JPH0430950B2 - - Google Patents

Description

[Detailed description of the invention]

Industrial Application Field The present invention relates to enkephalinase inhibitors, more specifically, novel mercaptoalkanoyl and acylmercaptoalkanoyl compounds having enkephalinase inhibitory activity and useful as analgesics, and pharmaceutically acceptable compounds thereof. Concerning salts. Structure and Effects of the Invention The novel compound of the present invention is represented by the following formula []. [In the formula, R ₁ is hydrogen or

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【formula】

[Formula] R ₃ is hydrogen, lower alkyl,

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【formula】

[Formula] or -(CH ₂ ) _p -cycloalkyl, R ₄ is hydroxy, lower alkoxy,

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【formula】

[Formula] −O−(CH ₂ ) [ _p+1 ] − OH,

[expression] or

[Formula] n is an integer from 1 to 15, R ₅ is lower alkyl,

【formula】

【formula】

【formula】

[Formula] Ma Taha

[Formula] p is an integer of 1 to 4, m is 0 or an integer of 1 to 4, R ₆ is lower alkyl having 1 to 4 carbon atoms, 1 carbon number
~4 lower alkoxy, lower alkylthio having 1 to 4 carbon atoms, hydroxy, Cl, Br, F, amino, -
NH-lower alkyl (1 to 4 carbon atoms), -N (lower alkyl) ₂ (lower alkyl has 1 to 4 carbon atoms), nitro or trifluoromethyl, r is an integer from 1 to 3 (provided that R ₆ is Hydroxy,
r is 1 only for methyl, methoxy, Cl or F
), R ₁₀ is hydrogen, lower alkyl, cycloalkyl or phenyl, R ₁₁ is hydrogen, lower alkyl, lower alkoxy or phenyl, and R ₇ and R ₈ are each independently hydrogen, lower alkyl,

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【formula】

【Formula】also For details

[Formula] or R ₇ and R ₈ are both combined with the N-atom to form the formula: (q is 0 or 1, A is CH-R ₉ , oxygen or N-R ₉ , and R ₉ is hydrogen or lower alkyl having 1 to 4 carbon atoms)] Definitions of various symbols The phrase used is “lower alkyl”
refers to a straight chain or branched hydrocarbon group having 7 or less carbon atoms, such as methyl, ethyl, propyl, isopropyl, isobutyl, butyl, t-butyl, pentyl, isopentyl, and the like.
Preferred lower alkyls have 4 or less carbon atoms,
Most preferred are methyl and ethyl. Similarly the phrases "lower alkoxy" and "lower alkylthio"
refers to such an alkyl bonded to oxygen or sulfur. The term "cycloalkyl" refers to a saturated ring having 3 to 7 carbon atoms, with cyclopentyl, cyclohexyl and cycloheptyl being preferred. formula:

【formula】

[expression] and

[Formula] indicates that the alkylene bridge is bonded to an available carbon atom. US Pat. No. 4,401,677 to Greenberg et al.
Various mercaptoalkanoyl alpha-amino acids have been disclosed to be useful as analgesics based on their enkephalinase inhibitory activity. US Pat. No. 4,053,651 to Ondetsutei et al.
It has been disclosed that various mercaptoalkanoyl and acylmercaptoalkanoyl α-amino acids are useful as antihypertensive agents based on their angiotensin-converting enzyme inhibitory activity. Loukies et al., “Nature” (vol. 288, November 1980)
May, pp. 286-288), thiorphan, [(D,L)-3-mercapto-2-benzylpropanoyl]-glycine, can be administered at nanomolar concentrations in vitro and intracerebroventricularly or systemically. It has been disclosed that it is an inhibitor of enkephalinase in vivo. European Patent Application No. 38758 to Roukies et al. discloses that various α-amino acid derivatives, including mercaptoalkanoyl and acylmercaptoalkanoyl derivatives, have enkephalinase inhibitory activity. “Biochemical and
Biophysical Research Comm.” (Volume 109, No. 4,
(1982, pp. 1303-1309) discloses that various substituted N-carboxymethyl dipeptides (including those having a terminal β-alanine group) have enkephalinase inhibitory activity. European Patent Application No. 54862 to Berger et al. describes the formula: (R ₂ is a carboxylic or phosphonic acid or ester residue, and y is 0 or an integer from 1 to 3) are disclosed. “Biochemistry” by Kutsyuman et al. (Vol. 16, No.
25, 1977, pp. 5484-5491) disclose various carboxyalkanoyl and mercaptoalkanoyl amino acids as angiotensin converting enzyme inhibitors, these compounds having the formula: It is indicated by. Sandeen et al. US Pat. No. 4,235,885 and
No. 4297275 has the formula: (m is 0 or an integer from 1 to 9, R ₃ includes hydrogen,
and acylmercaptoalkanoyl compounds, where R ₄ includes hydroxy and amino, are disclosed. US Pat. No. 4,327,111 to Sundeen et al.
formula: (R ₄ is hydrogen, alkyl or aryl, n is 1
Mammalian collagenase inhibitors are disclosed comprising mercaptoalkanoyl and acylmercaptoalkanoyl compounds, wherein R is an integer of 20 to 20, and R ₃ is alkyl of 3 to 8 carbon atoms, cycloalkyl of 3 to 7 carbon atoms, aryl or arylalkyl). There is. The compound of the present invention of formula [] has the formula: The formula for the amino compound is: or its chemical equivalent. Due to the above acylation, R ₁ becomes

An acylmercaptoalkanoyl compound of the formula [] is obtained. Treatment of these acylmercaptoalkanoyl compounds by conventional hydrolysis provides the corresponding mercaptoalkanoyl compounds of formula [ ] in which R ₁ is hydrogen. This acylation reaction can be carried out in the presence of a coupling agent (such as dicyclohexylcarbodiimide) or by the formation of an acid of formula [] to its mixed anhydride, symmetrical anhydride, acid halide, active ester or using Woodward's reagent K. , i.e. n-ethoxycarbonyl-2-ethoxy-1,2
- Can be activated using dihydroquinoline or the like. For a discussion of acylation methods, see Methoden der Organischen by Houben-Veil.
Chemie” (Vol.XV, Part, 1 page or less, 1974
See ). Preferably, acid halides (especially acid chlorides) of the formula [] are used. The product of formula [] where R ₁ is hydrogen and R ₄ is lower alkoxy or various substituted lower alkoxy,
R ₁ is

A compound of the formula [Formula] in which R ₄ is hydroxy can be prepared by treating with a suitable alcohol in the presence of dicyclohexylcarbodiimide and dimethylaminopyridine followed by selective saponification. R ₁ is

[Formula] The product of formula [] where R ₄ is hydroxy can be produced by acylating the corresponding compound of formula [] where R ₁ is hydrogen and R ₄ is hydroxy with an appropriate acyl halide. . R ₄ is

A compound of formula [] which is [formula] can be produced by using an amino compound of formula [] having an amide group at a predetermined position. In addition, when R ₇ and R ₈ are hydrogen, the acylation reaction can be completed using an intermediate of formula [] where R ₄ is lower alkoxy as a carboxylic acid ester, and the acyl mercaptoalkanoyl amino acid is converted into a methanol solution of ammonia. Process with. _R7 and _R8
are not necessarily both hydrogen, then R ₁ is

[Formula] A compound of formula [] in which R ₄ is hydroxy is prepared in the presence of dicyclohexylcarbodiimide as follows: can be reacted with other amines. Then with the above hydrolysis

[Formula] can be eliminated to obtain a compound of formula [] in which R ₁ is hydrogen. R ₄ is

The ester product of formula [], which is [formula], can be produced by using an amino compound of formula [] having an ester group at a predetermined position in the above reaction. Also R ₄
is hydroxy, R ₁ is

The product of formula [] which is [formula] is the formula: (L is a leaving group such as chlorine, bromine, tolylsulfonyl, etc.). next

The [Formula] group can be removed by selective saponification to yield products in which R ₁ is hydrogen. Amino intermediates of formula [] are known in the art,
This is based on known α-amino acids in Andetsutei et al.'s "Journal of Medicinal Chemistry" (vol. 18,
No. 7, 1975, pp. 761-763). Acylmercaptocarboxylic acid intermediates of formula [] are prepared according to procedures described in various patent documents and references (e.g., Ondetsutei et al.
See Patents Nos. 4,053,651 and 4,105,776, and Sundeen US Pat. Nos. 4,235,885 and 4,327,111). Preferred compounds of formula [] are R ₁ is hydrogen,

[expression] or

[Formula] (especially hydrogen), R ₂ is

【formula】

【formula】

【formula】

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【formula】

[expression] or

[Formula] (m is 1 or 2, and R ₆ is methyl, methoxy, Cl, F, amino), nitro or hydroxy), especially R ₂ is benzyl, R ₃ is hydrogen, straight chain with 1 to 4 carbon atoms or branched lower alkyl,

【formula】

【formula】

【formula】

【formula】

【formula】

[expression] or

[Formula] (m is 1 or 2, and R ₆ is methyl, methoxy, Cl, F, hydroxy, amino or nitro), R ₄ is hydroxy, methoxy, -NH ₂ or

[Formula] (R ₁₀ is hydrogen, straight chain or branched lower alkyl having 1 to 4 carbon atoms or cyclohexyl, and R ₁₁ is straight chain or branched lower alkyl having 1 to 4 carbon atoms), especially when R ₄ is hydroxy, and n is an integer from 1 to 9, especially from 3 to 6. The compound of formula [] has one or two asymmetric centers (two asymmetric centers when R ₃ is other than hydrogen). Therefore, the compound of formula [] exists in three-dimensional forms or in racemic mixtures thereof. All of these shapes can be used in the method of the invention. In the above-mentioned syntheses, the starting compounds can be used in the form of racemic mixtures or as single isomers. Compounds of formula [] where R ₄ is hydroxy are:
Forms basic salts with various inorganic or organic bases. Pharmaceutically acceptable salts of compounds of formula [] are useful in the methods of the invention. Such pharmaceutically acceptable salts include alkali metal salts such as sodium or potassium, alkaline earth metal salts such as calcium or magnesium, and salts derived from amino acids such as arginine and lysine. Such salts are
It is produced by reacting the acid form of the compound (ie, R ₄ is hydroxy) with a base that provides an equivalent amount of a given basic ion in the medium in which the salt is precipitated or in an aqueous medium, followed by freeze-drying. In addition, compounds of formula [ ] having an amine group form acid addition salts with various inorganic and organic acids. Such pharmaceutically acceptable salts include:
For example, hydrohalides (hydrochlorides, bromates, etc.), sulfates, phosphates, nitrates, arylsulfonate salts (camphor sulfonate salts, benzenesulfonate salts, toluenesulfonate salts, etc.), citrates, ascorbates. , maleate, fumarate, pamoate, acetate, tartrate, salicylate, etc.). To isolate such compounds, it is often convenient to form the acid salt and precipitate it in a medium in which it does not dissolve. These bases also form quaternary ammonium salts with quaternizing agents, and the quaternizing agents are those that are acceptable for pharmaceutical use, such as lower alkyl halides (methyl chloride, methyl bromide, methyl chloride). ), lower alkyl sulfates (methyl sulfate, ethyl sulfate, etc.), monocyclic aryl (lower alkyl) halides and sulfates (benzyl chloride, benzyl sulfate, etc.). Formation of this quaternary ammonium salt is accomplished by reacting the base with an alkyl halide, sulfate, and the like. The compound of formula [] is useful as an analgesic based on its enkephalinase inhibitory activity when administered to mammals. Although the scope of the present invention is not limited to any particular theory or mechanism of action, endogenous sedative pentapeptides,
That is, [Met ⁵ ]-enkephalin (Tyr-Gly-Gly
-Phe-Met) and (Leu ⁵ )-enkephalin (Tyr-Gly-Gly-Phe-Leu) are neurotransmitters necessary for central pain mediation [Hyuges et al., Nature, Vol. 258, 1975] December, pp. 577-579)
], and these sedative peptides are functionalized by cleavage of the Gly ³ -Phe ⁴ peptide bond by a specific peptidar (enkephalinase, which is presumed to be located specifically at nerve terminals in the brain from which enkephalins are released). to be inactivated [Malfroy et al., “Nature”, vol. 276, November 1978, 523-
526)] has been proposed. This specific inhibitor of enkephalinase enhances the regeneration of endogenous enkephalin released from isolated brain slices [Patey et al., Science, vol. 212, June 1981, 1153
The sedating antagonist naloxone renders reversible mice analgesic [see Loukies et al., supra]. Prolonging the body's natural analgesic action released from peripheral and central sites provides pain relief as well as other medicinal effects such as antitussive or antidiarrheal activity. Thus, by administering a composition containing the compound of the present invention or one or a mixture of its pharmaceutically acceptable salts, the suffering of the mammalian host is alleviated. Approximately 0.1-100mg/body weight (Kg)/
daily, preferably at a basic dose of about 1 to 25 mg/body weight (Kg)/day, or preferably 2
~4 divided daily doses produce the desired analgesic activity. Although the analgesic compositions of the invention are preferably administered orally, parenteral routes such as subcutaneous injection may also be used. In particular, the compound of the present invention [] has a strong enkephalinase inhibitory activity, but it is far more effective as an angiotensin converting enzyme inhibitor than the corresponding α-amino acid compound described in the patent documents of Ondetsutei et al. and Greenberg et al. It was found that the activity was low. Therefore, by administering the compound of the present invention [], a selective analgesic effect that is not possible with conventional α-amino acid compounds can be obtained. The compound of the present invention [] can be administered in a dosage form such as a tablet, capsule, or elixir for oral administration.
For parenteral administration, it can be used as an enkephalinase inhibitor for alleviating pain by formulating it as a sterile solution or suspension. About 10 to 500 mg of a compound of the invention [ ] or a physiologically acceptable salt or mixture thereof, in a unit dosage form as required by accepted pharmaceutical practice, in a physiologically acceptable vehicle, carrier, excipient, Prepared with binders, preservatives, stabilizers, fragrances, etc. The amount of active ingredient in these preparations or combinations is determined so as to obtain a suitable dosage within the above-mentioned range. Specific examples of adjuvants that can be mixed into tablets, capsules, etc. include the following. Binders such as gum tragacanth, gum acacia, corn starch or gelatin, excipients such as dicalcium phosphate, disintegrants such as corn starch, potato starch, alginic acid, lubricants such as magnesium stearate, sucrose, lactose or saccharin, etc. sweeteners, aromatics such as peppermint, wintergreen oil or cherry oil. When the dosage unit form is a capsule, it may contain, in addition to the materials enumerated above, a liquid carrier such as a fatty oil. Various other substances may also be present as coatings or otherwise to modify the physical form of the unit dosage form. For example, tablets may be coated with shellac, sugar or both. A syrup or elixir may contain the active compound, sucrose as a sweetening agent, methyl and propylparabens as preservatives, a dye and flavoring such as thierry or orange flavor. Sterile injectable compositions are prepared in accordance with normal pharmaceutical practice by incorporating the active substance into a suitable vehicle, such as water for injection, naturally occurring vegetable oils (sesame oil, coconut oil, peanut oil, cottonseed oil, etc.), or synthetic fatty vehicles (ethyl oleate, etc.). It can be blended by dissolving or suspending it in (e.g.). Further, a buffer, a preservative, an antioxidant, etc. can be mixed as necessary. The following examples are specific examples of the invention. In the examples, the temperature unit is °C. Example 1 Production of (±)-3-[[2-(mercaptomethyl)-1-oxo-3-phenylpropyl]amino]propanoic acid: - a (±)-3-[[2-[(acetylthio ) methyl]
-1-oxo-3-phenylpropyl]amino]
Production of propanoic acid ethyl ester: - (±)-2-[(acetylthio)methyl]-3-phenylpropanoic acid 0.95 g (4.0 mmol) in 8 ml of ethyl ether solution in a drying tube, oxalyl chloride
0.37 ml (4.2 mmol) and then carefully treated with a catalytic amount of dimethylformamide (3 drops).
The mixture is stirred at room temperature for 1 hour and concentrated under reduced pressure. Add 5 ml of tetrahydrofuran to the residue,
Drive off any remaining oxalyl chloride under reduced pressure. The two-layer (yellow and green) liquid residue is treated with 5 ml of methylene chloride and the soluble portion is decanted into the dropping funnel. This solution was mixed with β-alanine ethyl ester hydrochloride in 10 ml of methylene chloride under a nitrogen atmosphere.
Add dropwise over 10 minutes to a cold (ice/methanol) mixture of 0.61 g (4.0 mmol) and 1.46 ml (8.38 mmol) diisopropylethylamine. After stirring for 3 hours, the resulting slurry is filtered and concentrated. The residue was treated with 50 ml of ethyl acetate, filtered again and the solution was dissolved in 10% potassium bisulfate, water, saturated sodium bicarbonate and 50% brine (30 ml each).
, dried over sodium sulfate and concentrated under reduced pressure. 1.20 g of a viscous yellow oily residue is purified on a column of 80 g of silica gel (230-400 mesh) eluting with hexane/acetone (7:2). Fractions containing the desired product (TLC) are combined and concentrated;
As a clear, almost colorless oil (±)-3-
[[2-[(acetylthio)methyl]-1-oxo-
1.04 g of 3-phenylpropyl]amino]propanoic acid ethyl ester is obtained. b (±)-3-[[2-(mercaptomethyl)-1
- Preparation of oxo-3-phenylpropyl]amino]propanoic acid: - A solution of 1.02 g (3.02 mmol) of the product obtained in section a above in 6 ml of methanol is cooled (ice/methanol) and added to it for 1N under nitrogen atmosphere. Sodium hydroxide
Add 6.35ml over 15 minutes. The mixture is stirred for 3 hours while warming to room temperature (TLC shows incomplete reaction) and an additional 3.0 ml of 1N sodium hydroxide is added in one portion. After stirring for a further 3 hours at room temperature, the mixture is concentrated under reduced pressure to approximately 1/2 volume and the residue is partitioned between 50 ml of water and 30 ml of ethyl acetate. Wash the aqueous layer with 30 ml of ethyl acetate, adjust the pH to about 2 with concentrated hydrochloric acid, and extract again with ethyl acetate (20 ml x 3). The combined organic layers were washed with water and brine (30 ml each), dried over sodium sulfate and concentrated under reduced pressure to give (±)-3-[[2-(mercaptomethyl)-1] as a white solid. -oxo-3-phenylpropyl]
0.71 g of amino]propanoic acid was obtained. Melting point 76-82
℃. TLC (on silica gel, toluene/acetic acid (4:
1) R _f =0.37; small spots at 0.07. Elemental analysis, calculated as C ₁₃ H ₁₇ NO ₃ S: C, 58.40%; H, 6.41%; N, 5.24
%; S, 11.99%; Actual value: C, 58.17%; H, 6.34%; N, 5.15
%; S, 11.85%. Example 2 (±)-4-[[2-(mercaptomethyl)-1-
Preparation of oxo-3-phenylpropyl]amino]butanoic acid: - a Preparation of 4-aminobutanoic acid methyl ester hydrochloride: - Under a nitrogen atmosphere, 4.12 g (40 mmol) of 4-aminobutanoic acid in 50 ml of methanol was dissolved in cold (- 10℃) Add 5.86 ml (80.0 mmol) of thionyl chloride to the suspension.
Add dropwise with mechanical stirring at such a rate as to maintain the reaction temperature at -5 to -10°C. The mixture is allowed to warm to room temperature, stirred overnight, and concentrated under reduced pressure. The off-white crystalline residue is treated with ethyl ether (twice) and dried under reduced pressure to give 6.25 g of 4-aminobutanoic acid methyl ester hydrochloride as an almost white solid. Melting point 118.5-122℃ (softens at 90℃). b (±)-4-[[2-[(acetylthio)methyl]
-1-oxo-3-phenylpropyl]amino]
Preparation of methyl butanoic acid ester: - Add 0.95 g (4.0 mmol) of (±)-2-[(acetylthio)methyl]-3-phenylpropanoic acid to 8 ml of ethyl ether under a nitrogen atmosphere. This mixture is carefully treated with a catalytic amount of dimethylformamide (3 drops) and stirred for 1 hour at room temperature. After concentration under reduced pressure, the residue is treated with 10 ml of tetrahydrofuran and concentrated again to remove excess oxalyl chloride. The resulting residue is treated with 5 ml of methylene chloride and the soluble portion is transferred to a dropping funnel. Add this solution to 4
-Aminobutanoic acid methyl ester hydrochloride 0.68g
(4.4 mmol) and diisopropylethylamine
1.6 ml (9.0 mmol) of methylene chloride 10 ml cold (−
5° C.) solution was added dropwise over 10 minutes with stirring under a nitrogen atmosphere. After cooling and stirring for 3 hours, the yellow solution is concentrated under reduced pressure. Dissolve the residue in 60ml of ethyl acetate.
to remove diisopropylethylamine hydrochloride, add 10% potassium hydrogen sulfate, water, 50% saturated sodium bicarbonate and 50% saline (30% each
ml). Dry the organic layer over sodium sulfate to obtain 1.34 g of a light brown solid. This solid is adsorbed onto a small amount of silica gel (230-400 mesh) and applied to a similar 80 g column of silica. Eluted with hexane/acetone (5:2) as a white solid (±)-4-[[2-[(acetylthio)methyl]-
1.09 g of 1-oxo-3-phenylpropyl]amino]butanoic acid methyl ester are obtained. Melting point 81-82
℃. c (±)-4-[[2-(mercaptomethyl)-1
- Preparation of oxo-3-phenylpropyl]amino]butanoic acid: - Under a nitrogen atmosphere, 0.6 g (1.8 mmol) of the product from section b above is dissolved in 5 ml of methanol and cooled on an ice bath. Add 5.4 ml of 1N sodium hydroxide to this solution (3
equivalent amount) over 10 minutes. The mixture is stirred in the cold for 3 hours and concentrated under reduced pressure to about 1/2 volume.
The residue was diluted with 40 ml of water and diluted with chloroform (15 ml x
Wash twice) and adjust the pH to approximately 1.5 with concentrated hydrochloric acid. The resulting white suspension is extracted with ethyl acetate (15 ml x 3). These extracts were combined, washed with water and brine (15 ml each), dried over sodium sulfate, concentrated and yielded (±)-4-[[2-
0.48 g of (mercaptomethyl)-1-oxo-3-phenylpropyl]amino]butanoic acid was obtained. melting point
94.5-101℃. R _f =0.41 for TLC (on silica gel, benzene/acetic acid (4:1)). Elemental analysis, calculated as C ₁₄ H ₁₉ NO ₃ S: C, 59.76%; H, 6.81%; N, 4.98
%; S, 11.40%; Actual value: C, 59.81%; H, 6.81%; N, 4.96
%; S, 11.37%. Example 3 (S)-3-[[2-(mercaptomethyl)-1-
Production of oxo-3-phenylpropyl]amino]benzenebutanoic acid (isomer A): - a Production of (S)-4-phenyl-3-[[(phenylmethoxy)carbonyl]amino]butanoic acid methyl ester :- Preparation of its mixed anhydride using isobutyl chloroformate in ethyl ether and the method of Penke et al. [Helvetica quimica acta (Helv.
(S)-[3-diazo-2-oxo-1-(phenylmethyl)propyl]carbamic acid phenylmethyl ester was obtained according to the method described in Chim Acta, Vol. 53, pp. 1057-1061 (1970). silver benzoate
1.0 g (4.7 mmol) is mixed with 10 ml of triethylamine and filtered. A portion of the liquid (approximately 0.5 ml) is added to a solution of 2.0 g (6.2 mmol) of the above diazo compound in 20 ml of methanol. Gas evolution occurs and the mixture becomes dark in color. After the effervescence had subsided (approximately 10 minutes), the mixture was further treated with silver benzoate solution (approximately 0.5 ml),
Stir for 30 minutes. The mixture is treated with activated charcoal, filtered and concentrated under reduced pressure to give a dark tar-like residue. Chromatography through Florisil (ethyl ether eluent) gave 1.65 g of (S)-4-phenyl-3-[[(phenylmethoxy)carbonyl]amino]butanoic acid methyl ester as a light yellow solid. get. b Production of (S)-3-amino-4-phenylbutanoic acid methyl ester/p-toluenesulfonate: - Under argon atmosphere, (S)-4-phenyl-3-
A solution of 1.64 g (5.0 mmol) of [(phenylmethoxy)carbonyl]amino]butanoic acid methyl ester and 0.95 g (1.0 equivalent) of p-toluenesulfonic acid hydrate in 30 ml of 95% ethanol was mixed with a 10% palladium/activated carbon catalyst. Treat with 100mg. The mixture was stirred under hydrogen pressure for 1.5 hours, filtered and concentrated to give an off-white residue. Recrystallized from acetonitrile (heated) to give (S)-3-amino-4-phenylbutanoic acid methyl ester p- as needle-like white crystals.
1.44 g of toluenesulfonate is obtained. Melting point 168.5
~170.5℃. [α] ²⁷ _D +7.0° (in methanol, C=
1.05). c (S)-3-[[2-[(acetylthio)methyl]
-1-oxo-3-phenylpropyl]amino]
Preparation of benzenebutanoic acid methyl ester (isomer A): - Under argon atmosphere, (±)-2-[(acetylthio)methyl]-3-phenylpropanonic acid 0.92 g
To a solution of (3.9 mmol) in 30 ml of ethyl ether is added oxalyl chloride (1.1 eq.). Carefully treat this solution with dimethylformamide (2 drops) and stir for 45 minutes. The mixture was concentrated under reduced pressure;
The residue is dissolved in 5 ml of methylene chloride. Under argon atmosphere, (S)-3-amino-4-phenylbutanoic acid methyl ester p-toluenesulfonate
1.41 g (3.84 mmol) and diisopropylethylamine 1.35 ml (7.75 mmol) of methylene chloride 20
ml cold (ice/methanol) solution to the above methylene chloride solution dropwise over 10 minutes. The mixture is warmed to room temperature and stirred for 3 hours, then concentrated. 50 residue
Partition between 100 ml of saturated sodium bicarbonate and ethyl acetate (3 x 30 ml). Combine the organic layers and add water.
10% potassium hydrogen sulfate and saline (50% each
ml), dried over sodium sulfate and concentrated,
1.57 g of a yellow oil (slowly crystallizing) is obtained. This oil is applied to a 160 g column of silica gel (230-400 mesh) (preadsorbed onto a small amount of similar silica) and eluted with 26% ethyl acetate/petroleum ether. Fractions containing only the faster migrating isomer were combined and concentrated to give (S)-3-[[2-[(acetylthio)methyl]-1 as a white crystalline solid.
-oxo-3-phenylpropyl]amino]benzenebutanoic acid methyl ester (isomer A) 0.702
get g. Melting point 87-88.5℃. [α] ²⁶ _D −77.5° (c=0.96 in chloroform). d (S)-3-[[2-(mercaptomethyl)-1
- Preparation of oxo-3-phenylpropyl]amino]benzenebutanoic acid (isomer A): - under argon atmosphere, 0.35 g of the product of section c above.
1.87 ml (2.2 equivalents) of 1N sodium hydroxide was added dropwise to a 2 ml methanol solution of (0.85 mmol) over 10 minutes. After 2 hours, a further 0.7 ml of 1N sodium hydroxide is introduced and the mixture is treated with sufficient methanol to dissolve the precipitate. After stirring for 7 hours at room temperature, the methanol is evaporated off under reduced pressure. The residue was diluted with water, treated with 2N sodium hydroxide to dissolve the solids, and treated with ethyl acetate (30 ml x
twice) and then extracted with hexane (30 ml). Adjust the pH of the water tank to approximately 1.0 with concentrated hydrochloric acid, and extract with ethyl acetate (20 ml x 3 times). The combined organic layers were washed with water and brine (30 ml each), dried over sodium sulfate and concentrated under reduced pressure to give (S)-3-[[2-(mercaptomethyl)-1-oxo] as a white solid. 0.23 g of -3-phenylpropyl]amino]benzenebutanoic acid (isomer A) was obtained. Melting point 149~
151.5℃. [α] ²⁶ _D +40.3° (in methanol, c=
1.00). TLC (on silica gel, benzene/acetic acid (7:1) R _f = 0.40. Elemental analysis, calculated as C ₂₀ H ₂₃ NO ₃ S: C, 67.20%; H, 6.49%; N, 3.92
%; S, 8.97%; Actual value: C, 66.86%; H, 6.45%; N, 3.93
%; S, 8.71%. Example 4 (S)-3-[[2-(mercaptomethyl)-1-
Production of oxo-3-phenylpropyl]amino]benzenebutanoic acid (isomer B): - a (S)-3-[[2-[(acetylthio)methyl]
-1-oxo-3-phenylpropyl]amino]
Preparation of benzene butanoic acid methyl ester (isomer B):- The fractions containing the slower migrating isomer from Example 3c above are combined and concentrated and treated according to Example 3c as a white solid. 0.67 g of (S)-3-[[2-[(acetylthio)methyl]-1-oxo-3-phenylpropyl]amino]benzenebutanoic acid methyl ester (isomer B) is obtained. Melting point 93.5-95℃.
[α] ²⁶ _D +55.4° (in chloroform, c=1.00). b (S)-3-[[2-(mercaptomethyl)-1
- Preparation of oxo-3-phenylpropyl]amino]benzenebutanoic acid (isomer B): - under argon atmosphere, 0.35 g of the product of section a above.
A solution of 0.102 g (3 equivalents) of sodium hydroxide in 1.5 ml of a 20% aqueous methanol solution was added dropwise over 15 minutes to a 3 ml methanol solution of (0.85 mmol) with stirring. After stirring for 6 hours at room temperature, the reaction was incomplete (TLC), so an additional amount of sodium hydroxide (0.85 ml of 1N aqueous sodium hydroxide solution) was introduced into the mixture and kept in a cold place (refrigerator) overnight. . The next day, methanol was removed under reduced pressure and
The residue is partitioned between 40 ml of water and ethyl acetate (2 x 25 ml). Adjust the pH of the aqueous layer to approximately 1.0 with concentrated hydrochloric acid, and extract with ethyl acetate (20 ml x 3). These extracts were combined, washed with water and brine (30 ml each), dried over sodium sulfate and concentrated under reduced pressure.
0.30 g of (S)-3-[[2-(mercaptomethyl)-1-oxo-3-phenylpropyl]amino]benzenebutanoic acid (isomer B) was obtained as a white solid. Melting point 148-150℃. [α] ²⁶ _D −28.9° (in methanol, c=1.07). TLC (on silica gel, benzene/
R _f of acetic acid (7:1) = 0.34. Elemental analysis, as C ₂₀ H ₂₃ NO ₃ S, calculated values: C, 67.20%; H, 6.49%; N, 3.92
%; S, 8.97%; Actual value: C, 66.95%; H, 6.53%; N, 3.82
%; S, 8.82%. Example 5 (±)-5-[[2-(mercaptomethyl)-1-
Preparation of oxo-3-phenylpropyl]amino]pentanoic acid:-a Preparation of 5-aminovaleric acid methyl ester hydrochloride:-2.34 g of 5-aminovaleric acid (20
2.93 ml of thionyl chloride in a cold suspension of
(40 mmol) is added dropwise with stirring at such a rate that the reaction temperature is maintained between -5 and -10°C. After the dropwise addition of thionyl chloride is complete, the mixture is warmed to room temperature and stirred overnight. The mixture was concentrated under reduced pressure and the resulting white crystals were treated twice with ether to give 5-aminovaleric acid methyl ester hydrochloride as a white solid.
Obtain 2.68g. Melting point 132-137℃ (softens at 86℃). b (±)-5-[[2-[(acetylthio)methyl]
-1-oxo-3-phenylpropyl]amino]
Preparation of pentanoic acid methyl ester: - (±)-2-[(acetylthio)methyl]-3-phenylpropanoic acid (1.19 g (5 mmol)) in 10 ml of ethyl ether solution with 0.45 ml of oxalyl chloride
(5.2 mmol) is added. The mixture is carefully treated with a catalytic amount of dimethylformamide (3 drops) and stirred for 1 hour at room temperature. The mixture is concentrated under reduced pressure, the oily product is dissolved in 10 ml of tetrahydrofuran and concentrated again under reduced pressure. The residue was dissolved in 6 ml of methylene chloride and mixed with 0.91 g (5.4 mmol) of 5-aminovaleric acid methyl ester hydrochloride and 1.96 ml (11.25 mmol) of diisopropylethylamine.
Add dropwise to 11 ml of cold (-5°C) solution of methylene chloride over 10 minutes with stirring. In a cold place (-5℃)
After stirring for 2.5 hours, the mixture is allowed to rise to room temperature and stirred overnight. The next day the mixture is concentrated under reduced pressure and the residue is taken up in 60 ml of ethyl acetate to remove diisopropylethylamine hydrochloride. 10 liquid
% potassium hydrogen sulfate, water, 50% saturated sodium hydrogen carbonate and 50% saline (20 ml each x 3 times)
Wash sequentially with The organic layer is dried over sodium sulfate and concentrated to give 1.49 g of a very light yellow oil. This oil was applied to a 115 g column of silica gel (230-400 mesh) and eluted with hexane/acetone (5:2) as an anhydrous oil with (±) -5
1.41 g of -[[2-[(acetylthio)methyl]-1-oxo-3-phenylpropyl]amino]pentanoic acid methyl ester is obtained. c (±)-5-[[2-(mercaptomethyl)-1
-Production of oxo-3-phenylpropyl]amino]pentanoic acid:- Under nitrogen atmosphere, 1.20 g (3.41
mmol) in 13 ml of methanol cold solution (ice bath),
12.9 ml (3.8 equivalents) of 1N sodium hydroxide are added dropwise over 10 minutes. The mixture is stirred at 0° C. for 10 minutes, allowed to rise to room temperature, and stirred for 3 hours. The mixture is concentrated to half volume under reduced pressure. The residue was diluted with 40 ml of water and diluted with chloroform (20 ml x
Wash twice) and adjust the pH to approximately 1.5 with concentrated hydrochloric acid. The resulting white suspension is extracted with ethyl acetate (15 ml x 3). The combined organic extracts are washed with water and brine (20 ml each), dried over sodium sulfate, and concentrated to give 0.82 g of a white wax-like solid. Dissolve the solid in ethyl acetate, filter, concentrate,
The residue was treated with ether as a white solid (±)
-5-[[2-(mercaptomethyl)-1-oxo-
3-phenylpropyl]amino]pentanoic acid was obtained. Melting point 78-79℃. TLC (on silica gel, benzene/acetic acid (4:1) R _f = 0.30 (slight tailing observed). Elemental analysis, calculated as C ₁₅ H ₂₁ NO ₃ S: C, 60.99%; H, 7.17 %;N, 4.74
%; S, 10.85%; Actual value: C, 60.80%; H, 7.15%; N, 4.62
%; S, 10.73%. Example 6 (±)-6-[[2-(mercaptomethyl)-1-
Preparation of oxo-3-phenylpropyl]amino]hexanoic acid: a. Preparation of 6-aminocaproic acid methyl ester hydrochloride: - 2.62 g of 6-aminocaproic acid in 25 ml of methanol.
(20 mmol) of thionyl chloride 2.93
ml (40 mmol) are added dropwise with stirring at such a rate that the reaction temperature is maintained between -5 and -10°C. After the thionyl chloride addition is complete, the mixture is allowed to warm to room temperature and stirred for 4 hours. The mixture is concentrated under reduced pressure and the resulting white solid is treated twice with ether to obtain 3.53 g of 6-aminocaproic acid methyl ester hydrochloride as a white solid. Melting point 114-118℃
(Softens above 109℃). b (±)-6-[[2-[(acetylthio)methyl]
-1-oxo-3-phenylpropyl]amino]
Production of hexanoic acid methyl ester: - (±)-2-[(acetylthio)methyl]-3-phenylpropanoic acid (1.19 g (5 mmol)) in a solution of 10 ml of ethyl ether (0.45 ml of oxalyl chloride)
(5.2 mmol) is added. The mixture is carefully treated with a catalytic amount of dimethylformamide (3 drops) and stirred for 1 hour at room temperature. The mixture was concentrated under reduced pressure and the resulting oil was dissolved in 10 ml of tetrahydrofuran.
and concentrated again under reduced pressure. Dissolve the residue in 6 ml of methylene chloride and add 0.98 g (5.4 mmol) of 6-aminocaproic acid methyl ester hydrochloride.
and 1.96 ml (11.25 mmol) of diisopropylethylamine in 11 ml of cold (-5°C) methylene chloride solution with stirring over 10 minutes. After stirring for 2.5 hours in the cold (-5°C), the mixture is allowed to warm to room temperature and stirred for 2 hours. The mixture is concentrated under reduced pressure and the residue is taken up in 60 ml of ethyl acetate to remove diisopropylethylamine hydrochloride. The solution was diluted with 10% potassium bisulfate, water, 50% saturated sodium bicarbonate and 50% saline (each
Wash sequentially with 20 ml x 3 times). The organic layer was dried over sodium sulfate and concentrated to give methyl (±)-6-[[2-[(acetylthio)methyl]-1-oxo-3-phenylpropyl]amino]hexanoate as a slightly yellow solid. 1.64 g of ester is obtained.
Melting point 50-54℃ (sintered above 39℃). c (±)-6-[[2-(mercaptomethyl)-1
-Production of oxo-3-phenylpropyl]amino]hexanoic acid:- Under nitrogen atmosphere, 1.46 g (3.99 g) of the product of item b above
mmol) in methanol (12 ml cold solution (ice bath)),
Add 12 ml (3 equivalents) of 1N sodium hydroxide dropwise over 10 minutes. The mixture is stirred at 0° C. for 10 minutes, allowed to warm to room temperature, and stirred for 3 hours.
The mixture is concentrated to half volume under reduced pressure. Water the residue
Dilute with 40 ml, wash with chloroform (2 x 20 ml), and adjust the aqueous portion to pH approximately 1.5 with concentrated hydrochloric acid. The resulting white suspension was diluted with ethyl acetate (15 ml x 3 times).
Extract. The combined extracts were washed with water and brine (20 ml each), dried over sodium sulfate, and concentrated to give 1.23 g of a colorless oil. This oil is applied to an 80 g column of silica gel (230-400 mesh) and eluted with 20% acetic acid/toluene to yield 0.99 g of a colorless oil. The oil was chromatographed again on silica (230-400 mesh).
Elution with 15% acetic acid/toluene gives 0.75 g of colorless oil.
get. This material is applied to a Silic AR.cc-4 silica column and eluted with chloroform with gradual addition of ethyl acetate (10-40% ethyl acetate) to yield 0.67 g of a white solid. This solid was treated with hexane/ether as a white solid (±)-6-[[2-
0.456 g of (mercaptomethyl)-1-oxo-3-phenylpropyl]amino]hexanoic acid was obtained.
Melting point 58-60℃. TLC (on silica, toluene/acetic acid (4:1)) R _f =0.26 (slight tailing observed). Elemental analysis, as _C16H23NO3S・_0.18H2O , calculated values: C, 61.46%; _H , 7.53%; _N , 4.48
%; S, 10.25%; Actual value: C, 61.46%; H, 7.47%; N, 4.40
%; S, 10.14%. Example 7 (±)-5-[[2-(mercaptomethyl)-1-
Production of oxo-3-phenylpropyl]amino]pentanoic acid methyl ester: - Under nitrogen atmosphere, (±)-5-[[2-[(acetylthio)methyl]-1-oxo-3-phenylpropyl]amino ] Pentanoic acid methyl ester 1.93g
(5.5 mmol) in 10 ml of methanol and cool in an ice bath. Add 5.5ml of 1N sodium hydroxide to this solution.
(1.0 eq.) was added dropwise over 10 minutes. The mixture is stirred at 0° C. for 10 minutes, allowed to warm to room temperature, and stirred for 1 hour. The mixture is concentrated to half volume under reduced pressure, the residue is diluted with 20 ml of water and extracted with ethyl acetate (3 x 15 ml). The combined organic layers were washed with water and brine (15 ml each), dried over sodium sulfate, and concentrated to give 1.68 g of a colorless oil.
Merck silica gel (230 ~
Approximately 125 g (240 mesh) was applied to a column and eluted with hexane/acetone (5:3) to give (±)-5-[[2-(mercaptomethyl)-1-] as a white solid.
1.34 g of oxo-3-phenylpropyl]amino]pentanoic acid methyl ester was obtained. Melting point 39-41
℃. TLC (on silica gel, hexane/acetone (5:3)) R _f =0.29 (slight tailing observed). Elemental analysis, calculated as C ₁₆ H ₂₃ NO ₃ S: C, 62.11%; H, 7.49%; N, 4.53
%; S, 10.36%; SH, 10.69%; Actual value: C, 61.86%; H, 7.54%; N, 4.50
%; S, 10.21%; SH, 10.62%. Example 8 (±)-7-[[2-[(acetylthio)methyl]-
Preparation of 1-oxo-3-phenylpropyl]amino]heptanoic acid methyl ester:-a Preparation of 7-aminoheptanoic acid methyl ester hydrochloride (1:1):-1.8 g of 7-aminoheptanoic acid in 30 ml of methanol.
(12.4 mmol) of thionyl chloride in a cold suspension of 1.81
ml (24.8 mmol) at a reaction temperature of -5 to -10℃
Add dropwise with stirring at a rate such that . After adding all of the thionyl chloride dropwise, the mixture is allowed to warm to room temperature and stirred overnight. The mixture is concentrated under reduced pressure and the resulting white solid is treated twice with ether to obtain 2.38 g of 7-aminoheptanoic acid methyl ester hydrochloride (1:1) as a white solid. melting point
132-134℃ (sintered above 118℃). b (±)-7-[[2-[(acetylthio)methyl]
-1-oxo-3-phenylpropyl]amino]
Production of heptanoic acid methyl ester: - 3-acetylthio-2-benzylpropanoic acid
1.83 g (7.67 mmol) of ethyl ether (15 ml
In solution, 0.67 ml (7.69 mmol) of oxalyl chloride
Add. The mixture is carefully treated with a catalytic amount of dimethylformamide (2 drops) and stirred for 1 hour at room temperature. The mixture is concentrated under reduced pressure, the oil obtained is dissolved in 15 ml of tetrahydrofuran and concentrated again under reduced pressure. The residue was dissolved in 15 ml of methylene chloride and mixed with 1.50 g (7.67 mmol) of 7-aminoheptanoic acid methyl ester hydrochloride (1:1) and 2.68 ml (15.38 mmol) of diisopropylethylamine in 15 ml of cold methylene chloride (-5°C). ) in solution, 10
Add dropwise with stirring over a period of minutes. Cold place (15
After stirring for 2.5 hours at ), the mixture is allowed to warm to room temperature and stirred overnight. The mixture is concentrated under reduced pressure and the residue is taken up in 100 ml of ethyl acetate to remove diisopropylethylamine hydrochloride. liquid
10% potassium hydrogen sulfate, water, 5% saturated sodium hydrogen carbonate, water and 50% saline (30 ml each
3 times). The organic layer was dried over sodium sulfate and concentrated to obtain 2.60 g of a white solid. This solid was recrystallized from hexane/ethyl acetate as a white solid (±)-7-[[2-[(acetylthio)methyl]-1-oxo-3-phenylpropyl]amino]heptanoic acid methyl ester 2.48 g I got it.
Melting point 84-85℃ (sintered above 51℃). R _f of TLC (on silica gel, ethyl acetate/hexane (2:1))
=0.45. Elemental analysis, calculated as C ₂₀ H ₂₉ NO ₄ S: C, 63.30%; H, 7.70%; N, 3.69
%; S, 8.45%; Actual value: C, 63.01%; H, 7.57%; N, 3.51
%; S, 8.40%. Example 9 (±)-7-[[2-(mercaptomethyl)-1-
Production of oxo-3-phenylpropyl]amino]heptanoic acid: - (±) -7-[[2-[acetylthio)methyl]-
1-oxo-3-phenylpropyl]amino]heptanoic acid methyl ester 1.65g (4.3mmol)
Dissolve in 20 ml of methanol by heating and cool in an ice bath under nitrogen atmosphere. To this solution, 13 ml (approximately 3 equivalents) of 1N sodium hydroxide is added dropwise over 10 minutes. The mixture is stirred at 0° C. for 10 minutes, then allowed to warm to room temperature and stirred for 3 hours. The mixture is concentrated under reduced pressure to remove all methanol. The residue was diluted with 40 ml of water and diluted with chloroform (20 ml x 2
3 times) Wash and adjust the aqueous part to pH approximately 1.5 with concentrated hydrochloric acid. The resulting white suspension was diluted with ethyl acetate (15 ml
3 times) Extract. The combined extracts were washed with water and brine (20 ml each), dried over sodium sulfate, and concentrated to give 1.32 g of a white solid. About 90 g of solid Merck silica gel (230-400 mesh)
Apply to the column and elute with toluene/acetic acid (7:1) to obtain 1.14 g of a white solid. This material was applied to a silica gel (Silic AR.CC-4) column, and chloroform and ethyl acetate (10-55%
(±)-7-[[2
1.05 g of -(mercaptomethyl)-1-oxo-3-phenylpropyl]amino]heptanoic acid was obtained.
Melting point 71-73℃ (sintered above 65℃). TLC (on silica gel, benzene/acetic acid (4:1) R _f =0.36
(trace in 0.27). Elemental analysis, calculated as C ₁₇ H ₂₅ NO ₃ S: C, 63.13%; H, 7.79%; N, 4.30
%; S, 9.91%; SH, 10.22%; Actual value: C, 63.36%; H, 7.91%; N, 4.21
%; S, 9.57%; SH, 9.89%. Example 10 (±)-7-[[2-(mercaptomethyl)-1-
Production of oxo-3-phenylpropyl]amino]heptanoic acid methyl ester:- (±)-7-[[2-[(acetylthio)methyl]-
1-oxo-3-phenylpropyl]amino]heptanoic acid methyl ester 1.43g (3.77mmol)
Dissolve in 30 ml of methanol with heating and cool on an ice bath under nitrogen atmosphere. 3.77 ml (1.0 equivalent) of 1N sodium hydroxide is added dropwise to this solution over 10 minutes. The mixture is stirred at 0° C. for 10 minutes and then allowed to warm to room temperature for 1 hour. The mixture is concentrated under reduced pressure to remove all methanol. The residue was diluted with 20 ml of water and diluted with ethyl acetate (15
ml x 3 times). The combined organic layers were washed with water and brine (15 ml each), dried over sodium sulfate, and concentrated to give 1.26 g of a white solid.
The solid is dissolved in 25 ml of a mixture of methanol:water (1:1) and the pH is adjusted to approximately 1.0 with concentrated hydrochloric acid. Add 0.080 g of zinc dust to this solution and stir the mixture for 30 minutes at room temperature. Remove residual zinc powder, remove all methanol from the liquid, and concentrate. The residue is diluted with 20 ml of water and extracted with ethyl acetate (3 x 25 ml). The combined organic layers were washed with 5% sodium bicarbonate, water, and brine (2 x 20 ml each), dried over sodium sulfate, and concentrated to yield 1.23 g of a white solid. This material was purified by flash chromatography on 50 g of Merck silica gel (230-400 mesh) and eluting with ethyl acetate to give 0.980 g of a white solid. The solid was dissolved in methanol, filtered through a cellulose microfilter, and concentrated under reduced pressure to give (±)-7-[[2-(mercaptomethyl)-1-oxo-3-phenylpropyl]amino]heptane. Acid methyl ester was obtained. Melting point 68-70℃. R _f =0.44 for TLC (on silica gel, benzene/acetic acid (4:1)). Elemental analysis, calculated as C ₁₈ H ₂₇ NO ₃ S: C, 64.06%; H, 8.06%; N, 4.15
%; S, 9.50%; SH, 9.80%; Actual value: C, 64.33%; H, 8.14%; N, 3.99
%; S, 9.45%; SH, 9.79%. Example 11 (±)-8-[[2-(mercaptomethyl)-1-
Preparation of oxo-3-phenylpropyl]amino]octanoic acid:-a Preparation of 8-aminoactanoic acid methyl ester hydrochloride (1:1):-3.18 g of 8-aminooctanoic acid in 25 ml of methanol.
(20 mmol) of thionyl chloride in a cold suspension of 2.93 ml
(40 mmol) is added dropwise with stirring at such a rate that the reaction temperature is maintained between -5 and -10°C. After adding all of the thionyl chloride dropwise, the mixture is allowed to warm to room temperature and stirred overnight. The mixture was concentrated under reduced pressure and the resulting white solid was treated twice with ether,
3.98 g of 8-aminooctanoic acid methyl ester hydrochloride (1:1) are obtained as a white solid. Melting point 130~
132℃. b (±)-8-[[2-[(acetylthio)methyl]
-1-oxo-3-phenylpropyl]amino]
Production of octanoic acid methyl ester: - 3-acetylthio-2-benzylpropanoic acid
To a solution of 1.19 g (5 mmol) in 10 ml of ethyl ether is added 0.45 ml (5.2 mmol) of oxalyl chloride. The mixture is carefully treated with a catalytic amount of dimethylformamide (3 drops) and stirred for 1 hour at room temperature. The mixture is concentrated under reduced pressure and the resulting oil is dissolved in 10 ml of tetrahydrofuran and concentrated again under reduced pressure. This residue was dissolved in 6 ml of methylene chloride and mixed with 1.06 g (5.1 mmol) of 8-aminooctanoic acid methyl ester hydrochloride (1:1) and 1.96 ml (11.25 mmol) of diisopropylethylamine in 11 ml of cold (-5 °C) solution, 10
Add dropwise with stirring over a period of minutes. Cold place (-5
After stirring for 2.5 hours at ), allow the mixture to warm to room temperature and stir overnight. The mixture is concentrated under reduced pressure and the residue is taken up in 60 ml of ethyl acetate to remove diisopropylethylamine hydrochloride. The solution was mixed with 10% potassium hydrogen sulfate, water, 5% saturated sodium hydrogen carbonate, water and 50% saline (20 ml each).
3 times). The organic layer was dried over sodium sulfate and concentrated to give 2.01 g of a light yellow solid. Dissolve the solid in ethyl acetate and add Merck silica (230~
400 mesh) Adsorb a small amount onto the top. This material was applied to a similar silica ca.
-1-oxo-3-phenylpropyl]amino]
1.69 g of octanoic acid methyl ester are obtained. melting point 44
~45℃. c (±)-8-[[2-(mercaptomethyl)-1
- Preparation of oxo-3-phenylpropyl]amino]octanoic acid: - Under a nitrogen atmosphere, 1.69 g (4.3 mmol) of the methyl ester product from section b above are dissolved in 13 ml of methanol and cooled on an ice bath. To this solution, 13 ml (approximately 3 equivalents) of 1N sodium hydroxide is added dropwise over 10 minutes. The mixture is stirred at 0° C. for 10 minutes, allowed to warm to room temperature, and stirred for 4 hours. Flow argon into this solution and store it in the refrigerator overnight (approx.
After stirring for 3.5 hours, a white precipitate forms). Methanol is added and the solution is concentrated under reduced pressure to remove all methanol. The residue was diluted with 40 ml of water, washed with chloroform (2 x 20 ml), and the aqueous portion was washed with concentrated hydrochloric acid.
Adjust the pH to approximately 1.5. The resulting white suspension is extracted with ethyl acetate (15 ml x 3). These extracts were combined, washed with water and brine (20 ml each), dried over sodium sulfate, and concentrated to a white solid.
Obtain 0.720g. Re-extract the first chloroform layer with 1N sodium hydroxide (2 x 15 ml). The aqueous layer is acidified, combined with the previously acidified layer and extracted with ethyl acetate (2 x 20 ml). These extracts are washed with water and brine, dried, and concentrated to yield an additional 0.380 g of white solid. Combine the two parts of the solid (±)
-8-[[2-(mercaptomethyl)-1-oxo-
3-phenylpropyl]amino]octanoic acid 1.10
I got g. Melting point 81-83℃ (sintered above 68℃).
TLC (on silica gel, benzene/acetic acid (4:1))
R _f =0.42. Elemental analysis, calculated as C ₁₈ H ₂₇ NO ₃ S: C, 64.06%; H, 8.06%; N, 4.15
%; S, 9.50%; SH, 9.80%; Actual value: C, 63.77%; H, 7.84%; N, 4.12
%; S, 9.18%; SH, 9.66%. Example 12 (±)-11-[[2-(mercaptomethyl)-1-
Preparation of oxo-3-phenylpropyl]amino]undecanoic acid:-a Preparation of 11-aminoundecanoic acid methyl ester hydrochloride (1:1):-8.05 11-aminoundecanoic acid in 50 ml of methanol
g (40 mmol) of thionyl chloride in a cold suspension of 5.84
ml (80 mmol) are added dropwise with stirring at such a rate that the reaction temperature is maintained between -5 and -10°C.
After adding all of the thionyl chloride dropwise, the mixture is allowed to warm to room temperature. Add 50ml of methanol to this mixture.
Dissolve the white precipitated product by adding and stir overnight at room temperature. The mixture was concentrated under reduced pressure and the resulting white solid was treated twice with ether to give 11-aminoundecanoic acid methyl ester hydrochloride (1:
1) Obtain 7.26g. Melting point 155-158℃. b (±)-11-[[2-[(acetylthio)methyl]
-1-oxo-3-phenylpropyl]amino]
Production of undecanoic acid methyl ester:-3-acetylthio-2-benzylpropanoic acid
To a solution of 1.43 g (6 mmol) in 10 ml of ethyl ether is added 0.54 ml (6.2 mmol) of oxalyl chloride. The mixture is carefully treated with a catalytic amount of dimethylformamide (3 drops) and stirred for 1 hour at room temperature. The mixture is concentrated under reduced pressure and the resulting oil is dissolved in 10 ml of tetrahydrofuran and concentrated again under reduced pressure. This residue was dissolved in 6 ml of methylene chloride and mixed with 1.61 g (6.4 mmol) of 11-aminoundecanoic acid methyl ester hydrochloride (1:1) and 2.23 ml (12.8 mmol) of diisopropylethylamine in 11 ml of cold (-5 °C) solution, 10
Add dropwise with stirring over a period of minutes. Cold place (-5
After stirring for 2.5 hours at ), allow the mixture to warm to room temperature and stir overnight. The mixture is concentrated under reduced pressure and the residue is taken up in 100 ml of acetic acid to remove diisopropylethylamine hydrochloride. Add the solution to 10% potassium hydrogen sulfate, water, 5% sodium hydrogen carbonate,
Wash sequentially with water and 50% saline (3x30ml each). The organic layer was dried over sodium sulfate and concentrated to give 2.5 g of (±)-11-[[2-[(acetylthio)methyl]-1-oxo-3-phenylpropyl]amino]undecanoic acid methyl ester as a light yellow solid. get. Melting point 64-66℃. c (±)-11-[[2-(mercaptomethyl)-1
- Preparation of oxo-3-phenylpropyl]amino]undecanoic acid: - Under a nitrogen atmosphere, 2.5 g (5.74 mmol) of the methyl ester product from section b above are dissolved in 25 ml of methanol and cooled in an ice bath. Add 23 ml (approximately 3 equivalents) of 1N sodium hydroxide to this solution dropwise over 10 minutes (the solution becomes cloudy after the entire amount of sodium hydroxide is added dropwise).
The mixture is stirred at 0° C. for 10 minutes, allowed to warm to room temperature, and stirred for 3 hours. The mixture is concentrated under reduced pressure to remove all methanol. Pour the residue into 40ml of water
and extract with chloroform (2x20ml). Extract the chloroform layer with 1N sodium hydroxide (2x20ml). Combine all the aqueous extracts and adjust the pH to approximately 1.5 with concentrated hydrochloric acid. The white suspension formed is extracted with ethyl acetate (40 ml x 3). These extracts were combined and dissolved in water and saline (respectively).
40 ml), dried over sodium sulfate and concentrated to give 2.09 g of a light yellow solid. This solid is hexane,
Recrystallization from a mixture of ethyl acetate and ether gives impure crystals. Therefore, this material was dissolved in ethyl acetate and silica gel (Silic AR.CC−
4) Adsorb onto a small volume and apply to a column made of similar silica. Elute with chloroform and ethyl acetate (10-40% ethyl acetate) as a white solid (±)−11
-[[2-(Mercaptomethyl)-1-oxo-3-
Phenylpropyl]amino]undecanoic acid 1.77g
I got it. Melting point 104-105℃. TLC (on silica gel,
R _f =0/51 for benzene/acetic acid (4:1). Elemental analysis, calculated as C ₂₁ H ₃₃ NO ₃ S: C, 66.45%; H, 8.76%; N, 3.69
%; S, 8.45%; SH, 8.71%; Actual value: C, 66.12%; H, 8.75%; N, 3.65
%; S, 8.45%; SH, 8.67%. Example 13 (±)-5-[[2-(mercaptomethyl)-1-
Production of oxo-3-phenylpropyl]amino]pentamide: - a Production of 5-[[(1,1-dimethylethoxy)carbonyl]amino]pentanoic acid: - 2.93 g (25 mmol) of 5-aminopentanoic acid. To a stirred 7.5 ml solution of t-butanol:water (2:1) is added 1.0 g (25 mmol) of sodium hydroxide. Furthermore, t-butanol:water (2:
1) Add 9 ml to dissolve, then add 6.0 g (27.5 mmol) of di-t-butyl dicarbonate in several portions over 1 hour. Add another 5 ml of t-butanol and stir the thick white slurry for 3.5 days. The reaction mixture is diluted with 15 ml of water and extracted with hexane (3 x 15 ml). Acidify the aqueous layer with 3.5 g of potassium hydrogen sulfate (25 ml of water). The product was extracted with ethyl acetate (1 x 25 ml, 2 x 15 ml), washed with water and brine, dried over sodium sulfate and evaporated to give a colorless oil (slowly crystallizing)5.38
get g. This is treated with hexane-ethyl ether to obtain 4.29 g of 5-[[(1,1-dimethylethoxy)carbonyl]amino]heptanoic acid as a white crystalline solid. Melting point 47-50.5℃. b Preparation of 5-[[(1,1-dimethylethoxy)carbonyl]amino]pentamide: - 2.61 g (12 mmol) of 5-[[(1,1-dimethylethoxy)carbonyl]amino]pentanoic acid and N-methyl Add 1.55 ml (12 mmol) of isobutyl chloroformate to a solution of 1.32 ml (12 mmol) of morpholine in 15 ml of tetrahydrofuran (-15°C), stirring vigorously and keeping the reaction temperature at -25 to -15°C for about 5 minutes.
Drip over a period of minutes. Stir the mixture for 12 minutes,
10 ml of ammonia in methanol (saturated at 0°C)
is added dropwise over a period of 30 minutes at approximately -15°C. The mixture is stirred in the cold for an additional 30 minutes, allowed to warm to room temperature, and stirred overnight. The thick white suspension is diluted with water, concentrated under reduced pressure to remove the organic solvent and extracted with 150 ml of ethyl acetate. Ethyl acetate solution in water, 10%
Potassium hydrogen sulfate, washed with water and brine, dried over sodium sulfate, and evaporated to a white solid product.
Obtain 1.54g. Melting point 135-138.5℃. Due to low recovery, the initial aqueous extract was concentrated to half volume and diluted with ethyl acetate (1 x 75 ml, 3 x 25 ml).
times) extract. Add the ethyl acetate solution to water (10ml x 2
Wash with saline (10 ml x 2), dry with sodium sulfate, concentrate and 0.56 g of white solid (total amount)
2.1 g of product) are obtained. A small amount of the total product was recrystallized from ethyl acetate to give 5-[[(1,
Pure 1-dimethylethoxy)carbonyl]amino]pentamide is obtained. Melting point 138-140℃. c Preparation of 5-aminopentamide hydrochloride (1:1): - 1.77 g (8.18 mmol) of 5-[[(1,1-dimethylethoxy)carbonyl]amino]pentamide
is dissolved in 20 ml of 1.3N hydrogen chloride in acetic acid and the solution is left at room temperature for 45 minutes. The solvent is evaporated under reduced pressure and the oily residue is treated with cold ethyl ether to give a hard gum. Approximately 2 methanol in the product
ml of 5-aminopentamide hydrochloride (1:1) was added and triturated, and the resulting white fine powder was collected.
get g. d (±)-5-[[2-[(acetylthio)methyl]
-1-oxo-3-phenylpropyl]amino]
Production of pentamide: - 3-acetylthio-2-benzylpropanoic acid
1.43 g (6 mmol) and 0.58 ml of oxalyl chloride
(6.6 mmol) in 12 ml of ethyl ether, add 1 drop of dimethylformamide. The mixture is stirred at room temperature for 1 hour and concentrated under reduced pressure. 2 residues
Dissolve in portions in 5-10 ml of tetrahydrofuran, evaporate off the final traces of oxalyl chloride and reduce pressure with a vacuum pump for 10 minutes. A solution of this acid chloride in 7.5 ml of methylene chloride was added to 0.916 g (6 mmol) of 5-aminopentamide hydrochloride (1:1).
The mixture was added dropwise to a solution of 2.09 ml (12 mmol) of isopropylethylamine in 15 ml of methylene chloride at 5 to 10°C over 1 hour. The mixture is stirred in the cold for 1 hour, allowed to warm to room temperature, and stirred for 2 hours.
The solvent was evaporated and the residue was dissolved in 200 ml of ethyl acetate, water, 5% sodium bicarbonate, water, 10% potassium hydrogen sulfate, water and saline (approximately 20 ml each).
x2), dried over sodium sulfate, and concentrated under reduced pressure to obtain 1.44 g of a white solid. Recrystallized twice from ethyl acetate-hexane to give (±)-5
-[[2-[(acetylthio)methyl]-1-oxo-3-phenylpropyl]amino]pentamide
Obtain 0.86g. e (±)-5-[[2-(mercaptomethyl)-1
-Production of oxo-3-phenylpropyl]amino]pentamide:- (±)-5-[[2-[(acetylthio)methyl]-
0.85 g (2.53 mmol) of 1-oxo-3-phenylpropyl]amino]pentamide in methanol 5
ml solution (0-5℃), 1N sodium hydroxide
Add 2.53 ml (1 equivalent) dropwise over 5 minutes.
Remove the ice bath and stir the mixture for 1.5 hours. Most of the methanol is evaporated, the residue is diluted with about 7 ml of water and the pH is adjusted to 7.0 with 1N hydrochloric acid. The separated solid is extracted with ethyl acetate (10 ml, then 3 x 20 ml). The combined organic extracts are washed with brine, dried over sodium sulfate and concentrated under reduced pressure to yield 0.75 g of a white solid residue. Melting point 112-122℃. Methanol-
One recrystallization from water gives a material with a melting point of 122.5-125°C. This solid and mother liquor were combined again, concentrated, and chloroform-packed silica gel (Silic AR.CC) was used.
-4) Attach to column. The column was initially 10% ethyl acetate in chloroform, and increasing eluent polarity to ethyl acetate and finally 10% ethyl acetate.
Elution with a methanol eluent system yielded (±)-5-[[2-(mercaptomethyl)-1-oxo-3-phenylpropyl]amino]pentamide as a white solid.
0.50g was obtained. Melting point 127-128.5℃. TLC (on silica gel, benzene/acetic acid (7:3)) R _f =0.25
(weak tailing). Elemental analysis, calculated as C ₁₅ H ₂₂ N ₂ O ₂ S: C, 61.19%; H, 7.53%; N, 9.51
%; S, 10.89%; SH, 11.23%; Actual value: C, 60.88%; H, 7.64%; N, 9.28
%; S, 10.83%; SH, 11.11%. Examples 14 to 45 Using the amino compounds shown in the following columns and the carboxylic acids shown in the columns, the same treatments as in Examples 1 to 13 were carried out to obtain the acylmercaptoalkanoyl compounds shown in the respective columns. Furthermore, this compound was hydrolyzed to obtain the mercaptoalkanoyl compound shown in the column (the symbols in each formula are shown in the table).

【table】

【table】

【table】

【table】

[Table] In the cases of Examples 14-35, 38 and 43-45, hydrolysis of the acylmercaptoalkanoyl ester indicated in the column results in the corresponding mercaptoalkanoylcarboxylic acid (i.e. the compound in which R ₄ is hydroxy) or the corresponding mercaptoalkanoyl It is effective depending on the conditions in which one of the carboxylic acid esters (that is, the esters in which R ₄ is the ester group shown in the column (table)) is used. The hydroxy protecting group of Example 33 and the amino protecting group shown in Example 45 can be removed following completion of the reaction. Example 46 (±)-5-[[2-(mercaptomethyl)-1-
Production of oxo-3-phenylpropyl]amino]pentanoic acid sodium salt: - (±)-5-[[2-(mercaptomethyl)-1-
Oxo-3-phenylpropyl]amino]pentanoic acid (1 mmol) is dissolved in 50 ml of water. Add 20 ml of a 0.1N aqueous solution of sodium hydrogen carbonate to this, and freeze-dry this aqueous solution. Dissolve this in 10ml of water and use Sephadex Chromatography Egel G-
Apply to 10 columns (5 cm x 60 cm) and elute with water. Fractions containing the desired product were combined and lyophilized to yield (±)-5-[[2-(mercaptomethyl)-1-
Oxo-3-phenylpropyl]amino]pentanoic acid sodium salt was obtained. Example 47 From the following ingredients, (±) -5- per tablet
1000 tablets containing 100 mg of [[2-(mercaptomethyl)-1-oxo-3-phenylpropyl]amino]pentanoic acid are prepared according to the following manufacturing method. Component: (±)-5-[[2-(mercaptomethyl)-1-
Oxo-3-phenylpropyl]amino]pentanoic acid 100g, cornstarch 50g, gelatin 7.5g, Avicel (microcrystalline cellulose) 25g, magnesium stearate 2.5g. The above (±)-5-[[2-(mercaptomethyl)-
1-Oxo-3-phenylpropyl]amino]pentanoic acid and cornstarch are mixed with an aqueous gelatin solution. The mixture is dried and ground to form a fine powder and granulated. The granules are mixed with Avicel and then magnesium stearate. This mixture was compressed in a tablet press to formulate 1000 tablets containing 100 mg of active ingredient per tablet. Tablets were prepared in the same manner using the compounds of Examples 1-4 and 6-45 as active ingredients.

Claims (1)

[Claims] 1 formula, [In the formula, R ₁ is hydrogen or [formula] R ₂ is [formula] [formula] [formula] [formula] [formula] [formula] [formula] [formula] [formula] [formula] or [formula] R ₃ is hydrogen , lower alkyl,
[Formula] [Formula] [Formula] [Formula] [Formula] [Formula] [Formula] or -(CH ₂ ) _p -cycloalkyl, R ₄ is hydroxy, lower alkoxy,
[Formula] [Formula] [Formula] [Formula] [Formula] [Formula] -O-(CH ₂ ) [ _p+1 ] - OH, [Formula] or [Formula] n is an integer from 1 to 15, R _{5 is} lower alkyl; Carbon number 1
~4 lower alkoxy, lower alkylthio having 1 to 4 carbon atoms, hydroxy, Cl, Br, F, amino, -
NH-lower alkyl (1 to 4 carbon atoms), -N (lower alkyl) ₂ (lower alkyl has 1 to 4 carbon atoms), nitro or trifluoromethyl, r is an integer from 1 to 3 (provided that R ₆ is hydroxy ,
r is 1 only for methyl, methoxy, Cl or F
R ₁₀ is hydrogen, lower alkyl, cycloalkyl or phenyl, R ₁₁ is hydrogen, lower alkyl, lower alkoxy or phenyl, and R ₇ and R ₈ are each independently hydrogen, lower alkyl, [Formula] [Formula] [Formula] [Formula] or [Formula] or R ₇ and R ₈ are both combined with the N-atom to form the formula: (q is 0 or 1, A is CH-R ₉ , oxygen or N
_-R9 and _R9 form a ring (hydrogen or lower alkyl having 1 to 4 carbon atoms), or a pharmaceutically acceptable salt thereof. 2 R ₁ is hydrogen, [Formula _] or [Formula] R ₂ is [Formula] [Formula] [Formula] [Formula] [Formula] [Formula] or [Formula] Chain or branched alkyl, [Formula] [Formula] [Formula] [Formula] [Formula] [Formula] or [Formula] m is 1 or 2, R ₆ is methyl, methoxy, Cl, F, hydroxy,
nitro or amino, _R4 is hydroxy, methoxy, _-NH2 or _R10 is hydrogen, straight or branched lower alkyl having 1 to 4 carbon atoms or cyclohexyl, _R11 is straight or branched lower alkyl having 1 to 4 carbon atoms, and n is an integer of 1 to 9. A compound according to item 1 above. 3. The compound according to item 1 or 2 above, wherein n is an integer of 3 to 6. 4. The compound according to item 1 or 2 above, wherein n is 1. 5. The compound according to item 1 or 2 above, wherein n is 2. 6. The compound according to item 1 or 2 above, wherein n is 3. 7. The compound according to item 1 or 2 above, wherein n is 4. 8 The compound according to the first term or the second term, wherein n is 5. 9 The compound according to the first term or the second term, wherein n is 6. 10 The compound according to item 1 or 2 above, wherein n is 9. 11. The compound according to any one of Items 1 to 10 above, wherein R ₁ is hydrogen. 12. The compound according to any one of Items 1 to 10 above, wherein R ₁ is [Formula]. 13 The above-mentioned items 1 to 1 in which R ₂ is benzyl
A compound according to any one of Item 2. 14. The compound according to any one of items 1 to 13 above, wherein R ₃ is hydrogen. 15 The above-mentioned items 1 to 1, wherein R ₃ is benzyl
A compound according to any one of Item 3. 16. A compound according to any one of clauses 1 to 15 above, wherein 16 R ₄ is hydroxy. 17 The above-mentioned items 1 to 1 in which R ₄ is methoxy
A compound according to any one of Item 5. 18 Items 1 to 15 above, wherein R ₄ is -NH ₂
A compound according to any one of paragraphs. 19 formula, [In the formula, R ₁ is hydrogen or [formula] R ₂ is [formula] [formula] [formula] [formula] [formula] [formula] [formula] [formula] [formula] or [formula] R ₃ is hydrogen , lower alkyl,
[Formula] [Formula] [Formula] [Formula] [Formula] [Formula] [Formula] or -(CH ₂ ) _p -cycloalkyl, R ₄ is hydroxy, lower alkoxy,
[Formula] [Formula] [Formula] [Formula] [Formula] [Formula] -O-(CH ₂ ) [ _p+1 ] - OH, [Formula] or [Formula] n is an integer from 1 to 15, R _{5 is} lower alkyl; Carbon number 1
~4 lower alkoxy, lower alkylthio having 1 to 4 carbon atoms, hydroxy, Cl, Br, F, amino, -
NH-lower alkyl (1 to 4 carbon atoms), -N (lower alkyl) ₂ (lower alkyl has 1 to 4 carbon atoms), nitro or trifluoromethyl, r is an integer of 1 to 3 (provided that R ₆ is hydroxy ,
methylyl, methoxy, Cl or F), R ₁₀ is hydrogen, lower alkyl, cycloalkyl or phenyl, R ₁₁ is hydrogen, lower alkyl, lower alkoxy or phenyl, and R ₇ and R ₈ are Each independently hydrogen, lower alkyl, [Formula] [Formula] [Formula] [Formula] or [Formula] or R ₇ and R ₈ together with the N-atom form the formula: (q is 0 or 1, A is CH-R ₉ , oxygen or N
-R ₉ and R ₉ form a ring of hydrogen or lower alkyl having 1 to 4 carbon atoms] or a pharmaceutically acceptable salt thereof as an active ingredient. agent.