WO1993018013A1 - N,N'-ETHYLENE-BRIDGED DIPEPTIDE COMPOSED OF DIFFERENT OPTICALLY ACTIVE α-AMINO ACIDS AND PRODUCTION THEREOF - Google Patents

Abstract

An optionally protected N,N'-ethylene-bridged dipeptide represented by general formula (I), a salt thereof, and a process for producing the same, wherein R3 represents -H, -CH3, -CH2CH3, -CH2CH2CH3 or -CH2CH2CH2CH3; symbols * and *' represent each an asymmetric carbon atom; and R1 and R2 are different from each other and represent each -CH(CH3)CH2CH3, -CH2OH, (a), (b), (c), -CH3, -CH(CH3)2, -CH2CH(CH3)2, -CH(CH3)OH, -CH2CH2SCH3, -CH2CONH2, -CH2CH2CONH2, -(CH2)4NH2, -CH2COOH, -CH2CH2COOH, -CH2SH, (d) or (e).

Description

 Specification

 N, N'-ethylene-bridged dipeptides composed of different optically active a-amino acids and their synthetic methods

 The present invention relates to a dipeptide derivative having a novel piperazinone ring structure

 Background technology

 Derivatives in which two nitrogen atoms of a di-amino acid condensed dipeptide are linked via an ethylene bridged chain are expected to have their own physiological activity due to their unique structure At the same time, it has attracted attention because it can be used as a structural unit of analogues of bioactive peptides, which are resistant to hydrolysis by enzymes and have immobilized conformation. I was

For example, Japanese Patent Application Laid-Open No. 62-196600 discloses a piperazinone derivative of homodipeptide, but does not disclose a piperazinone derivative in which two amino acids are different. It is very unlikely that adjacent amino acids in a protein are the same, and enzymes that specifically cleave between homodipeptides even at cleavage sites such as peptidases Since it is hardly known, the piperazinone derivative disclosed in this publication is difficult to use in the production of peptide analogs. Also, Hlloon et al., Analysis of in vivo Stability, 6 41-644. (1982), and ϋ. S. Patent No. 4. 251, 438 describe derivatives such as Phe-Leu having a piperazinone skeleton. However, these documents introduce a substituent corresponding to the side chain of amino acid after forming the piperazinone skeleton, so that a mixture of four optical isomers is formed, Among them, only the Ellis mouth and threo are disclosed. Enzymes specifically recognize the configuration of the substrate and the configuration is very important in expressing the activity. Therefore, in such a racemic mixture, the activity and stability of the enzyme in vivo, It is not enough to make modifications such as side effects.

 In addition, J. Chem. Soc. PERKIN TRANS. I, 1687-1689 (1989) describes piperazinone derivatives of dipeptides containing R- and S-Tyr and G1y. However, there is no description of a dipeptide containing two kinds of optically active amino acids and a method of synthesizing the same. Since very few enzymes specifically cleave peptide bonds involving Gy, this document is also poorly applicable to the synthesis of various protein derivatives.

The present invention relates to a novel and optically active dipeptide having a piperazinone ring structure, which is expected to have its own physiological activity and is useful as a constituent unit of a bioactive peptide analog Intended to produce derivatives

 Opening up the invention

 The present inventors have conducted intensive studies to achieve the above object, and as a result, linked the amino groups of two different optically active monoamino acids via an ethylene cross-linked chain, and then used an acid catalyst. It has been found that a desired dipeptide derivative having a novel piperazinone ring structure can be produced by cyclization.

 The present invention provides N, N'-ethylene-crosslinked dipeptides and salts thereof which may have the following protecting groups.

 1. General formula (I)

(Wherein, R ₃ represents a hydrogen atom or an alkyl group having 1 to 6 carbon atoms, * represents two asymmetric carbon atoms of this substance, and R _t and R ₂ represent the following formula (A): CH _3, one _{CH (CH 3) 2, -} . CH 2 CH (CH3) 2, CH (CH,) CH 2 CH 3, one _{CH 2 OH, (CH 2)} 3 NH-C-NH 2 -CH 2 -HQ), CH ₂ H> -OH,

 NH

CH (CH ₃ ) HI H,-CH ₂ CH ₂ SCH ₃ , -CH ₂ CONH ₂ , CH ₂ CH ₂ CONH ₂ ,-(CH ₂ ) ₄ NH ₂ ,-CH ₂ C OH, CH ₂ CH ₂ COOH ,-CH ₂ SH _F -CHJ-J— IvT,

And the substituents represented by the general formula (A), each having a different type of substituent or protecting group. )

N, Ν'-ethylene crosslinked dipeptides and salts thereof which may have a protecting group represented by

2. — General formula (la):

(Wherein R ₃ , * and * ′ are the same as above. ^R _la and ^R 2a are represented by the following formula (B): one (CH), NH-C-NHTo s,

 NH

-CH ₂ OBz l,-(CH ₂ ), NH ~ C-NHN0 ₂ .

NH

 0

-(CH ₂ ) ₄ NHZ,-(CH ₂ ) ₄ NHBo c, one CH ₂ COORb.-CH CH ₂ COORb, one CH ₉ SB z 1, -CHn-i ~ N

 N

B z 1 [Wherein, R and R _k represent a methyl group, an ethyl group,

& D

Groups which may have a substituent such as an alkyl group having 1 to 4 carbon atoms such as a butyl group, a benzyl group, a p-methoxybenzyl group, a p-nitrobenzyl group, and a -chlorobenzyl group. Represents an benzyl group. B z 1 represents a benzyl group which may have a substituent such as a benzyl group, a p-methoxybenzyl group, a p-2-trobenzyl group, and a p-chlorobenzyl group. ] Represents substituents having different types of protecting groups. )

And N, N, and ethylene-crosslinked dipeptides having the protective group according to 1. and salts thereof. 3. —General formula (Ib):

(Wherein R ₃ is the same as above. * And * 'represent the two asymmetric carbons of this material; R _lb and R _2b are represented by the following formula (A): CH3, one CH (CH ₃ ) ₂ , one CH ₂ CH (CHj) ₂

CH (CH ₃ ) CH ₂ CH, .-CH ₂ OH. (CH ₂ ), NH-C-NH ₂ ,

H

CH (CH ₃ ) OH, -CH ₂ CH ₂ SCH _3. One CH ゥ CONH ₂ , CH ₂ CH ₂ CONH,-(CH ₂ ) ₄ NH ^, one CH ₂ COOH,

_{CH 2 CH 2 COOH, - CH} 2 SH, -CH 2 -j- ijl

And each represents a substituent having no different type of protecting group. ) N, N'-ethylene-crosslinked dipeptides and salts thereof having no protective group as described in 1. above.

4.2 kinds of different One optically active a- A Mi Amino Acids NH ₂ one CH (R ₁₎ and one COOH

NH. One CH (R ₂ ) one COOH, and and R ₂ are represented by the following formula (A): . One CH ₃ one CH (CH _{3) 2,} one CH ₂ CH (CH _{3) 2,} one _{CH (CH NH 3) CH 2} CH 3, - CH 2 OH,

NH 1 CH (CH ₃ ) HI H,-CH ₂ CH ₂ SCH ₃ CH 1 CH ₂ CONH ₂ , 1 CH ₂ CH ₂ CONH ₂ , 1 (CH ₂ ) ₄ NH ₂ , 1 CH ₂ COOH,-CH ₂ CH _{2 COOH, - CH 2 SH,} -CH 2 ~ j- NH

And each represents a different type of substituent. Or the α-amino acid having a protecting group in the side chain, and a compound represented by the following general formula (H):

R ₄ CH ₀ CH ₉ R ₅ (H)

CR ₄ and R ₅ are the same or different and represent a leaving group. ) Is reacted in an aqueous solution under alkaline conditions, and then a cyclization reaction is carried out in the presence of an acid catalyst. General formula (I) characterized by performing a deprotection reaction

It is intended to provide a method for producing an N, 1ST-ethylene-crosslinked dipeptide which may have a protective group represented by the formula:

In the present invention, and R ₂ may be each having a protecting group and R _2- , or may be R _lb and R _2h having no protecting group. Is the favored arbitrary and Ϊ _9, the following formula (A '):

-CH,, — CH (CH ₃ ) ₂ , -CH CH (CH,)

3 ^J 2

CH (CH ₃ ) CH ₂ CH ₃ ,-CH ₂ OH,

(CH ₂ ) 3 NH-C-NH ₂ ,

 NH

CH _2- ^) H,-(CH ₂ ) ₄ NH ₂ ,-CH ₂ CO OH,

_{CH 2 CH 2 COOH, - CH} 2 SH, -CH 2 -. HN

-C,-CH (CH ₃ ) OH,-CH ₂ CH ₂ SCH3

-CH ₂ CONH ₂ , -CH ₂ CH ₂ CONH ₂

And substituents which may have different types of protecting groups. More preferably, substituents each having a different kind of protective group represented by the above formula (A ′) are mentioned.o

In the present invention, R ₃ represents a hydrogen atom, a methyl group, Examples thereof include an ethyl group, an n-propyl group, an n-butyl group, an n-pentyl group and an n-hexyl group.

 When the compound of the present invention has a protecting group, the protecting group can be easily deprotected by a method and a method generally used in the field of peptide synthesis.

 In the present invention, monoamino acids that do not form dipeptides include methionine, fenilalanine, tryptophan, lysine, arginine, tyrosine, cystine, aspartic acid, glutamic acid, and the like. 18 types of optically active α-amidin found in common proteins such as asparagine, glutamine, histidine, alanine, rosin, * lin, leucine, isoleucine, serine, and threonine For example, those having a reactive functional group in the side chain may be used in which the functional group is protected.

In the present invention, the α-amino acid having a protecting group or the α-amino acid having R ₂ as a substituent includes α-amino acids having a protecting group on a side chain commonly used in the art. The following are preferably used:

 (1) A guanidino group of Ar g protected by a nitro group or a tosyl group,

(2) The side chain carboxyl group of Glu or Asp is converted to methyl, ethyl, n-propyl, isopropyl, n-butyl, isopropyl An alkyl group having 1 to 4 carbon atoms such as butyl and t-butyl or an optionally substituted benzyl such as benzyl, p-methoxybenzyl, ρ-nitrobenzyl and p-chlorobenzyl Ester,

 (3) Cys thiol group protected with a benzyl group which may have a substituent such as p-methoxybenzyl group, benzyl group or ACM,

 (4) those in which the imidazole group of His is protected by the above-mentioned optionally substituted benzyl group, tosyl group, Fmoc group or Bom group;

 (5) a side chain amino group of Lys protected with a benzyloxycarbonyl group or a t-butoxycarbonyl group; and

(6) Those wherein the hydroxyl group of the side chain of T hr is protected with t tert -butyl group or benzyl group.

 In the present invention, a leaving group represented by R.

 The (leaving group) is not particularly limited, but may be a halogen atom such as a fluorine atom, a chlorine atom, a bromine atom or an iodine atom, or one OT os (Ρ-toluenesulfonic acid ester), one OM s (medium Examples of such groups include ester groups such as tansulfonic acid ester) and -0Ac (acid ester), which can be eliminated by reacting with an amino group.

The salt of the compound of the general formula (I) of the present invention includes p-toluene Inorganic acid salts such as enesulfonic acid, benzenesulfonic acid, hydrochloric acid, sulfuric acid, nitric acid, and perchloric acid; organic acid salts such as trifluoroacetic acid, maleic acid, and fumaric acid; or sodium salts and potassium. And alkali metal salts such as salts. These salts can be produced by a conventional method such as dissolving or suspending the compound of the present invention in a solvent, and adding an acid or a base in an equal or excess amount.

Preferred correct is a combination of amino acids of the compounds of the present invention, for example A RG- E _{(Χ χ, S er, P} he, A 1 a, H is, M et, L eu, I le, G lu, a sp, G ln, a s, C ys, L ys, V al, shows a T yr, T rp or _{T hr), L ys- X 0} (X 2 are, S er, P he, a la , His, Met, Leu, Ile, G1u, Asp, Gln, Asn, Cys, Arg, Val, Tyr, Trp or Thr), _Xg — L eu (Xo is Ser, P he, A la, His, Met, Arg, I le, G lu, A sp, G ln, A sn, Cys, Lys, Va 1, T yr, T rp or T hr), P he—Met;,, T hr—Arg ゝ Ser—A la, A 1a—T rp and the like.

These dipeptide derivatives include, but are not limited to, opioid peptides, bone resorption inhibitory peptides, angiotensin (Angiotensin) I and Π, ΑΝ Ρ, bra Dikinin (Bradykinin), BNP, Canolecithinin (Calcitonin), C GRP, CCK, Conotoxin, CNP, Dynorphin, Elafin (E1 affin), α-Endolphin (fl-Endorphin), 3-endorphin, 7-endorphin (r-En dorphin) endotherin-1 (Endotherin-1), endotherin-1 2 ( Endotherin-2), Endotherin-3 (Endother in-3), Enkephalin (Enkephalin), Demorphin (Dermorp in) Galanin, RGDS, GRGDS, YIGSR, Gastrin ), GRF, GRP, LH-RH, matillin (ttatilin) PTPTH-rp, secretin (Secretin), serum thymic factor (Serum thymic factor), somatostatin (Somatostatin), Substance P (Substance P), TRH Bioactive peptides such as, VIP, Vazopressin De, an analog thereof, are useful for modifying its derivatives, or O Li Gore peptidase de is Furagume down bets from these bioactive Bae flop switch mode. The resulting modified peptides can act on both agonists and antagonists of natural bioactive peptides.

For example, a site that is specifically cleaved by an enzyme such as the above-mentioned physiologically active peptide can be replaced with the peptide derivative of the present invention. 5 makes it less susceptible to degradation by peptidases and is useful for increasing its stability in vivo. In addition, since the dipeptide derivative of the present invention is optically active, it can retain agonist and Z or antagonist activity even when the desired physiologically active peptide or the like is substituted with the derivative. The onset of non-specific toxicity can also be suppressed as compared to the racemic form.

 The dipeptide derivative (I) of the present invention is produced, for example, as follows.

HOOC-CH-ΝΗ ₂ + ΗΛ N-CH-COOH

alkalinity

HOOC-CH-NHCH. CH NH-CH-COOH

(Wherein, R ₂ , R ₃ , R ₄ and R ₅ are the same as above) * First, two different α-amino acids (NH. -CΗR _l ) one COOH and NH ₂ -CH (R) One COOH, (R, and R ₂ are the same as described above))) is represented by the general formula (Π):

R ₄ CH ₂ CH _? R ₅ (Π)

(Wherein, R ₄ and R ₅ are the same as described above) under alkaline conditions.

 As a solvent for the reaction, water, a mixed solvent of water and an alcohol, or a mixed solvent of water and acetonitrile, dioxane, tetrahydrofuran or the like is used, and water is preferably used. Can be The reaction temperature is about 60 to 100, and the reaction time is about 3 to 10 hours. The different amino acids are usually used in equimolar amounts, and the compound of the general formula (π) is used in an amount of about 0.5 mole per 1 mole of either amino acid.

 Under alkaline conditions, aqueous sodium hydroxide, hydroxide hydroxide and other metallic hydroxide aqueous solutions, sodium carbonate, sodium carbonate, hydrogencarbonate It is carried out by adding an alkali metal carbonate such as a stream to the solvent, and the amount added is about 1 to 2 times the mole of 1 mole of the total amino acid used. .

The product of this reaction is a mixture of the following three compounds: Obtained

HOOC— CH— NHCH ゥ CH «NH-CH-COOH,

 I ー 'i I ·

 R,

 (1)

(HOOC-CH-NHCH _2- ) ₂ (2)

(HOOC-CH-NHCH _2- ) ₂ (3)

(Wherein and _RQ are the same as above). The mixture may be separated at this point, but is preferably purified after the next cyclization reaction. The desired compound (I) is obtained by subjecting the above three mixtures or the separated and purified condensate of two different amino acids of (1) to a cyclization reaction in the presence of an acid catalyst in a solvent. . Solvents include methanol, ethanol, and 1-propanol And alcohols such as 1-butanol. Examples of the acid catalyst include p-toluenesulfonic acid, methanesulfonic acid, benzenesulfonic acid, trifluoroacetic acid, concentrated sulfuric acid, hydrochloric acid, and acetic acid. P-toluenesulfonic acid is preferable. The amount of the acid catalyst to be used is about 1 to 3 mol, preferably about 1.5 mol, per 1 mol of the condensate (1) of two kinds of amino acids. The reaction temperature is about 50 to 100, and the reaction time is about 2 to 30 hours.

 When amino acid having a protecting group in the side chain is used as the amino acid raw material, after the cyclization reaction, deprotection is performed according to the conditions of the deprotection reaction generally performed in the relevant life. Perform the reaction. When the main chain carboxylic group is esterified after the cyclization reaction, the ester can be hydrolyzed according to a conventional method.

 The target dipeptide derivative (I) obtained by the above method is purified by a usual purification method, for example, a method such as column chromatography or recrystallization.

 The raw amino acid used in the present invention is an optically active R-amino acid or S-amino acid.

 The dipeptide derivative (I) of the present invention has the following effects.

CD -to o

(1) For example, a combination of feniralanine and methionine The dipeptide is used as a structural unit of obioid peptide.

 (2) Incorporation of this peptide into a site susceptible to hydrolysis by the enzyme can increase the stability of various useful peptides to the enzyme in vivo. In addition, by immobilizing the conformation and increasing lipophilicity due to cyclization, the dipeptide is incorporated into the active site to increase the activity of the physiologically active peptide, or to increase the activity of the peptide having multiple activities. By changing the action pattern and increasing the specific activity, side effects can be reduced.

 (3) The macrocyclic peptide having the above-mentioned dipeptide as a structural unit is useful for inclusion of organic and inorganic compounds because it has a structure similar to cyclodextrin, that is, its cavity has a width. In addition, since it is fat-soluble, it can be used as a reagent for various organic synthesis reactions, and is particularly effective for asymmetric reactions.

Further, according to the production method of the present invention, the cyclization of the difficult and low-yield dipeptide derivative proceeds smoothly, and the dipeptide derivative (I) can be obtained in high yield. In particular, when p-toluenesulfonic acid in alcohol is used as an acid catalyst, the cyclization yield is high, which is preferable. In addition, side reactions are less when α-amino acid having an unstable side chain such as methionine is used as a raw material due to mild reaction conditions. Examples

 Hereinafter, the present invention will be described in more detail with reference to Examples, but the present invention is not limited to these Examples.

Example 1

Dissolve (S)-phenylalanine (0.1mol), (S)-methionine (0.1mol), and caustic soda (0.2m01) in 500ml of water. The aqueous solution is heated to 80. Subsequently, dibromethane (0.1 mol) and potassium carbonate (0.1 mol) are added, and the mixture is further reacted for 5 hours. After the reaction is completed, return to room temperature, and add concentrated hydrochloric acid (0.2 m0 L). The deposited sediment is filtered off. The resulting precipitate, that is, N, N'—a monoamino acid with two ethylene-crosslinked molecules! Of (a) (S)-phenylalanine and (S)-phenylalanine, (b) (S)-methionine and (S)-methionine and (c) (S) one phenylalanine and (S) one methionine] Dry the mixture (about 0.06 mol). The above mixture and P-toluenesulfonic acid monohydrate (0.11 mol) are refluxed for 24 hours in 500 ml of ethanol. After distilling off the ethanol, treat the residue with aqueous sodium hydrogen carbonate solution and extract with dichloromethane. 21 g (approximately 0.05 mO1) of the obtained oily substance was subjected to silica gel chromatography (developing solvent: benzene: ethyl ester ester group). The mixture is separated into four types of N, N'-ethylene-crosslinked dipeptides (1A, 2A, 3A, 4A) by the following method. The production ratio of 1 A, 2 A, 3 A, and 4 A is 1: 3: 2: 1. The structures of 1 A to 4 A are shown below.

General formula

 * R or S and one R or S

1λ; 1ι — CH ₂ I s — CH ₂ CB ₂ SCH 3 (S. S)

2A; Ei — CH ₂ CEz SCE 3. I 2 — CI _2-

4A; ti- ₂ — CE ₂ CI ₂ SCI 3 (S, S)

Example 2

An experiment was conducted in the same manner as in Example 1 in which (R) -phenylalanine and (R) -methionine were combined. The resulting compound has the ID iR ₁ =-CH ₂ ph, R ₂ =-CH ₉

CH ₂ SC Hg (R, R)] and 2 D [=-CH ₀ CH

₂ SCH ₃ , R ₂ = one CH ₂ ph (R, R)]. Here, p li represents a phenyl group.

 1A, 2A, 1D and 2D were purified as the hydrochloride salts (HC1.1A, HC1.2 2, HC1.1D, HC1.2D) by recrystallization from ethanol, respectively. . Table 1 shows the elemental analysis values, melting points, and specific rotations measured in ethanol at room temperature of the above four kinds of dihydroethyl ester hydrochlorides.

 The mass spectrum values are the theoretical values (mZ ζ =

3550).

I— *

 CJin

Table Elemental analysis values of dipeptide ethyl ester hydrochloride clay, »Point and specific rotation Molecular formula Elemental analysis value (%) Melting point Specific rotation Compound (molecular weight) (actual value theoretical value) D

 C H N

H C 1 A 55.86 7.04 7.26 162-5 110

^C 18 ^H 27 ^N 2 ° 3 ^{SC 1}

 (386.9) 55, 87 7.03 7.24

 H C 1 2 A 55, 57 7, 07 7.19

^C 18 ^H 27 ^N 2 ° 3 ^{SC 1} 134-9 99 (386.9) 55.87 7.03 7.24

 H C 1 D 56.15

^C 18 ^H 27 ^N 2 ° 3 ^{SC 1} 7.06 7.22 163-7 + 118 (386.9) 55.87? .03 7.24

 H C 1 2D 55.96

^C 18 ^H 27 ^N 2 ° 3 ^{SC 1} 7.12 7.26 134-9 ', +101 (386.9) 55.87 7.03 7.24

Instead of the combination of (R) -phenylalanine and (R) -methionine, the combination of (R) -phenylalanine and (S) -methionine, or (S) -phenylalanine and ( Similarly, an optically active compound can be obtained by using the combination of R) -methionine.

Example 3

 Using a method similar to Example 1 above,

 (1) (S) —Fenilalanine and (S) —Mouth

 (2) (R) —Fenilalanine and (S) —Mouth Isin

 (3) (S) —Fenilalanine and (R) —Mouth

 (4) An experiment in which (R) -pheniralanine and (R) -mouth was combined was performed in the same manner.

From (1), 1 A 'CR _t =-CH ₂ ph,

R „= one CH ₂ CH CC H ₃ ) ₂ (S, S)) and 2 A 'CR _X =-CH _n CH (CH ₉ ) ₂ , R ₂ =-CH ₂ ph (S, S)],

From (2), IB '[R ₁ = — CH ₂ ph,

R ₂ =-CH ₂ CH (CH ₃ ) ₂ (R, S)] and 2 B '[=-CH ₂ CH (CH ₃ ) ₂ , R ₂ =-CH ₂ ph (S, R)],

From (3), 1 C 'CRj =-CH ₀ ph,

_{2 = - CH 2 .CH (CH} 3) 2 (S, R) ] and 2 CH (CH ₀ ) CH ₂ ph

( ^R 1 = ^-CH 2 3 ^J 2 ^R 2 =

 (R, S)]

From (4), ID '(R ₁ =-CH ₂ ph,

R ₂ =-CH ₀ CH (CH ₃ ) ₂ (R, R)] and 2 D '

CR _t = one CH ₂ CH (C Hg) ₂ , R ₂ = one CH ₂ ph

(R, R)].

 All of these dichlorides are hydrochlorides (H C 1.1A ', H C 1.2 2', H C 1.1D ', H C 1-2D, H C

1 · 1 Β ', HC 1 · 2 Β ⁷ , HC 1 · 1 C, HC 1 ·

Each was purified by recrystallization from ethyl acetate. Table 2 shows the elemental analysis values, melting points, and specific rotations (in ethanol, room temperature) of the eight dipeptidoethyl ester hydrochlorides.

 The mass spectral values are the theoretical values (mZ z-

3 3 2).

C CD

Table 2 Elemental analysis values, melting points, and ratios of dipeptide ethyl ester hydrochloride Molecular formula Elemental analysis value (%) Compound (molecule S) (actual measurement value s¾ value)

 C H N

HC 1 1 A 0 ₇ C 1 61.78 7.95 7, 62

C ₁ ^Η 2 ^Ν 2

 (368.9) 61.86 7.92 7.59

HC 1 2 AC ₁₉ Η ₂₉ Ν ₂₀ , C 1/6 H ₂ 0 61, 30 8, 04 7.55

 (3 7 1.8) 61.38 7, 95 7.53

H C 1 1 D '0, C 1 61.76 7.97 7, 50

C ₁ ^Η 29 ^Ν 2

 (368.9) 61.86 7, 92 7.59

_{HC 1 2 D 'C 19 H} 29 N 2 0 3 C 1/6 H 2 0 61.20 8.01 7. 58

 (37 1.8) 61.38 7.95 7.53

_{HC 1 1 B 'C ig H} 29 N 2 0 3 C 1 61.78 7.90 7.50

 (368.9) 61.86 7.92 7.59

HC 1 2 B 'C ₁₉ H ₂₉ N ₂ 0 ₃ C 1/6 H ₂ 0 61.28 7.90 7.55

 (37 1.8) 61.38 7.95 7, 53

HC 1 1 C 'o ₃ C 1 61.77 7.92 7.48

C 19 ^H 29 ^N 2

 (368.9) 61..86 7.92 7.59

HC 1 2 C 'C _lg H ₂₉ N ₂ 0 _T C 1/6 H ₂ 0 61.41 7.89 7.51

(37 1.8) 61.38 7.95 7.53

Example 4

 Using the same method as in Example 1 described above, various compounds shown in Table 3 below were synthesized.

 Table 3 shows the elemental analysis values, melting points, and specific rotations of the hydrochloride of the compound obtained and the 3,5-dinitrobenzol (DNB) form. The specific rotations in Tables 1 to 3 are values measured at room temperature.

 Table 4 also shows the NMR data of A1a-Trp and Trp-Ala of the present invention.

The NMR data of eAla-Trp and eTrp-Ala in Table 4 are all values measured at 30.0 in CDCIn. Also, * indicates an overlapped signal.

82

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Table 4

NMR data of HC 1 · eA 1 aTr p— OE t

1 ·

Chemical shift (coupling ¾¾) S. ppm (J. Hz) H2 Hll H4i45t H4e & 5e

4.12 1.24 3.26 2.98

 (q, 7.1) (d, 6.7) (ddd, 10.6, 10.6, 4.7) (dt, 11.0,3.4)

 Z75 187

 (ddd, 10.4, 10.1, 3.7) (dt, 1Z 8, 3.7)

H7 H12A & B H14 H15

 5.10 3.45 4.21 1.26

 (dd, 11.0, 5.5) (dd, 15.9. .9) (4, 7.0) (t, 7.0)

 3.34

 (dd, 15.0.11.0)

H① H② H④ H⑤ H⑥ H⑦

 8.78 7.00 7.57 7.09 7.16 7.34

) (S) (d, 7.9) (t.7.0) (t, 7.0) Id.7.) Table 4 (continued) NMR data of r pA I a-OE t

Chemical force shift (number of coupling channels) 5. p pm (J. Hz)

 H2 H11A HUB Η4ι & 5ι H4e 5e

3.78 3.58 3.09 3.27 3.05

 (td, 9.Z 3.7) (dd, 14.7, 3.1) (dd, 147.9.2) (td, 11.0,4.3) (ddd.118, 4.3, 2.5)

 192 3.13 *

(ddd.12.2, 11.0, 4.0) (dt)

H7 H12 HH H15

 5.22 1.36 4.17 1.25

(q, 7.3) (d.7.3) (n) (t, 7.3)

H① H (2> H④ H⑤ H® H⑦

 8.51 7.07 7.69 7.10 7.18 7.35

(S) (d, Z 4) (d, 7.9) U, 7.9) (t, 7.6) (d, 7.6) It has been confirmed that the compounds having a protecting group shown in Table 3 can be deprotected according to a conventional method. In Table 3, "e" in the compound ethyl ester indicates that it is an ethylene-crosslinked dipeptide of the present invention.

 From the results of Table 1, Table 2, Table 3 and Table 4, it was revealed that N, N ′ ethylene-bridged dipeptide derivatives can be obtained in high yield by using the method of the present invention.

Claims

Range of request

 ① General formula (I)

(Wherein, R ₃ represents a hydrogen atom or an alkyl group having 1 to 6 carbon atoms, * and * 'represent two asymmetric carbon atoms of this substance, and R t and R ₂ are represented by the following formula (A): NH 1 CH _3. -CH (CH ₃ ) ₂ , 1 CH ₂ CH (CH ₃ ) ₂ ,-CH (CH,) CH, CH,,-CH ゥ OH, ²

 H

-(CH ₂ ) 3 NH-C-NH ₂ ,-CH ₂ → Q>, CH _2- (Q) H.

 NH

CH (CH ₃₎ shed H, one CH ₂ CH ₂ SCH _3, one CH ₂ CONH _2, CH © CH ₂ CONH. One _{(CH 2) 4 ΝΗ ¾.} -CH 2 COOH.

_{- CH 2 CH 2 COOH, -} CH 2 SH, -CH 2 - | shows a j- each represented by IJ _f different kinds of substituents. ) N, N'-ethylene-crosslinked dipeptides and salts thereof which may have a protecting group represented by ② General formula (la)

(In the formula, R ₃ represents a hydrogen atom or an alkyl group having 1 to 6 carbon atoms, * and * 'represent two asymmetric carbons of the-substance, and R and R _2a are represented by the following formula (A ):

-CH,, -CH (CH ₃ ) ₂ , one CH ₂ CH (CH ₃ ) ₂ , -CH (CH,) CH ₂ CH ₃ , -CH ₂ OH, one (CH ₂ ) 3 NH-C-NH ₂ . — CH ₂ ~. CH ₂ ^ H) H,

NH one CH (CH ₃ ) OH, — CH ₂ CH SCH _3. One CH? CONH ₂ ,

-CH CH ₂ CONH ₂ ,-(CH ₂ ) ₄ H ₂ , one CH ₂ COOH,

-CH ₂ CH ₂ COOH, -CH ₂ SH, -CH _2- || —

 N

H

And each represents a different type of substituent. )

The N, N ethylene-bridged dipeptide and a salt thereof having no protecting group according to claim 1 represented by the formula:

③ General formula (lb)

(Wherein, R ₃ represents a hydrogen atom or an alkyl group having 1 to 6 carbon atoms, * and * 'represent two asymmetric carbons of this substance, and R and R _9b are represented by the following formula (B):

I (CH ゥ) NH— C I NHTo s,

^{L i} II

 NH

-CH ₂ OBz 1,-(CH ₂ ) ₃ NH-C-NHN0 ₂

NH

One (CH ₂ ) ₄ NHZ,-(CH ₂ ) ₄ NHBo c, One CH ₂ COORb, -CHo CH ₂ COORb.

-CH ₂ SB z 1. -CH _2- ^ —

 N

I

 B z 1

And the substituents having different types of protecting groups, respectively. )

2. The N, polyethylene glycol-crosslinked dipeptide having a protecting group according to claim 1 represented by the formula: and a salt thereof.

④ and _RQ are given by the following formula (A-1) -CH,, -CH (CH ₃ ) 2, -CH CH (CH)

3 ^J 2

CH (CH ₃ ) CH ₂ CH ₃ , -CH, OH,

 H N

(CH), NH-C-NH «,

L ⁵ II ¹

 NH

^CH 2 "~ ° ^H 'One ^(CH 2) 4 ^{NH 2'- CH} 2 ^C00H '

_{CH 2 CH 2 COOH, - CH} 2 SH, -CH 2 -j- 1 ^,

One CH 2 "j- ~, -CH (CH ₃ ) OH,-CH NH ₂ CH ₂ SCHj

One CH ₂ CONH ₂ , -CH CH ₂ CONH

And each represents a different type of substituent. )

The N, N'-ethylene-crosslinked dipeptide and a salt thereof having no protecting group according to claim 2 represented by the formula:な Two kinds of different protecting groups may be present-amino acid [NH ₂ — CH (R _t ) -COOH and NH ₂ — CH (R ₂ ) -COOH, and R ₂ is Expression: CH ₃ ,-CH (CH ₃ ) _2, one CH ₂ CH (CH ₃ ) ₂ ,

CH (CH NH ₃ ) CH ₂ CH ₃ , one CH ₂ OH,

(CH ₂ ) 3 NH-C-NH ₂ .-CH ₂ HQ>, CH ₂ HQ) H,

 NH

CH (CH ₃ ) OH,-CH ₂ CH ゥ SCHj,-CH ₂ CONH ₂ , CH ₂ CH ₂ CONH ₂ ,-(CH ₂ ) ₄ NH ₂ ,-CH ₂ COOH, CH ₂ CH ₂ COOH,-CH ₂ SH. -CHj-j— NHlsr, one CH

And the substituents each of which may have a different kind of protecting group. And the general formula (Π):

A compound represented by R ₄ CHCH ₂ R _R (Π) (R ₄ and R “are the same or different and each represents a leaving group) is reacted in an aqueous solution under alkaline conditions, and then reacted in the presence of an acid catalyst. The general formula (I) is characterized by carrying out a cyclization reaction and, if necessary, a deprotection reaction.

The method for producing N, Ν'-diethylene cross-linked dipeptides represented by

 The method according to claim 5, wherein the acid catalyst is ρ-toluenesulfonic acid, methanesulfonic acid, benzenesulfonic acid, trifluoroacetic acid, concentrated sulfuric acid, hydrochloric acid, or hydrochloric acid.

 7. The method according to claim 6, wherein the acid catalyst is sulfuric acid.