CA2109516C - Inhibitors of cathepsin g and elastase for preventing connective tissue degradation - Google Patents

Abstract

Novel compounds which are chemically linked inhibitors of the proteases Elastase and Cathepsin G prevent connective tis-sue degradation associated with neutrophil induced inflammatory disease.

Description

2~.~ ~.

INHIBITORS OF CPrTHEPSIN G AND ELASTASE FOR PREVENTTNG
COi<713ECTIVE TISSUE DEGRADATION
This i.nventi.on ~eela~es to r~ov~l chemical c~mpounds useful for preventing connective tisshe degradation associated with neutrophil associated inflammatory disease.
BACKGROUND OF THE TNVENTION
Human neu~rop~~l elas~ase and cath~psin G have been implicated in the tissue destruct~:on associated with a number of inflammatory diseases such as chronic bronchitis, cystic fibrosis, and rheumatoid arthritis. H. L. Ma~.ech and ~. T. ~allin.'Neca Engl. JMed:; 317(11). 687 (1987).
~5 Both ela~ta~e and cathepsin G'have a broad range of proteolytic-activity .against a nbanber of connedtive ti-ssu~
znacromalecules including elastin, fibronectin, collagen, and proteoglycan~' The presence of these enzymes may contribute to the pathology of these diseases:
Normal plasma contains large-quantities of pro ease inhibitors that control a var~.ety of enzymes involved in ' c~~nect-~ive tissue turnover and inflammation. For exampler a-1-antiprot~ase (Q-1'-PL) is a shrine protease inhibitor 25 that blocks the activity of both elastase and, at a slower':
rate; cathepsin G. a-°1-PI has received considerable interest because reducta.an in plasana l,evel,s to lens than ~1~~5~.~~
veto ~zizo~s7 ~~riu~92io~zs~
_2_ 15~ of normal is associated with the early development of emphysema.
In addition to plasma derived protease inhibitors, secretory fluids, including bronchial, nasal, cervica l mucus, and seminal fluid contain:an,endogenous proteasE
inhibitor called secretory leukoprotease inhibitor (SLPZ) that can inactivate both elastase and cathepsin G and is believed to play an important role in maintaining the integrity of the epithelium in the presence of inflammatory cell proteases. In certain pathcalogical states a-1-PI and SLPI are inactivated by neutrophil oxidative mechanisms allowing the neutrophil prot~ases to function in an essentially inhibitor°free environment. For example;
bronchial lavage fluids from patients with'adult respiratory distress syndrome CARDS) have been found to contain active elastase and a-1°PI that had been inactivated by oxidation.
In addition to oxidative mechanisms; neutrophils possess non--oxiclati ~e mechanism's for eluding inhibition by antiproteases. Neutrophils from patients with chronic granulomatous disease are capable of degrading endothelial cell matrices in the presence of excess a-1-PI. There is considerable in vitro evidenee that stimulated neutrophils can tightly bind to their substrates such that serum antiproteases are effecta.vely excluded from the microenviroment of tight cell-substrate contact. The influx of large numbers of neutrophils to an inflammatory site may result in considerable tissue damage due to the proteo~.~yri.s that occurs in this regions Applicants have determined tha t elastase and cathepsin G are the primary neutrophil proteases responsible for cartilage matrix degradati~n as measured by the ability of _3_ neutrophil lysate, purified elastase and cathepsin G, and stimulated neutrophils to degrade cartilage matrix proteoglycan. Further, applicants have discovered that stimulated neutrophils degrada cartilage matrix in the presence of serum antiproteases indicating that degradation occurs in the serum-protected pericellular area between the neutrophils and substrate. Degradation of cartilage matrix occurring in the pericellular region c~uld be blocked only by inhibiting both elastase and cathepsin G. Applicants 1p have discovered a class of enzyme inhibitors which inhibit both elastase and cathepsin G and are thus useful in preventing neutrophil mediated connective: tissue degradation.
SUMMARY OF' THE INVENTION
Compounds of the formula P4 ~ P3 ... P2 ~. P1 _- AIM
~ ~
6 $
P4' -P3' --P2~ -P1~ -CCiTIVI
(SEQ ID NO: 1) wherein Pg is Ala, bAla, Leu, Ile, Val, Nva, bVal, Met, Nle, 30 Gly, or Sar;
P1 ~..... ~~ Ala, bAla v L~eLI r Ile, Val, NVd ~ bVal, Met, Nle, Phe, Tyr, Tyr(Me): Ala(3pyr)P Ala(4Pyr). Trp, or Nal(1):

2lU~a~~j WO 92/20357 ~ ~ ~ PCf/US92/03288 P2 is Pro, Ind, Ala, bAla, Leu, Ile, Val, N~a, bVal, Met, Nle, Phe, Tyr, Tyr(Me), Ala(3pyr), Ala(4pyr), Trp, or Nal(1);

PZ' is Pro, Ind, Ala, bAla, Leu, Ile, Val, Nva, bVal, Met, Nle, Gly, Sar or is absent;

P3 7.S LyS, Arg, Pro, Ind, Ala, bAla, Leu, I12, V81, Nva, bVal, Met, or Nle;

P3' is Ala, bAla, Leu, Ile, Vat, Nva, bVal, Met, Nle, Gly, Sar or is absent;

1~ P~ iS Alav bAla, LeU, Ile, V11, NVa, bVal, Metr Nle, or is absent;

Pg" 1S Alai bAla~ LeLI, ~l.er 'Vdl, 'IdVaobVal,, MSt, Nle, Gly, Sara or is absent;

L i s a group of one of the formulae -C~(~)-(CH2)~-C(O)--~C(O)-(~H~)~-(CH=CH)~-(C;H2)Q-C(O) - I,w--- Ph --- ~.2 -'1;1 "~Phl-~ -Ph2--L2-~C~j2)n+ 2-....therein n is 0 or an integer of from 1 to 6;

Wta 92/20357 PC,'f/U592/03288 _S_ p and q are each independently an integer of from 1 to 6;
r is 1 or 2;
L1 and La are each independently selected from a carbonyl or sulfonyl group wherein Ll is bound to the elastase inhibiting fragment and Lx is bound to the cathepsin G inhibiting fragment;
Ph, Phl, and Phi are each independently a m-phenylene or p-phenylene gr~up B is a bond, ~(C~i~)m-n or a °S(~)ZN(H)C(O)' 9rtaup;
EIM and CGIM are each independewtly,selected from the gxoup consisting of =C(O)C(O)R, _,~F~CF3,' -CF3. -CF~~', C02Ft3, -CONHR~, -CFZCHR~C(O)NHR, =H, alkyl, aryl, aralkyl, -C(O)Ft, whexein R3 is H, alkyl, phenyl; benzyl;' R is OH ar'alkAxy or a pharmaceutically acceptable salt thexenf are useful in the prevention of 'cartilage degradation:
DETAILED I~ESCRIPTTON OF THE INVENTION
Iso~teres of the compounds of f~rmula I include thbse wherein (a) one or more of the n-amino acid residues of the' Rl suhstituent is in its unnatural configuration (when there is a natural configuration) or (b) when the normal ~aeptidic amide linkage is modified,'such as for example, to form °CHaNH- (educed), -COCHZ- (keto), °CH(t~H)CHZ- (hydroxy), --C~(NH2)C~'2- (amino), -CHZCHZ- (hydrocarbon): Preferabl~r a compound of the invention should not be in an isosteric foam; p~~~icu7:arly it is px~ferred that thefe be no modified peptidic amid." gr~up in the Rl group, but if there is; it is preferable ~b keep the isosteric modification to a minimum:

2~~3~~~~
w~ ~Z~xo3s~ P(.'f/US92/03288 Unless otherwise stated, the a-amino acid building blocks of these pept~.dase substrate analogs are preferably in their L-configuration. As is conventional nomenclature used by peptide chemists, the code for an amino acid wherein the first (or other) letter of the code is upper case indicates that the amino acid has the natural "L"
configuration and wherein the first (or other) letter of the code is lower case indicates that the amino acid has '°D" configuration: Throughout this specification reference will be made to lower case amino acrd codes or codes proceeded by "(D)-" and these shall bbth be taken as equivalent.
Those compounds of this invention having aspartic or glutamic acid moieties may be in free form or a salt form, e.g., acid addition o~ anionic salt. Such a compound may be converted into its salt or base form in an art-known manner, one Erom another. preferred salts are trifluoroacetate, hydrochloride; sodium, potassium, ar 0 amm~nium salts, although the scs~pe o~ salts embraced herein is not limited thereto, the scope being extended to include all of the salts known to be used in the art of x~eptide chemistry.
~5 Before further defining and/or illustrating the scope of the peptidase inhibitors embraced by formula T; it'ma~t be convenient to state some of the more basic concepts related to peptides. Fach a-amino acid has a characteristic "R-group", the R-group being the side chain, or residue, 30 attached to the n-Garb~n atom of the a-amino acid. For exampl~'e'the R~group side chain for glydine is hydrogen;
for alanine it is methyl, for valine it. is isopropyl:
(Thus, throughout this specification the R2 mt~iety is the R-group for each indicated a-amino acid). For the specific 35 R-groups - or side chains - of the o~-amino acids reference . . -.. ..... ~'~. 1I V~7liI VJitfO
to "Principles of Biochemistry" by A.L. Lehninger, 1 S' Ed., 1982, Worth Publishers, Inc., New York is helpful.
Those compounds of formula I wherein EIM or CGIM are a -C(O)C(O)R group can exist in a hydrated or unhydrated form. Eydrates of the triketo compounds having structure I' are much more chemically stable than are the unhydrated O O I' R~ NH CH \\~ \ R
~ HO OH
R~
triketo compounds of formula I wherein EIM and/or CGIM is a -C(O)C(O)R group. For this reason, the hydrates are preferred and any reference in this specification and claims to a triketo compound should be taken to include reference to the corresponding hydrated form as context allows. Moreover, the compounds of this invention are expected to be in the hydrated form under normal physiological conditions.
The recognized abbreviations for the a-amino acids are set forth in Table I
TABLE I

AMINO ACID SYMBOL

Alanine Ala Arginine Arg Aspargine Asn Aspartic acid Asp Asn + Asp Asx ~~.U~~.~~a WO 92!20357 P(; T/US92/03288 T.~BLE I
HMINO l~CID SYMB~L
Cystein~ Cys Glutamine Gln Glutamic acid Glu Gln + Glu Glx Glycine Gly Histidine His Isoleucine I~.e Leucine Leu Lysine Lys Methionine Met Phenylalanine Pk~e p-Guanidinophenyl~lanine Phe(Gua) Proline - pro Serine Ser Thrennine Thr Tryptophan Trp Tyrosine TYr Valine Val ~orvaline Nva Norleucine Nle a .
1-Naphthylalanine Nal(1) 2-Indolinecarboxylic acid Ind Sarcosine Sar 3o Cyclohexylalanine Cha beta-Alanine bAla beta-valine bval O-4'-Methyltyrosine Tyr(Me) W~ 92/20357 PCT/U~92/03288 TABLE I

AMIN~J ACID aYM7BOL

3-Pyrazolylalanine Ala(3pyr)

4-Pyrimidinylalanine Ala(4pyr) Ns--(2-carboxybenzoyl)lysine Lys(2CBz) ~erephtholyl tP~it N~-acetyllysine Lys(AG) Applicants~prefer those compounds of f~rmula l wherein P1 is norvaline or v~line. Applicants also prefer those compounds of formula l wherein Pl is norvaline or valine;
Pl' is phenylalanine; P2 is proline; P~° is proli.ne; P3 is isoleucine, valine; or alanine; F~3' is ~lanine; valine, or is absent; P4 is a~.anine or is absent; and wherein p4' is al~hine or is absent. Applicants prefex those compounds wherein L is a -C(O)-phenylene~C(O)- group~. especially wherein the phenyl~ne is a para~phenylene group.
Applicants prefer those coanpounds of formula l wherein CGIM
and EIM is a -CF3 or -CF2CF~ group. Applicants especially prefer those compounds of formula 7: wherein ~-P,~-p~-P2--P1-(SEQ ID NO: 2) is -Ala-Ala°Pro-Val- (SEQ TD NO: 3);
LyS(2CBz)wPro-Val-; or -Val-Pro°-Val- group. Applicants , also especially prefer those compounds of formula 1 wherein -p4 ° -p3' -P~' -P~, ° - ( SEQ ID NO: 4 )' is -Ala--Ala-PrO-phe° f sES?
ID NO: 5); -Val-Pro-Phe-; or -Phe-. The most preferred compound of this invention includes WO 92/20357 ~ PCT/~.1S92/03288 C(~) _ Val ~ Pro ~ V~l _.. CF~CFg [v ~~
C~E~e~~~~s I
C(~) _ Val ..~ PrQ :r Phe ,_. CFg The peptidase'substrates of formula (T) are used'for preventing connective tissue degradation Such as cartilage degradation associated writh neutrophil associated inflammatory disease~and thus have an anti-inflammatory effect useful in the treatment of gout, rheumatoid arthritis and other inflammatory diabas~:s, and to 'prevent elastin mediated tiss~,e damage axed thus can be us~a in the area went of emphysema and adult respirat~ry disease syndrome ~a~RnS). In their end-use ap~la.cation the e~azyme 2a inhibit~ry properties of the com~AUnds of (I) are readily ascertained by standard biochemical techniques well known in'~he art> Potentia~:~d~se range fortheir and-use application will of course depend upon the nature'~nd save-rity of the disease state as determia~ed by the attending ~5 diagnostician with the range of 0:~1 to In mg~kg body, weight per day being useful for the aforeznenti~ned disease tates with 0.1 mg ~0 10 mg/kg Per day being Preferred:
xa~ing defined ~e scope of compounds embraced within ;
~ the generic formula T, the znann~r in which such c~mpounds v may be_pr.epared will;her~in below be described as illustrated: The,preparation of the compounds of formula I
may be achieved using standard chemical reactions analogously known to be useful for the preparation of a ~5 variety of known peptides. The preferred'manner o~

_ _ _.. 1 ~. 1I VJ7iI VJirOO
preparing the compounds of this invention is to first prepare fragments corresponding to the elastase inhibiting peptide of formula 2 and the cathepsin G inhibiting peptide of formula 3, P4-P3-PZ-P1-EIM c sso is xo: ~ ) 2 P4'-P~'-P2'-P1'-CGIM cssQ za xo: a) 3 l0 wherein P1, P1', PZ, P=', P3, p3~/ p1/ pl~~ ~IM and CGIM 8re as defined for formula 1, and subsequently linking the two fragments with the "L" group. The elastase inhibiting peptide of form 2 and the cathepsin G inhibiting peptide of form 3 are, in general, prepared by first preparing the compounds of structure 4 and structure 5.
P~.EIM 4 .
P~' -CGIM
wherein Pl, Pl' EIM and CGIM are as defined for formula 1.
or protected or activated derivatives thereof, and subsequently using standard techniques~known to those skilled in the field of peptide chemistry to add the desired amino acids. For this purpose, a handy reference text for t-hese techniques is "The Practice of Peptide Synthesis" by M. eodanszky and A. Bodsnszky, 1985, John Wiley and Sons, lnc. New York, wherein the parameters and 3 0 techniques affecting the selection, use and removal of protective groups for individual and groups of amino acids is detailed, and which also contains activation and coup;ing techniques and other special procedures. However, before the application of these peptide chemistry techniques may be applied, 3 5 certain key intermediates containing the elastase inhibiting moiety WO 92/20357 ' ~ . . ' PG'f/IJS92/0328Fs -lz-(EIM) and the cathepsin G inhibiting moiety (CGIM) must first be prepared. The preparation of the key intermediates is described as follows.
Those compounds of formulae 2 or 3 may be prepared using standard chemical reactions analogously known in the art. More specifically the compounds of formulae 2 and 3 wherein EIM and CGIM are -CF~H, -CF3, -CU2R3, -CONHR3, -C(U)R, -CFaCHR3G(O)NHR, H, alkyl, aryl, or aralkyl are known in art. Thus a description of the preparation of the compounds of formulae 2 or 3 wherein EIM or CGIM represents -CFZH, -CF3, -CU2Rg, -CONI3R~r or -C(U)R can be found in European Patent Application Numbe~c 195,212, published September 24, 1986. A description of the preparation of 1S the compounds of formulae 2 or 3 wherein EIM or CGIM
represents -CF2GHR3C(O)NHR are described in European Application Numbex 275,101, publ~,shed July 20, 1988. A
description of the preparation oi_ the'compounds of f~rmulae 2 and 3 wherein EIM and C~IM represents H, alkyl, aryl, or aralkyl are described in European Patent Application Number 363,284, published April 11; 1990.
The preparation of the compounds of formula 2 wherein EIM or CGIM represents -C(0)C(0)R a.s outlined in Scheme A
wherein R, P~,, PZr P3; and P4 are as previously aef~.ned.
The compounds of formula 3 can be prepared ~:n an analogous manner. Specifically; the dompounds of formula 2 can be prepared by treatment of the appropriate glide of f~rmula 6 with (a) ozone and dimethyl sulfide or (b) ringlet oxygen.
The ozonylysis readtion ca.n be conveniently performed by, for example, bubbling an excess of ozone through a cooled solution of the appropriate formula 6 glide. Su~.table solvents include any r~onreactive solvent in whidh the formula 6 glide is soluble, for example, alkyl esters of simple alkanoic acids such as ethyl acetate; the WO 92/20357 PGT/US92/032~~

chlorinated hydrocarbons such as carbon tetrachloride, chloroform, 1,2-dichloroethane, 1,1,2,2-tetrachloroethane, and methylene chloride; the aromatic hydrocarbons such as benzene, toluene, and xylene; a chlorinated aromatic such as 1,2,4-trichlorobenzene and o-dichlorobenzene; an alcohol such as methanol, ethanol, and isopropanol; or an ethereal solvent such as diethyl ether, tetrahydrofuran (THF), and 1,4-dioxane. Methylene chloride is preferred.

WO 92/20357 P~/U592/032~$
E$EaCtl011 ~Ch~(Yie /~
X
PhtN - P1 - OH I
CH2~ R 10 O v 1) ~3P
CIzCHOCH3 a 2) base $3P
PhtN ~. P 1 ~. CI R
I!
P~3 PhtN ~ P 1 R
'!) excess H2NNH2 2) P~, P3, Peg c~u~ale p~3 P4_-P3~.,P2~P~ R 6 t 4 ~~
~ (SEQ,ID N~O: 9) 1) 0~ . or Sin~let Oxygen 2) (CH3)zs ~O ~~
P4 "~. P3 .a P~ ,s,. P1 R 2 (EI~=C(OH)2COR) (SEQ ID NO: 10) ~~~.~~ ~.~~.FJ
V1~0 92/20357 PCTlUS92/03288 The temperature of the ozonolysis reaction mixture can be any temperature conducive to the reaction, typically from about -78°C to about 0°C, preferably from about -78°C
to about -35°C, and most preferably about -70°C. The time of the reaction will vary depending on the glide, the concentration of the reactants, the temperature and other factors. Conveniently, ozone is bubbled into the reaction mixture until the solution turns blue indicating an excess of ozone.
The ozonide is theh treated with an excess.~f a reducing agent such as zinc metal or preferably dimethylsulfide. The desired formula, 2 compound as the hydrate is isolated from the reaction mixture in-any convenient manner, typically by solvent removal (via evaporation). Purificat~.on may be accomplished by, for example, flash chromatography.
Oxidati~ns uta:lizing ringlet 'oxygen are well known.
More specificallyr ringlet oxygen oxidation ~f an glide to produce a tricarbonyl ester has been reported by H:
Wasserman et al.: J. Amer: Chem, 8oc. 0'1, 371 (1989).
Singlet oxygen can be generated by dye-sensitized excitation of oxygen. Suitable dyes include Rose Bengal, Eosin Y and methylene brae: Other sen~itizers include dinaphthalenethiophene. Typically, Rose Bengal and Eosin Y
are attached to a basic anion-exchange resin and methylene blue is attached to an acidic ration-exchange resin.
3p Excitation is accomplished with a UV lamp such as a tungsten,-iodine lamp. Suitable solvents are any solvents which promote and do not interfere with the desired reaction. Such eolver~ts a.nclude the aromatic hydrocarbons such as benzene and toluene; hydrocarbons such as hexane;
85 ethereal solvents such as diethyl ether, tetrahydrofuran ~ :~. 4~ ~ a ~ ~~ ~ . .
wo yx~xo3s7 ~C,'C/US9x/0328b (THF) 1,9-dioxane; chlorinated hydrocarbons such as dichloromethane and chloroform; carbon disulfide; and alcohols such as methanol, ethanol, propanol, isopropanol and t-butanol. P~lixtures are operable. The temperature of the reaction mixture can be any suitable temperature from about -?~°C to about 30°C typically from about -78°C to about -50°C. The time of the reaction will vary depending on the reactant, the solvent, concentrations, and temperature and can be from about 1 min to about 2 hours.
purification and isolation can be by those methods described above for specification and isolation of product from the ozonolysis reaction mixture.
The formula 6 glide is prepared from the appropriate N-protected glide, preferably from the phthaloyl protected glide of formula ?. The removal of the phthaloyl group can be readily achieved by methods generally known to those skilled in the art. For example~ a solution of the phthaloylglide can be allowed to react with hydrazine hydrate, typically about a 20-fold excess of hydrazine hydrate, until the reaction is substantially complete. The solvent can be any of those described above for the ozonolysis reaction and preferably will be an alcohol solvent such as Et~H: The emp~rature of the reaction mixture can be from about 0°C to about 60°C, conveniently at about room temperatureo i.e., 25°C: The reaction time will vary depending on the specific reactant, the temperature, the solvent, and other factors known to influence reaction dime. Conveniently, the pogress of the 3p reaction can be monitored by thin layer chromatography ( TLC ) o ,~..
Subsequent to removal of the phthaloyl group, the P2, P3, and P4 groups can be linked to the now free amino group.

~~.a~<~~_' WO 92/20357 PC.°f/LJS92/03288 The PZ, P3, and P4 can be linked to the unprotected, free amino compound by well known peptide coupling techniques.
In coupling individual amino acids or peptides to the deprotected formula ? compound, appropriate side chain protecting groups are employed. The selection and use of an appropriate protecting group for these side chain functionalities is within the ability of those skilled in the art and will depend upon the amino acid to be protected and the presence of other protected amino acid residues in the peptide. The selection of such a side chain protesting group is critical-in that it must not be remoeed during the deprotectior~ and coupling steps of the synthesis. For example, when Boc is used as the ce-amino protecting group, the following side chain protecting groups are suitablea p-toluenesulfonyl (tosyl) moieties can be used to protect the amino side chains of amino acids such as Lys and ~.rg; r~--methylben~yl, acetamidomethyl, benzyl (Bzl), or t-butylsulfonyl moieties can be used to protect the sulfide containing side chains of amino acids such as cysteine, h:omocysteine, penicillamine and the like pr derivati.ves thereof; benzyl (B~1) or cyclohexyl ester moieties can be used to protect carboxylic acid s~.de chains of amino acids such as lisp or Glu; a benzyl (Bzl) ether can be used to protect the hydroxy containing side chains of amino acids such as Ser and Thr; and a 2-bromocarbobenzoxy (z(Br)) moiety can be used to protect the hydroxy containing side chains of amino acids such as Tyr. These side chain protecting groups are added and removed according to standard practices and procedures well known in the art. .
It is preferred to depr~tect these side chain protecting groups with a solution of anisole in anhydrous hydrogen fluoride (1:10). Typically, depratecti~n of side chain protecting groups is performed after the peptide chain synthesis is complete but these groups can alternatively be 2~~J~~~.
'~U~ 92120357 ' PC'f/US92/03288 _18_ removed at any other appropriate time. It is preferred to deprotect these side chains at the same time as the peptide is cleaved from the resin when solid phase synthetic methods are employed.
The phthaloyl glide Qf formula 7 is prepared by reaction of the phthaloyl protected acid chloride of formula 8 with the phosphonium glide of formula 9. This reaction is performed by adding a solution of the appropriate formula 9 glide, preferably dropwise. to a solution of the formula 8 acid chloride. Suitable solvents include.those listed above for-,the ozonolysis reaction and will preferably be an ethereal solvent such as THF. The reaction will require fro~zn abouf 30 minutes to about l2 hours, typically about 2 to 3 hours, depending on the acid chloride, the glide, the solvent(s~, and the temperature which can be from about 0°C to about 60°C, conveniently at about room temperature, i.a., 25°~C. Is~lation and purification is accomplished by filtering the-reaction mixture to remove solid products and subsequently chromatogr;apning the fi:ltr~t~, for example; on silica gel eluting with a 50~ mixture of ethyl acetate and hexane.
The formula 9 phosphor~aus glide; Wittig reagent, is prepared from the corresponding formula 10 a°halocarboxylic acid derivative in the usual manner, that is; by reacting the a-halo ester with a tertiary phosphine such as triphenylghosphine to yield a phosphonium salt. When treated with a str~ng base such as an organoli.thium compound, for example, lithium diisopropylamide (LDA), s~dium hydride, or sodium amide, the acidic proton is removed"..end the desired ylide'is formed; Suitable s~lvents used in forming the Wittig reagent include any nonreactive solvent, for example, the aromatic hydrocarbons such as benzene or toluene, the ehlorinated hydrocarbons such as of ~ ~ a VI~O 9Z/20357 PC.°f/~LJS92I03288 carbon tetrachloride, chloroform, or methylene chloride, or the ethereal solvents such as diethyl ether or THF.
The reaction can conveniently be performed at from about 0°C to about 60°C, typically at room temperature, that is about 25°C. The halo group of the a-halo ester is preferably a bromo group, but can be a chloro or iodo group or can be any good leaving group which forms a stable phosphonium salt such as a mesylate or tosylate group.
to The acid chloride of formula 8 is prepared from the corresponding acid of formula 1l by, for example. reacting the acid with .refluxing a,a~-dichloromethyl methylether.w After about 3 hours, the solution is~allowed to GOO1 and the product concentrated by solvent evaporation. The resulting crude acid chloride can be used directly without further purification in the reactian with the formula 9 phosphorous ylide.
The preparation of the compounds of formula 2 wherein EIM or CGIM represents -CF~CF3 is outlined in ScYaem~ 8 wherein R~ and R2 are as previously defined, and Pg i~ an amino protesting group such as a carbamate, preferably a benzyloxycarbonyl (Cbz) group.

WO 92/20357 1P(.T/US92/032g~
Reaction Scheme R
p ~ P1 ~ ~ o OCH3 ~. OCH3 g P4~P3wpz~pl ~~
95a ' 4 ~ 1~b ( S~t~ ID No: 11 ) CH3 I.AHlTHF 'LAHITHF
Pg~P9 -~.H , P~a~3 ~P2_p1 ~H
14a 14b (SEQ ID tdo: 12) LiCF~CF3/Ether i-iCF2CF31Ether ~
I:iCF2CF~lEther ~9NH
~' CF2CF3 P,~_2 -a \ CFzCF3 13a R2~C 1) Ci~avage ~f p9 RZSC 13b 2) Co~rpling ofi PZ~~
Oxidation (~~a ~m ~o: u3~
pxidation ... . 1) Cleavage of lag pg _ P~ ~CFzCF~ .~.-r--:~.-,~ . 2 ~~t~t1=CFzCF~,) 2) Cpupiing ofi P2_4 2 ~.. '~ ~ ~. ~~
WO 92/20357 PC,T/1JS92/03288 The compounds of formula 3 can be prepared in an analogous manner.
Specifically the compounds of this invention are prepared by reducing the N-methoxy-N-methyl amide of: either formula 15a or 15b to produce the aldehydes of formulae 14a and 14b, respectively. Applicants prefer to use the compounds of formula 15a as the initial starting materials.
The reduction can be performed in any way generally known la and readily performed by those skilled in the art such as by use of lithium aluminum hydride (L,~H). This reduction can bevconveniently carried out by adding aw excess.~f LAFI
to a cooled~ typically about 0°C, solution. of a foranula 15a or 15b compound in a nonreactive solvent such as an ethereal solvent such as tetrahydrofuran (THF). P.fter the reaction is substantially complete, typically after about 30 minutes. the reaction mixture is quenched by the addition of, for example, 10~ potassium hydr~gen sulfate and then water. The product can then be isolated by, for 2p example, extraction of the aqueous mixture with a solvent such as ethyl acetates drying and solvent removal. The crude product can be purified by, for exampleo column chromatography such as a silica gel column eluting with 55$
ethyl acetate/hexane or recrystallization.
The formulae 14a and 14b aldehydes are then reacted d with the pentafluoroethyl anion, such as the lithium salt of the pentafluor~ethyl anion to gisre the alcohols of formulae 13a or 13b, respectively. This condensation can 3p be conveniently prefarmed by those skilled in the art by a.
modified procedure as described by Po G. Gassmaz~ and Neil J. O'Reilly, J. Org. Chem. 1987, 52y 2481-2490. zn thus procedure, the p~rfluoroethyl anion is generated irasatu by addition of methyllithium/l.ithi:um bromide complex to a 35 solution of the aldehyde and pentafluoroethyl iodide in a WHO 92/20357 PC'f/1JS92/0328~

nonreactive solvent such as diethyl ether. The cooled (-78° - 0°C) reaction mixture is allowed to stir for about one-half to about l hour or until the reaction is substantially complete and then the mixture is quenched by pouring into an excess of dilute hydrochloric acid. The product is isolated by, for example, extraction with diethyl ether and subsequent solvent removal. The crude product is purified by, for example, chromatography on silica gel.
The alcohols of formulae 13a or 13b are then oxidized to give the amino°protected pentafluoroethyl keton~s of formula 12 or the desired product of,formula 2, ,.
respectively. The oxidation may be effected via the well-known Swern oxidation procedure, or with a modified Jones oxidation using pyridinium dichromate, or a chromic anhydride-pyridinium complex, or with the Dess-Martin periodinane, 1,1,1-Iris(acetyl~xy)-l,l--dihydro-1,2°
benzoiodaxol-3(1H)-one. ~f cburse, if there are any ZO protecting groups on the residues of the q-amino acid building blocks, such protecting groups may be removed after oxidation. The coupling,procedures are effected according to standard pr~ced~re~ well known a.n the art.
In general the Swern oxidation is effected by reacting about 2 to 10 equivalents of dimethylsulfoxide (DMSO) with about 1 to 6 equivalents of trifluoroacetic anhydride [(CF3C0)a0] or oxalyl ch~.oride [(COC1)2], said reactants being dissolved in an inert solvent, e.g.. methylene chlo-ride (CH2C1~), said reaction being under an inert atmosphere (e. g., nitrogen or equivalently functioning gas) under anhydrous conditions at temperatures of abou -80°C to --50°C to form an in situ sulfonium adduct to which is added about 1 equivalent of ain appropriate alcohol of formula 13a or 13b. Preferably, the alcohols ire dissolved in an .inert solvent, e.g.. CH'ZCl= or minimum amounts of DMSO, and the reaction mixture is allowed to warm to about -50°C (for about 10-20 minutes) and then the reaction is completed by adding about 3 to 10 equivalents of a tertiary amine, e.g..
triethylamine, N-methylmorpholine, etc.
In general-, the modified Jones oxidation procedure may conveniently be effected by reacting an alcohol of formula 13a or 13b with pyridinium dichromate by contacting the reactants together in a water-trapping molecular sieve powder, (e. g., a powdered 3 Angstr8m molecular sieve), wherein said contact is in the presence of glacial acetic acid at about 0°C to 50°C, preferably at room temperature followed by isolation and then optionally removing amine protecting groups.
Alternatively, 1 to 5 equivalents of a chromic anhydride-pyridine complex.(i.e.. a Sarett reagent prepared insitu (see Fieser and Fieser "Reagents for Organic .
Synthesis" Vol. 1, pp. 145, 1976, John Wiley and Sons, Inc. New York and Sarett, et al., J.A.C.S. 25, 422, (1953)) said complex being prepared in situ in an inert solvent (e.g., CH2C12) in an inert atmosphere under anhydrous conditions at 0°C
to 50°C to which complex is added 1 equivalent of an alcohol of formula 13a and .
13b allowing the reactants to interact for about 1 to 15 hours, followed by isolation and optionally removing amine protecting groups.
Another alternative process for converting an alcohol of formula 13a or 13b to the desired ketone of formula 1 or

5 is an oxidation reaction which employs Dess-Martin periodinane (see Dess and Martin, J. Or . Chem.. 48, 4155, (1983)). This oxidation is effected by contacting about 1 equivalent of the appropriate alcohol of formula 13a or 13b with 1 to 5 equivalents of periodinane (preferably 1.5 equivalents), said reagent being in suspension in an inErt ~~.~~a~.~
'1y0 92/20357 P~If'/US92J03288' solvent (e. g., methylene chloride) under an inert atmosphere (preferably nitrogen) under anhydrous conditions at 0°C to 50°C (preferably room temperature) and allowing the reactants to interact for about l to 48 hours.
Optional deprotection of the amine protecting groups may be effected as desired after the ketones have )assn isolated.
In one mode of preparing the compounds of this invention the formula c~mpounds are prepared by first converting the amino-protected, perfluoroethyl alcohol of formula 13a to the corresponding compound of formula 13b, grior to ffinal oxidation The amino-protected, perfluoroethyl alcoh~1 of formula 13a is ffrst c~eprotected, if desired, and then any amino'acids or peptide chain represented by P4-p3-PZ- can be added using standard a°amino acid or peptide coupling procedures. Where the ~q-F3-P2--group is made up of more than one amino acid, either the entire peptide chain can be added to the deprotected formula 13a compound'or the amino acids can be coupled ~o the deer~tected formula 13a compound sequentially.
Alternatively, a combination of these two coupling methods can be used. In a like manner; the compounds of formula 12 can be converted to the desired formula 2 compounds.
In coupling individual amino'acids or peptides to the deprotected f~rmula 13a or formula 12 compounds, appropriate side chain protecting groups are employed. The selection and use of an appropriate protecting group for these side chain funct'ionalities is within th,e ability of 3p those skilled in the art and will depend upon the amino acid to..be protected and the presence of other protested amino.acid residues in the peptide. The selection of such a side chain protecting gx~~ap is critida7. in that it must not be removed during the deprotectian and coupling steps of ~;he synth.esis. ~°or example, when Hoc is used as the a-~~..~:~ a~~
WO 92/20357 PO'flUS92f032~~

amino protecting group, the following side chain protecting groups are suitable: p-toluenesulfonyl (tosyl) moieties can be used to protect the amine side chains of amino acids such as Lys and Arg; p-methylbenzyl, acetamidomethyl, benzyl (8z1), or t-butylsulfonyl moieties can be used to protect the sulfide containing side chains of amino acids such as cysteine, homocysteine, penicillamine and the like or derivatives thereof; benzyl (~zl) or cyclohexyl ester moieties can be used to protect carboxylic acid side chains of amino acids such as Asp, flu; a benzyl (~zl) ethex can be used to protect the hydroxy containing side chains of amino~acids~
sueh as Ser and Thr;.and a 2-bromocarbobenzoxy (ZBr-Z) moiety can be used to protect the hydroxy containing side chains of amino acids such as Tyr. These side chain protecting groups are added a~ad removed according to standard practices and procedurias well known in the art.
It is preferred to deprotect ~h~se side chain protecting gfoups with a solution of anis~le in anhydrous hydrogen fluoride (1:10). Typically, deprotection of side chain protecting groups is performed after the peptide chain synthesis is complete but these groups can alternatively be removed at any other appropriate time. It is preferred to deprotec~t these side chains at the same time as the peptide is cleaved from the resin when solid phase synthetic methods are employed.
In the preferred mode o~ preparing the compounds of this invention, the compounds of formulae 15a and 15b can be converted directly to the compounds of formulae 12 or 2;
respect~..v.ely, by condensation of the N-methoxy-N-methyl amide with the lithium salt of the perfluoroethyl anion in the same manner in which the compounds of formulae 15a ~r~d 15b are converted to the compQUnds of formulae 14a and 14b, respectively.

Vd~ 92/20357 ' PC'T/US92/~3288 The compounds are then isolated and purified by standard techniques. The desired amino acids, derivatives and isomers thereof can be obtained commercially or can be synthesized according to standard practices and procedures well known in the art.
The N-methoxy-N-methyl amides of formulae 15a and 15b are prepared fr~m the corresponding n-amino acids of formulae 16a and 16b, wherein Rl and R2 are as defined for formula 1 and wherein Pg is an amino protesting group such as carbamate, preferably a benzyloxycarb~nyl (Cbz)' group.
respectively, in the usual manner. (See, for example; J.A.
F'ehrentz and B. Castro, S~nthes~.s, 676-78 (1983).
p9NH ~ ~ OCH3 R~NH / ~CH3 N
R2 CH3 Rz CH3 9 6a 16b zsobutylchloroformate is added to a cooled (i.e. -60-gC-to about 0°C) mixture'of N~methylmorpholin~ or another sterically hindered, non-nucleophilic tert~.ary amine and an a-amino arid compound in a nonr~ctzve solvent such as methylene chl~ride: After about 5 minutes to about 1 hour, typically about 15 - 2O minutes, N,O-dimethylhy~droxylamine Fi~l is added and the mixture allowed to stir for from ab~ut 30 minutes up ~o about 6 hours and then the reac:t.~on wo ~zizo~s7 Pccivs~zro~zs~
_2~_ 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 phase is extracted with, for example, ethyl acetate. The desired compound is then isolated by solvent evaporation and crude purification can be accomplished by, for example, flash chromatography on silica gel eluting with ethyl acetate/hexane.. Purification can be acomplished by, for example, flash chromatography on silica gel eluting with methylene chloride.
The compounds of formula (1) wherein Ll and L~ are bbth represented by carbonyl groups, Ll and L2 are both represented by sulfonyl groups or L2 is represented by a carbonyl group and Ll is represented by a sulfonyl group can be prepared by techniques and procedures well known an appreciated by one of ordinary skill in the art. A'genersl synthetic scheme for preparing t:he~e compounds of formula (1) is set forth in Scheme ~C. ~n Scheme G, all substituents unless otherwise iudidat~d are as previoulsy defined.

2~(~~~~~
WO 92/20357 ~~'/US92/0328F~ :.:;~' Scheme C
COUPLING
4 L" (17) 1 4 Pa-P~-PZ-P1-EIM ~ L'-Pa-Pg-P2-P1-EIM
( sEQ ID No: ~ ) (~) step a ( sEQ ID ~o: is ) (1$) COUPLING
I a-p3vP2--P1°EIM
Pa ~ P3 ~ P2 ~ P1''~f~IM t ~ ) , L

( SEQ ID ~to: 8 ) Step b ! _~ -P ~P _CGII~I
a° 3~ 2' 1' (gEQ ID N0: 1) ~1) L" = an appropriate di-functionalized derivative of L
L' = an appropriate mono-functionalized derivative'of L
Scheme C provides a general synthetic;procedure for preparing the compounds of formula (1) wherein L1 and L2:are both represented by carbonyl groups; L~ and L2 are both represented by sulfonyl groups or ~z is represented by a 30 carbonyl group and L~, is represented by a sulfonyl gr~up.
In step a, the appropriate elastase inhibiting peptide fragment of .formula (2) is coupled with the appropriate . derivative of L as described by structure (17) to give the ~v~~ 3~_~~
W~ 92/20357 PCT/US92/03288 corresponding L-elastase inhibiting peptide fragment of structure (1g) by techniques well known in the art.
When the compound of formula (1) is one wherein Ll and s L2 are both represented by carbonyl groups, an appropriate derivative of L as described by structure (17) is one wherein the LZ carbonyl group is represented by a t-butyloxycarbonyl prote~ted.carboxylic acid and the L1 carbonyl group is represented by an unprotected carboxylic acid.
When the compound of formula (T) is one wh~rain L1 and LZ are both represented by sulfonyl groups, an appropriate derivative of L as described by structure (17)'is one wherein the L1 sulfonyl group is represented by a sulfonyl chloride group and the LZ sulfonyl ~r~up is represented by an unprotected sulfonic-aCia.
When the compound of formula (1) is one wherein L2 is represented by a 'earbonyl group and L1 is represented by a sulfonyl group, an appropriate derivative of L as described by,structure (17) is one wherein the L2 carbonyl group is represented by a t-butyloxyca~bon'y1 protected carboxylic acid and the L1 sulfonyl'group is represented by a sui;fonyl ch~:oride groupe rn step b, the appropriate L-elas~as~ inhibiting PePticle fragment of structure (1g) is coupled with the appropriate cathepsin G inhibiting peptide fragment of formula (3) to give the corresponding compound of formula ( 1 ) by techniques well 'kn~~rn in the art .
When the appropriate L-elastase inhibiting peptide fragment of structure (1g) is one wherein the L~ carbonyl group is represented by a t-butyloxycarbonyl pr~t~ected ~~1~~~~
WO 92/20357 ~ ' PCT/U592/0328L
. ~ -30-carboxylic acid, the t-butyloxycarbonyl protected carboxylic acid must first be hydrolyzed by techniques well known in the art prior to the coupling reaction in step b.
Starting materials f~r use in Scheme C are readily available to one of ordinary skill in the art.
The compounds of formula (1) wherein L1 is represented by a carbonyl group and L2 is represented by a sulfonyl group can be prepared by techniques and procedures well known an appreciated by one of ordinary skill in the art.
A general synthetic scheme for preg~aring here c~mpounds of formula (1) is set forth in Scheme D:, In Scheme D, all substituents unless otherwise indicated are as previously defined.

WO 12/20357 ~ ~- ~ ~ ~ ~- '~ PC.'fI~.JS9z/0328~

Scheme D
COUPLING
Pq'-P3'~-P2°-P1'--~GI1H Lae (19) L,-Pro-P3~_p~°_pI,_~Glii ( sE~ In r~o: s ) ~3~ step a ( s~Q Ian ~o: is > (20) COUPLING
Pa-~3=PZ-PI-EIM
P'~ p3~p2 PZ-EIM (2) Lo SEQ ID td0: 7 ) Step b ~q a ~p3 i _p~ ~ ~Pl o _CGIM
(SEQ ID' NO~ 1) ~1~
L°' = an,appropriate di-functionaliaed derivative of L.
L' - an appropriate m~no-fux~ctionali~ad derivative of L
Scheme D pra~ides a general. synthetic procedure for preparing the compounds of formula ( 1 ) ~lh~rei.n L1 is represented by a carbonyl group anc~ Lz is represented-by a sulfonyl group In s~.ep a, the appropriate bathepsin G ~:nh~.biting-peptide fragment of formula (3) is coupled with ,the appropriate derivative of'L as described by structure:(19) to give the corresponding cathepsin G inhibiting peptide WO 92/20357 PCT/US92/032~~

fragment of structure (20) by techniques well known in the art.
An appropriate derivative of L as described by structure (I9) is one wherein the Ll carbonyl group is represented by a 't-butyloxycarbonyl protected carboxylic acid and the LZ carbonyl group is represented by a sulfonyl chloride group.
In step b, the appropriate cathepsin G inhibiting peptide inhibiting peptide fragment of structure (20) is coupled with the appropriate elastase inhibiting peptide fragment of formula (2) to give the corresponding compound of formula (1) by techniques well known in the art.

The t-butyloxycarbonyl protected carboxylic acid functionality on Ll of the appropriate cathepsin G
inhibiting peptide inhibiting peptide fragment of structure (20) must first be hydrolyzed by techniques well kn~wn in the art prior to the coup~:ing reaction in step b.
starting materials for use'in Scheme D are readily available to one of ordinary skill in the art.
;2~ The following speca.fic examples are given t~ illustrate the preparation of this invention although the scrape of compounds is not meant to be limiting to the scope, of compounds embraced by formula I.

a WO 92/20357 ~ ~ ~ ~ v ~ ~ PCT/US92/03288 (CFA]Phe-Pro-Val-C(0)-phenylene-C(O)-Val-Pro-Val[CF2CF3]--SEQ ID NO: 6 Preparation of Hoc-Val[CF2CF3]
Dissolve Boc-Val dimethylhydroxyamide (1.0g. 3:8mmo1) in ethyl ether (50mL) and cool to -78°C. Add pentafluoroethyl iodide (3g, 12.2mmol) followed by methyllithium~lithium bromide complex (6mL of a 1.5M solution). Rbpeat the addition of pentafluoroethyl iothde (3g, 12.2mmo1) followed by methyllithium~lithium bromide complex (6mL of a 1:5M
solution) three times: Stir for l5~minufes at -78°C then allow to warm to room temperature. Pour into water and separate the organic phase. Extract the aqueous phase with ethyl ether (3X150mL), combine the organic phases and dry (Na2S'Q4 ) ~ Evaporate the ~olver~t in va~uo and purify by silhca gel chromatography (10% ethyl acetate/hexane) to give the title compound.
Preparation of VaIjCF3CF~ ~hydrochlorid~
Dissolve Boc-Val[CFZCF3] (350mgl:lmmol)' in ethyl acetate (50mL) and cool to 0°C< Treat with hydrogen chloride gas for 5 minutes and stir for 30 aninutes. Remove the solvent in vczcuo to give the title compound:
Preparation of Hcac-Val-Pro-Val[CF~CF~]
Dissolve Boy-Val-Pro (314mg; l.Ommol) in me~hylene chloride (4mL) and add N-methylmorpholine (252mg, 2:5mmol). Cool t~
-22°C and add isobu~ylchloroformate (136mg~ l.OmanoZ). Stir for 20.~minutes and add to Val(CFZCF3]~hydroch~oride (l.lmmol). Stir for 1 hour at °22°C, allow to warm to raom temperature and stir for 3 hours. Purify by silica gel chromatography (40~ ethyl acetate/hexane) to give the tile compound (405mg).

~:~0~~.~f WO 92/2035? ~ PCT/US92/03288 Preparation of Val-Pro-Val[CF2CF3)~hy~.rochloride Dissolve Boc-Val-Pro-Val[CFZCF3] (385mg. 0.?4mmo1) in ethyl acetate (50mL) and cool to 0°C. Treat with hydrogen chloride gas for 5 minutes and stir for 30 minute .
Evaporate the solvent invdcuo to give the title compound (334mg).
Pre~aaration of Boc--Val-Pro-Phe [ ~H ] [ CF3 l Mix 2-ghenyl-~2°oxazoline-4-phenylmethyl-5-one (Synthesis, #3, 191-3, (1982)) (300g) end trifluoroaGetic anhydride (?OOg). Heat at relax far 3 hours then stir overnight at room temperature. Evaporate the solvent in vccuo and add oxalic acid (400g). Stir and add additional oxalic acid (50g). Heat until evolution of C~2 ceases and a solid forms.
Cool to room temperature and dissolve in a 1:3 mixture of water,/ethyl acetate (12L). Segarate the organic phase, wash until basic with saturated sodium hyd~rog~n carbonate then with water. Dry (MgS04): filter and concentrate by boiling to a volume of 2.5L. Cool to room temperature and add hexane (1L). Filter the precipitated solid and air dry to give 1.1.1-trifluora-2-one-3-ben~oylamino--4-phenylbutane (18?.6g).
Dissolve 1.1.1-trifluoro-2-one°3-benzoylamino-4-phenylbutane (187.6g) in ethanol (1L) and cool in an ice bath. Add sodium borohydride (11g) in portions over 15 minutes. Remove the ice bath and stir at room temperature to l.5 hours. Replace the ice bath and carefully treat with 10~ hydrochloric acid (250mL). Add ethyl acetate (4L) to dissolve and then add water (500mL). Separate the organic phase, wash with brine (4X300mL)'and dry (MgS04):
Filter and evaporate the solvent invacuo. Add hexane and filter to give 1,1,1°trifluoro-3--benzoylamino°4-phenyl-2-butanol as a white solid (145.2g).

WO 92/20357 PC."T/1JS92/0328~
°35-Mix 1,1,1-trifluoro-3-ben~oylamino-4-phenyl-2-butanol (145.2g), concentrated hydrochloric acid (1.4L), water (700mL) and ethanol (1L). Heat to reflux for 24 h~urs then add additional concentrated hydrochloric acid (400mL) and ethanol (1.2L). Stir an additional 24 hours. Evaporate the ethanol in vczcuo and filter > Cool the filtrate to room temperature and treat with sodium hydrogen carbonate and then with 50~ podium hydroxide while cQOling in an ice bath. When pH IO is obtained, filter off the solid to give [CF3][OH]-Phe (5S.2g), Dissolve Boc-Val-Pro (3°3g, 10>5mmol) in methy~:en~ chloride (25mL) and add N-methylmorpholine (2.128,. 21mmo1). Cool to~
-22°C and add isobutylchloroformate (1:438, 10:5mmolj.
Stir at -22°C for 25 minutes then add [CFA][OH]-phe (2:Sc~.
11.5mmo1). Stir at -22°C f~r 3 hours, allow to warm to room temperature and stir over.ni.ght. Poux into wader (100mL) and extract into ethyl' ether (3X150mL)Wash tlae combined organic phases wi h dilute hydrochloride acid then saturated sodium hydrogen carbonate. Dry (Na2S04) and evaporate the solvent invcxcuo. Puri-fy by silica gel chromatography (40~ ethyl acetate/h~xane) to give the title compound (5.27g)>
P~~axation of Boc-Val-Pro-Phe[CFA]
Dissolve Boc-~lal-Pro-Phe[OH][CF3] (0.79g. 1:54mmo1) in methylene chl~ride (25mL) and add Dens-Martin reagent (2.5g). Stir at room temperature overnight then pour into 50mL of water c~ntaining sodium hydrogen carbonate (loOg) and sodium bisulfite (1.7g): Extract with ethyl ether ( 3X100mT:) and dry (Na~SO$ ) . Evaporate the solvent in vcxcuo and purify by silica gel chromatography (40~ ethyl a~etate/hexane) to give the fiitle compound (755ang)>

~~.~~~1 WO 92/20357 P~f/US92/0328~
-36°
Preparation of Val-Pro-Phe[CFA ~hydrochloride Dissolve Boc-Val-Pro-Phe[CFg] in ethyl acetate (100mL) and cool to 0°C. Treat with hydrogen chloride gas for 5 minutes and stir at 0°C far 30 minutes. Evaporate the solvent invacuo to give the title compound (840m8).
Preparation of Boc-phenylene-C(O)-Val-Pro-Phe[CFz]
Dissolve Boc-phenylene-C(O)OH (370m8, 1.67mmo1) in methylene chloride (4mL) and N-methylmorpholine (0.18mL, 1.67mmol). Cool to -20°C and add isobutylchloroformate (0.2278, 1.76ntmol) ans stir for 45 minutes.. Add methylene chloride (2mL) and add N-methylmorpholine (0.18mL) and Val-°
Pro-Phe[CF3 ] hydrochloride 0.758, 1.67mmol). Stir at -20°C
for 2 hours, allow to warm to room temperature and stir for an additional 3 hours. Pour into a mixture of methylene chloride (lOmL) and water (20mL). Separate the organic phase and extract the ague~us phase with methylene chloride (2X20mL). Combine the organic phases and dry (Na2SOa).
Evaporate the solvent invczcuo and purify by silica gel chromatography to give the title compound (575m8).
Fre~aration of HOC--phen~ylene-CEO)-Val-Pro-Ph~ CFA ] ~h~rdrochloride Dissolve Boc-phenylene-C(O)°Val-Pro-Phe[CF3] (250m8) in ethyl acetate (50mL) and cool to 0°C. Treat with hydrogen chloride gas for 5 minutes and stir at 0°C for l hour.
Evaporate the solvent inuacuo to give the title compound (232m8).
Pze aration of CF Phe-Pro-Val-C O - hen lane-C O -Val-Pro-Val[CF2GF~]--SEQ ID NO: 6 Dissolve B20C-phenylene-C(O)-Va1-Pro-Phe[CF3]~hydrochloride (230m8, 0.41mmol) in methylene chloride (3mL) and add N-methylmorpholine (41.4m8, 0.41mmo1). Cool to -20°C and add isobutylchloroformate (55.7m8, 0.41mmo1). Stir ft~r 45 minutes at -20°C and add Val-Pro-Val[CFZCF~]~hydrochloride (lSSmg, 0.41mmo1). Stir at -22°C for 3 hours, allow to warm to room temperature and stir for an additional 3 hours. Pour into water and extract into methylene chloride (3X25mL). Combine the organic phases and dry (Na~S04).
Evaporate the solvent ira Uaccuo and purify by silica gel chromatography (ethyl acetate then methanol) to give the title compound (190mg).
Example 2 CF3 Phe-C 0 - hen lens-C 0 -Val--Pro-Val CF CF -SE ID N~s Preparation of Boc-phen~lene-C(O)-Phe[OH]jCF~]
Dissolve Boc-phenylene-C(0)OH (0.615g, 2.8mmol) in methylene chloride (6mL) and add N-methylmarpholine (0.6g).
Cool to -22°G and add isobutylchloroformate (0.4mL, 3.O~mmol). Stir at °22°C for 25 minutes and add a ~o~.ution of Phe[OH][CF3] (0.64g~ 2.9mmol) in methylene chloride (2mL) and N-methylmorpholine ;0.3g). Stir at -22°C for 1 hour, allow to warm to room temperature end swir an additional 2 hours. Pour the mixture into water (100mL)and extract into ethyl ether (100mL then SOmL).- C~mbine the organic phases and dry (Na2S04). Evaporate the solvent in vccua and purify by silica gel chromatography (40~ ethyl acetate/hexane) to give the title compaund (v40mg).
Preparation of Boc-phenylene-C(0)--Phe CF
Dissolve Boc-phenylene-C(0)-Phe[OH][CFA, (0.6g) in methylene chloride (25mL) and add Dess-Martin reagent (1.8g). Stir for 48~~.hour and pour into a mixture of sodium hydrogen carbonate (O.Sg) and sodium bisulfate (1.41g) in water (25mL) and ethyl ether (100mL). Separate the organic phase and extract the aqueous phase with ethyl ether (50mL).
Combine the organic phases and dry (MgSO~). Evaporate the 2~.~~~~~
WO 92/20357 ' . PCT/LJS92/032~~

solvent invacuo and purify by silica gel chromatography (25~
ethyl acetate/hexane) to give the title compound (430mg).
Preparation of H20C-phenylene-C(0)-PhefCF~]
Dissolve Boc-phenylene-C(0)-PhefCF3] (216mg, 0.51mmo1) in ethyl acetate (50mL) and cool to 0°C. Treat with hydrogen chlaride gas for 5 minutes and stir for 3 hours. Evaporate the solvent invacuo to give the title compound 197mg).
Preparation of fCF3~Phe-G~0)°>~henylene-C(O)-Val-Fro-ValfCF2CF~]°-SEQ ID N0: 16 Suspend ~3aOC-phenylene-C(O)-PhefCF~] (175mg) in methylene chloride (xmL) and ad.d N-methylmorpholine (100uL). Cool to -22°C and add isobutylchloroformate (65uL). Stir at -22°C
for 25 minutes and add a solution of Va1-Pro-ValfCFZCF3~
(240mg) in methylene chloride (2mL) and N-methylmorpholine (60uL). Stir at -22°C for 30 minutes, allow to warm to room temperature and site for an additional 1e5 haurs:
Evaporate the solvent in vacuo to approximately lmL volume and purify by ailica gel chromatography (50~ ethyl acetate/hexane) to give the title compound (llOmg).
Example 3 CF Phi-Pro-Val-C 0 -Val-Pro--Val CF CF --SE ID NO s 17 Partition Val-Pro-ValfCF2CF3]~hyd'rochloride (2OOzng) between ethyl acetate (lOmL) and saturated sodium hydrogen carbonate (20mL}. Separate the organic phase and extract the aqueous phase with ethyl acetate (3X20mL). Cambine the organic phases, dry (MgS04) and evaporate the solvent in vacu~ to-....~lve Val-Pro-Val [ CFZCF~ ]
Partition Val-Pro-PhefCF~].hydrochloride (200mg) between ethyl aoetate (lOmL) and saturated sodium hydrogen carbonate (20mL). Separate the organic phase and extract PCT/~CJS92/~328 WO 9212tD357 the aqueous phase with ethyl acetate (3X20mL). Combine the organic phases, dry (MgSO~) and evaporate the solvent in vacuo to give Val-Pro-Phe[CF3]:
Dissolve phosgene (545mg,, 5:51mmo1) in benzene (3.5mL) and slowly add a solution of Val-Pro-Val[CF2CF3] (568mg, 1.37mmo1) in benzene (1mL). Stir at 60-75°C f~r several hours then reflux vigorously for 2 hours. Remove approximately 1/2 the benzene by distillation and cool the remaining solution in an ice bath. Add a solution of V~1-Pro-Phe[CFg] (500mg~ 1:37mmo1) in ethyl ether (2mL) and st~.r at room temperature for several hours. Evaporate the solvent in vcxcuo and purify by chromatograph~r to give the title compound.
Exam :Le 4 CFzlPhe-Pro-Val-CH2°CH2-Val-Pro-Val[CFzCF~~I--S~.ID NO: I8 Dissolve Val-Pro°Val[CFZCF3] (~68mg, 1.37manol) and Val-Pro-Phe[CFA] (500mg, 1.37mmo1) in methanol (2pmL) and add gl,yoxal (200mg of a 40~ solution in water, 1,37mmo1), sodium cyanoborohydxide (~6mg, 1.37mmo1) and 1 drop of 1~
bromocresol green in ethanol: Maintain the pH of the reaction with 1N hydrochlox'ic acid in methanol until the indicator no Iongex changes. Evaporate the solvent inUUCUo and partiti~n the residue between l~T sodium hydroxide (5mL) and ethyl acetate (lOmL). Separate the organic pha~~, dry (MgSO~ ) and evaporate the salvent in vrzcuo. Purify by chromatography to give the title compound:

wo ~zizo3s~ Pcreus~zro~a~b . _40~.
- SEI~UENCE LISTING
(1) GENERAL INFORMATION:
(i) APPLTCANT:
Angela~tro, Michael R

Bops Ptlilig~pe Doherty, Niall S

~lanllSZo ~4l~hafW T

Mehdi, Shujaath Poet, Norton P

(ii) TITLE OF INVENTION: Inhibitors of CatheFsin G aad Elastase for Presenting Conn~ctiva Tissua Degradation (iii) NUPiBER OF SEQUENCES:'18 ( iv CORRESPC~NDENC~ ADDEtESS
) (A) ADDRESSEE: Marion ~terrell Dow Inc.

(B) STREET: 2110 East Galbraith Rd.

(C) CITX: Cincinnati P. 0. Box 156300 (D) STATE: ~hi~

(E) COUNTRY; USA

(F) ZIP: q5215-6300 (v) COMPUTER READABLE'FORM:

(A? MEDIUM TYPE:.FIopPY desk (B) COMFUTER: IBM P~ comgatible ( c ) o~E~ATIta~ SYSTEM : pC-~tss/ras-DOS

(D) SOFTWARE: Patentln Release #1.0, Version X1;25 (vi) CURRENT APPLICATION DATA:

(A) APPL~C~TION IdUIM~ERt US 07/704,499 (H) FILING DATE: 23-MAY-1991 (~, CLASSIFZCATI~N:

2 5 ( v ATTORt5E3t/AGEPiT IP1FOR~iA'~ aoN r' i i i ) (A) NAME: Nesbi~t; Stephen L

(B) REtaIS'T'RATION NUMBER: 28,981 (C) REFERENCE/DOC1KET NUMBER: MO1593 ( ix ) TELEC03~lMUNICATION INFORt~IATION:

( A) TE~.EFI~OPfiE: ( 513 )' 948-7965 (B) TELEFA7Is (513) 9A8-7961.

.... - (C) ~'ELEX: 214320 ~~.~~~y;~
'CVO 92/20357 P(.'T/1JS92/0328~
-4~--(2) INFORMATION FOR SEQ Id7 NO:1:
( i ) SEQUENCE C~iAR&~CTERTSTICS:
(A) LENGTH: 8 amino acids (8) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1:

~ Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa O

(2) INFORMATION
FOR SEQ
ID NO:2:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 4 amino acids 15 (H) TY1PE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE ~'YPE: peptide (xi) SEQUENCE pESCRIPTIONSEQ ID NO:2:

xa~ xaa xaa xaa 2 ) II~IFORMATION
FOR SEQ
ID N0:
3 :

(i) SEQUENCE CHARACTERISTICS:

~5 (A) LE~TGT~3: 4 amino acids (B) TYPE: amino acid (~) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID No:3:

Ala ~waa ~r~ vai WO 92/20357 P'!v''I'/US92/032~~
-~2-( 2) INFORMATION FOR SEQ .~ID Ni0:4:
(i) sEQuENCE cH~Rn~TERISTI~s:
(A) LENGTH: 4 amino acids (R) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID N0:4:

Xaa ltaa Xaa 7(aa (2) INFORMATION
FOR
SEQ
ID NO:
r:

(i) SEQUENCE CFiARACTERISTTCS:

(A) LENGTH: 4 amino acids (B) TYF~Eamino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide ( x i SEQUENCE D1ESCRIPTIOP1 s ~EQ ID N0: 5 ) 2d Ala Ala Pro &~he (2) INFORMATION
FOR
SEQ
ID NO:6:

(i) SEQUENCE CHARACTERISTICS:

Z (A) LEN~~'H: 6 amino acids -(B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE:,peptide (ix) FEATURE:

30 (A) NPyME/REY: Modified-site (,~ ) LOCATION : 1 O .....(D) THER INFORMATION: /note= "Xaa is a valine analog having a CF2CF3 gr~up replacing the hydroxy of the terminal c~rboxy."

(ix) FEATURE:

(A) NAME/KEY: Modified-site .

~~ ~~~;:~~z WO 92/20357 PC.'T/US92/032~~

(B) LOCATION: ~
(D) OTHER INFORMATION: /note= "Xaa is a valine ~na~ag having a -L1-Ph-L2- group replacing one of the hydrogens on the amino terminus wherein Ll and L2"
(ix) FEATURE:

(A) NAME/KEY: Modified-site . (8) LOCATION: 3 (D) OTHER INFORMATION: /note= "(eont) are each a carbonyl wherein L1 is bhund to the peptide fr~agsient containing the modified sites at locations l-2 "

( ix) FEATI3RE:

(A) NA~IE/KEY: Modified-site 4 ~ ) Lo~ATIOr~ : ~

(D) OTHER IN~'ORMATTON: /notes "(cont)and L2 is bound to the peptide fragment containing the m~dified sites at locations 4-6'and Fh as a p--phenylene group."

( i x FEATURE
) (A) NAME/KEY: Modified-site (B) LOCATIQN: f (D) OTHER INFORMATION: /not~.e= "Xaa is a phenplalmnine analog having a -CF3 group replacing the hydroxy of the terminal cartaoxy"

(y~l~ ~EQIIENCE DESCRIP'TIO'N: SEQ I~ NO:6:' Xaa Pro Xaa Val Pro xaa (2) INFORMATION
FOR
SEQ
ID NO:7:

(i) SEQUENCE CHARACTERISTICS:

(p~) LENGTH: 4 amino acids (B) TYFE:' amino acid (D) TOPOLOGY; linear ~ ii MOLEC1JLE ~'YPE: Peptide ) (xi) SEQUENCE DESCRIPTION: SEQ ID N0:7:

...

X~~
xaa Xaa xaa W~ 92/20357 FC'T/U~92/U32~~

(2) INFORMATION ~'OR SEQ ID NO:B:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: ~ amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ LD N0:8:
Xaa Xaa Xaa xaa Z
(2) INFORMATION FOR SEQ ID N0:9:
(.i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 4 amino acids (B) TYPE: amino,acid 1 5 (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPT30N: ~EQ TD ~d0.9:
Xaa Xaa Xaa Xaa (2) INFORMATION FOR SEQ ID P~O:10:
(i) SEQUENCE CHARAC'~ERISTICS:
(A) LENGTH: 4 amino acids 2S (B) TYPE: amino acid (D) TOPdLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:10:
3 ~D ._. .
xa~.Xaa Xaa Xaa WO 92/20357 Pt; I'/tJ592/03288 (2) INFORMATION FOR SEQ ID NO:11:
(i) SEQUENCE CRARACTERISTICS:
(A) LENGTH: 4 amino acids ($) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:11:

xaa xaa xaa Xaa ~. 0 1 (2) INFORMATION
F'QR
SEQ
ID N0:12m (i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 4 amino acids (B) TYPE: amino acid (D) T~POLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ITp N0:12:

2 0 xaa Xaa xaa xsa (2) INFORMATION
FOR
SEQ
ID NO:13:

( SEQUENCE ,CHARACTERISTICS
) i (~) L~~1GT~: 4 amino acids (B) TYPE:: am~.no acid 25 (D) TOPOLOGY: lineal (ii) MOLECULE TYPE: PePti~a (xi) SEQUENCE DESCRIPTION: SEQ I17 NO:1~:

xaa xaa xaa xaa WO 12/20357 PC'~'liJS92/032~8 (2) INFORMATION FOR SEQ ID N0:14:
(i) SEQUENCE CHARACTERISTICS;
(Aj LENGTH: 4 amin~ acids (H) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID N0:14:

Xaa Xaa Xaa Xaa io 1 (2) INFORMATION
FoR
SEQ
ID N0:15:

(5.) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 4 amino acids (B) TYPE: amino acid (D) TOPOLOGY; linear 1 ~

(ii) MOLECULE TYPE: peptide (x~tj SEQUENCE DESCRIPTION: SEQ ID N01~:

Xa~ Xas: Xaa Xa3' 2 ) INFOFtMP.TION
FOR
SEQ
ID IdO:16 (i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 4 amino acids (H) TYPE: amines acid (~j TOP~LOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE;

(A) NF~1~IE/KEY: Modified-site 3 0 (~) LOCATION: ~

..... (D) OTHER IHFOFtMATION: /notes "Xaa is a va~.ine ahalog having a -CF2C~'3 group rap7.acing the hyd~oxy of the ~,erminal c~rbox~.' i x ) F~ATITRE

(A) NAI~tE/KEY: Modified-site (H) LOC1~TION: ' 3 3~

WO 92/2037 PCT/US92/032~~
°47-(D) OTHER INF~RMATION: /note= "xaa is a valine analog having a -Ll-Ph-L2-group replacing one of the hydrogens on the amino terminus wherein L1 and L~
(ix) FEATURE:

(A) NAME/KEY: Modified-site (8) LOCATION: 3 (D) OTHER INFORMATION: mote= "(contj are each a carbonyl wherein Ll is bound to the peptide fragment containing the modified sites ~t locations 1-2 (ix) FEATURE:

(A) NAME/KEY: Modified-site H ) LOCA'f I ON : 3 (D) OTHER INFORMATION: /n~te= ~(cont)and L2 is bound to the peptide fragment containing the modified site at location ~4 arad ph is ~ p-phenylene gr~up."

(ix) FEATURE:

(A) NAME/KEY: Modified-site (U) LOCATION: 4 (D) OTHER INFORMATION: ,/note= "xaa is a phenylalanine 1~ analog having a -CF3 gr~up replacing tkae hydroxy of the terminal carboxy."

(xi) gE~UENC~ DESC~IPTI~x: sEQ ID NO:ls:

Xaa Pro Xaa Xaa ( 2 ) INFoRI~lATION
FCJR
sEQ
ID No:
l7:

( l ) sEQUE~rc~ cHA~cTERrsxacs:

(A) LENGTH: 6'amino acids (H) TYPE: amino acid (D) TOP~L~GY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:

(A) NAME/%EY: Modified-Site (B) LOCATION: l ... (DD oTxER INFORM~.Tao~t: Jnobe-- "xaa is a valine analog hawing ~ -OF~~F3 group replacing the hydroxy of the terminal carboxy."

( FEA~tIRE
x ) l (A) NAM~/KEY: Modified-site-(B) L~DC~TI~PI: 3 (D) OTHER INFORMATION: /note= "Xaa is a valine ainalog i ~i~ i,i wo ~ziza3s7 pc-rrus~zm3zs~

having a -C(O)- group replacing one of the hydrogens on the amino terminus."
(ix) FEATURE:
(A) NAME/ICEY: Modified-site (B) LOCATION: 6 i~%~) OTHER IidFORMATION: /note= "Xaa is a phenylalanin~
analog having the -CF3 group replacing the hydsoxy of the terminal carboxy."
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: I7:
Xaa Pro Xaa Val Pro Xaa (2) INFORMATIO?t FOR SEQ ID IdOelB:
(i) SEQUErICE CHARACTERISTICS:
(A) LE~'GTH: 6 amino acids (H) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:
(~,) NAME/icEY:' Modified-site (~j LcxATroN: a ~0 (D) OTHER INfFORMATIOPI: /notes "Xaa is a valine analog having a -CF2~F3 group,replaciag the hydroxy of the terminal ~arboxy."
(ix) FEATURE:
(A) ~IAME/K~Y: Madified-site H ) LOCB~TIOPJ : 3 (D) OTHER INFORMATIgH: /note= °'Xaa is a valine analog having a ~(e~i2jn+2~ group saherefn n is 0 replabing one of the hydrogens on the amino terminus,"
(ix) FEATURE:
(A) NAME/ICEY: Modified-site (B) LOCATION: 6 (D) OTFTEFt IIdF~RMATION: /note= "Xaa is a phenylalanine ..... ~ analog having a -CF'3 group replacing the taydroxy of the terminal carboxy."
(xi) SEQUENCE DEBCRIPTIOPt: SEQ ID NO:l~:
Xaa Pro Xaa Val Pro Xaa

Claims (15)

WHAT IS CLAIMED IS:

1. ~A compound of the formula wherein P1 is Ala, bAla, Leu, Ile, Val, Nva, bVal, Met, Nle, Gly, or Sar;
P1' is Ala, bAla, Leu, Ile, Vai, Nva, bVal, Met, Nie, Phe, Tyr, Tyr(Me), Ala(3pyr), Ala(4pyr), Trp, or Nal(1);
P2 is Pro, Ind, Ala, bAla, Leu, Ile, Val, Nva, bVal, Met, Nle, Phe, Tyr, Tyr(Me), Ala(3pyr), Ala(4pyr), Trp, or Nal(1);
P2' is Pro, Ind, Ala, bAla, Leu, Ile, Val, Nva, bVal, Met, Nle, Gly, Sar or is absent;
P3 is Lys, Arg, Pro, Ind, Ala, bAla, Leu, Ile, Val, Nva, bVal, Met, or Nle;
P3' is Ala, bAla, Leu, Ile, Val, Nva, bVal, Met, Nle, Gly, Sar or is absent;
P4 is Ala, bAla, Leu, Ile, Val, Nva, bVal, Met, Nle, or is absent;
P4' is Ala, bAla, Leu, Ile, Val, Nva, bVal, Met, Nle, Gly, Sar, or is absent;
L is a -C(O)-phenylene-C(O)group;

EIM and CGIM are each independently selected from the group consisting of -C(O)C(O)R, -CF2CF3, -CF3, -CF2H, -CO2R3, -CONHR3, -CF2CHR3C(O)NHR, -H, alkyl, aryl, aralkyl, -C(O)R, wherein R3 is H, alkyl, phenyl, benzyl, R is OH or alkoxy or a pharmaceutically acceptable salt thereof.

2. A compound of claim 1 wherein CGIM and EIM are each independently either a -CF3 or -CF2CF3 group.

3. A compound of claim 1 wherein P1 is norvaline or valine;
P1' is phenylalanine;
P2 is proline;
P2' is proline or is absent;
P3 is isoleucine, valine, or alanine;
P3' is alanine, valine or is absent;
P4 is alanine or is absent; and P4' is alanine or is absent.

4. A compound of claim 3 wherein CGIM and E1M are each independently selected from a -CF3 or -CF2CF3 group.

5. A compound of claim 1 wherein -P4-P3-P2-P1 (SEQ ID NO: 2) is a -Ala-Ala-Pro-Val-(SEQ ID NO: 3); -Lys(2CBz)-Pro-Val-; or -Val-Pro-Val- group.

6. A compound of claim 2 wherein -P4-P3-P2-P1 (SEQ ID NO: 2) is a -Ala-Ala-Pro-Val-(SEQ ID NO: 3); -Lys(2CBz)-Pro-Val-; or-Val-Pro-Val- group.

7. A compound of claim 1 wherein -P4'-P3'-P2'-P1' (SEQ ID NO: 4) is a -Ala-Ala-Pro-Phe (SEQ ID NO: 5); -Val-Pro- Phe-; or -Phe- group.

8. A compound of claim 2 wherein -P4'-P3'-P2'-P1' (SEQ ID NO: 4) is a -Ala-Ala-Pro-Phe (SEQ ID NO: 5); -Val-Pro-Phe-; or -Phe- group.

9. A compound of claim 1 which is

10. Use of a compound according to any one of claims 1-9 for preparing a medicament for treating gout or rheumatiod arthritis in a patient in need thereof.

11. Use of a compound according to any one of claims 1-9 for preparing a medicament for treating neutrophil associated inflammatory disease in a patient in need thereof.

12. Use of a compound according to any one of claims 1-9 for preparing a medicament for treating conditions associated with connective tissue degradation in a patient in need thereof.

13. Use of a compound according to any one of claims 1-9 for treatment of gout or rheumatoid arthritis in a patient in need thereof.

14. Use of a compound according to any one of claims 1-9 for treatment of neutrophil associated inflammatory disease in a patient in need thereof.

15. Use of a compound according to any one of claims 1-9 for treatment of conditions associated with connective tissue degradation in a patient in need thereof.