PT101001A - NEW UTEIS TRIPEPTIDE DERIVATIVES AS ANTI-INFLAMMATORY AGENTS - Google Patents

Description

6 ^ 5432 Case 4514

1 NEW USEFUL TRIPEPTIDE DERIVATIVES AS INFLAMMATORY AGENTS

ORIGINS OF THE INVENTION 5 10 15

Mod. 71-20,000 ex. Certain types of tripeptide derivatives with various biological activities are known. The following references describe such tripeptide derivatives: U.S. Patent 4,242,329, to Claeson, Simonasson & Arielly of 30 December 1980; U.S. Patent 4,316,889 to Bajusz, Hasenohrl, Barabas & Bagdy of 23 February 1982; U.S. Patent 4 399 065, from Bajusz, Hasenohrl, Darabas & Bagdy, 16 August 1983; US Patent 4,401,594 to Umezawa, Takeuchi, Aoyagi, Ishii, Saino & Someno of 30 August 1983; U.S. Patent 4,478,745, Bajusz, Hasenohrl, Barabas & Bagdy of 23 October 1984; U.S. Patent 4,528,133, to Kasafirek, Fric, Slaby & Robalova of 9 July 1985; U.S. Patent 4 596 789, to Dutta, Stein, Trainor & Wildonger, 24 June 1986; U.S. Patent 4,623,639. of Hassall. Johnson & Roberts, judgment of 18 November 1986; US Patent 4,703,036 to Bajusz, Hasenohrl, Bagdy, Barabas, Sioszegi, Fittler, Jozsa, Horvath & Jozst of 27 October 1987; U.S. Patent 4,880,780, to Trainor & Stein, 14 November 1989; U.S. Patent 4,883,863, to Abe, Yaginuma, Nagasawa & kuroiwa of 28 November 1989; U.S. Patent 4,902,781, to Mizoue, Okazaki, Hanada, Omura & Amamoto of 20 February 1990; PCT Patent Application 84/00365, from Galpin & Wilby, 2 February 1984; Japanese Patent Application 47-30618 to Toray Inds. Inc., November 9, 1972; Japanese Patent Application 57-145846 to Mitsubishi Chem. Ind KK of 19 July 1987; Japc Patent Application > No. 58-164563 to Daiichi Kagaku Yaku, 25 March 1982; Aoyagi, Miyata, Nanbo, Kojima, Matsuka, Takeuchi, & Umezawa, " Biological Activities of Leupeptins " [J " Biological Activities of Leupeptin " J, The Journal of Antibiotics 3 35 1 5 10 15

Mod. 71-20,000 ex. - 90 (08 20 25 Ô5432 Case 4514

| _ Antibiotic News _ | , Vol. XXII, No. 11 (Nov. 1969), pp. 558-568; Bajusz, Barabas, Tolnay, Szell & Bagdy, " Inhibition, od Thrombin and Trypsin by Tripeptide Aldehydes " | _ Inhibition of Thrombin and Trypsin through Tripeptide Aldehydes " _ | , Int J, Peptide Protein REs., Vol. 12 (1978), pp. 217-221; Gaal, Bacsy & Rappay " Cationic Ferritin Uptake by Cultured Anterior Pituitary Cells Treated with the Proteinase Inhibitor, BOC-DPhe-Phe-Lys-H " | _ " Purified Proteinase, BOC-DPhe-Phe-Lys-H ", Histochemistry, Vol. 88 (1988), p. 401-406; Makara, RAppay, Garamvology, Nagy, Danko & Bajusz, " The Tripeptide Aldehyde, Boc-DPhe-Phe 2+ i -lysinal, is a Novel Ca Cannel Inhibitor in Pituitary Cells " Tripeptide Aldehyde, boc-DPhe-Phe-Lysinal, is a novel inhibitor of novel Ca-channel in pituitary cells ", European Journal of Pharmacology, " European Journal of Pharmacy " J, Vol 151 (1988), pp. 147-149; Nagy, Makara, Horvath, Rappay, Kurcz & Bajusz, " Tripeptide Aldehyde Protease Inhibitors May Depress in Vitro Prolaction and Growth Hormone Release " | _ " Tripeptide Aldehyde Protease inhibitors may depress in vitro the release of Prolactin and growth hormones " J, Endocrinology, Vol. 116, No. 4, (1985), pp. 1426-1432; Rappay, Makara, Bajusz & Nagy, " Various Proteianse Inhibitorf Decrease Prolaction and Growth Hormone Release by Previous Pituitary Cells " Several Proteinase Inhibitors Decrease Prolactin and Growth Hormone Release Through Pituitary Cells, " Life Sciences " Life Sciences ", Vol. 36 (1985), pp. 549-555. Some of these references disclose tryptophan tripropion derivatives having anti-inflammatory activity. It is an object of this invention to provide novel tripeptide derivatives useful as anti-inflammatory agents. It is a further object of this invention to provide derivatives of three novel peptides which reduce bone resorption and / or heterotopic bone formation associated with rheumatoid arthritis. 4 30 65432 Case 4514

It is a further object of this invention to provide novel pharmaceutical compositions comprising certain types of tripeptide derivatives. It is a further object of this invention to provide methods for treating diseases characterized by inflammation.

SUMMARY OF THE INVENTION

This invention involves anti-inflammatory compounds having the following structure:

X- (CH9) -V-Z-CH-C (O) -NH-CH-C (O) -NH-CH-C (O) -Y

Z 'H wherein 15

Mod. 71 -20,000 ex. (A) is an integer from 0 to about 2; (b) -R is selected from straight or branched chain alkyl, saturated or unsaturated with 1 or 2 double bonds, having 1 to 6 carbon atoms; and cyclic alkyl, whether or not saturated with 1 or 2 double bonds, having 2 to 13 carbon atoms, and the carbon atom to which it is attached R 1 is in the D or L configuration. (c) -R 1 is selected from branched chain alkyl, saturated or unsaturated with 1 or 2 double bonds, having from 3 to 6 carbon atoms, cyclic alkyl, saturated or not with 1 or 2 double bonds, having 3 to 13 carbon atoms; and arylalkyl, the alkyl portion being saturated and having 1 to 13 carbon atoms; the carbon atom to which -R 1 is attached in the L configuration; (d) -R " â € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒ (CH9) -A-NEL or - (CH9) -A-B- A-is a covalent bond, p-phenyl or p-cyclohexyl, and -B- is a covalent or -NH- bond; the carbon atom to which -R " is connected in L configuration; 5 35 5 10 15

Mod. 71-20,000 ex. - 90/08 25

65432 Case 4514 (e) -Y is hydrogen or trifluoromethyl; (f) -Z- is -O- or -NH-; (g) -V- is selected from -OC (O) -, -N (Q) C (O) -, N (Q) C (S) -, -C (O) (O) (OH), when -V is -O (O), -Z- is -NH-; (h) X is selected from branched chain cyclic alkyl having at least 2 branching and aryl, each having 5 to 20 carbon atoms; and (i) Q is selected from hydrogen; and straight or branched chain alkyl, whether or not saturated with 1 or 2 ligands " double bonds having 1 to 6 carbon atoms; or -Q and -X are covalently linked, forming with the cyclic moiety which includes the nitrogen to which the -Q is attached and between 5 and 20 carbon atoms. The subject of the invention also includes pharmaceutical compositions comprising the above-mentioned compositions and methods for the treatment of inflammation or pain by use of such compounds and compositions.

DETAILED DESCRIPTION OF THE INVENTION

As used herein, the term " alkyl ", unless otherwise specified, refers to carbon-containing chains, which may be linear, branched or cyclic; which can be saturated or not; and which can be replaced or not. As used herein, the term " cyclic alkyl " may have all or only a portion of the total number of carbon atoms indicated as being on the alkyl group on the cyclic ring itself includes monocycloalkyl, bicycloalkyl and / or tricycloalkyl. The ideal alkyl is that referred to in this paragraph, unless stated otherwise. The ideal alkyl is saturated. 6 35 1 i 5 10 15

80 / 06-> W-65-W-W-65432 Case 4514 The ideal alkyl is unsubstituted. Suitable alkyl substituents are selected from halo, amino, hydroxy, alkoxy, C ano,,, nitro, aryl and trifluoromethyl. as used herein, " alkoxy " is -O-. The alkyl is mono-, di- or trisubstituted, in particular monosubstituted. As used herein, the term " aryl ", unless otherwise specified, means aryl rings, which may be unsubstituted or substituted. The aryl is ideal, as referred to in this paragraph, unless other examples are given. The ideal aryl is phenyl and naphthyl, in particular phenyl. Preferred is mono-, di-, trianyl or unsubstituted; more preferred are monosubstituted or unsubstituted aryl, especially unsubstituted aryl. Suitable aryl substituents include alkyl, halo, amino, hydroxy, cyano, nitro and trifluoromethyl. The term " alkyl " as used herein denotes alkyl substituted by aryl. Arylmethyl is a preferred alkyl group. Here are the most used abbreviations here. the abbreviations for amino acids may refer to the amino acid itself, or more often an amino acid moiety may refer to the amino acid itself, or more often an amino acid moiety, which is part of a peptide or a peptide structure.

Amido-acid Protecting Group Arginyl Cyclic arginyl Leucilo Isoleucyl Lysyl Penylalanyl p-guanidinophenylalanyl 35 Prolyl AA PG Arg cArg Leu Ile LYS Phe p-Gphe Pro 7 1 10

Morph. 71-20,000 ex. -90/08 20 25 65432 Case 4514

Seryl Threonyl allo-Threonyl Triptophyl Valyl 1- Naftilaianyl 2- Naftilalanyl

Ser Thr aThr Trp Val-1-Nal-2-Nal

Na 1 (collectively)

Ciclohexilalanila t-Butilalanila 1- 2- Adamantilalanlla Adamantilalanila Omithilo Adamantilglicilo t-aminocyclohexyl Butiglicilo Aminociclopentilo Ciclohexilglicllo Homofenllalanila p-Chlorofenilalanlla Nitrofenilalanila p-t-butyloxycarbonyl Tienilalanila Carbobenzyloxy adamantyloxycarbonyl Benzyl Adamantoilo Adamantaneacetilo Mentlloxicarbonllo Homoadamantiloxicarbonilo Fechiloxlcarbonilo Isomentiloxicarbonilo Isipinocamfaniloxicarbonilo endo-3,5-naphthyloxycarbonyl Borniloxicarbonilo Dimetiladamantiloxicarbonilo

Cha t-Bal 1-Adl 1 2-Adl

Orn Adg t-Bug Ach Acp Chg Hph p-Cph p-Nph • Adi (collectively)

Xhi Cbz or Z Boc Adoc Bn

Ad Ada Moc Sadoc Foc Imoc Ipoc eBroc Noc 3,5-Dmadoc 8 35 í 6 * 5432 Case 4514

1 5 10 15

Mod-71 · 20,000 ex. - 90/08 20 25

Morfilinoilo Mnc Fluorenylmethoxycarbonyl Fmoc Noradamantoilo Norad Adamantilaminocarbonilo Adac triphenylmethyl Rt isopropyl alcohol IPA Cinanofosfato DECP Trifluoroacetate TFA potassium Hexametilodisilazido KHMDS Hidrio lithium aluminum LAH p-toluenesulfonic acid pTSA Tetrahydrofuran THF Dimetilossulfóxido DMSO Acetyl Ac Methyl Me Ethyl Et 3-chloroperoxybenzoic MCPBA dimethylformamide DMF TEA triethylamine hydroxide diissobutilalumínio DIBAL-H Silane Trifluoromethylthiomethyl cf3tms trihydrate fluoride tetra-n-butylammonium Rapid atom bombardment- TBAF mass scopia spectrum FAB-MS

The novel anti-inflammatory compounds relating to the subject matter of this invention are those having the following chemical structure: R R 'R " X- (CH9) -V-Z-CH-C (O) -NH-CH-C (O) -NH-CH-C (O) -Y (1)

In structure (1), R is selected from the group consisting of straight or branched chain alkyl, saturated or unsaturated with 1 or 2 double bonds, having 1 to 6 carbon atoms; and cyclic alkyl, saturated or unsaturated with 1 or 2 double bonds, having 3 to 13 carbon atoms. 0 -R ideal is 9 30 65432 Case 4514

saturated alkyl. The ideal alkyl is unsubstituted or arylalkyl, especially benzyl or naphthal. The ideal cyclic alkyl is cyclic Cg-Cg (preferably C -C-C)) in methyl or ad-10-15.

Mod. 71-20,000 ex. The ideal R is hydrophobic, preferably with the hydrophobic concentration near the carbon atom to which it is attached -R. Specific examples of suitable -R include t-butyl, 1,1-dimethylpropyl, i-butyl, s-butyl, neo-pentyl, cyclohexyl, cyclohexylmethyl, adamantyl, naphthal; preferably -R should be t-butyl. In structure (1), the carbon to which the -R is attached is in a D or L configuration, preferably D. The structure -Z-CH (R) -C (O) is an amino acid moiety in dian designated as -AA-), when -Z- is -NH; the amino-acid moieties ideal for -AA- include t-Bug, Val, Ile, Leu, Chg, Cha, Phe, Nal, Trp and Adg; preferably t-Bug, Val and Ile; 1 and more preferably, -AA- must be T-Bug. In structure (1), -R 'is selected from branched chain alkyl, saturated or otherwise, having 1 or 2 double bonds, having from 3 to 6 carbon atoms; cyclic alkyl, saturated or unsaturated with 1 or 2 double bonds, having 3 to 13 carbon atoms; and arylalkyl, the alkyl portion being saturated and having 1 to 3 carbon atoms. The branched or cyclic alkyl should be saturated. Suitable branched alkyl has from 3 to 5 carbon atoms; preferably the branched alkyl should be i-butyl. The ideal cyclic alkyl should have a Cg-Cy, preferably Cg-Cg, cyclic ring linked to methylene, ethylene or n-propylene (preferably methylene or ethylene, more preferably methylene), which is attached to the carbon atom in structure (1), to which the -R 'is attached. Suitable arylalkyls are benzyl, p-hydroxybenzyl and naphthal. The most suitable R 'is selected from i-propyl, i-butyl, s-butyl, cyclohexymethyl, benzyl and naphthal; preferably benzyl. In structure (1), the carbon atom to which the -R 'is attached has a L-configuration. The structure -NH-CH (R') - 10 30 f 6'5432 Case 4514

1 5 10 15 Μοβ. 71-20,000 ex. - C (O) - is an amino acid moiety (hereinafter referred to as -AA3 -); the amino acid moieties which are ideal for -AA2-2 include Phe, Nal, Cha, Leu and Ile; more preferably -AA-must be Phe. In structure (1), -R " â € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒ- (CH2) cyclohexyl, and -B- is a covalent bond or -NH- When -A- is a covalent bond, m must be 2-5, preferably 2-4, more preferably 3 or 4. When -A p-phenyl or p-cyclohexyl, m must be 1-4, preferably 1-3, more preferably 1. -A- must be a covalent bond. -B- must be -NH- Preferably, -R " In the structure (1), the carbon to which the -R "is attached is in the L-configuration the structure -NH- ( CH (R ') -C (O) is an amino acid moiety (hereinafter designated as AA-); the amino acid moieties ideal for AA include 3 Arg, Lys and p-Gphe; Preferably-AA- should be Arg. In structure (1), -Y is hydrogen or trifluoromethyl. In structure (1), -Z- is -O- or -NH-, -Z- must be -NH-.

In structure (1), -V- is selected from -OC (O) -, -N (Q) C (O) -, -N (Q) C (S), -C (o) - and -P (O) (OH). -V- must be -OC (O) or -N (Q) C (O). Preferably -V- should be -OC (O) -. When -V- is -OC (O) -z- is -NH-.

In structure (1), -X- is selected from C1-6 alkyl, branched chain alkyl having at least 2 branches and aryl, each containing 5 to 20 carbon atoms. -X must have 5 to 15 carbon atoms; preferably -X has 8 to 12 carbon atoms. The ideal alkyl moieties of -X are saturated. -X is preferably unsubstituted or substituted by unsubstituted alkyl or aryl. Suitable cyclic alkyls are monocycloalkyl, bicycloalkyl,

Mod. 71-20,000 ex. And tricycloalkyl, preferably bicycloalkyl and tricycloalkyl, in particular tricycloalkyl. The ideal cycloalkyl has 5 or 6 carbon atoms, preferably 6 carbon atoms, in each cyclic ring: A suitable radical is adamantyl, -X aryl being naphthyl and phenyl, unsubstituted or substituted by alkyl. Particularly preferred aryls include: .naphthyl and fluorenyl. When -X is aryl or cyclic alkyl, n is 1. In structure (1) when -V- is -N (Q) C (O) -or-N (Q) C (S) -Q is selected from hydrogen : straight or branched chain alkyl, saturated or unsaturated with 1 or 2 double bonds, having 1 to 6 carbon atoms, or -Q- and -X are covalently linked to form a cyclic moiety which includes nitrogen Preferred alkyl groups are unsubstituted or substituted by unsubstituted alkyl or unsubstituted aryl. The compound of formula (I) wherein X is alkyl, (hereinafter referred to as the "X") is preferably monocyclic, bicyclic or tricyclic, preferably monocyclic, preferably Boc , Adoc, Fmoc, Ada., Adac; Mococ, Norad, Hadoe, Foc, Imoc, Ipoc, 3,5-Dadoz, eBroc, Noc and Moc; X 'should be Adoc, Ipoc and eBroc, preferably Adoc. anti-inflammatory The ideals of this invention include pharmaceutically acceptable salts of the compounds recited above. The ideal salts include addition salts formed between strong acid and the above compounds. Particularly preferred are salts which include addition salts wherein -R " is per-toned, resulting in a positive charge in the -R " half. Preferred compounds of this invention are often associated with 1 or more water molecules in the form of hydrates. The ideal anti-inflammatory compounds of this invention include the following compounds:

Mod. 71-20,000 ex. - 90/08 20 25 30 65432 Case 4514

Boc-D-Phe-Phe-Arg-H Boc-D-Cha-Arg-H Boc-D-Phe-Arg-H Boc-D-Nal- Boc-D-Phe-Hph-Arg-H Boc-D-Hph-Phe-Arg-H Boc-D-Cha-Phe-Arg-H Boc-D-Hph-Phe-Arg-H Boc-D-Phe-Phe-Arg-H 2-Arg-H-Ad-D-2-Nal-Arg-H Adoc-D-2-Nal-2-Arg- H Ad-D-Phe-Arg-H Foc-Val-Phe-arg-H Adoc-D-Hph-Phe-Arg-H Boc-D-Trp-Phe-arg-H Boc-D-2-Adl-Phe-Arg Boc-D-1-Adl-Phe-Arg-H Boc-D-Phe-Arg-H Foc-D-Phe-Phe-Arg-H -H Adoc-D-Bug-Phe-Arg-H Ad-D-Phe-Phe-Arg-CF3 Adoc-P-Val, Phe-Arg-H Adoc-Dt-Bug-2-Nal- Pt-Bug-Phe-Arg-H Adoc-D-Chg-Phe-arg-H Adoc-D-tBug-p-Cph-Arg-H Adoc- Phe-Arg-H Adoc-Ach-Ph-Arg-H Adoc-Phe-Arg-H Adoc-D-ADg-Phe-Arg-H Adoc-D-Orn-Phe-Arg-H 13 35 1 5 10 15

Mod. 71 -20,000 ex. - 90/08 20 25 65432 Case 4514

Adoc-D-Lys-Phe-Arg-H Adac-DT-Bug-Phe-Arg-H (-) - Moc-D-Leu-Phe- Dt-Bug-Phe-Arg-H Norad-Dt-bug-Phe-Arg-H Norad-Dt-Bug-Phe-Arg-H Adoc-Dt-Bug-Phe-Lys, H Hadoc- Arg-H Adoc-Dt-Bug-Cha-Arg-H Adoc-Dt-Bug-Leu-Arg-H Foc-Dt-Bug-Phe-Arg-H Imoc-Dt-Bug-Phe-Arg-H (-) -Ipoc-Dt-Bug-Phe-Arg-H 3,5-Dmadoc-Dt-bug-Phe-Arg-H (+) - Ipoc-Dt-Bug-Phe- Bug-Phe-Arg-H Adoc-Dt-Bug-Phe-Arg-CF-Adoc-Dt-Bug-Phe-p Boc-D-Cha-p-Gphe-CF-Adoc-D-Val-Fhe-p-Gphe-CFg Boc-D-Phe- 2-Nal-Gphe-CF 3 Adac-D-Bug-Arg-CF 3 MNC-D-Phe-2-Nal-Arg-CF 3 The amino acids have a L-configuration unless indicated as D in the above list .

PHARMACEUTICAL COMPOSITIONS

The pharmaceutical compositions of this invention comprise a safe and effective amount of a tripe derivative

Mod. 71 - 20,000 ex. 90/08 20 25 65432 Case 4514

as defined above and a pharmaceutically acceptable carrier. Such compositions generally comprise about 0.1% to about 95% by weight of a tripeptide derivative, preferably from about 1% to about 90%, more preferably from about 5% to about 75%. As used herein, the term " pharmaceutically acceptable carrier " refers to 1 or more compatible solid or liquid fill diluents or encapsulating substances which are suitable for administration to humans or other animals. The term " compatible " as used herein denotes the components of the pharmaceutical compositions capable of being admixed with the tripeptide derived from this invention and with each other so that there is no action that reduces substantially the pharmaceutical efficacy of the composition in normal use situations. Pharmaceutically acceptable carriers must of course have a sufficiently high purity and a sufficiently low degree of toxicity that make them suitable for administration to humans or animals to be treated. Some examples of substances that serve as pharmaceutically acceptable carriers are sugars, such as lactose, glucose and sucrose; starches such as corn starch and potato starch, cellulose and derivatives, such as sodium carboxymethylcellulose, ethylcellulose and cellulose acetate; alca powdered strips; malt; gelatins; talc, stearic acid; magnesium stearate; calcium sulfate; vegetable oils, such as peanut oil, cottonseed oil, sesame oil, corn oil, and theobroma oil; pollutants, such as propylene glycol, glycerin, sorbitol, mannitol, and polyethylene glycol, alginic acid, pyrogen-free water; saline in isotonic, phosphate buffer solutions; cocoa butter (suppository base); emulsifiers, such as water-wetting agents and lubricants, such as sodium lauryl sulfate, colorants, flavorings, stabilizers, antioxidants, preservatives, as well as other comparable substances.

5 10 15

Mod. 71-20,000 ex. - Non-toxic chemicals used in the pharmaceutical field. Other pharmaceutically acceptable pharmaceutical acceptable pharmaceutical additives may be included for use in the compositions of this invention. The choice of a pharmaceutically acceptable carrier to be used in conjunction with the tripeptides derived from this invention is determined by the manner in which the active is to be administered. The ideal modes for administering the assets of this invention are injection, ingestion, inhalation and topical application. The pharmaceutically acceptable carrier together with the tripeptide derived from this invention is used in a concentration sufficient to provide a convenient size to enable a dosing. The pharmaceutically acceptable carrier, in its entirety, generally comprises from about 5% to about 99.9% by weight of the pharmaceutical compositions of this invention, preferably from about 10% to about 99%, more preferably from about 25% and about 95%.

The total individual dosages of the tripeptides derived from this invention in the pharmaceutical compositions are usually from about 1 mg to about 1000 mg, preferably from about 50 mg to about 800 mg, more preferably from about 100 mg to about 500 mg .

METHODS FOR PRODUCTION OF ANTI-INFLAMMATORY ACTIVITY

AND ANALGESICS

This invention further comprises methods for the production of anti-inflammatory and / or analgesic activity in humans or animals by administering (in a non-treatment) a safe or effective amount of a tripeptide derivative of this invention . The amount may be given in an individual dose or in repeated multiple doses during the course of treatment. Although doses greater than those described herein are effective in reducing inflation and producing analgesia,

1 5 10 16

Mod. 71-20,000 e *. - 90/08 20 25 guns to avoid adverse side effects. The tripeptide derivatives and compositions of this invention can be used to reduce inflammation in various disorders in the deeper structures, muscles, tendons, vesicles and irritations, associated with diseases and traumas, for treatment and prevention of pain. The optimal modes for administration of the tripeptide derivatives of this invention are injection, ingestion, inhalation and topical application. Specific modes of administration include, but are not limited to, oral ingestion, injection (such as intramuscular, intravenous, intraperitoneal, intradermal and subcutaneous); inhalation and topical application, such as at the transdermal, oral, muscular and sublingual levels. As used herein, the term " safe & effective amount " means an amount of a sufficiently high tripeptide derivative or composition to modify the condition to be treated in a positive and significant manner but low enough to avoid serious side effects (at a reasonable benefit / risk ratio) under medical supervision. A safe and effective amount of a tripeptide derivative or a tripeptide composition will vary depending on the particular condition being treated, the age and physical condition of the patient, the severity of the condition, the duration of the treatment, the nature of the concurrent therapy, the active used, the pharmaceutical carrier used, and related factors that are of the doctor's responsibility. The ideal daily doses of tripeptide derivatives of this invention are from about 0.1 mg / kg of body weight to about 500 mg / kg of body weight, preferably from about 1 mg / kg to about 100 mg / kg, more preferably between about 2 mg / kg and about 50 mg / kg. One to about 4 simple dosages can be administered per day, ie, preferably about 2 to about 3 simple doses per day. The ideal amounts of the tripeptide derivatives administered by injection are between about 0.1 mg / kg / day and about 30 mg / kg / day.

* 65432 Case 4514 at 50 mg / kg / day, preferably from about 1 mg / kg / day to about 10 mg / kg / day. The ideal amounts of orally administered tripeptide derivatives are from about 1 mg / kg / day to about 500 mg / kg / day, preferably from about 5 mg / kg / day to about 100 mg / kg / day . The ideal amounts of the tripeptide derivatives administered by inhalation are between about 0.1 mg / kg / day and about 500 mg / kg / day, preferably between about 5 mg / kg / day and about 100 mg / kg /day. Preferred amounts of the topically administered tripeptide derivatives are from about 1 mg / kg / day to about 500 mg / kg / day, preferably from about 50 mg / kg / day to about 250 mg / kg / day.

METHODS FOR SYNTHESIS OF COMPOUNDS

The following non-limiting schemes and examples demonstrate the methods of synthesizing tripeptide derivatives of this invention.

Peptide aldehydes are synthesized according to schemes A and B, which follow. In Scheme A, the Z-protected arginine derivative 1 is cyclized with the use of isobutyl chloroformate and an appropriate base in an inert solvent and deblocked with anhydrous HCl to provide the cyclic arginine derivative 2: (Z) -Arg. HCL. This compound can be added to a dipeptide moiety having a terminal amino protecting group using diethyl cyanophosphate and an appropriate amino base at room temperature to form a tripeptide nucleus. The ester 3 can be effectively reduced with lithium aluminum hydride in THF (Z) removed by catalytic hydrogenolysis for the production of a target peptide 4. In a similar sequence of reactions, the cyclic arginine derivative 2 is coupled to a Boc-protected amino acid under conditions similar to those used for the formation of 3 to form a dipeptide unit 5. The Boc group is then removed with anhydrous HCl in dioxane and the salt 18

The resulting amine is bound to another amino acid derivative under the same conditions after exposure to the base. This, again, forms the tripeptide nucleus 3. As before, this compound is converted to 4.

SCHEME A

In Scheme B, the tripeptide ester 6 which is synthesized using standard peptide chemistry is reduced to alcohol with CaBH 4, at 0 DEG in THF and, subsequently, oxidized with DMSO and oxalyl chloride to give the protected peptide aldehyde. Removal of the protecting group by catalytic hydrogenolysis provides the target peptide aldehyde.

Scheme B Z 1) CaBH 4

PG-AA-AA-Lys-OMe-PG-AA-AA-Lys_H (2) oxidation] 3) H2 / Pd-Black

Trifluoromethyl ketones; are synthesized according to 19 '65432 Case 4514

with schemes C and D that follow. In Scheme C, diamine 8 is monoprotected as the Boc derivative and the resulting free amine is oxidized with hot 3-chloroperoxybenzoic acid in dichloroethylene to give the nitro derivative 9. This compound is condensed with hydrate CF3 CHO in THF to provide a nitro alcohol and the reduced nitro to the amine to give 10. The subsequent reaction of this free amine with a dipeptide moiety in the presence of diethyl cyanophosphonate and a suitable amine is followed by selective removal of Boc with anhydrous guanylacaric acid from the resulting amine free with 3,5-dimethylpyrazole-carboxamidine nitrate in dry DMF and oxidation with Dess-martin periodinane (cf. Dess, DB & JC Martin, " Rapid access oxidizer 12-1-5 for primary and secondary alcohols for aldehydes and keto-nas "(Readily Accessible 12-1-5 Oxidant for the Conversion of Primary and Secondary Alchools to Aldehydes and Ketones Journal of Organic Chemistry, Vol.48 (1983), pp. 4155-4156) to give ketone trifluoromethyl 12

CH 2 N (XCH 2). n = 1-3 8 m2 «-is? Boc-ffl / N (CH 2) n /

1) K CO CF, CHO

(CF-1) H, 1) H + 2) guanflate 3) oxidation NHBoc 10 PG-AA-AA-HN-pear;

| (3H, s), 3.35 (3H, s). (ch2), | 11 1 5 10 15

Mod. 71-20,000 e *. 20 25 §5432 Case 4514

Other trifluoromethyl ketones are synthesized by a different method. In Scheme D, the ester 13 is tritylated in dichloromethane with trityl chloride and TEA and the ester is reduced with DIBAL-H in THF and oxidized to the aldehyde 14 under conditions identical to those used in the conversion of 6 to 7. This compound is subjected the reaction with CF 3 TMS (cf. Krishnamurti, R., DR Bellew & BKS Prakash, " Preparation of Trifluoromethyl and other Perfluoroalkyl Compounds with (Perfluoroalkyl) trimethylsilanes " Preparation of Trifluoromethyl and Other Perfluoroalkyl Compounds with Perfluoroalkyl) Trime-thyl Journal of Organic Chemistry, Vol.56 (1991), pp.948-989) and tetra-n-butylammonium fluoride trihydrate in THF, the product being reduced with anhydrous tin dichloride to give the min. The selective coupling of a dipeptide unit is effected at a lower amino nucleophilic location with diethyl dialkophosphonate and a suitable base, the anilinofin is guanylated with N, N'-bis-Cbz-s-methyl, isothiourea (Example 74 and a catalysis The oxidation of the alcohol with Dess-martin periodinane and the removal of Z protecting groups through the hydrogenolysis with black palladium on DMF gives the trifluoromethyl ketone 17. 21 30 65 432 Case 4514

10 15

Mod. 71-20,000 ex.- 90/08 20

SCHEME D

ELN 'VC09Me 1 aq: Eet-2-yl): DIBAL-H 3) oxidation 13

1) CF1 TMS / TBAF - (2) SnCl

1) PG-M-M-CH-H-N'-CF-PG-AA-AA-HNY-CF3

2) guanylate 15 IQ1 Ψ 2) Ho

'NH 4 NHZ 16

N- (Cbz) amidino-2- (Boc) aminovalerolactate-Boc, -Cbz-arginine (5.0 g, 9.84 mmol) is cooled to 0 ° C in 150 mL of THF in an inert atmosphere. To this solution is added dropwise 1.5 ml (10.82 mmol) of the following is the dropwise addition of 1.4 ml (10.82 mmol) of isobutyl chloroformate . After 0.5 hours at 5 ° C, the coolant was removed, the mixture stirred at room temperature for 4 hours. The reaction mixture is poured into a mixture of ethyl acetate (100 ml) and saturated sodium bicarbonate, the phases being separated into a separatory funnel. The organic phase is washed with saturated sodium chloride, dried over magnesium sulfate and filtered, and the solvent is removed. The residue is chromatographed on silica gel to provide 4.0 g of product. Recrystallization from acetone affords 22 35 10 15

Mod. 71-20,000 ex. - 90/08 20 25 30 * 65432 Case 4514

/ pure product analytically. = 0.55 (5% w / CH 3 Cl 3). Boc-D-Phe-Phe-OBn: is added to a mixture of Boc-D-Phe-OH (1.00 g, 3.77 mmol) and Phe-OBn.p-TSA -in salt (1.61 g, 3.77 mmol) in dichloromethane (50 mL) under argon, 1.27 mL (59.05 mmol) TEA, followed by the addition of 0.682 mL (4.5 mmol) of diethyl cyphosphonate (DECP ). Flash chromatography on silica gel gives 2.01 g of the desired compound. Example 3 N- (Cbz) amidino-2-aminovalerolactate.HCl-enasal- ((Z) -cArg.HCl): To a solution of 1.35 g (3.45 mmol) N- (Cbz ) amidino-2- (Boc) amino valerolactate (Example 1) in 25 ml of dioxane 15 ml of 3.38 M HCl in dioxane. The solution is stirred at ambient temperature for 1.5 hours, the solvent being removed. The residue is triturated with ether and the solids filtered under argon to give 1.05 g of white crystalline product. To a solution of 0.520 g (1.04 mmol) Boc-D-Phe-Phe-OBn (Example 2) in 10 ml of MeOH, 0.250 g of carbon and 0.5 ml of acetic acid. The mixture is hydrogenated in the PaaiM apparatus at 45-50 psi overnight. The catalyst is filtered through Celite (?) And the solvent is removed to provide 0.204 g of pre-duct. 23 35

65432 Case 4514

Example 5

Boc-D-Phe-Phe- (Z) -Arg: is added to a solution of Boc-D-Phe-Phe-OH (Example 2) (0.390 g, 0.980 mmol) and (Z) -Arg.HCl (0.932 g, 0.980 mmol) in dichloromethane to 0.30 mL (2.16 mmol) of TEA, below 0.17 mL (1.08 mmol) of diethyl dianophosphonate. The mixture is stirred overnight and washed with 1 N HCl, saturated sodium bicarbonate, and saturated sodium chloride. Chromatography on chill silica gives 400 g of product. = 0.45 (5% ipa / CH2 Cl2).

Example 6

Boc-D-Phe-Phe-Phe-Arg-H.OAc: is added to a cold solution of Boc-D-Phe-Phe- (Z) -Arg- (Example 5) (0.170 g, 250 mmol) in 15 mL of THF was added 0.019 g (0.500 mmol) of a LAH dose over the period of 1 minute. After stirring for 1 hour at this temperature. The reaction is quenched with 1 ml of water, warmed to ambient temperature and filtered, and the solvent removed. The residue is dissolved in dichloromethane and washed with brine. The resulting organic phase is dried over magnesium sulfate and filtered, the solvent removed. The resulting alcohol is dissolved in 10 ml of methanol, and 0.100 g of palladium on carbon is further added. The hydrogen is bubbled through the slurry for 1 hour, the mixture filtered through Celite®; the solvent is removed. The residue is dissolved in 7% water / acetic acid and lyophilized to provide 0.85 g of product. FAB-MS 533 / z (MH +), 575 m / z (MNa) +, 535 m / z (MH-E2 O) " 1 ".

Example 7

Boc-d-Pro-PH (Z) -Arg: is added to a solution of Boc-D-Pro-Phe-OH (0.357 g, 0.810 mmol) and (Z) -Arg-HCL in 24156532 Case 4514

25 ml of dichloromethane, 0.250 ml (1.78 mmol) of TEA dropwise, followed by 0.140 ml (0.890 mmol). The solution is stirred overnight, the solvent being removed. Chromatography on silica gives 0.362 g of product. R f = 0.40 (5% w / v / CH 2 Cl 2).

Example 8

Mod. 71-20,000 e *. - 90/08

Boc-D-Pro-Phe-Arg-H-OAc: as in Example 6, 0.285g 10 (0.460 mmol) of Boc-d-Pro-Phe- (Z) -Arg (example 7) and 0.035 g mmol) of LAH gives crude product. This product is chromatographed on silica gel to give 0.100 g of product, which is then hydrogenated (0.100 g of palladium on carbon) to provide 0.088 g of the desired product. Example 9 20

Boc-D-Nal-Oh (0.560 mmol) and 0.265 g of Phe-Obn.P-TSA (0.620 mmol) were treated with 0.100 ml of DECP 0.620 mmol) and 0.160 mL (1.12 mmol) gives 0.230 g of product. Rf = 0.75 (3% ipa / Ch 2 Cl 2).

Example 10 Boc-Dl-Nal-Phe- (Z) -Arg: as in Example 5, -0.æ0 30: g (0.270 mmol) of Boc-D1-Nal-Phe-OH (see Example 4 for the preparation of an analogous compound) and 0.122 g (Z) -Arg.HCl (0.270 mmol) with 0.076 ml of TEA (0.540 mmol) and 0.051 ml of DECP (0.300 mmol) gives 0.135 g of product. Rf = 0.50 (5% ipa / 30 / CH 2 Cl 2).

Example 11

Boc-Dl-Nal-Phe- (Z) -Arg (example 10) and 0.014 g of Boc-Dl-Nal-Phe- (Z) -Arg (Example 10) 4514 LAH (0.360 mmol) gives 0.078 g of chromatographed product, which is hydrophenated and lyophilized to give 0.049 g of product. FAB-MS 603 m / z (MH +). Example 12

Boc-D-Phe-Cha-OBn: as in example 2, 0.223 g (0.840 mmol) of Boc-D-Phe-OH and 0.250 g of Cha-OBn.HCl (example 76) (0.840 mmol) was treated with 0.140 ml (0.932 mmol) of DECP and 0.234 ml (1.68 mmol) of TEA gives 0.262 g of product. = 0.51 (25% EtOAc / pentane). Boc-D-Phe-Cha-OBn (example 12) and 0.110 g of palladium on carbon gave 0.212 g of product

Example 14

Boc-D-Phe-Cha- (Z) -cArg: as in Example 5, 0.165 g (0.507 mmol) (Z) -Arg.HCl (example 3) and 0.212 g (0.507 mmol) Boc-D-Phe (Example 13) with 0.085 mL (0.557 mmol) of DECP and 0.141 mL (1.01 mmol) of TEA gives 0.051 g of product. Rf = 0.62 (5% w / CH 2 Cl 3).

Example 15

Boc-D-Phe-Cha-Arg-H-OAc: as in Example 6, 0.049 g (0.071 mmol) of Boc-D-Phe-Cha- (Z) -Arg (example 14) and 0.005 g of LAH gives 0.020 g of chromatographed product which is hydrogenated (0.007 g Pd / C) and lyophilized to give 0.008 g of product. FAB-MS 559 m / z (MH +). 26 65432 dase 4514

10 15

Mod. 71-20,000 ex. - 90/08 25

Example 16

Doc-D-Phe-Phe-OBn: Boc-D-Phe-Phe-OBn (example 2) (0.500 g, 0.995 mmol) is dissolved in 5 ml of ethyl acetate, 5 ml of 5M HCl is added. After 2 , 5 hours, remove the solvent, the residue being placed in a vacuum pump overnight. The solid obtained (D-Phe-Phe-OBn.HCl) is dissolved in a mixture of 1 ml of dioxane and 0.995 ml of 1N NaOH. The solution is cooled to 0Â ° C and 0.092 g (1.09 mmol) and sodium bicarbonate are added. To this solution is added 0.092 g (1.094 mmol) of adamantyl fluoroformate in 1 ml of dioxane dropwise through a syringe. The solution was stirred at 0 ° C for half an hour and for 1.5 hours at room temperature. The solvent is evaporated, the residue being evaporated. is partitioned between ethyl acetate and water. The phases are separated, the aqueous phase being extracted with 3 x 20 ml of ethyl acetate. The combined organic phases are washed with brine, dried over magnesium sulfate and filtered, and the solvent is removed. The product is chromatographed on silica gel to give 0.390 g of the desired product. Rf = 0.70 (5% w / v Ch 2 Cl 2).

Example 17

Adoc-D-Phe-Phe-Phe-OH: As in Example 4, 0.380 g (0.653 mmol) of Adoc-D-Phe-Î'He-OBn (example 16) and 0.076 g Pd / C gave 0.192 g of the product required.

Example 18 30

Adoc-D-Phe-Phe- (Z) -cArg: as in Example 5, 0.100 g (0.2.04 mmol) of Adoc-D-Phe-Phe-OH (example 17) and 0.061 g (0.185 mmol ) of (Z) -cArg.HCl (example 3) with 0.031 ml (0.240) of DECP and 0.07 ml (0.408 mmol) of TEA gives 0.079 g of product, R f = 0.53 (5% ipa / CH 2 Cl 2 ). 27 15

Mod. 71-20,000 ex. - 90/08 20 25 30 65432 Case 4514

Example 19

Adoc-D-Phe-Phe-Arg-H.OAc: As in Example 6, 0.07æg (0.102 mool) of Adoc-B-Phe-Phe- (Z) -Arg (example 18) and 0.008 g (0.204 mmol) of LAH gives 0.064 g of product which is hydrogenated and lyophilized to give 0.059 g of product.FAB-MS 631 m / z (MH +). Example 20

2-Nal-Phe- (Z) -Arg: is added to 0.046 g (0.062 mmol) of Boc-D-2-Nal-Phe- (Z) -Arg (see example 10 for the preparation of an analogous compound) in 5 ml of ethyl acetate 1 ml of 5M HCl in ethyl acetate. The solution is stirred for 8 hours and the solvent is removed, the residue being placed in the vacuum pump for several hours. The resulting amine salt (D-2-Nal-Phe- (Z) -Arg.HCl) is dissolved in 0.5 ml of dioxane and 0.5 ml of water. 0.045 ml of 1N NaOH is added, followed by 0.004 g (0.049 mmol) of sodium bicarbonate. 0.010 g (0.049 mmol) of adamantyl fluoroformate in 1 ml of dioxane is added dropwise, the solution is stirred at 0 ° C for half an hour and at room temperature for 2 hours. The solvent evaporates and the residue is partitioned between ethyl acetate and water. The phases are separated, the aqueous phase being extracted with ethyl acetate. the organic phases are combined and washed with brine, dried over magnesium sulfate and filtered, the solvent removed. Flash chromatography gives 0.021 g of product. = 0.41 (5% w / CH 2 Cl 3). Example 21

Adoc-D-2-Nal-Phe-Arg-H.OAc: as in Example 6, 0.009 g (0.011 mmol) of Adoc-d-2-Nal-Pha- (z) -Arg (example 20) and 0.850 mg of LAH gives 0.007 g of product which is hydrogenated.

(0.002 g Pd / C) and leached to give 0.004 g of the desired product. FAB-MS 681 m / z (MH +).

Example 22

Boc-Phe- (Z) -Arg: Boc-Phe-OH solution (1.30 g, 4.90 mmol) and (Z) -Arg.HCl (example 3) (1.60 g, 4.90 mmol) in 25 ml of dichloromethane under an inert atmosphere, 1.43 ml (10.3 mmol) of TEA and 0.743 ml (4.90 mmol) of DECP are successively added. After stirring overnight the residue is chromatographed on silica gel to give 1.44 g of product.

Example 23

Phe- (Z) -cArg.HCl: Boc-Phe- (Z) -Arg (example 22) (1.44 g, 2.68 mmol) is dissolved in 20 ml of ethyl acetate and 22 ml of a solution 3M HCl in ethyl acetate is also added. The solution is stirred for 4 hours, the resulting solid material being filtered through a Buchner funnel with a continuous stream of argon passing over any exposed solid material. The filtered cake is washed with ethyl acetate and placed under vacuum for future use. 1.23 g of white solid are obtained.

Example 24

Boc-D-Trp- (Z) -Arg: is added to a solution of 0.16 g (0.320 mmol) Phe- (Z) -Arg.HCl (example 23 and 0.097 g (0.320 mmol) Boc- D-Trp-OH in 25 ml of dichloromethane in an inert atmosphere 0.090 ml of TEA (0.640 mmol), which was added 0.060 ml of DECP (0.360 mmol): the resulting solution was stirred overnight. The solvent is removed and the residue is chromatographed on silica gel to give 0.160 g of product, Rf = 0.40 (5% w / CI 3).

Mod. 71-20,000 ex. - 90J08 25 * 65432 Case 4514

Example 25

Boc-P-Trp-Phe-Arg-H.OAc: as in Example 6, 0.110 g (0.150 mmol) of Boc-P-Trp-Phe- (Z) -Arg (example 24) and 0.011 g of LAH gives 0.065 g of chromatographed product, which is hydrogenated (0.050 g Pd / C) and lyophilized to give 0.040 g of product. FAB-MS 592 m / z (MH +), 700 m / z (MH + TG) + 492 m / z (MH-Boc) +.

10.0 g (95.2 mmol) of D-Serine and 4.78 g (76.12 mmol) of sodium cyanoborohydride in 200 ml of methane , and 10.6 g (104 mmol) of benzaldehyde was added dropwise. The mixture is stirred for 48 h and filtered. The filter cake is washed with methanol and placed in a vacuum pump overnight. 10.5 g of white powder is obtained which is a 1: 1 mixture of the starting material and the product.

A 1: 1 mixture of n-Bn-P-Ser-OH and P-Serine (example 26) (5.00 g) was dissolved in 18.3 ml of 2N NaOH, further adding 6 ml of THF. The solution is cooled to 0 ° C, where 7.86 mL (55 mmol) of benzylchloroformate is added. during the addition the pH is maintained between 9.5 and 10.5 by addition of 1N NaOH. After 1 hour of stirring and continuous pH maintenance, the solution is diluted to pH = 2.4 with concentrated HCl. The acidic solution is extracted with ethyl acetate (3X) and dried with magnesium sulfate; the solvent being removed. The residue is chromatographed. dried over silica gel to give 4.00 g of the desired product. = 0.27 (1% acetic acid / 5% ipa / 94% dichloromethane). 30 35 65432 Case 4514

Example 28 N-Bn-N- (Z) -D-Ser-B-lactone: is added to a dry flask under argon containing 1.52 g of triphenylphosphine (5.77 mmol) in 20 ml of THF at 78 ° C, 0.637 ml of dimethylazodicarboxylate (5.79 mmol) dropwise. After 15 minutes, a solution of N-Bn-N- (Z) -D-Ser is added and the solution is allowed to warm to room temperature overnight. The solution is evaporated, the residue chromatographed on silica gel to give 1.00 g of product. Rf = 0.37 (208 of ethyl acetate / petroleum ether).

To a mixture of dimethylsulfoxide complex cuprous bromate (0.581 g, 2.83 mmol) in 5 mL of THF was added to a pre-dried flask aq. 78.9 under argon, 2.96 ml of t-BuLi (1.7M in pentane, 5.04 mmol) dropwise. The mixture is stirred at this temperature for 45 minutes and for 20 minutes at 45 ° C. The mixture is recooled to -78 ° C, and N-Bn-N- (Z) -D-Ser-B-lactone (Example 28) (0.200 g, Q 642 mmol) in 2 mL of THF is added. After stirring for 16 hours at -78Â ° C, the mixture is poured into cold 1N HCl, methanol (25 volume%) being added. After 20 minutes the mixture is filtered, the filtrate is extracted with ethyl acetate and dried with magnesium sulfate; the solvent is removed. Chromatography of the residue on silica gel gives 0.120 g of product. + 32.18 (c = 1, chloroform).

R f = (18 HCOA / 5% MeOH / 94% CH 2 Cl 2).

Example 30

Boc-D-t-Bal-OH: is added to a solution of N-Bn- (Z) -D-tBal-OH (example 29) (0.160 g, 0.432 mmol) in water:

Mod. 71-20,000 ex. - 90 (08 20 25 30 65432 Case 4514

(1: 2, 15 ml), 0.030 g Pd / C and the hydrogenated paste in the Paarâ "¢ machine at 45 psi for 16 hours. After filtration through Celite®, the solvent is removed, the residue being placed " in the vacuum pump. The deprotected product is dissolved in 0.480 ml of 1N NaOH and 2 ml of dioxane, 1 ml of water is added. The solution is cooled to 0Â ° C and 0.480 ml of 1N NaOH and 0.078 g di-t-butyldicarbonate (0.465 mmol) in 1 ml of dichloromethane are simultaneously added in separate addition funnels, the resulting solution being stirred for 1 h at 0Â ° C. The solution is extracted with ethyl acetate, the aqueous phase being reacted to pH = 2.5. Extraction of the aqueous phase with ethyl acetate is followed by drying with magnesium sulfate, drying and removal of solvent. The residue is chromatographed over silica gel to give 0.109 g of pure material [α] D = + 32.7 (c = 1, chloroform). Example 31 Boc-Dt-Bal-Phe-cArg: As in Example 24, 0.040 g of Boc-Dt-Bal-OH (0.163 mmol) and 0.077 g (0.163 mmol) of Phe- (Z) -Arg.HCl (example 23) with 0.045 mL (0.326 mmol) of TEA and 0.025 mL (0.163 mmol) of DECP gives 0.055 g of product. Example 32 Boc-Dt-Bal-Phe-arg-OAc: as in Example 6, 0.055 g (0.083 mmol) of Boc-Dt-Bal-Phe (10% MeOH / 90% CH2 Cl2) - (Z) -Arg (example 31) and 0.006 g (0.166 mmol) of LAH gives 0.027 g of product which is hydrogenated (0.010 g Pd / C) and lyophilized to give 0.035 g of product. FAB-MS-533 m / z (MH + 6.555 m / z (MNa) +; 641 m / z (MH + TG) + 515 m / z (MH-H2 O) + 32 35 65432 Case 4514

15

Mod. 71-20,000 e *. - 90/06 20

Example 33 N-Bn-N- (Z) -Dt-Ad-OH: as in Example 29, 5.20 mL (4.69 mmol) 1-adamantyl bromide magnesium, 0.454 g (2.41 mmol) cuprous / dimethylsulphide complex and 0.031 g (0.963 mmol) N-Bn-N- (Z) -Ser-lactone (example 28) in 0.514 ml of methyl sulfoxide and 15 ml of THF gives 0.333 g of product. = + 20.3 (c = 1, chloroform). = 0.25 (1% HCO / 5% ipa / 94% CH 2 Cl 2).

Example 34

Boc-D-Adl-OH: as in Example 30, 0.160 g of N-Bn -N- (Z) -D-1-Agl-OH (example 33) (0.358 mmol) and 0.032 g Pd / C gives 0.081 g of product as white powder, 0.60 g of this product is protected with Boc compound with 0.115 g of di-t-butyl dicarbonate (0.296 mmol) and 5.37 ml IN NaOH to give 0.047 g of product . DG D = + 3.19 (c = 1 · chloroform). Rf = 0.38 (1% HOAc / ipa / 94% CH 2 Cl 2).

Example 35

Boc-Dl-Adl-Phe- (Z) -cArg: as in Example 24, 0.047 g (0.145 mmol) of Boc-D-1-Adl-OH (example 23) with 0.069 g (0.145 mmol) of Phe- (Z) c Arg HCl (example 23) with 0.044 ml (0.319 mmol) TEA and 0.022 ml (0.145 mmol) DECP gives 0.067 25 g of product. D = -15.8 (c = 1, chloroform). Rf = 0.64 (5% ipa / CH 2 Cl 2).

Example 36

Boc-Dl-Adl-Phe- (Z) -Arg (example 35) (0.088 mmol) and 0.007 g ( (0.002 g Pd / C), which is lyophilized to give 0.015 g of product FAB-611 m / z (MH +), 719 m / z ( MH + 33 30 to 15

Mod. 71-20,000 ex. - 90/08 20 25 30 35 6 5432 Case 4514

/ TG) +.

Example 37 [- NÎ ± - (Z) -D-Orn-Phe- (Z) -Arg: as in Example 24, 0.250 g (0.710 mmol) of N-Boc-N- ( Z) -D-Orn-OH and 0.338 g (0.71 mmol) of Phe- (Z) -Arg.HCl (example 23) was treated with 0.330 mL (2.80 mmol) of TEA and 0.120 mL (0.800 mmol) of DECP gives 0.310 g of product. = 0.52 (5% ipa / Cl 2 Cl 3): Example 38 N-Adoc- <N- (Z) -D-Orn-Phe- (Z) -Arg; To a solution of 0.300 g (0.420 mmol) of (N-Adoc-N- (Z) -D-Orn-Phe- (Z) -Arg, (Example 37) in 15 ml of water and 15 ml of dioxane at a temperature of 0 ° C is added 0.134 g (1.60 mmol) of sodium bicarbonate, followed by 0.099 g (0.500 mmol) of adamantyl fluorformate in 1 ml of dioxane dropwise. The organic phase is washed with brine, dried over magnesium sulfate, and the solvent removed. Chromatography (MgSO 4) is then added and the solvent is removed under reduced pressure. on silica gel affords 0.120 g of product &lt; Rf = 0.52 (5% w / CH 3 Cl 2).

Adoc-D-Orn-Phe-Arg-H-OAc: as in Example 6, 0.165 g (0.190 mmol) of N -Doc-N- (Z) -D-Orn-Phe- (Z ) -c Arg (Example 38) and 0.015 g (0.380 mmol) LAH gives 0.035 g of product which is hydrogenated and lyophilized to give 0.030 g of product.

Example 40

Adac-D-t-Bug-Phe-OBn. is added to a 5 ml solution 34 6.5432 Case 4514

10

Mod. 71-20,000 ex. - 90/08 20 25

Example 40

Adac-D-t-Bug-Phe-OBn: To a solution of 5 ml of a phosgene solution in toluene under argon is added 0.140 ml of TEA (1.00 mmol). To this is followed the addition of 0.112 g (0.750 mmol) of 1-adamantanamine in 1 ml of toluene dropwise through a syringe. The resulting mixture is stirred for 20 minutes, with excess phosgene removed through the action of argon. The solid is drawn by suction through a sintered glass funnel, the solvent being removed. The residue is redissolved in dichloromethane and cooled to 0 ° C. To this solution is added 0.140 ml of TEA (1.0 mmol), followed by 0.100 g (0.250 mmol) of D-t-bug-Phe-OBn.HCL (see example 16 for preparation of an analogous compound) in one portion. The resulting solution is stirred for 2 hours at room temperature, the solvent removed. Chromatography on silica gel gives 350 g of product.

Example 41

Adac-Dt-Bug-Phe- (Z) -g Arg: is added to a mixture of Adac-Dt-Bug-Phe-OH (see Example 4 for preparation of an analogous compound) (0.150 mmol) and Z) -Arg.HCl (example 3) (0.080 g, 0.180 mmol) in 25 mL dichloromethane under argon was added 0.084 mL (0.600 mmol) TEA, hereinafter 0.030 mL (0.200 mL) DECP. The mixture is stirred one day to the next, the solvent being removed. The residue was chromatographed on silica gel to give 0.085 g of product.

Rf = 0.35 (3% ipa / CH2 Cl2).

Example 42

Adac-Dt-Bug-Phe-Arg-H: as in Example 6, 0.085 g Adac-Dt-Bug-Phe- (Z) -Arg-H (example 41) (0.120 mmol) and 0.009 g LAH 0.240 mmol) gives 0.054 g of product chromate-35

65432 Case 4514 which is hydrogenated and lyophilized to give 0.044 g of product. FAB-MS = 596 m / z (MH +).

Example 43 10 15

Mod. 71-20,000 eic. - 90/08 20

Ada-Dt-Bug-Phe- (Z) -Arg: is added to a solution of Boc-D-Bug-Phe- (Z) -g Arg (see example 5 for method of preparation of an analogous compound) (0.060 g , 0.082 mmol) in 2 ml of ethyl acetate was added 1 ml of 5 M HCl in ethyl acetate, the solution stirred at room temperature for 6 hours. Further, 1 ml of HCl in ethyl acetate is added, the solution being stirred overnight. The solvent is removed, the residue being placed in a vacuum pump. The solid material is dissolved in 20 ml of dichloromethane, 0.018 g (0.091 mmol) of adamantaneecacetyl being added as a chloride. To this solution is added 0.025 ml of TEA (0.180 mmol) followed by 0.014 ml of DECP (0.091 mmol), whereupon the resulting mixture is stirred overnight. Removal of the solvent and chromatography on silica gel give 0.013 g of compound = 0.52 (5% MeOH / CH 2 Cl 2).

Example 44

Ada-Dt-Bug-Phe-Arg-H: as in Example 6, 0.013 g 25 (0.018 mmol of Ada-Dt-Bug-Phe-cGARG (example 43) and LAH (0.0015 g, 0.036 mmol) gives 0.002 g, which is hydrogenated and lyophilized to provide 0.002 g of product FAB-MS - 595 m / z (MH +), 577 m / z (MH-H 2 O) + Example 45 (4R) -3- (2) adamantyl oxo-4- (phenylmethyl) -2-oxazolidinone: To a solution of 1.00 g (6.06 mmol) of (4 R) - (phenylmethyl) -2-oxazolidinone in 30 ml of THF at -78 DEG C. 2.70 ml of lithium butyl (6.67 mmol) was added dropwise after the addition of 365 65432.

The solution is stirred for 10 minutes, followed by the dropwise addition of 1.53 g of acetyl chloride adamantane (7.25 mmol) in 5 ml of THF. After 15 minutes the cooling bath is removed, the solution is allowed to warm to room temperature, the reaction is cooled with 2 ml of water and concentrated. The residue is partitioned between ether and water, the phases separated. The organic phase is dried and the solvent is removed. Chromatography with 10% ethyl acetate / pentyl ether on silica gel gives 1.62 g of desired product as a white solid.

Example 46 (3- (2-Amino-2-oxazolidinone) -3- (2-adamantyl-2-azido-1-oxo) -4- 1-oxo) -4- (phenylmethyl) -2-oxazolidinone (example 45) (0.0389 g, 1.10 mmol) in 15 mL of THF is cooled to -78 DEG C. This solution is then added via channels to a 0.3 M THF solution of 1.27 mmol KHMDS, which is also lied at -78 DEG C. After stirring for 0.5 h at this temperature, a 78 C solution of trisyl azido (cf. Harrison, RE, G. Cellmen & SKGrupta, &quot; The Reaction of Arylsulfonyl Azides with N-Methylindole &quot;, Journal of Organic Chemistry, Vol. 38, No. 111 (1973), pp. 11-16). The solution is diluted with dichloromethane (5 ml) and the solution is partitioned between ethyl acetate and dichloromethane dichloromethane and washed with brine and saturated sodium bicarbonate after drying (magnesium sulfate) and filtering The residue is chromatographed on silica gel to provide 0.340 g (0.860 mmol) of the desired product as a colorless oil.

Example 47 (2R) -2-azido-adamantaneacetic acid; 37 6.5432 Case 4514

1- (3 (2R), 4R) -3- (2-adamantyl-2-azido-1-oxo) -4- (phenylmethyl) -2-oxazolidinone (example 46) (0.270 g, 0.690 mmol) This material is dissolved in 10 ml of THF, which is cooled to 0 ° C. To this solution is added a mixture of 3.63 ml of 0.2 M lithium hydroxide, 35 ml of 30% hydrogen peroxide and 4 ml of water, dropwise. After 1 hour at 0 ° C, 10 equivalents of sodium bisulfite are added in 2 ml of water, the solution is stirred for 15 minutes. The THF is removed, the remaining aqueous solution being extracted 3 times with ethyl acetate and the organic phase is dried over magnesium sulfate. After fixation, the solvent is removed, the residue being chromatographed on silica gel to provide 0.135 g of the product as a white solid. 15

Mod. 71-20,000 ex. - 90/08 25

Example 48 (R) -Adoc-Ad-OH: 2-azido-adamantane: ethical acid (example 47) (0.070 g, 0.340 mmol) is dissolved in 2: 1 acetic acid / water (2 ml), adding 0.150 g of? D / C. The mixture is hydrogenated in the apparatus Paarx at 50 psi for 16 h. the mixture is filtered through Celite®, the solvent is removed. The residue is dissolved in 0.34 ml of 1N NaOH and 2 ml of water, and 2 ml of dioxane is added. To this solution is added 0.027 g of sodium bicarbonate, to which is added an addition of fluoroformate adamantyl (0.101 g, 0.510 mmol) in 1 ml of dioxane. After 30 minutes, the cooling bath is removed, the solution being warmed to room temperature. The mixture is diluted with acetate, the phases being separated. The aqueous phase is acidified to pH = 2 and extracted with ethyl acetate. Removal of solvent gives 0.040 g of product.

Example 49

Adoc-D-Ad-G-Phe- (Z) -g Arg: as in Example 24.0.040 g (0.100 mmol) of (R) -Adoc-Ad-G-OH (example 48) and 0.049 g

(Example 23) with 0.056 ml (0.400 mmol) of TEA and 0.018 ml (0.120 mmol) of DECP gives 0.031 g of product after chromatography. Phe- (Z) -Arg.HCl (example 23) = 0.65 (5% ipa / CI 3 Cl 2) 5

Example 50

Adoc-D-Ad-Phe- (Z) -g Arg (Example 49) (0.037 mmol) and LAH (0.003 g, 0.075 mmol ) affords 0.10 g of chromatographed product which is hydrogenated with 0.002 g Pd / C and lyophilized to provide 0.009 g of product. FAB-MS 675 m / z (MH +).

Example 51

Mod. 71-20,000 ex.

Adoc-Dt-Bug-Phe- (Z) -Lys-OMe: is added to a solution of 0.432 g of Adoc-Dt-Bug-Phe-Phe-OH Adoc (see example 17 for preparation of an analogous compound) (0.946 mmol) and 0.298 g NE- (Z) -lys-OMe (0.901 mmol) in 25 mL of cilomoromethane was added 0.276 mL of TEA (1.98 mmol), to which 0.144 mL of DECP (0.946 mmol), the solution being stirred overnight. The solvent was removed, the residue chromatographed on silica gel to give 0.495 g of product. Rf = 0.49 (4% MeOH / CH 2 Cl 2). 25

Example 52

Adoc-Dt-Phe- (Z) -Lysinol: Adoc-Dt-Bug-Phe- (Z) -Lys -OMe (example 51) (0.070 g, 0.096 mmol) is dissolved in 1 ml of ethanol and cooled in an ice / methanol bath. To this solution is added anhydrous calcium chloride (0.021 g, 0.191 mmol) and 1 mL of THF. Sodiumborohydride (0.144 g, 0.382 mmol) is added in one portion, leaving the resulting slurry The reaction is quenched with 1 ml of 1M citric acid, the solvent is removed, the residue is partitioned between ethyl acetate and water.

Mod. 7! - 20,000 e *. - 90/08 20 25 30 (&gt; 5432 Case 4514

is washed with brine and dried, the solvent removed. Silica gel chromatography gives 0.042 g of the desired compound [Î'] D = + 14.8 (c = 1, EtOH).

Example 53 Adoc-D-t-bug-Phe- (Z) -Lys -h. is added to a solution of 0.026 ml of oxalyl chloride (0.275 mmol) in 5 ml of dichloromethane at -78 DEG C., 0.042 ml of DMSO (0.570 mmol / in 0.500 ml of dichloromethane dropwise). After 5 minutes, 0.160 g of Adoc-Dt-bug-Phe- (Z) -Lys-Me (Example 51) (0.227 mmol) in 1 ml of dichloromethane is added.The solution is stirred at -78 ° C for 20 minutes, ml of TEA The solution is then warmed to 0 ° C and cooled with 2 ml of water The residue is washed with 1 N HCl, saturated sodium bicarbonate and brine, the organic phase is dried and filtered, the solvent removed Chromatography on silica gel of the residue gave 0.096 g of desired product. [Α] D = -45.3 (c = 1, CHCl 3) Example 54 Adoc-Dt-Bug-Phe-Lys- Tfa: is added to a solution of 0.009 g of Adoc-t-Bug-Phe- (Z) -Lys-H (example 53) (0.013 mmol) 3 0.006 g of 9.7% HCl in dioxane in 0.900 ml of DMF 0.005 g of black Pd This paste is hydroganged at atmospheric pressure for 20 minutes. This is diluted in 1 ml of dioxane and filtered through Carrier. The filter cake is washed with methanol, the solvent removed. The residue is reverse phase chromate-rhaphdn (water / acetonitrole / 0.1% TFA) to give 0.008 g of the desired product. FAB-MS - 569 m / z (MH +), 677 m / z (MH + TG) +. Example 55 eBroc-D-t-Bug-Phe- (Z) -Arg: is added to a solution of 40

Mod. 71-20,000 ex. - 90/08 20 25 30 65432 Case 4514

0.080 g of (-) endobroneol (0.520 mmol) and 0.41 ml of pyridine (0.520 mmol) in 2 ml of toluene, a solution of 12% phosphocholine in toluene dropwise. After 1 hour, the argon is bubbled through a solution to clear any remaining phosgene. After 10 minutes, the mixture is filtered, the solvent being removed. The residue is dissolved in dichloromethane, adding 0.127 g of Dt-Bug-Phe- (Z) -Arg.HCl (see example 20 for the preparation of an analogous compound) (0.240 mmol), which is then 0.100 ml (0.720 mmol) of TEA. The solution is stirred overnight, with solvent removed. Chromatography on silica gel gives 0.100 g of the desired product. (5% w / CH 2 Cl 3) = 0.75. EBroc-Dt-Bug-Phe-Arg-H: as in Example 6, 0.090 g (0.120 mmol) of eBroc-Dt-Bug-Phe- (Z) -Arg (example 55) and 0.009 g of LAH ( 0.240 mmol) gives 0.065 g of product after chromatography. This compound is hydrogenated in the apparatus for 3 hours (AcOH / MeOH-1/20) to give 0.045 g after filtration. FAB-MS -599 m / z (MH +), 707 m / z (MH + TG) +. 26.0 g (99.8 mmol) of p-N ph -OMe-HCL in 500 ml of dichloromethane are suspended and 27.8 ml of TEA (199.6 g) are added. mmol) dropwise. After the addition is complete, 26.4 g (94.8 mmol) of trityl chloride in 250 ml of dichloromethane is added. The yellow solution is stirred for 2 hours with brine, dried over magnesium sulfate and filtered, and the solvent is removed to give 44.0 g of product, yielding a purity suitable for use in the reaction of Example 58. 41 35 $ 5432 Case 4514

5 10 15

Mod. 71-20,000 ex. - 90/08 20 25

Example 58

Tr-p-nitrophenylethanol: is added to a solution of 45.3 g (97.2 mmol) of Tr-p-Nph-OMe (example 57) in 500 ml at 0 ° C under argon 200 ml (300mmol) of DIBAL-H at a rate moderate After 1 hour, the solution is transferred to a 2 liter Erlenmeyer flask cooled to 0Â ° C. To this solution is added 500 ml of a saturated potassium sodium tartrate solution (very cautiously). The mixture is extracted with 3 X 11 of ethyl acetate, the organic phases being dried. Removal of the solvent gives 40 g, which is chromatographed on silica gel to give 26.9 g of the product.

To a solution of 0.581 g of oxalyl chloride (5.57 mmol) in 50 ml of dichloromethane in argon at -78 ° C was added 0.946 ml of DMSO (13.3 mmol) in 2 ml dichloromethane, dropwise. After the addition is complete, Trp-p-nitrophenylalaninol (example 58) (2.25 g, 5.13 mmol) in dichloromethane (7 mL) is added. The solution is stirred for half an hour, then poured into a mixture of ether and water. The phases are separated, the organic phase is washed with water, dried with magnesium sulfate and filtered; the solvent is removed. The residue is chromatographed on silica gel to provide 2.00 g of product. = 0.55 (28 CH 2 C (O) CH 2 /

/ CH 2 Cl 2), [t D 25 = + 67.8 (c = 1, CH 2 Cl 2)

To a solution of 2.23 g of Trp-p-nitrophenylalaninol (example 59) in dichloromethane (2 ml) was added triethylamine (5.11 mmol) in 100 mL of THF under argon was added 0.951 g of CFâ,ƒMSO-D-dimer (6.13 mmol), followed by addition of 0.161 g of tetrabutylammonium fluoride trihydrate

65432 Case 4514 (0.510 mmol); the solution is stirred for 2 H. The solvent is removed, the residue chromatographed on silica gel to provide 1.73 g of the product. Rf = 0.50 (2% CH₂C (O) CH / / CH₂Cl₂).

Example 61 10

Mod. 71-20,000 ex. 3-Amino-4-p-aminophenyl-1,1-trifluoro-2-butane, is added to a solution of 4-p-Nitrophenyl-1,1,1-trifluoro-3-tritylamino -2-butanol (example 60) (1.00 g, 1.98 mmol) in 50 mL of ethanol, 2.23 (9.88 mmol) of sodium chloride dihydrate. The mixture is poured for 1 hour and poured into ice water (100 ml). The pH is adjusted to 8.4 with sodium bicarbonate, and the mixture is extracted with ethyl acetate (2 x 250 ml). The organic phase is dried and filtered, the solvent being removed. Silica gel chromatography gives 0.800 g of product. Rf = 0.18, 0.30 (1% NH 4 OH / 10% MeOH / 89% CH 2 Cl 2).

Example 62 4-p-Aminophenyl-3-t-butyloxycarbonylamino-1,1-trifluoro-2-butane: is added to a solution of 3-amino-4-p-aminophenyl-1,1,1- (5.26 g, 22.5 mmol) in 200 mL of THF (5.31 g) of di-t-butyl dicarbonate (23.6 mmol) was added the mixed solution The solvent was removed, the residue chromatographed on silica gel to give 5.70 g of product. Rf = 0.48, 0.68 (1% NH4 OH / 1% ipa / 98% CH 2 Cl 2).

Example 63 3-t-Butyloxycarbonylamino-4-di-CBz-p-guanidonophenyl-1,1,1-trifluoro-2-butane; is added to a solution of 4-p-aminophenyl-3-t-butyloxycarbonylamino-1,1,1-trifluoro-2-butanol (Example 62) (0.020 g, 0.060 mmol) in THF (5 mL) under argon. 0.027 mg (0.072 mmol) of N, N ', di-Cbz-S-methyl-isothiourea (example 74), followed by the addition of 5.7 mg of mercuric acetate. The mixture is stirred overnight with the addition of 5.7% mercuric acetate the next morning. After 1 hour the THF is evaporated, the residue redissolved in ethyl acetate. The organic phase is washed and saturated with sodium bicarbonate, 0.1N HCl and brine. The solution is dried and filtered, the solvent being dissolved. Chromatography on silica gel gives 0.035 g of product. Rf = 0.28 (3% ipal / CH 2 Cl 3).

Example 64

3-Amino-4-di-Cbz-guanidinophenyl-1,1,1-trifluoro-2-butanol hydrochloride salt: A solution of 2.8 g (4.34 mmol) of 3 t -butyloxycarbonylamino- 1,1-trifluoro-2-butanol (example 63) in 40 ml of dioxane and 15 ml of 4 M HCl in dioxane was heated at reflux for 2 hours at the temperature environment. The dioxane is removed in vacuo, the residue triturated with ether. The slurry is filtered under argon and weighs 2.4 g.

Example 65 3- (Adamantiyloxy-D-t-butylglycylphenylalanyl) amino-4-di-Cbz-p-guanidinophenyl-1,1,1-trifluoro-2-butanol: is added to a solution of 0.750 g of 3-amino- 1,1,1-trifluoro-2-butanol (example 64) (1.29 mmol) and 0.647 g of Adoc-Dt-Bug-Phe-OH (see example 17 for the preparation of an analogous compound) (1.42 mmol) in 10 mL of DMF is added 0.500 mL of TEA (3.58 mmol), which is followed by 0.232 mL of DECP, (1.42 mmol); the mixture is stirred overnight. The solvent is removed and the chromatographed residue (40% ethyl acetate / ether petroleum) on silica gel to yield 0.890 g of product. Rf = 0.70 (5% 44 65432 Case 4514

1a / CH2Cl2).

Example 66 5 10

Mod. 71-20,000 ex. (Adamantiloxy-D-butylglycylphenylalanyl) amino-4-di-Cbz-p-guanidinophenyl-1,1,1-trifluoro-2-butanone: is added to a solution of 0.452 g of (1.07 mmol) in 10 ml of dichloromethane, 0.352 g of 3- (adamantyloxy-D-butylglycylphenylalanyl) amino-4-di-Cbz-p-guanidinophenyl-1,1,1 -trifluoro-2-butanol (example 65) (0.356) in 10 ml of dichloromethane was added dropwise. The red-brown solution is stirred for 2 hours and is diluted with 30 ml of ether. The mixture is poured into 50 ml of saturated sodium bicarbonate containing 1.85 g of sodium thiosulfate. This solution is stirred for 10 minutes, adding 30 ml of ether. the phases are separated, the organic phase is dried over magnesium sulfate. The solvent is removed, the residue chromatographed on silica gel to give 0.120 g of product. = 0.52 (75% EtOAc / pentane), â € ƒâ € ƒâ € ƒ32.3 (c = 1, CH2 Cl2). Example 67 3- (Adamantiloxy-D-butylglycylphenylalanyl) amino-4-p-guanidinophenyl-1,1,1-trifluoro-2-butanone: is added to a solution of 0.120 g of 3- (adamantyloxy-D-butylglycylphenyl- 1,1-trifluoro-2-butanone (Example 66) (0.122 mmol) in 20 mL of methanol and 0.5 mL of acetic acid was added 0.025 g Pd / C, the slurry being hydrogenated at 45psi for 1.5 hours the slurry is filtered through Celite, the solvent being removed. The reverse phase HPLC (acetonitrile / water / TFA) gives 0.017 g of a diastereomer and 0.028 g of the other. FAB-MS-732 m / z (MH + + H 2 O), 746 m / z (MH + + CH 3 OH). 45 35 v $ 5 10 15

Mod. 71-20,000 ex. -90/08 65432 Case 4514

Example 68 4-t-Butyloxycarbonylamino-nitributane: is added to a solution of 4.00 g of 4-t-butyloxyamino-1-aminobutane (see Stahl GL, R. Walter and CW Smith, &quot; General Procedure for Synthesis of 1 , Mono-N-acylated 6-Diaminohexanes, quot; Journal of Organic Chemistry, Vol. 43, No. 11 (1978) pp. 2285-2286) (21.3 mmol) in 50 ml of dichlorethane was added 14.7 g (85.0 mmol ) PCPBA in 1 g portions. The temperature is raised to reflux (83 ° C) and held there for 3 h. A further 7.80 g (45.0 mmol) of MCPBA is added and reflux continued for more than one hour. The solution is quenched and washed with 3 x 30 mL of 1N NaOH. The organic layer is dried (magnesium sulfate), the solvent removed. Chromatography on silica gel gives 1.90 g of product.

Example 69 6-Boc-amino-3-nitro-1,1,1-trifluoro-2-hexanol: is added to a solution of 0.500 g (2.29 mmol) of 4-t-Boc-amino-1 (Example 68) and 0.239 g (2.08 mmol) of trifluoroacetic acid hydrate in 10 mL of THF, 0.013 g of N, N-dimethylformamide were added. kneading potassium potassium. The slurry is heated to 50Â ° C and stirred at this temperature overnight. The slurry is dissolved in ether and washed with brine. The organic phase is dried, the solvate being removed. Chromatography of residue gives 0.38 of product as a mixture of diastereomers. Rf = 0.34 (5% w / C 1 H 3)

Example 70

3-Amino-6-Boc-amino-1,1,1-trifluoro-2-hexanol: is added to a solution of 0.280 g (0.885 mmol) of 6-Boc-amino-3-nitro- , 1-trifluoro-2-hexanol (example 69) in 5 ml of methanol, 5 drops of acetic acid and 0.056 g of 10% Pd / C. A 46 35

65432

The paste is hydrogenated at 45 psi for 20 h and filtered through Celitev. The solvent is removed, the residue is chromatographed on silica gel to give 2 diastereomers (0.072 g and 0.108 g) which are readily separable. Rf = 0.27, 0.45 (1% MH + OH / 2.5% MeOH / 96.5% CH 2 Cl 2).

Example 71 3- (Cyclohexyl) -D-Bug-Phe-amino-6-t-butyloxycarbonylamino-1,1,1-trifluoro-2-hexanol: is added to a solution of 3-amino-6- 1,1-trifluoro-2-hexanol (example 70) (0.60 g, 1.31 mmol) and eBroc-Dt-Bug-Phe-OH (see example 17 for the preparation of an analogous compound) (0.370 g, 1.31 mmol) in dichloromethane (20 mL) under argon was added THF (2.62 mmol), followed by addition of 0.200 mL of DECP (1.31 mmol). The residue is chromatographed on silica gel to provide 0.800 g of product: 0.34 (5% w / CH 2 Cl 2).

Example 72 3- (eBroc-β-Bug-Phe) -amino-6-guanidino-1,1,1-trifluoro-2-hexanol of TFA salt: is added to a solution of 0.800g 3- bromo-D-Bug-Phe) amino-6-Boc-amino-1,1,1-trifluoro -hexanol (example 71) (1.10 mmol) in dioxane (20 ml), and 3.2M HCl in dioxane; the mixture being stirred for 2 hours. Additional 1 ml of HCl is added at this point, the mixture stirred for an additional 2 hours and 30 min. The solvent is removed, the residue being placed in the vacuum pump overnight. The solid material is dissolved in 50 ml of dry DMF, the pH adjusted to 8.5 with TEA. 7.6 g (37.8 mmol) of 3,5-dimethylpyrazole-1-carboxamidine nitrate (DMPCNO) was dissolved in 5 ml of DMF in a vial, the pH adjusted to 8.5. the two solutions are combined and stirred for 5 days. The pH is adjusted to 5.8 with acetic acid 47 65432 Case 4514 5 10

Q being the solvent removed. The reverse phase of HPLX purification of the residue gives 0.190 g of the desired product.

Example 73

Mod. 71-20,000 ex. - 90/08 TFA salt 3- (eBroc-D-Bug-Phe) -amino-6-guanidino-1,1,1-trifluoro-2-hexanone is added to a solution of Period- Martin (0.400 g, 850 mmol) in 10 mL CH 2 Cl 2 a solution of 3- (eBroc-D-Phe) -amino-6-guanidino-1,1,1-trifluoro-2-hexanol of TFA salt (ex. ) (0.190 g, 0.240 mmol) in 10 mL of CH2 Cl2. The mixture is stirred overnight, the solvent being removed. The residue is redissolved in CH 2 Cl 2 and washed with saturated NaHCO 3 solution. The solvent is removed from an organic phase, the residue dissolved in 2 ml of MeOH and passed through a Biorad-AGB1X8 / 200-400 meshes-acetate column. After removal of the solvent, the residue is chromatographed (HPLC reverse phase) to provide 0.080 g of product. 20

Example 74 N, N'-bis-carbobenzyloxy-s-methyl-isothiourea: is added to a solution of S-methyl-isothiourea dimer sulphate (5.00 g, 18.0 mmol) (36.0 mL, 36.0 mmol), 11.3 mL (79.0 mmol) of benzyl chloroformate and 79.0 mL (79.0 mmol) of IN NaPH were added dropwise from separate addition funnels. The slurry is stirred for half an hour at 0 ° C and 1.5 hours at room temperature. The solution is extracted with dry ethyl acetate with magnesium sulfate and the filtrate is evaporated. Chromatography on silica gel gives 0.262 g of product. Rf = 0.40 (15% EtOAc / pet ether). Example 75 35

Boc-Cha.OBn: is added to a mass of Boc-Cha-OH (1.56 48

(1.54 g, 18.33 mmol) in 50 mL of DMF, 0.700 mL (6.72 mmol) of benzyl bromide. The mixture is stirred in EtOAc. The slurry is washed with water, the organic phase is dried and the filtrate is evaporated. Recover 1.91 g of product which is homogeneous by TLC. = 0.70 (25% EtOAc / pet ether).

Example 76

Cha-OBn.HCl: is added to a solution of Boc-Cha-OBn (1.81, 5.01 mmol) (example 75) in dioxane, 18 ml of 3.34 M HCl in dioxane. After 1 hour, an additional 10 ml of 3.34 M HCl in dioxane is added. The solvent is removed, the residue is triturated with EtOAc, filtered and placed in a vacuum pump. Recover 1.22 g of white solid.

MET0D0S FOR COMPONENT ACTIVITY TESTING

The following non-limiting procedures are methods for testing the anti-inflammatory and / or analgesic activity of the tripeptide derivatives of this invention. There are various inhibition experiments of enzymes which are known to be predictive of the anti-inflammatory activity of compounds. Such enzyme experiments are useful for measuring the activity of the compounds of this invention. Such enzyme experiments include the following: porcine pancreatic kallikrein (PKK) - see references of A., E. and F; Human urinary kalicretium (HUK) - see references E and F; kalicrein human plasma - see references B and E; human plasmin (HP) -see references B and C; and urokinase (UK) - see reference D. References cited herein by reference are as follows: (A) Lottenberg, R., U. Christensen, C.M. Jachson &amp; P. L. Coleman; &quot; Coagulation Protease Experiment Using Peptide Chromogen and Fluorogenic Substrates, Assay of Coagulation Proteases Using Peptide Chromogen and Fluoroge 49 * 1 5 10 15 65432 Case 4514?

Mod. 71-20,000 ex. - 90/08 20 25 nic Substrates) &quot;, Methods in Enzymology, (Methods in Enzyme-Logy) Vol. 80; Academic Press, New York, NY (1981), pp. 341-361; (B) Geiger, R. &quot; Kalicreine &quot; (Kallikrein), Methods for Enzymatic Analysis, (Methods of Enzymatic Analysis), Vol. V, 3rd edition, Bergmeyer, Academic Press, New York, N.Y. (1984), pp. 129 - 143; (C), Morris, J.P., S. Blatt, J.R Powell; D.K. Strickland &amp; F.J. Castellino, &quot; The Role of Lysine Ligation Regions in the Quinine Properties of Human Plasmine &quot; (Role of Lysine Binding Regions in the Kinetic Properties of Human Plasmin), Biochemistry (Biochemistry) Vol. 20, Ne 17 (August 18, 1981), p. 481X; (D) Wohl, R.C.L. Summaria &amp; K.C. Robbins, &quot; Kinetics in the Activation of Plasmid Human Nogenesis through Different Activator Species at pH 7.4 and 372C, (Kinetics of Activation of Human Plasminogen by Different Activator Species at pH 7.4 and 372C), Journal of Biological Chemistry, (The Journal Biological Chemistry, Vol. 255, N25 (March 10, 1980), 2005-2013; (E) Okunishi, H., J. Burton & J. Spragg. &Quot; A specificity of analog inhibitors of human urinary kallikrein substrates) &quot; Specificity of Substrate Analogue Inhibitors of Human Urinary Kallikrein), Hypertension, (Hypertension) Vol. 7, N23 Supp. 1 (May-June 1985), pp. 1-72-175; (F) Amundsen, E .; J Putter, P. Friberger, M. Knos, M. Larsbraten & G. Claeson, &quot; Methods for determination of Glandular kallikrein via chromogenic tripeptide substrate &quot; (Methods for the Determination of Glandular Kallikrein by Means of a Chromogenic Tripeptide Substrate), in Kinins-II Part A, Fuji, S., et al, eds, Plenum Press, New York, (1979), pp. 83-95. Another useful experience in the activity is based on a whole for determination of inhibition of slow binding enzyme described in Imperiali, B. & Inhibition of Inhibition Prohibited by Fluoromethyl Peptidyl Ketones (Inhibition of Serine Proteases by Peptidyl Fluoromethyl Keters), Biochemistry, (Biochemistry) Vol. 25, (1986), pp. 3760-3767. The method is modified as described below. 50 30 è 65432 Case 4514 c 1 5 10 15

I u

Mod. 71 20 25 is prepared in order to study the inhibition of slow binding of human plasmin (A #) and kallikrein (porcine pancreatic) (B +): Reactions: (A *) the reaction mixtures contain 78 mM Tris-HCl ), pH 7.4, 78mM NaCl, 0.2 mg / ml bovine serum albumin, 0.2mM S-2251 (D-Val-Leu-Arg-p-nitrolide), 0.5 U / ml plasmin 1 M) and varying concentrations of the test compound to be studied in a total volume of 1 ml. The stock solution of p-nitroaniline plasmin on enzymatic cleavage of S-2251 is monitored through a HP-8450 spectrophotometer system 400-410, programmed to measure the temperature at 30 ° C. Calculations: K,, and V (firm state inhibited rate) are determined by adapting the progressive curve (first 20 minutes) to the integrated rate equation (i) using &quot; software &quot; Labtech Notebook. The estimated is calculated for each Vg path and the uninhibited rate v (equation ii), with (S) = 0.2 mM and Km = 0.77 mM> (1 + (S) / Km) (1) (1) (1) (1) (1) (1) ) A test compound concentration pool vs kQbsd is then adapted for the different paths to one line, y = mx + b (see equation iii); KQn = m (1 + (S) / Km) and = b are calculated.

Finally it is calculated from the equation iv. (iii) K / K / K obsd off on There are several in vitro experiments that predict the anti-inflammatory activity of the compounds. Such in vitro experiments are useful in measuring the activity of the compounds of this invention. Such in vitro experiments are described in the following references, which are hereby incorporated by reference: Winter C.A., E.A., Risley, G.V.Nuss, &quot; Carrageenan induced edema in pulps of injured rats as an anti-inflammatory drug experiment &quot; (Carrageenan-Induced Edema 51, 65432 Case 4514,

1 5 10 15

Mod. 71 · 20,000 ex. - 90/08 in Hind Paw of the Rat as an Assay for Anti-Inflammatory Drugs Proc. Soc Exp Biol. N.Y., Vol. 111 (1962), p. 544-547; Vande, C. &quot; S. Margolin, &quot; Analgesic Theses Based on Acute Abdominal Pain Experimentally Induced in Rats &quot; (Federal Procedures-Vol. 15, (1956), p.494; Blackam, A., JW Burns, JB Farmer, HH Radziwonik, J. Westxick, , "A x-ray analysis of adjuvant arthritis in the rat." The effect of prednisolone and indo-metacine (An X-Ray Analysis of Adjuvant Arthritis in the Rat, The Effect of Prednisolone and Indomethacin), Agents and Actions, And Actions) Vol 7/1 (1977), p.145-151, Winter, CA & GW Nuss, &quot; Treatment of Adjuvant Arthritis in Rats with Anti-Inflammatory Drugs &quot; (Treatment of Adjuvant Arthritis in Rats with Anti-Inflammatory Drugs) Arthritis and Rheumatism Vol. 9, N-3 (June 1966), pp. 394-404; and Francis, MD, K.Hovancik &amp; RWBoycef &quot; NE-58095: a diphosphonate which avoids bone erosion and concludes the architecture of the joints in experimental arthritis "(NE-58095: A Diphosphonate Which Prevents Bone Erosion and Pre serves Joint Architecture in Experimental Arthritis), Int. J. Tiss. Reac., Vol. XI, N25, (1989), pp. 239-252.

COMPOSITIONS AND METHODS OF USE OF COMPOUNDS 25

The following non-limiting examples demonstrate contemplated compositions and uses of this invention. Tablets made by conventional procedures, each having the following composition: COMPONENTS QUANTITY (mg)

Ipoc-Dt-Bug-Phe-Arg-CFg Microcrystalline Cellulose 400 52 200 30 * 65432 Case 4514% Ρώ'Ι1ΐΐ83, - Pregelatinized Starch 200 Providone K-30 40 Magnesium Stearate 20 Each tablet a day to a patient due to arthritis. is given orally to alleviate inflammation 4 times by joints

Example 78

A lotion is made by the conventional procedure, having the lotion the following composition:

COMPONENTS

Adoc-D-Val-Phe-Arg-H glycerin paraben methyl paraben propyl carbopol 934 NaOH palmitate stearyl cetyl stearic acid fatty acids lanolin alcohol cetyl gum zantam sodium stearoyl-2-lactolate myristate isopropyl water

One gram of QUANTITY (%) 2.5 4.0 0.2 0.1 0.15 0.46 1.0 0.5 0.5 3.0 0.31 0.75 2.0 q.s. topically on the skin 53 65432 Case 4514

area of the burn, twice a day to reduce inflammation and pain.

Example 79

It is a solution made by conventional means, every 2 ml of the solution have the following composition:

COMPONENTS QUANTITY (mg) eBroc-D-t-Bug-Phe-Arg-H benzalkonium chloride sterile aqueous saline 80 40 q.s.

A dose of 2 ml of the solution is injected intramuscularly into a patient with arthritis to reduce inflammation and pain.

Example 80

It is a solution made by conventional means having the following composition: COMPONENTS Adoc-D-t-Bug-Phe-Arg-H benzalkonium chloride carboxymethyl cellulose sodium aqueous saline QUANTITY (%) 5.0 0.02 0.01 q.s.

A 2 ml dose of the solution is administered by inhalation to a patient as needed to relieve respiratory distress due to asthma. Notwithstanding the description of various embodiments of the invention, it is obvious to those skilled in the art that there are various changes in the invention which may be made without departing from the scope of the invention. - \\ 5/1 tion. The claims are intended to encompass all such changes. 5 Lisbon, 2.0 J1M. 1993 By THE PROCTER & GAMBLE COMPANY 10 LO 80 / 06- fc 'V s £ i 20 VASCO MAKQUES LtlL? Agt., Vol. 18, No. 3, pp. 35-40.

Mod. 71-20,000 e *. - 90/08 55

Claims (4)

10 Mod. 71-20,000 ex. - 90/08 20 25 30 65432 Case 4514 1. A compound, and pharmaceutically acceptable salts thereof, for the manufacture of a medicament for the treatment of inflammations or pains having the structure: ## STR2 ## (O) -NH-CH-C (O) -Y wherein: (a) n is an integer between 0 and 2 ; (b) -R is selected from linear or branched alkyl, saturated or unsaturated with 1 or 2 double bonds, having 1 to 6 carbon atoms, and cyclic alkyl, saturated or unsaturated, saturated with 1 or 2 double bonds, having 3 and 13 carbon atoms; and the carbon atom to which -R is attached has a D or L configuration; (c) -R 'is selected from branched alkyl, saturated or unsaturated with 1 or 2 double bonds, having from 3 to 6 carbon atoms; saturated or unsaturated cyclic alkyl having 1 or 2 double bonds having from 3 to 13 carbon atoms and arylalkyl, the alkyl portion being saturated and having 1 to 3 carbon atoms; and the carbon atom to which -R 'is attached has an L-configuration; (d) -R &quot; (CBL) -A-NH9 or - (CH9) -A-B-Z-Z-III-354 65432 Case 4514 / Mod. Wherein m is an integer from 1 to 5, -A- is a covalent bond or-p-phenyl or p-cyclohexyl, and -B- is a covalent bond or -NH-; and the carbon atom to which -R &quot; is attached has an L-configuration; (e) -Y is hydrogen or trifluoromethyl; (f) -Z- is -O- or -NH-; (g) -v- is selected from -OC (O) -, -N (Q) C (O) -, -N (Q) C (S) -, -C (O) -, -SO2 and P 0) (OH) -; when -V- is -OC (O) -, -Z- is -NH-; (h) -X- is selected from cyclic alkyl, branched alkyl having at least 2 branching, and aryl, each having 5 to 20 carbon atoms; except when -V- is -OC (O) -, n is 0 and Y is -Η, -X is other than t-butyl and (i) -Q is selected from hydrogen; and straight chain or branched alkyl, saturated or unsaturated with 1 or 2 double bonds, having 1 to 6 carbon atoms; or -Q and -X are covalently linked to form a cyclic moiety which includes the nitrogen to which -Q is attached and from 5 to 20 carbon atoms. A compound according to claim 1, wherein the alkyl moieties of -R, -R 'and X are saturated, the alkyl portion of any alkylaryl being saturated and having 1 to 3 carbon atoms, all unsubstituted alkyl R 'moieties. Compounds according to claim 1 or 2, characterized in that -V- is selected from -O C (O) -, - N (Q) C (O) -, -N Q) C (S) -, and -C (O) -, preferably -OC (O) - and -NHC (O) -; all aryl moieties of -R and R 'are unsubstituted; n is 0 or 1; and all the alkyl and aryl moieties of -X are substituted. A compound according to any one of claims 1-3, wherein -Q is hydrogen; the carbon to which -R is attached has a D configuration; -Z- is -NH-; and -A- is p-phenyl or p-cyclohexyl and m is 1, or -A- is a covalent bond, -B- is -NH- and m is 2-4. Mod. 71-20,000 ex. A compound according to any of claims 1-4 wherein X is alkyl or cyclic aryl of carbon, and Y is preferably -CF 3. The compound, and pharmaceutically acceptable salts and hydrates thereof, having the structure: â € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒ- (A) -AA- is selected from butylglycyl, valyl, isoleucyl, leucyl, cyclohexylglycyl, cyclohexylalanyl, phenylalanyl, naphthylalanyl, tryptophyl and adamantylglycyl; 2 (b) -AA - is selected from phenylalanyl, naphthylalanyl, cyclohexylalanyl, leucyl and isoleucyl; (c) -AA - is selected from arginyl, lysyl and p-guanidino-phenylalanyl, preferably arginyl; (D) -X 'is selected from 7-butyloxycarbonyl, adamantyloxycarbonyl, adamantyl, fluorenyl, methoxycarbonyl, adamanteneacetyl, adamantylaminocarbonyl, morpholinoyl, noradamantyl, homoadamantyloxycarbonyl, isopinocanphanyl-xycarbonyl, 3,5-dimethyladamantyloxycarbonyl, endo-butyloxycarbonyl, naphthyloxycarbonyl and methyloxycarbonyl; and (e) -Y is hydrogen or trifluoromethyl; except when -Y is hydrogen, -X 'is other than | t-butyloxycarbonyl. A compound according to claim 6, characterized in that: 71-20,000 β *. (B) -AA- is phenylalanyl; (c) -AA- is arginyl; and (d) -X 'is selected from t-butylglycyl, valyl and isoleucyl; The compound of the present invention comprises a compound of the general formulas (I) and the pharmaceutically acceptable salts thereof, and the compound of the formula: ## STR1 ## wherein R 1, R 2 and R 3 are independently selected from the group consisting of: pharmaceutically acceptable salts and hydrates according to claims 1-7, having the structure: â € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒ Mod. 71 -20,000 e *. R &quot; R &quot; (O) -NH-CH-C (O) -Y wherein (a) n is an integer from 0 to 2; b) -R is selected from linear or branched alkyl, saturated or unsaturated, having 1 or 2 double bonds, having 1 to 6 carbon atoms, and saturated or unsaturated cyclic alkyl with 1 or 2 bonds (c) -R 'is selected from branched alkyl, saturated or unsaturated, with 1 or 2 carbon atoms, and the carbon atom to which -R is attached has a D or L configuration; 2-double bonds having from 3 to 6 carbon atoms; cyclic, saturated or unsaturated alkyl having 1 or 2 double bonds having 3 to 13 carbon atoms, and arylalkyl, the alkyl portion being saturated and having 1 to 6 carbon atoms; (D) -R &quot; is - (CH2) m-NH2 or - (CH2) m ABC (NH2) = = NH, in which R3 is a hydrogen atom; m is an integer between 1 and 5, -A- is a covalent bond, or p-phenyl or p-cyclohexyl, and -B- is a covalent bond or -NH-; and the carbon atom to which -R &quot; is connected has a configuration in L; (e) -Y is hydrogen or trifluoromethyl; (f) -Z- is -O- or -NH-; (g) -V- is selected from -OC (O) -, -N (Q) C (O) -, 35-54 Mod. 71-20-000 e *. (S) -, -C (O) -, -SO2 - and -P (O) (OH) -; when -V- is -OC (O) -, -Z-, is -NH-; (h) -X is selected from cyclic alkyl, branched alkyl having at least 2 branching, and aryl each having 5 to 20 carbon atoms; and (i) -Q is selected from hydrogen and saturated or unsaturated straight or branched chain alkyl having 1 or 2 double bonds having 1 to 6 carbon atoms; or -Q and -X are covalently linked, forming a cyclic moiety, which includes the nitrogen to which -Q is bound between 5 and 20 carbon atoms; and (2) a pharmaceutically acceptable carrier. Lisbon, By THE PROCTER &amp; GAMBLE COMPANY VC ^ MARQUES Aqç, re Cíí Ίί Ι Ί Ί Ί tv · · · Cô Cô Cô Cô Cô Cô Cô Cô Cô Cô Conce Conce Conce Conce Conce Conce Conce Conce Conce Conce Conce Conce Conce Conce Conce Conce Conce Conce Conce Conce 6-