JPS604815B2 - proline derivative - Google Patents

Abstract

The compounds of formula I are prepared by N-acylation of a compound of formula II' by means of an acid of formula III'. In these formulae, R is a hydroxyl, NH2 or alkoxy group, R1 and R4 are each H or an alkyl, phenyl or phenylalkyl group, R2 is H or an organic substituent, R3 is H or a hydroxyl or alkyl group, m is 1, 2 or 3 and n is 0, 1 or 2. The acid of formula III' can be replaced by a functional derivative of the latter or, when n is 1, by an acrylic acid of formula R4CH=C(R1)-COOH. In the latter case, after the acylation, a thiol or thioacid R2SH is added to the double bond of the acryloyl residue. The compounds of formula I are therapeutically active in the treatment of blood hypertension (high blood pressure). <IMAGE>

Description

[Detailed description of the invention]

The present invention relates to proline derivatives, and more particularly to novel proline derivatives and salts thereof having pharmacological activity. The novel proline derivatives of the present invention include compounds represented by the following general formula and salts thereof. The compounds of the present invention are useful as angiotensin converting enzyme inhibitors and can be used as antihypertensive agents. If the compound of the present invention is further clarified, it can be represented by the following formula. The symbols in the above formula have the following meanings. R and R4 each represent hydrogen or lower alkyl. R5 represents hydrogen or a group represented by the formula: n is 0, 1 or 2. The * mark in the formula represents an asymmetric carbon atom. However, the carbon atoms carrying R and R4 substituents are asymmetric when each substituent is a group other than hydrogen.
Among the compounds of the present invention [1], particularly preferred compounds have the formula: (
R,, R2 and n have the same meanings as above. ). Among the compounds m in the above range, the following compounds are particularly preferred embodiments of the compounds of the present invention [
1], the preference of which increases in the order of description (a to g). 'a}n is 1, 'b-R2 is hydrogen,

[c) R, is hydrogen or methyl, 'd] R, and R2 are each hydrogen, n
is 0, {e) R and R2 are each hydrogen, n is 1,
{f]R, is methyl, R2 is hydrogen, n is 1, (g) R2
is a group represented by the formula: (in which R is hydrogen or lower alkyl (especially methyl), and n is 0 to 2 (especially 1)). Naturally, compounds obtained by combining the above-mentioned preferred groups are also included in the preferred compounds. Stereoisomers in which the proline nucleus is in the L-form are particularly preferred compounds. Lower alkyl, represented by the various symbols, includes straight chain or branched hydrocarbon radicals from methyl to heptyl, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl, bentyl, isobentyl, and the like. The above groups are preferably C, -C4 groups (particularly C, -C2 groups). The compound [1] of the present invention (and its preferable sub-concept compounds) can be produced by the following method. Compounds of the present invention in which R2 is generally hydrogen [1]
is the formula: [wherein R represents hydroxy or lower alkoxy]. ] A compound represented by the formula: [wherein R,, R4 and n have the same meanings as above. ), and when R is lower alkoxy, it can be obtained by eliminating this. However, by acylation with an acid [machi], not only can the present compound [1] be obtained directly, but also the present compound [1] can be obtained via an intermediate. That is, the compound [m] is represented by the formula {a}: [wherein x represents halogen (preferably bromo, chloro or bromo). R,, R4 and n are the same as above. ] -haloalkanoic acid, or {b} formula: [wherein R,,R4
and n has the same meaning as above. ] tosyloxyalkanoic acid (compound [V] where x is tosyloxy), or 'c-formula: [wherein R
, and R4 have the same meanings as above. ], and the acylated product is then acylated with a substituted acrylic acid of the formula: R
5CO-SH [Skin] [In the formula, R5 represents lower alkyl, phenyl or phenyl lower alkyl. The intermediate of the compound [1] of the present invention can be obtained by substitution or addition with a thioic acid anion represented by the formula [1]. The compound of the present invention [1] or its ester, in which R2 is hydrogen, is obtained by converting the starting material [m] to the formula: [wherein n' represents 1 or 2]. R and R4 have the same meanings as above. It can be obtained by acylation with a thiolactone shown in ]. Alternatively, the starting material [m] can be of the formula: where Y may be R5CO or a protecting group (eg or other sulfur side protecting group). R, R, , R4, R5 and n have the same meanings as above. An intermediate can be obtained by acylation with a mercaptoalkanoic acid represented by ]. A compound in which Y is a protecting group as described above is a starting material for producing the target compound [1] in which R2 is hydrogen or its ester, and the compound [machi] is acylated with such starting material [K]. An intermediate of compound [1] in which Y is protected is obtained by a conventional method of treating this with hot trifluoroacetic acid, cold trifluoromethanesulfonic acid, mercuric acetate, sodium in liquid ammonia, zinc and hydrochloric acid, etc. R by deprotecting group reaction
The compound of the present invention [1] in which 2 is hydrogen or its ester can be obtained (for the removal treatment of the protecting group, refer to Methoden der Organitsien Chemie).
dender, OrganishenChemie (
(See The First Supper Part 1, page 736 and et seq.). When acid [W] or [Phantom] is used as the acylating agent for starting material [m], a coupling reagent such as dicyclohexylcarbodiimide or the like is used, or a mixed anhydride, symmetric anhydride, acid By forming a halide, acid ester or activating the acid using Woodward's reagent K, N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline, etc. Acylation treatment can be carried out effectively.
henChemie (Horien-Weyl)) No. XV
(See Volume Partro, page 1 and the following). Starting materials [m] include proline and its lower alkyl esters. The acylation treatment of the starting material [m] will be detailed below. A preferred method for producing an intermediate of compound [1] in which Y is R5-CO- is to combine the starting material [m] (an acid or its esters) with the formula: [wherein X is a halogen (preferably chloro or Bromo). R,, R4 and n have the same meanings as above. ] is subjected to a coupling treatment. This reaction treatment involves forming a mixed acid anhydride, symmetrical anhydride, acid halide, or active ester of acid [V] before reacting acid [V] with acid [m], or using Woodward reagent K.
, EEDQ (N-ethoxycarbonyl-2-ethoxy-
This can be carried out by activating the acid [V] using, for example, 1,2-dihydroxyquinoline, and then coupling the acid [V] with the compound [m]. By this operation, the formula: [wherein R, R, , R4, n and x have the same meanings as above. ] is obtained. This compound [X] has the formula: R5CO-SH [dish]
[In the formula, R5 has the same meaning as above. ] By subjecting the thioic acid to a substitution reaction with an anion, the formula: [wherein R, R,, R4, R5 and n have the same old meanings as above. ] An intermediate of the compound of the present invention can be obtained. By treating the above compound [phantom] with ammonium chloride, it was prepared by the formula: [wherein R, R, , R4 and n have the same meanings as above. ] or an ester thereof. When R is a lower alkoxy group, for example, when R is t-butoxy or t-amyloxy, by treating the ester form of the intermediate [Gen] or [Kari] of the compound of the present invention with trifluoroacetic acid and anisole. The corresponding free acid can be obtained by eliminating the lower alkoxy group. When the above alkoxy is present, the corresponding acid product can also be obtained by alkaline hydrolysis. The various manufacturing methods for obtaining the compounds of the present invention explained above will be explained in more detail as follows. Formula: [wherein R and R4 have the same meanings as above. ] The acrylic acid starting material represented by is transformed into its acid halide form, and then this is reacted with compound [m] to obtain the formula: where R, R, and R4 have the same meanings as above. ], and convert it into the formula: R5CO-
An intermediate of the compound of the present invention can be obtained by subjecting it to an addition reaction with a thioic acid represented by SH. Further, as described above, the formula: [wherein R,, R4 and n have the same meanings as above. The intermediate [M] of the compound of the present invention is obtained by acylating the starting material [m] using tosyloxyalkanoic acid represented by ] as an acylating agent, and subjecting this acylated product to the above-mentioned substitution reaction. be able to. Also, by reacting acrylic acid and thioic acid, the formula: [where n,
R,, R4 and R5 have the same meanings as above. ] is prepared, this is treated with, for example, thionyl chloride to convert the acid halide, this is coupled with the starting material [m], and further treated as described above to obtain the present invention. An intermediate (red) of the invention compound can be obtained. The acid or ester starting material [m] is represented by the formula: [In the formula, R8 represents a protecting group (which may be the same protection as described above). n, R, and R4 have the same meanings as above. ] It can be acylated with a protected type compound of -mercaptoalkanoic acid represented by . In this case, the compound of the present invention [1] can be obtained by carrying out the conventional deprotection treatment as described above after the acylation treatment. Furthermore, a thiolactone type compound (for example, 8-propiothiolactone, Q-methyl-8-propiothiolactone, etc.) can be used as an acylating agent for the starting material [m]. A preferred method for producing the compound of the present invention will be explained in detail in Examples below. Representative examples of particularly preferred production methods for obtaining the compounds of the present invention are as follows. The acid or its ester starting material [Rainbow] is expressed by the formula: (where ×
each individually a halogen (preferably chloro or bromo), R, , R4 and n have the same meanings as above. Acylated with haloalkanoyl halide shown in ]. This reaction operation can be carried out in an alkaline medium (eg dilute alkali metal hydroxide solution, alkali metal bicarbonate solution or alkali metal carbon solution) at low temperature (eg about 0 to lyo). An intermediate can be obtained by subjecting this acylation reaction product to a substitution reaction with a thioic acid anion. This treatment is carried out in an alkaline medium (
This can be carried out in a conventional manner (preferably in an alkali metal carbonate solution). This reaction product was treated with ammonolysis (
Compounds of the invention in which R2 is hydrogen can be obtained by treatment with an alcoholic ammonia or concentrated ammonium hydroxide solution) or alkaline hydrolysis (eg, treatment with an aqueous metal hydroxide solution). Further, as a representative example of a particularly preferred method for producing the compound of the present invention, the ester starting material [m] (preferably t-butyl ester) is prepared with the formula: [
In the formula, R,, R4, R5 and n have the same meanings as above. ] The method of treatment with a thioalkanoic acid can be mentioned. This operation is carried out in an anhydrous medium such as dichloromethane, tetrahydrofuran, dioxane, etc. in the presence of dicyclohexylcarbodiimide, N'N'-carbonylbisimidazole, ethoxyacetylene, diphenylphosphoryl azide or similar coupling agent from about 0 to
This can be done with 1oo0. A compound in which R2 is hydrogen is obtained by subjecting this reaction product to ammonolysis treatment as described above. When R is a lower alkoxy group, it can be substituted with, for example, trifluoroacetic acid and anisole (
By treatment at room temperature), the lower alkoxy group can be removed and the corresponding free acid can be obtained. When ester form [m] (a compound in which R is lower alkoxy (especially t-butoxy)) is acylated with a thiolactone (e.g. 3-propiothiolactone, Q-methyl-B-propiothiolactone, etc.), this The reaction is carried out in anhydrous solvents such as tetrahydrofuran, dioxane, methylene chloride, etc.
This is achieved by processing at about 0°C to room temperature. By treating this ester product with anisole and trifluoroacetic acid as described above, the ester group can be removed. Rmi: [wherein R,, R4 and n are as defined above. The compound of the present invention which is a group represented by ] can be obtained by directly oxidizing a compound [fox] in which R2 is hydrogen with iodine. The compound [1] of the present invention has one or more asymmetric carbon atoms. When R and R4 are groups other than hydrogen, the carbon atoms bonding them are asymmetric. These carbon atoms have the formula [
1] Indicated by *. Thus, the compounds of the present invention exist as stereoisomers or racemates thereof. All of these are included within the scope of the compounds of the present invention. Racemic compounds or optical antipodes can be used as starting materials in the production method. When racemic starting materials are used in the process of the invention, the stereoisomers in the product can be separated by conventional chromatography or fractional crystallization techniques. In general, L regarding the carbon atom of amino acids
- The isomers constitute preferred isomeric forms of the compounds. Also preferred is the D-isomer with respect to the first carbon in the acyl side chain of the compound of the present invention (i.e., the carbon atom to which R is attached). The compound [1] of the present invention forms basic salts with various inorganic or organic bases, and these salts are also included within the scope of the compounds of the present invention. Such salts include, for example, ammonium salts, alkali metal salts (
Examples include sodium salts and potassium salts, and these are preferred. ), alkali metal salts (e.g. calcium salts, magnesium salts), salts with organic bases such as dicyclohexylamine salts, benzathine salts, N-methyl-D-glucamine salts, hydrabamine salts, with amino acids such as arginine or lysine. salts, etc. Physiologically acceptable non-toxic salts are preferred, but other salts are also useful; for example, in the case of the dicyclohexylamine salt, other salts may be used to isolate and purify the dicyclohexylamine salt, as explained in the Examples below. is useful. Such salts can be prepared by the conventional process of reacting the free acid form product with an equimolar amount or more of a suitable base that provides the desired cation in a solvent or medium that does not dissolve it, or in water, and dehydrating it by freeze-drying. can be manufactured. Next, H-type cation exchange resin (e.g. DOWEX 5
The free acid form of the compound can be obtained by neutralizing the above salt with an insoluble acid or an aqueous acid solution such as polystyrene sulfonic acid resin (e.g., polystyrene sulfonic acid resin) and extracting with an organic solvent (e.g., ethyl acetate, dichloromethane, etc.). Other corresponding salts can also be prepared from the free acid if desired. The detailed manufacturing method of the present invention will be described in detail in Examples below. The examples represent preferred embodiments,
This suggests that compounds of the present invention not described in the examples can be similarly produced. The compounds of the present invention can inhibit the conversion of decabeptide angiotensin 1 to angiotensin lo, and are therefore useful for reducing or rescuing hypertension associated with angiotensin. The action of the enzyme renin on angiotensinogen (pseudoglobulin in blood acid) is to produce angiotensin-1. Angiotensin 1 is converted into angiotensin by angiotensin converting enzyme (ACE). The latter are antihypertensive active substances that have been implicated as causative agents in various types of hypertension in mammals (eg rats, dogs). The compound of the present invention inhibits angiotensin-converting enzyme,
mediate the angiotensin (renin) → angiotensin 1 → angiotensin lo conversion system by reducing or preventing the production of . Therefore, by administering the compound of the present invention or one of its physiologically acceptable salts or a mixture thereof to a mammal affected by angiotensin-dependent hypertension, its hypertensive symptoms can be inhibited. As shown in the typical animal studies described by Engel et al., a suitable blood pressure lowering dose of the compounds of the present invention is about 0.1 to 100 pm/k9/day, preferably about 1 to 50 9/k/day. Daily allowance 1 based on k9/day
times, preferably in 2 to 4 divided doses (for animal studies Engel (S.L. En bag 1), Schafer (T
．． R Schaeffer), Waug (M, J. Waug)
h), B. Rubin: Proc. Soc.
．． Exp. Biol. Med. 143, page 483 (19
(See King 72). The preferred administration method of the present invention is oral administration, but parenteral administration such as subcutaneous, intramuscular, intravenous, and intraperitoneal administration may also be used. The compounds of the present invention can be used to achieve a blood pressure reducing effect by formulating them into compositions such as tablets, capsules, elixirs for oral administration, or sterile solutions or suspensions for parenteral administration. can do. For this purpose, the compound of the present invention [1
] or a physiologically acceptable salt thereof or a mixture thereof in a physiologically acceptable medium, carrier, excipient, in a unit dosage form compatible with pharmacologically acceptable practice;
It can be formulated with binders, preservatives, stabilizers, and flavoring agents. The amount of active compound in these compositions or medicaments should be such that a dosage within the ranges described above will be administered. The following are examples of adjuvants that can be incorporated into tablets, capsules, etc. Binders such as gum tragacanth, gum arabic, cornstarch, gelatin, excipients such as dicalcium phosphate, disintegrants such as cornstarch, potato starch, alginic acid, etc., lubricants such as stearic acid, sucrose, Sweeteners like lactose, saccharin, and flavoring agents like peppermint, oil of wintergreen, and cherry flavor. When the dosage unit form is a capsule, it can contain, in addition to the above materials, a liquid carrier such as a fatty oil. Various other materials can be used as coating agents to coat the dosage unit form or to modify its external shape. For example, tablets may be coated with shellac, sugar or both. syrups, elixirs, active compounds,
It may contain sucrose as a sweetening agent, methyl and propyl parabens as preservatives, and cherry flavor or orange flavor as a coloring agent and flavoring agent. Sterile injectable compositions include the active substance and a natural vegetable oil (such as sesame oil, coconut oil, peanut oil, cottonseed oil, etc.) or a synthetic resinous carrier (such as ethyl oleate) in a carrier such as sterile water for injection. They can be dissolved or suspended and formulated according to common pharmaceutical practice. If necessary, a buffer, a preservative, an antioxidant, etc. can be added. Next, specific methods for producing preferred compounds of the present invention will be described in detail with reference to Examples and Reference Examples. Reference Example 11 Method for producing -(2-benzoyltheoacetyl)-L-proline: 5.75 mL of -L-proline is dissolved in 50% of IN sodium hydroxide, and the solution is cooled in an ice-water solution. 26 sodium hydroxide and 5 chloroacetyl chloride
．． Add 65 ml of water and shake the mixture vigorously for 3 hours at room temperature.
50% water, 7.5% underground thiobenzoic acid and 4% potassium carbonate
．． After 8 hours of suspension and incubation at room temperature for 1 hour, the mixture is acidified and extracted with ethyl acetate. The ethyl acetate layer is washed with water, dried over magnesium sulfate, and then concentrated to dryness under reduced pressure. 14.6 ml of the residue was dissolved in 150 M ethyl acetate,
Add dichlorohexylamine 11. The produced crystals were separated from the furnace and recrystallized from ethyl acetate to obtain product 5.
．． Get 7 evenings. Melting point: 151-15 or 0. This crystalline salt was mixed with 10Q of 5% potassium hydrogen sulfate aqueous solution and 30Q of ethyl acetate.
Dissolve in the ground mixture. The organic layer was washed once with water, dried over magnesium sulfate, and concentrated to dryness under reduced pressure to obtain the acid product 3.
．． I got 45 evenings. Example 1 Production of 1-(2-mercaptoacetyl)-L-proline: - 3.4 g of 1-(2-penzoylthioacetyl)-L-proline obtained in Reference Example 1 was mixed with 10.5 g of water [ and concentrated ammonia dissolved in a mixture of 6.4'. After 1 hour, the mixture is diluted with water and filtered. The filtrate is extracted with ethyl acetate, acidified with concentrated acid, saturated with sodium chloride and extracted twice with ethyl acetate. The ethyl acetate extract is washed with saturated sodium chloride and concentrated to dryness to yield 1.5 liters of product. This was crystallized from ethyl acetate to obtain 1-(2-mercaptoacetyl)-L-proline. Melting point 133-135°C. Reference Example 21-(2-benzoylthiopropanoyl)-L-ph. . Preparation of phosphorus: Dissolve 5.75 liters of L-proline in 50M IN aqueous sodium hydroxide solution and cool the solution in an ice bath. is sodium hydroxide 25' and 2-furomop. 8.57 g of pionyl chloride are added in that order, the mixture is removed from the ice and concentrated for 1 hour at room temperature. A mixture of 7.5 parts of thiobenzoic acid, 4.8 parts of potassium carbonate and 50 parts of water is added and the mixture is allowed to stand at room temperature overnight. After acidifying with concentrated hydrochloric acid, the aqueous solution is extracted with ethyl acetate, and the organic layer is washed with water, dried, and concentrated to dryness. The residue 14.7 mL was poured onto a silica gel 440 column, benzene-acetic acid (7:1).
The mixture is subjected to chromatography as an eluent. The fractions containing the desired material were combined and dried, and the residue was converted to the dicyclohexylamine salt by precipitating twice with ether-hexane in ether-hexane, yielding the product 9.4%. obtain. Melting point (14 is 0), 148-156°C. This dicyclohexylamine salt was treated in the same manner as in Reference Example 1 to obtain 5.7 g of the free acid. Example 21 Preparation of 1-(2-mercaptopropanoyl)-L-proline: 1-(2-penzoylthioyl) obtained in Reference Example 2 was added to a mixture of 12 parts of water and 9 parts of concentrated ammonium oxide. Dissolve 5.7 liters of L-proline (propanoyl). After 1 hour, the mixture is diluted with 10 parts of water and filtered. Extract the furnace liquid twice with ethyl acetate and add it to 1/3 of the original volume.
Concentrate, acidify with concentrated hydrochloric acid, and extract with ethyl acetate.
The organic layer is washed with saturated sodium chloride, dried, and then concentrated to dryness under reduced pressure. The residue was crystallized from ethyl acetate-hexane to obtain 1-(2-mercaptopropanoyl)-L-proline. Melting point (10500) 116-1200
0. Reference Example 31 Preparation of -(3-benzoylthiopropanoyl)-L-proline: 5.75 kg of -L-proline is dissolved in 50 kg of IN sodium hydroxide, and the solution is cooled in an ice bath. 8.5 g of 3-bromopropionyl chloride and 27 g of sodium hydroxide are added and the mixture is heated in an ice bath for 10 minutes and at room temperature for 3 hours. A suspension of 7.5 parts of thiobenzoic acid, 4.5 parts of potassium carbonate and 50 parts of water is added and the mixture is allowed to stand at room temperature for 1 hour. After acidifying with concentrated hydrochloric acid, the aqueous layer is extracted twice with ethyl acetate. The organic layer was dried over magnesium sulfate and then concentrated to dryness under reduced pressure to obtain 7.1 g of 1-(3-besozoylthiopropanoyl)-L-broline. Melting point 101-102
00 (ethyl acetate-hexane). Reference example Production of 4L-proline/t-butyl ester: -L-proline 230
Evening water 1000' and state sodium hydroxide 400M
dissolve in a mixture of Cool the solution in an ice bath, and divide 460' of sodium hydroxide and 340' of penzyloxycarbonyl chloride into 5 equal parts while shaking vigorously. Add over time. After lighting for 1 hour at room temperature, the mixture was extracted twice with ether and acidified with concentrated hydrochloric acid. The precipitate is dried in an oven. Yield 442 evenings. Melting point 78-80qo. 480 g of the obtained pendyloxycarbonyl-L-proline is dissolved in a mixture of 300 g of dichloromethane, 800 g of liquid isobutylene and 7.2 g of concentrated sulfuric acid. Shake the solution in a pressure bottle for 7 hours. After releasing the pressure and evaporating the isobutylene, the solution was washed with 5% sodium carbonate and water, dried over magnesium sulfate, and concentrated to dryness under reduced pressure to give penzyloxycarbonyl-L-proline-t-butyl ester 205. . Penzyloxycarbonyl-
Dissolve 250 grams of L-proline t-butyl ester in 1,200 grams of absolute ethanol and dissolve at 10% until no trace of carbon dioxide is observed in the hydrogen gas at the outlet (2 hours).
Hydrogenate with palladium/carbon (10 min) at normal pressure. The catalyst was separated into a furnace, and the furnace liquid was concentrated under reduced pressure (3 hours).
The residue was distilled under reduced pressure to obtain L-proline t-phthyl ester. Boiling point (1 willow) 50-5ro. Reference example 5
1. Method for producing 1-(3-acetylthiopropanoyl)-L-proline/t-butyl ester: - Dissolve 5.13 parts of the L-proline/t-butyl ester obtained in Reference Example 4 in 40 parts of dichloromethane, and pour the solution over ice and water. Cool inside. To this is added 6.18 molar dicyclohexylcarbodiimide in a 20M solution in dichloromethane, followed immediately by 4.45 molar 3-acetylthiopropionic acid. After heating in an ice water bath for 15 minutes and at room temperature for 16 hours, the precipitate was filtered out, and the furnace liquid was concentrated and dry distilled under reduced pressure. The residue is dissolved in ethyl acetate and washed neutral. After drying the organic layer over magnesium sulfate, it was concentrated to dryness under reduced pressure to obtain 1-(3-acetylthiopropanoyl)-L-proline/t-butylester9.
I got 8 evenings. Reference Example 6 Production of 1-(3-acetylthiopropanoyl)-L-proline: -1-(3-acetylthiopropanoyl)-L-proline/t-butyl ester obtained in Reference Example 5 4.7
The mixture is dissolved in a mixture of 34' of anisole and 68' of trifluoroacetic acid and the mixture is kept at room temperature for 1 hour. The solvent is evaporated under reduced pressure and the residue is precipitated several times from ether-hexane. 3.5 parts of this residue was dissolved in 25 parts of acetonitrile, and 2.8 parts of dicyclohexylamine was added. The crystalline salt is removed from the oven and recrystallized from isopropanol. Yield 3.8 hours, melting point 176-1770. This salt was converted into an acid in the same manner as in Reference Example 1, and 1-(3-
1.25 mL of (acetylthiopropanol)-L-proline was obtained. Melting point 89-90oo (ethyl acetate-hexane)
. Reference example 7 1-(3-captopropanoyl)-L-proline.
Production of t-butyl ester: 1 L-proline/t-butyl ester 3.42 hours of dry tetrahydrofuran solution was cooled in an ice bath, and propiothiolactone 1 was added to it.
．． Add 76 evenings. After being kept in an ice bath for 5 minutes and at room temperature for 3 hours, the reaction mixture is diluted with 200% ethyl acetate and washed with 5% potassium hydrogen sulfate and water. Dry the organic layer with magnesium sulfate,
The mixture was concentrated to dryness under reduced pressure, and the residue was crystallized from ether-hexane to obtain 3.7 g of 1-(3-mercaptopropanoyl)-L-proline/t-butyl ester. Melting point 5
7-5 is C. Example 3 Production of 1-(3-methylcaptopropanoyl)-L-proline: -Production method A:- 4.9 hours of 1-(3-penzoylthiopropanoyl)-L-proline obtained in Reference Example 3 Dissolve in a mixture of 8 parts water and 5.6 parts concentrated ammonium hydroxide and keep under argon atmosphere for 1 hour. The reaction mixture is diluted with water, filtered and the filtrate is extracted with ethyl acetate. The aqueous layer is acidified with concentrated hydrochloric acid, saturated with sodium chloride, and then extracted with ethyl acetate. The organic layer is washed with saturated sodium chloride, dried over magnesium sulfate, and concentrated to dryness under reduced pressure. The residue was crystallized from ethyl acetate-hexane to give 2.5 liters of 1-(3-methylcaptopropanoyl)-L-proline. Melting point 68-70qo. Manufacturing method B
5. 1-(3-acetylthiopropanoyl)-L-proline obtained in Reference Example 6 was mixed with 0.8 g of methanol/salt. Dissolve the speckled ammonia in 5' of ammonia and keep the solution at room temperature under an argon atmosphere. After 2 hours, the solvent was removed under reduced pressure and the residue was dissolved in water.
Ion exchange column (Dawex 50 (
Analytical grade)) and eluted with water. The fractions showing a theol positive reaction were combined and concentrated to dryness to yield 0.6% of the product. This was crystallized from ethyl acetate-hexane in the same manner as in Production Method A above to obtain 1-(3-mercaptopropanol)-L-proline. Production method C: 1-(3-mercaptopropanoyl)- obtained in Reference Example 7
L-proline t-butyl ester
Dissolve in a mixture of 20' of 100% trifluoroacetic acid and 45' of trifluoroacetic acid. After one hour at room temperature under an argon atmosphere, the reaction mixture is concentrated to dryness under reduced pressure and the residue is precipitated several times from ethyl acetate-hexane. 1.9% of the remaining liquid was dissolved in ethyl acetate 30%
of dicyclohexylamine and add 1.85% of dicyclohexylamine. The product was collected and recrystallized from isopropanol to obtain two crystalline salts. Melting point 187-188 qo. This salt was converted into an acid in the same manner as in Reference Example 1 to obtain product 1.3. This was crystallized from ethyl acetate in the same manner as in Production Method A above. Preparation of salts:- Sodium salt: 1-(3-Mercaptopropanol)-L-Proline 500 parts 9 is dissolved in a mixture of 2.5 parts of water and 2.5 parts of IN sodium hydroxide, and this solution is dissolved. The sodium salt was obtained by lyophilization. Magnesium salt: 1-1 (3-mercaptopropanol)
One L-proline 500 evening, magnesium oxide 49.5
9 parts of water and 10 parts of water are concentrated and slightly flooded until it becomes a complete solution. This was lyophilized to remove the solvent to obtain a magnesium salt. Calcium salt: 1-(3-captopropanol)
-2-Proline 500:9 was dissolved in a mixture of 91 parts sodium hydroxide and 10 parts water, and this solution was freeze-dried to obtain its calcium salt. Potassium salt: 1-(3-mercaptopropanoyl)-L
-Proline 500 was dissolved in a mixture of 9 parts potassium bicarbonate 246 parts and water 10 parts, and this was lyophilized to obtain a potassium salt. N-methyl-D-glucamine salt: 1-(3-captopropanoyl)-L-proline 500 female and N-methyl-
480 parts of D-glucamine was dissolved in 10 parts of water and lyophilized to obtain N-methyl-D-glucamine salt. Example 4 Preparation of 1-(3-captopropanoyl)-D-proline: - Using D-proline in place of L-proline in the treatment of Reference Example 3, the same treatment was carried out, and then this product was used in Example 3. 3 Process in the same manner as manufacturing method A to obtain 1-(3
-penzoylthiopropanoyl)-D-proline and 1-(3-mercaptopropanoyl)-D-proline (
Melting point 68-70oo) was obtained. Reference Example 8 Preparation of 3-acetylthio-2-methylpropanoic acid: A mixture of 50 ml of thioacetic acid and 40.7 ml of methacrylic acid is heated on a steam stove for 1 hour, and then kept at room temperature for 1 hour. After confirming the completion of the reaction of methacrylic acid by NMR spectroscopy, the reaction mixture was distilled under reduced pressure to obtain a boiling point of 128.5.
~13r0 (2.6 Yanagihiyu) crab fraction is separated and the desired 3
-Acetylthio-2-methylpropanoic acid (64%) was obtained.
Reference Example 9 Production of 3-penzoylthio-2-methylpropanoic acid:-
Thiobenzoic acid was used instead of thioacetic acid in the treatment of Reference Example 8, and the same treatment was performed to obtain 3-benzoylthio-2-methylpropanoic acid. Reference Example 10 Production of 3-phenylacetylthio-2-methyl-propanoic acid: - Using thiophenylacetic acid instead of thioacetic acid in the treatment of Reference Example 8, 3-phenylacetylthio-2-methyl was produced in the same manner as in Reference Example 8. Propanoic acid was obtained. Reference example 11 1-(3-acetylthio-2-methylpropanoyl)-
Production of L-proline t-butyl ester: -L-proline t-butyl ester 5.1 to dichloromethane 4
0' and cool the solution in an ice bath. To this was added a solution of 6.2 g of dicyclohexylcarbodiimide in dichloromethane for 15 days, and then immediately a solution of 4.9 g of 3-acetylthio-2-methylpropanoic acid obtained in Reference Example 8 in dichloromethane for 5 g. After incubating in an ice bath for 15 minutes and at room temperature for 1 hour, the precipitate was separated in the oven, and the solution was concentrated under reduced pressure in a dry column. The residue is dissolved in ethyl acetate and washed neutral. After drying the organic layer with magnesium sulfate,
It was concentrated to dryness under reduced pressure, and the residue was purified by column chromatography (silica gel-chloroform) to obtain 11(3-acetylthio-2-methylpropanoyl)-L-proline/t-butyl ester (7.9%). Reference example 12 1-(3-acetylthio-2-methylpropanol)-
Production of L-proline: 1. Production method A: - 1-(3-acetylthio-2-methylpropanoyl)-1 L-proline t-butyl ester (product of Reference Example 11) 7.8 ml was mixed with 55 ml of anisole and 1 ml of trifluoroacetic acid.
Dissolve in a 10' mixture. After one hour at room temperature, the solvent is removed under reduced pressure and the residue is precipitated several times from ether-hexane. Zancho 6.8
Dissolve the solution in 40ml of acetonitrile and add 4.5ml of dicyclohexylamine. The crystalline salt is boiled with 100 g of fresh acetonitrile, cooled to room temperature and filtered to give 3.8 g of the salt. Melting point (165) 187-18 is o
. This material is recrystallized from isopropanol. [Q]
D-670 (concentration in ethanol c: 1.4). This crystalline dicyclohexylamine salt is suspended in a mixture of 5% aqueous potassium sulfate and ethyl acetate. The organic layer was washed with water, concentrated to dryness, and the residue was crystallized from ethyl acetate-hexane.
-(3-acetylthio2-D-methylpropanoyl)
-L-proline was obtained. Melting point 83-8y0. [Q] Customer-
1620 (concentration in ethanol c=1.7). Preparation method B: 8.1 parts of 3-acetylthio-2-methylpropanoic acid and 7 parts of thionyl chloride are mixed, and this suspension is incubated at room temperature for 1 hour. The reaction mixture was concentrated and dried, and then distilled under reduced pressure (boiling point 8000
)do. 5.4 parts of the resulting 3-acetylthio-2-methylpropanoic acid chloride and 15 parts of sodium hydroxide are added to a solution of 3.45 parts of L-proline and 30 parts of IN sodium hydroxide (chilled in an ice-water bath). 3 at room temperature
After cooling for a period of time, the mixture was extracted with ether, the aqueous layer was acidified, and this was extracted with ethyl acetate. The organic layer was dried over magnesium sulfate and concentrated to give 1-(3-acetylthio-2-DL-methylprop/yl)-L-proline. Preparation C: - Dissolve 3.45 parts of L-proline in a mixture of 100 parts of water and 12 parts of sodium bicarbonate, cool the solution in an ice-water bath, and add 4 parts of methacryloyl chloride to it while stirring vigorously. Add 16 min. After the addition is complete, the mixture is kept at room temperature for 2 hours and extracted with ether. The aqueous layer is acidified with IN hydrochloric acid and extracted with ethyl acetate. The organic layer is concentrated to dryness under reduced pressure, the residue is mixed with 3.5 hours of thiol acetic acid, a few crystals of azobisisobutyronitrile are added and the mixture is heated on a steam stove for 2 hours. The reaction mixture was dissolved in benzene-acetic acid (75:25) and placed on a silica gel column eluting with the same solvent mixture.
-(3-acetylthio2-DL-methylpropanoyl)-L-proline was obtained. Reference example 13 1-(3-benzoylthio-2-methylpropanoyl)
-Production of L-proline/t-butylester: 1. Using 3-benzoylthio-2-methylpropanoic acid obtained in Reference Example 9 instead of 3-acetylthio-2-methylpropanoic acid in the treatment of Reference Example 11, the same treatment was carried out. te11 (3
-benzoylthio-2-methylpropanoyl)-1-L-broline t-butyl ester was obtained. Reference Example 14 Production of 1-(3-phenylacetylthio-2-methylpropanoyl)-L-proline t-butyl ester:-
In place of 3-acetylthio-2-methylpropanoic acid in the treatment of Reference Example 11, 3-phenylacetylthio-2-methylpropanoic acid obtained in Reference Example 10 was used and treated in the same manner as 1-(3-methylpropanoic acid). Phenylacetylthio-2-methylpropanoyl)-L-proline t-butyl ester was obtained. Reference example 15 1-(3-penzoylthio-2-methylpropanoyl)
-Production of L-proline: -Reference Example 1 Instead of 1-(3-acetylthio)-12-methylpropanoyl)-1-proline/t-butyl ester in the treatment of Neck Preparation Method A, the 1-( obtained in Reference Example 13) Using 3-benzoylthio-2-methylpropanoyl)-L-proline t-butyl ester, 1-(3-benzoylthio-2
-methylpropanoyl)-L-proline was obtained. Reference Example 16 Production of 1-(3-phenylacetylthio-2-methylpropanoyl-L-proline): -Reference Example 1 Production of 1-(3-acetylthio-2-methylpropanoyl)-L-proline in the treatment of Basic Production Method A. 1-(3-phenylacetylthio 2) obtained in Reference Example 14 instead of t-butyl ester
Example 5 11 (3-phenylacetylthio 2-methylpropanoyl 1 L-proline) was obtained by the same treatment using 11 (3-phenylacetylthio 2-methylpropanoyl 1 L-proline) and t-butyl ester. -2-D-methylpropanol)
-Production of L-proline: -Reference Examples 12, 15 and 16
Each of the products was treated as described below to give the title compound. 0.85 methanol and 5.0 methanol of the thioester product, respectively. Dissolve in ammonia and keep the solution at room temperature for 2 hours. The solvent was removed under reduced pressure, the residue was dissolved in water and applied to an ion exchange column (Dawex 50 (analytical grade)) on an H+ cycle.
eluted with water. Fractions giving a positive thiol reaction are combined and lyophilized. The residue was crystallized from ethyl acetate-hexane to give 1-(3-mercapto-2-D-methylpropanoyl-L-proline), melting point 1.
03-104qo, [Q]o-13r (in ethanol,
concentration c=2). Reference Example 17 Production of 1-(4-penzoylthiobutanoyl)-L-proline: -L-proline 2.88 mL of IN sodium hydroxide 25' solution was cooled in an ice bath, and sodium hydroxide was added to it. Add 12.5 ml of 4-chlorobutyl chloride and 3.5 ml of 4-chlorobutyl chloride. The mixture is allowed to stand for 3.5 hours at room temperature and to this is added a suspension of 3.75 parts of thiobenzoic acid, 2.4 parts of potassium carbonate and 25 parts of water. After stirring for 1 hour at room temperature, the reaction mixture is acidified with concentrated hydrochloric acid and extracted with ethyl acetate. The organic layer is dried over magnesium sulfate and concentrated to dryness under reduced pressure. The residue is chromatographed on a silica gel column using benzene-acetic acid (7:1) as eluent. The fractions containing the desired material are combined and concentrated to dryness to yield 1.35% of the product.
Dissolve a small amount of this material in ethyl acetate and adjust the pH of the solution to
Add dicyclohexylamine until 8-10 (on wet pH paper). The dicyclohexylamine salt crystallizes out immediately. Melting point 159-161oo. Example 61 Preparation of -(4-mercaptobutanoyl)-L-proline:
- 1.08 g of 1-(4-benzoylthiobutanoyl)-L-proline obtained in Reference Example 17 is dissolved in a mixture of 4 g of water and 2.7 g of concentrated ammonia. After heating at room temperature for 1 hour, the mixture is diluted with water, filtered and extracted with ethyl acetate, and the aqueous layer is concentrated under reduced pressure. The resulting 1-(4-mercaptobutanoyl)-L-proline ammonium salt was subjected to ion-exchange chromatography on a dimethylaminoethyl-cephadex (dextrin-crosslinked) column with ammonium bicarbonate gradient eluent. Purification is performed to obtain a pure product of 0.7%.
The ammonium salt is dissolved in 2 parts of water, treated on a Dowex 50 sulfonic acid resin (analytical grade, hydrogen form) column, and the resulting free acid is eluted with water. Fractions containing the desired substance (positive for sulfhydryl reagent and carboxyl reagent) were combined and lyophilized to obtain 1-(4-mercaptobutanoyl)-L-proline. This was further treated in the same manner as in Reference Example 17 to obtain its hexylammonium salt. Melting point 157-158°C. Reference Example Production of 184-furomo-2-methylbutanoic acid: Ethyl -4-furomo-2-methylbutanoate (see G. Jones and J. Wood: Tetrahedron Letters Vol. 21, 2961 (King 1968)) 1.04 minutes was dissolved in 50M dichloromethane and cooled to -10°C. While holding this in dichloromethane, boric tribromide I
Add 5' of M solution dropwise and further boil at 11oo○ for 1 hour, 25
Continue to pray for 2 hours at ℃. Terminate the reaction by carefully adding water. Each layer was separated, and the organic layer was washed with water, dried, and concentrated to dryness to obtain 4-furomo-2-methylbutanoic acid. Reference example 19 1-(4-penzoylthio-2-methylbutanoyl)-
Production of L-proline: - [a] 4-furomo-2-methylbutanoic acid obtained in Reference Example 18 and thionyl chloride were mixed and mixed at room temperature for 1 hour. The reaction mixture is concentrated to dryness and distilled under reduced pressure. 'b}L
-Proline 2.88% IN Sodium Hydroxide 25'
Cool the solution in an ice bath and add 1 part of sodium hydroxide to it.
Add 2.5 parts of [and 3.9 parts of 4-furomo-2-methylbutanoic acid chloride (the material prepared in {a} above)]. 3. Mix this mixture at room temperature. After a while, a suspension of 3.75 parts of thiobenzoic acid, 2.4 parts of potassium carbonate and 25 parts of water is added. After concentrating overnight at room temperature, the reaction mixture is acidified with concentrated hydrochloric acid and extracted with ethyl acetate. The organic layer was dried over magnesium sulfate and concentrated under reduced pressure. The residue was chromatographed on a silica gel column using benzene monoacetic acid (7:1) as the eluent, and the fractions containing the desired product were combined and concentrated to dryness under reduced pressure to give 1-(4-benzoylthio- 2-Methylbutanoyl)-L-proline was obtained. Example 7 1-(4-mercapto-2-methylptanoyl)-L-
Production of proline: -1-(4- in the treatment of Example 6)
Using 1-(4-benzoylthio-2-methylbutanoyl)-L-proline obtained in Reference Example 19 in place of 1-(4-benzoylthio-2-methylbutanoyl)-L-proline and treating in the same manner, 1 -(4-mercapto-2-methylbutanoyl)-L-
Got proline. Reference example 20 1-(3-acetylthiobutanoyl)-L-proline.
Preparation of t-butyl ester: -dicyclohexylcarposiimide 6.2 and 3-acetylthiobutyric acid 4.86
2 was added to a solution of L-proline t-butyl ester in 5.1 mL of dichloromethane (60 mL) while stirring in an ice bath. After 1 stir, remove the ice cream and let the mixture stand at room temperature for 1 hour. The precipitate was heated in the oven, the filtrate was concentrated and dried, and the residue was chromatographed on a silica gel column using chloroform as the eluent to obtain the title compound 5.2. Reference Example 21 Production of 1-(3-acetylthiobutanoyl)-L-proline: -1-(3-acetylthiobutanoyl)-L-proline/t-butyl ester (product of Reference Example 20) 5
．． 60% of trifluoroacetic acid and 3% of anisole
0' mixture and keep the solution at room temperature for 1 hour. The solvent was removed under reduced pressure, and the residue was reprecipitated several times from phenol-hexane to give 1-(3-acetylthiobutanoyl)-
L-proline was obtained. This was treated in the same manner as in Reference Example 17 to obtain its dicyclohexylamine salt. Melting point 17
5-17 is 0. Example 8 Preparation of 1-(3-mercaptoptanoyl)-L-proline: -1-(3-acetylthiobutanoyl)-L-proline (product of Reference Example 21)
0.86 methanol 5. Dissolve 20 ml of ammonia solution and keep the reaction mixture at room temperature for 2 hours. The solvent is removed under reduced pressure and the residue is chromatographed on an ion exchange column (Dawex 50) using water as the eluate. Fractions containing the desired product were combined and lyophilized to give 1-(3-mercaptobutanoyl)-L-proline.
．． I got 6 nights. This was treated in the same manner as in Reference Example 17 to obtain its dicyclohexylamine salt. Melting point 183-184
00. Example 9 Preparation of 1,1[-dithiopis(3-propanoyl)]-bis-L-proline: -3-Mercaptopropanoyl-L-proline (0.95%) was dissolved in 20% of water, and the pH was adjusted to 6 with IN sodium hydroxide. Adjust to .5. Add the ethanolic solution of iodine dropwise while carefully adjusting the pH to 6.5 by adding IN sodium hydroxide. When a persistent yellow color appears, the iodine addition is discontinued and the color is eliminated with a small amount of sodium thiosulfate. The reaction mixture is made acidic with concentrated hydrochloric acid and extracted with ethyl acetate. The layer was washed with water, dried, and concentrated to dryness to obtain the title compound. Dicyclohexylamine was added to a solution of this free acid in acetonitrile to obtain its dicyclohexyl ammonium salt. Melting point 179-180q0. Example 101,1'1 [
Dithiopis (21D-methyl-3-propanol)]-
Production of bis-L-proline: Using 3-mercapto-21D-methylpropanoyl-L-broline in place of 3-mercaptopropanoyl-L-proline, the title compound was obtained in the same manner as in Example 9. Ta. Melting point 236-2370. Example 111 Production of 1'-[dithiobis(2-propanoyl)]-bis-L-broline: Produced in the same manner as in Example 9, using 2-mercaptopropanoyl-L-proline instead of -3-mercaptopropanoyl-L-proline. Work-up provided the title compound. Example 12 Production of 1,1'-(dithiobisacetyl)-bis-L-hydroxyproline: -3-mercaptopropanoyl-
The title compound was obtained by treating in the same manner as in Example 9, using 1-(2-mercaptoacetyl)-L-hydroxyproline in place of L-broline. Example 13 1. Preparation of 1'-[dithiobis(4-butanoyl)]-pisu L-proline: 4-mercaptobutanoyl L instead of -3-mercaptoprobanoyl L-proline
The title compound was obtained in the same manner as in Example 9 using -proline. Reference Example 22 Production of 3-[(4-methoxyphenyl)methylthio]-2-methylpropanoic acid - 15.4 liters (0.1 mol) of p-methoxyQ-toluenethiol was added to 8.6 mol of methacrylic acid.
(0.1 mol) is added to a solution of 50% sodium hydroxide. The mixture is heated on a steam stove for 3 hours, then refluxed for 2 hours and cooled. The mixture is extracted with ether, then the aqueous layer is acidified with concentrated hydrochloric acid and extracted with dichloromethane. The acidic extract is washed with brine, dried over magnesium sulphate and evaporated under reduced pressure. The resulting semisolid is dissolved in 50 molar dichloromethane, diluted with 50 molar hexane and cooled. 3-[(4-Methoxyphenyl)methylthio]-2-methylpropanoic acid is collected as a white crystalline solid. Melting point: 74-8〆○ (5.5 degrees). Reference example 23 1-[3-(4-methoxyphenyl)methylthio]-2
-Production of methylpropanoyl-L-proline/rt-butyl ester: -3-[(4-methoxyphenyl)methylthio]-2-methylpropanoic acid 3 obtained in Reference Example 22
．． 6 days (0.015 mol), L-proline, 2.6 hours (0.015 mol) of rt-butyl ester and 3.1 hours (0.015 mol) of dicyclohexylcarbodiimide were dissolved in 50 minutes of dichloromethane, and the mixture was heated at 0°C. Fly for 30 minutes. Remove the cooling rack and let the mixture stand overnight (1 hour).
The resulting suspension is filtered and the filtrate is washed with 5% potassium hydrogen sulfate, saturated sodium hydroxide on carbon and brine, then dried over magnesium sulfate and evaporated under reduced pressure. The resulting clear oil is applied to a 250 silica gel column and chromatographed using 20% ethyl acetate as eluent. Main fraction (Rf=0.70(
On silica gel, evaporate 1-[3-(4-methoxyphenyl)methylthio]-2-methylpropanoyl-L-proline te-butyl ester 5.
．． 5 (93%) was obtained as a clear oil. Rf=0
．． 70 (on silica gel, ethyl acetate); Rf=0.60
(on silica gel, ether). Example 14 1-(3-mercapto-2-methylpropanoyl)-L
- Production of proline: - Estel 1.2 from Reference Example 23
(0.003 mol), anisole 5 and trifluoromethanesulfonic acid 0.5' to trifluoroacetic acid 2
0' under nitrogen and the resulting red solution is left at room temperature for 1 hour. The solution is evaporated under reduced pressure to give a red residue, which is dissolved in ethyl acetate, washed with water, brine, then dried over magnesium sulfate and evaporated. Triturate the residue repeatedly with hexane, evaporate the remaining hexane, and leave no oily residue.
．． Get 4 evenings. A portion of this material (180 females) was subjected to Preparatep thin layer chromatography on a two-ribbed silica gel plate using Bunsen/acetic acid (75:25) as eluent. The main nitroprusside positive zone (Rf=0.40) was collected to obtain 11(3-mercapto-2-methylpropanol)-L-proline 13-9 as an oil. Rf of thin layer chromatography using benzene/acetic acid (75:25) = 0.4 and Rf of thin layer chromatography using chloroform/methanol/acetic acid (50:40:10) = 0. 62. Example 151-(3-mercapto-2-D-methylpropanol)-L
- Production of proline: - Reference example 2 under argon gas seal
1-[3-(acetylthio)-2-D- obtained in 5,27
Methylpropanoyl]-L-proline 10.0 to 1o
Mix in 150 ml of water at o0. Add sodium hydroxide to the mixture and maintain the pH of the solution at 13 for 1.5 hours. After this time the sodium hydroxide has been consumed and the solution is adjusted to pH=2.0 with concentrated sulfuric acid. The aqueous solution is then extracted three times over 150 g of methylene chloride and the combined ethylene chloride fractions are concentrated to an oil. The concentrate is dissolved in ethyl acetate, filtered and the filtrate is diluted with 30' of hexane. The amount of hexane added is 0. Add after 1 hour and then cool the mixture to 1000 °C for 1 hour. The crystals were heated, washed with hexane (2 x 25'), dried to constant weight, and 1-(3-mercapto-21D-methylpropanoyl)-L-proline 6.26% was dissolved in white crystals. obtained as. Melting point 100-1020〇〇Reference example 24 1-[3-tosyloxy-2-methylpropanol]-
Production of L-proline: -Reference Example 1, using the method of wave 1, using 3-acetylthio-2-methylpropanoyl chloride instead of 3-acetylthio-2-methylpropanoyl chloride.
1 using tosyloxy-2-methylpropanoyl chloride
-3[-tosyloxy-2-methylpropanoyl]-L
- Obtained proline. Reference Example 25 Production of 1-[3-acetylthio-2-D-methylpropanol]-L-proline: 1-[3 obtained in Reference Example 24]
-Tosyloxy-2-methylpropanoyl] 3.5 parts of L-proline are added to a solution of 1.14 parts of thiol acetic acid and 3.5 parts of trietheramine in 20 parts of ethyl acetate. The solution is kept at 50°C for 3 hours, cooled, diluted with 100' of ethyl acetate, and washed with dilute hydrochloric acid. The organic layer is dried and concentrated to dryness under reduced pressure. Dissolve the remaining port in acetonitrile and add dicyclohexylamine. The crystal precipitate was recrystallized from isopropanol to give 1-[3-acetylthio-2
-○-Methylpropanoyl]-L-proline dicyclohexylamine salt was obtained. Melting point 187-18 is ○. [Q
] Volume -670 (in ethanol, concentration c=1.4). This salt is converted to the free acid. Melting point: 83-860 (melting point: 10 when seeds of a substance with a high melting point are added to a solution containing a crystalline substance)
4 to 105 qo isomorphs are obtained). Example 16 Preparation of 1-(3-mercaptopropanoyl)-L-proline: -1-[3-[(ethylamino)carbonyl]
Thio]propanoyl]-L-proline 75 female (0.27
mmol) is dissolved in 1 part of concentrated ammonium hydroxide and 1 part of water, and the solution is left at room temperature for 1 hour under an argon atmosphere. Dilute the solution with a little water and extract with ether. The aqueous layer is made acidic with cold concentrated hydrochloric acid and extracted with ethyl acetate. The combined extracts are dried and concentrated under reduced pressure to yield a compound corresponding to the product of Example 3. Thin layer chromatography (on silica gel, benzene:acetic acid (7:3) Rf=0.4 Reference example 2
6 Preparation of methacryloyl-L-proline: Dissolve 23.0 molar (0.2 mol) of -L-proline in 100 m of water,
Worshiping flies in an ice bath. Methyl isobutyl ketone and 25 parts of methacryloyl chloride (0.2 mol) are added dropwise over a period of 3 hours.
The pH of the reaction mixture is maintained at 0.7 and sodium hydroxide solution is added at the same time. The base addition is continued for 4 hours after the acid chloride addition is complete. The reaction mixture was adjusted to pH 5 with concentrated hydrochloric acid and extracted with ethyl acetate. The aqueous layer is then adjusted to pH 2.5 and thoroughly extracted with ethyl acetate. The acidic extract is washed with brine and dried over magnesium sulfate. Treat the ethyl acetate solution with 40% dicyclohexylamine and cool overnight. The obtained white precipitate was filtered and dried to obtain a white solid 29 with a melting point of 202 to 21,000.
Evening (39%). This solid is crystallized from 1500 ml of acetonitrile/isopropanol (3:1) to give 19.7 ml of methacryloyl-L-proline dicyclohexylamine salt as fine white needles. Melting point 20
2 to 210 qo. Dissolve this salt in water/ethyl acetate and acidify the mixture with concentrated hydrochloric acid. The resulting suspension is filtered to remove a fine white precipitate and washed thoroughly with ethyl acetate.
Saturate the filtrate with sodium chloride and thoroughly extract with ethyl acetate. The extract is washed with brine, dried over magnesium sulfate, and evaporated to a clear oil that solidifies. Crystallization from ethyl acetate/hexane gave 7.5 g (85%) of methacryloyl L-proline as a white crystalline solid. Melting point 89-93oo. A pure product was obtained by recrystallization. Melting point: 95-980〇〇Reference example 271-(3-acetylthio2-D-methylpanoyl)-L-proline production: - Methacryloyl-L-proline 183 female (0.001 mol) obtained in Reference example 26 was dissolved in thiol acetic acid 0 Dissolve in 50% diluted solution and leave at room temperature for 1 hour. The solution is evaporated under reduced pressure to give a yellow residue. 240 m9 of a clear oil are isolated as the main fraction by prevarateb thin layer chromatography (on silica gel, dichloromethane/metal/acetic acid (90:5:5)). Thin layer chromatography (dichloromethane/methanol/acetic acid (
90:5:5)), and this substance was found to be 1-(3-acetylthio-2-DL-methylpropanoyl)-1-L-proline, which corresponds to the product of Reference Example 1. Rf!
0.35. (Benzene/acetic acid (75:25)) Rf=0
．． 38. Dissolve this oil in 3 parts of acetonitrile,
Treat with dicyclohexylamine until the solution is basic and cool. Collect 106 female white crystalline solids. Melting point 175-18ro
. Crystallized from isopropanol, melting point 187-1 total 0○
1-(3-acetylthio-2-D-methylpropanoyl)-L-proline dicyclohexylamine salt was obtained. This product corresponds to that of Reference Example 12A. Example 17 Production of 1,1'-[dithiobis-(2-methyl-3-propanol)]-bis-L-proline: - Using the method of Reference Example 1, 3,3'-dithiobis was used instead of 3-acetylthio-2-methylpropanoic acid. Using -2-methylpropanoic acid, 1,1'-[dithiobis-(2-methyl-3
-propanoyl)]-bis-L-proline is obtained. Example 18 1-(3-mercapto-2-methylbropanoyl)-L
- Preparation of proline: - Add 10.09 g of zinc dust to a slurry of 5.0 g of the product of Example 17 and 100 g of 1.0 N sulfuric acid and stir this mixture for 4 hours at 18 o'clock under nitrogen gas blanket. The solution is filtered, the zinc is washed with 20 portions of water, the furnace liquors are combined and extracted with methylene chloride (3 x 75 ml). Back-extract with water (25 cm) with a ethylene chloride wash and then concentrate the organic solution to an oil. This oil was dissolved in 20% ethyl acetate and filtered. Add 15' of hexane to the furnace liquid,
Stir this mixture for 15 minutes. Then, an additional 30' of hexane is added and the solution is cooled to 5° C. for 1 hour.
The mixture was filtered and the product was extracted with hexane (2 x 10'
) and drying to give the product 1-(3-mercapto-2
-Methylpropanoyl)-L-proline white crystals4.
I got it on the 17th. Thin layer chromatography, Rf=0.6
0 (solvent system: benzene/acetic acid (75:25)). Reference Example 28 Production of 3-benzylthio-2-methylpropanoic acid: - 3-benzylthio-2-methylpropanoic acid is obtained by the method of Reference Example 22 using Q-toluenethiol instead of p-methoxyQ-toluenethiol. Reference Example 29 Production of 1-[3-(benzylthio)-2-methylpropanoyl]-L-broline tert-butyl ester:-
By the method of Reference Example 23, 3-[(4-methoxyphenyl)
Reference Example 2 instead of methylthio]-2-methylpropanoic acid
Using the 3-pendylthio-2-methylpropanoic acid obtained in 8, rt-butyl ester of 1-[3-(benzylthio)-12-methylpropanoyl]-L-proline is obtained. Reference Example 30 Production of 1-[3-(benzylthio)-2-methylpropanoyl]-L-proline: -1-[3-(benzylthio)-2-methylpropanoyl]-L-broline obtained in Reference Example 29. Add 7.8 parts of rt butyl ester into a mixture of 55 parts of anisole and 110 parts of trifluoroacetic acid. After standing at room temperature for 1 hour, the solvent was removed under reduced pressure and the residue was dissolved in ether, washed several times with saturated sodium chloride, dried over magnesium sulfate and evaporated to dryness under reduced pressure to give 1-[3
-(benzylthio)-2-methylpropanol]-L-
Got proline. Rfo. 5 (on silica gel, benzene/acetic acid (3:1)
) Rfo. 5 (on silica gel, methyl-ethyl ketone/
Acetic acid/pyridiso/water (14:1:2:1)). Example
191-(3-mercabuto-2-methylpropanoyl)
-Production of L-proline: -1-[3- obtained in Reference Example 30
0.1 of (benzylthio)-2-methylpropanoyl]-L-proline is suspended in 10 parts of boiling liquid ammonia and small pieces of sodium are added with stirring until a permanent blue color is formed. The color is decolorized with a few crystals of ammonium sulfate and the ammonia is evaporated under a stream of nitrogen. Dissolve the remaining bamboo in a mixture of dilute hydrochloric acid and ethyl acetate. The organic layer was dried and concentrated under reduced pressure to obtain 1-(3-mercapto-2-methylpropanoyl)-L-proline. Rfo. 35 (on silica gel, benzene/acetic acid (3:1)), Rfo. 5 (on silica gel, mether/ethylketone/acetic acid/pyridine/water (14:1:2:1)), corresponding to the compound of Example 5. Reference Example 31 Production of 1-[3-(triphenylmethylthio)-2-methylpropanol]-L-proline: -3 triphenylmethylthio 2-methylpropanoic acid 1.8% and N,N
-Carponyldiimidazole is dissolved in 10 parts of tetrahydrofuran at room temperature with 0.8 parts of water. After 2 minutes, this solution is added to a mixture of 0.6 parts L-proline, 1 part N-methylformoline, and 20 parts dimethylacetamide. The resulting mixture was allowed to stand overnight at room temperature, concentrated to dryness, and the residue was dissolved in a mixture of ethyl acetate and 10% aqueous potassium hydrogen sulfate solution. The organic layer was separated, dried, and concentrated to dryness under reduced pressure to obtain 1-[3-(triphenylmethylthio)-2-methylpropanoyl]-1L-proline. Rfo. 4(
on silica gel, benzene/acetic acid (3:1)), Rfl.
0 (on silica gel, methyl/ethyl ketone/acetic acid/pyridine/water (14:1:2:1)). Reference Example 32 Preparation of 3-(tetrahydropyran-2-ylthio)-12-methylpropanoic acid: 2.4 hours of -3-mercapto-2-methylpropanoic acid and 1.9 hours of freshly distilled 2,3-dihydro-saki-pyran. Add 2.8 g of boron trifluoride ether rust compound to 60 g of benzene. After 2 hours, 4 tablespoons of potassium carbonate are added and the mixture is filtered. The furnace liquid was concentrated to dryness to obtain 3-(tetrahydropyran-2-ylthio)-12-methylpropanoic acid. Reference example 33 1-[3-(tetrahydropyran-2-ylthio)-2
-Methylbropanoyl]-L-proline production: -By the method of Reference Example 31, 31(tetrahydropyran-2-ylteo)-12 obtained in Reference Example 32 was used instead of 3-triphenylmethylthio-2-methylpropanoic acid. -Methylpropanoic acid was used to obtain 1-[3-(tetrahydropyran-2-ylthio)-2-methylpropanol]-L-proline. Rfo. 8 (on silica gel, methyl-ethyl ketone/acetic acid/pyridine/water (14:1:2:1)). Example 2
0 1-(3-mercapto-2-methylpropanoyl)-L
-Production of proline: -Dissolve 1 part of 1-[3-(tetrahydropyran-2-ylthio)-2-methylpropanoyl]-L-broline obtained in Reference Example 33 in a mixture of 2 traces of methanol and 25 parts of concentrated hydrochloric acid. and this solution was heated at room temperature for 30
Leave for a minute. The solvent was removed under reduced pressure to yield 1-(3-mercapto-2-methylpropanoyl)-L-proline. Rfo. 35
(on silica gel, benzene/acetic acid (3:1)), Rfo
．． 5 (on silica gel, methyl-ethyl ketone/acetic acid/pyridine/water (14:1:2:1)) corresponds to the compound of Example 5. Reference Example 34 Production of acetamidomethylthio-2-methylpropanoic acid: 2.4 hours of -3-mercapto-2-methylpropanoic acid and 1.8 hours of N-hydroxymethylacetamide are dissolved in trifluoroacetic acid, and this solution is stored at room temperature for 1 hour. . The trifluoroacetic acid was removed under reduced pressure and the residue was dried over potassium hydroxide under reduced pressure to give 3-acetamidomethylthio-2.
-Methylpropanoic acid was obtained. Reference Example 35 Production of 1-[3-(acetamidomethylthio)-2-methylpropanoyl]-1L-proline: - By the method of Reference Example 33, 3-(tetrahydropyran-2-ylthio)-2
- Using 3-acetamidomethylthio-2-methylpropanoic acid obtained in Reference Example 34 instead of methylpropanoic acid, 1
-[3-(acetamidomethylthio)-2-methylpropanoyl]-L-proline was obtained. Rfo. 2 (on silica gel, benzene/acetic acid (3:1)
), Rfo. 3 (on silica gel, methyl-ethyl ketone/acetic acid/pyridine/water (14:1:2:1)). Example 21 1-(3-mercapto-2-methylpropanoyl)-L
-Production of proline: 1-[3-(acetamidomethylthio)-2-methylpropanoyl]- obtained in Reference Example 35
1.4 units of L-proline and 1.93 units of mercury acetate were added to 25 units of acetic acid.
Dissolve in a mixture of 25 parts of earth and water. After one hour on the steamer, hydrogen sulfide is bubbled in until no more mercury sulfide precipitates are visible. The mixture was heated in a furnace, the precipitate was washed with ethanol, and the furnace solution was concentrated to dryness under reduced pressure to obtain 1-(3-mercapto-2-methylpropanoyl)-L-proline. Rfo. 35 (on silica gel, benzene/acetic acid (3:1)), Rfo. 5 (on silica gel, methyl-ethyl ketone/acetic acid/pyridine/water (1
4:1:2:1)) corresponds to the compound of Example 5. Reference example 36 1-(3-mercapto-2-methylpropanoyl)-L
-Production of proline ten butyl ester: 1.2 units (10 mmol) of -3-mercabuto-2-methylpropanoic acid and 1.7 units (10 mmol) of L-proline ten nine butyl ester.
To a cold (5° C.) solution of 2.5 μl of dicyclohexylcarbodiimide (2.26 μl) of dichloromethane is added in portions. After 2 hours at room temperature, 5 drops of acetic acid are added and the mixture is filtered and evaporated to an oily residue. This residue is dissolved in 20 kg of petroleum ether-ethyl acetate (3:1) and applied to a 150 ml silica gel column prepared with petroleum ether. The fractions separated with petroleum ether-ethyl acetate (1:1) contain the 1-(3-mercapto-2-methylpropanoyl)-L-proline rt-butyl ester product. 0.6 minutes of this fraction is dried over pentoxide under reduced pressure for 1 hour. Rfo. 6 (on silica gel, benzene/acetic acid (3:1)
)], Rfo. 8 (on silica gel, methyl-ethyl ketone/acetic acid/pyridine/water (14:1:2:.)). Example 22 1-(3-mercapto-2-methylpropanoyl)-L
- Production of proline: - Using the method of Example 3C, 1-(3-mercapto-2-methylpropanoyl) obtained in Reference Example 36 was used instead of 1-(3-mercaptopropanoyl)-L-proline. 1-(3-mercapto-2-
methylpropanol)-L-proline was obtained. Rfo. 35 (on silica gel, benzene/acetic acid (3:1
)), Rfo. 5 (on silica gel, methyl-ethyl ketone/acetic acid/pyridine/water (14:1:2:1)) corresponds to the compound of Example 5. The racemic form of the final product of any of the preceding examples is made using the DL form of the starting amino acid instead of the L form. Similarly, the D-form of the final product in any of the previous examples is made using the D-form of the starting amino acid rather than the L-form. Reference Example 37 100 mirrors containing 9 parts of 100 each of 1-(2-lcaptopropanoyl)-L-proline are prepared from the following ingredients. 1-(2-captopropanoyl)-L-proline
100 cornstarch 50 tazelatin
7.5 evening Azeser (Avic)
el) (microcrystalline cellulose) 25% magnesium stearate 2.5% 1-(2-mercaptopropanoyl)-L-proline and cornstarch are mixed with an aqueous solution of gelatin. This mixture is dried and ground to a fine powder. Avicel and then magnesium stearate are mixed and granulated. This was then compressed into tablets to obtain 100 tablets each containing 100 mg of active ingredient. Reference Example 38 In Reference Example 37, 1-(3-mercapto-2-D-methylpropanoyl)-L-proline was used instead of 1-(2-〆captopropanoyl)-L-proline, and 1 -(3-mercapto-2-○-methylpropanoyl)-L-proline, 100 samples each containing 9 of 100 were prepared. Reference Example 391-(2-Mercaptoacetyl)-L-Proline 100 pieces each containing 9 parts of 200 were prepared from the following ingredients. 1-(2-mercaptoacetyl)-L-proline
200 yen Lactose 100 yen Avicel
150 yen corn starch
50 Magnesium tastearate 5-11 (2-mercaptoacetyl)
- Mix L-proline, lactose and Avicel;
Then mix in the cornstarch. The dry mixture was compressed in a tablet machine to obtain 100 tablets of 505 tablets each containing 200 parts of the active ingredient. MethoCel containing lake containing yellow #6 tablets as pigment
The tablets were coated and formulated with a solution of E15 (methylcellulose). Reference Example 40 Two-piece #1 gelatyl capsules each containing 250 parts of 1-(2-mercaptopropanoyl)-L-proline are filled with a mixture of the following ingredients. 1-(2-captopropanoyl)-L-proline
250 9 Magnesium Stearate 7 9 USP Lactose 193 9 Reference Example 41 An injection solution was prepared as follows. 1-(2-〆kabutopropanoyl)-L-proline
500 female Methylparaben 5 Propalparaben 1 Ta Sodium chloride
Add 25 ta of water for injection
Dissolve 5 volumes of active substance, preservative and sodium chloride in 3 volumes of water for injection, then make up to 5 volumes. This solution is heated in a sterile furnace, aseptically filled into pre-sterilized vials, and then sealed with pre-sterilized rubber stoppers. Each vial contains 5' of solution at a concentration of 100 M of active ingredient per injection. Reference example 42 Reference example using 1,r-[dithiopis(21D-methyl-3-propanoyl)]-pis-L-proline 100%, a substitute for 1-(2-〆captopropanoyl)-L-proline 37 to obtain 1,1′-[dithiobis(2
100 pseudo-sodiums containing 9 parts of 100 each of (1D-methyl-3-propanoyl)]-bis-L-broline were prepared. Reference Examples 37, 33, 40 or 41 were replaced with the products obtained in the Examples of the respective production methods described above, and formulations were prepared in the same manner.

Claims (1)

[Scope of Claims] 1 Formula: ▲There are mathematical formulas, chemical formulas, tables, etc.▼ [In the formula, R_1 and R_4 each represent hydrogen or lower alkyl. R_2 represents hydrogen or a group represented by the formula: ▲There are numerical formulas, chemical formulas, tables, etc.▼. n represents 0, 1 or 2. ] and its salts. 2. The compound according to claim 1, wherein R_4 is hydrogen. 3. The compound according to claim 2, wherein n is 1. 4. The compound according to claim 2, wherein R_2 is hydrogen. 5. The compound according to claim 2, wherein R_1 is hydrogen or methyl. 6. The compound according to claim 1, wherein proline in formula [I] is an L-isomer. 7. The compound according to claim 1, which is 1-(3-mercapto-2-D-methylpropanoyl)-L-proline. 8. The compound according to claim 1, which is 1,1'-[dithiobis(2-D-methyl-3-propanoyl)]-bis-L-proline.