KR100230541B1 - Inhibitors of cathepsin g and elastase for preventing connective tissue degradation - Google Patents

Abstract

New compounds that chemically bind the inhibitors of the proteases elastase and kadipsin G prevent the degeneration of connective tissue associated with neutrophil derived inflammatory diseases.

Description

[Name of invention]

Inhibitors of Kadipsin G and Elastase to Prevent Connective Tissue Degeneration

Detailed description of the invention

The present invention relates to novel compounds useful for preventing connective tissue degeneration associated with neutrophil related inflammatory diseases.

[Background of invention]

Elastase and kadipsin G in human neutrophils are involved in tissue destruction associated with many inflammatory diseases such as chronic bronchitis, cystic fibrosis, rheumatoid arthritis [H. L. Malech and J. I. Gallin, New Engl. J. Med., 317 (11), 687 (1987). Elastase and kadipsin G both have extensive proteolytic activity against many connective tissue macromolecules, including elastin, fibronectin, collagen, and proteoglycans. The presence of these enzymes may contribute to the pathology of the disease.

Normal plasma contains large amounts of protease inhibitors that regulate various enzymes associated with connective tissue shifts and inflammation. For example, α-1-antiprotease (α-1-PI) is a serine protease inhibitor that blocks the activity of both elastase and kadepsin G at slower rates. α-1-PI has received considerable attention because the decrease in plasma concentration to less than 15% of normal is associated with early onset of emphysema.

In addition to plasma-derived protease inhibitors, secretions, including bronchial, nasal, cervical mucus and semen, are able to inactivate both elastase and capepsin G and play an important role in maintaining epithelial integrity in the presence of inflammatory cell proteases. It contains an endogenous protease inhibitor termed leukocyte proteinase inhibitor (SLPI), which is believed to be. In certain pathologies, α-1-PI and SLPI are inactivated by neutrophil oxidation mechanisms that allow neutrophil proteases to function essentially in the absence of inhibitors. For example, bronchial lavage fluid obtained from a patient suffering from adult respiratory distress syndrome (ARDS) has been found to contain α-1-PI inactivated by active elastase and oxidation.

Neutrophils have a non-oxidative mechanism in addition to the oxidative mechanism to escape inhibition by antiproteases. Neutrophils in patients with chronic granulomatosis can degrade endothelial cell sarcoma in the presence of excess α-1-PI. There is important in vitro evidence that stimulated neutrophils bind tightly to their substrates and effectively release plasma antiprotease from the microenvironment of firm cell-substrate contact. The introduction of multiple neutrophils into the site of inflammation may cause significant damage to tissue due to the proteolysis occurring at this site.

The present inventors have determined the ability of neutrophil lysates, purified elastase and cardidin G, and stimulated neutrophils to degrade cartilage matrix proteoglycans. I concluded. In addition, the inventors have discovered that stimulated neutrophils degrade the cartilage matrix in the presence of plasma antiprotease, indicating that the degradation occurs in the region of the cell protected by plasma between the neutrophils and the substrate. Degradation of the cartilage matrix occurring in the periphery region could only be blocked by inhibiting both elastase and kadipsin G. We have found a class of enzyme inhibitors that inhibit both elastase and kadipsin G and are therefore useful for preventing neutrophil mediated connective tissue degeneration.

[Summary of invention]

Compounds of the general formula or pharmaceutically acceptable salts thereof are useful for preventing cartilage degeneration.

In the formula,

P ₁ is Ala, bAla, Leu, Ile, Val, Nva, bVal, Met, Nle, Gly, or Sar;

P ₁ ′ is Ala, bAla, Leu, Ile, Val, Nva, bVal, Met, Nle, Phe, Try, Try (Me), Ala (3pyr), Ala (4pyr), Trp, or Nal (1);

P ₂ is Pro, Ind, Ala, bAla, Leu, Ile, Val, Nva, bVal, Met, Nle, Phe, Tyr, Tyr (Me), Ala (3py), Ala (4pyr), Trp, or Nal (1 )ego ;

P ₂ ′ is Pro, Ind, Ala, bAla, Leu, Ile, Val, Nva, bVal, Met, Nle, Gly, Sar or absent;

P ₃ is Lys, Arg, Pro, Ind, Ala, bAla, Leu, Ile, Val, bVal, Met, or Nle;

P ₃ ′ is or absent Ala, bAla, Leu, Ile, Val, Nva, bVal, Met, Nle, Gly, Sar;

P ₄ is Ala, bAla, Leu, Ile, Val, Nva, bVal, Met, Nle or absent;

P ₄ ′ is or absent Ala, bAla, Leu, Ile, Val, Nva, bVal, Met, Nle, Gly, Sar;

L is the following formula

(Wherein n is 0 or an integer of 1 to 6;

p and q are each independently integers of 1 to 6;

r is 1 or 2;

L ₁ and L ₂ are each independently selected from a carbonyl or sulfonyl group, L ₁ is bound to an elastase inhibitory fragment, and L ₂ is bound to a cardipsin G inhibitory fragment;

Ph, Ph ₁ and Ph ₂ are each independently m-phenylene or p-phenylene group;

B is one of a group of a bond,-(CH ₂ ) m-, or -S (O) ₂ N (H) C (O)-,

EIM and CGIM are each independently -C (O) C (O) R, -CF ₂ CF ₃ , -CF ₃ , -CF ₂ H, -CO ₂ R ₃ , -CONHR ₃ , -CF ₂ CHR ₃ C ( O) NHR, -H, alkyl, aryl, aralkyl, -C (O) R, wherein R ₃ is H, alkyl, phenyl, benzyl and R is OH or alkoxy.

Detailed description of the invention

Isosteres of the compounds of formula (I) are (a) (if natural) non-natural sequences of one or more of the α-amino acid residues of the R ₁ substituent, or (b) normal peptide amide bonds are modified, For example -CH ₂ NH- (reduced state), -COCH _2- (keto), -CH (OH) CH _2- (hydroxy), -CH (NH ₂ ) CH _2- (amino), -CH ₂ CH _2- (hydrocarbons). Preferably, the compounds of the present invention should not be present in isoploid form, in particular, free of modified peptide amide groups in R ₁ valence, and even if desired, it is desirable to keep isotope modifications to a minimum.

Unless stated otherwise, it is preferred that the α-amino acid building blocks of these peptidase substrate homologs are L-arrays thereof. As in the conventional nomenclature used by peptide chemists, the amino acid code for which the first (or other) letter of the code is uppercase indicates that the amino acid is a natural "L" sequence, and if the first (or other) letter of the code is lowercase, the amino acid is " D "array. Reference is made herein to lowercase amino acid codes or codes beginning with "(D)-", which are considered to be the same.

These compounds of the present invention having aspartic acid or glutamic acid moieties may be in free form or in salt form such as, for example, acid addition salts or anionic salts. These compounds can be converted from their salt form to the base form or from the base form to the salt form by known techniques. Preferred salts are trifluoroacetic acid salts, hydrochloride salts, sodium, potassium or ammonium salts, but the scope of salts included in the present invention is not limited thereto and the range includes all known salts used in peptide chemistry techniques. Can be extended.

Before defining and / or exemplifying the scope of peptidase inhibitors included in Formula (I), it may be convenient to describe more basic concepts related to peptides. Each α-amino acid has a characteristic “R-group”, which is a side chain or moiety bound to the α-carbon atom of the α-amino acid. For example, the R-group side chain in glycine is hydrogen, methyl in alanine and isopropyl in valine (thus the R ₂ moiety here is the R-group in each α-amino acid presented). For specific R-groups (or side chains) of α-amino acids, it may be helpful to consult AL Lehninger's book, Biochemistry 9 especially chapter 4).

Compounds of formula (I) wherein EIM or CGIM is a —C (O) C (O) R group may exist in hydrated or unhydrated form. Hydrates of triketo compounds having the general formula of (I ') are more chemically effective than non-hydrated triketo compounds of the general formula (I) wherein EIM and / or CGIM are a -C (O) C (O) R group Much more stable.

For this reason, hydrates are preferred, and references to triketo compounds in this specification and in the claims should be considered to include references to the corresponding hydrated forms according to the context. Moreover, the compounds of the present invention are expected to exist in hydrated form under normal physiological conditions.

Recognized abbreviations for α-amino acids are listed in Table 1 below.

TABLE 1a

TABLE 1b

In the present invention, a compound of the formula (1) wherein P ₁ is norvaline or valine is preferred. And P ₁ is norvaline or valine; P ₁ ′ is phenylalanine; P ₂ is proline; P ₂ ′ is proline; P ₃ is isoleucine, valine or alanine; P ₃ ′ is alanine, valine or absent; P ₄ is alanine or absent; Preference is given to compounds of the general formula 1 in which P ₄ ′ is alanine or absent. Preference is given to compounds wherein L is a -C (O) -phenylene-C (O)-group, in particular phenylene is a para-phenylene group. As the compound of the present inventors, a compound of the general formula 1 in which CGIM and EIM are a -CF ₃ or -CF ₂ CF ₃ group is preferable. In particular, -P ₄ -P ₃ -P ₂ -P _1- (SEQ ID NO: 2) is Ala-Ala-Pro-Val- (SEQ ID NO: 3); -Lys (2CBz) -Pro-Val-; Or a -Val-Pro-Val- group is preferred. Also, in particular, -P ₄ '-P ₃ ' -P ₂ '-P ₁ '-(SEQ ID NO: 4) is -Ala-Ala-Pro-Phe- (SEQ ID NO: 5); -Val-Pro-Phe-; Or -Phe-. Most preferred compounds of the present invention include compounds of the structure

Peptidase substrates of formula (I) are used to prevent connective tissue degeneration, such as cartilage degeneration associated with neutrophil-related inflammatory diseases, and have an effective anti-inflammatory effect in the treatment of gout, rheumatoid arthritis and other inflammatory diseases, and elastin-mediated tissue It can be used to treat emphysema and adult respiratory distress syndrome (ARDS) by preventing damage. In the end use of these compounds, the enzyme inhibition properties of the compounds of formula (I) are readily identified by standard biochemical techniques well known in the art. The potential dosage range for the end use of these compounds will of course depend on the nature and severity of the disease as determined by the attending physician, with 0.01 to 10 mg / kg body weight per day useful for the disease state and preferably 0.1 to 10 mg / kg body weight per day.

Although the range of compounds included in general formula (I) is defined, the method by which these compounds are prepared is as follows. Compounds of formula (I) may be prepared using standard chemical reactions similarly known to be useful for preparing a variety of known peptides. A preferred method of preparing a compound of the present invention is to first prepare fragments corresponding to an elastase inhibitory peptide of formula 2 and a kadepsin G inhibitory peptide of formula 3, and then link the two fragments with an "L" group. will be.

_{Wherein, P 1, P 1 ',} P 2, P 2', P 3, P 3 ', P 4, P 4', EIM and CGIM are as defined in general formula 1. The elastase inhibitory peptides of formula 2 and the kadepsin G inhibitory peptides of formula 3 are generally prepared first of the compounds of formulas 4 and 5 below or their protective or activated derivatives, followed by Prepared by addition of the desired amino acid using standard techniques known to those skilled in the art.

In formula, P <_1> , P <_1>', EIM and CGIM are as defined in General formula (1). For this purpose, suitable references for these techniques are available in M. M. Bodanszky and A. The 1985 edition of The Practice of Peptide Synthesis, by A. Bodanszky, includes the use and elimination of protecting groups for each of the amino acids and populations and the variables and techniques that influence selection. This is described in detail, and activation and binding techniques and other special procedures are also included. However, prior to applying these peptide chemistry techniques, certain key intermediates comprising the elastase inhibitory moiety (EIM) and the kadepsin G inhibitory moiety (CGIM) must first be prepared. The preparation of the key intermediate is as follows.

These compounds of formula (2) or (3) can be prepared analogously using standard chemical reactions known in the art. More specifically, the EIM or CGIM is -CF ₂ H, -CF ₃ , -CO ₂ R ₃ , -CO ₂ R ₃ , -CONHR ₃ , -C (O) R, -CF ₂ CHR ₃ C (O) Compounds of formulas 2 and 3 are known which are NHR, H, alkyl, aryl or aralkyl. Thus, the details for preparing compounds of formula 2 or 3 wherein EIM or CGIM is -CF ₂ H, -CF ₃ , -CO ₂ R ₃ , -CO ₂ R ₃ , -CONHR ₃ , or -C (O) R The method can be found in European Patent Application No. 195,212, published September 24, 1986. Detailed methods for preparing compounds of formula 2 or 3 wherein EIM or CGIM are —CF ₂ CHR ₃ C (O) NHR are described in European Patent Application No. 275,101 published July 20, 1988. Detailed methods for preparing compounds of Formulas 2 and 3 wherein EIM and CGIM are H, alkyl, aryl, or aralkyl are described in European Patent Application No. 363,284 published April 11, 1990.

A method for preparing a compound of Formula 2 wherein EIM or CGIM is -C (O) C (O) R is shown in Scheme A, wherein R, P ₁ , P ₂ , P ₃ and P ₄ are as defined above. same. Compounds of formula 3 can be prepared in a similar manner. Specifically, the compound of formula 2 may be prepared by treating a suitable lide of formula 6 with (a) ozone and dimethylsulfide or (b) singlet oxygen. The ozone addition decomposition reaction can be conveniently carried out, for example, by generating an excess of ozone in a suitable solution of a cooling solution of general formula (6). Suitable solvents include all non-reactive solvents in which the illide of Scheme 6 is soluble, for example alkyl esters of simple alkanoic acids such as ethyl acetate; Chlorinated hydrocarbons such as carbon tetrachloride, chloroform, 1,2-dichloroethane, 1,1,2,2-tetrachloroethane, and methylene chloride; Aromatic hydrocarbons such as benzene, toluene, and xylene; Chlorinated aromatic compounds such as 1,2,4-trichlorobenzene and o-dichlorobenzene; Alcohols such as methanol, ethanol and isopropanol; Or ether solvents such as diethyl ether, tetrahydrofuran (THF), and 1,4-dioxane. Methylene chloride is preferred.

Scheme A

The temperature of the ozonation decomposition reaction mixture may be any temperature at which the reaction can be carried out, typically from about -78 ° C to about 0 ° C, preferably from -78 ° C to about -35 ° C, most preferably about- 70 ° C. The reaction time will vary depending on the yield, the concentration of the reactants, the temperature and other factors. For convenience, ozone bubbles were generated in the reaction mixture until the solution turned blue indicating excess ozone.

The ozonate was then treated with a reducing excess of metal such as metal zinc or preferably dimethylsulfide. Preferred compounds of formula 2, such as hydrates, are isolated from the reaction mixture by conventional methods, typically by solvent removal (by evaporation). Purification can be carried out, for example, by flash chromatography.

Oxidation methods using singlet oxygen are well known. More specifically, the method for preparing tricarbonyl esters by oxidizing the illide with singlet oxygen is described by H. Wasserman et al. Amer, Chem. Soc. 11, 371 (1989).

Singlet oxygen can be generated by pigment sensitization excitation of oxygen. Suitable pigments include Rose Bengal, Eosin Y and Methylene Blue. Another photosensitizer is dinaphthalenethiophene. Typically, Rose Bengal and Eosin Y are attached to basic anion exchange resins, and methylene blue is attached to acidic cation exchange resins. This is done by a UV lamp such as a tungsten-iodine lamp. Suitable solvents are all solvents which promote and do not interfere with the desired reaction. These solvents include aromatic hydrocarbons such as benzene and toluene; Hydrocarbons such as hexane; Ether solvents such as diethyl ether, tetrahydrofuran (THF) and 1,4-dioxane; Chlorinated hydrocarbons such as dichloromethane and chloroform; Carbon disulfide; And alcohols such as methanol, ethanol, propanol, isopropanol and t-butanol. These can be mixed and used. The temperature of the reaction mixture may be about -78 ° C to about 30 ° C, typically about -78 ° C to about -50 ° C. The reaction time will vary depending on the reactant, solvent, concentration and temperature and can be from about 1 minute to about 2 hours. Purification and isolation can be accomplished by isolating the product from the process described in the detailed description above and the ozonation decomposition reaction mixture.

Ilide of general formula (6) is prepared from a suitable N-protected illide, preferably phthaloyl protected illi of general formula (7). Removal of the phthaloyl group can be easily carried out by methods generally known to those skilled in the art. For example, the phthaloyl lide solution may be reacted with hydrazine hydrate, typically 20 times excess of hydrazine hydrate, until the reaction is substantially complete. The solvent may be any of those described above for the ozonation decomposition reaction, preferably an alcoholic solvent such as EtOH. The reaction temperature of the mixture is about 0 ° C to about 60 ° C, for convenience, about room temperature, i.e. 25 ° C. The reaction time will vary depending on the particular reactant, temperature, solvent, and other known factors that affect the reaction time. For convenience, the progress of the reaction can be checked by thin layer chromatography (TLC).

Following the removal of the phthaloyl group, P ₂ , P ₃ and P ₄ groups can be bound to amino groups which are currently free. P ₂ , P ₃ and P ₄ can be bound to unprotected free amino compounds by well known peptide binding techniques.

In the binding of each amino acid or peptide to the deprotected compound of formula 7, suitable side chain protecting groups are used. Selection and use of suitable protecting groups for these side chain functional groups will be possible to one skilled in the art and will depend on the presence of other protected amino acid residues in the amine groups and peptides to be protected. The choice of such side chain protecting groups is important in that they should not be removed during the deprotection and binding steps of the synthesis. For example, when Boc is used as the α-amino protecting group, the following side chain protecting group is suitable. the p-toluenesulfonyl moiety can be used to protect the amino side chains of amino acids such as Lys and Arg; p-methylbenzyl, acetamidomethyl, benzyl (Bzl), or t-butylsulfonyl moieties can be used to protect the sulfide containing side chains of amino acids such as cysteine, homocysteine, penicylamine, or derivatives thereof; Benzyl (Bzl) or cyclohexylester moieties can be used to protect the carboxylic acid side chains of amino acids such as Asp or Glu; Benzyl (Bzl) ethers can be used to protect the hydroxy containing side chains of amino acids such as Ser and Thr; The 2-bromocarbobenzoxyl (Z (Br)) moiety can be used to protect the hydroxy containing side chains of amino acids such as Tyr. These side chain protecting groups can be attached and removed according to standard techniques and methods known in the art. These side chain protecting groups are preferably deprotected with anhydrous hydrogen fluoride (1:10) in which anisole is dissolved. Typically deprotection of the side chain protecting groups is carried out after the peptide chain synthesis is completed, but alternatively these can be removed at other appropriate times. When a solid phase synthesis method is used, deprotection of these side chains is preferably performed simultaneously with cleavage of the peptide from the resin.

Phthaloyl illi of Formula 7 is prepared by reacting phthaloyl protected acid chlorides of Formula 8 with phosphonium lide of Formula 9. This reaction is carried out by adding a solution of a suitable lide of general formula (9) to the solution of acid chlorides of general formula (8), preferably dropwise. Suitable solvents include those solvents described above for the ozonation decomposition reaction, with ether solvents such as THF being preferred. The reaction can range from about 30 minutes to about 12 hours, typically from about 2 to about depending on the acid chloride, the lead, the solvent (s) and the temperature (which may be from about 0 ° C. to about 60 ° C., and conveniently at room temperature, ie 25 ° C.). It will take 3 hours. Isolation and purification are carried out by filtration of the reaction mixture to remove solids, followed by chromatography of the filtrate with, for example, a 50% mixture of ethyl acetate and hexane as eluent on silica gel.

Phosphorus Ilide of Formula 9, or Wittig reagent, is prepared in a conventional manner from the corresponding α-halocarboxylic acid derivatives of Formula 10, i.e. Prepared by reaction with a rapid phosphine to obtain a phosphonium salt. When treated with a strong base such as an organolithium compound such as lithium diisopropylamide (LDA), sodium hydride, or sodium amide, the acidic protons are removed to form the desired illide. Suitable solvents used to form the Vitis reagents include, for example, aromatic hydrocarbons such as benzene or toluene, chlorinated hydrocarbons such as carbon tetrachloride, chloroform, or methylene chloride, or ether-based solvents such as diethyl ether or THF. Non-reactive solvents are included.

For convenience, the reaction can be carried out at about 0 ° C to about 60 ° C, typically at room temperature, ie about 25 ° C. The halo group of the α-haloester is preferably a bromine group, but may also be a chloro or iodine group or any good leaving group which forms a stable phosphonium salt such as a mesylate or tosylate group.

The acid chloride of the general formula (8) can be prepared, for example, by reacting the corresponding acid of the general formula (11) with α, α-dichloromethyl methyl ether at reflux. After about 3 hours, the solution was cooled and the product concentrated by evaporating the solvent. The crude acid chloride obtained can be used directly for reaction with phosphorus lide of general formula 9 without further purification.

A method for preparing a compound of Formula 2 wherein EIM or CGIM is -CF ₂ CF ₃ is shown in Scheme B, where R ₁ and R ₂ are as defined above, Pg is carbamate, preferably benzyl Amino protecting groups such as oxycarbonyl (Cbz) groups.

Scheme B

Compounds of formula 3 can be prepared in a similar manner.

In particular, the compounds of the present invention are prepared by reducing N-methoxy-N-methylamides of Formula 15a or 15b to produce aldehydes of Formulas 14a and 14b, respectively. It is preferable to use the compound of general formula 15a as an initial release material. The reduction can be carried out by generally known methods and can be easily carried out by those skilled in the art using lithium aluminum hydride (LAH). This reduction method is conveniently carried out by adding an excess of LAH to a solution of a compound of formula 15a or 15b, typically cooled to about 0 ° C., dissolved in an unreactive solvent such as an ether solvent such as tetrahydrofuran (THF). can do. After the reaction is substantially complete, typically after about 30 minutes, the reaction mixture is quenched, for example by adding 10% potassium hydrogen sulfate followed by water. The product can then be isolated by, for example, extracting the aqueous mixture with a solvent such as telacetate, drying and removing the solvent. The crude product can be purified, for example, by column chromatography or recrystallization such as silica gel column using 55% ethyl acetate / hexane as eluent.

The aldehydes of formulas 14a and 14b are then reacted with pentafluoroethyl anions such as lithium salts of pentafluoroethyl anions to obtain the alcohols of formula 13a or 13b, respectively. This condensation reaction is described in P.G. Gassman and Neil J. O'Reilly, J. Org. Chem. 1987, 52, 2481-2490 can be conveniently carried out by those skilled in the art by modifying the method described in the manual. In this method, perfluoroethyl anions are generated in the reaction system by adding methyllithium / lithium bromide complexes to a solution of aldehyde and pentafluoroethyl iodide in a non-reactive solvent such as diethyl ether. The cooled (-78 ° C. to 0 ° C.) reaction mixture is stirred for about 30 minutes to about 1 hour or until the reaction is substantially complete and then the mixture is poured into excess dilute hydrochloric acid and quenched. The product is isolated by, for example, extracting with diethyl ether and then removing the solvent. The crude product is purified, for example by chromatography on silica gel.

The alcohol of formula 13a or 13b is then oxidized to obtain the amino-protected pentafluoroethylketone of formula 12 or the desired compound of formula 2, respectively. By the known Swern oxidation method, or pyridinium dichromate, or chromium anhydride-pyridinium complex, or des-martin periodinan, ie 1,1,1-tris (acetyloxy) -1,1-dihydro Oxidation can also be carried out by a modified Jones oxidation method using -1,2-benzoiodoxol-3 (1H) -one. Of course, if there is a protecting group on the residue of the α-amino acid constituent block, such protecting group can be removed after oxidation. The bonding process can be carried out according to standard methods known in the art.

Swen oxidation is generally carried out by reacting about 2 to 10 equivalents of dimethylsulfoxide (DMSO) with about 1 to 6 equivalents of trifluoroacetic anhydride [(CF ₃ CO) ₂ O] or oxalyl chloride [(COCl) ₂ ] The reactants are dissolved in an inert solvent, such as methylene chloride (CH ₂ Cl ₂ ), and the reaction is carried out at about −80 ° C. to −50 ° C. under anhydrous conditions under an inert atmosphere (eg, nitrogen or a gas having equivalent function). Proceeds at a temperature of to form a sulfonium addition product in the reaction system, to which about 1 equivalent of a suitable alcohol of formula 13a or 13b is added. Preferably, the alcohol is dissolved in an inert solvent (eg CH ₂ Cl ₂ or a minimum amount of DMSO), the reaction mixture is heated to about −50 ° C. (for about 10-20 minutes) and then tertiary amine (eg, About 3 to 10 equivalents of triethylamine, N-methylmorpholine, etc.) are added to complete the reaction.

In general, the modified Jones oxidation method reacts with an alcohol pyridinium dichromate of the general formula 13a or 13b to bring the reactants together in a water trapping molecular sieve powder (e.g., 3 'molecular sieve powder), the contact being about 0 ° C to 50 C, preferably at room temperature, in the presence of glacial acetic acid, followed by isolation after removal of the amine protecting group.

Alternatively, chromium anhydride-pyridine complexes (ie, Sarset reagents prepared in the reaction system (Fieser and Fieser "Reagents for Organic Synthesis" Vol. 1, pp. 145 and Sarett, et al., JACS 25, 422) (1953)), wherein the complex is prepared in an inert solvent (eg CH ₂ Cl ₂ ) under anhydrous conditions in an inert atmosphere at 0 ° C. to 50 ° C.] to 1 to 5 equivalents of 1 equivalent of alcohol of formula 13a or 13b. The reaction is reacted for about 1-15 hours by addition, and then the amine protecting groups are isolated and optionally removed.

Another method of converting the alcohols of formula 13a or 13b to the preferred ketones of formulas 1 or 5 is the oxidation reaction using des-martin periodinan [Dess and Martim. J. Org. Chem. 48, 4155, (1983). This oxidation method is carried out by contacting a suitable alcohol of the general formula 13a or 13b with 1 to 5 equivalents (preferably 1.5 equivalents) of periodinane, and the reagent is subjected to 0 ° C. to anhydrous conditions under an inert atmosphere (preferably nitrogen). It is a suspension suspended in an inert solvent (eg methylene chloride) at 5 ° C. (preferably room temperature) and the reaction is allowed to react for about 1 to 48 hours. Any deprotection of the amine protecting group may be preferably performed after isolating the ketone.

In one method of preparing the compounds of the invention, the compounds of formula are prepared by first converting the perfluoroethyl alcohol of amino-protected formula 13a to the corresponding compound of formula 13b prior to final oxidation. The amino protected perfluoroethyl alcohol of formula 13a is first deprotected if necessary, and then the amino acid or peptide chain represented by P ₄ -P ₃ -P ₂ -is subjected to standard α-amino acid or peptide binding methods. Can be added. Wherein the P ₄ -P ₃ -P ₂ -group consists of one or more amino acids and the entire peptide chain can be added to the deprotected general formula 13a compound or the amino acids are sequentially linked to the deprotected compound of general formula 13a You can. Alternatively, these two binding methods can be used in combination. In a similar manner the compound of formula 12 can be converted to the desired compound of formula 2.

Suitable side chain protecting groups are used in binding each amino acid or peptide to the deprotected compound of Formula 13a or Formula 12. Selection and use of suitable protecting groups for these side chain functional groups will be possible to one skilled in the art and will depend on the presence of protected amino acids and other protected amino acid residues in the peptide. It is important to note that the choice of such side chain protecting groups should not be removed during the deprotection and binding steps of the synthesis. For example, when Boc is used as the α-amino protecting group, the following side chain protecting group is suitable. the p-toluenesulfonyl moiety can be used to protect the amine side chains of amino acids such as Lys and Arg; p-methylbenzyl, acetamidomethyl, benzyl (Bzl), or t-butylsulfonyl moieties can be used to protect the sulfide containing side chains of amino acids such as cysteine, homocysteine, penicylamine, or derivatives thereof; Benzyl (Bzl) or cyclohexylester moieties can be used to protect the carboxylic acid side chains of amino acids such as Asp or Glu; Benzyl (Bzl) ethers can be used to protect the hydroxy containing side chains of amino acids such as Ser and Thr; The 2-bromocarbobenzoxoxy (2Br-Z) moiety can be used to protect the hydroxy containing side chains of amino acids such as Tyr. These side chain protecting groups can be attached and deposited according to standard techniques and methods known in the art. These side chain protecting groups are preferably deprotected with anhydrous dissolved hydrogen fluoride (1:10). Typically deprotection of the side chain protecting groups is performed after the peptide chain synthesis is completed, but alternatively these can be removed at other appropriate times. When a solid phase synthesis method is used, deprotection of these side chains is preferably performed simultaneously with cleavage of the peptide from the resin.

As a preferred example of the preparation of the compounds of the present invention, the compounds of the general formulas 15a and 15b can be used as the compounds of the general formulas 15a and 15b by condensing N-methoxy-N-methylamide with lithium salts of perfluoroethyl anions. It can be converted directly to each of the compounds of formula 12 or 2 in the same manner as the conversion to each compound of Formulas 14a and 14b.

The compound is then isolated and purified by standard techniques. Preferred amino acids, derivatives and isomers thereof can be obtained as a commodity or synthesized according to standard techniques and methods known in the art.

The N-methoxy-N-methyl amides of Formulas 15a and 15b are as defined above for which R ₁ and R ₂ are defined for Formula 1, respectively, and Pg is carbamate, preferably benzyloxycarbonyl (Cbz) group. From the corresponding α-amino acids of the general formulas 16a and 16b which are the same amino protecting groups (see JA, Fehrentz and B. Castro, Synthesis, 676-78 (1983)).

Isobutylchloroformate is cooled (ie, -60 ° C. to about 0 ° C.) of N-methylmorpholine or another stericly masked non-nucleophilic tertiary amine and α-amino acid compound in a non-reactive solvent such as methylene chloride. To the mixture. After about 5 minutes to about 1 hour, typically about 15 to 20 minutes, NO-dimethylhydroxylamine HC1 is added and the mixture is stirred for about 30 minutes up to about 6 hours, and then the reaction mixture is allowed to warm to room temperature. . When the reaction is substantially complete, typically, after about 1 to about 10 hours, the mixture is poured into water and the aqueous layer is extracted, for example with ethyl acetate. The desired compound can then be isolated by evaporation of the solvent and purification can be carried out, for example, by flash chromatography on silica gel with ethyl acetate / hexane as the eluent. Purification can be carried out, for example, by flash chromatography on silica gel using methylene chloride as the eluent.

Compounds of Formula 1 wherein L ₁ and L ₂ are both carbonyl groups, L ₁ and L ₂ are both sulfonyl groups or L ₂ is a carbonyl group, and L ₁ is a sulfonyl group can be prepared by techniques and methods well known to those skilled in the art. have. General synthetic schemes for preparing these compounds of Formula 1 are described in Scheme C. In Scheme C, unless otherwise stated, all substituents are as previously defined.

Scheme C

Scheme C is a general synthetic method for preparing compounds of formula (I), wherein L ₁ and L ₂ are both carbonyl groups, L ₁ and L ₂ are both sulfonyl groups, or L ₂ is a carbonyl group, and L ₁ is a sulfonyl group. To provide.

In step a, a suitable elastase inhibitory peptide fragment of formula (2) is combined with a suitable derivative of L represented by formula (17) by known techniques to yield the corresponding L-elastase of formula (18) Provide inhibitory peptide fragments.

When the compound of formula (1) is L ₁ and L ₂ are both carbonyl groups, a suitable derivative of L represented by formula (17) is a carboxylic acid in which the L ₂ carbonyl group is t-butyloxycarbonyl protected, and L ₁ And a carbonyl group is an unprotected carboxylic acid.

When the compounds of formula (1) are L ₁ and L ₂ are both sulfonyl groups, suitable derivatives of L represented by formula (17) are sulfonic acids in which the L ₁ sulfonyl group is a sulfonyl chloride group and the L ₂ sulfonyl group is unprotected Phosphorus compound.

When the compound of formula (1) is L ₂ is a carbonyl group and L ₁ is a sulfonyl group, a suitable derivative of L represented by formula (17) is a carboxylic acid in which the L ₂ carbonyl group is t-butyloxycarbonyl protected, L ₁ sulfonyl group is a sulfonyl chloride group.

In step b, a suitable L-elastase inhibitory peptide fragment of formula (18) is combined with a suitable cardipsin G inhibitor peptide fragment of formula (3) by techniques known in the art to Obtain the corresponding compound.

Suitable L-elastase inhibitory peptide fragments of Formula (18) are first hydrolyzed by techniques known in the art prior to the binding reaction in step b, wherein the carboxylic acid with L ₂ carbonyl group t-butyloxycarbonyl protected Should be.

Starting materials used in Scheme C are readily available to those of ordinary skill in the art.

Compounds of formula (1) wherein L ₁ is a carbonyl group and L ₂ is a sulfonyl group can be prepared by techniques and methods well known to those skilled in the art. General synthetic schemes for preparing these compounds of formula (1) are described in Scheme D. In Scheme D, all substituents are as described above unless otherwise indicated.

Scheme D

Scheme D provides a general synthetic process for preparing compounds of formula (1) wherein L ₁ is a carbonyl group and L ₂ is a sulfonyl group.

In step a, a suitable kadepsin G inhibitor peptide fragment of general formula (3) is combined with a suitable L derivative represented by general formula (19) by a technique known in the art to correspond to the corresponding cardidine of general formula (20) Obtain a G inhibitory peptide fragment.

Suitable derivatives represented by formula (19) are compounds in which the L ₁ carbonyl group is t-butyloxycarbonyl protected carboxylic acid and the L ₂ carbonyl group is a sulfonyl chloride group.

In step b, a suitable capepsin G inhibitor peptide fragment of formula (20) is combined with a suitable elastase inhibitor peptide fragment of formula (2) by a technique known in the art to correspond to the corresponding Obtain the compound.

The t-butyloxycarbonyl protected carboxylic acid functional group on L ₁ of a suitable cardipine G inhibitory peptide fragment of formula (20) must first be hydrolyzed by techniques known in the art prior to the binding reaction in step b. do.

Starting materials for use in Scheme D are readily available in the art.

The following specific examples are intended to illustrate the preparation of the present invention and are not intended to limit the scope of the compound to the range of compounds including Formula (I).

Example 1

[[CF ₃ ] Phe-Pro-Val-C (O) -phenylene-C (O) -Val-Pro-Val [CF ₂ CF ₃ ] -S-EQ ID NO: 6 Boc-Val [CF ₂ CF ₃ ] of manufacture]

1.0 g (3.8 mmol) of Boc-Val dimethylhydroxyamide was dissolved in 50 ml of ethyl ether and cooled to -78 ° C. 3 g (12.2 mmol) of pentafluoroethyl iodide were added followed by 6 ml of methyllithium lithium bromide complex (1.5 M solution). 3 g (12.2 mmol) of pentafluoroethyl iodide was added followed by the addition of 6 ml of methyllithium lithium bromide complex (1.5 M solution) three times. Stir at −78 ° C. for 15 minutes, then warm to room temperature. The reaction was poured into water and the organic phase separated. The aqueous layer was extracted with ethyl ether (150 ml x 3 times) and the organic layers combined and dried (using Na ₂ SO ₄ ). The solvent was evaporated in vacuo and purified by silica gel chromatography (10% ethyl acetate / hexanes) to afford the title compound.

[Production of Val [CF ₂ CF ₃ ] · Hydrochloride

350 g (1.1 mmol) of Boc-Val [CF ₂ CF ₃ ] was dissolved in 50 ml of ethyl ether and cooled to 0 ° C. Treated with hydrogen chloride gas for 5 minutes and stirred for 30 minutes. The solvent was removed in vacuo to afford the title compound.

[Production of Boc-Val-Pro-Val [CF ₂ CF ₃ ]]

314 mg (1.0 mmol) of Boc-Val-Pro was dissolved in 4 ml of methylene chloride and 252 mg (2.5 mmol) of N-methylmorpholine were added. The mixture was cooled to -22 ° C and 136 mg (1.0 mmol) of isobutylchloroformate were added. Stir for 20 minutes and add to Val [CF ₂ CF ₃ ]. Hydrochloride (1.1 mmol). Stir at −22 ° C. for 1 h, warm to rt and stir for 3 h. Purification by silica gel chromatography (40% ethyl acetate / hexanes) gave 405 mg of the title compound.

Val-Pro-Val [CF ₂ CF ₃ ] Preparation of Hydrochloride

385 mg (0.74 mmol) of Boc-Val-Pro-Val [CF ₂ CF ₃ ] was dissolved in 50 ml of ethyl ether and cooled to 0 ° C. Treated with hydrogen chloride gas for 5 minutes and stirred for 30 minutes. The solvent was removed in vacuo to yield 334 mg of the title compound.

[Production of Boc-Val-Pro-Phe [OH] [CF ₃ ]]

300 g of ² -phenyl-Δ ² -oxazoline-4-phenylmethyl-5-one [see Synthesis, # 3, 191-3 (1982)] and 700 g of trifluoroacetic anhydride were mixed. Heated at reflux for 3 hours and then stirred overnight at room temperature. The solvent was evaporated in vacuo and 400 g of oxalic acid were added. Stir and add 50 g of oxalic acid further. Heating was continued until CO ₂ evolution ceased and a solid formed. Cool to room temperature and dissolve in 12 l of water / ethylacetate 1: 3 mixture. The organic layer was separated and washed until saturated with saturated sodium bicarbonate and then with water. Dry (use MgSO ₄ ), filter, concentrate by boiling to 2.5 l volume. Cool to room temperature and add hexane 11. The precipitated solid was filtered and air dried to afford 187.6 g of 1,1,1-trifluoro-2-one-3-benzoylamino-4-phenylbutane.

187.6 g of 1,1,1-trifluoro-2-one-3-benzoylamino-4-phenylbutane was dissolved in 1 ethanol and cooled in an ice bath. Over 15 minutes, 11 g of sodium borohydride was added to you in portions. The ice bath was removed and stirred at room temperature for 1.5 hours. The ice bath was replaced and carefully treated with 250 ml of 10% hydrochloric acid. 4 l of ethyl acetate was added to dissolve followed by 500 ml of water. The organic layer was separated, washed with brine (300 ml x 4) and dried (using MgSO ₄ ). Filter and evaporate the solvent in vacuo. Hexane was added and filtered to give 145.2 g of 1,1,1-trifluoro-3-benzoylamino-4-phenyl-2-butanol as a white solid.

145.2 g of 1,1,1-trifluoro-3-benzoylamino-4-phenyl-2-butanol, 1.4 l of concentrated hydrochloric acid, 700 ml of water and 1 l of ethanol were mixed. Heat at reflux for 24 h, then add 400 ml of concentrated hydrochloric acid and 1.2 l of ethanol. It was further stirred for 24 hours. Ethanol was evaporated in vacuo and filtered. The filtrate was cooled to room temperature and treated with sodium hydrogen carbonate followed by 50% sodium hydroxide while cooling in an ice bath. When the pH reached 10, the solid was filtered off to obtain 58.2 g of [CF ₃ ] [OH] -Phe.

3.3 g (10.5 mmol) of Boc-Val-Pro were dissolved in 25 ml of methylene chloride and 2.12 g (21 mmol) of N-methylmorpholine were added. Cool to −22 ° C. and add 1.43 g (10.5 mmol) of isobutylchloroformate. After stirring at −22 ° C. for 25 minutes 2.5 g (11.5 mmol) of [CF ₃ ] [OH] -Phe were added. Stir at −22 ° C. for 3 hours, warm to room temperature, and stir overnight. The mixture was poured into 100 ml of water and extracted with ethyl ether (150 ml x 3 times). The combined organic layers were washed with dilute hydrochloric acid followed by saturated sodium hydrogen carbonate. Dry (use Na ₂ SO ₄ ) and evaporate the solvent in vacuo. Purification by silica gel chromatography (40% ethyl acetate / hexanes) gave 5.27 g of the title compound.

[Production of Boc-Val-Pro-Phe [CF ₃ ]]

0.79 g (1.54 mmol) of Boc-Val-Pro-Phe [OH] [CF ₃ ] was dissolved in 25 ml of methylene chloride, and 2.5 g of Dess-Martin reagent was added. After stirring overnight at room temperature, 50 ml of water containing 1.0 g of sodium bicarbonate and 1.7 g of sodium hydrogen sulfate were poured. Extracted with ethyl ether (100 ml x 3 times) and dried (using Na ₂ SO ₄ ). The solvent was evaporated in vacuo and purified by silica gel chromatography (40% ethyl acetate / hexane) to give 755 mg of the title compound.

[Production of Val-Pro-Phe [CF ₃ ] · Hydrochloride

Boc-Val-Pro-Phe [CF ₃ ] was dissolved in 100 ml of ethyl acetate and cooled to 0 ° C. Treated with hydrogen chloride gas for 5 minutes and stirred at 0 ° C. for 30 minutes. Evaporation of the solvent in vacuo gave 840 mg of the title compound.

[Production of Boc-phenylene-C (O) -Val-Pro-Phe [CF ₃ ]]

370 mg (1.67 mmol) of Boc-phenylene-C (O) OH was dissolved in 4 ml of methylene chloride and 0.18 ml (1.67 mmol) of N-methylmorpholine. Cool to −20 ° C., add 0.227 g (1.79 mmol) of isobutylchloroformate and stir for 45 minutes. 2 ml of methylene chloride were added, and 0.18 ml of N-methylmorpholine and 0.75 g (1.67 mmol) of Val-Pro-Phe [CF ₃ ] hydrochloride were added. Stir at −20 ° C. for 2 h, warm to rt and stir for 3 h more. This was poured into a mixture of 10 ml of methylene chloride and 20 ml of water. The organic layer was separated and the aqueous layer was extracted with methylene chloride (20 ml x 2). The organic layers were combined and dried (using Na ₂ SO ₄ ). The solvent was evaporated in vacuo and purified by silica gel chromatography (40% ethyl acetate / hexanes) to give 575 mg of the title compound.

[Preparation of H ₂ OC-phenylene-C (O) -Val-Pro-Phe [CF ₃ ] hydrochloride]

250 mg of Boc-phenylene-C (O) -Val-Pro-Phe [CF ₃ ] was dissolved in 50 ml of ethyl acetate and cooled to 0 ° C. Treated with hydrogen chloride gas for 5 minutes and stirred at 0 ° C. for 1 hour. The solvent was evaporated in vacuo to give 232 mg of the title compound.

[[CF ₃ ] Phe-Pro-Val-C (O) -phenylene-C (O) -Val-Pro-Val [CF ₂ CF ₃ ]-Preparation of SEQ ID NO: 6]

Dissolve 230 mg (0.41 mmol) of H ₂ OC-phenylene-C (O) -Val-Pro-Phe [CF ₃ ] hydrochloride in ₃ ml of methylene chloride and 41.4 mg (0.41 mmol) of N-methylmorpholine were dissolved. Added. Cool to −20 ° C. and add 55.7 mg (0.41 mmol) of isobutylchloroformate. Stir at −20 ° C. for 45 min and add 185 mg (0.41 mmol) of Val-Pro-Val [CF ₂ CF ₃ ] .hydrochloride. Stir at −22 ° C. for 3 hours, warm to room temperature, and stir for an additional 3 hours. It was poured into water and extracted with methylene chloride (25 ml x 3 times). The organic layers were combined and dried (using Na ₂ SO ₄ ). The solvent was evaporated in vacuo and purified by silica gel chromatography using ethyl acetate followed by methanol to give 190 mg of the title compound.

Example 2

[[CF ₃ ] Phe-C (O) -phenylene-C (O) -Val-Pro-Val [CF ₂ CF ₃ ]-Preparation of SEQ ID NO: 16]

[Production of Boc-phenylene-C (O) -Phe [OH] [CF ₃ ]]

0.165 g (2.8 mmol) of Boc-phenylene-C (O) OH was dissolved in 6 ml of methylene chloride, and 0.6 g of N-methylmorpholine was added. Cool to −22 ° C. and add 0.4 ml (3.08 mmol) of isobutylchloroformate. Stirred at −22 ° C. for 25 minutes and added a solution of 0.64 g (2.9 mmol) of Phe [OH] [CF ₃ ] in 2 ml of methylene chloride and 0.3 g of N-methylmorpholine. Stir at −22 ° C. for 1 h, warm to rt and stir for a further 2 h. The mixture was poured into 100 ml of water and extracted with ethyl ether (100 ml, then 50 ml). The organic layers were combined and dried (using Na ₂ SO ₄ ). The solvent was evaporated in vacuo and purified by silica gel chromatography (40% ethyl acetate / hexanes) to give 840 mg of the title compound.

[Production of Boc-phenylene-C (O) -Phe [CF ₃ ]]

0.6 g of Boc-phenylene-C (O) -Phe [OH] [CF ₃ ] was dissolved in 25 ml of methylene chloride and 1.8 g of Dess-Martin reagent was added. It was stirred for 48 hours and poured into a mixture of 0.8 g sodium hydrogen carbonate and 1.41 g sodium hydrogen sulfate in 25 ml of water and 100 ml of ethyl ether. The organic layer was separated, and the aqueous layer was extracted with 50 ml of ethyl ether. The organic layers were combined and dried (using MgSO ₄ ). The solvent was evaporated in vacuo and purified by silica gel chromatography (25% ethyl acetate / hexane) to give 430 mg of the title compound.

[Preparation of H ₂ OC-phenylene-C (O) -Phe [CF ₃ ]]

216 mg (0.51 mmol) of Boc-phenylene-C (O) -Phe [CF ₃ ] was dissolved in 50 ml of ethyl acetate and cooled to 0 ° C. Treated with hydrogen chloride gas for 5 minutes and stirred for 3 hours. The solvent was evaporated in vacuo to give 197 mg of the title compound.

[[CF ₃ ] Phe-C (O) -phenylene-C (O) -Val-Pro-Val [CF ₂ CF ₃ ]-Preparation of SEQ ID NO: 16]

175 mg of H ₂ OC-phenylene-C (O) -Phe [CF ₃ ] was suspended in ₂ ml of methylene chloride and 100 μl of N-methylmorpholine was added. Cool to −22 ° C. and add 65 μl isobutylchloroformate. Stir at −22 ° C. for 25 minutes and add a solution of 240 mg of Val-Pro-Val [CF ₂ CF ₃ ] in ₂ ml of methylene chloride and 60 μl of N-methylmorpholine. Stir at −22 ° C. for 30 min, warm to rt and stir for an additional 1.5 h. The solvent was evaporated in vacuo to about 1 ml volume and purified by silica gel chromatography (50% ethyl acetate / hexanes) to give 110 mg of the title compound.

Example 3

[[CF ₃ ] Phe-Pro-Val-C (O) -Val-Pro-Val [CF ₂ CF ₃ ]-SEQ ID NO: 17]

200 mg of Val-Pro-Val [CF ₂ CF ₃ ] hydrochloride was partitioned between 10 ml of ethyl acetate and 20 ml of saturated sodium bicarbonate. The organic layer was separated, and the aqueous layer was extracted with ethyl acetate (20 ml x 3 times). The organic layers were combined, dried (using MgSO ₄ ) and the solvent was evaporated in vacuo to give Val-Pro-Val [CF ₂ CF ₃ ].

200 mg of Val-Pro-Phe [CF ₃ ] hydrochloride was partitioned between 10 ml of ethyl acetate and 20 ml of saturated sodium carbonate. The organic layer was separated and the aqueous layer was extracted with ethyl acetate (20 ml x 3 times). The organic layers were combined, dried (using MgSO ₄ ) and the solvent was evaporated in vacuo to give Val-Pro-Phe [CF ₃ ].

545 mg (5.51 mmol) of phosgene was dissolved in 3.5 ml of benzene, and a solution of 568 mg (1.37 mmol) of Val-Pro-Val [CF ₂ CF ₃ ] in 1 ml of benzene was slowly added. Stirred at 60-75 ° C. for several hours and then vigorously refluxed for two hours. About 1/2 of benzene was evaporated off and the remaining solvent was cooled in an ice bath. A solution of 500 mg (1.37 mmol) of Val-Pro-Phe [CF ₃ ] in 2 ml of ethyl ether was added and stirred for several hours at room temperature. The solvent was evaporated in vacuo and purified by chromatography to give the title compound.

Example 4

[[CF ₃ ] Phe-Pro-Val-CH ₂ -CH ₂ -Val-Pro-Val [CF ₂ CF ₃ ]-SEQ ID NO: 18]

Dissolve 568 mg (1.37 mmol) of Val-Pro-Val [CF ₂ CF ₃ ] and 500 mg (1.37 mmol) of Val-Pro-Phe [CF ₃ ] in 20 ml of methanol, and glyoxal (200 mg of a 40% solution in water, 1.37 mmol). 86 mg (1.37 mmol) of sodium cyanoborohydride and 1 drop of 1% bromocresol green in ethanol were added. The pH of the reaction was maintained with 1 N hydrochloric acid in methanol until the indicator no longer changed. The solvent was evaporated in vacuo and the residue was partitioned in 5 ml of 1 N sodium hydroxide and 10 ml of ethyl acetate. The organic layer was separated, dried (using MgSO ₄ ) and the solvent was evaporated in vacuo. Purification by chromatography gave the title compound.

Sequence list

(1) General Information:

(Ⅰ) Applicant: Angelastro, Michael R

Bay, Philippe

Doherty. Doherty, Niall S

Janusz, Michael J

Mehdi, Shujaath

P. Norton P

(Ii) Name of the Invention: Inhibitors of capepsin G and elastase for preventing connective tissue degeneration

(Iii) Number of sequences: 18

(Iii) communication address:

(A) Address: Marion Merel Dow Inc.

(B) A: East Galbrais Road 2110

(B) City: Cincinnati P. Box 156300

(D) State: Ohio

(E) Country: United States

(F) ZIP Code: 45125-6300

(Iii) computer-readable form:

(A) Medium type: floppy disk

(B) Computer: IBM Compatibility PC

(C) Execution System: PC-DOS / MS-DOS

(D) Software: PatentIn Release # 1.0,

Version # 1.25

(Iii) Current application data:

(A) Application No. US Patent Application No. 07 / 704,449

(B) Application date: May 23, 1991

(C) classification:

(Ⅶ) Patent Attorney / Agent:

(A) Name: Nesbitt, Stephen L

(B) registration number: 28,981

(C) Reference / List No .: M01593

(Iii) Telegraph Call Information:

(A) Telephone Number: (513) 948-7965

(B) Telex: (513) 948-7961

(C) Fax: 214320

(2) Information about SEQ ID NO 1:

(i) Sequence Properties:

(A) Length: 8 amino acids

(B) Type: amino acid

(D) form: linear

(ii) Molecular form: peptide

(xi) the sequence of SEQ ID NO 1

(2) Information about SEQ ID NO 2:

(i) Sequence Properties:

(A) Length: 4 amino acids

(B) Type: amino acid

(D) form: linear

(ii) Molecular form: peptide

(xi) the sequence of SEQ ID NO 2

(2) Information about SEQ ID NO 3:

(i) Sequence Properties:

(A) Length: 4 amino acids

(B) Type: amino acid

(D) form: linear

(ii) Molecular form: peptide

(xi) the sequence of SEQ ID NO 3

(2) Information about SEQ ID NO 4:

(i) Sequence Properties:

(A) Length: 4 amino acids

(B) Type: amino acid

(D) form: linear

(ii) Molecular form: peptide

(xi) the sequence of SEQ ID NO 4

(2) Information about SEQ ID NO 5:

(i) Sequence Properties:

(A) Length: 4 amino acids

(B) Type: amino acid

(D) form: linear

(ii) Molecular form: peptide

(xi) the sequence of SEQ ID NO 5

(2) Information about SEQ ID NO 6:

(i) Sequence Properties:

(A) Length: 6 amino acids

(B) Type: amino acid

(D) form: linear

(ii) Molecular form: peptide

(xi) Features:

(A) Name / Key: Modified Site

(B) location: 1

(D) Other information: / week = "Xaa is a valine homologue with a -CF ₂ CF ₃ group instead of the hydroxyl of the terminal carboxy."

(ix) features

(A) Name / Key: Modified Site

(B) location: 3

(D) Other information: / week = 'Xaa is a valine homologue with L ₁ -Ph-L ₂ -groups instead of one of the hydrogens on the amino terminus. "

(ix) features

(A) Name / Key: Modified Site

(B) location: 3

(D) Other information: / week = "(cont) is carbonyl, each bound to a peptide fragment having a site where L ₁ is modified at position 1-2."

(ix) features

(A) Name / Key: Modified Site

(B) location: 3

(D) Other Information: / Note = "(cont) are each, L ₂ is coupled to a peptide fragment having a deformed portion at 4-6 Ph is p- phenylene group."

(ix) Features:

(A) Name / Key: Modified Site

(B) location: 6

(D) Other information: / week = "Xaa is a phenylalanine homologue with a -CF ₃ group instead of the hydroxyl of the terminal carboxy."

(ix) the sequence of SEQ ID NO 6

(2) Information about SEQ ID NO 7:

(i) Sequence Properties:

(A) Length: 4 amino acids

(B) Type: amino acid

(D) form: linear

(ii) Molecular form: peptide

(xi) the sequence of SEQ ID NO 7

(2) Information about SEQ ID NO 8:

(i) Sequence Properties:

(A) Length: 4 amino acids

(B) Type: amino acid

(D) form: linear

(ii) Molecular form: peptide

(xi) the sequence of SEQ ID NO 8

(2) Information about SEQ ID NO 9:

(i) Sequence Properties:

(A) Length: 4 amino acids

(B) Type: amino acid

(D) form: linear

(ii) Molecular form: peptide

(xi) the sequence of SEQ ID NO 9

(2) Information about SEQ ID NO 10:

(i) Sequence Properties:

(A) Length: 4 amino acids

(B) Type: amino acid

(D) form: linear

(ii) Molecular form: peptide

(xi) the sequence of SEQ ID NO 10

(2) Information about SEQ ID NO 11:

(i) Sequence Properties:

(A) Length: 4 amino acids

(B) Type: amino acid

(D) form: linear

(ii) Molecular form: peptide

(xi) the sequence of SEQ ID NO 11

(2) Information about SEQ ID NO 12:

(i) Sequence Properties:

(A) Length: 4 amino acids

(B) Type: amino acid

(D) form: linear

(ii) Molecular form: peptide

(xi) the sequence of SEQ ID NO 12

(2) Information about SEQ ID NO 13:

(i) Sequence Properties:

(A) Length: 4 amino acids

(B) Type: amino acid

(D) form: linear

(ii) Molecular form: peptide

(xi) the sequence of SEQ ID NO 13

(2) Information about SEQ ID NO 14:

(i) Sequence Properties:

(A) Length: 4 amino acids

(B) Type: amino acid

(D) form: linear

(ii) Molecular form: peptide

(xi) the sequence of SEQ ID NO 14

(2) Information about SEQ ID NO 15:

(i) Sequence Properties:

(A) Length: 4 amino acids

(B) Type: amino acid

(D) form: linear

(ii) Molecular form: peptide

(xi) the sequence of SEQ ID NO 15

(2) Information about SEQ ID NO 16:

(i) Sequence Properties:

(A) Length: 4 amino acids

(B) Type: amino acid

(D) form: linear

(ii) Molecular form: peptide

(ix) Features:

(A) Name / Key: Modified Site

(B) location: 1

(D) Other information: / week = "Xaa is a valine homologue with a -CF ₂ CF ₃ group instead of the hydroxyl of the terminal carboxy."

(ix) features

(A) Name / Key: Modified Site

(B) location: 3

(D) Other information: / week = "Xaa is a valine homologue with L ₁ -Ph-L ₂ -groups instead of one of the hydrogens on the amino terminus."

(ix) features

(A) Name / Key: Modified Site

(B) location: 3

(D) Other information: / week = "(cont) is carbonyl, each bound to a peptide fragment having a site where L ₁ is modified at position 1-2."

(ix) features

(A) Name / Key: Modified Site

(B) location: 3

(D) Other Information: / Note = "(cont) are each, L ₂ is coupled to a peptide fragment having a deformed portion at the 4-position Ph it is p- phenylene group."

(ix) Features:

(A) Name / Key: Modified Site

(B) location: 4

(D) Other information: / week = "Xaa is a phenylalanine homologue with a -CF ₃ group instead of the hydroxyl of the terminal carboxy."

(xi) the sequence of SEQ ID NO 16

(2) Information about SEQ ID NO 17:

(i) Sequence Properties:

(A) Length: 4 amino acids

(B) Type: amino acid

(D) form: linear

(ii) Molecular form: peptide

(ix) Features:

(A) Name / Key: Modified Site

(B) location: 1

(D) Other information: / week = "Xaa is a valine homologue with a -CF ₂ CF ₃ group instead of the hydroxyl of the terminal carboxy."

(ix) features

(A) Name / Key: Modified Site

(B) location: 3

(D) Other information: / week = "Xaa is a phenylalanine homologue with a -CF ₃ group instead of one of the hydrogens on the amino terminus."

(ix) features

(A) Name / Key: Modified Site

(B) location: 6

(D) Other information: / week = "Xaa is a phenylalanine homologue with a -CF ₃ group instead of the hydroxyl of the terminal carboxy."

(xi) the sequence of SEQ ID NO 17

(2) Information about SEQ ID NO 18:

(i) Sequence Properties:

(A) Length: 6 amino acids

(B) Type: amino acid

(D) form: linear

(ii) Molecular form: peptide

(ix) Features:

(A) Name / Key: Modified Site

(B) location: 1

(D) Other information: / week = "Xaa is a valine homologue with a -CF ₂ CF ₃ group instead of the hydroxyl of the terminal carboxy."

(ix) features

(A) Name / Key: Modified Site

(B) location: 3

(D) Other information: / week = "Xaa is a valine homologue with a-(CF ₂ ) _{n + 2-} group where n is 0 instead of one of the hydrogens on the amino terminus."

(ix) features

(A) Name / Key: Modified Site

(B) location: 6

(D) Other information: / week = "Xaa is a phenylalanine homologue with a -CF ₃ group instead of the hydroxyl of the terminal carboxy."

(xi) the sequence of SEQ ID NO 18

Claims (3)

A compound of formula 1 or a pharmaceutically acceptable salt thereof. Wherein P ₁ is Val, P ₁ ′ is Phe, P ₂ is Pro, P ₂ ′ is Pro, P ₃ is Val, P ₃ ′ is Val or absent, P ₄ is absent , P ₄ ′ is Val or absent, and if one of P ₃ ′ and P ₄ ′ is Val, the other is absent and L is —C (O) —Ph—C (O) — or —CH ₂ — CH _2- , EIM is -CF ₂ CF ₃ , and CGIM is -CF ₃ . The compound of claim 1, wherein L is a -C (O) -phenylene-C (O)-group. The compound of claim 1, wherein