m / e (relative intensity): 629
(4%), 628 [M ^+. ] (7%), 423 (19
%), 422 (76%), 408 (16%), 407
(73%), 352 (10%), 351 (40%), 3
35 (9%), 334 (9%), 209 (19%), 2
08 (11%), 207 (73%), 121 (32
%), 96 (55%), 95 (100%). HR-MS: molecular weight = 628.3428340 (measured value) 628.3472151 (calculated value) NMR:? 7.51 2Hbs, 6.73 2Hb.
s, 4.19 4H m, 4.07 4H m, 3.61
6H s, 2.68 4H m, 2.49 2H m, 2.
33 2H q, 2.02 2H qv, 1.68 4H
m, 0.91 6H t, 0.90 8H m.

[0054]

Example 7 O, O′-dicyclobutylcarbonyl (1,3-propylene) bis
Pyrocarbate (Y = cyclobutylcarbonyl, A = methylene, fumarate) By the method described in Example 2, O, O′-dihydrogen (1,
3-Propylene) bispyrocarbate (705 mg; 1.
43 mmol) (see Example 1) and cyclobutylcarbonyl chloride (1357 mg; 11.45 mmol) gave the title bispyrocarpate. The fumarate salt was crystallized according to Example 4. The yield is 960 mg (0.9
6 mmol). Melting point = 33 to 35 ° C. (fum.) HR-MS-spectrum: m / e (relative intensity): 657
(4%), 656 [M ^+. ] (6%), 543 (9%),
437 (23%), 436 (100%), 422 (22
%), 421 (86%), 366 (10%), 365
(46%), 222 (9%), 221 (58%), 20
9 (14%), 121 (15%), 96 (68%), 9
5 (78%). HR-MS: molecular weight = 656.3743290 (measured value) 656.3785152 (calculated value) NMR: δ 8.50 2H bs, 7.23 2H b
s, 6.73s (fum.), 4.21 4H m, 3.
80 6H s, 3.16 2H m, 2.78 4Hm,
2.53 2H m, 2.40 2H m, 2.22 4H
m, 2.03 2Hqv, 2.01 4H m, 1.89
4H m, 1.71 4H m, 0.92 6H t. Correspondingly, 0,0'-dihydrogen (2-hydroxy-1,
3-Propylene) bispyrocarbate (544 mg; 1.
07 mmol) (see Example 1) and propionyl chloride (785 mg; 8.49 mmol) can give 0.0,0'-dipropionyl (2-hydroxy-1,3-propylene) bispyrocarpate.

[0055]

Example 8 O, O′-dipivalyl (1,2-ethylene) bispyrocarpate (Y = pivalyl, A = direct bond, fumarate) By the method described in Example 2, O, O′-dihydrogen (1 , 2-Ethylene) bispyrocarpate (384 mg; 0.80 mmol) (see Example 1) and pivalyl chloride (772 mg; 6.41 m)
mol) and the title bispirocarpate was obtained. The fumarate salt was crystallized according to Example 4 with a 2-propanol / toluene / petroleum ether mixture. Yield 293
It was mg (0.29 mmol). Melting point = 61 to 65 ° C. (fum.) HR-MS-spectrum: m / e (relative intensity): 647
(5%), 646 [M ^+. ] (7%), 645 (3%),
631 (6%), 531 (6%), 425 (24%),
424 (91%), 410 (17%), 409 (67
%), 337 (9%), 293 (7%), 223 (34
%), 209 (9%), 163 (9%), 123 (10
%), 122 (5%), 121 (29%), 96 (84)
%), 95 (100%). HR-MS: molecular weight = 646.39318800 (measured value) 646.39416531 (calculated value) NMR:? 8.51 2Hbs, 7.23 2Hb
s, 6.73 s (fum.), 4.37 4H m,
4.10 4H m, 3.80 6H s, 2.78 4H
m, 2.55 2H m, 2.39 2H m, 1.71
4H m, 1.20 18H s, 0.93 6H t.

[0056]

Example 9 O, O′-dicyclopentylcarbonyl (1,3-propylene) bi
Spirocarpate (Y = cyclopentylcarbonyl, A = methylene, fumarate) By the method described in Example 2, O, O′-dihydrogen (1,3-propylene) bispyrocarpate (401 mg;
0.81 mmol) (see Example 1) and cyclopentylcarbonyl chloride (859 mg; 6.48 mmol) gave the title bispyrocarpate. Fumarate is 2-
Crystallized with a propanol / toluene / petroleum ether mixture. The yield was 501 mg (0.48 mmol). Melting point = This compound is hygroscopic HR-MS-Spectrum: m / e (relative intensity): 685
(4%), 684 [M ^+. ] (6%), 683 (2%),
451 (11%), 450 (48%), 436 (7
%), 435 (25%), 379 (15%), 235
(29%), 209 (12%), 163 (12%), 1
21 (31%), 96 (47%), 95 (10%). H
R-MS: Molecular weight = 684.4060820 (measured value) 684.40984154 (calculated value) NMR: δ 8.572HHs, 7.27 2Hb
s, 6.72s (fum.), 4.21 4H t, 4.
10 4H d, 3.81 6H s, 2.80 4Hm,
2.75 2H m, 2.54 2H m, 2.40 2H
m, 2.03 2Hqv, 1.88 4H m, 1.74
4H m, 1.71 4H m, 1.62 8H m, 0.
926H t.

[0057]

Example 10 O, O′-dicyclohexylcarbonyl (1,3-propylene) bi
Spirocarpet (Y = cyclohexylcarbonyl, A = methylene, fumarate) By the method disclosed in Example 2, O, O′-dihydrogen (1,3-propylene) bisspirocarpate (424 mg;
0.86 mmol) (see Example 1) and cyclohexylcarbonyl chloride (1009 mg; 6.88 mmol) gave the title bispyrocarpate. Fumarate is
Crystallized with 2-propanol / toluene / petroleum ether mixture. The yield was 722 mg (0.68 mmol). Melting point = 50 to 53 ° C. HR-MS-Spectrum: m / e (relative intensity): 713
(2%), 712 [M ^+. ] (6%), 711 (3%),
465 (16%), 464 (54%), 450 (9
%), 449 (31%), 394 (5%), 393 (1
8%), 250 (6%), 249 (33%), 209
(12%), 163 (12%), 121 (32%), 9
6 (56%), 95 (100%). HR-MS: molecular weight = 712.4428410 (measured value) 712.4441156 (calculated value) NMR: δ 8.52 2H bs, 7.26 2H b
s, 6.71 s (fum.), 4.20 4H t,
4.09 4H d, 3.816 Hs, 2.79 4H
m, 2.54 2H m, 2.39 2H m, 2.32
2H m, 2.03 4Hqv, 1.87 4H m, 1.
74 4H m, 1.72 4H m, 1.70 4H m,
1.38 8H m, 0.92 6H t.

[0058]

Example 11 O, O′-succinyl (diethyl) bispyrocarpate (R = ethyl, B = direct bond) A solution of 9.80 mmol (2432 mg) of pilocarpine acid sodium salt in 60 ml of dimethylsulfoxide was added to 9 .80 mmol (10
68 mg) of ethyl bromide was added dropwise over about 1 hour. The solution was stirred at room temperature for 48-72 hours, 100 m
Pour into 1 liter of distilled water. The mixture was extracted with ethyl acetate in two portions of 150 ml each. The ethyl acetate extracts were combined, washed with 150 ml of distilled water, washed with 150 ml of 2% sodium bicarbonate solution, and further washed with 150 ml of distilled water. The ethyl acetate extract was dried over calcium sulfate for 30 minutes, the ethyl acetate was evaporated under reduced pressure, and the resulting bispyrocarpic acid ethyl ester was crystallized from a chloroform / petroleum ether mixture to give 448 mg of the title ester compound ( 1.76 mmol) was obtained. Mp _{= 104~107 ° C pK a = 6.60} HR-MS- spectrum: m / e (relative intensity): 254
[M ⁺ ] (17%), 236 (18%), 223 (11
%), 209 (24%), 207 (11%), 163
(14%), 139 (46%), 121 (31%), 9
6 (53%), 95 (100%). HR-MS: Molecular weight = 254.1620180 (measured value) = 254.1630428 (calculated value) NMR: δ 7.49 1H bs, 6.74 1H b
s, 4.13 2H m, 3.61 3Hs, 3.55
2H m, 2.73 1H m, 2.53 1H m, 2.
50 1H m, 2.03 2H m, 1.68 2H m,
1.27 3H t, 0.90 3H t. 98 mg (0.63 m) in a mixed solution containing 401 mg pilocarpic acid ethyl ester and 873 mg (6.32 mmol) calcium carbonate in toluene (50 ml).
(mol) succinyl chloride was added dropwise over about 24 hours. The resulting solution was stirred at room temperature for about 48 hours. A 2% sodium bicarbonate solution (50 m
1) was added and the resulting mixture was stirred at room temperature for 3 hours.
The resulting layers were separated and the toluene phase was washed twice with distilled water (2 x
(100 ml), washed with calcium sulfate for 30 minutes and evaporated under reduced pressure to give 192 mg (0.32 mmo) of O, O'-succinyl (diethyl) bispyrocarpate.
l) was obtained. HR-MS-spectrum: m / e (relative intensity): 590
[M ⁺ ] (9%), 545 (12%), 476 (24
%), 475 (90%), 424 (13%), 409
(7%), 237 (9%), 223 (12%), 209
(7%), 163 (9%), 121 (100%), 96
(39%), 95 (82%). HR-MS: Molecular weight = 590.3260650 (measured value) 590.3315650 (calculated value) NMR: δ 7.50 2H bs, 6.75 2H b
s, 4.16 4H m, 4.12 4H m, 3.61
6H s, 2.68 4H m, 2.62 4H m, 2.
46 2H m, 2.29 2H m, 1.67 4H m,
1.28 6Ht, 0.91 6Ht. The sodium salt of pilocarpic acid, which is the starting material, can be obtained according to the method described in Example 1.

[0059]

Example 12 O, O′-Glutaryl (diethyl) bispyrocarpate (R = ethyl, B = methylene) The ethyl ester of pilocarpine (589 mg; 2.32 by the method described in Example 11).
mmol) and glutaryl chloride (157 mg; 0.93 m)
mol) and the title bispirocarpate was obtained. The yield was 152 mg (0.25 mmol). HR-MS-spectrum: m / e (relative intensity): 604
[M ⁺ ] (7%), 559 (11%), 490 (21
%), 489 (75%), 237 (9%), 223 (6
%), 209 (8%), 163 (10%), 123 (6)
%), 122 (10%), 121 (100%), 96
(26%), 95 (61%). HR-MS: Molecular weight = 604.3478240 (measured value) 604.3472151 (calculated value) NMR: δ 7.53 2H bs, 6.74 2H b
s, 4.13 4H m, 4.08 4H m, 3.61
6H s, 2.67 4H m, 2.45 2H m, 2.
39 4H m, 2.30 2H m, 1.892 2H m,
1.67 4H m, 1.26 6H t, 0.90 6H
t.

[0060]

Example 13 O, O′-Adipoyl (diethyl) bispyrocarpate ( R = ethyl, B = ethylene) Ethyl ester of pilocarpine (609 mg; 2.4) by the method described in Example 11.
0 mmol) and adipoyl chloride (176 mg; 0.96
to give the title bispyrocarpate. The yield was 259 mg (0.42 mmol). HR-MS = spectrum: m / e (relative intensity): 618
[M ⁺ ] (10%), 573 (9%), 504 (20
%), 503 (67%), 489 (10%), 368
(10%), 365 (17%), 295 (9%), 24
9 (29%), 237 (11%), 236 (24%),
223 (15%), 209 (18%), 207 (20
%), 163 (21%), 123 (9%), 122 (1
2%), 121 (100%), 96 (51%). HR-MS: molecular weight = 618.36225030 (measured value) 618.3628652 (calculated value) NMR: δ 7.53 2H bs, 6.73 2H b
s, 4.13 4H m, 4.08 4H m, 3.61
6H s, 2.67 4H m, 2.472 2H m, 2.
34 4H m, 2.30 2H m, 1.68 4H m,
1.63 4H m, 1.26 6H t, 0.90 6H
t.

Continued Front Page (72) Inventor Suhonen, Pekka Finland, Sev -70200 Kuopio, Letokeryentier 9 Day 33 (72) Inventor Ulutti, Alt Finland, Sev -70420 Kuopio, Kerokuya 11 Be 10 (72) Inventor Hanhi Jelvi, Hannu Finland, SEF-20600 Turku, Paloker Jenkatu 1 Aes. 2 (72) Inventor Poyala, Esco Finland, Sef-33720 Tampere, Totorinkatu 7 Be 6

Claims (9)

[Claims] 1. The following general formula 1 A bispyrocarpic acid ester derivative represented by or an acid addition salt thereof. In Formula 1, A) Y is -C (= O) -R (R is an alkyl group having 1 to 4 carbon atoms,
A cycloalkyl group having 3 to 6 carbon atoms, or a phenyl group), and W is A group represented by [Y is as defined above, A is a direct bond, or a hydroxyl group, a group represented by YO- (Y is as defined above) and one or two Is a methylene group substituted or unsubstituted with a group selected from the group consisting of methyl groups, or B) W is an alkyl group having 1 to 4 carbon atoms or 3 to 6 carbon atoms.
Is a cycloalkyl group of Y and Y is (B is a direct bond, or is a methylene group or an ethylene group, and R ′ has the same meaning as W above). 2. The bis according to claim 1, wherein in the case of A), R is a cycloalkyl group having 3 to 6 carbon atoms, or a straight-chain or branched propyl or butyl. Pilocarpine derivative. 3. In the case of B), R ′ is a cycloalkyl group having 3 to 6 carbon atoms, or a straight chain or branched propyl or butyl group. Bispirocarpine derivative. 4. The case of A), wherein A is a direct bond, or is an unsubstituted methylene group or a methylene group substituted with a hydroxyl group. A bispirocarpine derivative of. 5. The bispirocarpine derivative according to claim 1 or 3, wherein in the case of B), B is a direct bond or an unsubstituted methylene group. 6. In the case of A), O, O'-dicyclopropylcarbonyl (1,2-ethylene) bispyrocarbate, O, O'-dicyclobutylcarbonyl (1,2-ethylene)
Bispyrocarbate, O, O'-dicyclopropylcarbonyl (1,3-propylene) bispyrocarbate, O, O'-dicyclobutylcarbonyl (1,3-propylene) bispyrocarbate, O, O '-Dicyclopropylcarbonyl (2-methyl-
1,3-propylene) bispyrocarbate, O, O'-dicyclopropylcarbonyl (2,2-dimethyl-1,3-propylene)
Bispyrocarpet, O, O'-dipropionyl (1,3-propylene) bispyrocarpate, O, O'-dicyclopropylcarbonyl (2-hydroxy-1,3-propylene) bispyrocarpate, O , O'-Dicyclopropylcarbonyl (2-cyclopropylcarbonyloxy-1,3-propylene) bispyrocarpate, O, O'-dipivalyl (1,2-ethylene) bispyrocarpate, O, O'- Dipivalyl (1,3-propylene) bispyrocarbate, O, O'-di (1-methylcyclopropylcarbonyl) (1,2-ethylene) bispyrocarbate, O, O'-dicyclopentylcarbonyl (1, 2-ethylene) bispyrocarbate, O, O'-dipropionyl (1,2-
Ethylene) bispyrocarpate, O, O'-diisobutyryl (1,2-ethylene) bispyrocarpate, O, O'-dipropionyl (2-hydroxy-1,3-propylene) bispyrocarpate, O, O'-dicyclohexylcarbonyl (1,2-ethylene) bispyrocarpate, O, O'-dicyclopentylcarbonyl (1,3-propylene) bispyrocarpate, O, O '
-Dicyclohexylcarbonyl (1,3-propylene) bispyrocarbate, O, O'-dibenzoyl (1,2-ethylene)
Bispirocarbate or O, O'-dibenzoyl (1,2-
The bispyrocarpine derivative according to claim 1, which is propylene) bispyrocarpate. 7. In the case of B), O, O'-succinyl (diethyl) bispyrocarpate, O, O'-succinyl (diisopropyl) bispyrocarpate, O, O'-succinyl (di-t- Butyl) bispyrocarpate, O, O'-succinyl (dicyclopropyl) bispyrocarpate, O, O'-
Succinyl (dicyclobutyl) bispyrocarbate, O,
O'-glutaryl (diisopropyl) bispyrocarpate, O, O'-glutaryl (di-t-butyl) bispyrocarpate, O, O'-glutaryl (dicyclopropyl) bispyrocarpate, or O, O '-Glutaryl (dicyclobutyl)
The bispirocarpine derivative according to claim 1, which is bispirocarpate. 8. A method for producing the compound according to any one of claims 1 to 7, wherein a) a compound represented by the formula: [Y is -C (= O) -R (R is an alkyl group having 1 to 4 carbon atoms, a cycloalkyl group having 3 to 6 carbon atoms, or a phenyl group), and A is a direct bond. Or, hydroxyl group, Y
A group represented by —O— (Y is as defined above) and a methylene group substituted or unsubstituted by a group selected from the group consisting of one or two methyl groups] The following general formula 5 Reacting a compound of formula (A is as defined above) with an acid of formula RCO ₂ H or a functional derivative thereof; or b) a compound of formula 6 (B is a direct bond, or is a methylene group or an ethylene group, and R'is an alkyl group having 1 to 4 carbon atoms or a cycloalkyl group having 3 to 6 carbon atoms). A compound represented by the general formula: (Where R ′ is as defined above) is represented by the general formula 8 (B is as defined above) is reacted with a dicarboxylic acid or a divalent acid derivative thereof, and
If desired, a process for the production, which comprises converting the compound obtained by the reaction into its acid addition salt. 9. A pharmaceutical composition comprising the compound according to any one of claims 1 to 7 as an active ingredient, and a pharmaceutically acceptable excipient.