JPS6140252A - N-carbamoylamino acid derivative - Google Patents

Abstract

NEW MATERIAL:N,N-[N-Methyl-N-(2-phenethyl-2-carboxyethyl)carbamoyl]-benzyl alycine expressed by formula I, a salt or ester thereof. USE:A hypotensor and/or a remedy for heat failure, having powerful inhibitory action on angiotensin converting enzymes. PREPARATION:According to reaction formulae, a beta-amino-acid derivative expressed by formula II is reacted with phosgene to produce an N-chlorocarbonylamino acid derivative, and the resultant product is reacted with an alpha-amino acid derivative expressed by formula III to obtain an N-carbamoylamino acid derivative. The resultant product is further hydrolized in a solvent such as hydrous methanol using an alkali such as NaOH or KOH to obtain the compound expressed by formula I. In said reaction, the phosgene is used in an equimolar amount or more, usually 1.5-15 times the molar amount of the compound expressed by formula II.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to an N-carbamoyl amino acid derivative represented by general formula (I) or a pharmacologically acceptable salt or ester thereof, and medical uses thereof.

The compound of the present invention represented by the above general formula [1] is a novel compound, has a strong inhibitory effect on angiotensin converting enzyme, and is a useful pharmaceutical compound that can be used for the treatment of hypertension, heart failure, etc.

Angiotensin converter # is an enzyme that acts on angiotensin-I in the body and converts angiotensin-■ into angiotensin-H, which is a substance that increases blood pressure. Furthermore, it is an enzyme that acts on the in vivo antihypertensive substance pradikinin and inactivates it. Angiotensin converting enzyme does both of the above and increases blood pressure. Therefore, inhibiting angiotensin-converting enzyme activity causes a decrease in blood pressure and is extremely useful in treating hypertensive patients and ischemic heart failure patients.

Conventionally, compounds that have an inhibitory effect on angiotensin converting enzyme include peptides found in snake venom and 2-
Methyl-3-thiopronoyl-L-proline is known. The former does not exhibit a phlegm-inhibiting effect upon oral administration. In addition, the latter has side effects such as rash, taste disturbance, and kidney damage.As a result of detailed research into the relationship between the molecular structure and inhibitory activity of compounds that have an inhibitory effect on angiotensin converting enzyme, the present inventors found that the general formula We discovered that a new compound represented by CI) has excellent inhibitory activity,
This has led to the present invention.

The pharmaceutically acceptable salts of the present invention are suitable inorganic or organic salts of at least one carboxylic acid that are non-toxic. Bases that can be used to produce the pharmacologically acceptable salts include hydroxides, carbonates, and bicarbonates of alkali metals or alkaline earth metals such as sodium, potassium, calcium, and magnesium; or lysine. , ornithine, dicyclohexylamine, and other organic bases.

The pharmacologically acceptable esters of the present invention are lower alkyl esters of at least one carboxylic acid that are non-toxic. The lower alkyl group is a straight or branched alkyl group having 1 to 4 carbon atoms, such as methyl and ethyl.

Examples include n- or isopropyl and n- or isobutyl groups.

Compound (1) of the present invention has one asymmetric carbon atom in the molecule and has 2wi optical isomers, and the present invention encompasses both of them.

Compound (1) of the present invention can be synthesized by the method shown by the following reaction formula.

First, a β-amino acid derivative [formerly] is reacted with phosgene to form an N-chlorocarbonyl amino acid derivative in step-a.
), and by reacting this with α-amino acid conductor 1], an N-carbamoyl amino acid derivative OW (process a-b), which is the ester conductor of the compound of the present invention, can be obtained.

By hydrolyzing this N-carbamoyl amino acid derivative [■] (Step-○), the compound of the present invention represented by the formula m is obtained.

The step-a is carried out by dropping an organic solvent solution in which the compound [■] and the deoxidizing agent are dissolved into an organic solvent solution of phosgene at a temperature of 0° C. or lower, preferably -20° C. or lower and stirring. It is possible. After the reaction is completed, unreacted phosgene is removed by vacuum distillation or the like to obtain N-chlorocarbonyl amino acid derivative 4. Step-a is diluted with an organic solvent in the same manner as step-a, and an organic solution of compound (III) and deoxidizing agent is added to the solution of the 7'CN-chlorocarbonyl amino acid derivative at a temperature of 20 to 40°C. This is done by dripping and stirring. The reaction is usually completed in 5 to 20 hours.

The organic solvent is not particularly limited as long as it does not participate in the reaction, but methylene chloride, tetraroform, tetrahydrofuran, dioxane, etc. can usually be used. Examples of the acid scavenging agent include organic tertiary bases such as triethylamine, N-methylmorpholine, and pyridine. Phosgene is more than equimolar to β-amino acid derivative [■],
It is usually used 1.5 to 15 times. The desilting acid agent is used in Step-1
In step-b, 1 to 2 times the mole relative to the β-amino acid derivative [■], and in Step-b, 1 to 2 times the mole relative to the α-amino acid derivative C1) (if compound (Ill) is a hydrochloride,
Furthermore, equimolar amounts of a de-acidifying agent are used for neutralization.

The organic solvent may be used in an amount sufficient to uniformly stir the reaction system, but it is usually used in an amount of 5 to 100 times the amount of the amino acid derivative.

In addition, in step-a and step-b, compound (II)
and compound [111) may be replaced. Step-C is a hydrolysis reaction of compound [■]. The hydrolysis reaction is usually carried out at room temperature using an alkali such as sodium hydroxide or potassium hydroxide in a water-containing solvent such as water-containing methanol or water-containing ethanol. After completion of hydrolysis, extraction, recrystallization, hydrochromatography, etc. 10 Usually 0 Isolation and purification operations Kx'' compound [1) e?1m is obtained.

N-methyl-β-amino acid ester [n], which is a raw material for the compound [■] of the present invention, can be synthesized, for example, as follows.

In the formula, R represents a phenethyl group.

It should be noted that the above synthesis method shows only one embodiment for obtaining the compound of the present invention, and therefore, the method for producing the compound of the present invention should not be limited thereto.

The compound of the present invention [■] and its pharmacologically acceptable salts and esters were administered to JCL-ICB mice at 300%
When administered orally at a dose of 0ff1g/Kf, it does not show any abnormalities and has a strong inhibitory effect on angiotensin converting enzyme, so it is a useful pharmaceutical compound as a hypotensive agent and/or a therapeutic agent for heart failure. is.

Compound [1) of the present invention can be administered orally, enterally, or by injection in various formulations as a composition together with a pharmaceutically acceptable carrier and/or adjuvant.

At this time, the compounds of the present invention may be used in combination of two or more kinds,
It may also be used in combination with narcissus pharmaceutically active substances.

The formulation of the above composition of the compound of the present invention includes tablets,
It can take various forms such as sublingual tablets, powders, capsules, troches, aqueous or oily solvents, Fa suspensions, emulsions, syrups, and aqueous or oily injections.

In either case, auxiliary agents such as vehicles, carriers, excipients, binders, preservatives, stabilizers, and flavoring agents that are commonly used in pharmaceutical preparations can be added. These adjuvants include, for example, starch, lactose O carrier, excipients such as calcium diphosphate; binders such as tragacanthum, gum arabic, cornstarch, gelatin; disintegrants such as alginic acid; lubricants such as nistearic acid. : Sweeteners such as sucrose; Flavoring agents such as mint, etc. can be used.

The content of the compound of the present invention in the pharmaceutical composition may vary as appropriate, but is preferably 0.01% to 100%, preferably 0.05%.
~80 rice cakes.

The bioactive agent of the present invention can be administered orally or parenterally to humans and animals, but oral administration is preferred.

Oral administration includes sublingual administration. Parenteral administration includes injection administration (e.g. subcutaneous, intramuscular or intravenous injection).

(intravenous drip), rectal administration, etc.

Administration of the bioactive agent of the present invention When the human subject is generally administered orally, the body weight is 1 kiloliter, and the daily commerce is 0.
1 to 500 kg, preferably 0.5 to 20019. For parenteral administration, 0.01 to 200 yai, preferably 0.1 to 100 Mf, is administered in 1 to 4 divided doses, but it may be administered to animals or humans, depending on age, age, individual differences, medical conditions, etc. In some cases, doses outside the above-mentioned range may be administered.The properties, production methods, and pharmacological actions of the compounds of the present invention will be specifically explained below by way of examples.

Example Phosgene 25F TetrahyPloalane (THF) 100
Compound formula (1) at -78C in a solution dissolved in d
4.7 F (2,00 X 10-" mol) and 1 sm (1,08 X 10-" mol) of triethylamine in TH
A solution containing dissolved F 50 dK was added dropwise over 30 minutes under a nitrogen atmosphere, and the mixture was further stirred for 1 hour. After 1 hour, excess phosgene and THF were distilled off under reduced pressure, and THF Zo was added to the residue.

Added d. (R suspension) This suspension was stirred under water cooling, and N~benzylglycine ethyl ester compound formula (1) 4. OJ'(2,0
7X10 mol) and triethylamine 3.2 II
A solution of Ig (2,29×10 mol) dissolved in THF 100− was added dropwise over 30 minutes under a nitrogen atmosphere, and the mixture was stirred at room temperature for 16 hours.

After evaporating THF under reduced pressure, 200 d of ethyl acetate was added to the residue, washed successively with saturated NaHCO@, 10% HCL, H, and 0, and dried over magnesium.

After drying, ethyl acetate was distilled off under reduced pressure, and the residue was purified by medium pressure column chromatography (ethyl acetate:hexane-1:3) to obtain 4.64t of the compound of formula (IV) as a colorless oil.

34N205 calculation value for C26: C: 6B, 70.
H: 7.54, N: 6.16 Measured value: C: 68.
07, H near, 61%N: 6.00 Molecular weight: 45
4.57 Nuclear magnetic resonance spectrum of the obtained compound [■] and
The infrared absorption spectra are shown in FIG. 1 and FIG. 2, respectively.

Compound formula (IV)2. Of (4.40 mmol) was dissolved in 20 m of ethanol, and 1 ml of sodium hydroxide was added therein.
, Of (2,5×10 −2 mol) dissolved in 20 lj of water was added and stirred at room temperature for 2 hours.

After evaporating ethanol under reduced pressure, 50% of water was added to the residue, and the mixture was washed with ethyl acetate. The aqueous layer was adjusted to pH 1 with concentrated hydrochloric acid, extracted with ethyl acetate, washed with saturated NaCJ, and dried over i-gnesium sulfate.

After drying, ethyl acetate was distilled off under reduced pressure, n-hexane was added and crystallized, and the compound formula (I) of the present invention was obtained as white crystals (rn
p44-47°) and 1.50 f was obtained.

C22HmC22H calculated value: C: 66.32, H:
6.5B, N near, 03 measurement value: C: 66.75,
H: 6.7B, N: 7.00 Molecular weight: 398.4
6 Nuclear magnetic resonance spectrum of the obtained compound (I) of the present invention,
and infrared absorption spectra are shown in Figures 3 and 4, respectively.Experimental example Angiotensin converting enzyme inhibitory effect (in vitro
): Rabbit lung extract was extracted and purified and the inhibitory activity was measured using nine angiotensin converting enzymes (Kokufu + Bioch
em.

Blophy8. Acta,, 452. p 14
4-150 (1976)). 14C-labeled hipryl histidyl leucine (specific radioactivity 40 μCV m M ) as a substrate was dissolved in 0.1 M 1j:y acid buffer (pH 8,3) containing 0.3 M NaCl, and 450 μj of this solution was dissolved.
Pour into a test tube and add 40% of the sample compound (inhibitor) solution to the tube.
After adding μ, the temperature was set to 370.

Furthermore, 10 μJ of the enzyme solution was added and the enzyme reaction was carried out at 37 C for 20 minutes. The initial concentration of substrate was 5mM.

After 20 minutes, 0.5-IN hydrochloric acid was added to stop the enzymatic reaction, and the C-hydrochloric acid produced by the decomposition of the substrate by angiotensin-converting enzyme was extracted with ethyl acetate and collected in a liquid scintillation counter. The enzyme activity was determined by quantitative determination. Inhibitory activity was determined by the following formula.

Inhibition rate (%) = 100 (1-T/C)C: Amount of hiplic acid produced when no inhibitor is added.

T: Amount of liquid acid produced when inhibitor is added.

ID inhibits angiotensin converting enzyme by 50%.
The ID50 of the compound of the present invention was 0.093×10 M/-, which represents the concentration of inhibitor that causes harmful effects.

Formulation Example 1 N,N-(N-methyl-N-(2-7enethyl-2-carboxyethyl)cal 10 heavy fl pamoyl]-benzylglycine heavy magnesium oxide 15 parts by weight milk
75 parts by weight of sugar was mixed uniformly to obtain powder and fine granules. This powder was then placed in a gelatin capsule container to form a capsule.

Formulation Example 2 N,N-(N-methyl-N-(2-phenethyl-2-carboxyethyl)cal 45 parts by weight Pamoyl]-benzylglycine starch 10 parts by weight Milk
Sugar 20 parts by weight Polyvinyl alcohol 3 parts by weight Water
After uniformly mixing and kneading 22 parts by weight, the mixture was pulverized and granulated, dried and sieved to obtain granules.

Formulation Example 3 N,N-(N-methyl-N-(2-7enether-2-carboxyethyl)cal 0.6 tS/moyl)-
99.4 parts by weight of benzyl chrysine physiological saline was added, mixed under heating, and sterilized to obtain an injection.

[Brief explanation of drawings]

FIG. 1 and FIG. 2 respectively show the nuclear magnetic resonance spectrum and infrared absorption spectrum of the ester derivative side of the compound of the present invention obtained in the Examples. FIG. 3 and FIG. 4 respectively show the nuclear magnetic resonance spectrum and the infrared absorption spectrum of the compound [1] of the present invention obtained in the Examples.

Claims (2)

[Claims] (1) An N-carbamoyl amino acid derivative represented by the general formula ▲ Numerical formula, chemical formula, table, etc. ▼ or a pharmacologically acceptable salt thereof or ester thereof. (2) At least one of the N-carbamoyl amino acid derivatives, pharmacologically acceptable salts thereof, or esters thereof represented by the general formula ▲ Numerical formulas, chemical formulas, tables, etc. ▼
Angiotensin converting enzyme activity inhibitor containing seeds as the active ingredient.