KININOGENASE INHIBITORS

Abstract: Kininogenase inhibitors, optimally not exceeding the size of a hexapeptide, represented by (II), wherein A and B = amino acyl (including amino acyl analogue) the same or different forming a dipeptide group the amino acid of A carrying a terminal group and being any amino or imino-acid residue (but preferably of D-configuration) and of B being a lipophilic amino-acid residue of D- or L-configuration but not proline or a proline analogue, or a conformational analogue of said dipeptide group wherein the peptide link is replaced by -CH2-NH- ('reduced'), -CH(OH)-CH2- ('hydroxy'), -CO-CH2 ('keto'), -CH2-CH2- ('hydrocarbon') or other conformational mimic of the peptide bond and in (III) the side chain R<1> is that of a basic amino acid or amino acid analogue (preferably of L-configuration), and R is H or lower alkyl(C1-C4) or C< alpha > or the peptide link comprising -N(R)- is replaced leading to a conformational mimic as above; Y = a binding enhancing or carbonyl activating group preferably selected from H (when A or B must be cyclohexylalanine, preferably D if at A or L if at B) or alkyl (C1-C20) or fluoroalkyl (C2-C12); substituted oxymethylene; thiomethylene; sulphoxymethylene; sulphonylmethylene; aminomethylene; hydrazino-methylene; -CH2-Het (where Het = a substituted or unsubstituted heterocycle); substituted amino (but when the resulting compound is a secondary alkylamide B must not be phenylalanine); an amino-acid group or its ester or amide; a carboxylic secondary amide or primary amide, when B must be cyclohexylalanine or adamantylalanine or other bulky lipophilic, non-aromatic amino-acid (not Ala Leu Ile Val Nva Met Nle Phe Tyr Trp Nal (1)); tertiary-carboxamide; carboxy-alkyl group or its ester or amide or amino acyl derivative.

Description: KININOGENASE INHIBITORS FIELD OF INVENTION The invention relates to enzyme inhibition and to treatment of disease. BACKGROUND - KININS Kinins are natural vasoactive peptides liberated in the body from high molecular weight precursors (kininogens) by the action of selective proteases known as kininogenases. There is evidence for the involvement of kinins in the following pathological states: (a) Conditions associated with vasodilatation and hypotension, e.g. septic, anaphylactic and hypovolaemic shock; carcinoid syndrome and dumping syndrome (b) Conditions involving inflammation, e.g. acute arthritis, pancreatitis, local thermal injury, crush injury and brain oedema (C) Conditions involving bronchoconstriction, especially for example the initial, acute allergic reaction in asthma (d) Allergic inflammation, particularly allergic rhinitis and conjunctivitis, together generally known as hay fever, and the bronchial inflammation and consequent occlusion found in the non-acute but serious and even fatal inflammatory phase of asthma. The kinins ( bradykinin, kallidin and Met-Lys-brε kinin) are potent medic rs of inflammation. Their r .n actions are as follows: (a) They increase capillary permeability which leads to exudate formation and oedema (b) They are potent vasodilators in arterioles and therefore reduce blood pressure and increase blood flow (c) They induce pain (d) They contract bronchial smooth muscle (e) They activate phospholipase A 2 and thus stimulate the biosynthesis of prostaglandins (PCs) which mediate some of their actions. In regard to prostaglandins, it may be noted that certain actions of kinins, particularly pain and vascular permeability above, are potentiated by PCs, although PCs themselves do not cause pain nor do they induce vascular permeability at the concentrations found in inflamed tissue. PCs therefore act as either mediators or potentiators of kinins. In spite of the above knowledge of kinins and their actions, relatively little attention has been paid to reduction of their action. In asthma treatment for example clinical attention is primarily directed to the acute bronchoconstrictive reaction, for which there are effective drugs. Deaths continue to occur from the gradually developing bronchial occlusion, and at present not only are there no clinically effective inhibitors of kinin release available but the concept of kinin release inhibition, at least in treating allergic inflammation, appears to be new. The only substance that is in fact a kinin release inhibitor and has attained clinical significance is aprotinin ('Trasylol' , Bayer, trade mark) , a proteinase inhibitor isolated from bovine tissues (lungs, lymph nodes and pancreas). It is a strongly basic protein (pi = 10.5) of MW = 6,500 comprising a single peptide chain of 58 residues. However, aprotinin is primarily a trypsin inhibitor (Ki = 10~ 13 M) and is some 10 6 -times less active against kinin release. It has been found marginally beneficial in acute pancreatitis, a serious condition, where it inhibits the activation by trypsin of zymogens of pancreatic serine proteinases, and in traumatic - haemorrhagic shock. Aprotinin has to be administered parenterally, and it frequently produces a painful reaction at the injection site. BACKGROUND - KININOGENASES The kininogenases are serine proteinases, that is to say proteinases in which the hydroxy group of a serine residue is the nucleophile involved in forming the substrate transition state. They liberate the kinins (bradykinin, kallidin) from the kininogens by limited proteolysis. There are several kinds of kininogenase:- (a) Tissue kallikrein (TK, also called glandular kallikrein GT or urinary kallikrein UK) which is found in the pancreas, salivary glands, intestines, kidney and urine. It has MW = 30,000 and acts preferentially on low molecular weight kininogen (LMWK) to release the kinin kallidin (KD). Tissue kallikrein has no potent and fast acting endogenous inhibitor present in plasma. (b) Plasma kallikrein (PK) occurs in plasma as an inactive zymogen which is activated by Factor Xlla, and is part of the intrinsic coagulation cascade. It has MW = 100,000 and its preferred substrate is high molecular weight kininogen (HMWK) from which it releases bradykinin (BK). Plasma kallikrein is rapidly and effectively inhibited in plasma, by endogenous inhibitors known as cl-inactivator and α 2 -macroglobulin. (c) Mast cell tryptase which, while not as active as the kallikreins in kinin release, we have found to occur in large amounts in the mast cells of the lung tissue of asthmatics. BACKGROUND - KININOGENS The kininogens which are the natural substrates for the kininogenases (they act also as potent inhibitors, Ki approx. 10 ~ - ■ - ■ , of cysteine proteinases such as cathepsins B, H and L, calpaih -and papain) occur in two types: (a) Low molecular weight kininogen (LMWK) with molecular weight in the range 50,000 - 70,000 depending on species of origin and degree of glycosylation. (b) High molecular weight kininogen (HMWK) with molecular weight in the range 88,000 - 114,000 which, in addition to serving as an alternative precursor of kinins and a cysteine proteinase inhibitor, also plays an obligatory role with plasma kallikrein in the initiation of the intrinisic coagulation cascade. The two kininogens, whose RNA's are transcribed from the same gene, have identical primary sequences throughout the N- terminal or heavy chain (H-chain) region, the kinin region and the first twelve amino acids of the C-terminal or light chain (L-chain) . At this point their structures diverge, HMWK having a longer L-chain (MW approximately 45K) than LMWK (4.8K) . The cleavage of human HMWK by plasma kallikrein is for example shown schematically in Fig. 1, with details of the sequence at the cleavage sites in Fig. 2 and a more detailed sequence in Fig. 3 where the conventional numbering of residues adajcent to a cleavage site is shown for cleavage site I. After excision of one or other kinin sequence, the H- and L-Chains are held together by a single disulphide bridge:- HMWK -100K PK-100K BK -IK Figure 1. Cleavage of HMWK by PK: Overall scheme PK PK + + TK • Cleavage site II TK + -Ile-Ser-Leu-Met • Lysj-Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Ar Ser-Ser-Arg-Ile-Gl 380 390" Bradykinin ' Cleavage Cleavage site I site HI _ Kallidin — Figure 2. Cleavage of human kininogens by PK and TK: Details of sequence PK.TK P- P P 5 4 3 p i P' 3 P P 5 ■ Phe-Ser-Pro-Phe-Arg ■ Ser-Ser-Arg-Ile — Gly- 385 389 390 394 Figure 3. Sequences flanking cleavage site I in human HMWK As shown, plasma kallikrein and tissue kallikrein act at a single site to free the kinin C-terminal site, cleaving between residues 389 and 390, but at sites one residue apart, either side of residue 380, to free the N-terminal of bradykinin (by PK) or kallidin (by TK). The role of PK and HMWK as clotting factors in the intrinsic cascade does not involve the enzymatic release of kinins. However many of the effects of PK and all those of TK do involve such release, being mediated by the kinins released from the respective substrates HMWK and LMWK through selective proteolysis. INDICATIONS The main clinical indications for kininogenase inhibitors are inflammatory conditions, particularly allergic inflammation (e.g. asthma and hay fever). A fuller list of indications is given below: (1) Allergic inflammation (e.g. asthma, rhino-conjunctivitis [hay fever], rhinorrhoea, urticaria (2) Inflammation (e.g. arthritis, pancreatitis, gastritis, inflammatory bowel disease, thermal injury, crush injury, conjunctivitis ) (3) Smooth muscle spasm (e.g. asthma, angina) (4) Hypotension (e.g. shock due to haemorrhage, septicaemia or anaphylaxis, carcinoid syndrome, dumping syndrome) (5) Oedema (e.g. burns, brain trauma, angioneurotic oedemε whether or not as a result of treatment with inhibitors of angiotensin converting enzyme) (6) Pain and irritation (e.g. burns, wounds, cuts, rashes, stings, insect bites) STATEMENT OF INVENTION In one aspect the invention provides a method of treatment (including prophylactic treatment) of an inflammatory or other condition set out in the indications above, particularly, an allergic inflammatory condition, wherein an effective amount of a peptide or peptide-analogue kininogenase inhibitor is administered topically or systemically to a patient suffering from or at risk of the condition. It is believed that for optimum activity, ad inistrability and stability in the body the compounds should not exceed the size of a hexapeptide, that is to say should not comprise more than six amino acid or amino acid analogue residues; the presence of further residues, particularly in a pro-drug from which residues are cleaved in the body to give the compound primarily exerting the desired effect, is however not excluded. Particularly, the invention provides a method of treatment of the allergic inflammatory phase of asthma, wherein an effective amount of a kininogenase inhibitor such as a mast cell tryptase inhibitor is administered topically or systemically to a patient suffering from or at risk of the condition. The invention extends further to a method of preparation of a medicament for the topical or systemic treatment (including prophylactic treatment) of conditions as above particularly for allergic inflammatory conditions and especially for asthma as above, wherein a kininogenase inhibitor is associated with a pharmaceutically acceptable diluent or carrier to constitute said medicament. In the above, the kininogenase inhibitor is conveniently but not essentially of the novel kind now described in which in another aspect, without limitation to any particular clinical indication, the invention provides synthetic, low molecular weight compounds that selectively inhibit kininogenases and thus block the release of kinins from kininogens. The inhibitors are peptide analogues, desirably (as above) not exceeding the size of a hexapeptide in terms of amino acid or analogue residues, based on the known amino acid sequence of the kininogens at cleavage site I, which analogues have sufficient similarity to the cleavage site sequence to bind to the active site of the kininogenase but are not hydrolysable and therefore remain bound, inactivating the enzyme. The inhibitors are essentially of the structure below, in which A represents the P 3 residue, B the P 2 residue, C the P- [ _ residue and Y a carbonyl-activating or binding group the structure being:- A - B - C - Y I where A, B and C are amino acyl or amino acyl analogue groups linked by peptide bonds or conformational analogues thereof giving a peptide mimic. Other residues in addition to these essential ones may of course be present, including amino acyl or amino acyl analogue residues. In more definitive terms the compounds are represented by wherein A and B = amino acyl (including amino acyl analogue) th same or different forming a dipeptide group th amino acid of A optionally carrying a termina group (other than hydrogen) and being any amin or imino-acid residue (but preferably of D configuration) and of B being a lipophili amino-acid residue of D- or L-configuration bu not proline or a proline analogue, or conformational analogue of said dipeptide grou wherein the peptide link is replaced b -CH 2 -NH- ('reduced'), -CH(OH)-CH 2 - ('hydroxy'), -CO-CH 2 - ('keto'), -CH 2 -CH 2 - ('hydrocarbon') or other conformational mimic of the peptide link and in:- R ' 0 the side chain R 1 is that of a basic amino aci or amino acid analogue (preferably of L configuration) and R is H or lower alkyl ( C^ C 4 ) or C α or the peptide link comprisin -N(R)- is replaced leading to a conformationa mimic as above. For example C α may be replace by nitrogen. = a binding enhancing or carbonyl activatin group for example selected from H (but only i A or B is cyclohexylalanine, preferably D if a A or L if at B) or alkyl C-^ - C 20 ) or fluoroalkyl (C 2 - C 12 ); substituted oxymethylene; thiomethylene; sulphoxy- methylene; sulphonylmethylene; aminomethylene hydrazino- ethylene; -CH 2 -Het (where Het = a substituted or unsubstituted heterocycle) ; substituted amino (but when the resulting compound is a secondary alkylamide B must not be phenyl-alanine) ; an amino-acid group or it ester or amide; a carboxylic secondary amide or primary amide, when B must be a bulky lipophilic, non-aromatic amino-acid e.g. cyclohexylalanine, adamantylalanine (not Ala Leu lie Val Nva Met Nle Phe Tyr Trp Nal (1)) ; tertiary-carboxamide; carboxy-alkyl group or its ester or amide. In the above context substituents are suitably common functional groups that increase binding affinity to the enzyme and/or improve pharmacological properties. Further in considering conformational analogues or mimics a dipeptide mimic is a structure containing non-natural amino acid (amino acid analogue) residues or which is non-peptidic and which in I holds the side-chains of A and B or B and C or all of them in a conformation similar to that present in the parent peptide when bound to the active site of the enzyme. It may also contain features favourable for other interactions with the enzyme, e.g. hydrogen bonding. A mimic may be chosen from the published work on such analogues. For example, the following are mimics of the dipeptide DPro-Phe (Ph may be replaced by -CH 2 Ph):- (i) ' R educed' mimic (ii) 'Hydroxy' or (iii) 'Keto* or hydroxyethylene mimic ketomethylene mimic (iv) (v) (vi) ( ' Hydrocarbon ' mimic) Figure 4, DPro-Phe mimics The preferred compounds represented by the above general formula are now considered. Preferred residues for A are imino-acids, (e.g. D-proline or an analogue of proline e.g. pipecolinic acid, azetidine carboxylic acid etc.); lipophilic amino acids (e.g. DPhe, DCha, DChg) ; strongly basic amino acids (e.g. D-Arg or a guanidinophenylalanine) and for B they are L-Phe, L-Cha, L-αNal, L-Tal, L-(4F)Phe L-(NMe)Phe or other substituted phenylalanines. A and B may also be the N-alkyl (C 1 -c 4 ) or Cα-alkyl ( -^-C j e.g. methyl, benzyl) analogues of these amino acids. Suitable terminal groups for A include lower alkyl (preferred) or acyl (not excluding amino acyl), alkyl sulphonyl (straight chain or branched or cyclic), amino-alkyl, carboxy alkyl, hydroxy alkyl or any other common protecting group encountered in peptide chemistry. Groups suitable as group Y are specific to the present invention in that they are part of the structure giving the required binding to the active site and are not merely non- interfering end groups. They form a binding group which increases affinity to the enzyme and/or a group which activates the adjacent carbonyl by rendering it more electrophilic. Specific groups are included in the following formula: where in a peptide link to residue B the cx-nitrogen may be free or substituted for example by methyl or other C-^-C^ alkyl and thus R 1 and R are as before but particularly R 1 = 3-guanidinopropyl or other guanidinoalkyl group or an amidino- alkyl or a inoalkyl group, also para- or eta substituted guanidino or amidino-benzyl or protected forms of the above (the basic nitrogens may also be alkylated e.g. with Me, Et) , and where Y * = groups as given below, first in more general terras and then in terms of more detailed preferences, subject in both cases to the provisos expressed in defining the compounds of the invention earlier. The detailed preferences are given in groups under roman numerals, which are also indicated, in brackets, with the first listing which is:- (I) Y = H (representing aldehydes) or alkyl including fluoroalkyl (representing ketones) (II-II -IV) Y = -CH 2 Q where Q = -OR 2 or -SR 2 or -SOR 2 or -S0 2 R 2 or / -NHR * or - N or (note that -DJ represents a ring in which D is an atom of that ring) wherein R 2 , R 4 and R 5 are as below R- / ( V-VI ) Y = -CH CHR° CON or -CH-, CHR 6 CO-D) wherein R 4 , R D and R° are as below (VII-VIII) Y = amino acyl or a group forming a substituted amide or hydrazide (IX-X-XI) Y = a group forming an α-keto amide e.g. - COR 9 or -CO-D or R" -CON ^ R and in which further:- R **** = alkyl or substituted alkyl including aarryyll oorr aarr^yl alkyl and -CH 2 R 3 where R 3 fluoroalkyl R 4 and R 5 the same or different but not both hydrogen = H or ^±-C 2 Q alkyl (which may be further substituted) , acyl or alkyl sulphonyl -ΕJ is a heterocyclic ring (D = nitrogen or carbon in Group IV and N in Groups VI and X) optionally unsaturated and optionally with further hetero atoms and substituents R 6 = hydrogen, alkyl, hydroxyalkyl, aminoalkyl, alkylaminocarbonyl R 9 * = -NH as such or alkylated, or amino acyl The listing of more detailed preferences, again within the provisos expressed earlier, is:- Group I Y = H; alkyl including branched alkyl (C- j ^C^); aryl alkyl; or cycloalkyl (C-^-C^); perfluoroalkyl or partially fluorinated alkyl (C 2 -C 12 ); [ e - < 3- γ = Me ' -CH(CH 2 CH 2 CH 2 CH 3 ) 2 ; -CH(CH 2 CH 2 CH 2 CH 3 )CH 2 -cyclohexyl; —CH CF C CF-5; ] . Group II Y = -CH 2 QR 2 where Q = O, S, SO, S0 2 , NH and where R 2 = Alkyl, branched alkyl or acyl (C 1 -C 1 ); or cycloalkyl (C^C^); or aryl or aryl alkyl; or -CH 2 -R 3 where R 3 = perfluoroalkyl or partially fluorinated alkyl, branched or not (C- ] _-C 12 ). Group III R 4 R 4 , R 5 the same or different = alkyl, branched alkyl, cycloalkyl, acyl, alkylsulphonyl, carboxyalkyl (the carboxyl group may be further derivatized to form an ester or amide with an amino-acid or dipeptide) , carbamoyl, suiphamoyl, N-dialkylamino-, arylalkyl, haloalkyl including fluoroalkyl, cyanoalkyl, alkoxyalkyl. hydroxyalkyl, mercaptoalkyl, aminoalkyl and derivatives thereof e.g. esters, amides and thioesters; or one of R 4 or R 5 = hydrogen Group IV Y = -CH - 9 where D = nitrogen or carbon and -Dj is a saturated or unsaturated heterocyclic ring or a bicyclic ring system, each is 5 - 8 membered, where there may be other hetero- atoms (N, S, 0) and carbons or nitrogens may optionally be substituted by alkyl, branched alkyl, cycloalkyl, carboxyalkyl, carboxy (attached to carbon), amino, alkoxy, alkoxymethyl or (carbon) as carbonyl or other groups beneficial for interaction with the enzyme. Group V Y = -CH 7 CH(R 6 )C0NR 4 R 5 R 4 , R 5 as defined in Group III R 6 _ = hydrogen lower alkyl, branched alkyl, cycloalkyl, hydroxyalkyl, amino-alkyl, alkylaminocarbonyl. Group VI as defined in Group V and - D) as defined in Group IV ( but with D = N) Group VII Y = an amino-acid residue or any amide (secondary or tertiary) or ester of that residue, L or D configuration. Preferred residues are of lipophilic amino-acids e.g. norleucine, cyclohexylalanine, homocyclohexylalanine , cyclohexylglycine , ter: butylglycine . Group VIII R = H (when however B is not phenylalanine unless R 8 is carboxylalkyl or derivatized carboxylalkyl) ; or alkyl, branched alkyl (C χ - C 12 ) , cycloalkyl (C 1 -C 20 ) carboxyalkyl or bis ( carboxyl ) alkyl , which may be derivatized at the carboxyl group to form an amide e.g. with an amino-acid (preferred is arginine) or a substituted amine; N'-dialkylamino; N'-alkylamino-; R 8 = R the same or different but excluding H. Group IX Y = -CO-R 9 but only if B is a bulky non-aromatic lipophilic amino acid or its N.? alkyl (C-. - C 4 ) derivative (e.g. cyclohexylalanine but excluding Ala, Leu, lie, Val, Nva, Met, Nle, Phe, Tyr, Trp, Nal(l) and their N-methyl derivatives) where R 9 = NH_, N'-alkylamino (where the alkyl groups include branched and/or cycloalkyl); an amino-acid residue. Group X -D) as defined in Group IV (but D = N) Group XI Y = -CO-NR 4 R 5 R ,R 5 as defined in Group III, but not H The following examples illustrate the invention. The are given in the form of:- Nine tables of compounds with reference number, structure, molecular "-on as determined by FAB (fas atom bombardment) spectrometrv and class of compoun (the same as the 'groups' referred to earlie herein) Eight detailed examples of synthesis Twelve synthesis schemes, as referred to in th detailed examples Table of abbreviations Description of in vitro tests of inhibition o kininogenases and in vivo tests of efficacy agains asthma All structures of intermediates were verified by NMR. TABLE 1 Me-DPhe Cha Arg-H Thiomethylene Analogues and Sulphomethylene Analogues [M+H] + Class H-DPro Phe Arg CH^CH^Me H-DPro Phe Arg CH 2 S n Bu H-DPro Phe DLArg CH 2 S0 2 π Bu TABLE 3 Ethers [M+H] + Class H-Pro Phe Arg CH 2 OCH 2 CF 3 Boc-DPro Phe DLArg CH 2 OCH 2 CF 3 H-DPro Phe DLArg CH 2 OCH 2 CF 3 H-DPro Cha DLArg CH 2 OCH 2 CF 3 H-DPro Phe Arg CH 2 OCH(Me)CF 2 CF 2 CF 3 H-DPro Phe Arg CH 2 OGH 2 CF 3 H-DCha Phe Arg CH 2 OCH 2 CF 3 H-DArg Phe Arg CH 2 OCH 2 CF 3 H-DChg Phe Arg CH 2 QCH 2 CF 3 H-DPro Phe Arg CH 2 O(CH 2 ) 5 CH 3 H-DPro Phe Arg CH 2 OPh TABLE 4 Aminomethylene and related analogues [M+H] + Class H-DPro Phe Arg CH 2 N[(CH 2 ) 5 Me] 2 600 m H-DPro Phe DArg CH 2 (RS)Tha 556 IV H-DPro Phe Arg CH 2 (RS)Tha 556 IV H-DPro Phe Arg CH 2 l-Pip-3-(RS)CO 2 Et 572 IV H-DPro Phe DLArg CH 2 l-Pip-3-(RS)CH 2 O(CH 2 ) 3 Me 586 IV H-DPro Phe DLArg CH 2 l-Pip-3-(RS)CH 2 NEt 2 585 IV H-DPro Phe DLArg CH 2 l-Pip-3-CONEt 2 599 IV H-DPro Phe DLArg CH 2 l-Pip-3-(RS)CONrPBu 599 IV H-DPro Phe DLArg CH 2 l-Pip-3-(RS)CON( n Bu) 2 655 IV H-DPro Phe DLArg CH 2 l-Pip-4-(l'-Pip) 583 IV H-DPro Phe Arg CH 2 (NMe)AhaNH n Bu 615 IV H-DPro Phe Arg CH 2 -Abn 540 IV H-DPro Phe Arg CH j -Hyp CPB^NHEt 629 IV H-DPro Phe Arg CH 2 -Sar Pro NHEt 628 in H-DPro Phe Arg O HyptCPBu^Bu 644 IV H-DPro Phe DLArg CH 2 Pic NEt 2 641 IV H-DPro Phe DLArg CH 2 t DHyp(O n Bu) B O n Bu 644 IV H-DPro Phe DLArg CH 2 Pro B CONEt 2 599.9 IV H-DPro Phe DLArg CH 2 l-Pip-3(RS)CO 2 H 544 IV H-DPro Phe Arg CH 2 l-Pip-3(RS)CH 2 CONEt 2 613 IV H-DPro Phe Arg CH 2 N(CH 2 Ch)(CH 2 ) 5 Me 612 in H-DPro Phe DLArg CH 2 N(Oc) 2 657 m H-DPro Phe Arg CH 2 N(Et)Ch 542 m H-DPro Phe Arg CH 2 N(Me)(GH 2 ) 4 NH 2 517 m H-DPro Phe Arg CH 2 N(Me) n Bu 502 in H-DPro Phe DLArg CH 2 N n Bu 2 544 m H-DPro Phe DLArg 594 m H-DPro Phe DLArg CH 2 NPBu](CH 2 ) 3 CONH 2 573 m H-DPro Phe DLArg CH 2 NI n Hex](CH 2 ) 7 NH 2 629.5 m H-DPro Phe DLArg 601 m H-DPro Phe DLArg OT 2 N[ τ Ηex](CH 2 ) 3 NH 2 573 m H-DPro Phe DLArg CH^FBuKO^Ph 620 in con . Aminomethylene Ketones and related analogues [M+H] + Class Phe DLArg CH 2 N[ π Hex](CH 2 ) 6 CONH 2 643 πi Phe DLArg CΗ 2 NFΗex](CH 2 ) 6 NH 2 615 in Phe DLArg CH 2 N[ π Hex](CH 2 ) 7 OH 630 πi Phe DLArg CH 2 NPHex](CH 2 ) 7 NHAc 671 m Phe DLArg CH 2 N[ n Hex](CH 2 ) 6 CONHEt 671 πi Phe DLArg CH 2 N[ n Hex](CH 2 ) 6 CO 2 Me 658 ΠI Phe DLArg CH 2 NPHex](CH 2 ) 6 CO 2 H 644 ΠI Phe DLArg CH 2 N[ π Bu](CH 2 ) 3 CONEt 2 629 ΠI Phe DLArg 601 ΠI TABLE 5 Keto Isostere Containing Analogues [M+H] + Class 40 H-DPro Phe DLArg^Gly Pro-NHEt 599 VI 11 H-DPro Phe Arg^Gly Pro-NHEt 599 VI 12 H-DPro Phe Arg^Gly Arg-NH 2 530 V 13 H-DPro Phe DLArg^Gly Ala-NH 2 545 V 14 H-DPro Phe DLArgKQy Aha-NH 2 587 V 15 H-DPro Phe DLArgSGly Aha-NI-FBu 643 V TABLE 6 Substrate Analogues . Substrate Analogues [M+H] + Class TABLE 7 Amides Arg N[(CH 2 ) 5 ιMe](CH 2 ) 3 Ch Arg N(Me) n Bu Arg NH(CH 2 ) 3 CO Arg-NH 2 Arg NH(CH 2 ) 3 NH 2 Arg NH(CH 2 ) 4 CO Arg-NH 2 Arg NH(CH 2 ) 4 NH 2 Arg NH(CH 2 ) 5 CO Arg-NH 2 Arg NH(CH 2 ) 5 NH 2 Arg NH(CH 2 ) 6 CO Arg-NH 2 Arg NH(CH 2 ) 7 CO Arg-NH 2 Arg NH(CH 2 ) 7 CONH(CH 2 ) 3 Me Arg NH(CH 2 ) 7 NHAc Arg NH(CH 2 ) 7 NH 2 Arg NH(CH 2 ) 7 CONH 2 Arg NH(CH 2 ) 7 CO-Gly Gly Arg-NH 2 Arg NH(CH 2 ) 7 CO-Gly Arg-NH 2 Arg NH(CH 2 ) 7 CO-Gly Gly-Gly Arg-NH 2 Arg NpHex] 2 Arg NHCH 2 Ch Arg NHCH 2 Ch Arg NHCH 2 Ch Arg NHCH 2 Ch Arg NHCH 2 Ch Nar NHCH 2 Ch Arg NHCH 2 Ch Arg NHCH 2 Ch Arg NHCH 2 Ch Arg NHCH 2 Ch * Satisfactory amino acid analysis obtained H-DPro Phe Arg CH 3 H-DPro Phe Arg (CF 2 ) 2 CF 3 H-DPro Phe ArgCH 2 CH[ n Hex] 2 ** Molecular ion not detected TABLE 9 α-Ketoamides [M+HT Class H-DPro Phe DL Lys CONΓBU] 2 530 XI H-DPro Cha DL Arg CONPBu] 2 *** XI *** [M+H] + not available EXAMPLE I 5 Me-DPhe-Cha-Arg-H The synthesis of 5 was carried out according to Scheme I. Arabic numerals underlined e.g. 1 refer to structures in these schemes. Roman numerals in parentheses e.g. (i) refer to reaction steps. (i) Isobutyl chloroformate (10.2 mmol) was added to a solution of Boc-Arg(Z 2 )OH (9.23 mmol) and N-methylmorpholine (11.08 mmol) in dry THF (25 cm 3 ) at -20°C After 20 mins the solid was filtered off and the filtrate added to a solution of sodium borohydride (10.3 mmol) in water (10 cm 3 ) at 0°C After 3 hours 0.3 M KHSO 4 was added, the crude product extracted with EtOAc and purified by flash chromatography on silica with EtOAc - petrol (4:6). The alcohol 1 was isolated as a white solid (97%). (ii) The Boc group of i (4.75 mmol) was removed with sat. HCl Dioxan and the product acylated with Boc-Cha-ONSu (9.5 mmol) in CH 2 CI 2 (20 cm 3 ) at 0°C in the presence of N-methylmorpholine. After two hours the reaction was worked up using standard procedures and the crude product purified by flash chromatography on silica with EtOAc - petrol (4:6). The pure alcohol 2 was isolated as a colourless oil (90%). (iii) The Boc group of 2 (4.27 mmol) was removed with sat HCl Dioxan and the product reacted with Z(NMe)DPhe-OH (5.12 mmol) in the presence of HOBt (10.2 mmol), water soluble carbodiimide (6.1 mmol) and N-methylmorpholine in DMF (20 cm 3 ) at 0°C. After 18 hours the reaction was worked up using standard procedures and the product purified by flash chromatography on silica with EtOAc - petrol (1:1). The pure alconol 3 was isolated as a colourless oil (52%). (iv) The alcohol 3 (2.22 mmol) was dissolved in CH 2 Cl 2 /AcOH (30:1) and Dεss-Martin Periodinane (4.5 mmol) added. After 2\ hours at room temperature the reaction mixture was diluted with EtOAc and poured into a solution of sodium thiosulphate (32 mmol) and sat. NaHCO 3 . The crude product was purified by flash chromatography on silica with EtOAc-petrol (3:7). The pure aldehyde 4 was isolated as a colourless oil (75%). (v) The aldehyde 4 (1.65 mmol) was dissolved in MeOH/H 2 O/AcOH (90:9:1, 50 cm 3 ) and hyάrogenated over 5% Pd/C. The crude material was purified by mplc on *Vydac g (15-25 μ) using MeCN/H 2 O/TFA to give pure 5 (CH-851) as a white solid (780 mg). Tic, EtOAc-Py-AcOH-H 2 O (30:20:6:11), R F 0.66 on silica. After hydrolysis at 110°C/22 hrs with 6N HCI peptide content based on Cha was 40%. FAB mass spec [M+H] + = 473 (Calc. m /z = 472). EXAMPLE n ϋ H-DPro-Phe-Lys-CON π Bu 2 (see Scheme II) (i) TcbocONSu (14.8 mmol) was added to a solution of H-Lys(Z)-OMe. HCI (12.2 mmol) and triethylamine (14.8 mmol) in CH 2 C1 2 (50 cm 3 ). After 3 hours at room temperature the reaction was worked up using standard procedures and the product purified by flash chromatography on silica using EtOAc - petrol (7:13). The pure ester 6 was isolated as a colourless oil (100%). (ii) Diisobutylaluminium hydride (1.5 M solution in toluene, 50 mmol) was added to a solution of 6 (12.2 mmol) in dry toluene (100 cm 3 ) at -78°C over a period of 20 minutes. After a further 15 minutes methanol (10 cm 3 ) was added followed by a saturated solution of Rochelle's salt (100 cm 3 ). After 2\ hours the reaction was worked up using standard procedures and the product purified by flash chromatography on silica using EtOAc - petrol (3:7). The pure aldehyde 7 was isolated as a colourless oil (49%). * Trade name (iii) Potassium cyanide (18 mmol) and 1 M hydrochloric acid (30 cm 3 ) were added to a solution of 7 (5.98 mmol) in ethyl acetate (30 cm 3 ). After 18 hours at room temperature the reaction was worked up using standard procedures and the product purified by flash chromatography on silica using EtOAc - petrol (4:6). The pure cyanohydrin 8 was isolated as a colourless oil (88%). (iv) A 4 M solution of HCI in dioxan (50 cm 3 ) was added to a solution of 8 (5.28 mmol) in dry methanol (15 cm 3 ) at O°C. After 18 hours at room temperature an ice/water mixture (15 cm 3 ) was added. After 3 days at 4°C solid KHCO 3 was added. The reaction was worked up using standard procedures and the product purified by flash chromatography on silica using EtOAc - petrol (11:9). The pure ester 8b was isolated as a yellow oil (59%). (v) Activated zinc dust was added in small portions to a solution of 8b (3.1 mmol) in AcOH H O (9:1, 25 cm 3 ). After li hours at room temperature the zinc was filtered off, the filtrate evaporated in vacuo and the residue was taken up in EtOAc. This solution was washed with sat. NaHCO 3 , water, brine, dried (Na 2 SO 4 ) and evaporated in vacuo. The amine 9 was isolated as a colourless oil (85%). (vi) The amine 9 (2.63 mmol) was acylated with Boc-Phe-ONSu (3.04 mmol) in CH 2 C1 2 (30 cm 3 ) at 0°C in the presence of N-methyl morpholine. After 3 hours the reaction was worked using standard procedures and the crude product purified by flash chromatography on silica with EtOAc/Pet Ether (6:4). The pure ester JOa was isolated as a colourless oil (92%). (vii) The Boc group of iQa (2.41 mmol) was removed using sat. HCl/Dioxan and the product acylated with Boc-DPro-ONSu (2.92 mmol) in CH 2 C1 2 (30 cm 3 ) at 0°C in the presence of N-methyl-morpholine. After 3 hours the reaction was worked up using standard procedures and the product purified by flash chromatography on silica using EtOAc/Petrol (3:1). The pure ester 10b was isolated as a colourless oil (64%). (viii) Lithium hydroxide (1.6 mmol) and water (3 cm 3 ) were added to a solution of Ob (1.54 mmol) in THF (30 cm 3 ). After 4 hours at room temperature the THF was removed in vacuo, the pH of the residue adjusted to pH 4 with 1 M citric acid and extracted with CHCI 3 . The organic extracts were washed with brine, dried (Na 2 SO 4 ) and evaporated in vacuo. The pure acid jOc was isolated as a colourless oil (70%). (ix) Pentafluorophenol (1.3 mmol) and water soluble carbodiimide (1.3 mmol) were added to a solution of iOc (1.07 mmol) in CH 2 C1 2 (20 cm 3 ) at O°C. After 2 hours dibutylamine (2.1 mmol) was added to this solution at O°C and the pH adjusted to pH 9 with DIEA. After 18 hours at room temperature the reaction was worked up using standard procedures and the product purified by flash chromatography on silica using EtOAc/Petrol (7:3). The pure amide K)d was isolated as a colourless oil (48%). (x) Dess-Martin Periodinane (0.97 mmol) was added to a solution of lOd (0.52 mmol) in CH 2 Qz (100:1, 40 cm 3 ). After 2 hours at room temperature further Dess-Martin Periodinane (0.52 mmol) was added. After a further 3 hours the reaction mixture was diluted with EtOAc and poured into a solution of sodium thiosulphate (7.3 mmol) in water and sat NaHCO 3 were added. The crude product was purified by flash chromatography on silica with EtOAc/petrol (9:11). The pure keto amide JOe was isolated as a colourless oil (57%). (xi) he Boc group of iOe (0.24 mmol) was removed using sat. ..iCl/Dioxan. The resultant product was dissolved in AcOH/H 2 O (9:1) and hydrogenated over 5% Pd/C. The crude material was purified by mplc on *Vydac C 18 (15 - 25 μ) using MeCN/H 2 0/TFA to give ϋ (CH-1463 89.7 mg). Hplc, *Novapak C 18 , 4 μ (8 x 100 mm), linear gradient 20 -→ 80% 0.1% TFA/MeCN into 0.1% TFA/H 2 O over 25 min at 1.5 ml min "1 indicates th presence of two epimers D-Arg (40%) at 11.2 min and L-Arg (60%) at 12.6 min. After hydrolysis at 110°C/22 h with 6N HCI, amino acid analysis Phe 0.93, Pro 1.07. EXAMPLE m 17 H-DPro-Phe-Arg-CH 2 S(CH 2 ) 4 CH 3 (see Scheme m) (i) Boc-Arg(Z 2 >OH (46.1 mmol) was dissolved in dry THF (200 cm 3 ). N-methylmorpholine (50.85 mmol) and isobutyl chloroformate (50.73 mmol) were added at -20°C. After 20 min. this mixture vas added to a solution of diazomethane (0.1 mole) in Et 2 O at -5°C. After 2 hours the diazoketone . 12 was isolated as a yellow solid. (ii) The diazoketone i_2 (46.1 mmol) in dry THF was treated with HBr (69.15 mmol) in EtOAc at -20°C followed by addition of sat. NaHCO 3 after 45 mins. The crude product was extracted with EtOAc and crystallised from EtOH to give pure Boc-Arg(Z 2 )CH 2 Br, .13, (85%). (iii) 1-Pentanethiol (1.27 mmol) in dry DMF (5 cm 3 ) was treated with sodium hydride (1.4 mmol). After 30 mins Boc-Arg(Z 2 )CH 2 Br * 3_ (1.27 mmol) wa ; added-40°C for 20 mins and -5°C for 2\ hcjs. After addition of 0.3 M KHSO 4 and extraction of the crude product with EtOAc, flash chromatograpy on silica with EtOAc - petrol (15:85) yielded the pure thiometnylene compound .14 as a colourless oil (74%). * Trade name (iv) The Boc protecting group of i4 (0.94 mmol) was removed using sat. HCl/Dioxan and the resulting product was acylated with Boc-Phe-OPfp (1.13 mmol) in CH 2 C1 2 at 0°C in the presence of DIEA. The crude product was purified by flash chromatography on silica with EtOAc - petrol (3:7) yielding the pure thiomethylene analogue i5_ as a colourless oil (55%). (v) The Boc protecting group of 5 (0.52 mmol) was removed using sat. HCl/Dioxan. The resulting product was dissolved in DMF and treated with Boc-DPro-OH (0.63 mmol) in the presence of HOBt (1.05 mmol), water soluble carbodiimide (0.76 mmol) and N-methylmorpholine. After a standard work-up the crude material was purified by flash chromatography on silica with EtOAc - petrol (4:6) yielding the pure thiomethylene compound JL6 as a colourless oil (74%). (vi) The Boc protecting group of . 16 (0.38 mmol) was removed using sat. HCl/Dioxan. The resultant product was dissolved in AcOH/H 2 O (9:1) and hydrogenated over 5% Pd/C. The crude material was purified by mplc on *Vydac C 18 (15-25 μ), using MeCN/H 2 O/TFA to give pure 17 (CH-574, 41 mg). Hplc, *Novapak C 18 , 4μ (8 x 100 mm), Unear gradient 20 -→ 80% 0.1% TFA MeCN into 0.1% TFA H 2 O over 25 min at 1.5 ml min" 1 indicates the presence of two epimers D-Arg (<5%) at 9.8 min and L-Arg (>95%) at 11.1 min. After hydrolysis at 150°C/1.5 h with 6N HCI, amino acid analysis Phe, 0.80; Pro, 1.00. All analogues in Table 2 were synthesised by the described method. 61 was synthesised by the oxidation of 60 with meta- chloroperoxybenzoic acid. * Trade name EXAMPLE IV 23 H-DPro-Phe-Arg-CH 2 OCH 2 (CF 2 ) 3 CHCF 2 (see Scheme IV) (i) IH, IH, 5H-Octafluoro-l-pentanol (2.45 mmol) in dry DMF (8 cm 3 ) was treated with sodium hydride (1.83 mmol). After 30 mins the bromoketone J (1.65 mmol) was added at -40°C and left at this temperature for 30 mins and -5°C for 2\ hours. Addition of 0.3 M KHSO 4 and extraction with EtOAc gave the crude product which was purified by flash chromatography on silica using EtOAc - petrol (15:85). The pure fluoroether .18 was isolated as a colourless oil (69%). (ii) The fluoroether .18 (1.13 mmol) was dissolved in MeOH (40 cm 3 ), sodium borohydride (1.18 mmol) was added to this solution at 0°C. After 15 min 0.3 M KHSO 4 ^s added and the mixture extracted with EtOAc giving the pure compound .19 as a colourless oil (88%). (iii) The Boc protecting group of .19 (1.0 mmol) was removed with sat. HCl/Dioxan. The resulting product was dissolved in CH 2 CI 2 and acylated with Boc-Phe-OPfp (1.2 mmol) in the presence of DEEA at 0°C. After a standard work up the crude product was purified by flash chromatography on silica with EtOAc - petrol (3:7) yielding the pure product 20 as a colourless oil (55%). (iv) 20 (0.55 mmol) wr - deprotected with sat. HCl/Dioxan and acylated with Boc-DPro-OKp (1.63 mmol) in CH 2 C1 2 at 0°C in the presence of DIEA. After a standard work up the crude product was purified by flash chromatography on silica with EtOAc - petrol (4:6) yielding the pure product 2 as a colourless oil (48%). (v) 21 (0.24 mmol) was dissolved in O-^ WAcOH (30:1) and Dess-Martin Periodinane (0.48 mmol) was added. After 2 hours at room temperature the reaction mixture was diluted witii EtOAc and poured into a solution of sodium thiosuiphate (3.5 mmol) in water and sat. NaHCO 3 - The crude product was purified by flash chromatography on silica with EtoAc - petrol (7:13) yielding the pure fluoroether 22 as a colourless oil (64%). (vi) The fluoroether 22 (0.16 mmol) was deprotected and purified as described in Example in (vi). Pure 23 (CH-619) was isolated as a white solid (50.9 mg). Hplc, *Novapak C 18 4μ (8 x 100 mm), linear gradient 20 → 80% 0.1% TWMeCN into 0.1% TFA/H 2 O over 25 mins at 1.5 ml min *1 indicated a single product (TR = 11.5 min). After hydrolysis at 150°C 1.5 hr with 6N HCI, amino acid analysis Phe, 1.00; Pro, 1.2. All analogues in Table 3 were synthesised by the described method except 54 and 55 which were synthesised by methods outlined in Schemes XIII and XTV respectively. EXAMPLE V 3J. H-DPro-Phe-Arg-CH 2 N[(CH 2 ) 5 CH 3 ] 2 (see Scheme V) (i) Boc-Arg(Z2)OH (18.5 mmol) was dissolved in CH 2 C1 2 (50 cm 3 ). To this solution at 0°C was added trichloroethanol (20.35 mmol), water soluble carbodϋmide (22.2 mmol) and dimethylaminopyridine (0.93 mmol). After 3 hours the reaction was worked up using standard procedures giving the pure trichloroethyl derivative 24 as a colourless oil (100%). * Trade name (ii) 24 (18.1 mmol) was deprotected with sat. HCl Dioxan and acylated with Boc-Phe-ONSu (27.2 mmol) in CH 2 C1 2 at 0°C in the presence of N-methylmorpholine. After 3 hours the reaction mixture was worked up using standard procedures and the crude product was purified by flash chromatography on silica with EtOAc - petrol (2:8) yielding the pure product 25 as a white solid (97%). (iii) 25 (17.4 mmol) was deprotected with sat. HCl/Dioxan and acylated with Boc-DPro-ONSu (26.2 mmol) in CH 2 C1 2 at 0°C in the presence of N-methylmorpholine. After 3 hours the reaction was worked up using standard procedures and the crude product purified by flash chromatography on silica with EtOAc - petrol (35:65) giving the pure protected tripeptide 26 as a colourless oil (94%). (iv) Activated zinc powder was added to a solution of 26 (16.47 mmol) in glacial acetic acid. After 3 hours at room temperature the zinc was filtered off, the filtrate evaporated and the crude product purified by flash chromatography on silica with EtOAc - petrol - i.cOH (74:25:1) giving the pure tripeptide 27 as a white solid (91%). (v) The protected tripeptide 27 (15 mmol) was dissolved in dry THF (40 cm 3 ), N-methylmorpholine (18 mmol) and isobutyl- chloroformate (lc. mmol) were added at -20°C. After 20 mins the mixture was added to a solution of diazomethane (35 mmol) in Et 2 0 at -5°C. After 2i hours the diazoketone 28 was isolated as a yellow oil. (vi) The diazoketone 28 (15 mmol) in dry THF was treated with HBr (22.5 mmol) in EtOAc at -20°C followed by addition of sat. NaHCO 3 after 45 mins. The crude product was extracted with EtOAc and purified by flash chromatography on silica with EtOAc - petrol (1:1). The pure bromoketone 29 was isolated as a white solid (72%). (vii) Dihexyiamine (1.25 mmol) and NaHC0 3 (0.8 mmol) were added to a solution of bromoketone 29 (0.23 mmol) in dry THF (5 cm 3 ). After 18 hrs at room temperature 0.3 M KHS0 4 was added to the reaction mixture and the crude product was purified by flash chromatography on silica with EtOAc - petrol (35:65). The protected aminomethylene ketone 30 was isolated as a yellow oil (54%). (viii) The aminomethylene ketone 30 (0.12 mmol) was deprotected and purified as described in Example ffi (vi). Pure 3_i (CH-694) was isolated as a white solid (41 mg). Hplc, linear gradient 20 —» 80% 0.1% TFA/MeCN into 0.1% TFA/H 2 O over 25 mins at 1.5 ml min" 1 indicated the presence of two epimers D-Arg (50%) at 11.2 min and L-Arg (50%) at 12.5 mins. After hydrolysis at 110°C/22 hrs with 6N HCI, amino acid analysis Phe, 0.91; Pro, 1.09. All analogues in Table 4 were synthesised by the described method. The required amines were synthesised by standard synthetic methods such as reductive amination and the Curtius rearrangement. EXAMPLE VI 40 H-DPro-Phe-Arg≤Gly-Pro-NHEt (see Scheme VI) DL (i) H 2 C(CO 2 Tce) 2 (6.81 mmol) was treated with sodium hydride (5.67 mmol) in dry THF (30 cm 3 ). After 45 mins the bromoketone ϋ (4.52 mmol) was added at -5°C. After 2\ hours 0.3 M KHSO 4 was added, the crude product extracted with EtOAc and purified by flash chromatography on silica with EtOAc - petrol (2:8). The pure Boc-Arg(Z 2 )CH 2 CH(Cθ 2 Tce) 2 32 was isolated as a colourless oil (83%). (ii) Activated zinc was added to a solution of 32 (3.65 mmol) in glacial acetic acid. After 2\ hours at room temperature the zinc was filtered off, the filtrate evaporated and the diacid 33 isolated (iii) A solution of the diacid 33_ in toluene was heated at reflux for 45 mins. The solvent was evaporated and the crude product purified by flash chromatography on silica with EtOAc - petrol - AcOH (60:39:1). The Boc-ArgtZ^^Gly-OH 34 was isolated as a colourless oil (70% from 32). (iv) Trichloroethanol (2.82 mmol), water soluble carbodiimide (2.81 mmol) and dimethylaminopyridine (0.117 mmol) were added to a solution of 34 (2.34 mmol) in CH 2 C1 2 (50 cm 3 ) at 0°C. After 2 hours the reaction was worked up using standard procedures and the crude product purified by flash chromatography on silica with EtOAc - petrol (85:15). The trichloroethyl derivative 35 was isolated as a colourless oil (83%). (v) The Boc protecting group of 35 ( 1.85 mmol) was removed using sat. HCl/Dioxan and the resulting product acylated with Boc-Phe-OPfp (6.04 mmol) in CH 2 2 in the presence of DDΞA. After 2 hours the reaction was worked up using standard procedures and the crude product purified by flash chromatography on silica with EtOAc - petrol (2:8). The pure product 36 was isolated as a colourless oil (85%). (vi) The Boc protecting group of 36 (1.58 mmol) was removed using sat. HCl/Dioxan and the resulting product acylated with Boc-DPro-OPfp (5.12 mmol) in CH 2 C1 2 in the presence of DIEA. After 2 hours the reaction was worked up using standard procedures and the crude product purified by flash chromatography on silica with EtOAc - petrol (35:65). The pure product 32 was isolated as a colourless oil (79%). (vii) Activated zinc dust was added to a solution of 37 (1.13 mmol) in glacial acetic acid. After 2\ hours at room temperature the zinc was filtered off, the filtrate evaporated and the crude product purified by flash chromatography on silica with EtOAc - petrol - AcOH (70:29:1). The product 38 was isolated as a colourless oil (76%). (viii) The protected keto isostere 38 (0.26 mmol) was convened to its Pfp ester by treatment with Pfp-OH (0.29 mmol) and water soluble carbodiimide (0.31 mmol) in CH 2 C1 2 (8 cm 3 ) at 0°C for Ik hours. This Pfp ester was coupled at 0°C to H-Pro-NHEt . HCI salt (0.78 mmol) in the presence of DIEA. After 18 hours the reaction was worked up using standard procedures and the product purified by flash chromatography on silica with CHCl 3 -MeOH-AcOH (97:2:1). The product 39 was isolated as a colourless oil (91%). (ix) The protected keto isostere containing analogue 39 (0.23 mmol) was deprotected as described in Example HI (vi). Pure 40 (CH-595) was isolated as a white solid (40 mg). Hplc, linear gradient 10 -→ 50% 0.1% TFA/MeCN into 0.1% TFA/H 2 O over 25 mins at 1.5 ml min "1 indicated the presence of two epimers D-Arg (46%) at 10.1 min and L-Arg (54%) at 11.7 min. After hydrolysis at 150°C/1.5 hrs with 6N HCI amino acid analysis Phe, 0.91; Pro, 1.09. All analogues in Table 5 were synthesised by the described method. EXAMPLE VII 45 H-DPro-Phe-Arg-Chg-NH 2 (see Scheme VH) (i) Boc-Phg-OH (19.9 mmol) was dissolved in AcOH/H 2 0 (9:1, 100 cm 3 ) and hydrogenated over Rh/C at 60 p.s.i. for 3 days. The catalyst was filtered off and the solvent removed to give Boc-Chg-OH 4_i (100%). (ii) Water soluble carbodiimide (4.1 mmol) and HOBt (4.3 mmol) were added to a solution of 4J_ (3.9 mmol) in CH 2 C1 2 /DMF (2:1, 60 cm 3 ) at room temperature. After 30 mins 35% ammonia solution (0.8 cm 3 ) was added. After a further 3 hours at room temperature the reaction was worked up using standard procedures and the product recrystallised from EtOH to give the pure amide 42 as a white solid (60%). j (iii) The Boc group of 42 (0.77 mmol) was removed with sat. HCl Dioxan to give the amide 43 as a white solid (100%). (iv) The protected tripeptide 27 (0.38 mmol) was dissolved in DMF (5 cm 3 ). 43 (0.76 mmol), HOBt (0.76 mmol) water soluble carbodiimide (0.46 mmol), and N-methylmorpholine were added at 0°C. After 18 hours at room temperature the reaction was worked up using standard procedures and the product purified by flash chromatography on silica with CHC- 3 /MeOH (99:1). The pure protected tetrapeptide 44 was isolated as a white solid (69%). (v) The protected tetrapeptide 44 (0.27 mmol) was deprotected and purified as described in Example IH (vi). Pure 45 (CH-640) was isolated as a white solid (58 mg). Hplc, linear gradient 10 — * 50% 0.1% TFA/MeCN into 0.1% TFA H 2 O over 25 min at 1.5 ml min" 1 , single peak detected at 14.2 min. After hydrolysis at 110°C/22 hrs with 6N HCI, amino acid analysis, Arg, 0.96; Phe, 1.00; Pro, 0.95. All analogues in Table 6 were synthesised by the described method or by other standard peptide coupling methodology. (M. Bodansky & A. Bodansky, The Practice of Peptide Synthesis, Springer- Verlag, 1984) EXAMPLE Vm 47 H-DPro-Phe-Arg-N[(CH 2 ) 5 CH 3 ](CH 2 ) 3 Ch (see Scheme VHI) i) The protected tripeptide 27 (0.32 mmol) was dissolved in DMF (5 cm 3 ), HN[(CH 2 ) 5 CH 3 ](CH 2 ) 3 Ch (0.96 mmol), HOBt (0.64 mmol), water soluble carbodiimide (0.38 mmol) and N-methylmorpholine were added at O°C. After 18 hours at room temperature the reaction was worked up using standard procedures and the product purified by flash chromatography on silica using EtOAc Hexane (4:6). The amide 46 was isolated as a colourless oil (32%). (ii) The protected tripeptide amide 46 (0.1 mmol) was deprotected and purified as described in Example IE (vi). Pure 47 (CH-985) was isolated as a white solid (17 mg). Hplc, linear gradient 40 — * 90% 0.1% TFA/MeCN into 0.1% TFA/H 2 O over 25 min at 1.5 ml min "1 , single peak detected at 10.7 min. After hydrolysis at 110°C for 22 hrs with 6N HCI, amino acid analysis Phe, 1.01; Pro, 0.99. All analogues in Table 7 were synthesised by the described method. The required tripeptide presursors were synthesised in a similar manner to 27 or by standard peptide coupling methodology. (M. Bodansky & A. Bodansky, The Practice of Peptide Synthesis, Springer- Verlag, 1984.) Required amines were either commercially available or synthesised via the Curtius rearrangement. n Bu-DPro-OH for i8_l was synthesised by reductive amiπation. Scheme 1 (Synthesis of compound 5) Z (i) -BuOCOCl NMM/THF i ", ^2 Boc-Arg-OH Boc-Arg-OH NaBHi/HO 1 (ii) ~-ι (iii) HCl/Dioxan Z, ! Z(NMe)DPhe-Cha-Arg ≤ OH .4 Boc-Cha-Arg ^OH Z(NMe)DPhe-OH HOBt/wscd Z(NMe Me-DPhe-Cha-Arg-H MeOH H-,0/AcOH 4 Scheme II (Synthesis of compound U Z (i) TcbocONSu Z H-Lvs-OMe . HCI Tcboc-Lys-OMe E NVCH l, (ii) DIBAlJToluene Z liii) KCN Z Tcboc-Lys Q HCN Tcboc-Lys-H (iv Z I (v) Zn/AcOH Tcboc-Lys^CO 2 Me H-Lys Q HCOiMe 8b Boc-DPro-Phe- (ix) PfpOH/wscd CH 2 α 2 Z Boc-DPro-Phe-Ly S Q HCO 2 H +■ Boc-DPro-Phe-Lys QH CON n Bu, 10c n Bu,NH/DIEA lOd (x) Dess Martin Periodinane (xi) HCl/Dioxan H-DPro-Phe-Lys-CON^B^ Boc-DPro-Phe-Lys-CON n Bu 1 H 2 , Pd C 11 . AcOH/H 2 O lOe Scheme III (Synthesis of compound 17) (i) -BuOCOCI/NMM ΗF ^2 Boc-Arg-OH Boc-Arg-CHN 2 (ii) CH,N, 12 , I ~ Boc-Arg-S(CH 2 ) 4 CH 3 CH-Br 14 DMF 13 7 ,r -- (v) HCl Dioxan Z-. Boc-Phe-Arg-CH 2 S(CH 2 ) 4 CH 3 *- Boc-DPro-Phe-Arg-CH 2 S(CH 2 ) 4 CH ? H-DPro-Phe-Arg-CH 2 S(CH 2 ) 4 CH 3 17 Scheme IV (Synthesis of compound 23) Z, (i) CHF 2 (CF 2 ) 3 CH 2 O"Na + Z, i " I Boc-Arg-CH 2 Br >Boc-Arg-CH 2 OCH 2 (CF 2 ) 3 CHF 2 DMF 13 18 (ii) N BH 4 /MeOH (iii) HCl Dioxan Boc-Phe-Arg^iCH 2 OCH 2 (CF 2 ) 3 CHF 2 -*- Boc-ArgQHCH 2 OCH 2 (CF ; ) 3 CHF ; (iv) Boc-DPro-Phe-Arg°£CH 2 OCH 2 (CF 2 ) 3 CHF 2 (v) Z, i ~ Boc-DPro-Phe-Arg-CH 2 OCH 2 (CF 2 ) 3 CHF 2 (vi) H-DPro-Phe-Ar -CH2θCH 2 (CF 2 3 CHF 2 23 Scheme V (Synthesis of compound 3J_) Z, (i) Tce-OH/wscd Z i ~ i - Boc-Arε-OH Boc-Ars-OTce DMAP/CHiCL, 24 (ii) HCl/Dioxan Boc-Phe-ONSu CH-.QVNMM (iii) HCl/Dioxan Z, Boc-DPro-Phe-Arε-OTcε Boc-Phe-Arg-OTce (iv) Zl Boc-DPro- -Arg-CHN : (vi) HBr/EtOAc THF (vii) HN[(CH 2 ) 5 CH 3 ] 2 Z, Boc-DPro-Phe-Arg-CH 2 N[(CH 2 ) 5 CH 3 ] 2 Boc-DPro-Phe-Arg-CH 2 Br NaHCO-, HF 30 29 (viii) HCl/Dioxan H 2 , Pd/C, AcOH, H 2 O H-DPro-Phe-Arg-CH 2 N[(CH 2 ) 5 CH 3 ] 2 31 Scheme VI (Synthesis of compound 40) Z, (i) Na + -CH(CO-,Tce), Boc-Arg-CH 2 Br Boc-Arg-CH 2 CH(CO 2 Tce) 2 13 (ii) Zn/AcOH (iii) Toluene / Δ Boc-Arg^Gly-OH Boc-Arg-CH-.CH(CO H , 34 -^ Boc-Phe-OPfp 35 36 DIEA/CH-,C1, (vi) HCl/Dioxan Boc-DPro-OPfD CH 2 C1 2 /DIEA ' (vii) Zn/AcOH Boc-DPro-Phe-ArgϋGly-OH ■* -* *■ Boc-DPro-Phe-Arc^Glv-OTce 38 H 2 , Pd/C 39 AcOH/H 2 O 40 I D Scheme Vn (Synthesis of compound 45) (i) H 2 , Rh/C Boc-Phg-OH Boc-Chg-OH AcOH/H 2 O/60 psi 41 (iii) HCl/Dioxan H-Chg-NH 2 HCI Boc-Chg-NH 2 43 42 (iv) 27 HCl/Dioxan Boc-DPro-Phe-Arg-Chg-NH 2 H-DPro-Phe-Arg-Chg-NH 2 H 2 , Pd/C 44 45 AcOH/H 2 O Scheme VIII (Synthesis of compound 47) ^ (i) HN[(CH 2 ) 5 CH 3 ] (CH 2 )f h ^ Boc-DPro-Phe-Arg-OH *- Boc-DPro-Phe-Arg-N[(CH 2 ) 5 CH 3 ](CH 2 ) 3 Ch 27 HOBt/wscd/DMF 6 (ii) H-DPro-Phe-Arg-N[(CH 2 ) 5 CH 3 ](CH 2 ) 3 Ch 47 Scheme IX (Svnthesis of compound 48^ Z- Boc-ArgSOH 1 Z, CF 3 CF 2 CF 2 I Z-, I 3oc-Arg-H Boc-Arg2HCF 2 CF 2 CF, Zn/THF Ultrasound 3oc-DPro-Phs- Boc-DPro- H-DPro-Phe-Arg-CF 2 CF 2 CF 3 4 δ Scheme X (Synthesis of compound 49) Zn/AcOH Boc-DPro-Phe-Arg-CH 2 Br Boc-DPro-Phe-Arg-CH 3 29 H-DPro-Phe-Arg-CH 3 49 Scheme XI (Syntnesis of compound 50) 3oc-Arg-CH 2 Br Boc- HCl Dioxan oc-Arg Q H- Q Sco,Me Boc-Cha-Arg2I2DMs C o 2Me Boc-Cha-ONSu CH 2 C1 2 /NMM HCl/Dioxan Boc-DPro-ONSu NMM/CH 2 C1 2 H-DPro-Cha- Arg-CO 2 N n B u 2 Scheme XII (Synthesis of compound 53) • BuOCOClNMMTHF Boc-Orn-OH Boc-Orn-OH NaBH H 2 O H 2 Pd/C AcOH/H,0 :ONSu ΔTo LiBr/Acetone n Hex 2 CHCH 2 Br "Hex-.CHCHOMs 52 (cont.) Scheme XII (cont.) M /THF Bzl, Z π Hεx 2 CHCH 2 Br Boc-Om^G^O-PHex, 51 52 HCl/Dioxan Boc-Phe-ONSu CH,C1 2 /NMM Bzl. Z HCl Dioxan Bzl, Z 3oc-DPro-rhe-Crn^CH,CH T1 Hcx-, Boc-Phe-OmaHCH,CH n Hcx-. Boc-DPro-ONSu/NMM Dess Martin Periodinane CH 2 Cl 2 AcOH Z> HCl/Dioxan H-DPro-Phe-Arg-CH 2 CH n Hcx 2 Boc-DPro-Phe-Arg-CH 2 CH n Hex Scheme Xm (Synthesis of compound 54) Boc-DPro-Phe- COPh Boc-DPro-Phe-Arg-CH 2 O(CH 2 ) 5 CH 3 -* Boc-DPro-Phe-Arg-CH 2 OH CH 2 C1 2 H-DPro-Phe-Arg-CH 2 O(CH 2 ) 5 CH 3 54 Scheme XIV (Synthesis of compound 55) PhOH/KF > Boc-DPro-Phe-Arg-CH 2 Br ► Boc-DPro-Phe-Arg-CH 2 OPh DMF 29 HCl/Dioxan H 2 Pd/C AcOH/H 2 O H-DPro-Phe-Arg-CH 2 OPh 55 ABBREVIATIONS USED Abn 3-Azabicyclo[3.2.2]-nonane Ac Acetyi AcOH Acetic acid Ada Adamantylalanine Aha 2-Aminohexanoic acid (Norieucine ) Boc tert-B utyloxycarbony 1 Bu Butyl Ch Cyclohexyl Cha Cyclohexylalanine Chg Cyclohexylglycine Cpr Cyclopropyl DIEA Diisopropylethylamine DMAP 4-Dimethylamino-pyridine DMF Dime ylformamide EtOAc Ethyl acetate FAB . Fast Atom Bombardment 4-Fph 4-Fluorophenylalanine Hch Homocyclohexylalanine HOBt 1 -Hydroxybenzoniazole hplc high performance liquid chromatography Hyp 4-Hydroxyproline 'Hyp trans-4-Hydroxyproliπe BIOLOGICAL ACTIVITY; -MEDICAL USE Compounds were tested in vi ro for the following activities using standard procedures: (a) Inhibition of human tissue kallikrein, plasma kallikrein and mast cell tryptase hydrolysing the chromogenic substrates S-2266, S-2302 and S-2266 respectively (method is adapted from that of Johansen, H.T. , et al., Int. J. Tiss. Reac, 1986, a, 185-192). A series of measurements were carried out using a number of different; inhibitor concentrations and at least two different substrate concentrations. The inhibitory constant Ki was determined graphically, using a Dixon plor (M. Dixon , Biochem. J. , 1953, 5_5, 170). (b) Inhibition of kinin release from low and high molecular weight kininogens by tissue and plasma kallikrein respectively. A series of measurements were carried out using two substrate concentrations. The activity is calculated as the amount of kinen released per minute, this being determined by radioimmunoassay using polyclonal antibodies. The inhibitory constant Ki was determined graphically using a Dixon plot. All the examples in Tables 1 - 9 have Ki values in the K Q rraannggee 1100 33 -- 1100 MM aa<gainst one or all of the enzymes in the chromogenic assay, In vivo activity has been tested in well-established pharmacology models of asthma based on the sensitised guinea pig. A selection of these inhibitors representing the different chemical types proved to be highly effective in blocking both the acute phase response and the late phase reaction, their efficacy being comparable or superior to those of the topical steroids and B 2 -agonist currently used in asthma therapy. When the compounds of the present invention are used as a medicine, there are no critical limitations to the administration methods. The present enzyme inhibitor can be formulated by any conventional method in pharmaceutics. For example, the present enzyme inhibitor may be applied in any conventional manner including intravenous injection, intramuscular injection, instillation, oral administration, respiratory inhalation, instillation, rhinenchyεis, and external skin treatment. Although there are no critical limitations to the administration dosage, the suitable dosage is 1 to 1000 mg/day/person.

Claims: C L A I M S 1. Kininogenase inhibitors, optimally not exceeding the size of a hexapeptide, represented by:- 1 R- ~ I A - 3 - N I R wherein A and B = amino acyl (including amino acyl analogue) the same or different forming a dipeptide group the amino acid of A carrying a terminal group and being any amino or imino-acid residue (but preferably of D-configuration) and of B being a lipophilic amino-acid residue of D- or L- configuration but not proline or a proline analogue, or a conformational analogue of said dipeptide group wherein the peptide link is replaced by -CH 2 -NH- ('reduced'), -CH(OH)-CH 2 - ( 'hydroxy') , -CO-CH 2 - ( 'keto' ) , -CH 2 -CH 2 - ('hydrocarbon') or other conformational mimic of the peptide bond and in:- the side chain R 1 is that of a basic amino acid or amino acid analogue (preferably of L- configuration) and R is H or lower alkyl (C-^ - C 4 ) or C α or the peptide link comprising -N(R)- is replaced leading to a conformational mimic as above = a binding enhancing or carbonyl activating group preferably selected from H (when A or B must be cyclohexylalanine, preferably D if ar A or L if at B) or alkyl (C-^ - C 20 ) or fluoroalkyl (C 2 - C 12 ); substituted oxymethylene; thiomethylene; sulphoxy- methylene; sulphonylmethylene; aminomethylene; hydrazino-methylene; -CH 2 -Het (where Het = a substituted or unsubstituted heterocycle) ; substituted amino (but when the resulting compound is a secondary alkyla ide B must not be phenylalanine) ; an amino-acid group or its ester or amide; a carboxylic secondary amide or primary amide, when B must be cyclohexyl¬ alanine or adamantylalanine or other bulky lipophilic, non-aromatic amino-acid (not Ala Leu lie Val Nva Met Nle Phe Tyr Trp Nal (1)) ; tertiary-carboxamide; carboxy-alkyl group or its ester or amide or amino acyl derivative. 2. Compounds according to claim 1 wherein A is selected from imino-acids, (e.g. D-proline or an analogue of proline e.g. pipecolinic acid, azetidine carboxylic acid); lipophilic amino acids (e.g. DPhe, DCha, DChg) ; strongly basic amino acids (e.g. D-Arg or a homologue or analogue of Arg, e.g. amidino- or guanidinophenylalanine) ; or N-alkyl or C α -alkyl (including benzyl) derivatives thereof. 3. Compounds according to claim 1 or 2 wherein B is selected from L-Phe, L-Cha, L-αNal, L-Tal, L-(4F)Phe L-(NMe)Phe or other substituted phenyialanines; or N-alkyl or C 2 -alkyl (including benzyl) derivatives thereof. 4. Compounds according to claim 1, 2, or 3 wherein R is selected from 3-guanidinopropyl or other guanidinoalkyl group, (or an amidinoalkyl or aminoalkyl group), also para- or meta substituted guanidino or amidino-benzyl or protected forms of the above; optionally basic nitrogens are alkylated (Me, Et or other) . 5. Compounds according to any preceding claim wherein subject to the provisos in regard to the nature of Y expressed in claim 1, selection for Y is from:- Y = H or alkyl including fluoroalkyl Y = -CH 2 Q where Q = -OR 2 or -SR 2 or -SOR 2 or -SO-,R 2 or wherein R , R 4 and ^ are as below o κ - H CHR ό CO-D) wherein R 4 . R 5 and R 6 are as below Y = amino acyl or a group forming a substituted amide or hydrazide Y = a group forming an -keto amide - COR 9 or -CO-D^ or and in which further:- R 2 = alkyl or substituted alkyl including aryl or aryl alkyl or -CH 2 R 3 where R 3 = fluoroalkyl R 4 and R 5 the same or different but not both hydrogen = H or C 1 -C 20 alkyl (which may be further substituted) , acyl or alkyl sulphonyl -DJ is a heterocyclic ring (D = nitrogen or carbon in Group IV and N in Groups VI and X) optionally unsaturated and optionally with further hetero atoms and substituents R 6 = hydrogen, alkyl, hydroxyalkyl, aminoalkyl, alkylaminocarbonyl R = -NI- as such or alkylated, or ammo acyl. 6. Compounds according to any claim 1 to 4 wherein subject to the provisos in regard to the nature of Y expressed in claim 1, selection for Y is from:- Group I Y = H; alkyl including branched alkyl (Cτ_-C 20 ); aryl alkyl; or cycloalkyl (Cι_-C 20 ); perfluoroalkyl or partially fluorinated alkyl (C 2 -C 12 ); [e.g. Y = Me; -CH(CH CH-,CH-,CH-,)-W -CH(CH CH-,CH-,CH-,)CH^-cvclohexvl; —CH C j . Group II Y * = -CH 2 QR 2 where = O, S, SO, S0 2 , NH and where R- 4 = Alkyl, branched alkyl or acyl (C 1 -C 12 ); or cycloalkyl (Cι-C 20 ); or aryl or aryl alkyl; or -CH 2 -R J where R 3 = perfluoroalkyl or partially fluorinated alkyl, branched or not (C- j _-C 12 ). Group III R 4 Y = - CH- > N \ - 3 R 4 , R 3 the same or different = alkyl, branched alkyl, cycloalkyl, acyl, alkylsulphonyl, carboxyalkyl (the carboxyl group may be further derivatized to form an ester or amide with an amino-acid or dipeptide), carbamoyl, sulpha oyl, N-dialkylamino-, arylalkyl, haloalkyl including fluoroalkyl, cyanoalkyl, alkoxyalkyl, hydroxyalkyl, mercaptoalkyl, a inoalkyl and derivatives thereof e.g. esters, amides and thioesters; or one of R * * or R = hydrogen Group IV where D = nitrogen or carbon and -15 * ) is a saturated or unsaturated heterocyclic ring or a bicyclic ring system, each is 5 - 8 membered, where there may be other hetero- atoms (N, S, 0) and carbons or nitrogens may optionally be substituted by alkyl, branched alkyl, cycloalkyl, carboxyalkyl, carboxy (attached to carbon), amino, alkoxy, alkoxymethyl or (carbon) as carbonyl or other groups beneficial for interaction with the enzyme. Grouo V Y = -CK 2 CH(R 6 )CONR R 5 R 4 , R 3 as defined in Group III. R 6 = hydrogen lower alkyl, branched alkyl, cycloalkyl, hydroxyalkyl, amino-alkyl, alkylaminocarbonyl. Group VI R 6 as defined in Group V and - D) as defined in Group IV (but with D = N) Group VII Y = an amino-acid residue or any amide (secondary or tertiary) or ester of that residue, L or D configuration. Preferred residues are of lipophilic amino-acids e.g. norleucine, cyclohexylalanine, ho ocyclohexylalanine , cyclohexylglycine , ter"t: butylglycine . Group ->.' R 7 = H (when however B is not phenylalanine unless R 1 - i carboxylalkyl or derivatized carboxylalkyl); or alkyl, branched alkyl (C 1 - C 12 ) , cycloalkyl (C-,-C 20 ) carbσxyalkyl or bis ( carboxyl ) alkyl , which may b derivatized at the carboxyl group to form an amide e.g. with an amino-acid (preferred is arginine) or substituted amine; N'-dialkylamino; N'-alkylamino-; R 8 = R 7 the same or different but excluding H. Group IX Y = -CO-R y but only if B is a bulky non-aromati lipophilic amino acid or its N? alkyl (C ? - C 4 ) derivative (e.g. cyclohexylalanine but excluding Ala, Leu, lie, Val, Nva, Met, Nle, Phe, Tyr, Trp, Nal(l) an their N-methyl derivatives) where R 9 = NH ' N'-alkylamino (where the alkyl groups include branche and/or cycloalkyl); an amino-acid residue. Group X Y = -CO-D) ■ D) as defined in Group IV (but D - N) Group XI Y = -CO-NR"*R- R 4 ,R 5 as defined in Group III, but not H. 7. Any one of the compounds specifically listed in Tables 1 to 9 herein. 8. A method of treatment (including prophylactic treatment) of an inflammatory or other condition as set out in the indications (1) to (6) herein, particularly an allergic inflammatory condition, wherein an effective amount of a peptide or peptide-analogue kininogenase inhibitor is administered topically or systemically to a patient suffering from or at risk of the condition, the peptide or peptide analogue used being optimally of hexapeptide or smaller size. 9. A method of treatment of the allergic inflammatory phase of asthma, wherein an effective amount of a kininogenase inhibitor e.g. a mast cell tryptase inhibitor is administered topically or systemically to a patient suffering from or at risk of the condition. 10. A method of preparation of a medicament for the topical or systemic treatment (including prophylactic treatment) of conditions as in claim 8 particularly for allergic inflammatory conditions and especially for asthma as in claim 9, wherein a kininogenase inhibitor is associated in effective amounts with a pharmaceutically acceptable diluent or carrier to constitute said medicament. 11. A method of treatment or of preparation of a medicament as above wherein the inhibitor is a compound as claimed in any of claims 1 to 7.

Inventors: EVANS DAVID MICHAEL, JONES DAVID MICHAEL, SZELKE MICHAEL, EVANS, DAVID, MICHAEL, JONES, DAVID, MICHAEL, SZELKE, MICHAEL
Applicants: EVANS DAVID MICHAEL, FERRING PEPTIDE RESEARCH PARTN, JONES DAVID MICHAEL, SZELKE MICHAEL, EVANS, DAVID, MICHAEL, FERRING PEPTIDE RESEARCH PARTNERSHIP KB, JONES, DAVID, MICHAEL, SZELKE, MICHAEL
Application year: 1991
Main classification: A61K37
IPC codes: A61K38, A61P9, A61P25, A61P29, A61P37, C07K5, C07K14