JPH0518813B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to the following general formula (I): The present invention relates to a novel absorption-imparting agent or absorption-enhancing agent containing at least one of the following phenylalanine derivatives or non-toxic salts thereof as an active ingredient. However, in the formula, R ¹ is a hydrogen atom, a halogen atom such as a fluorine atom, a nitro group, a hydroxyl group, or an esterified-protected hydroxyl group, such as a benzyloxycarbonyloxy group, X is CO or SO ₂ , and -Y- is directly a bond, a lower alkylene group, a substituted or unsubstituted vinylene group, such as a fluorine-substituted vinylene group, or a formula -CH ₂
A group represented by -O- or -O-CH ₂ -,
R ² is substituted (e.g. halogen such as chlorine, nitro group,
substituted with a lower alkyl group such as methyl, ethyl, trifluoromethyl, or a lower alkyloxy group such as methoxy) or an unsubstituted phenyl or naphthyl group, or in formula (I), R ² -Y-CO is N-
Benzyloxycarbonylphenylalanyl, N
-benzyloxycarbonyl-4-halogeno(e.g. fluoro)phenylalanyl, or N-(m
-Methoxycinnamoyl)phenylalanyl group, respectively. That is, when the above-mentioned phenylalanine derivative is administered together with, for example, a pharmaceutically active substance, the pharmaceutically active substance is absorbed orally or parenterally (eg, rectally), or its absorbability is increased. The phenylalanine derivative represented by the formula (I) is unique; for example, phenylalanine itself or N-acetylphenylalanine, lower alkyl esters or amides of the acid represented by the formula (I), etc. Neither is useful as an absorbency-imparting agent. In addition, some of the phenylalanine derivatives represented by formula (I) are known and some are new, but they can be obtained by using conventional N-acylation reactions, salt formation processes, and the production examples described below. It can be prepared. The phenylalanine derivative or its salt represented by the following general formula () is a new compound. (However, in the formula, R ¹ , R ² , and Y have the same meanings as above.) Note that the phenylalanine derivatives represented by the above formulas () and (I) have an asymmetric carbon atom. However, depending on the combination of substituents, D-form, L-form,
Any DL type is acceptable. The above phenylalanine derivative may be in the form of a salt. For example, metal salts such as sodium, potassium, lithium, calcium, etc., or organic bases such as ammonia, dicyclohexylamine, N-
Examples include salts with basic amino acids such as methyl-D-glucosamine, lysine, and arginine, and pharmaceutically acceptable salts are used for pharmaceutical purposes. The above-mentioned phenylalanine derivative or a pharmaceutically acceptable salt thereof can be administered orally or parenterally (for example, rectally) together with, for example, a pharmaceutically active substance to impart absorption properties. For example, in the case of insulin, it exhibits an oral or rectal absorption effect and an inhibitory effect on trypsin and/or chymotrypsin, which are proteolytic enzymes that degrade insulin. In particular, in insulin therapy for diabetes, oral or rectal insulin treatment has not reached the level of practical use, and there is a clinical need to develop insulin administration and absorption agents that are highly safe even for long-term use. Therefore, there are extremely high expectations for the present invention. The pharmaceutically active substance may include, for example, two or more hydrophobic amino acid residues that can form a spatial configuration that allows non-covalent interaction with the adjuvant (phenylalanine derivative or its salt). , polypeptides or derivatives thereof that exist in close proximity or in aggregation at one or more locations, and analogs of both groups. Furthermore, specific examples include water-soluble globular proteins that have a collection of hydrophobic amino acid residues on their surface (insulin, insulin-like growth factor I (IGF-I), insulin-like growth factor (IGF-), pancreatic polypeptide etc.); In a cyclic peptide aqueous solution in which two or more hydrophobic amino acid residues exist adjacently or in close proximity, in the dominant conformation formed in the coexistence of the absorption-imparting agent used in the present invention, two or more hydrophobic amino acid residues Examples include those in which the amino acid residues are close to each other or clustered together at one or more locations. The absorption-imparting agent used in the present invention may be used in an amount of 0.1 to 2000 mg, preferably 0.2 to 500 mg, per 25 units of the drug, such as insulin. The absorption-imparting agent and the pharmaceutically active substance may be used, for example, in the form of tablets, capsules, elixirs, microcapsules or suspensions. The phenylalanine derivative of the present invention is administered per patient to patients who require treatment together with the above medicines.
A total daily dose of 0.2 to 2000 mg per day can be administered in a dosage range of 0.1 to 1000 mg, generally divided into several doses. Dosage will vary depending on the severity of the illness, the weight of the patient and other factors recognized by the administerer. Typical combinations set forth above are formulated into pharmaceutical compositions as discussed below. Approximately 0.2 to 500 mg of the compound or mixture of the phenylalanine derivative or physiologically acceptable salt used in the present invention may be present in a physiologically acceptable vehicle, carrier, excipient, binder,
It is incorporated in unit dosage form with preservatives, stabilizers, flavoring agents, etc. as required by generally accepted pharmaceutical practice. The amount of active substance in these compositions or preparations is such that a suitable dosage within the indicated range will be obtained. Specific drugs that can be mixed into tablets, capsules, etc. are shown below. Binders such as tragacanth, gum arabic, corn starch or gelatin; Excipients such as microcrystalline cellulose; Corn starch, pre-gelatinized starch,
leavening agents such as alginic acid; lubricants such as magnesium stearate; sweetening agents such as sucrose, lactose or saccharin; flavoring agents such as peppermint, redwood oil or thierry, when the dosage unit form is a capsule. Materials of the type described above may further contain a liquid carrier such as an oil or fat. Various other materials can be present as coatings and to otherwise modify the physical form of the dosage unit. For example, tablets may be coated with syrup, sugar or both. A syrup or elixir is an active compound,
It may contain sucrose as a sweetening agent, methyl and propylparabens as preservatives, dyes and flavoring agents such as thiery or orange flavor. Particularly for insulin, it is preferable to use enteric-coated preparations. For example, an 8% aqueous solution of hydroxyphenylmethylcellulose is used as a coating pretreatment agent, and a 10% aqueous solution of hydroxypropylmethylcellulose phthalate and a 3% aqueous solution of polyacetin are used as coating agents, respectively. Enteric-coated preparations may be prepared using conventional methods. Specific examples of phenylalanine used in the present invention are as follows.

[Table] Lanin

[Table] Nylalanine

[Table] Production examples are shown below. Production example 1 (S-6, ) 2.1 g of 4-nitro-L-phenylalanine was added to 10
% NaOH 10ml and stirred at room temperature with 1.7g ether solution of phenoxyacetyl chloride,
A solution of 2.7 g of Na ₂ CO ₃ in 25 ml of water was added dropwise alternately over 20 minutes. After stirring for 3 hours at room temperature, the mixture was acidified with dilute hydrochloric acid to precipitate crystals. Filter this out,
After washing with water, the crystals were recrystallized from dioxane to obtain 2.4 g of needle crystals of N-phenoxyacetyl-4-nitro-L-phenylalanine. mp147℃ Elemental analysis value (%) C H N Calculated value 59.30 4.68 8.14 Actual value 59.47 4.51 8.03 Production example 2 (S-8, 25 g of 4-phenylbutyric acid was dissolved in 500 ml of chloroform, and 17.3 g of N-hydroxysuccinimide was added. Next, 31 g of N,N'-dicyclohexylcarbodiimide was gradually added while stirring on ice, and after stirring for 1 hour, the mixture was further stirred at room temperature for 7 hours.
After adding 10 ml of glacial acetic acid and stirring for 1 hour, insoluble matter was filtered off, and the filtrate was dried under reduced pressure. The residue was recrystallized from ethyl acetate to give 4-phenylbutyric acid N at mp 82°C.
-35 g of hydroxysuccinimide ester were obtained. A solution of 13 g of the above ester dissolved in 200 ml of chloroform was mixed with 16.5 g of L-phenylalanine and Na ₂
15.9 g of CO ₃ was dissolved in 150 ml of water, and the solution was stirred and dropped at room temperature. After further stirring for 7 hours, insoluble materials were removed by filtration, the aqueous layer was separated, the pH was adjusted to 1.0 with 6N hydrochloric acid, and the precipitated crystals were collected by filtration, washed with water, and recrystallized from 90% methanol water to obtain N- 11.2 g of 4-phenylbutyroyl-L-phenylalanine was obtained. mp178°C Elemental analysis value (%) C H N Calculated value 73.28 6.79 4.49 Actual value 73.24 6.94 4.46 [α] ²⁶ _D +8.33° (C=1, acetone) The following compound was produced by the same method as above.

[Table] Production Example 3 (Production of S-11 and 12) L-phenylalanine (0.1 mol) was added to 2N-
Dissolve in NaOH (50ml) and add 20ml of ether.
While stirring vigorously under ice-cooling, add the corresponding naphthoxyacetic acid chloride (0.1 mol) and 2N-NaOH (100 mol).
ml) was added in portions, stirred at room temperature for 3 hours, washed once with ether, and the aqueous layer was diluted with 4N-HCl.
When the pH is adjusted to 2, coarse crystals will precipitate. This is collected by filtration, dried, and then recrystallized from acetic ether-petroleum ether. The results shown below were obtained. Compound "S-12" was produced by the same method as above.

[Table] Production Example 4 (Production of S-21, 22, 23 and 32) 17.3 g of D-phenylalanine and 17.6 g of sodium hydrogen carbonate were added to 150 ml of water, and to this mixture was added N-benzyloxycarbonyl with stirring. A solution of 27.7 g of -L-phenylalanine N-hydroxysuccinimide ester dissolved in 150 ml of tetrahydrofuran was added at room temperature and reacted overnight. 200 ml of water was added to the reaction solution, and the pH value was adjusted to 2 with cooled 4N hydrochloric acid. After extraction with 500 ml of ethyl acetate, the resulting organic layer was washed with 1N hydrochloric acid, then with saturated saline, and then dried over anhydrous sodium sulfate. The residue (28.0g) obtained after distillation under reduced pressure was dissolved in ethyl acetate.
The target product (S-
21) was obtained in an amount of 20 g (yield: 65%). Other compounds were similarly produced.

[Table] Note that the phenylalanine derivatives used in the present invention are non-toxic or have low toxicity. The results of oral acute toxicity measurements for some of the compounds are shown. 0.5% sample
Female ICR− suspended in CMC sodium salt solution
It was orally administered to CD1 mice and observed 24 hours later. Compound (mgKg) -3 〉2500 S-5 〉4000 S-7 〉3000 S-10 〉3000 S-11 〉4000 S-12 〉4000 S-13 〉3500 S-15 〉2750 S-17 〉750 S- 18 〉4000 The present invention will be specifically explained below using Examples and Test Examples, but the present invention is not limited thereto. Examples and Test Examples The activator according to the present invention shown in Table 2 below was added at 0.5%.
CMC-suspended in 0.05M Tris HC1 buffer (PH7.8), mixed with insulin, and orally administered in a predetermined amount to female ICR-CD1 mice (5 to 7 weeks old), and after a predetermined time, to a control group. The blood glucose lowering rate (%) and the fold increase in blood insulin were measured. The results are shown in Table 2 below. In addition, in Table 2, "Z" in the structural formula means a benzyloxycarbonyl group. In addition, among the effect figures, the upper row shows 1) blood sugar lowering rate (%), and the lower row in parentheses shows 2) insulin increase fold.

【table】

【table】

【table】

[Table] Although the above table describes an example of oral administration, the activator according to the present invention exhibits a similar activating effect when used in combination with insulin in the form of a normal suppository. As explained in detail above, according to the present invention,
The practical application of oral or parenteral (for example, rectal) administration of insulin has been opened, and its effects are remarkable. Tablet Example 0.577 g (15000 units, zinc content 0.5%) of porcine insulin was dissolved in 30 ml of 0.05N hydrochloric acid, and then diluted to 300 ml with distilled water. After dissolving 6 g of compound S-27 in 200 ml of 0.1N sodium hydroxide, the pH value was adjusted to 7.5 with 0.1N hydrochloric acid. Add phosphate buffer (0.02M, PH7.5) to this
was added to dilute to 600ml. While maintaining the S-27 solution at 20° C. and stirring vigorously, the insulin solution was added dropwise to the solution, and after adjusting the pH value to 7.5, it was immediately subjected to freeze-drying. The tablets contained 25 mg of the freeze-dried powder, 82 mg of pregelatinized starch, 82 mg of microcrystalline cellulose, and 1 mg of magnesium stearate.
Enteric-coated preparations are produced by a conventional method using a hydroxyphenyl methylcellulose (8%) aqueous solution as a coating pretreatment agent, and then a hydroxypropyl methylcellulose phthalate (10%) aqueous solution and a polyacetin (3%) aqueous solution as coating agents. did. In a similar manner, other phenylalanine derivatives as described above can be similarly formulated in place of the above phenylalanine derivative (S-27) together with pre-gelatinized starch, microcrystalline cellulose and magnesium stearate. Ta. Capsule Example 30 g of the compound S-22 was added to 250 ml of glacial acetic acid, dissolved by heating, and then cooled to 20°C. While stirring, 2 g of porcine insulin (52,200 units, zinc content 0.5%) was added and dissolved in small amounts. Acetic acid was distilled off under reduced pressure at the same temperature. 100 ml of n-hexane was added to the obtained solid residue, and after pulverization, it was collected by filtration and washed, and the adhering n-hexane was distilled off under reduced pressure. Further, it was subjected to vacuum drying in the presence of solid sodium hydroxide. Dry-filled capsules containing 50 mg of active ingredient per capsule were prepared. The above dry powder 50mg Lactose 149mg Magnesium stearate 1mg Capsule (size No. 1) 200mg The dry powder (50mg) was made into No. 60 powder, passed through lactose and magnesium stearate No. 60 sieve cloth, and passed over the above powder. , then mixed thoroughly. This was filled into No. 1 dry gelatin capsules. A part or all of the powder mixed at this time can also be filled as an enteric preparation as described above. The above results show that the phenylalanine derivatives used in the present invention are useful as agents for imparting or enhancing absorption of, for example, pharmaceutically active substances, particularly insulin.

Claims (1)

[Claims] 1 The following general formula (I): An absorption-imparting agent containing at least one of the following phenylalanine derivatives or non-toxic salts thereof as an active ingredient. However, in the formula, R ¹ is a hydrogen atom, a halogen atom,
Nitro group, hydroxyl group, or esterified protected hydroxyl group, X is CO or SO ₂ , -Y- is a direct bond,
A lower alkylene group, a substituted or unsubstituted vinylene group, or a group represented by the formula _-CH2 -O- or -O- _CH2- , ^R2 is a substituted or unsubstituted phenyl or naphthyl group, or a group represented by the formula ( I) medium, R ² −
Y-CO is N-benzyloxycarbonylphenylalanyl, N-benzyloxycarbonyl-4
-Represents a halogen phenylalanyl or N-(m-methoxycinnamoyl)phenylalanyl group, respectively.