MC2221A1 - ACETIC ACID DERIVATIVES - Google Patents

Abstract

Acetic acid derivatives of formula H2N(NH)C-X-Y-CO-Z-CH(Q1)COOQ2 (I), their hydrates, solvates and salts. In (I), Q1 = H, CH3 or phenyl. Q2 = H, phenyl-alkyl or physiologically-cleavable alkyl. X = 1,4-phenylene, 2,5- or 3,6-pyridylene or 1,4-piperidylene, which is bound to Y via the 4-C atom. Y = -(CH2)n-CONH-CH(Q3)-(CH2)m- (where n = 0-2 and m = 1-3); -CONH-CH2-CH(Q4)-, -(CH2)2-NHCO-CH2-, -NHCO-(CH2)3-, or a group of formula (a), (b) or (c). In the formulae, Q3 = H, CH3, phenyl, COOH, -COO-alkyl, -CONH-(CH2)2-COOH or -CONH-(CH2)2-COO-alkyl. Q4 = H, CH3 or phenyl. Z = 1,4-piperazinyl, 1,4-piperidinyl (which is bound to the CO group via the 1-N atom) or a group of formula -NH-CH(R1)- or -NH-CH(COR2). R1 = H, CH3, phenyl or COO-alkyl. R2 = an alpha-amino carboxylic acid linked via the amino group (or its ester or amide) or a group of formula -NH-(CH2)2-Ar, -CO2R2, an optionally mono- or dialkylated -CONH2 group, or a pyrrolidinoyl or piperidinoyl. Ar = phenyl optionally substituted by alkyl, alkoxy, -COOH, -CO-alkyl, -O(CH2)p-COOH (p = 1-4), O(CH2)p-COO-alkyl, -CONH2, -CONH-alkyl, -CO(alkyl)2, pyrrolidinoyl or piperidinoyl.

Description

acetic acid derivatives

The present invention relates to novel derivatives of acetic acid, a process for their preparation, pharmaceutical preparations containing such compounds, and the application of these compounds to the preparation of pharmaceutical preparations;

The invention relates in particular to acetic acid derivatives of the formula:

h2n(nh)c-x-y-co-z-ch(q1)cooq2

represents a hydrogen, methyl or phenyl,

represents a hydrogen, lower phenyl-alkoyl or a lower alkyl separable under physiological conditions, represents a 1,4-phenylene, 2,5- or 3,6-pyridylene or a bonded 1,4-piperidinylene

Q

2

X

2

via a carbon atom in position 4 of group Y,

represents a formula group

-(CH2)0-2"CONHCH(q3)(CH2)1-3'

(ch2)2nhcoch2-

■Çnt)o.ï-c o

\\

Or

(Y1)

2

■conhchjch(q4)- (y }

(y3)

4

-nhco(ch2)3- (y >

(y5)

(y6)

3

Q represents hydrogen, methyl, phenyl,

-COOH, -COO-lower alkyl, -CONH(CH2)2~ COOH or -CONH(CH2)2-COO-lower alkyl, Q4 represents a hydrogen, methyl or phenyl,

Z represents a 1,4-piperazinylene group,

a 1,4-piperidinylene group linked via nitrogen atoms in position 1 of the CO group or a group of formula

-NHCHCR1)- 2

or -NHCH(COR)_

represents a lower hydrogen, methyl, phenyl or -COO-alkoyl,

2

R represents the radical of an α-amino acid

carboxylic acid linked via the amino group, or one of its esters or amides, or a group of formula -NHCH2CH2-Ar, or -CO-R^ represents a carbamoyl group possibly mono- or disubstituted by a lower alkyl group or a pyrrolidinoyl or piperidinoyl group, Ar represents a phenyl or a substituted phenyl

by a lower alkyl, lower alkoxy, -COOH, -COO-lower alkyl,

-0(CH2)1_4-COOH, 0-CH2)1_4-COO-lower alkyl, -CONH2, -CONH-lower alkyl, -CON(lower alkyl)2,

pyrrolidinyl or piperidinoyl,

as well as their physiologically acceptable hydrates or solvation products and salts.

In the context of the invention, the abbreviations have the following meanings:

tBu t-butyl, Boc t-butoxycarbonyl, Z benzyloxycarbonyl Val L-valyl, Phe L-phenylalanyl, Ser L-seryl, Gly glycyl, Ala L-alanyl, Asp L-a-aspartyle, Leu L-leucine, Tyr L-tyrosine.

The term "lower" refers to groups having from 1 to 6, preferably from 1 to 4, carbon atoms. Examples of lower alkyl groups include methyl, ethyl, propyl, isopropyl, and n-, s-, or t-butyl. Primary and secondary lower alkyl groups are examples of lower alkyl groups that can be separated under physiological conditions.

Examples of α-amino-carboxylic acid esters linked via the amino group include Val, Phe, Ser, Leu, Tyr and their corresponding lower alkyl- or lower phenyl-alkyl esters, amides, and mono- or di-lower alkyl amides.

Compounds of formula I can be solvated, in particular hydrated. Hydration can occur during the preparation process or appear gradually as a result of the hygroscopic properties of a compound of formula I that is initially anhydrous.

Examples of physiologically acceptable salts of compounds of formula I include salts with physiologically acceptable mineral acids, such as hydrochloric acid, sulfuric acid, or phosphoric acid, or with organic acids, such as methanesulfonic acid.

Acetic acid, trifluoracetic acid, citric acid, fumaric acid, succinic acid, or salicylic acid. Compounds of formula I can also form salts with physiologically acceptable bases. Examples of such salts include alkali metal salts, alkaline earth metal salts, ammonium salts, and alkali-ammonium salts, such as Na-, K-, Ca-, or tetramethylammonium salts. Compounds of formula I contain an amidino group and can therefore exist as hermaphroditic ions.

One or more asymmetric carbon atoms may exist as enantiomers, diastereomers or mixtures thereof, e.g. as racemates.

The compounds of formula I, which contain

The preferred compounds of formula I are those of formula

20

H2N(HN)C-X-Ya-CONHCH(R11)CH_COOQ21

M

I-A

25 where X

represents a 1,4-phenylene or a 1,4-piperidinylene linked via the carbon atom at position 4 to the Ya group,

30 Ya represents a formula group

-<CH2)o-rCONHCH<Qa)(CH2)i-2

6

-cch2)2nhcoch2- (y3j

-nhco(ch2)3- (y4}

or CV51)

Qa R11

R22

a group

-ch2conh(ch2)2-,

-conhch(c6h5)ch2-,

-conhch(conhch2ch2cooh)ch2-.

Or

-conhch(cooh)ch2-

-conhch(ch3)ch2-.

represents a hydrogen or a phenyl group,

2

represents a hydrogen or -CO-R, represents the radical of an α-amino-carboxylic acid linked via the amino group or one of its esters or amides, and represents a hydrogen or lower alkyl separable under physiological conditions.

In formula I, Y is preferably of formula Y"*", in particular -conh(ch )2_4-, ;7 ;In formula I Z is preferably a group of formula ;-nhch2-, -nhchcch3)-, -nhch(c6h5)-, ;-NHCH(COO-Isobutyl)-, -NHCH(CO-Val)-. -NHCH(CO-Phe)-. -NHCH(CO-TyC)-, -NHCH(CO-Ser-OC2H5)-. -NHCH(CO-Leu-O-Isopropyl)-, ;-NHCH(CONHCH2CH2-C6H4-OCH3)-. -NHCH(CONHCH2CH2-C6H4-COOH)-, ;-NHCH(CONHCH2CH2-C6H4-OCH2COOH)-, ;-NHCH(CONH2)- -NHCH(Pyrrolidinoyl)- ;As particularly preferred compounds of formula I, the following should be mentioned; 5 N-[N-[N- (p-amidinobenzoyl)-3-alanyl]-L- ;a-aspartyl]-3-phenyl-L-alanine, ; 10: a-aspartyl]-3-(p-hydroxyphenyl)-L-alanine, ;N-[N-[5-(p-amidinobenzamido)valeryl]-L-a-aspartyl]-3-phenyl-L-alanine, ;N-[5-(p-amidinobenzamido)valeryl]-L-a-aspartic acid, isobutyl ester, 20 N-[N-[N-(p-amidinobenzoyl)-3-alanyl]-L- ;a-aspartyl]-L-serinethyl ester and ;N-[N-[[(R)-l-(p-amidinobenzoyl)-3-pyrrolidinyl]-carbonyl]-L-a-aspartyl]-3-phenyl-L-alanine. ;8 ;Other examples of compounds with formula I include the following: ;N-[N-[N-(l-amidino-4-piperidinylcarbonyl)-f3-alanyl]-L-a-aspartyl]-3-phenyl-L-alanine, ;N-[N-[N-(p-amidinophenylacetyl)-p-alanyl]- ;L-a-aspartyl]-3-phenyl-L-alanine, ;N-[N-[4-(p-amidinophenylcarbamoy1)-butyl1]-L-a-asparty1]-3-phenyl1-L-alanine, ;N-[N-[(p-amidinophenylcarbamoy1)acety1]-10 L-a-aspartyl]-3-phenyl-L-alanine, ;rac-N-[l-(p-amidinobenzoyl)-3-piperidinyl-carbonyl]-f3-alanine, ;N-[4-(p-amidinobenzamido)butyryl]-p- ;alanine, ;15 N-[(DL)-N-(p-amidinobenzoyl)-3-phenyl-3- ;alanyl]-3-alanine, ;N/N'-C[(S)-(p-amidinobenzamido)ethylen] dicarbonyl]di-p-alanine, ;N2-(p-amidinobenzoyly-N4-(2-carboxyethyl)-20 L-asparagine, ;N-[5-(p-amidinobenzamido)valeryl]-3- ;alanine, ;rac-N-[[1-C 3-(l-amidino-4-piperidiny1)pro-pionyl]-3-piperidiny1]carbony1]-3-alanine, 25 N-[[(S)-l-(p-amidinobenzoyl)-2-pyrrolidinyl]- ;acetyl]-3-alanine, ;(S)-3-[[N-(p-amidinobenzoyl)-3~alanyl]~ amino-3-[(p-methoxyphenethyl)carbamoyl]propionic acid, ;N-[[(R)-l-(p-amidinobenzoyl)-3-pyrrolidinylj-30 carbonyl]-3-alanine, ;I ;9 ;α-amide of N-[N-(p-amidinobenzoyl)-2-methyl-3-alanyl]-L-aspartic acid, ;N-[N-[(p-amidinophenyl)acetyl]-p-alanyl]- ;p-alanine, ;5 rac-N-[[1-(p-amidinophenyl)acetyl]-3-pipe- ;ridinyl]-carbonyl]-3-alaninebenzylester, ;rac-N-[[1-[(p-amidinophenyl1)acety1]-3-piperidinyl]-carbonyl]-3-alanine, ;N-[N-[N-[3-(l-amidino-4-piperidinyl)propio-10 nyl]-0-alanyl]-L-α-aspartyl]-3-phenyl-L-alanine, ;Acid (S)-3-[[DL-N-(p-amidinobenzoyl)-3-methyl-3-alany1]amino]-γ-oxo-l-pyrrolidinebutyric, • ;DL-N-[N-(p-amidinobenzoyl)-3-alanyl]-3-methyl1-3-alanine, 15 N-[DL-N-(p-amidinobenzoyl)-2-pheny1-3- ;alanyl]-3-alanine, ;DL-N-[N-(p-amidinobenzoyl)-g-alanyl]-2-methyl-3-alanine, ;DL-N-[N-(p-amidinobenzoyl)-3-alanyl]-2-20 phenyl-3-alaninef ;p-C 2-[[N-[N-(p-amidinobenzoyl)-3~ acid alany1]-L-a-aspartyl]amino]ethyl1]benzoic acid, ;DL-N-CN-(p-amidinobenzoyl)-3-alanyl-3-phenyl-;3-alanine, ;25 1^ p-[ 2-[[N-[N- (p-amidinobenzoyl)-3~ ;alany1]-L-a-aspartyl]amino]ethyl1]phenoxy]acetic acid, ;Acid 1-[N-(p-amidinobenzoyl)-3-alanyl]- 4-piperidinacetic acid, 4-[N-(p-amidinobenzoyl)-3-alanyl-30 1-piperidinacetic acid. 10 According to the invention, the above compounds can be prepared by cleaving at least one protecting group from a compound of formula x'-y-co-z'-ch(q1)-coo-q5 13 where Q1 and Y have the meanings given above, X' is a phenyl group substituted at position 4 by an amidino group, possibly protected, or a 4-piperidinyl, and Z' or Q has the same meaning as given above for Z or Q, or b) in a nitrile of formula c-O-y-co"2 "c H^" cûo-q' by transforming the nitrile group into the amidino group and if This is desired by transforming a compound of formula I into a physiologically acceptable salt, or a salt of a compound of formula I into the free acid or base. Examples of protecting groups in compounds of formula II include benzyl or lower alkyl groups, such as β-butyl, in ester groups such as benzyl-OCO- or β-Bu-OCO-, as well as amidino protecting groups, such as Z and Boc. Examples of protected amidino groups include -C(NH)NH-Z, -C(NH)NH-Boc, and -C(N-Boc)-NH-Boc. Ester groups can be cleaved in a known manner, for example, by saponification with a base, such as an alkali metal hydroxide, e.g., sodium hydroxide, in a solvent, such as methanol. Benzyl esters can be separated by hydrogenation in the presence of a noble metal catalyst, such as 15 palladium on carbon (Pd/C) in a solvent, such as methanol, ethanol, formic acid, or acetic acid, at a temperature up to about 40°C, preferably at room temperature. Simultaneously, an amidino protecting group 20, such as Z, contained in group X', is then separated. Ester groups, such as β-butyl, or amidino protecting groups, such as Boc, e.g., with an acid such as formic acid or trifluoracetic acid, if desired, 25 in a solvent such as dichloromethane, at a temperature up to 40°C, preferably at room temperature. The compounds of formula II are new and are also the subject of the invention. Their preparation 30 can be carried out starting from known compounds by known processes, such as those described, for example, below. 12 Thus, in a first step or couple according to known processes, a Amino acid of formula ;H-Z'-CH(Q1)COO-Q22 IV ;with an acid of formula ;R5-NHCH(Q3l)(CH2)1_3-COOH Va ;R5-NHCH2CH(Q4)-COOH Vb ;^ C00H ;r . -COOH ;8- - 'Ois-) ;O ;or ;Vc ;Vd ;H-W-\---CHvCOOH ve ;^-tCH;)n-NHCO(CH2)B-COOH VI ;13 ;where ; is an easily separable alkyl group, is a hydrogen, methyl, phenyl, lower -COO-alkyl or -CINH(CH2 )2-C0-alkyl, ; is an amino protecting group, such as Z or Boc, ;R ;5 ;n = 0 and p = 3 or n = 2 and p = 1, ;with formation of an amide group. ;One can thus perform the Coupling of IV with Va, Vb, or VI, e.g., in tetrahydrofuran (THF) between -10°C and room temperature, under argon in the presence of O-benzotriazolyl-N,N,N',N'-tetramethyluronium hexafluorophosphate (HBTU). Coupling of IV with Vc, Vd, or Ve is carried out, for example, by first activating the acid Vc, Vd, or Ve in THF with chlorodimethoxytriazine and N-methylmorpholine, and then reacting the p-toluenesulfonate of amine IV with N-methylmorpholine. From the reaction product thus obtained, the protecting 5-amino group R contained therein, e.g., Z or Boc as described above, can then be selectively separated by catalytic hydrogenation or, as appropriate, by means of trifluoroacetic acid. An amine of formula ;H2NCH(Q31)(CH2)1_3 ;-CO-Z'-CH(Q1)COO-Q22 VIII ;14 ;obtained in this way, e.g., starting from IV and Va, with 4-cyanobenzoic acid or 4-cyanophenylacetic acid, to give a nitrile, e.g., a nitrile of formula III, e.g., as above for the coupling of IV with Va. ;A nitrile thus obtained, or a nitrile obtained by coupling IV and VI, e.g., by reaction with sulfuric acid and triethylamine in pyridine to give the thioamide, methylation with methyl iodide in acetone, and reaction with ammonium acetate in methanol, can be transformed into a compound II in which X' contains a free amidino group. Similarly, a nitrile III can be transformed into the corresponding compound of formula I. ;On can also react an amine of formula VIII above, e.g., with l-amidino-4-piperidinecarboxylic acid, p-amidinophenacetyl chloride, or p-amidinobenzoyl chloride to give a compound II where X is an l-amidino-4-piperidineyl or a p-amidinophenyl. Furthermore, an amine of formula VIII, or an amine obtained from a compound of formula IV and an acid of one of the formulas Va to Ve, in methylene chloride and an aqueous solution of sodium bicarbonate, can be reacted with p-amidinobenzoyl chloride and then, in the presence of sodium carbonate, with benzyl chloroformate, or, as the case may be, di-t-butyl dicarbonate, to give a compound of formula II in which X' is a p-amidinophenyl protected by Z or, as the case may be, Boc. The compounds of formulas IV-VI are known or can be prepared by analogy with known compounds or as stated in the following examples. ;One can thus prepare a nitrile VI by coupling the corresponding amine of formula NCC^H.(CH„)-NH0 with the corresponding acid of formula H00C(CH9)-COOQ, e.g., in a manner analogous to the coupling of an amine IV with an acid V, followed by cleavage of the ester group Q. To prepare a compound of formula II where X' is a 4-piperidinyl substituted at position 4 by a protected amidino group, one can first react a compound of formula in t-butanol and sodium hydroxide, e.g., with N,N'-bis(t-butoxycarbonyl)-S-methylisothiourea to give a compound of formula BocC.OOH, and then couple this latter, e.g., with a compound. ;of formula h2nch(q31)(ch2)1_3-conhch2ch2cooch2c6h5 Xld or fçj ;HNK—CH, CONHCH, CH, COOC4c xie ;• C rl^COfJ UCUL CH^tOOCVc, ;where 31 ;Q is a hydrogen, methyl, phenyl, -COO- ;lower alkyl or -CONH(CH2)2-COO-lower alkyl. ;5 A compound of formula II can also be prepared where X' is a phenyl or a 4-piperidinyl substituted at position 4 by a protected amidino group, by coupling an acid of formula X'-Y-COOH with an amine of formula IV above. 10 The compounds of formula I, their solvation products and their salts inhibit both ;1 ;17 ;the binding of fibrinogen, fibronectin and von Willebrand factor to the fibrinogen receptor of blood platelets (glycoprotein IIb/IIIa), and the binding of these compounds and other proteins 5 Adhesives, such as vitronectin, collagen, and laminin, bind to corresponding receptors on the surface of various cell types. These compounds therefore influence cell-cell and cell-matrix interactions. They particularly inhibit the formation of blood platelet clots and can be used in the treatment or prevention of diseases such as thrombosis, stroke, myocardial infarction, inflammation, and arteriosclerosis. Furthermore, these compounds have an effect on tumor cells, inhibiting their metastasis. They can therefore also be used as antitumor agents. In addition, they can accelerate wound healing. The inhibition of fibrinogen formation at fibrinogen receptors by glycoprotein IIb/IIIa can be demonstrated as follows: Glycoprotein IIb/IIIa is obtained from extracts of Triton X-100 of human blood platelets-25 is purified by lectin affinity chromatography (Analytical Biochemistry 151, 1985, 169-177) and Arg-Gly-Asp-Ser affinity column chromatography (Science 231, 1986, 1559-62). The resulting receptor protein is fixed onto microtiter plates. The specific binding of fibrinogen to the immobilized receptors is determined using an ELISA (enzyme-linked immunosorbent assay). The Clgg values below correspond to the concentration of the experimental substance required to inhibit fibrinogen binding to the immobilized receptor by 50%: Example product: 1 2 3 4 5 6 7 Cl5Q (M) 0.0003 0.13 0.045 0.035 0.01 0.01 0.027; Product of the example: ; 8 9 10 11 18 25 28 31 38 39 ; 15 ; CI50 (mM) ; 0.10 0.005 0.022 0.0025 0.010 0.0067 0.005 0.0128 0.0036 0.0012 ; 20 As mentioned above, medicinal products containing a compound of formula I, one of their solvation products or one of their salts, are also the subject of the invention, as well as a method for preparing these medicinal products, which is characterized in that 25 one or more of the aforementioned compounds are placed in a galenic administration form, and if desired, one or several other therapeutically interesting substances. The drugs can be administered enterally, e.g., orally in the form of tablets, coated tablets, dragees, hard and soft gelatin capsules, solutions, emulsions, or suspensions, or rectally, e.g., as suppositories or as a sprayed liquid. However, administration can also be carried out parenterally, e.g., as injectable solutions or by infusion. To prepare tablets, coated tablets, dragees, and hard gelatin capsules, the active substance can be mixed with pharmaceutically inert inorganic or organic excipients. Examples of such excipients for tablets, dragees, and hard gelatin capsules include lactose, maize starch or a derivative thereof, talc, stearic acid, or its derivatives. For capsules of Soft gelatin is suitable as an excipient, e.g., vegetable oils, waxes, fats, and semi-solid and liquid polyols; however, depending on the nature of the active substance, it may not be necessary to use an excipient at all for soft gelatin capsules. For the preparation of solutions and syrups, excipients may include, e.g., water, polyols, sucrose, invert sugar, and glucose; for injectable solutions, e.g., water, alcohols, polyols, glycerin, and vegetable oils; and for suppositories, e.g., natural or hardened oils, waxes, fats, and semi-liquid or liquid polyols. Pharmaceutical preparations may also contain preservatives, solvents, stabilizing agents, wetting agents, emulsifying agents, softeners, and colorants. aromatics, salts to modify osmotic pressure, buffers, coating agents or antioxidants.

20

To combat or prevent the diseases mentioned above, the dose of the active substance can be varied within a wide range and must, of course, be adapted to the individual circumstances of each case. Generally, for oral administration, a dose of approximately 0.1 to 20 mg/kg should be used, preferably approximately 0.5 to 4 mg/kg per day for an adult.

but the upper limit just indicated can also be exceeded when it proves appropriate.

Example 1

In a suspension of 100 mg of N-[N-[N-(p-amidinobenzoyl)-3-alanyl]-3-(t-butoxycarbonyl)-L-alany1]-3-phenyl-L-alanine t-butyl ester iodide in 10 mL of dichloromethane, 15 mL of trifluoracetic acid is added. After 3 hours at room temperature, the solvent is evaporated and the residue is crystallized with ether. After recrystallization from methanol/ethyl acetate, 32 mg of N-[N-[N-(p-amidinobenzoyl)-3-alanyl]-L-α-aspartyl]-3-phenyl-L-alanine trifluoracetate is obtained, Pf. 216–220°C (decomposition).

The starting product can be prepared as follows:

a) In a solution of 446 mg of Z-3~Ala-

OH and 785 mg of H-Asp(O-tBu)-Phe-O-tBu (obtained from the condensation of Z-Asp(O-tBu)-OH with H-Phe-O-tBu followed by hydrogenolysis) in 15 mL of THF are added under argon at 0°C to 834 mg of HBTU and 0.24 mL of N-methylmorpholine. After 5 hours, the residue is concentrated and dispersed on acetate.

21

5% ethyl/NaHCO₃. Wash with water and 1M KHSO₄. Dry the organic extracts, filter and concentrate. Recrystallize the residue from ethyl acetate/diisopropyl ether, yielding 5592 mg of Z-3-Ala-Asp(0-tBu)-Phe-0-tBu, Pf. 138-139°C.

b) By catalytic hydrogenation of the intermediate product (550 mg) in ethanol in the presence of 10% Pd/C, the following is obtained after silica gel chromatography with ethyl acetate/methanol

10 4:1->1ï1 417 mg of H-3-Ala-Asp(O-tBu)-Phe-O-tBu, SM: 464 (M+H)+.

c) By coupling 350 mg of the product of b)

with 125 mg of 4-cyanobenzoic acid in the manner described in example a), after chromato-

15 Silica gel graph with ethyl acetate 293 mg of t-butyl ester of N-[3-(t-butoxycarbonyl)-N-[N-(p-cyanobeneoyl)-3-alanyl]-L-alanyl]-3-phenyl-L-alanine, Pf. 74-76°C.

d) A 20 solution of 270 mg of the intermediate product in pyridine/triethylamine 15:1 is saturated with sulfuric acid. After 24 hours, the solvent is removed and the residue is dispersed on 5% ethyl acetate/NaHCO3. The organic extracts are washed with water and with a 1 M 25 solution of potassium bicarbonate, dried, and concentrated. After silica gel chromatography of the residue with ethyl acetate, followed by recrystallization from hexane, 230 mg of β-butyl ester of N-[3-(β-butoxycarbonyl)-30 N-[N-[p-thiocarbamoylbenzoyl]-3-alanyl]-L-alanyl]-3-phenyl-L-alanine is obtained, Pf. 101-103°C.

22

e) In a solution of 200 mg of the intermediate product in 15 ml of acetone, 2 ml of methyl iodide is added. After 3 hours at boiling point, the mixture is allowed to cool to room temperature and the product is precipitated by the addition of diethyl ether. 226 mg of t-butyl ester iodide of N-[3-(t-butoxycarbonyl)-N-[N-[p-[1-(methylthio)formimidoylbenzoyl]-3-alanyl]-L-alanyl]-3-phenyl-L-alanine is obtained, at boiling point 139-140°C.

10 f) In a solution of 160 mg of the intermediate in 10 mL of methanol, 32 mg of ammonium acetate is added. The reaction mixture is maintained at boiling point for 4 hours. After cooling to room temperature, the solution is filtered, concentrated, and mixed with diethyl ether. The precipitate is separated by filtration and dried. 135 mg of β-butyl ester iodide of N-[N-[N-(p-amidinobenzoyl)-0-alanyl]-3-(β-butoxycarbonyl)-L-20 alanyl]-3-phenyl-L-alanine is obtained. Pf. 162-163°C.

Example 2

From 60 mg of t-butylester of n-[n-[n-(l-amidino-4-piperidinylcarbonyl)-0-alanyl]-3-(t-butoxycarbonyl)-L-alany1]-3-phenyl-L-alanine 25, treatment with trifluoracetic acid in methylene chloride, as stated in Example 1 after crystallization from methanol/diethyl ether, yields 26 mg of trifluoracetate of n-[n-[n-(l-amidino-4-piperidinyl-3 0 carbony1)-p-alany1]-L-a-aspartyl]-3-phenyl-L-alanine (2:3), Pf. 127-130°C.

23

The starting product is prepared as follows:

The coupling of 86 mg of 1-amidino-4-piperidinecarboxylic acid (Belgian Patent No. 893,282, 5 Nippon Chemiphar Co. Ltd.) with 119 mg of

H-3-Ala-Asp(O-tBu)-Phe-O-tBu (example lb) in dioxane in the presence of 58 mg of pyridinium hydrochloride, in the manner described in example la) gives after silica gel chromatography 10 with methylene chloride/methanol 1:1 71 mg of N-[N-[N- ( l-amidino-4-piperidinylcarbony 1 ) -(3-alany 1 ]-3-(t-butoxycarbonyl)-L-alany1]-3-pheny1-L-alany1-t-butylester, SM : 617 (M+H)+.

Example 3

15 From 50 mg of t-butyl ester iodihydrate of N-[N-[N-(p-amidinophenylacetyl)-3-alanyl]-3-(t-butoxycarbonyl)-L-alanyl]-3-phenyl-L-alanine, treatment with trifluoroacetic acid as stated in Example 1 yields 36 mg of 20 trifluoroacetate of N-[N-[N-(p-amidinophenylacetyl)-3-alanyl]-L-a-aspartyl]-3-phenyl-L-alanine, Pf. 218-220°C (from the diethyl ether).

The starting product can be prepared as follows:

25 a) By coupling 142 mg of 4-cyano- acid

phenylacetic acid with 371 mg of H-3-Ala-Asp(O-tBu)-Phe-O-tBu (example lb) is obtained in the manner described in example la) after silica gel chromatography with ethyl acetate/methanol 30 250 mg of t-butyl ester of N-[N-[N-(p-cyanophenyl-acetyl)-3-alanyl]-3-t-butoxycarbony1-L-alany1]-3-phenyl-L-alanine, Pf. 93-94°C.

24

b) Thionation of 230 mg of the intermediate product as stated in example ld) gives after silica gel chromatography with ethyl acetate, 160 mg of t-butyl ester of N-[N-[N-(p-thiocarbamoylphenylacetyl)-3-alanylJ-3-t-butoxy-carbonyl-L-alanyl]-3-phenyl-L-alanine, SM: 607 (M+H)+.

c) Methylation of 120 mg of the intermediate product in a manner analogous to example (e)

after crystallization gives 100 mg of t-butyl ester iodide of N-[N-[N-(p-methylthioformimidoyl)-phenyl)acetyl]-3-alanyl]-3-t-butoxycarbonyl-L-alany1]-3-phenyl-L-alaninef Pf. 103-104°C (acetone/diethyl ether)

d) The reaction of 90 mg of the intermediate product with ammonium acetate as stated in example lf) gives 62 mg of t-butyl ester iodide N-[N-C N-(p-amidinophenylacetyl)-3-alanyl]-3-t-butoxy-carbonyl-L-alanyl]-3-phenyl-L-alanine, Pf. 152°C (methanol/diethyl ether).

Example 4

By treating 55 mg of t-butylester iodide of N-[N-[4-(p-amidinophenylcarbamoyl1)-butyryl]-3-(t-butoxycarbonyl)-L-alanine1]-3-phenyl-L-alanine with trifluoracetic acid as stated in Example 1, 31 mg of N-[N-[4-(p-amidinophenyl1-carbamoyl)butyryl1]-L-a-aspartyl]-3-phenyl-L-alanine (5:4) trifluoracetate is obtained after crystallization from ethanol/diethyl ether, Pf. 159-161°C.

The starting product can be prepared as follows:

a) By coupling 1.18 g of 4-aminobenzo-

25

nitrile with 1.25 ml of monomethylglutarate as stated in example a)/ after recrystallisation from ethyl acetate 1.83 g of methyl 4-[(p-cyanophenyl)carbamoyl]butyrate is obtained, Pf. 126-127°C.

b) In a solution of 1 g of the intermediate in 10 mL of methanol, 6 mL of sodium hydroxide solution (IN) is added. After 7 hours, the reaction mixture is concentrated, extracted with ethyl acetate, and then neutralized with hydrochloric acid (IN). The resulting precipitate is separated by filtration, washed with water, and dried. 575 mg of 4-β-C(p-cyanophenyl)carbamoylbutyric acid (Pf. 200-201°C) is obtained.

c) By coupling 464 mg of the intermediate product with 863 mg of H-Asp(0-tBu)-Phe-0-tBu as stated in example a), we obtain after silica gel chromatography with ethyl acetate and then by diluting in hexane 701 mg of t-butyl ester of N-[3-(t-butoxycarbonyl)-N-C4-(p-cyano-phenylcarbamoyl)-butyryl]-L-alanyl]-3-phenyl-L-alanine Pf. 64-66°C.

d) The reaction of 350 mg of intermediate product with sulfuric acid by analogy with example ld) after recrystallization with ethyl acetate gives 304 mg of t-butyl ester of N-[3-(t-butoxycarbonyl)-N-[4-(p-thiocarbamoyl-1-phenyl)butyryl]-L-alanyl]-3-phenyl-L-alanine,

Pf. 178-179°C.

e) Methylation of 150 mg of the intermediate product, by analogy with example le), gives, after crystallization with diethyl ether, 175 mg

27

hydrochloric acid IN. The THF is evaporated and the aqueous extracts are separated with ethyl acetate. After drying and concentration, 380 mg of methyl-(p-cyanophenethyl)malonamate, Pf. 125°C, are obtained. 5 c) Analogously to example 4b), the saponification of 750 mg of the product of b) gives 365 mg of N-(p-cyanophenethyl)malonamic acid, Pf. 137-139°C.

d) In a manner analogous to example a), 10 is obtained by coupling 300 mg of the intermediate product with 507 mg of H-Asp(O-tBu)-Phe-O-tBu after silica gel chromatography with ethyl acetate, 352 mg of t-butyl ester of N-[3-(t-butoxycarbonyl1)-N-[(p-cyanophenethylcarbamoyl1)-acety1]-

15 L-alanyl]-3-phenyl-L-alanine, MS: 607 (M+H)+.

e) In a manner analogous to example ld), e)

and f), this same reaction sequence gives with 380 mg of the intermediate product of d) after crystallization with diisopropyl ether, 152 mg of 20-t-butyl ester iodihydrate of N-[N-[(p-amidinophenethylcarbamoyl D-acetyl-3-(t-butoxycarbonyl)-L-alanyl]-3-phenyl-L-alanine, Pf. 91-93°C (decomposition).

Example 6

From 200 mg of t-25 butylester iodihydrate of N-[N-[4-(p-amidinobenzamido)butyryl]-3-(t-butoxycarbonyl)-L-alanyl]-L-valine, we obtain by analogy with example 1 after recrystallization from ethanol/diethyl ether, 119 mg of trifluor acetate of N-[N-[4-(p-amidinobenzamido)butyryl]-L-30 α-aspartyl]-L-valine, Pf. 174°C (decomposition).

The starting product can be prepared as follows:

c

26

t-butylester iodihydrate of N-[3-(t-butoxycarbonyl)-N-C4-[Cp-1-(methylthio)-formimidoyl]phenyl]-carbamoy1]butyryl]-L-alany1]-3-phenyl-L-alanine (1:1), Pf. 127-128°C.

5 f) From 90 mg of the intermediate product,

by analogy with example lf) we obtain 60 mg of t-butyl ester iodide of N-CN-[4-(p-amidinophenyl lcarbamoyl)-butyryl]-3-(t-butoxycarbonyl)-L-alanyl]-3-phenyl-L-alanine, Pf 130-132°C.

10 Example 5

The reaction of 100 mg of N-CN-C(p-amidinophenethylcarbamoyl)-acetyl-3-(t-butoxycarbonyl)-L-alanine-3-phenyl-L-alanine t-butyl ester iodide with trifluoracetic acid as it

15 is said in example 1 gives after crystallization with diethyl ether 69 mg of trifluoracetate of N-CN-C(p-amidinophenethylcarbamoyl)-acetyl]-L-a-aspartyl]-3-phenyl-L-alanine, Pf. 141-143°C.

The starting product can be prepared as follows:

20 follows:

a) From 876 mg of p-cyanohydrocinnamic acid (Pharmazie 28, 1973, 724) one obtains, according to a known reaction sequence (Organic Synthesis 51, 1971, 48) 370 mg of p-(2-aminoethyl)-

25 benzonitrile, SM: 147 (M+H)+.

b) In a solution of 731 mg of p-(2-aminoethyl)benzonitrile and 1.39 ml of triethylamine in 10 ml of THF, a solution of 0.59 ml of methyl ester of the acid is added dropwise at -10°C

30 malonic acid in 5 ml of THF. Allow to warm to room temperature, pour into ice water and adjust to pH 2 with acid.

1

28

a) By coupling 783 mg of Z-4-amino-butyric acid with 1 g of H-Asp(O-tBu)-Val-O-tBu (obtained from the condensation of Z-Asp(O-tBu)-OH with H-Val-O-tBu followed by hydrogenolysis) as stated in example la), after silica gel chromatography with ethyl acetate and crystallization with hexane, 1.17 g of t-butyl ester of N-[N-[4-[l-(benzoyloxy)formamido]-butyramido]-3-(t-butoxycarbonyl)-L-alanyl]-L-valine is obtained, Pf. 64-65°C.

b) Hydrogenolysis of -1.1 g of the product of the intermediate step analogous to example b)

gives 1 g of t-butylester of N-[N-(4-aminobutyryl)~ 3-(t-butoxycarbonyl)-L-alanyl]-L-valine, Pf. 99-100°C.

c) In a solution of 324 mg of 4-cyanobenzoic acid, 352 mg of 2-chloro-4,6-dimethoxy-

1,3,5-triazine and 0.24 ml of N-methylmorpholine in 10 ml of dimethylformamide, after 2 hours at 0°C a solution of 859 mg of the product of b) is poured. It is allowed to warm to room temperature, then after purification as in example ld) and after silica gel chromatography with ethyl acetate, 772 mg of t-butyl ester of N-[N-C4-(p-cyanobenzamido)butyryl]-3-(t-butoxycarbonyl)-L-alanyl]-L-valine, SM: 559 (M+H)+, is obtained.

d) If 760 mg of the product of c) is administered

Following the same reaction sequence as in examples ld, e) and f), after crystallization with diisopropyl ether, 444 mg of t-butyl iodide-

29

ester of N-[N-[4-(p-amidinobenzamido)butyryl]-3-(t-butoxycarbonyl)-L-alanyl-L-valine, Pf. 105-107°C (decomposition).

Example 7

1.09 g of rac-N-[-[[[p-(benzyloxy-carbonyl)amidino]-benzoyl]-3-piperidinyl1]carbony1]-3-alaninebenzyl ester and 0.25 g of Pd/C are stirred in 20 mL of acetic acid under hydrogen. The catalyst is separated by filtration and the filtrate is concentrated. The residue is absorbed into water and concentrated. The precipitate is suspended in ethanol.

We adjust it to pH 8 with ammonia and shake it, '

then we filter through a filtering funnel, wash with methanol and dry. We obtain 530 mg of rac-N-[[1-(p-amidinobenzoyl)-3-piperidinyl]-carbonyl]-3-alanine in the form of the hydrate (2:1), Pf >265°C, SM 347 (96, M+H).

The starting product can be prepared as follows:

a) Rac-N-(t-butoxycarbonyl)-piperidin-3-carboxylic acid (Can. J. Physiol. Pharmacol. 57, 1979, 763) is activated in THF with chlorodimethoxytriazine and N-methylmorpholine, then combined with 3-alaninebenzylester-p-toluenesulfonate

(J. Org. Chem. 17, 1952, 1564) and of N-methylmorpholine to rac-N-[[1-(t-butoxycarbonyl)-3-piperidinyl1]-carbonyl]-f3-alaninebenzylester, Pf. 57-59°C.

b) By cleavage with trifluoracetic acid, benz-yl-ester of rac-(3-piperidinylcarbonyl)-3-alanine is obtained from this trifluoracetate.

c) The latter is reacted in methylene chloride/aqueous bicarbonate solution

30

sodium with p-amidinobenzoyl chloride and then in the presence of sodium carbonate with benzyl chloroformate to give rac-N-l-[[[p-[N-(benzyloxycarbony1)amidino]benzoy1]-3-piperidiny1]-5 carbonyl]-3-alaninebenzylester, SM: 571 (10, M+H).

Example 8

512 mg of N-[4-[p-[N-(benzyloxy-carbonyl)amidino]benzamido]butyryl]-3-alaninebenzyl ester and 170 mg of Pd/C are stirred in 10 mL of acetic acid under hydrogen. The catalyst is separated by filtration.

The residue is dissolved in water and concentrated. The residue is suspended in water, adjusted to pH 7 with ammonia, filtered through a filtering funnel, washed with water, and dried. The result is...

239 mg of N-[4-(p-amidinobenzamido)butyryl]-3-alanine, m.p. >250°C, SM 321 (12, M+H).

The starting product can be prepared as follows:

20 a) 4-(t-butoxycarbonyl- acid is activated

amino)butyric with chlorodimethoxytriazine and N-methylmorpholine in THF and reacted with 3-alaninebenzylester-p-toluenesulfonate in the presence of N-methylmorpholine to give N-[4-(1-t-butoxyformamido)butyryl]-3-alaninebenzylester, Pf. 54-55°C.

b) In trifluoracetic acid, the trifluoracetate of N-(4-aminobutyryl)-3-alaninebenzyl ester is obtained from this. 30 c) The latter is reacted in methylene chloride/water/sodium bicarbonate with p-amidinobenzoyl chloride and then in the presence

31

of sodium carbonate with benzyl chloroformate to give N-[4-[p-[N-(benzyloxycarbonyl)-amidino]benzamido]butyryl]-3-alaninebenzylester, Pf. 173-183°C.

Example 9

Similar to example 8, from N-[N-[p-[N-(benzyloxycarbonyl)amidino]-benzoyl]-3-alanyl]-3-alaninebenzylester, N-[N-(p-amidinobenzoyl)-3-alanyl]-3-alanine, Pf. >250°C, SM: 307 (6, M+H), is obtained in the form of the hydrate (2:1).

The starting product can be prepared as follows:

a) N-(t-butoxycarbonyl)-3-alanine and 3-alaninebenzylester are coupled in a manner analogous to the examples above to give N-[N-(t-butoxycarbonyl)-3-alanyl]-3-alaninebenzylester, Pf. 84-85°C.

b) In trifluoracetic acid, trifluoracetate of (3-alanyl)-3-alaninebenzylester is obtained from this.

c) The latter is reacted with p-amidinobenzoyl chloride, then with benzyl chloroformate to give N-[N-[p-[N-(benzyloxycarbonyl)-amidino]benzoyl]-3-alanyl]-3-alaninebenzylester, Pf. 165-166°C.

Example 10

By analogy with example 8, from N-[(DL)-N-[p-[N-(benzyloxycarbonyl)amidino]-benzezoyl]-3-phenyl-3~alanyl]-3-alaninebenzylester, we obtain N-[(DL)-N-(p-amidinobenzoyl)-3-phenyl-3~alanyl]-3-alanine in the form of a hydrate (3:1), Pf. >250°C, SM: 383 (62, M+H).

32

The starting product can be prepared as follows:

a) We couple DL-N-(t-butoxycarbonyl)-3-phenyl-3-alanine and 3-alanin-benzylester in a manner analogous to the examples above to give N-[(DL)-N-(t-butoxycarbonyl)-3-phenyl1-3-alanin1]-3-alaninebenzylester, Pf. 143-144°C.

b) In trifluoracetic acid, we obtain

from this the trifluoracetate of [(DL)-3-phenyl-3-alanyl]-3-alaninebenzylester.

c) This is reacted with p-amidinobenzoyl chloride and then with benzyl chloroformate to give N-[(DL)-N-[p-[N-(benzyloxycarbonyl )amidino]benzoyl]-3-pheny1-3-alany1]-3-alaninebenzylester, Pf. 187-189°C.

Example 11

443 mg of N-[3-[(benzyloxy)carbonyl]-N-[5-[p-[N-[(benzyloxy)carbonyl]amidino]benzamido]-valeryl]-L-alanyl]-3-phenyl-L-alaninebenzylester and 111 mg of Pd/C are stirred in 9 mL of acetic acid for 3 hours % under a hydrogen atmosphere. The solution is filtered, concentrated, the residue is dissolved in water, and the solution is concentrated again. The residue is diluted in water, filtered through a filter funnel, and dried. 246 mg of N-[N-[5-(p-amidinobenzamido)valeryl]-L-α-aspartyl]-3-phenyl-L-alanine, Pf. 241°C, is obtained as the (1;2) hydrate.

The starting ester, Pf. 169-171°C, can be prepared as follows:

a) The 4-benzylester of the N- acid is coupled

(t-butoxycarbonyl)-L-aspartic acid with 3-phenyl-L-alanine benzyl ester to give N-[3-[(benzyloxy)-

33

carbonyl]-N-(t-butoxycarbonyl)-L-alany1]-3-phenyl-L-alanine benzylester, mp 93-94°C.

b) This is deprotected with tri-fluoracetic acid and coupled with 5-(1-t-butoxyformamido)valeric acid to give N-[3-

[(benzyloxy)carbonyl]-N-C 5-(1-t-butoxyformamido)-valeryl]-L-alany li]-3-phenyl-L-alaninebenzy lester, m.p. 119.5-120.5°C.

c) The latter is stripped of the protecting t-butoxycarbonyl group in tri-fluoracetic acid, then transformed into the starting product in methylene chloride/water/sodium bicarbonate with p-amidinobenzoyl chloride, then with benzyl chloroformate.

Example 12

Similar to Example 11, N,N'-[[(S)-[p-[N-[(benzoyloxy)carbonyl]-amidino]benzamido]ethylen]dicarbonyl]-di-3-alanine-dibenzyldiester is obtained after evaporation with water and dilution with methanol, yielding N,N'-[[(S)-(p-amidino benzamido)-ethylen]dicarbonyl]di-3-alanine, Pf. >250°

The starting ester, Pf. 171-172°C, is obtained as follows:

a) N-(t-butoxycarbonyl)-L-aspartic acid is coupled with two equivalents of 3-alaninebenzyl ester to give N,N'-[[(S)—(1-t-butoxyformamido) ethylen]dicarbonyl]di-p-alaninedibenzylester,

Pf. 109-110°C.

b) After cleavage of the t-butoxycarbonyl group with trifluoracetic acid, it is reacted in methylene chloride/water/sodium bicarbonate with p-amidinobenzoyl chloride and then

34

with benzyl chloroformate to give the starting ester.

Example 13 In a manner analogous to example 11, from N-[(S)-N-[p-[N-[(benzyloxy)carbonyl]-amidino]benzoyl]-3-[(benzyloxy)carbonyl]-3-alanyl]-3-alaninebenzylester, N2-(p-amidinobenzoyl)-N4-(2-carboxyethyl)-L-asparagine is obtained, Pf. 212°C (dec.).

The starting ester is obtained as follows:

a) We couple a benzylester of N-(t-butoxycarbonyl)-L-aspartic acid with 3-alaninebenzylester to give the 3-[[2—[(benzyloxy)carbonyl]-ethyl]carbamoyl]-N-(t-butoxycarbonyl)-L-alaninebenzyl-ester, Pf. 77-78°C.

b) After cleavage of the t-butoxycarbonyl protecting group in trifluoracetic acid, it is coupled in methylene chloride/water/sodium bicarbonate with p-amidinobenzoyl chloride and then reacted with benzyl chloroformate to give the starting ester, Pf. 122-123°C.

Example 14

In a manner analogous to example 8, from N-[5-[p-[N-[(benzyloxy)carbonyl]amidino]-benzamido]valeryl]-3~alanin-benzylester, N-[5-(p-amidinobenzamido)valeryl1]-3-alanine, Pf. >280°C.

The starting ester can be prepared as follows:

a) We couple 5-(1-t-butoxyformamido)-valeric acid with p-alanin-benzylester to give N-[5-(1-t-butoxyformamido)valeryl]-3~alanin-benzylester, Pf. 69-70°C.

b) In trifluoracetic acid, this yields N-(5-amino- trifluoracetate

35

valeryl)-3-alaninbenzylester.

c) The latter is reacted in methylene chloride/water in the presence of sodium bicarbonate with p-amidinobenzoyl chloride and then with benzyl 5-chloroformate to give the starting ester, Pf. 161-161.5°C.

Example 15

704 mg of rac-N-[[l-[3-[1-C(E/Z))-N,N'-bis(t-butoxycarbonyl)amidino]-4-piperidinyl1]-10 propionyl]-3-piperidinyl]carbonyl]-3-alaninebenzyl ester and 176 mg of Pd/C are stirred in 14.1 mL of formic acid for 18 hours under nitrogen. The catalyst is separated by filtration and washed with 1:1 formic acid-water. The filtrate is concentrated, residue 15 is dissolved in water, and the solution is concentrated again.

The residue is chromatographed on silica gel with ethanol-methanol. 254 mg of rac-N-[C1—C3—(l-amidino-4-piperidinyl)propionyl]-3-piperidinyl]carbonyl]-3-alanine is obtained, SM: 382 (100, M+H). The starting ester can be prepared as follows:

a) 4-piperidine-propionic acid is reacted in t-butanol and 2N sodium hydroxide with N,N'-bis(t-butoxycarbonyl)-S-methyl-

25 isothiourea to give the acid 3—[1— C(E/Z)-N,N'-

bis(t-butoxycarbonyl)amidino]-4-piperidinyl]propionic acid.

b) This compound is coupled with rac-N-(3-piperidinyl-carbonyl)-3-alaninebenzylester to give the starting ester, SM: 672 (12, M+H). Example 16

115 mg of N-[C(S)-l-[p-[N-[(benzyloxy)carbonyl]-amidino]benzoyl]-2-pyrrolidinyl]acetyl]-3-alanine and 50 mg of Pd/C are shaken in 2.5 ml of acetic acid for 4 hours under nitrogen

"h

36

for 4 hours. The catalyst is separated by filtration and the solution is concentrated. The residue is dissolved in water, concentrated again, and chromatographed with methanol on silica gel. 46 mg of N-[[(S)-l-(p-amidinobenzoyl)-2-pyrro-lidinyl]acetyl]-3-alanine, Pf. >250°C, is obtained.

The starting product is prepared as follows:

a) We couple the acid (S)-l-[(benzyloxy)carbonyl]-

2-pyrrolidinacetic acid with 3-alanin-t-butyl ester to give N-[[(S)-l-[(benzyloxy)carbonyl]-2-pyrrolidinyl]acetyl]-3-alanin-t-butyl ester,

SM: 391 (69, M+H).

b) By hydrogenation in acetic acid, the acetate of N-[(2-pyrrolidinyl)acetyl]-3~alanin-t-butyl ester is obtained from this.

c) This substance is reacted in methylene chloride/water in the presence of sodium bicarbonate with p-amidinobenzoyl chloride and then with benzyl chloroformate to give N-[[(S)- 1-C p-[N-[(benzyloxy)carbonyl]amidino]benzoy1]-2-pyrro-lidinyl]acetyl]-3-alanin-t-butylester, Pf. 127-128°C.

d) In formic acid, we obtain from this the starting compound, SM: 481 (100, M+H).

Example 17

300 mg of benzyl-(S)- is shaken under nitrogen

3-[[N-(t-butoxycarbonyl)amidino]benzoy1]-3-alanyl]-amino]-3-C(p-methoxyphenethyl1)-carbamoy1]propionate and 75 mg of Pd/C for 4 hours in 6 ml of formic acid. The catalyst is separated by filtration, the filtrate is concentrated, the residue is absorbed into water, and the solution is concentrated again. The crystalline substance is suspended in water.

37

It is adjusted to pH 8 with ammonia and then filtered with a filtering funnel. 151 mg of (S)-3-[[N-(p-amidinobenzoyl)-3-alanyl]amino-3-[(p-methoxyphenethyl)carbamoyl]propionic acid is obtained in the form of a hydrate (1:1), Pf. 217°C.

The starting ester can be obtained as follows:

a) The 3-alaninebenzyl ester is reacted in methylene chloride/water/bicarbonate of

10 sodium with p-amidinobenzoyl chloride then with di-t-butyl dicarbonate and sodium carbonate to give N-[p-[N-(t-butoxycarbonyl)-amidino]benzoyl]-3-alanin-benzylester, Pf. 127-128°C.

b) By catalytic hydrogenation, we obtain

15 from this the N-[p-[N-(t-butoxycarbonyl)amidino]-

benzoyl]-3-alanine, SM: 336 (21, M+H).

c) The 4-benzyl ester of N-(t-butoxycarbonyl)aspartic acid is coupled with 2-(4-methoxyphenyl)ethylamine to give t-butyl-[(S)-2-[(benzyl-

20 oxy)carbonyl3—1—[(p-methoxyphenethyl)carbamoyl]-ethyl]-

carbamate, Pf. 103-104°C.

d) In trifluoracetic acid, we obtain

from this the trifluoracetate of 3-[(p-methoxyphenethyl)carbamoyl]-3_alaninebenzylester. 25 e) The latter is coupled with the product described in b) to give the starting ester, Pf. 150°C, (dec.).

Example 18

Similar to example 8, we obtain from N-[3-[(benzyloxycarbonyl]-N-[N-[p-[N-30 C(benzyloxy)carbonyl]amidino]benzoyl]-3-alanyl]-L-

alanyl]-L-leucinisopropylester the N-[N-[N-(p-amidino-benzoy1)-3-alanyl]-L-a-aspartyl]-L-leucinisopropylester in the form of a hydrate (1:1:3),

Pf. 234°C (dec.).

38

The starting ester can be obtained in the following way:

a) N-(t-butoxycarbonyl)-L-leucine is transformed into pyridine, with dicyclohexyl-

5 carbodiimide/ of p-toluenesulfonic acid and isopropanol in the N-(t-butoxycarbonyl)-L-leuciniso-propylester, Ca]D = -33° (MeOH, c = 0.5).

b) In trifluoracetic acid, L-leuciniiso- trifluoracetate is obtained from this.

10 propylester.

c) This is coupled with the 4-benzyl ester of N-(t-butoxycarbonyl)-L-aspartic acid to give the N-[3-[(benzyloxy)carbonyl]-N-(t-butoxycarbonyl)-L-alanyl]-L-leucinisopropyl ester. SM: 479 (25, M+H). 15 d) After cleavage of the protecting t-butoxy- group

carbonyl and coupling with N-(t-butoxycarbonyl)-3-alanine, we obtain from this the N-[3-[(benzyloxy)carbonyl]-N-[N-(t-butoxycarbonyl)-3-alanyl]-L-alanyl]-L-leucinisopropylester, Pf. 103-104°C. 20 e) We remove the t-butoxycarbonyl protecting group in trifluoracetic acid and then transform it in methylene chloride/water/sodium bicarbonate with p-amidinobenzoyl chloride and then with benzyl chloroformate to 25 the starting ester, Pf. 176-177°C.

Example 19

Similar to example 8, N-[[(R)-l-[p-[N-[(benzyloxy)carbonyl]-amidino]benzoyl]-3-pyrrolidinyl]carbonyl]-3-alanine-30 benzylester is obtained as a hydrate (4:3), Pf. >250°C, [a]D = -4.13° (IN HC1, c = 0.46%)

39

The starting ester can be prepared as follows:

a) (R)-l-[(R)-a-methylbenzyl]-3-pyrrolidinemethanol, di-t-butyl dicarbonate and Pd/C are stirred for 20 hours in ethanol under hydrogen. The t-butyl ester of (R)-3-(hydroxymethyl)-l-pyrrolidinecarboxylic acid is obtained, Pf. 35°C [<x]D = +19.5° (methanol, c= 0.10).

b) From this, we obtain, with pyridinium dichromate in DMF, the acid

(R)-1-(t-butoxycarbonyl)-3-pyrrolidine,

Mp. 135-138°C, Ca]D = -15° (MeOH, c = 1.0).

c) This compound is coupled with (3-alaninebenzylester to give N-[C(R)-l-(t-butoxycarbonyl)-3-pyrro-lidinyl]carbonyl]-Σ-alaninebenzylester, Pf. 83-84°C, [<x]D = -3.4° (MeOH, c- 1.0)

d) In trifluoracetic acid, from this we obtain N-[[(R)-3-pyrrolidinyl]carbonyl]-O-alaninebenzyl ester trifluoracetate.

e) This substance is reacted in methylene chloride/water/sodium bicarbonate with p-amidinobenzoyl chloride and then with benzyl chloroformate to give the starting ester [a]D = +2.2° (MeOH, c= 0.5).

Example 20 Analogously to Example 16, we obtain from benzyl-(S)-3-[[D/L-N-[p-[N-[(benzyl-

I

oxy)carbonyl]amidino]benzoy1]-2-methyl-3-alany1]amino]-succinamate the α-amide of N-[N-(p-amidinoben-zoyl)-2-methyl-3-alanyl]-L-aspartic acid (Epimer 2:1). Pf. 280°C.

40

The starting ester can be prepared as follows:

a) Benzyl-(S)-3-(1-t-butoxy-formamido)succinamate is deprotected with trifluoracetic acid and then coupled with DL-N-(t-butoxycarbonyl)-

2-methyl-f3-alanine. We obtain t-butyl-[ (RS )-2-[[(S)-2-[(benzyloxy)carbonyl]-l-carbamoylethyl]-carbamoyl]propyl]carbamatef Pf. 135-136°C.

b) After cleavage of the t-butoxycarbonyl protecting group in trifluoracetic acid, it is coupled in methylene chloride/water/sodium bicarbonate with p-amidinobenzoyl chloride and then reacted with benzyl chloroformate to give the starting ester (2:1 epimer),

Pf. 178.5-179.5°C.

Example 21

548 mg of N-[N-[p-[N-(t-butoxycarbonyl)amidino]phenyl]acetyl]-

3-Alanyl-3-alanine benzyl ester in 11 mL of formic acid. After adding 137 mg of Pd/C, the mixture is stirred under hydrogen for 4 hours. The catalyst is separated by filtration and the filtrate is concentrated.

The residue is dissolved in water and the solution is concentrated again. The residue is suspended in water and adjusted to pH 8 with ammonia.

then it is filtered through a filtering funnel and dried. 290 mg of N-[N-[(p-amidinophenyl)-acetyl]-3-alanyl]-3-alanine is obtained as a hydrate (1:1), Pf. 286°C (dec.).

The starting product, Pf. 262°C (dec.), is obtained by reacting N-(3-alanyl)-3-alaninebenzylester in DMF/triethylamine with chloride

41

rte p-amidinophenylacetyl then by reaction with di-t-butyl dicarbonate.

Example 22 535 mg of rac-N-[[l-5 [p-[N-(t-butoxycarbonyl)amidino]-phenylacetyl]-3-

piperidinyl]carbonyl]-3-alaninebenzylester for 19 hours in 11 ml of formic acid. The solvent is evaporated, the residue is evaporated with water, and the mixture is recrystallized from acetonitrile. 340 mg of rac-N-[[[1-(p-amidinopheneDacetyl]-3-piperidinyl]carbonyl]-3-alaninebenzylester-formate (1:1) is obtained, boiling at 97-98°C.

The starting ester, SM: 551 (9, M+H), is obtained by coupling rac-N-[(3-piperidinyl1)carbonyl]-15 3-alaninebenzylester with p-amidino-phenylacetyl chloride in DMF/triethylamine and then by reaction with di-t-butyl dicarbonate.

Example 23. 20. 535 mg of rac-N-[[l-[p-[N-(t-butoxycarbonyl)amidino]-phenylacetyl]-3-piperidinyl]carbonyl]-3-alaninebenzyl1-ester is left to stand for 19 hours in 11 mL of formic acid, mixed with 134 mg of Pd/C, and stirred for 4 hours under nitrogen. The solution is filtered and concentrated; the residue is dissolved in water and the solution is concentrated again. The product is diluted in acetonitrile, filtered through a filtering funnel, and dried. 272 mg of rac-N-[Cl-C(p-amidinophenyl)-acetyl]-3-piperidinyl]carbonyl]-3-alanine are obtained, SM: 361 30 (41, M+H).

t

42

Example 24

1127 mg of N-[3-[(benzyloxy)carbonyl]-N-[N-[3-[l-[(E or Z)-N,N'-bis(t-butoxycarbonyl)amidino]-4-piperidinyl1]-propionyl]-3-alanyl]-L-alanyl]-3-phenyl-L-alaninebenzyl ester is left to stand in 22.5 mL of formic acid for 21 hours. After the addition of 282 mg of Pd/C, the solution is stirred for 5 hours under hydrogen. The solution is filtered and concentrated; the residue is dissolved in water and the solution is concentrated again. The residue is diluted in water, filtered through a filtering funnel, and dried. We obtain 543 mg of N-[N-[N-[3-(l-amidino-4-piperi- ' dinyl)propionyl]-3-alanyl]-L-a-aspartyl]-3-phenyl-L-alanine, Pf. 246°C (dec.).

The starting ester can be prepared as follows:

a) N-(t-butoxycarbonyl)-3-alanine is coupled with N-[3-[(benzyloxy)carbonyl]-L-alanyl]-3-pheny1-L-alaninebenzylester to give N-C3-[(benzyloxy)-carbonyl]-N-[N-(t-butoxycarbonyl)-3-alanyl]-L-alany1]-3-pheny1-L-alaninebenzylester, Pf. 124-125°C.

b) After cleavage of the β-butoxycarbonyl protecting group, it is coupled with the acid 3-[1-[N,N'-bis-

(t-butoxycarbonyl)amidino]-4-piperidinyl]propionic to give the starting ester, product of solvation of ethyl acetate 1:1, Pf 100°C (dec.).

Example 25

1.3 g of 3-[(benzyloxylcarbonyl]-N-[5-[p-[N-C(benzyloxy)-carbonyl]amidino]benzamido]-valeryl]-L-alaninisobutyl ester and 325 mg of Pd/C are stirred in 25 mL of acetic acid for 5 h under hydrogen. The solution is filtered and concentrated

43

The residue is evaporated successively with water, methanol, and ethanol. The product is adjusted to pH 8 in acetonitrile with ammonia, diluted, and filtered through a filtering funnel. 596 mg of isobutyl ester of N-[5-(p-amidinobenzamido)-valeryl]-L-α-aspartic acid is obtained as a hydrate (1:1) at temperatures of 162-166°C.

The starting ester, Pf. 127.5-129.5°C, can be prepared as follows:

a) The 4-benzylester of N-(t-butoxycarbonyl)-L-aspartic acid in pyridine is transformed with dicyclohexylcarboddimide, p-toluenesulfonic acid and isobutyl alcohol into the N-(t-butoxycarbonyl)-3-C(benzyloxy)carbonyl]-L-alaninisobutylester, SM: 323 (8, M-C^Hg).

b) After cleavage of the t-butoxycarbonyl group in trifluoracetic acid, it is coupled with 5-(1-t-butoxyformamido)valeric acid to give the 3-[(benzyloxy)carbonyl]-N-[5-(1-t-butoxy-formamido)valeryl]-L-alaninisobutylester, Pf.

64-66°C.

c) This is deprotected with tri- acid

fluoracetic then it is reacted in methylene chloride/water/sodium bicarbonate with p-amidinobenzoyl chloride and then with benzyl chloroformate to give the starting ester.

Example 26

562 mg of benzyl ester of (S)-3-[[DL-N-C p-C N-[(benzyloxy)carbonyl]amidino]-benzoyl]-3-methyl-3-alanyl]amino]-γ-oxo-l-pyrrolidin-butyric acid and 140 mg of Pd/C are stirred in 11 ml of acetic acid

44

for 2 hours under hydrogen. The filtered solution is concentrated and the residue is evaporated successively with water, methanol, and ethanol. Then, the pH is adjusted to 8 with ammonia in ethanol, the solution is diluted, and the mixture is filtered through a filtering funnel. 289 mg of (S)-f3-[[DL-N-(p-amidinobenzoyl)-3-methyl1-3-alany1]amino]-γ-oxo-1-pyrrolidinebutyric acid is obtained as a hydrate (2:3), Pf. 222-224°C.

The starting ester can be prepared as follows:

a) The benzylester is deprotected from the acid

(S)-3-(1-t-butoxyformamido)-y-oxo-l-pyrrolidinebutyric acid with trifluoracetic acid and it is coupled with (RS)-3-(l-t-butoxyformamido)butyric acid to give (S)-3-C(RS)-3-(1-t-butoxyformamido)-butyramido]-y-oxo-l-pyrrolidinebutyric acid, Pf. 104-105°C.

b) From this, after cleavage of the t-butoxycarbonyl group in trifluoracetic acid and reaction with p-amidinobenzoyl chloride and then with benzyl chloroformate in methylene chloride/water/sodium bicarbonate, we obtain the starting ester, SM: 642 (100, M+H).

Example 27

Similar to example 8, DL-N-[N-C(benzyloxy)carbonyl]amidino]-benzoyl]-3-alanyl]-3-methyl-3-alaninebenzylester is obtained in the form of a hydrate (3:1),

Pf. 291°C (dec.).

45

The starting ester, Pf. 179-180°C, can be prepared as follows:

a) N-(t-butoxycarbonyl)-3-alanine is coupled with the benzylester of DL-3-aminobutyric acid to give DL-N-(t-butoxycarbonyl)-(3-alanyl]-3-methyl-(3-alaninebenzylester, Pf. 70-72°C.

b) From this, DL-N-(3-alanyl)-3-methyl-3-alaninebenzylester is obtained from trifluoracetic acid.

c) We do; react this substance in methylene chloride/water/sodium bicarbonate with p-aminobenzoyl chloride and then with benzyl chloroformate to give the starting ester.

Example 28

In a manner analogous to example 8, from N-[3-[(benzyloxy)carbonyl°-N-[N-[p-[N-[(benzyloxy)carbonyl]amidinobenzoyl]-3-alanyl]-L-serinethyl ester is obtained the N-[N-[N-(p-amidinobenzoyl)-3~alanyl]-L-a-aspartyl]-L-serinethyl ester in the form of a hydrate (2:7) Pf. 201-203°C.

The starting product, Pf. 177-179°C, can be prepared as follows:

a) The 4-benzylester of N-(t-butoxycarbonyl)-L-aspartic acid is coupled with the L-serin-ethyl ester to give the N-[3-[(benzyloxy)carbonyl]-N-(t-butoxycarbonyl)-L-alany1]-L-serinethyl ester,

Pf. 96-97°C.

b) After cleavage of the t-butoxycarbonyl group, it is coupled with N-(t-butoxycarbonyl)-3-alanine to give N-[3-[(benzyloxy)carbonyl]-N-[N-(t-butoxycarbonyl)-3-alanyl]-L-alanyl]-L-serinethyl ester, Pf. 132-134°C.

46

c) This compound is deprotected in trifluoracetic acid and then reacted in methylene chloride/water/sodium bicarbonate with p-amidinobenzoyl chloride and finally with benzyl chloroformate to give the starting ester.

Example 29

In a manner analogous to example 8, from N-[N-[p-[DL-N-[(benzyloxy)carbonyl]-amidino]benzoyl]-2-phenyl-3-alanyl]-3-alaninebenzylester, N-[DL-N-(p-amidinobenzoyl)-2-phenyl-3~alanyl]-3-alanine is obtained in the form of a hydrate (4:1), Pf.

276°C.

The starting ester, Pf. 173-174°C, is prepared as follows:

a) DL-2-phenyl-3-alanine is reacted in t-butanol and sodium hydroxide with di-t-butyl dicarbonate to give DL-N-(t-butoxycarbonyl)-2-phenyl-3-alanine, Pf. 147-148°C.

b) This compound is coupled with 3-alaninebenzylester to give N-[DL-N-(t-butoxycarbonyl)-2-phenyl-3~alanyl]-3-alaninebenzylester, Pf. 115-116°C.

c) After cleavage of the protecting group, it is reacted in methylene chloride/water/sodium bicarbonate with p-amidinobenzoyl chloride and then with benzyl chloroformate to give the starting ester.

Example 30

Similar to example 8,

from N-[N-[p-[N-[(benzyloxy)carbonyl]-amidino]benzoyl]-3-alanyl]-DL-2-methyl-3-alaninebenzyl1-ester we obtain the DL-N-[N-(p-amidinobenzoyl)-3-alanyl]-2-

47

methyl-(3-alanine in hydrate form (3:1)

Pf. >300°C.

The starting ester, Pf. 159-160°C, is obtained as follows:

5 a) N-(t-butoxycarbonyl)-f3-alanine is coupled with [DL-2-methyl-p-alaninebenzylester] to give DL-N-[N-(t-butoxycarbonyl)-(3-alanine)-2-methyl1-3-alaninebenzylester, SM: 365 (47, M+H). b) After cleavage of the protecting group, we make

10. React in methylene chloride/water/sodium bicarbonate with p-amidinobenzoyl chloride and then with benzyl chloroformate to give the starting ester.

Example 31

15 In a manner analogous to example 11, from N-CN-C[(R)-l-[p-[N-[(benzyloxy)-carbonyl]amidino]benzoyl]-3-pyrrolidiny1]-carbonyl]-3-[(benzyloxy)carbonyl]-L-alany1]-3-pheny1-L-alanine-benzylester, after evaporation with water and dilution 20 in ethanol, N-[N-CC(R)-l-(p-amidinobenzoyl)-3-pyrrolidinyl]carbonyl]-L-a-aspartyl]-3-pheny1-L-alanin-acetate (2:1) is obtained in the form of a hydrate (3:2), Pf. 215°C.

The starting ester is obtained as follows: 25 a) The acid (R)-l-(t-butoxycarbonyl)- is coupled

3-pyrrolidinecarboxylic with N-C3-C(benzyloxy)-carbonyl]-L-alanyl]-3-phenyl-L-alanin-benzylester to give N-[N-[[(R)-l-(t-butoxycarbonyl)-3-pyrrolidiny1]carbonyl] —3— C(benzyloxy)carbonyl]-L-30 alanyl]-3-phenyl-L-alaninebenzylester, Pf. 84-85°C.

b) After removing the t-butoxy- group

S

48

carbonyl in trifluoracetic acid, is reacted in methylene chloride/water/sodium bicarbonate with p-amidinobenzoyl chloride and then with benzyl chloroformate to give the starting ester, Pf. 144-145°C.

Example 32 In a manner analogous to example 7, from the N-[N-[p-[N-[(benzyloxy)carbonyl]-amidino]benzoyl]-0-alanyl]-DL-2-phenyl-3-alaninebenzyl ester, the DL-N-CN-(p-amidinobenzoyl)-3-alanyl]-2-phenyl 3-alanine is obtained as a hydrate (1:1), Pf. 243-245°C.

The starting ester can be obtained as follows:

a) N-(t-butoxycarbonyl)-2-phenyl-3-alanine is boiled under reflux for 17 hours in acetone with benzyl bromide and potassium carbonate. DL-N-(t-butoxycarbonyl)-2-phenyl-3-alaninebenzylester is obtained, Pf. 58-59°C.

b) After cleavage of the t-butoxycarbonyl group, it is coupled with N-(t-butoxycarbonyl)-3-alanine to give DL-N-[N-(t-butoxycarbonyl)-3-alanyl]-2-phenyl-3~alaninebenzylester, Pf. 84.5-86°C.

c) This is transformed in tri-fluoracetic acid into trifluoracetate of -(3-alanyl)-2-phenyl-3-alaninebenzylester.

d) The latter is reacted in methylene chloride/water/sodium bicarbonate with p-amidinobenzoyl chloride and then with benzyl chloroformate to give the starting ester, Pf. 165-166°C.

49

Example 33 Analogously to Example 11, p-[2-C[3-[(benzyloxy)carbonyl]-N-[N-[p-[N-[(benzyloxy)carbonyl]-amidino]-benzoyl]-3-alanyl]-L-alany1]amino]ethy1]-benzoic acid is obtained from the benzylester of p-[2-[[N-[N-(p-amidinobenzoyl)-3-alanyl]-L-a-aspartyl]amino]ethy1]benzoic acid in the form of a hydrate (3:7), Pf. 194-196°C (methanol/water).

i

The starting ester, Pf. 162-163°C, can be prepared as follows:

a) p-(2-chloroethyl)benzoyl chloride is transformed in methylene chloride with benzyl alcohol and pyridine into the benzylester of p-(2-chloroethyl)benzoic acid, SM: 274 (8, M).

b) With sodium azide in DMSO

From this we obtain the benzylester of p-(2-azidoethyl)benzoic acid, SM: 281 (2, M).

c) This compound is reacted in pyridine with triphenylphosphine and then with concentrated ammonia to give the benzylester of p-(2-aminoethyl)-benzoic acid, SM: 226 (16, M-Cl^NH).

d) By coupling with the 4-benzylester of N-(t-butoxycarbonyl)-L-aspartic acid, we obtain from this the benzylester of p-[2-[[3-C(benzyloxy)carbonyl]-N-(t-butoxycarbonyl)-L-alany1]amino]-ethyl]benzoic acid, Pf. 99-100°C.

e) This is deprotected in tri-fluoracetic acid and coupled with N-(t-butoxy-carbonyl)-3-alanine to give the benzylester of p-[2-[[3- C(benzyloxy)-carbonyl]-N-[N-(t-butoxycarbonyl )-3-alanyl]-L-alany1]amino]ethy1]benzoic acid, Pf. 138-139 °C.

50

f) After cleavage of the t-butoxycarbonyl group in trifluoracetic acid, it is reacted in methylene chloride/water/sodium bicarbonate with p-amidinobenzoyl chloride and then with benzyl chloroformate to give the starting ester.

Example 34

In a manner analogous to example 7, from DL-N-[N-[p-[N-[(benzyloxy)carbonyl]-amidino]benzoyl]-(3-alanyl]-3-phenyl-1-3-alaninebenzyl ester, we obtain [DL-N-[N-(p-amidinobenzoyl)-3~alanyl-3-phenyl-p-alanine in the form of a hydrate (3:1), Pf. 220°C (dec.).

The starting ester is obtained, Pf. 208°C,

as follows:

a) N-(t-butoxycarbonyl)-3-alanine is coupled with DL-3-phenyl-3-alaninebenzylester to give DL-N-CN-(t-butoxycarbonyl)-3~alanyl]-3-phenyl-3-alaninebenzylester, Pf. 124.5-126°C.

b) In trifluoracetic acid, we obtain

from this the trifluoracetate of DL-N-(3-alanyl)-3-pheny-1-3-alaninebenzylester.

c) This substance is reacted in methylene chloride/water/sodium bicarbonate with p-amidinobenzoyl chloride and then with benzyl chloroformate to give the starting ester.

Example 35

Similar to example 11, we obtain from benzyl-(S)-3-[[N-[p-[N-[(benzyloxy )carbonyl]amidino]benzoy1]-3-alanyl]amino]-N-[p-[C(benzyloxy)carbonyl]methoxy]phenylethyl]succinamate

51

1' [p-C2-[[N-[N-(p-amidinobenzoyl)-3-alanyl]-L-a-aspartyl]-amino]ethyl]phenoxy]acetic acid in hydrate form (2:7), Pf. 210-213°C.

The starting ester, Pf. 172-174°C, can be prepared as follows:

a) t-butyl-[2-(4-hydroxyphenyl)-ethylcarbamate, benzyl bromacate and potassium carbonate are heated in acetone. t-butyl-[p-[[(benzyloxy)carbonyl]methoxy]phenethyl]c-carbamate, SM: 385 (0.5, M) is obtained.

b) After cleavage of the t-butoxycarbonyl protecting group, it is coupled with the 4-benzyl ester of N-(t-butoxycarbonyl)aspartic acid to give benzyl-(S)-N-[p-[[(benzyloxy)carbonyl methoxy]phenethyl]-3-C1-t-butoxyformamido succinamate, Pf. 178.5-180.5°C.

c) This compound is deprotected in tri-fluoracetic acid and coupled with N-(t-butoxycarbonyl)-(3-alanine) to give t-butyl-[2-[[(S)-2-[(benzyloxy)carbonyl]-1-[Cp-[C(benzyloxy)carbonyl]-methoxy]-phenethyl1]carbamoy1]eth1]carbamoy1]eth1]-carbamate, Pf. 123-124°C.

d) After cleavage with the t-butoxycarbonyl protecting group, it is reacted in methylene chloride/water/sodium bicarbonate with p-amidinobenzoyl chloride and then with benzyl chloroformate to give the starting ester.

Example 36

a) A solution of 280 mg of l-[N-(p-cyanobenzoyl)-3-alanyl]-4-piperidineacetic acid and 1 ml of triethylamine in 15 ml of pyridine is saturated with sufuric acid. After 36 hours, the solution is concentrated and the residue is placed in

52

suspension in ethyl acetate/water. Filtration and drying of the insoluble material gives 255 mg of 1-C N-[p-(thiocarbamoy1)benzoy1]-3-alanyl]-4-piperidinacetic acid.

5 b) A solution of 150 mg of the intermediate product in 15 ml of acetone with 1 ml of methyl iodide is heated for 3 hours at boiling temperature. After cooling the solution to room temperature, a precipitation of 10,130 mg of hydroiodine of l-[N-[p-[l-(methylthio)-formimidoyl]benzoyl]-3-alanyl]-4-piperidinacetic acid (1:1) is observed, Pf. 206-207°C.

c) We maintain 100 mg of l-[N-[p-[l-

(methylthio)formimidoyl]benzoy1]-3-alanyl]-4-piperidin-15 acetic acid and 30 mg of ammonium acetate in 10 mL of methanol for 3 hours at boiling temperature. After cooling to room temperature, the solution is filtered, concentrated, and mixed with diethyl ether. The oil 20 precipitated after decantation of the solvent with water/methanol (10:1) is chromatographed on RP 18 silica gel. The results are obtained.

24 mg 1-[N-(p-amidinobenzoyl)-3-alanyl]-4-piperidinacetic acid iodhydrate (10:1), m.p. 206°C.

The starting nitrile can be obtained as follows:

25 follows:

a) 4.96 g of 4-cyanobenzoyl chloride and 2.67 g of 3-alanine are stirred in 450 mL of 2% sodium bicarbonate solution for 4 hours at room temperature and acidified with concentrated sulfuric acid (pH 6). The solution is concentrated and extracted with ethyl acetate. Drying and concentration of the organic phase yields

"

53

a residue which, when mixed with diisopropyl ether, leads to 4.69 g of N-(p-cyanobenzoyl)-3-alanine, Pf. 155-157°C.

b) Coupling of 635 mg of N-(p-cyanobenzoyl)-

3-alanine with 540 mg of 4-piperidinacetic acid gives, after RP 18 silica gel chromatography with THF/water (85:15) 300 mg of l-[N-(p-cyanobenzoyl)-3-a'lanyl]-4-piperidinacetic acid, SM: 344 (M+H)+.

Example 37

400 mg of t-butyl-4[N-(p-amidinobenzoyl)-3-alanyl]-l-piperazinacetate is stirred in 15 mL of methylene chloride and 15 mL of trifluoracetic acid. The solution is concentrated, the residue is suspended in 5 mL of ethanol, and the undissolved material is separated by filtration. The filtrate is mixed with ethyl acetate, and the residue is filtered through a funnel and dried. Chromatography of the crude product on RP 18 silica gel with water yields 38 mg of trifluoracetate of 4-[N-(p-amidinobenzoyl)-3-alanyl]-l-piperazinacetic acid (5:8), Pf. 157–158°C.

The starting product can be prepared as follows:

a) 2.23 g of N-benzyloxycarbonyl- are coupled

3-Alanine with 2.58 g of piperazine. The residue is suspended in THF, the insoluble matter is separated by filtration, and the filtrate is concentrated. Chromatography of the residue on RP 18 silica gel with water/methanol (2-5%) gives 2.51 g of benzyl-[2-(4-piperazinylcarbonyl)-ethyl]carbamate, SM: 291 (M+.).

54

b) 600 mg of the intermediate product, 0.3 ml of bromacetic acid t-butyl ester, and

25 mg of tetrabutylammonium acid sulfate in 10 ml of toluene and stirred for one hour with 10 ml of 50% sodium hydroxide solution. The organic phase is washed with water and concentrated. The residue is chromatographed on silica gel with ethyl acetate/methanol (9:1), yielding 480 mg of t-butyl-4-[N-[(benzyloxy)carbonyl]-3-alany1]-[-piperaz inacetate, SM: 406 (M+H)+.

c) The intermediate product is hydrogenated in the presence of 200 mg of Pd/C in ethanol for one hour. The catalyst is separated by filtration and the filtrate is concentrated. 290 mg of t-butyl-4-p-alanyl-l-piperazinacetate, SM: 271 (M+), is obtained.

d) The intermediate product and 341 mg of p-amidinobenzoyl chloride are stirred in 20 mL of methylene chloride and 10 mL of saturated sodium bicarbonate. The organic phase is separated, concentrated, and the residue is suspended in ethyl acetate. Filtration through a filter funnel and drying of the crystals yields

400 mg t-butyl-4-CN-(p-amidinobenzoyl)-p-alanyl]-1-piperazinacetate, SM: 418 (M+H)+.

Example 38. Similar to Example 37, 1.6 g of N-[N-[N-(p-amidinobenzoyl)-3-alanyl]-3-(t-butoxycarbonyl)-L-alanyl]-L-valin-t-butylester is deprotected. The crude product is chromatographed on RP 18 silica gel with water/THF (95:5). 867 mg of N-[N-[N-(p-amidinobenzoyl)-3-alanyl]-L-α-αSparty1]-L-valin-t-fluoracetate is obtained, Pf. 162-163°C.

55

The starting ester can be prepared as follows:

a) 4-t-butylester of N-benzyloxycarbonyl-L-aspartic acid and t-butylester hydrochloride of L-valine are coupled to give N-[N-[(benzyloxy)carbonyl]-3-(t-butoxycarbonyl)-L-alanyl]-L-valin-t-butylester, Pf. 75°C (d).

b) The body is deprotected in a similar manner

In example 37c, we obtain N-[3-t-butoxycarbonyl)-L-alanyl]-L-valin-t-butylester, Pf. 71°C.

c) 2.8 g of the intermediate product is coupled with 1.78 g of N-C(benzyloxycarbonyl]-3-alanine and the crude product is chromatographed with ethyl acetate on silica gel. 2.24 g of N-[N-[N-[(benzyloxy)carbonyl]-3-alanyl]-3-(t-butoxycarbonyl)-L-alanyl-L-valin-t-butylester is obtained, Pf. 126°C.

d) Deprotection of 2.2 g of the intermediate product in a manner analogous to example 37c) results in 1.61 g of N-[N-f5-alanyl-3-(t-butoxycarbonyl)-L-alanyl]-L-valin-t-butylester.

e) The reaction of 1 g of p-amidinobenzoyl chloride with 1.61 g of the intermediate product according to the prescriptions of 37d) gives 1.62 g of the starting ester.

Example 39. Similar to Example 37, 1.4 g of N-[N-[N-(p-amidinobenzoyl)-3-alanyl]-3-(t-butoxycarbonyl)-L-alanyl]-3-(p-t-butoxyphenyl)-L-alanin-t-butyl ester is deprotected. The product is suspended in ethanol, the insoluble matter is separated by filtration, and the filtrate is mixed with ether. Filtration through a filtering funnel and

56

the alveolation of the precipitate with 20 ml of isopropanol/ethanol (1:1) gives 801 mg of N-[N-[N-(p-amidinobenzoyl)-3-alanyl]-L-a-aspartyl]-3-(p-hydroxyphenyl)-L-alanin-trifluoracetate (2:5), Pf. 202-204°C (d).

The starting ester can be prepared as follows:

a) N-[(9H-fluoren-9-yloxy)carbonyl]-3-(t-butoxycarbonyl)-L-alanine and 3-(p-t-butoxyphenyl)-L-alanin-t-butylester are coupled to give N-[N-

[(9H-fluoren-9-yloxy)carbonyl]-3-(t-butoxycarbonyl)-L-alanyl]-3-(p-t-butoxyphenyl)-L-alanin-t-butylester.

b) 3.2 g of the intermediate product are stirred in 50 mL of DMP and 5 mL of piperidine, and the reaction solution is concentrated. The residue is suspended in methanol, and the insoluble material is separated by filtration. The filtrate is concentrated and chromatographed with ethyl acetate on silica gel. 2 g of 3-(t-butoxyphenyl)-N-[3-(p-t-butoxy-

carbonyl)-L-alanyl]-L-alanin-t-butylester.

!

c) The intermediate product is coupled with 0.96 g of N-benzyloxycarbonyl-3-alanine and the crude product is chromatographed with ethyl acetate on silica gel. 2.23 g of

N-[N-[N-[(benzyloxy)carbonyl]-3-alanyl]-3-(t-butoxycarbonyl)-L-alany1]-3-(p-t-butoxyphenyl)-L-alanin-t-butylester, MS: 670 (M+H)+.

d) Deprotection of 2.1 g of the intermediate product as in example 37c) gives 1.67 g of N-[N-3_alanyl]-3-(t-butoxycarbonyl)-L-alanyl]-3-(p-t-butoxyphenyl)-L-alanin-t-butylester, SM: 536 (M+H)+.

e) The coupling of 1.5 g of the inter product

57

median with 0.6 g of p-amidinobenzoyl chloride as in example 37d) gives 1.6 g of starting ester, SM: 682 (M+H)+.

Example 40 In a manner analogous to example 1, we prepare N-[N-PN-(5-amidino-2-pyridoyl)-(3-alanyl]-L-a-asparty1]-3-phenyl-L-alanine.

Example A

A compound of formula I can be used in a known manner as an active substance to prepare tablets having the following composition:

Per tablet: active ingredient 200 mg, microcrystalline cellulose 155 mg, maize starch 25 mg, talc 25 mg, hydroxypropyl methylcellulose 20 mg

425 mg

Example B

A compound of formula I can be used in a known manner as an active substance to prepare capsules having the following composition:

Per capsule: active ingredient 100.0 mg, corn starch 20.0 mg, lactose 95.0 mg, talc 4.5 mg, magnesium stearate 0.5 mg

220.0 mg

58

DV4045/12

Claims (1)

DEMANDS 1. Process for preparing acetic acid derivatives of formula ,2 H2N(NH)C-X-Y-CO-Z-CH(Ql)COOQ2 Or Q1 represents a hydrogen, methyl or phenyl group, represents a hydrogen, lower phenyl-alkoyl or a lower alkyl separable under physiological conditions, represents a 1,4-phenylene, 2,5- or 3,6-pyridylene or a 1,4-piperidinylene linked via a carbon atom in position 4 to the Y group, represents a formula group 1. -<CH2>o.2"CONHCH(°3>(CH2Î1-3" <Y } 4, (Y2) •CONHCH2CH(Q)- .(CH2)2NHCOCH2-■NHCO(CH2)3- (Y3) 4 (V) 59 <c^U-Ïh (Y5) Or l (Y?) q3 represents hydrogen, methyl, phenyl, -COOH, -COO-lower alkyl, -CONH(CH2)2-COOH or -CONH(CH2)2"COO-lower alkyl, q4 represents a hydrogen, methyl or phenyl, 5z represents a 1,4-piperazinylene group, a 1,4-piperidinylene group linked via nitrogen atoms in position 1 of the CO group or a group of formula 10 -NHCH(RV or -NHCHtCOR2, 60 r1 represents a lower hydrogen, methyl, phenyl or -COO-alkoyl, 2 r represents the radical of an α-ammo- acid carboxylic acid linked via the amino group, or one of its esters or amides, or a group of formula -NHCH2CH2-Ar, or -CO-R2 represents a carbamoyl group possibly mono- or disubstituted by a lower alkyl group or a pyrrolidinoyl or piperidinoyl group, Ar represents a phenyl or a substituted phenyl by a lower alkyl, lower alkoxy, -COOH, -COO-lower alkyl, -0(CH2)1_4-COOH, 0-CH2)1_4-COOH-lower alkyl, -CONH2, -CONH-lower alkyl, -CON(lower alkyl)2, pyrrolidinyl or piperidinoyl, as well as their physiologically acceptable hydrates or solvation products and salts. characterized in that a) from a compound of formula X'-Y-CO-Z'-CH(Q1)-COO-Q5 II Or Q1 and Y have the meanings given in claim 1, X' is a phenyl group substituted at position 4 by a possibly protected amidino group, or a 4-piperidinyl, and 5 21 or Q has the same meaning as given above. 2 above for Z or Q, in claim 1, Or 61 at least one protecting group is separated, b) in a nitrile of formula Ws c -Q"y" co"2 'C H(a1)-coo-Q 1 (one by transforming the nitrile group into the amidino group and if desired by transforming a compound of formula I into a physiologically acceptable salt or a salt of a compound of formula I into the free acid or base. 2. A process according to claim 1, characterized in that compounds of formula 10 are prepared H2N(HN)C-X-Y3-CONHCH(R11)CH2COOÇ21 i-a where X represents a 1,4-phenylene or a 1,4- piperidinylene linked via the carbon atom in position 4 to the Ya group, 15 y represents a group of formula -<CH2)0.l-c°NHCH<0a)<CH2)i-2- (YlI) 62 (CH2)2NHCOCH2- (Y3) -NHCO(CH2)3- (y4 }gnu51) Qa represents a hydrogen or a phenyl group, r11 represents a hydrogen or -CO-R^ r22 represents the radical of an α-amino acid carboxylic acid linked via the amino group or one of its esters or amides, and q21 represents a hydrogen or a lower alkyl group separable under physiological conditions. as well as physiologically acceptable hydrates or solvation products and salts. 3. Method according to claim 1, where 1 2 X, Z, Q and Q have the meanings given in claim 1, and Y is a group of formula Y1, in particular a group of formula 63 -conh(ch2)2_4-, -ch2conh(ch2)2-, -conhch(c6hg)ch2-, -conhch(conhch2ch2cooh)ch2-, Or -conhch(cooh)ch2--conhch(ch3)ch2-. 4. Method according to claim 1, where 1 2 X, y, Q and Q have the meanings given in claim 1, and Z represents a formula group -nhch2-, -nhch(ch3)-. -nhch(c6h5)-, -nhch(coo-Isobutyl)-, -nhch(co-Va 1)-, -nhch(co-Phe)-. -nhch(co-Tyr)-, -nhch(co-Ser-oc2h5)-. -nhch(co-Leu-o-Isopropyl)-, -nhch(conhch2ch2-cfih4-OCHj)-, -nhch(conhch2ch2-c h4-cooh)-, -nhch(conhch2ch2-c6h4-och2cooh)-, -NHCH(CONH)- or -NHCH(Pyccolidinoyl)- 4» 64 5. A process according to claim 2, characterized in that it prepares compounds from the following group of compounds: N-CN-C4-(p-amidinobenzamido)butyryl]-L-a-aspartyl]-L-valine, N-[N-(p-amidinobenzoyl)-3-alanyl]-3-alanine and in particular N-C N-C N-(p-amidinobenzoyl)-3-alanyl]-L- a-aspartyl]-3-phenyl-L-alanine. 6. A process according to claim 2, characterized in that it prepares compounds from the following group of compounds: N-CN-CN-(l-amidino-4-piperidinylcarbonyl)-3-alanyl j-L-a-asp'artyl ]- 3-phenyl-L-alanine, N-[N-[N-(p-amidinophenylacety1)-3-alanyl]-L-α-aspartyl]-3-phenyl-L-alanine, N-CN-C4-(p-amidinophenylcarbamoyl)-butylyl]-L-a-aspartyl]-3-phenyl^L-alanine, N-CN-C(p-amidinopheny-lcarbaraoyl)acetylj-L-a-aspartyl]-3-phenyl-L-alanine., rac-N-[1-(p-amidinobenzoyl)-3-piperidiny1-carbonyl]-3-alanine, N-C 4-(p-amidinobenzamido)butyryl]-3-alanine, and N-C(DL)-N-(p-amidinobenzoyl)-3-phenyl-3~ alanyl]-3-alanine 7. A process according to claim 1, characterized in that it prepares compounds from the following group of compounds: 65 N— C N—[N-(pramidinobenzoyl)-0-alanyl]-L-a-aspartyl]-L-leucinisopropylester, N-CN-CN-(p-amidinobenzoyl)-0-alanylJ-L-a-aspartyl]-L-valine, N-[N-[N-(p-amidinobenzoyl)-0-alanylJ-L-a-aspartyl]-3-(p-hydroxyphenyl)-L-alanine, N-[N-[5-(p-amidinobenzamido)valéry1]-L-a-aspartyl]-3-phenyl-L-alanine, N-[5-(p-amidinobënzamido)valéry1]-L-a-aspartic acid, isobutylester, N-CN-CN-(p-amidinobenzoyl)-0-alanyl]-L-a-aspartyl3-L-serineethyl ester and N-C N-[C(R)-l-(p-amidinobenzoyl)-3-pyrrolidinyl]■ carbonyl]-L-a-aspartyl]-3-pheny1-L-alanine. 8. Method according to claim 1, characterized in that compounds from the following group of compounds are prepared: N,N'-[[(S) — (p-amidinobenzamido)ethylen] dicarbonyl]di-0-alanine, N2-(p-amidinobenzoyly-N4-(2-carboxyethyl)-L-asparagine, N-C 5-(p-amidinobenzamido)valeryl]-0- alanine, rac-N-C [1- [3 - (l-amidincJ-4-piperidinyl) propionyl]-3-piperidiny1Jcarbonyl]-0-alanine, N-[C(S)-l-(p-amidinobenzoyl)-2-pyrrolidinyl]-acetylJ-0-alanine, (S)-3-C[N-(p-amidinobenzoyl)-0-alanyl]-amino-3-C(p-methoxyphenethyl)carbamoyl]propionic acid, N-C C(R)-l-(p-amidinobenzoyl)-3-pyrrolidinyl]-carbonyl]-0-alanine, 66 α-amide of N-[N-(p-amidinobenzoyl)- 2-methy1-3-alany1]-L-aspartic acid, N-[N-[(p-amidinophenyl)acetyl]-3-alanyl]- 3-alanine, 5 rac-N-[[[1-(p-amidinophenyl)acetyl]-3-piped- ridinyl]-carbonyl]-3-alaninebenzylester, rac-N-[Cl-[(p-amidinophenyl)acetyl]-3-piperidinyl]-carbonyl]-3-alanine/ N-[N-[N-[3-(l-amidino-4-piperidinyl)propio-10nyl]-3-alanyl]-L-a-aspartyl]-3-phenyl-L-alanine/ (S)-3-[[DL-N-(p-amidinobenzoyl)-3-methyl1-3-alany1]amino]-y-oxo-l-pyrrolidinebutyric acid, DL-N-[N-(p-amidinobenzoyl)-3~alanyl]-3-methyl-3-alaninef 15 N-CDL-N-(p-amidinobenzoyl)-2-phenyl-3~ alanyl]-3-alanine, DL-N-C N-(p-amidinobenzoyl)-3-alanyl1-2-methyl-3~alanine/ DL-N-CN-(p-amidinobenzoyl)-3~alanyl]-2-20 phenyl-3-alanine, Acid p— C 2— C[N-[N-(p-amidinobenzoyl)-3~ alany 1 ] -L-a-aspartyl ]amino ] ethy.l ]benzoic, DL-N-CN-(p-amidinobenzoyl)-3-alanyl-3-phenyl- 3-alanine, 25 Acid [p-C2-[[N-[N-(p-amidinobenzoyl)-3- alany1]-L-a-asparty1]amino]ethyl]phenoxy]acetic. Acid 1-[N-(p-amidinobenzoyl)-3-alanyl]- 4-piperidinacetic acid, 4-[N-(p-amidinobenzoyl)-3~alanyl]-30 l-piperazinacetic acid. 1 67 9. A process for preparing pharmaceutical preparations, in particular for the treatment or prophylaxis of diseases caused by the binding of adhesive proteins to blood platelets, as well as by the agglutination of blood platelets and cell-cell binding. characterized in that a compound is placed "in a galenic administration form," according to claim 1 or 2. 10. Application of a compound according to claim 1 or 2 to the preparation of medicinal products for the treatment or prophylaxis of diseases which are caused by the binding of adhesive proteins to platelets as well as by the agglutination of blood platelets and cell-cell adhesion, in particular for the treatment or prophylaxis of platelet clots, thrombosis, strokes, myocardial infarction, of inflammation or arteriosclerosis, or as an antitumor agent or as an agent for wound healing. 11. Acetic acid derivatives of formula X'-Y-CO-Z-CHtQ^-COOQ2 n» where 1 X' is a phenyl group substituted at position 4 by a possibly protected amidino group or a 4-piperidinyl and 1 2 Y, Z, Q and Q have the meanings given in claim 1, where the molecule contains at least one easily separable ester group or one protected amidino group.