LT3284B - Fibrinogen receptor antagonists - Google Patents

Abstract

We analyse the fibrinogen receptor antagonists, whose formula:<IMAGE>they are used for inhibiting fastening of fibrinogen with thrombocytes and for inhibiting thrombocyte aggregation.

Description

The present invention relates to the discovery and use of fibrinogen receptor antagonists of formula I for inhibiting fibrinogen binding to platelets and inhibiting platelet aggregation in mammals, generally humans.

Platelet interactions with the coagulative and fibrinolytic systems during maintenance of haemostasis may become pathogenic and may be advisable to prevent or require treatment. The fibrinogen receptor antagonists described in Formula I are useful in the treatment of various diseases associated with platelet aggregation and fibrin formation.

Interest in platelet inhibitors has increased again in connection with a better understanding of the role of platelets and thrombosis in vascular pathogenesis, including unstable angina, acute myocardial infarction and stroke.

Platelets are similar to non-nuclear cells found in the blood of all mammals involved in blood coagulation. Fibrinogen is a glycoprotein that is incorporated into the plasma. Fibrinogen is involved in platelet aggregation and fibrin formation during blood coagulation. Platelets settle on vascular microvasculature, where many physiological antagonists act, which initiate platelet aggregation that results in platelet formation that limits blood loss. If the platelet is blocked inside the blood vessels, normal blood circulation is impaired.

Platelet membrane receptors play a key role in platelet adhesion and aggregation processes. The interaction of fibrinogen with the Ilb / IIIa receptor on the platelet membrane complex is known to be essential for normal platelet function.

Zimmerman et al., U.S. Pat. No. 4,683,291 describes peptides used in the assay of fibrinogen-platelets, platelets-platelets and extracellular interactions. The peptides described were useful in stopping or preventing the formation of a blood clot or a blood clot.

Piershabache et al., U.S. Pat. 4,589,881 discloses a sequence of an 11.5 kDa polypeptide fibronectin moiety that confers activity to fibronectin.

Ruoslahti in U.S. Pat. No. 4,614,517 discloses a tetrapeptide that modifies cell adhesive activity on a variety of substrates; 1 provides polypeptides synthesized by Ruoslahti et al. To identify the smallest peptide with activity on cell adhesive activity. Ruoslahti et al. U.S. Pat. 4,578,078 describes analogous tetrapeptides.

Piershabacher et al., Proc. Natl. Acad. Sci USA, vol. 81, p. 5985-5988, October, 1984, disclose variants of a fibronectin cell recognition center that retain binding-promoting activity. Piershabacher et al. further investigated cell adhesion promoting activity in a number of structures close to the Arg-GlyAsp-Ser peptide and demonstrated that the amino acid residues of arginine, glycine and aspartate cannot be substituted even with similar amino acids, but that serine can be substituted with several amino acids without losing activity.

Ruoslahti et al., Science, Vol. 238, pp. 491-497, October 23, 1987, discusses cell adhesion proteins. They separately emphasize that sequencing the amino acid sequence of fibronectin binding to the cell domain and its duplication with synthetic peptides allows the secretion of the Arg-Gly-Asp (GRD) sequence as an essential structure that cells recognize as fibronectin.

Cheresh, Proc. Natl. Acad. Sci. USA, vol. 84, p. No. 64716475, September 1987, describes an Arg-Gly-Asp specific adhesion receptor involved in binding fibrinogen and von Willebrandt factor.

Adams et al. U.S. Pat. No. 4,857,508 describes tetrapeptides which inhibit platelet aggregation and thrombus formation.

Therefore, it is an object of this patent to provide fibrinogen receptor antagonists which are useful for inhibiting fibrinogen binding to platelets and inhibiting platelet aggregation. In another aspect, this patent relates to the provision of novel compounds, fibrinogen receptor antagonists. Other objects of this patent include the provision of methods for binding fibrinogen to platelets and platelet aggregation using novel agents as fibrinogen receptor antagonists. These and other objectives are accomplished in the ways described below.

The present invention provides compounds which are fibrinogen receptor antagonists of the formula:

O ft

The V .. τ

X- (CK) mY- 'CH ₂ i kC-NH-CK-CH-z.

R ¹ used in fibrinogen binding to platelets, and inhibition of platelet aggregation. The aforementioned materials can be used to effect the fibrinogen receptor using a method which comprises administering to a mammal, generally a human, a therapeutically effective but non-toxic amount of such compound.

A pharmaceutical composition comprising a suitable pharmaceutical excipient and dispersed therein in an effective but non-toxic amount is another aspect of the present invention.

fibrinogen receptor antagonists, the formula is used in a method based on both platelet binding and platelet inhibition. The fibrinogen receptor provided in the present invention is illustrated by the formula:

The compounds are described as antagonists of fibrinogen I aggregation by the compounds k- (CH) _m -yu-C-NH-CH-CHCH / ro wherein

X is

NU

NH NH

-NR R ³ , -NH-C-NR ⁷ R ³ , -C-NR ⁷ R ³ , R ⁷ -CNHO--, ^or - ^D

H ₂ N-CH ₂

R ⁷ -

II

wherein A = N and B = -CH ₂ -, or A = -CH ~ and B = NR ⁷ ;

Z is CO ₂ R;

O lt

-P-OH i o

ii

-P-OH i

wherein R ¹⁰ is C ₁ . ₈ alkyl, aryl, aryl C _x _ ₈ alkyl;

u is -CH-, -C-, or -N-;

v is -CH-, -C-, or -N-;

R and R ¹ are independently hydrogen, aryl, where aryl is defined as a mono- or polycyclic aromatic system consisting of a 5 or 6 membered ring containing 0, 1, 2, 3 or 4 heteroatoms selected from nitrogen, oxygen and sulfur; and phenyl, substituted or unsubstituted with one or more groups selected from hydrogen, fluoro, chloro, bromo, iodo, cyano, trifluoromethyl, C _x . ₃ alkoxyl, C _x . ₅ alkylcarbonyloxy, C _ _{x 5} alkoxycarboxyl C _{first 5} alkyl, amino Ο _χ . ₅ alkyl, hydroxycarbonyl C _o _ ₅ alkyl, or hydroxycarbonyl C _x - ₅ alkoxy, C ₀ _ ₆ alkyl-, unsubstituted or substituted with one or more groups selected from fluoro, chloro, bromo, iodo, hydroxy, _{x. 5} alkylcarbonyl (C ₀ _ ₈ alkyl) amino, aryl C _x _ ₅ alkylcarbonyl (C ₀ _ ₈ alkyl) amino, aryloxy, Ci- ^ allTime koksilo B 3284 C _{x. 5} alkoxycarbonyl, C ₀ _ ₅ alkylaminocarbonyl, C _{x. 5} alkyl-carbonyloxy, C ₃ . ₈ cycloalkyl, aryl, oxo, amino, C ₁ . ₆ alkyl, C ₁ . ₃ alkylamino, aminoC ₁ . ₃ alkyl, arylC-5 alkilaminokarbinilo, fenilC _{x. 3} alkylamino, aminokarbonilC ₀ _ ₄ alkyl, C ₁ _ ₈ alkyl-sulfonyl (C _{0. 8} alkyl) amino, arylC _{o. xo} alkylsulphonylamino onil- (c _{0th 8} alkyl) alkylsulphonylamino onyl,

C _0-8 alkilosulf onyl, hydroxycarbonyl-C ₀ _ ₅ alkyl, C _{x. 8} alkoxycarbonyl (C _{0th 8} alkyl) amino, arylC ₀ - _X0 alkoxycarbonyl (C ₀ _ ₈ alkyl) amino, C _{0th 8} alkylamino carbonyl (C _{0th 8} alkyl) amino, arylC _{0th 8} alkylaminocarbonyl (C _{0th 8} alkyl) amino, C ₈ _ ₀ alkilaminokarboniloksilo, arylC _{0th 10} alkylaminocarbonyloxy, C ₀ . ₈ alkylsulfonylamino (C _{0th 8} alkyl) amino, arylC ₀ _ ₈ alkylsulfonylamino (C _{0th 8} alkyl) amino, C _{0th 8} alkylaminosulfonyl or arylC ₀ _ ₈ alkylsulfonylamino, provided that the carbon atom attached to R or R ¹ having only one hetero atom;

R ^{2 is} hydrogen, C ₁ . ₁₂ alkyl, substituted or unsubstituted with one or more C _{x 6} alkyl groups,

Oh it

R 'O

-CH2DC-R *

II

-CHOCR *

CH ₃ CH or

-C-O ·

I »O II

--- r.

g wherein R = branched or unbranched C _{x 6} alkyl, or 9 phenyl and wherein R when more than one may be the same or different;

3. 4

R, R and R are independently hydrogen, C _x . ₁₂ alkyl substituted or unsubstituted with one or more C _{x 6} alkyl groups, arylC ₀ . ₄ alkyl, or cyano, provided that R ⁷ and R ³ are independently cyano, X is

-NH-C-NR ^ R ³ *

NH

II or -C-NR '''R ^: 3k is 1-4;

m is 1-4;

p is 1-6;

q is 0-2;

or salts or isomers which are acceptable to the family.

Preferred compounds of the present invention are the following formulas:

their optical have these which

R

R ⁷ -NNH, a

HjN-CH _{2 in} which

C-. -S (0) _q - or -O-;

Z is CO ₂ R ² Y is

O

or wherein n is 1, 2 or 3;

R and R ¹ are independently selected from a phenyl, thiophene, imidazole, naphthyl, indole, indazole, tionafteno from substituted or unsubstituted hydroxycarbonyl C ₀ _ ₅ alkyl, hydroxy, halogen, C ₁ _ ₃ alkyl unsubstituted or substituted by one or more groups, selected from aryl, aryloxyl, C ₁ . ₁₀ alkoxyl, C ₀ . ₅ alkylaminocarbonyl, arylC _0-5 alkylaminocarbonyl, hydrogen, C _{0th 6} alkyl substituted or unsubstituted by one or more groups selected from halogen, hydroxyl, C _x 1 _ ₆ alki sul f arrested veneer arylC _{0th 6} alkylsulphonylamino Onil amino Cį.galkilsulf onyl, arylC ₀ _ ₆ alkylsulphonyl, C _x _ ₆ alkylcarbonylamino, arylC ₁ _ ₅ alkylcarbonylamino, aryloxy, C _x _ ₁₀ alkoxy, C _x _5alkoksikarbonilo C ₀ _ ₅ alkylaminocarbonyl, C _{x. 5} alkylcarbonyloxy, C ₃ _ ₈ cycloalkyl, aryl, oxo, amino, _{first 6} alkyl, C ₁ . ₃ alkylamino, aminoC ₁ _ ₃ alkyl, arylC ₀ _ ₅ alkylaminocarbonyl fenilC _x _ ₃ alkylamino, aminokarbonilC _{0th 4} alkyl, or hidroksikarbonilC ₀ _ ₅ alkyl, with the proviso that the carbon atom is connected to R or R ¹ has only one hetero atom,

R ² is hydrogen, ε _{χ 12} alkyl, substituted or unsubstituted with one or more C _{x 6} alkyl groups,

O! L

-CK ^ .GC-R **,

R ”O I II

-CKOCR * ''

t) i

-GG

Ft II '* where R is a branched or unbranched C ₁ _ ₆ alkyl, or phenyl, and wherein R ^9, when there is more than one, can be the same or different;

R, R and R are independently hydrogen or C ^ alkyl, substituted or unsubstituted by one or more C _x _ ₆ alkyl groups;

k is 1-4;

m is 1-4;

q is 0 or 2;

p is 1-3;

or pharmaceutically acceptable salts thereof, or optical isomers thereof.

The preferred compounds of the present invention have the following formulas:

X is

\ or where d = -0-S- or -CZ is CO ₂ R '

Y is

R

R

O <- * f tj ėJ

Wherein n is 1, 2 or 3;

R and R ¹ are independently selected from phenyl, thiophene, imidazole, naphthyl, indole, indazole, thionaphthene from hydroxy, halogen, C 1-4 alkyl, substituted or unsubstituted hydroxycarbonyl C ₀ . ₅ alkyl, unsubstituted by one or more groups selected from aryl, aryloxy, C _{first 10} alkoxyl, C ₀ . ₅ alkylaminocarbonyl, arylC ₀ . ₅ alkylaminocarbonyl, hydrogen, C ₀ . ₆ alkyl substituted or unsubstituted with one or more groups selected from halogen, hydroxyl, C ₁ . ₅ alkylsulfonylamino, arylC 1-6 alkylcarbonylamino, aryloxy, C _1-10 alkoxyl, C 1-4 alkoxycarbonyl, C ₀ . ₅ alkylaminocarbonyl, C ₁ _ ₅ alkylcarbonyloxy, C ₃ _ ₈ cycloalkyl, aryl, oxo, amino, C _{x. 6} alkyl, C ₁ _ ₃ alkylamino, aminoC ₁ _ ₃ alkyl, arylC ₀ _ ₅ alkylaminocarbonyl, phenyl C _{first 3} alkylamino, aminocarbonylC ₀ . ₄ alkyl, or hidroksikarbonilC ₀ _ ₅ alkyl, with the proviso that the carbon atom is connected to R or R ¹ has only one hetero atom,

R ² is hydrogen;

R ⁷ , R ³ and R ⁴ are hydrogen;

m is 1-4;

p is 2-4;

or pharmaceutically acceptable salts thereof, or optical isomers thereof.

The following abbreviations are used in the present invention:

Bn, benzyl; MMM, N-methylmorpholine; HOBt, 1-hydroxybenzotriazole; EDC, 1- (3-dimethylaminopropyl) 3-ethylcarbodiimide hydrochloride; DMF, dimethylformamideLT 3284 B das; BOC, t-butyloxycarbonyl; pTSA, p-toluenesulfonic acid; DMS, dimethylsulfide; TFA, trifluoacetic acid; THF, tetrahydrofuran; TBDMS, t-butyldimethylsilyl.

Pharmaceutically acceptable salts of the compound of formula I include the conventional non-toxic salts of the compound of formula I or the quaternary ammonium salts, for example, of non-toxic inorganic and organic acids. For example, such common non-toxic salts include those derived from inorganic acids such as salt, bromine, sulfur, sulfamine, phosphorus, nitrogen, and the like; and salts derived from organic acids such as acetic, propane, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic, maleic, hydromalein, phenylacetic, glutamine, benzoic, salicic, sulfanyl, 2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isethionic and the like.

Pharmaceutically acceptable salts of the present invention may be synthesized from a compound of formula I or an acidic residue. In general, the salts were bases or acids of the formula having a base by known chemical methods obtained by reacting free stoichiometric amounts or an excess of inorganic or organic acid or base the salt is to be obtained in a suitable solvent or a mixture of various solvents.

The pharmaceutically acceptable salts of the salts of formula I are readily obtainable by the customary reaction of Formula I acids with an appropriate amount of a base such as alkali or alkaline earth metal hydroxides such as sodium, potassium, lithium, calcium or magnesium, or organic bases such as such as an amine such as triethylamine, piperidine, and the like, or quaternary compounds such as tetramethylammonium dibenzylethylenediamine, pyrrolidine, benzylamine, with ammonium hydroxide, hydroxide and the like.

The compounds of the formula are used to inhibit fibrinogen binding to thromibocytes, inhibition of platelet aggregation, treatment of emerging thrombi or emboli, and as a preventive agent against thrombus or emboli formation, and as an inhibitor of cell fusion processes such as sperm and ovarian fusion, . These compounds are useful as pharmaceutical agents for mammals, especially humans. The compounds of the present invention can be used in patients who are desirable for thrombotic protection by inhibiting the binding of fibrinogen to the Ilb / IIIa receptor on the platelet membrane glycoprotein complex. The compounds of the present invention may also be used to prevent or modulate the development of acute or chronic myocardial infarction, unstable angina and thrombolytic stroke. In addition, they may be useful in peripheral arterial surgery (arterial transplantation, carotid endarterectomy) and cardiovascular surgery, where external effects on the arteries and organs and / or platelet interactions with artificial surfaces lead to platelet aggregation and destruction. Aggregated platelets may form thrombus or thromboembolism. The compounds of the present invention may be used in surgical patients to prevent thrombus or thromboembolic events.

In cardiovascular surgery, extracorporeal circulation is usually used to saturate the blood with oxygen. Platelets adhere to the walls of an extraLT 3284 B corporate shunt. Adhesion depends on the interaction between GPIIb / IIIa on the platelet membrane surface 252: H, pp and fibrinogen absorbed on the shunt (Gluszko et al., Amer.J. Physiol. 1987,

615-621). Platelets released from artificial surfaces exhibit poorer hemostatic function. The compounds of the present invention may be used to prevent adhesion.

Other applications of these compounds include the prevention of platelet thrombosis, etromboembolism, reocclusion and restenosis during or after thrombolytic therapy, and the prevention of platelet thrombosis, thromboembolism, reocclusion and restenosis after coronary and other arterial angioplasty and after coronary artery bypass.

The compounds of Formula I may be administered to a mammal as a pharmaceutical composition which is effective as a standard pharmaceutical pharmaceutically acceptable, sometimes known non-toxic, and therapeutically effective amount, usually in association with carriers or diluents, adjuvants such as alum. The compounds may be administered orally or parenterally, including intravenous, intramuscular, intradermal, subcutaneous, subcutaneous and topical application.

For the use of oral fibrinogen antagonists, the compounds of the present invention may be used, for example, in the form of tablets or capsules, or in the form of an aqueous solution or suspension. For oral tablets, conventional excipients, including lactose and corn starch, and lubricating agents, such as magnesium stearate, are usually added. For oral administration in capsule form, useful diluents include lactose and dry corn starch. When aqueous suspensions are required for oral use, the active ingredient is mixed with emulsifying and suspending agents. If desired, sweetening and / or flavoring substances are added.

In the case of intramuscular, intradermal, intravenous and subcutaneous administration, sterile active ingredient buffer solutions are generally formulated to provide an appropriate pH. For intravenous administration, total dissolved solids should be monitored to obtain an isotonic solution.

The present invention also encompasses pharmaceutical compositions comprising a therapeutically effective but non-toxic amount of the active compound of Formula I, with or without a pharmaceutically acceptable excipient or diluent, for use in the treatment of disorders associated with platelet aggregation, fibrin formation, thrombus formation and embolism. and avoidance.

The compositions described in the present invention include the fibrinogen receptor antagonist compounds of the present invention together with pharmaceutically acceptable excipients, e.g., saline at pH 7.4, for use in the inhibition of platelet aggregation. These compositions may also be used in combination with anticoagulants such as heparin or warfarin. These compositions may also be used in combination with thrombolytic agents such as plasminogen activators or streptokinase to prevent platelet aggregation in a sharper form. The compositions may also be used in combination with antithrombotic agents such as aspirin. The compositions are soluble in aqueous solutions and can therefore be effectively applied in the form of solutions.

When using the compounds described in Formula I as fibrinogen antagonists for the treatment of humans, the daily dose is usually determined by the attending physician based on age, weight, and individual patient response and the severity of the patient's symptoms.

In one embodiment, an appropriate amount of a compound is administered orally to an infarct patient before or after angioplasty. Administration is done before or after angioplasty and corresponds to an amount sufficient to prevent platelet aggregation, e.g., an amount that achieves a steady state plasma concentration of about 0.01-100 μΜ, usually about 0.01-10 μΜ.

The present invention also encompasses a pharmaceutical composition comprising the compounds of the present invention in combination with tissue plasminogen factor activator or streptokinase. The present invention also encompasses a method of accelerating thrombus lysis and preventing reocclusion comprising administering to the patient an effective amount of the compositions described herein.

The present invention provides a method of inhibiting the binding of fibrinogen to blood platelets, inhibiting platelet aggregation, treating thrombus or emboli, and preventing thrombus or emboli formation in a mammal comprising administering to a therapeutically effective but non-toxic amount of the compounds described herein. with or without fillers or diluents.

The present invention further provides a method of inhibiting fibrinogen binding to blood platelets, inhibiting platelet aggregation, treating thrombus or embolism, and preventing thrombus or embolism, a therapeutically effective but non-toxic mammalian compound comprising thrombolytic plasmids of the present invention. such as in combination with tissue streptokinase, heparin or activator or anticoagulants such as warfarin or antithrombotic agents such as aspirin with or without pharmaceutically acceptable carriers or diluents.

The following compounds and their respective IC ₅₀ values (concentrations which inhibit aggregation by 50% as compared to controls without these compounds) illustrate the present invention.

The compound

IKso

HN

0.097 μΜ

0.031 μΜ

O

100 μΜ

0.037 μΜ

0.2 μΜ

7.8 μΜ

Ο

ΗΝ

0.011 μΜ

0.03 μΜ

0.18 μΜ

0.056 μΜ

0.017 μΜ

0.55 μΜ

ΗΝ

0.096 μΜ

0.14 μΜ

8.1 μΜ

0.84 μΜ

0.27 μΜ

0.028 μΜ

3.7 μΜ

0.83 μΜ

0.58 μΜ

HN

O 'o ch ₃

_x CO ₂ H

0.2 μΜ

0.39 μΜ

0.42 μΜ

CH,

.co ₂ h

3.1 μΜ

0.55 μΜ

The compounds of the present invention may be prepared according to one of the following general procedures:

SCHEME I

The aryl or alkyl alcohol is protected, if necessary, and is oxidized to the corresponding aldehyde using oxalyl chloride / DMSO, pyridine chlorochromate, or similar reagents. Oxidation is usually performed for 0.5 to 6 hours. halogen-containing hydrocarbons in solvents such as CH ₂ Cl ₂ at -78 ° C to room temperature. The resulting aldehyde reacts with carboalkoxymethylene phosphorane to give the required unsaturated ester. The reaction is usually carried out for 1-18 hours. halogen-containing hydrocarbon solvents such as CH ₂ Cl ₂ at 0-60 ° C. Treatment of this ester with a chiral amine such as R - (+) - methylbenzylamine affords a diastereomeric mixture of amines which can usually be separated by chromatography.

SCHEME

The amino alcohol is protected via N using BOC or similar suitable groups such as CBZ, FMOC and the like. and treatment with iodine yields an alkyl halide. Typically, iodine and Ph ₃ P are used for this purpose and the reaction is carried out for 5 min. up to 10 hours in aromatic solvents such as benzene or toluene together with halogenated hydrocarbons, at 0-50 ° C, treatment with NaN ₃ or similar azide-transferring reagents yields the alkyl azide which is catalytically reduced for 0.5-1 hours. alcohols (EtOH, CH ₃ OH) give the corresponding amine at room temperature.

SCHEME

Alkylation of cyclic amides or urea intermediates is carried out in the presence of suitable bases such as LiN (TMS) ₂ or simple amines such as EtN (i-Pr) ₂ and corresponding alkylating agents such as alkyl alkyl iodide or bromide in ethyl bromoacetate. The introduction of two substituents in the intermediate is obtained by repeating the deprotonation / alkylation procedure. The use of alkyl reagents for alkylation and subsequent oxidation of RuCl ₃ / NaO ₄ or KMnO ₄ or other suitable reagents yields the terminal carboxylic acid, which can be further coupled via an amide bond to the C-terminal chiral or non-chiral amines to afford the following intermediates. The coupling reaction is typically carried out in DMF or halogen-containing hydrocarbon solvents such as CH ₂ Cl ₂ using standard reagents such as DCC, EDC, isobutyl chloroformate and the like. The final deprotection of the intermediates usually consists of base ester hydrolysis and acid catalyzed.

catalyzing N-deprotection,

The compounds represented by Formula I are the reaction schemes outlined below.

are obtained according to compound 1,

3-Indolpropanol, product.

is a commercial Aldrich Chemical

SCHEMA 'But-tojSiCl. DMF Ή -imidazole

ACjO.

- d iine. · 11 i a iii i n ~ j pi r i Ji nasCHjClj.

, S-cJi ZiZob i c i k i o— i S. 4.0 3Link - 7 - en a s

tSiMi / Ou

I

Ac

a) ι_ι t: s a 11 1 uj ctilor icJasCH'Cl

X,. -7 ° C preliminary ° C

b) Ph ₃ PrC »CO ₂ Et

SCHEMA Continued

Compound 7a is from American Tokyo Kansei, Ine. commercial product.

SCHEME

doc ₂ o. dhf

BOC-N iodine, PhjP, imidazole, benzene

BOC- fi i

0% PdZC. for ethane from h,

DOC c-, O-NH,

2

SCHEME = 0 = 1

5

EDC. iiOUT. NEC ₃ , DI-F, 7a

BOC-II

6th

SCHEMA Continued

PA \ NH

dccr;

I NH

JI p --- CO ₂ H \ - / o

7th

TrA. CH _? CJj. ani _{2 &} i _a5 -1 5 ° C f

SCHEME

SCHEMA Continued

EOC-N \

O

SCHEMA Continued

SCHEME

-chlorovaler i 1 o chlori daę ,,

D> CF. 1JKM

THF. Naf <ThtS) ₂ -78 ° C to 0 ° C

RuClj. CC1 «. CH2Cl2. H ₂ O. hlaTO «

BGC-N

O

V-CO..H _K Y DMF. 7 e

SCHEMA Continued

TFZi, CH ₂ Cl ₂ . aūizola5

HN

SCHEME

e) 4-Chorobutyryl chloride.

DMF. N11M A

---—--- f iT Ph

b) NaMCTMS}. THF. -70 ° C ^e Cikl25

LDA. THF, -70 ° C. then

DOC-H 'Fh ^J5a (separately) \ -J

35b 'KO /' aOH '· ^Ll ' ™ ^Γ · - ⁷⁸ ° ⁰

U

DOC-O C ™

DOC-Jf ^ S o

36a

36b

SCHEMA Continued

38b

SCHEMA Continued

BOC-N

38a

7a

IN NaOH. ethanol

SCHEMA Continued

38b

EDC. HO9T, NEtj. DMF. 7 a

IN NaOit ethanol

TFA. CH, Cl, anisole at -15 ° C

b

SCHEME

Svern oxidation

L1N.

SCHEMA Continued

SCHEMA Continued

SCHEME

σ · f \ χ C | V /! ό · -σ \ i * 3 O L δ o β i o

Oh to.

M - u

UI t

XJ

E

O

L

S? o h 4;

z.

-H> ~ l

J 0 b

z.

U

υ r · o

υ υΗ

C3 m I

Oh no

⁵ £ _J U z- \ / 'Χ c JQ

O <3 I

Ci

SCHEME

H3 _a C - '\ ^ CO, CH, (Aldrich)

pi vai oi 1o chloride p ^ r / V

Et j N. THF:

NC Hj OO <* ιηι ₂ · ηοι yti ^ A ^ CChCHj oo

J2x'_

FCC _? . ClIjCiL CHC1,

HH

ItaUSOj.

CH3CN

TlCO-i-Prjri,.

i-PrjHEC.

C, CH,

to.-siK t: * 3) j. p: f

BOC.

O

-15 ° C to 25 ° C

KIV, H-'BT. prc ;.

6th

SCHEMA Continued

O

1H. Cl'.jOH

BOC K.

H O i Λχο, η

O

Cr jCOjK CHjClj

HN

SCHEME

ICI • Η, Ν-'νΧΟ, Κ s <

i z ubuii 1 chl orthoiric acid in ethyl acetate 4-sample ltitor + al in

Cbz-Cly, -15 C

C, c bz: η ι - '^^ ίίΗ ^ χχτο, Et 19

20% Pd (OIQ, AcOH. CII, Oh-CH,

IDAc · Η, Ν · - ^ ΐη «\ χ; ο, Εί

O

DOCH i

h- ° <5

Lif <T8G),

THF, -78 ^e C ū t ū a 1 i 1 o'c

LIOOII

EDC. ICDT. PHF 1- Pr, HEt

IOC11 o

CH, χΐη ΐ '-' Ύ <· ι * 'X - c o, e t d O

SCHEMA Continued

B3

Os O «. 1θ4

H O CH,

N NeOit CH 3 OH

DOCN

Et, 3111 Cf, CO, H. C1I, C1,

SCHEME a

o '* o

t) H ~. HMy-IMT), dilo broioid0 * ^c o '' o 89

DOCH

//

CHO

CbO./HalO* THF. ! I, O

DOC1I '

BOC H čiC t; t UH č \ i>

Jonss reagent // 'o

COjH

EDC. .Έ9Γ.

i <i-Pr), EC.

UHF

ΓΟΟΝ

1. n-BuLl. THF. -70C

2 io. -23 ° for 2 hours

»»

HCl i r ch, v O 'N NaOH

SCHEMA Continued

TFA / CH ₂ C1 ₂ «

HN

cn ₃

SCHEME

y n

n a!

rf fa o

X o

O tN o

m m

o «Π (N <\

I o

Γ4 'Z'

SCHEME

DOCN

O-1,

FhjP. I, imidazole

DOCII

ICO v_ / _ Ό1

PbjP

CH2Cl2. 00 °

H,

10% Pd / C Ec OAc o

II

DOCN

THF

PPhjI

102 * 3OCH

1. When. HSCl, CH, CH

2.000.0

105 "· O’eye,

OCH.

106

ΝαΗςΊΤ-Ε),

DrCH3CO3EC, DMF, O °

B0CN

NH's ^ COjEt

107

SCHEME Continued »07» H UrtOH CIUO1I

OCH

I ^ zCOjH »09

DOCN ry -'- O

r.oc. iciir. d.-, ειι <ι-ιτ), cu,

I

1E11.11, H 57

O c »»

-Pkh-A ^ cc.ei _ O 1 07

ICr, cn _{/ t} CH, Cl,

1 1

SCHEME jtbenzi 1 chlorf orn »i atas _} thf J i <i-Pr) jEC. -15 ° C

Cbz

-O

112

PPh, Zi mi dazoi aiy'l ^

Cbz-H r

3

ItotKTS-S),. ΡΜΓ.

O

Cbz-N

-i

LD, \ TI ”-» D ° C then ai i 1 □ broira yet.

O

115 | CC1CHjCII. I! ₂ O IRuCl,. h'alO ₄

SCHEMA Continued

HjOil HOAc, Pd {OH} ₂

O

1 8

O

II

X (PCOH) ₂

SCHEME

1 9

2r..Chcm Der. 76. 1019 (1943)

20th

a) SOClj, CH 3 OH

b) benz i 1 chl air-f or ini ait I. '(l-Fr) ₂ Et

CbzHN -o O ₂ CHj 121

LDA. THF. -78 ° C followed by allyl bromide

122 trifluoroacetic acid _{from the} anisole as CH.Cl3, O ^; c

CO.CH,

SCHEMA Continued

EDC. HOBT. DMF. N (i-Pr) ₂ Et, Bu

tri f luoroacetic · acid,! anisole, CH ₂ C1 ₂

HN

S

O _h / - ^ C0 ₂ H

28th

SCHEMA Continued

123 toluene at 70 ° C

[DMl ·. ΝαίίζΤΙ · 3) ₂ oo to 1_C

126

Preparation of 3- (Indol-3-yl) -propanol-tert-butyl-dimethylsilyl ether (2)

To a stirred solution of 3-indolopropanol 1 (15 g, 86 mmol), DMF (200 mL) and imidazole (12.8 g, 0.19 mol) at 0 ° C was added tert-butyldimethylsilyl chloride (14.2 g, 95 mmol) and refluxed. bath. After 20 hours, the reaction mixture was diluted with ether and then washed with H ₂ O (2 times) and brine, dried over MgSO ₄ , concentrated to give the silyl ether 2 (29 g) as an amber oil. TLC R _f = 0.54 (20% ethyl acetate / hexane);

NMR (CDCl ₃ ) δ 8.07 (pis, 1H), 7.77 (d, J = 7Hz, 1H),

7.49 (d, J = 7Hz, 1H), 7.33 (t, J = 7Hz, 1H), 7.26 (t, 7Hz, 1H), 7.12 (s, 1H), 3.84 (t, J = 6Hz, 2H), 2.95 (t, J = 7Hz, 2H), 2.08 (m, 2H), 1.08 (s, 9H), 0.25 (s, 3H), 0.22 (s, H).

Preparation of N-Acetyl-3 (Indol-3-yl) -propanol-tert-butyl-dimethylsilyl ether (3)

A solution of indole 2 (29 g, 86 mmol)

CH ₂ Cl ₂ (450 mL), 1,8-diazobicyclo (5.4.0] undec-7-ene (38 mL, 0.26 mol), 4-dimethylaminopyridine (1 g, 8.5 mol) and acetic anhydride (32 mL, 0.34) The reaction mixture was concentrated and then diluted with ether The ethereal solution was washed with H ₂ O, 5% KHSO ₄ and brine, dried over MgSO ₄ and concentrated by flash chromatography (silica, 5% ethyl acetate / hexane). ) to give product 3 (27 g) as a yellow oil PSC R _f = 0.54 (20% ethyl acetate / hexane).

Preparation of N-Acetyl-3- (indol-3-yl) propanol (4)

To a suspension of ether 3 (27 g, 81 mmol) in THF (270 mL) was added a 1: 1 solution of 1M n-Bu ₄ NF in THF (244 mL, 0.24 mmol) and AcOH (14 mL) with stirring at room temperature. , 0.24 mmol). After 20 or so. the reaction mixture was diluted with ether and then washed twice with H ₂ O, brine, dried over MgSO ₄ and concentrated to give a yellow crystalline alcohol 4 (19 g). TLC R _f = 0.35 (60% ethyl acetate / hexane);

1 H NMR (CDCl ₃ ) δ 8.42 (m, 1H), 7.36 d, J = 7Hz, 1H), 7.36 (t, J = 7Hz, 1H), 7.29 (t, J = 7Hz, 1H) 7.27 (7d, t, J = 7Hz, 1H), 7.22 (s, 1H), 3.76 (t, J = 7Hz, 2H), 2.82 (t, J = 7Hz, 2H), 2.61 (s, 3H), 2.00 (m, 2H) ).

Preparation of 5- (N-Acetyl-indol-3-yl) -pent-2-enoic acid ethyl ester (5)

To a stirred solution of oxalic chloride (11.3 mL, 0.13 mmol) in CH ₂ Cl ₂ (400 mL) was added dropwise dry DMSO (12.4 mL, 0.17 mol) at -78 ° C. After 5 minutes the gas ceased to separate and a solution of alcohol 4 (19 g, 87 mmol) in THF (40 mL) was added. After 30 min, NEt _{3 was} added to obtain a thick mass. Before adding (carbethoxymethylene) triphenylforane (33.3 g, 96 mmol), the cooling bath is disconnected and stirred for 15 min. After 2 h, the reaction mixture was diluted with ether and then washed twice with H ₂ O, 5% KHSO ₄ , brine, dried over MgSO _4, and concentrated. Flash chromatography (20% ethyl acetate / hexane) gave a white solid (14 g), an olefin 5.

TLC R _f = 0.54 (60% ethyl acetate / hexane);

¹ H NMR (CDCl ₃ ) δ 8.42 (m, 1H), 7.50 d, J = 7 Hz, 1H), 7.34 (t, J = 7Hz, 1H), 7.28 (t, J = 7Hz, 1H) 7.19 (pis, 1H), 7.03 (dt, J = 18 and 7Hz, 1H), 5.88 (d, J = 18Hz, 1H), 4.19 (kv, J = 7Hz, 2H), 2.87 (t, J = 7Hz, 2H), 2.63 (m, 2H), 2.61 (s, 3H), 1.28 (t, J = 7Hz, 3H).

N- (R) -alpha-methyl-3 (R) - [2-indol-3-yl) ethyl-alanine ethyl ester (6a) and N- (R) -alpha-methylbenzyl-3 (S) - [2indole -3-yl) Preparation of β-alanine ethyl ester (6b)

A mixture of olefin 5 (2.77 g, 9.7 mmol) and R- (O-alpha-methylbenzylamine) (95.03 mL, 39 mmol) was heated in a refrigerator at 110 ° C for 40 hours. The cooled reaction mixture was applied to a flash column (silica, 40: 2: 1 Hexane: Ethyl acetate: 2-propanol) The (R, R) isomer (1.19 g) eluted first as a viscous yellow oil which solidified on storage Recrystallization from hexane gave crystalline material (R, S) Isomer 6b (1.55) g) eluting subsequently with a yellow oil containing approximately 10% (R, R) isomer 6a. 6a: R _f = 0.52 (60% ethyl acetate / hexane);

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.84 (pis, 1H), 7.52 (dd, J = 7.9, 0.7 Hz, 1H), 7.20-7.35 (m, 6H), 7.16 (tm, J = 7.1,

1.3 Hz, 1H), 7.08 (tm, J = 7.3, 1.1 Hz, 1H), 6.70 (pi d, J = 2.4, 1H), 4.10 (kv, J = 7.1 Hz, 2H), 3.90 (kv, J = 6.6 Hz, 1H), 2.80-2.90 (m, 2H), 2.68 (ΑΒΧ dt, J = 16, 7.9 Hz, 1H), 2.53 (ΑΒΧ dd, J = 14.5, 5.9 Hz, 1H), 2.42 (ΑΒΧ dd, J = 14.6, 5.3 Hz, 1H), 1.79 (kv, J = 7.5 Hz, 2H), 1.33 (d,

J = 6.4 Hz, 3H), 1.22 (t, J = 7.1 Hz, 3H).

6b: R _f = 0.42 (60% ethyl acetate / hexane);

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.95 (pis, 1H), 7.57 (dd,

J = 7.5, 0.7 Hz, 1H), 7.34 (dm, J = 8.1, 0.7 Hz, 1H), 7.177.30 (m), 7.11 (tm, J = 7.9, 0.9 Hz, 1H), 6.89 (pi d,

J = 2.2, 1H), 4.02-4.15 (ΑΒΧ m, 2H), 3.89 (kv, J = 6.6 Hz,

1H), 2.95 (m, 1H), 2.82 (δ ddd, J = 15, 9.7, 5.9 Hz,

1H), 2.69 (ΑΒΧ ddd, J = 15, 9.7, 6.0 Hz, 1H), 2.47 (ΑΒΧ dd, J = 15.0,

5.1 Hz, 1H), 2.40 (ΑΒΧ dd, J = 15.0, 7.7, 1H), 1.96 (m, 1H), 1.83 (m, 1H), 1.30 (d, J = 6.6 Hz, 3H), 1.21 (t, J = 7.1, 0.7 Hz, 3H).

Preparation of 3 (R) - [2-indol-3-yl) ethyl-alanine ethyl ester (7a)

Amine 6a (996 mg, 2.74 mmol) is dissolved in 10 mL of ethanol.Add PdOH ₂ C, 128 mg)

Pearlman's catalyst (20% flask is filled with hydrogen and pressure. After 16 hours, additional catalyst portion (122 mg) is added and hydrogen is added. After 4 hours, the sample is filtered through celite and concentrated to give amine 7a (707 mg, 99%, 95% purity). : R _f = 0.22 {10: 1 NH ₃ saturated

CHCl ₃ : EtOAC) ethyl acetate / hexane);

^Χ Η NMR (400 MHz, CDCl ₃ ) δ 8.01 (pi s, 1H), 7.60 dt, J (8.9, 04H ₂ , 1H), 7.35 (dt, J = 8.1, 0.9 Hz, 1H), 7.19 (td, J = 7.1, 1.3 Hz, 1H), 7.11 (td, J = 7.1, 1. 2 Hz, 1H), 6.99 (pi d, J = 2.2 Hz, 1H), 4 .15 (sq, J = 7.1, 2H), 3.72 (sq, J = 7.0, 1H), 3.29 (m, 1H) , 2.92-2.78 (m, 2H), 2.53 (ΑΒΧ dd, J = 15.6, 4.0 Hz, 1H), 2.33 (ΑΒΧ dd, J = 15  6, 8.8 Hz, 1H), 1.92-1.73 (m, 2H), 1.25 (sq, J = 7. 1 Hz, 3H).

Preparation of N-BOC-4-piperidinethanol (9)

To a stirred solution of 4-piperidinethanol 8 (18.7 g, 0.14 mol) in DMF (200 mL) at 0 ° C was added N-tert-butoxycarbonyl anhydride (31 g, 0.14 mol). After 1 or.

disconnect the cooling bath and stir the reaction mixture with water. The reaction mixture was diluted with ether and then washed twice with H ₂ O, brine, dried over MgSO _4, and concentrated to give 9 (26 g, 62) as a colorless oil. TLC R _f = 0.25 (40% ethyl acetate / hexane);

¹ H NMR (300 MHz, CDCl ₃ ) δ 4.09 (pis, 2H), 3.72 (t, J = 7Hz, 2H), 2.70 (m, 2H), 1.75-1.10 (m, 7H), 1.46 (s, 9H) ).

Preparation of N-BOC-4-piperidine ethyl azide (10)

A solution of 10 (5.0 g, 14.7 mmol), DMSO (75 mL) and NaN ₃ (1.9 g, 29.4 mmol) was heated for 2 h. at 70 ° C. The cooled reaction mixture was diluted with ethyl acetate and washed 2x with water and brine, dried over MgSO ₄ and concentrated to give 11 (3.6 g, (96%) as a colorless oil. PSC R _f = 0.75 (30% ethyl acetate / hexane);

¹ H NMR (300 MHz, CDCl ₃ ) δ 4.11 (m, 2H), 3.36 (t, J = 7Hz, 2H), 2.73 (m, 2H), 1.73 (m, 3H), 1.49 (s, 9H), 1.15 (m, 2H).

Preparation of N-BOC-4-piperidine ethylamine (12)

A mixture of 11 (1.1 g, 4.3 mmol), 10% Pd / C (0.16 g) and ethanol was stirred under a hydrogen atmosphere (1 atm) for 1.5 h. The reaction mixture is then filtered through a Celite filter and concentrated the filtrate to give crude 12 as an oil. TLC R _f = 0.18 ((9: 1: 1 CH ₂ CH ₂ / CH ₃ OH / HOAc);

¹ H NMR (300 MHz, CDCl ₃ ) δ 4.10 (m, 2H), 2.78 (t,

J = 7Hz, 2H), 2.70 (m, 2H), 1.80 (m, 2H), 1.67 (m, 2H),

1.52 (m, 1H), 1.47 (s, 9H), 1.17 (m, 2H).

Preparation of 1- [2- (N-BOC-4-piperidin-4-yl) ethyl] -3-propen-2-yl- (2-imidazolidinone) (14)

To a solution of 13 (1.5 g, 17 mmol) in DMF (75 mL) at room temperature was added LiN- (TMS) ₂ (1M in 17 mL hexane) until a precipitate formed. Allyl bromide (1.6 mL, 19 mmol) was then added to the reaction mixture. After 15 minutes LiN- (TMS) ₂ (14 mL) was added again to the reaction mixture, followed by 5 min. iodide 10 (5.9 g, 17 mmol) was added. The reaction mixture was stirred for 20 hours. and diluted with ether. The ether layer was washed twice with water and brine, dried over MgSO _4, and concentrated. Flash chromatography (40% ethyl acetate / hexane) gave 14 (0.8 g, 15%) as a colorless oil.

TLC R _f = 0.75 (70% ethylhexane);

¹ H NMR (300 MHz, CDCl ₃ ) δ 5.80 (m, 1H), 5.23 (m, 2H),

4.10 (m, 2H), 3.82 (m, 2H), 3.30 (m, 6H), 2.72 (m, 2H), 1.70 (m, 3H), 1.50 (m, 2H), 1.70 (m, 3H), 1.50 (m, 2H), 1.46 (s, 9H), 1.15 (m, 2H).

Preparation of 1- [2- (N-BOC-4-piperidin-4-yl) ethyl] -3-acetyl- (2-imidazolidinone) (15)

To a vigorously stirred solution of 14 (450 mg, 1.4 mmol) in CCl ₄ , acetonitrile and water was added RuCl ₃ (12 mg, 4.4 mol) and NaIO ₄ at room temperature. After 60 or so. the reaction mixture was filtered through a celite filter and washed with ethyl acetate. The filtrate is extracted with saturated NaHCO ₃ , the aqueous phase is acidified to pH 3 with 5% KHSO ₄ . The acidified aqueous phase is further extracted twice with ethyl acetate, the organic layer is dried over MgSO ₄ and concentrated. Flash chromatography (silica gel, CH ₂ C ₁₂ / CH ₃ OH / HOAc) gives 15 (60 mg, 12%) as a colorless oil.

TLC R _f = 0:29 (9.5: 0.5: 0.5 CH ₂ C ₁₂ / CH ₃ OH / HOAc);

^X H NMR (300 MHz, CDCl ₃ ) δ 4.05 (m, 2H), 3.90 (m, 2H), 3.45 (m, 2H), 3.40 (m, 2H), 3.24 (m, 2H), 2.69 (m, 2H), 1.72 (m, 2H) , 1.46 (m, 3H), 1.46 (s, 9H), 1.15 (m, 2H).

1- [2- (N-BOC-4-piperidin-4-yl) ethyl] -3-acetyl- (2-imidazolidinone) -3-acetyl-39 (R) - [2- (indol-3-yl) ethyl ] Preparation of β-alanine ethyl ester (16)

To a mixture of 15 (60 mg, 0.17 mmol), 7a (66 mg, 0.25 mmol), HOBT (30 mg, 0.22 mmol) and DMF (1.1 mL) at 0 ° C was added ESD (0.42 mg, 0.22 mmol) ), then disconnect the cooling bath. After 20 or so. the reaction mixture was diluted with ethyl acetate, washed with water, 5% KHSO ₄ solution, and the organic layer was dried and concentrated. After high-speed chromate (silica gel, ethyl acetate), the MgSO ₄ salt and the graphite were obtained (35 mg, 35%) as a colorless oil.

TLC R _f = 00:20 (ethyl acetate);

^X H NMR (300 MHz, CDCl; j) δ 8.07 (pis, 1H), 7.57 (d, J = 8 Hz, 1H), 7.37 (d, J = 8 Hz, 1H), 7.18 (t, J = 8 Hz, 1H), 7 .11 (t, J = 8 Hz, IH), 7.05 (pis, 1H), 6.89 (d, J = 9 Hz, 1H), 4.37 (m, IH), 4.13 (m, H), 4.13 (sq, J = 7 Hz, 2H), 4.07 (m, 2H) , 3.83 (dd, J = 18 Hz, 2H), 3.43-3. 20 (m, 6H), 2.80 (m, 2H), 2.56 (d, J = 5Hz, 2H), 2.00-1. 05 (m, 9H), 1.47 (s, 9H), 1.25 (t, J = 7Hz, 3H).

1- [2- (N-BOC-4-piperidin-4-yl) ethyl] -3-acetyl- (2-imidazolidinone) -3-acetyl-39 (R) - [2- (indol-3-yl) ethyl ] Preparation of βalanine (17)

A mixture of 16 (30 mg, 5.0 μ mol), IN NaOH (0.2 mL) and ethanol was stirred at room temperature for 1 h. The reaction mixture was diluted with ethyl acetate and 5% KHSO ₄ solution, the organic layer was washed with water, brine, dried over MgSO ₄ and concentrated to give 17 (30 mg, 100%) as a colorless oil.

TLC R _f = 0.74 (9: 1: 1 CH ₂ C ₁₂ / CH ₃ OH / HOAc).

1- [2-piperidin-4-yl) ethyl] -3-acetyl- (2-imidazolidinone) 3-acetyl-39 (R) - [2- (indol-3-yl) ethyl] β-alanine (18 ) receipt

To a solution of 17 (30 mg, 50 μπιοί), dichloromethane (300 μΐ) and anisole (15 μΐ, 100 μιηοΐ) was added TFA (0.3 mL) at -15 °. After 20 minutes the reaction mixture is concentrated and the remaining TFA is removed by distillation of the aceotropic mixture with toluene. Flash chromatography (silica gel, 10: 0.8: 0.8 methanol / NH ₄ / water) gives 18 (15 mg, 66%) as a white solid.

TLC R _f = 0.16 (10: 0.8: 0.8 methanol / NH ₄ OH / water);

¹ H NMR (300 MHz, CD ₃ OD) δ 7.44 (d, J = Hz, 1H), 7.20 (d,

j = 8: Hz, 1H), 6.95 (t, J = 8 Hz, 1H), 6.94 (s, 1H), 6.88 (t, J = 8 Hz, 1H) , 4. .17 (m, , 1H), 3.71 (s, 2H), 3.50- 3.10 (m, 8H) 2.86 (m, 2H), 2.70 (d, J = 6 Hz, 2H), 2.32 (m, 1H) , 1.90 (m, 4H), 1.55 (m, 1.H), 1.43 (m, 2H), 1.28 (m, 2H).

Preparation of 1- [2-N-BOC-piperidin-4-yl] ethyl] - (2-pyrrolidinone) (19)

To a solution of 12 (2.7 g, 11.8 mmol), acetonitrile (60 mL) and diisopropylethylamine (4.1 mL,

23.6 mmol) was added 4-chlorobutyryl chloride (2.6 mL, 23.6 mmol) with stirring at 0 °, followed by removal of the cooling bath. After 5 minutes the reaction mixture was diluted with ethyl acetate, washed twice with water, brine, and the organic layer was dried over MgSO ₄ and concentrated. The crude amide was dissolved (60 mL) cooled to 0 ° C and NaN (TMS) ₂ (1M in THF, 11.8 mL) was added. After 5 minutes the reaction mixture was diluted with ethyl acetate, washed with water and brine, and the organic layer was dried over MgSO ₄ and concentrated. Flash chromatography (silica gel, 70% ethyl acetate / hexane) gives 19 (0.4 g, 12%) as a colorless oil.

TLC R _f = 0.29 (70% ethyl acetate / hexane);

¹ H NMR (300 MHz, CDCl ₃ ) δ 4.10 (m, 2H), 3.39 (t, J = 7 Hz, 2H), 2.70 (m, 2H), 2.40 (m, 2H), 2.05 (m, 2H) , 1.72 (m, 2H), 1.46 (m, 3H), 1.45 (s, 9H), 1.13 (m, 2H).

Preparation of 1- [2-N-BOC-piperidin-4-yl) ethyl] -3-propen-2-yl (2-pyrrolidinone) (20)

To a solution of 19 (325 mg, 1.1 mmol) in THF (5 mL) was added dropwise LDA (0.5 M in 2.4 mL THF) at -78 ° C. After 15 minutes allyl bromide (0.16 ml,

2.2 mmol), the reaction mixture was washed at 78 ° C for 1 h. and quenched with HOAc (0.1 mL). The reaction mixture was diluted with ethyl acetate, washed with water, and brine, and the organic layer was dried (MgSO ₄ ) and concentrated. Flash chromatography (silica gel, 50% ethyl acetate / hexane) gave 20 (160 mg, 45%) as an oil.

TLC R _f = 0.23 (50% ethyl acetate / hexane);

NMR (300 MHz, CDCl ₃ ) δ 5.80 (m, 1H), 5.10 (m, 2H), 4. 1 (m, 2H), 3.32 (m, 4H), 2.70 (m, 2H), 2.56 (m, 2H), 2. 20 (m , 3H) , 1.73 (m, 3H), 1.48 (s, 9H), 1.13 (m, 2H).

- Preparation of [2-N-BOC-piperidin-4-yl) ethyl] -3-acetyl-2-pyrrolidinone (21)

Using the same procedure as for the 14 conversion to case 20, (130 mg, 0.4 mmol) was obtained by flash chromatography (silica gel, 9: 0.2: 0.2 CH ₂ C ₁₂ / CH ₃ OH / HOAc) 21 (80 mg, 48). in the form of an oil;

PSC R _f = 0.2 (9: 1: 1 CH ₂ Cl ₂ / CH ₃ OH / HOAc:

¹ H NMR (300 MHz, CDCl ₃ ) δ 4.05 (m, 1H), 3.40 (m, 4H),

2.96 (m, 1H), 2.83 (m, 1H), 2.70 (m, 2H), 2.55 (m, 1H), 2.40 (m, 1H), 1.90-1.10 (m, 7H), 1.48 (s, 9H) .

Preparation of [1- [2-N-BOC-piperidin-4-yl] ethyl] -2-pyrrolidone-3] -acetyl (2- (indol-3-yl) ethyl] β-alanine ethyl ester (22)

Using the same procedure as for conversion 15, 21 (120 mg, 0.35 mmol) were obtained by flash chromatography (silica gel, ethyl acetate) 22 (90 mg, 48) as an oil;

TLC R _f = 0.40 (ethyl acetate):

^Χ Η NMR (400 MHz, CDC1 ₃ ) δ 8.09 (bd, 1H), 7.58 (d, J = 8 Hz, 1H), 7.36 (d, J = 8 Hz, 1H), 7.17 (t, J = = 8Hz, 1H), 7.10 (t, J = 8 Hz, 1H), 7.07 (pis, 1H), 4.39 (m, 1H), 4.12 (sq, J = 7 Hz, 2H), 4.05 (m, 2H), 3.30 (m, 4H), 2.90-2.25 (m, 8H) , 2am - 1pm (m, 8H), 1.48 (s, 9H),

1.23 (t, J = 7Hz, 3H).

Preparation of [1- [2-N-BOC-piperidin-4-yl] ethyl] -2-pyrrolidone-3] -acetyl (2- (indol-3-yl) ethyl] β-alanine (23)

Using the same procedure as for the 16 conversion, 22 (80 mg, 0.14 mmol) were obtained in 23 (80 mg) analogous to the following reaction.

PSC R _f = 0.5 (9: 1: 1 CH ₂ Cl ₂ / CH ₃ OH / HOAc);

Preparation of [1- [2-piperidin-4-yl) ethyl] -2-pyrrolidone-3] -acetyl- (2 (indol-3-yl) ethyl] β-alanine (24)

Using the same procedure as in case 17 conversion, 24 (15 mg, 23%) of 23 (80 mg, 0.14 mmol) after flash chromatography (silica gel, 10: 0.65: 0.65 methanol / NH ₄ OH / water) were obtained.

PSC R _f = 0.53 (10: 1: 1 methanol / NH ₄ OH / water).

¹ H NMR (300 MHz, CDCl ₃ ) δ 7.47 (d, J = 7Hz, 1H), 7.25 (d,

J = 7 ' Hz, IH), 6.99 (m, 2H), 6.88 (t, J = 7Hz, IH), 4.19 (m, IH), 3.49 (m, 0.5 H), 3.40 (m, 0.5 H), 3.34- 2.95 (m, 6H), 2.90-2.60 (m, 4H), 2.48 (m, IH), 2.40- 2.20 (m, 4H), 2.13 (m, 1H), 2.00-1.05 (m, 11 H).

Preparation of 5-chloro-valeroylamino-2-propene (26)

To a mixture of allylamine 25 (2.0 g, 27 mmol), NMM (8.9 mL, 81 mmol) and acetonitrile (140 mL) was added 4-chloro-butyryl chloride (3.4 mL, 27 mmol) at room temperature. After 20 or so. the reaction mixture is concentrated. The residue was dissolved in ethyl acetate, washed with water, 5% KHSO ₄ , brine, dried over MgSO ₄ and concentrated to give 26 (4.1 g, 87%) as a yellow oil.

TLC R _f = 0.170 (40% ethyl acetate / hexane).

Preparation of 1- (propen-2-yl) -2-piperidone (27)

To a solution of 26 (2.0 g, 11 mmol) in THF (110 mL) was stirred Na-Na (TMS) ₂ (1M in THF, 11 mL) at -78 ° C and the cooling bath was removed. After 1.5 or. the reaction mixture was diluted with ethyl acetate, washed with water, 5% KHSO _4, and brine, dried over MgSO _4, and concentrated. Flash chromatography (silica gel, 60% ethyl acetate / hexane) gives 27 (1.1 g, 72%) as an oil.

TLC R _f = 0.15 (60% ethyl acetate / hexane);

¹ H NMR (300 MHz, CDCl ₃ ) δ 5.78 (m, 1H), 5.15, 4.00 (t,

J = 7Hz, 2H), 3.24 (m, 2H), 2.41 (m, 2H), 2.07 (m, 2H), 1.80 (m, 4H).

Preparation of 3- [2-N-BOC-piperidin-4-yl) ethyl] -1-pyrrolidone-3] -1-acetyl-2-piperidone (28)

To a solution of 27 (1.0 g, 7.2 mmol) in THF (70 mL) was added dropwise LDA (0.5 M, 15.8 mL) at -78 ° C. After 15 minutes a solution of iodide 10 (2.7 g, 7.9 mmol) in THF (5 mL) was added and the reaction mixture was slowly raised to -30 ° C over 1 h. The reaction was quenched by the addition of acetic acid (0.2 mL) and concentrated. Flash chromatography (silica gel, 40% ethyl acetate / hexane) gives 28 (1.5 g, 59%) as an oil.

TLC R _f = 00:58 (ethyl acetate);

⁷ H NMR (300 MHz, CDCl ₃ ) δ 5.77 (m, 1H), 5.15 (m, 1H),

4.06 (m, 2H), 4.00 (m, 2H), 3.23 (m, 2H), 2.59 (m, 2H),

2.19 (m, 1H), 2.05-1.25 (m, 11H), 1.48 (s, 9H),

1.10 (m, 2H).

Preparation of 3- [2-N-BOC-piperidin-4-yl) ethyl] -1-pyrrolidone-3] -3-acetyl-2-piperidone (29)

In a vigorously stirred mixture of 28 (0.84 g,

2.4 mmol), CCl ₄ (5 mL), acetonitrile (5 mL), NaIO ₄ (2.1 g, 9.8 mmol) and water (7.5 mL) at room temperature were added RuCl ₁ (50 mg, 0.2 mmol). After 20 or so. an additional portion of RuC1 ₃ (50 mg) was added. After 4 or. the reaction mixture was diluted with ethyl acetate and half-saturated NaHCO ₃ . After mixing and separating the layers, the aqueous phase is acidified with 5% KHSO ₄ and extracted with ethyl acetate. The organic layer was washed with brine, dried over MgSO _4, and concentrated. Flash chromatography (silica gel, 9: 0.2: 0.2 CH ₂ Cl ₂ / CH ₃ OH / HOAc) gives 29 (280 mg, 30%) as an oil.

TLC R _f = 0.36 (9.5: 0.5: 0.5 CH ₂ Cl ₂ / CH ₃ OH / HOAc).

[3- [2-N-BOC-piperidin-4-yl) ethyl] -2-imidazolidinone-1] acetyl [3 (R) - [(2- (indol-3-yl) ethyl] β-alanine Preparation of ethyl ester (30)

Using the same procedure as for conversion 15 to 16, of the 29 (280 mg, 0.76 mmol) after flash chromatography (silica gel, 85% ethyl acetate / hexane) gave 22 (300 mg, 64%) as an oil;

TLC R _f = 0.18 (85% ethyl acetate / hexane);

1 H NMR (300 MHz, CDCl ₃ ) δ 8.14 (m, 1H), 7.60 (d, J = 8 Hz,

1H), 7.39 (d, J = 8Hz, 1H), 7.20 (t, J = 8Hz, 1H),

7.13 (t, J = 8Hz, 1H), 7.05 (pis, 1H), 6.90 (d, J = 9Hz,

1H), 4.35 (m, 1H), 4.11 (g, J = 7Hz, 2H), 4.05 (m, 2H),

3.98 (s, 2H), 3.40 (m, 4H), 2.90-2.50 (m, 6H), 2.33 (m, 1H), 2.00-1.00 (m, 15H), 1.48 (s, 9H), 1.29 (9t,

J = 7 Hz, 3H).

[3- [2-N-BOC-piperidin-4-yl) ethyl] -2-imidazolidinone-1] acetyl [3 (R) - [(2- (indol-3-yl) ethyl] β-alanine receiving (30)

Using the same procedure as for the 16 conversion, 30 (160 mg, 0.26 mmol) were obtained (140 mg, 92%) as a white solid.

TLC R _f = 0.59 (9: 1: 1 CH ₂ Cl ₂ / methanol / HOAc).

Preparation of [3- [2-piperidin-4-yl) ethyl] -2-imidazolidinone-1] acetyl [3 (R) - [(2- (indol-3-yl) ethyl] β-alanine (32)

Using the same procedure as in case 17 conversion, 32 (55 mg, 47%) of 31 (140 mg, 0.24 mmol) after flash chromatography (silica gel, 10: 0.4: 0.4 methanol / NH ₄ OH / water) were obtained. in the form of a solid.

PSC R _f = 0.46 (10: 1: 1 methanol / NH ₄ OH / water);

¹ H NMR (300 MHz, CD ₃ OD) δ 7.54 (d, J = 8 Hz, 1H), 7.30 (d, J = 8 Hz, 1H), 7.03 (t, J = 8 Hz, 1H), 7.02 ( pls, IH),

6,976 (m, 1H), 4.2 Δ (m, 1H), 4.20 (d, J = 16 Hz, 0.5H), 4.12 (d, J = 16 Hz, 0.5H), 3.80 (d, J = 16Hz, 0.5H), 3.75 (d, J = 16 Hz, 0.5H), (s, 2H), 3.50-3.15 (m, 4H), 2.80 (m, 4H), 2.70 (d, J = 6Hz, 2H), 2.50-2.30 (m, 3H), 2.00- 1.20 (m, 15H).

Preparation of 3- [2-N-BOC-piperidin-4-yl) ethyl] -3-acetyl-2-pyrrolidinone (36a)

To a solution of benzylamine 35a (0.8 g, 2.2 mmol) in THF (20 mL) at -78 ° C was added 2 x freshly prepared lithium di-tert-butyl biphenylate (0.5 M in 10.8 mL). After hours quench the reaction with HOAc (0.1 mL) and concentrate. Flash chromatography (5% methanol / ethyl acetate) gave 36a (470 mg, 72%) as a white solid.

TLC R _f = 0.27 (10% methanol / ethyl acetate);

⁷ H NMR (300 MHz, C DC1 ₃ ) δ 6.03 (m, 1H), 4.10 (m, 1H), 3.35 (m, 2H), 2.70 (m, 2H), 2.33 (m, 2H), 2.70 (m, 2H), 1.92 (m, 1H), 1.80 (m, 1H), 1.70 (m, 2H), 1.50- -1.30 (m, 4H), 1.48 (s, 9H), 1.12 (m, 1H).

Preparation of methyl 1-acetate-3- [2-N-BOC-piperidin-4-yl) ethyl] -3-acetyl-2-pyrrolidinone (37a)

To a solution of 36a (340 mg, 1.1 mmol) in THF (10 mL) was stirred

At -78 ° C, NaN (TMS) ₂ (1M in 11 mL was added to the stream)

THF) and a removable cooling bath with ethyl bromoacetate (250 mL, warm 2 to -20 ° C, stop with HOAc (0.1 mL) and flash chromatography (silica gel, 50% ethyl acetate / hexane) to give 37a (380 mg, 90%) as an oil .

After 15 minutes 2 mmol) and After 2 hours, the reaction mixture is added to concentrate. After

TLC R _f = 0.66 (10% methanol / ethyl acetate);

¹ H NMR (300 MHz, CDCl ₃ ) δ 4.22 (q, J = 7Hz, 2H), 4.1 (m, 2H), 4.08 (s, 2H), 3.24 (m, 2H), 2.70 (m, 2H), 2.44 (m, 1H), 2.26 (m, 1H), 2.00-1.65 (m, 4H), 1.48 (s, 9H), 1.50-1.30 (m, 4H), 1.31 (t, J = 7Hz, 3H), 1.12 (m, 2H).

Preparation of 3- [2-N-BOC-piperidin-4-yl) ethyl] -3-acetyl-2-pyrrolidinone 1-acetic acid (38a)

Using the same procedure as in conversion 16 to 17, 38a (340 mg, 91%) was obtained from 37 (390 mg, 1.1 mmol) as a white foam.

TLC R _f = 0.39 (9: 0.5: 0.5 CH ₂ Cl ₂ / methanol / HOAc).

3- [2- (N-BOC-4-piperidin-4-yl) ethyl] -3-acetyl- (2-pyrrolidinone-1) acetyl- [3 (R) - [2- (indol-3-yl) ethyl]. ] Preparation of β-alanine ethyl ester (39a)

Using the same procedure as for conversion 15 to 16, 38a (310 mg, 0.92 mmol) was obtained as a yellow oil 39a (460 mg, 86%) after flash chromatography (silica gel, 85% ethyl acetate / hexane).

TLC R _f = 0.16 (85% ethyl acetate / hexane);

^L HB MR (300 MHz, CDCl ₃ ) δ 8.21 (pis, 1H), 7.58 (d, J = 8] Hz, 1H), 7.38 (d, J = 8) Hz, 1H), 7.20 (t, J = 8 Hz, 1H), 7.13 (t, J = 8 Hz, 1H), 7.05 (pis, 1H), 6.72 (d, J = 10 Hz, 1H), 4.35 (m, 1H) [Α], 4.13 (q, J = 7 Hz, 2H), 4.09 (m, 2H), 3.97 (d, J = 15) Hz, 1H), 3.86 (d, J = 15 Hz, 1H), 3.41 (m, 2H), 2.80 (m, 2H), 2.65 (m, 2H), 2.55 (dd, J = 1.5 Hz, 2H), 2 .45 (m, 1H), 2.24 (m, 1H), 2.00 am · -1.20 (m, 10H), 1.48 (s, 9H), 1.26 (t, J = 7) Hz, 3H), 1.04 (m, 1H).

3- [2- (N-BOC-4-piperidin-4-yl) ethyl] -3-acetyl- (2-pyrrolidinone-1) acetyl- [3 (R) - [2- (indol-3-yl) ethyl]. ] βalanine preparation (40a)

Using the same procedure as in the case of conversion to 16, 40a (360 mg, 86%) was obtained from 39a (460 mg, 0.79 mmol).

TLC R _f = 0.57 (9: 1: 1 CH ₂ Cl ₂ / methanol / HOAc);

Preparation of [3- [2- (piperidin-4-yl) ethyl-1] -acetyl- [3 (R) - [2- (indolyl) ethyl] β-alanine (41a)

Using procedure 18 of 17 from 40a (360 mg,

0.65 mmol) after flash chromatography (silica gel, 10: 0.4: 0.4 methanol / NH ₄ OH / water) gave 41a (180 mg, 59%) as a white solid.

TLC R _f = 0.21 (10: 0.4: 0.4 methanol / NH ₄ OH / water);

¹ H NMR (300 MHz, CD ₃ OD) 7.36 (d, J = 8 Hz, 1H), 7.13 (d,

J = 8 Hz, IH), 6.87 (t, J = 8 Hz, IH ), 6 .86 (s, 1H), 6.78 (t, J = 8 Hz, 1H), 4.09 (m, IH), 3.82 (d, J = 16 Hz, IH), 3.70 (d, J = 16 Hz, 1H), 3.33 (m, 2H), 3.07 (m, 2H), 2.63 (m, 4H), 2.30 (m, 1H), 2.25 (m, 2H), 2.05 (m, 1H), 1.90 -1.50 (m, 6H), 1.40-1.05 (m, 6H).

Preparation of [3- [2- (N-BOC-piperidin-4-yl) ethyl] -2-pyrrolidinone (36b)

Using procedure 36a to obtain from 35a, from 35b (0.7 g,

1.7 mmol) after flash chromatography (silica gel, 5% methanol / ethyl acetate) to give 36b (260 mg, 62%) as a solid white solid.

TLC = R _f = 0.27 (10% methanol / ethyl acetate).

Preparation of ethyl 3- [2-N-BOC-piperidin-4-yl) ethyl] -2-pyrrolidinone-acetate (37b)

Using procedure 37a from 36a, 36b (260 mg, 0.88 mmol) was obtained as an oil 37b (300 mg, 89%) after flash chromatography (silica gel, 50% ethyl acetate / hexane).

TLC R _f = 0.66 (10% methanol / ethyl acetate).

Preparation of 3- [2- (N-BOC-piperidin-4-yl) ethyl] -2-pyrrolidinone-1-acetic acid (38b)

Using procedure 17 of 17 out of 37b (300 mg,

0.78 mmol) was obtained as crystalline material 38b (260 mg, 98%).

TLC R _f = 0.38 (9: 0.5: 0.5 CH ₂ Cl / methanol / HOAc).

[3- [2- (N-BOC-piperidin-4-yl) ethyl] -2-pyrrolidinone-1] acetyl- [3 (R) - [2-pyrrolidinone-1] -acetyl- [3 (R) - Preparation of [2- (indol3-yl) ethyl] β-alanine ethyl ester (39b)

Using procedure 16 of 15 out of 38b (2 60 mg,

0.73 mmol) after flash chromatography (silica gel, 85% ethyl acetate / hexane) gave 39b as an oil (360 mg, 83%).

TLC R _f = 0.16 (85% methanol / ethyl acetate);

NMR (300 MHz, CDCl ₃ ) 8.29 (pis, 1H), 7.57 (d, J = 8 Hz, IH), 7.38 (d, J = 8 Hz, 1H) , 7.19 (t, J = 8 Hz, 1H), 7.12 (t, J = 8 Hz, 1H), 7.04 (d, J = 1 Hz, IH), 6.67 (d, J = 10 Hz, 1H), 4.33 (m, 1H) , 4.13 (sq, J = 7 Hz, 2H), 4.07 (m, 2H), 3.99 (d, J = 16) Hz, 1H), 3. 82 (d, J = 16 Hz, IH), 3.40 (m, 2H), 2.80 (m, 2H), 2.68 (m, 3H), 2.55 (dd, J = 5.2 Hz, 2H), 2. 43 (m, 1H), 2.22 (m, 1H), 2.00- -1.00 (m, 11H), 1.47 (s, 9H), 1.28 (t, J = 7Hz, 3H).

[3- [2- (N-BOC-piperidin-4-yl) ethyl] -2-pyrrolidinone-1] acetyl- [3 (R) - [2- (indol-3-yl) ethyl] β-alanine (40b) Receipt

Using procedure 17 to obtain 16 out of 39b (280 mg,

0.47 mmol) after flash chromatography (silica gel 3284 B gel, 9: 1: 1 CH ₂ Cl ₂ / methanol / HOAc) to give 40b as a white solid (250 mg, 94%).

TLC R _f = 0.21 (9: 1: 1 CH ₂ Cl ₂ / methanol / HOAc).

[3- [2- (N-piperidin-4-yl) ethyl] -2-pyrrolidinone-1] acetyl [3 (R) - [2- (indol-3-yl) ethyl] β-alanine (41b) ) receipt

Using procedure 18 of 17 out of 40b (250 mg,

0.44 mmol) after flash chromatography (silica gel, 10: 0.4: 0.4 methanol / NH ₄ OH / water) gave 40b (100 mg, 49%) as a solid white solid.

TLC R _f = 0.46 (10: 0.4: 0.4 CH ₂ Cl ₂ / methanol / HOAc);

¹ H NMR (300 MHz, CD ₃ OD) 7.25 (d, J = 8 Hz, 1H), 7.29 (d, J = 8 Hz, H), 7.03 (t, J = 8 Hz, 1H), 7 .02 (s, 1H), 6.95 (t, J = 8 Hz, 1H), 4.27 (m, 1H), 4.05 (d, J = 16 Hz, 1H), 3.81 (d, J = 16 Hz, 1H), 3.42 (m, 2H), 3.26 (m, 2H), 2.90-2.70 (m, 4H), 2.49 (m, 1H), 2.40 (d, J = 6 Hz, 1H), 2.22 (m, 1H), 2.05-1.70 (m, 5H), 1.60-1.25 (m, 5H). g) Swing 1-5idr.c Ū j ii Π r · In1 ί i b) ΡΗ, Γ OocN <_ / CCj et 9th 42

Ethyl 4- (N-BOC-piperidin-4-yl) crotonate (42)

DMSO (0.52 mL, 7.0 mmol) was added dropwise to a stirred solution of oxalyl chloride (0.43 mL, 5.0 mmol) in CH ₂ Cl ₂ (20 mL) at -78 ° C. When the gas ceases (~ 5 minutes), a stream of alcohol 9 (0.8 g, 3.5 mmol) is dissolved in CH ₂ Cl ₂ (20 mL). After 20 minutes, triethylamine (1.7 ml, 12 mmol) was added dropwise and the cooling bath was removed. After 20 minutes, carbethoxymethylenotriphenyl-phosphorane (1.4 g, 4.0 mmol) was added in one portion. After 2.0 hours, the reaction mixture was diluted with petroleum ether and then washed with H ₂ 0, 5% KHSO ₄ and brine, dried over MgSO ₄ , and concentrated.

Flash chromatography (silica, 15% ethyl acetate / hexane) affords the ester 42 as a colorless oil.

TLC R _f = 0.79 (50% ethyl acetate / hexane);

¹ H NMR (300 MHz, CDCl ₃ ) δ 6.91 (dt, J = 16 and 7 Hz, 1H), 5.81 (pld, J = 17 Hz, 1H), 4.18 (kv, J = 7 Hz, 2H), 4.08 (m, 2H), 2.67 (m, 2H), 2.14 (t, J = 7Hz, 2H), 1.071.05 (m, 5H), 1.44 (s, 9H), 1.28 (t, J = 7Hz, 3H).

Preparation of ethyl 4- (N-BOC-piperidin-4-yl) butyrate (43)

The olefin 42 (26 g, 87 mmol) in ethyl acetate (500 mL) was hydrogenated overnight at room temperature under a hydrogen atmosphere (1 atm) in the presence of 10% Pd / C (5.0 g). The reaction mixture was purged with argon and filtered through a pad of celite. Concentration of the filtrate gives the ester as a crystalline material by flash chromatography (silica, 10% ethyl acetate / hexane).

TLC R _f = 0.42 (20% ethyl acetate / hexane);

^X H NMR (300 MHz, CDC1 ₃ ) δ 4.16 (sq, J = 7 Hz, 2H), 4. 10 (m, 2H), 2. 69 (m, 2HO, 2.31 (t, J = 7 Hz, 2H), 1. 26 (m, 4H), 1.38 (s, 9H) , i. 40 (m, 1H), 1.11 (m, 2H).

Preparation of 4- (N-BOC-Piperin-4-yl) -butanoic acid (44)

A solution of ester 43 (19 g, 63 mmol), ethanol (300 mL) and 1 N NaOH (100 mL, 100 mmol) was stirred at room temperature for 2.5 h. The reaction mixture is then concentrated. The residue is diluted with 5% KHSO ₄ and ethyl acetate and transferred to a separatory funnel. The phases are mixed, separated and the organic phase is then washed with water, dried over MgSO ₄ and concentrated to give acid 44 as a colorless oil which crystallizes on standing.

mp = 80-81 ° C, TLC R _f = 0.68 (ethyl acetate);

¹ H NMR (300 MHz, CDCl ₃ ) δ 4.10 (m, 2H), 2.71 (m, 2H), 2.38 (t, J = 7Hz, 2H), 1.70 (m, 4H), 1.60-1.30 (m, 3H), 1.48 (s, 9H), 1.12 (m, 2H).

Preparation of 4 (S) -Benzyl-oxazolidinone-4- (BOC-piperidin-4-yl) butyrate (45)

To a solution of 44 (15.3 g, 56 mmol), NEt ₃ (9.4 mL, 67 mmol) and dry THF (240 ML) was added trimethylacetyl chloride (7.6 mL, 61 mmol) with stirring at -78 ° C. After 10 minutes. the cold bath is removed and replaced with an ice bath. After 1.0 hour, the heterogeneous mixture was again cooled to -78 ° C, then cooled to -78 ° C (S) - (-) - 4-benzyl-2-oxazolidone (62 mmol) in dry THF (150 mL) with n- BuLi (38.8 mL, 62 mmol, 1.6M / hexane). The addition of 1.0 hour, the reaction mixture temperature was increased to 0 ° C and the reaction mixture was diluted with ethyl acetate and washed with H ₂ O, dilute NaHCO _3, 5% KHSO ₄ and brine, dried over MgSO ₄ and concentrated. Flash chromatography (silica, 30% ethyl acetate / hexane) gives 45 as a colorless oil.

TLC R _f = 0.45 (30% ethyl acetate / hexane);

1 H NMR (400 MHz, CDCl ₃ ) 7.36-7.20 (m, 5H), 4.67 (m, 1H), 4.18 (m, 2H), 4.08 (m, 2H), 3.30 (dd, J = 13 and 3Hz, 1H), 2.93 (m, 2H), 2.77 (dd, J = 13 and 10 Hz, 1H), 2.69 (m, 2H), 1.70 (m, 4H), 1.50-1.30 (m, 3H), 1.45 (s) , 9H), 1.11 (m, 2H).

Preparation of 4 (S) -Benzyl-2-oxazolidone-4- (BOC-piperidin-4-yl) -2 (R) (2-cyanoethyl) butyrate (46)

To a solution of TiCl ₄ (42 mL, 42 mmol, 1M / CH ₂ Cl ₂ ) and CH ₂ Cl ₂ (250 mL) was added titanium (IV) isopropoxide (42 mL, 14 mmol) under stirring at 0 ° C. After 15 minutes add diisopropylethylamine (11.0 mL) dissolved in CH ₂ Cl ₂ (75 mL). The reaction mixture was stirred at 0 ° C until the solution turned dark red. After 10 minutes, 45 (21.8 g, 51 mmol) dissolved in CH ₂ Cl ₂ (75 mL) was added and stirred at 0 ° C for 1.0 h. At 0 ° C, add acrylonitrile (33.4 mL, 0.50 mol) to the dark red solution. After 4.0 hours, quench the reaction with dilute NH ₄ Cl (150 mL) at 0 ° C, then extract with CH ₂ Cl ₂ (3 x 250 mL). The combined organic extracts are washed with dilute NaHOC ₃ and brine, dried over MgSO _4, and concentrated. Flash chromatography (silica, 25% ethyl acetate / hexane) gives crude 46 as an oil. Purification by chromatography (silica, 2.5% acetone / CH ₂ Cl ₂ ) 46 combined with the mixed fractions (1.6 g) gave 46 as an oil, 97% pure by HPLC.

PSC R _f = 0.35 (2.5% acetone / CH ₂ Cl ₂ );

⁴ Η NMR (400 MHz, CDC1 ₃₎ δ 7:50 to 7:30 (m, 5H), 4.68 (m, IH), 4.21 (m, 2H), 4:07 (m, 2H), 3.80 (m, H),

3.33 (dd, J = 13 and 4 Hz, IH) , 2. 77 {dd, J = 13 & 10 Hz, IH), 2.65 (m, 2H), 2.38 (m, 2H), 2.13 (m, 1H), 1.89 (m, IH), 1.75 (m, IH), 1.63 (m, 2H), 1.50 (m, 2H), 1.45 (s, 9H), 1.35 (m, IH), 1.25 (m, 2H), 1.08 (m, 2H).

Preparation of 4 (S) -Cyclohexylmethyl-2-oxazolidin-one-4- (BOCpiperidin-4-yl) -2 (R) - (2-aminopropyl) butyrate-HCl (47)

77 Mixture, consisting of of 46 {19.2 g. 40 mmol), PtO ₂ (2.0g), CH ₃ OH (70 mL) and CHC1 ₃ {7.0 ml) , shake Parr in the apparatus hydrogen in the atmosphere (60 PSI) room

at a temperature of 3.0 hours. The reaction mixture was filtered through a celite filter and concentrated to give crude 47 amino-HCl as a white solid. This material is used directly in the next step.

TLC R _f = 0.50 (10: 1: 1 CH ₂ Cl ₂ / CH ₃ OH / HOAc);

⁷ Η NMR (400 MHz, CDCl ₃ ) δ 8.28 (bs, 2H), 4.50 (m, 1H),

4.35 (m, 1H), 4.14 (m, 1H), 4.03 (m, 2H), 3. 36 (m, 1H), 3.02 (m, 2H), 2 . 63 (m, 2H), 2am to 1pm (m, 24H), 1.45 (s, 9H).

Preparation of 3- (R) - [N-BOC-2- (piperidin-4-yl) ethyl] -2-piperidone (48)

Crude 47 amino.HCl (16.6 g, 31 mmol), acetonitrile (750 mL) and NaHCO ₃ (10.0 g) were stirred at room temperature for 20 hours. In order to block the small amount of piperidine formed in the previous reaction, the heterogeneous mixture is treated with di-tert-butyl dicarbonate (3.0 g) and stirred for a further 1.0 h. NaHCO _{3 is} removed by filtration and the filtrate is concentrated. Flash chromatography (quartz, 5% CH ₃ OH / ethyl acetate) affords the lactam in the form of 48 colorless crystals.

mp = 110-111 ° C; TLC R _f = 0.65 (20% CH ₃ OH / ethyl acetate;

⁷ H NMR (400 MHz, CDCl ₃ ) δ 6.31 (pis, 1H), 4.06 (m, 2H),

3.31 (m, 2H), 2.67 (m, 3H), 2.28 (m, 1H), 2.00-1.20 (m,

11H0, 1.45 (s, 9H), 1.10 (m, 2H).

Preparation of ethyl [3 (R) - [N-BOC-2- (piperidin-4-yl) ethyl] -2-piperidone-1] acetate (49)

To a stirred solution of 48 (6.7 g, 22 mmol) in dry THF (150 mL) was added dropwise NaN (TMS) ₂ (24.5 mL, 24.5 mmol, ΙΜ / hexane) at -78 ° C. After 15 minutes bromoacetate (5.2 mL, 45 mmol) was added and the reaction mixture was then brought to 0 ° C for 1 h. on the track. The reaction was then quenched with AcOH (1.0 mL), diluted with ethyl acetate, washed with H ₂ O and brine, dried over MgSO _4, and concentrated. Flash chromatography (silica, 40% ethyl acetate / hexane) affords the ester as a yellow oil 49.

TLC R _f = 0.26 (40% ethyl acetate / hexane);

^{Σ Η} NMR (300 MHz, CDC1 ₃₎ δ 4.20 (q, J = 7 Hz, 2H), 4.17 (d, J = 18 Hz, 1H), 4:08 (m, 2H), 3.98 (d, J = 18 Hz , 1H), 3.37 (m, 2H), 2.68 (m, 2H), 2.32 (m, 1H), 2.001.25 (m, 11H), 1.45 (s, 9H), 1.30 (t, J = 7Hz), 3H), 1.11 (m, 2H).

Preparation of 3- (R) - [N-BOC-2- (Piperidin-4-yl) ethyl] -2-piperidone-1] acetic acid (50)

A solution of 49 (6.0 g, 15 mmol), IN NaOH (50 mL, 50 mmol), and CH ₃ OH (75 mL) was stirred for 1 h. at room temperature. The reaction mixture was acidified with 5% aqueous KHSO ₄ and extracted with ethyl acetate. The organic portion was washed with brine, dried over MgSO _4, and concentrated to give the carboxylic acid 50 as a yellow oil. Addition of the oil produces a friable white material.

TLC R _f = 0.31 (9: 0.5: 0.5 CH ₂ Cl ₂ / CH ₃ OH / AcOH);

¹ H NMR (400 MHz, CDCl ₃ ) δ 4.13 (d, J = 17 Hz, 1H), 4.07 (m, 2H), 4.06 (d, J = 17 Hz, 1H), 3.39 (m, 1H), 2.33 (m, 1H), 1.95 (m, 3H), 1.81 (m, 1H), 1.70-1.25 (m, 7H), 1.45 (s, 9H), 1.08 (m, 2H).

Preparation of 3- (R) -BOC-amino-1-azobutan-2-one (52)

To a stirred solution of 51 (3.8 g, 20 mmol)

To 4-methylmorpholine (2.2 mL, 20 mmol) and ethyl acetate (200 mL) was added isobutyl chloroformate (2.6 mL, 20 mmol) at -15 ° C. After 1.0 or. the reaction mixture was washed with H ₂ O and brine, dried over MgSO ₄ and filtered into a round-bottomed flask. After cooling to 0 ° C, anhydrous diazomethane (80 mL, 40 mmol, 0.5 M solution) was added portionwise to the mixed anhydride. After 2 or. removable cooling bath and remove excess diazomethane by purging the solution for 30 min. running argon. Concentration gives crude diazoketone 52 which is used directly in the next step.

TLC R _f = 0.37 (30% ethyl acetate / hexane)

Preparation of ethyl N-BOC-3 (R) -Methyl β-alanine (53)

The crude diazoketone 52 (4.3 g, ~ 20.0 mmol) was dissolved in ethanol (250 mL) and NEt ₃ (3.4 mL, 24 mmol) and silver benzoate (1.4 g, 6.0 mmol) were added, resulting in vigorous gas evolution and black precipitation. After 1 or. the reaction mixture is concentrated and purified by flash chromatography (silica, 10% ethyl acetate / hexane) to give the ethyl ester 52 as a colorless oil.

TLC R _f = 0.42 (30% ethyl acetate / hexane);

^{Χ Η} NMR (400 MHz, CDC1 ₃₎ 5 4.96 (m, 1H), 4.19 (q, J = 7 Hz, 2H), 4:04 (m, 1H), 2:52 (dd, J = 15 and 6 Hz, 1H) , 2.46 (dd, J = 15 and 6 Hz, 1H), 1.44 (s, 9H), 1.26 (t,

J = 7Hz, 3H), 1.21 (t, J = 7Hz, 3H).

Preparation of ethyl-3 (R) -Methyl-3-alanine-HCl (54)

To a mechanically stirred solution of 53 (2.2 mg, 9.7 mmol) in ethyl acetate (180 mL) at -15 ° C for 30 min. gaseous HCl is actively allowed. Then remove the cooling bath and the solution for 1 hour. purged with argon to remove excess HCl. Concentration gave the amine hydrochloride 54 as a yellow oil.

1 H NMR (400 MHz, D ₂ O) δ 4.21 (q, H = 7 Hz, 2H), 3.78 (m,

1H), 2.79 (m, 2H), 1.38 (d, J = 7Hz, 1.29 (t, J = Hz, 3H).

(R) - [(N-BOC-2- (piperidin-4-yl) ethyl] -2-piperidinone] acetyl- [3 (R) -methyl-p-alanine ethyl ester (55a) (1.0g, 2.7) mmol), (0.39 g, 2.8 mmol), and dry DMF (100 mL) was added to EDC (0.95, After 20 h acetate and aqueous

HOBT was added to a solution of 54 (0.48 g, 2.8 mmol).

N (i-pr) ₂ Et (1.5 mL, 8.5 mmol)

At -15 ° C with stirring

2.8 mmol) followed by removal of the cooling bath, the reaction mixture was diluted with ethyl acetate, washed with water, saturated with NaHCO ₃ , 5%

KHSO ₄ , brine, dried over MgSO ₄ and concentrated. Flash chromatography (silica, ethyl acetate) gave 55 as a colorless oil.

TLC R _f = 0.35 (ethyl acetate);

^{Σ Η} NMR (300 MHz, CDC1 ₃₎ δ 6.82 (bd, 1H), 4:32 (m, 1H),

4.13 (s, J = 7 Hz, 2H), 4.10 (d, J = 15 Hz, 1H), 4.08 (m,

2H), 4.82 (q, J = 15 Hz, 1H), 3.36 (m, 2H), 2.67 (m, 2H),

2.48 (dd, J = 5 and 1 Hz, 2H), 2.33 (m, 1H), 2.00-1.20 (m, 11H), 1.45 (s, 9H), 1.27 (t, J = 7 Hz, 3H), 1.21 (d, J = 7Hz, 3H), 1.10 (m, 2H).

Preparation of (R) - [(N-BOC-2- (piperidin-4-yl) ethyl] -2-piperidinone] acetyl- [3 (R) -methyl-p-alanine (56)]

A solution of 55 (1.2 g, 2.5 mmol), IN NaOH (10 mL, 10 mmol) and CH ₃ OH (18 mL) was stirred for 1 h. at room temperature. The reaction mixture was acidified with 5% aqueous KHSO ₄ and extracted with ethyl acetate. The organic layer was washed with brine, dried over MgSO _4, and concentrated. Purification by flash chromatography (silica gel, 10: 0.5: 0.5 CH ₂ Cl ₂ / CH ₃ OH / AcOH) afforded aceotropic distillation of the AcOH with toluene to afford the carboxylic acid 56 as a colorless oil.

TLC R _f = 0.40 (10: 0.5: 0.5 CH ₂ Cl ₂ / CH ₃ OH / AcOH);

¹ H NMR (300 MHz, CDCl ₃ ) δ 7.02 (bd, 1H), 4.35 (m, 1H),

4.12 (d, J = 7Hz, 1H), 4.08 (m, 2H), 3.87 (d, J = 16Hz, 1H), 3.32 (m, 2H), 2.69 (m, 2H), 2.56 (m, 2H) ), 2.36 (m, 1H), 2.00-1.25 (m, 11H), 1.45 (s, 9H), 1.03 (m, 2H).

Preparation of (R) - [2- (piperidin-4-yl) ethyl] -2-piperidone-1] acetyl [3 (R) -methyl-p-alanine (57)]

A solution of 57 (0.74 g, 1.6 mmol), trifluoroacetic acid (10 mL) and CH ₂ Cl ₂ (10 mL) was stirred for 1 h. at room temperature. The reaction mixture is concentrated and the remaining trifluoroacetic acid is removed by aceotropic distillation with toluene. Flash chromatography (silica gel, 10: 0.5: 0.5 CH ₃ OH / NH ₄ OH / H ₂ O) gives 57 as a white amorphous substance. Recrystallization of 57 (100 mg) from ethanol (4 mL) and filtration at room temperature gives (45 mg) in crystalline form.

mp = 240 ° (s) PSC R _f = 0.48 (10: 1: 1 CH ₃ OH / NH ₄ OH / H ₂ O);

¹ H NMR (400 Hz, D ₂ O) δ 4.18 (m, 1H), 4.06 (d, J = 7 Hz,

1H), 3.9J 5 (d, J = 16 Hz, 1H), 3. 41 (m, 4H), 2.99 (m, 2H), 2.45 (m, 1H), 2.42 (dd, J = 18 and 6 Hz, 1H), 2.33 (dd, J = 18 and 7 Hz, 1H), 1.97 (m, 4H), 1.90-1.55 (m, 5H), 1.38 (m, 4H), 1.19 (d, J = 7 Hz, 3H).

- Preparation of [2- (N-BOC-piperidin-4-yl) ethyl] -3-propen-2-yl- (1H-tetrahydropyrimidin-2-one) (59)

To a stirred solution of 1H-tetrahydropyrimidin-2-one (1.0 g, 10 mmol) in DMF (50 mL) was added a 1 M solution of lithium bis (trimethylsilyl) amide (10 mL, 10 mM). After 0.5 or. allyl bromide (1.0 mL, 1.17 mM) was added and stirring was continued for 1 h. A second portion of lithium bis (trimethylsilyl) amide (10 mL, 10 mM) was then added, followed by 0.5 h. (N-BOC-piperidin-4-yl) ethyl iodide (10) is added. The reaction mixture was stirred at 25 ° C overnight. The reaction mixture was poured onto ice and 15% KHSO ₄ (50 mL), extracted with ethyl ether (2 x 100 mL), washed with H ₂ O ( ₂ x 100 mL), brine, dried over Na ₂ SO ₄ and evaporated in vacuo. Flash chromatography on SiO ₂ (ethyl acetate-hexane 1: 1) gives pure 59.

¹ H NMR (300 MHz, CDCl ₃ ) δ 1.14 (2H, m), 1.46 (9H, s), 1.74 (3H, m), 1.95 (2H, m), 2.68 (2H, t), 3.22 (4H, m), 3.38 (2H, m), 3.95 (2H, m), 4.06 (2H, m), 5.13 (2H, dd).

Preparation of 1- [2- (N-BOC-piperidin-4-yl) ethyl] -3-acetaldehyde- (1 H-tetrahydropyrimidin-2-one) (60) A solution of THF (7 mL), H ₂ O (5 mL), periodate (0.68 g, 3.19 mM), a solution of 2-methyl-2? 59 (430 mg, 1.27 mM) in sodium 2.5% osmium (4.13 µl, tetroxide propanol (4.13 µl, 1.28 mM)) was stirred for 4 hours, ethyl acetate was added. (70 ml) and washed with 50% sodium chloride / 10% sodium sulfate (1: 1) and brine, dried over Na ₂ SO ₄ to give 60.

The reaction mixture is evaporated in vacuo to give 1- [2- (N-BOC-piperidin-4-yl) ethyl] -3-acetic acid- (1 H-tetrahydropyrimidin-2-one) (61)

To a solution of 60 (374 mg) in acetone (7 mL) cooled to -15 ° C was added Jonah's reagent until the orange color persisted for 5 min. Ethyl acetate (60 mL) was added, the solution was washed with H ₂ O, brine, dried over NaSO ₄ and evaporated in vacuo to give 61.

¹ H NMR (300 MHz, CDCl ₃ ) δ 1.12 (2H, m), 1.45 (9H, s),

1.70 (2H, pld), 2.03 (2H, m), 2.70 (2H, plt), 3.37 (4H, m), 3.95 (2H, m), 4.05 (2H, m).

Preparation of 1- [2- (N-BOC-piperidin-4-yl) ethyl] - (1 H-tetrahydropyrimidin-2-one) -3-acetyl-3 (R) - (2-phenylethyl) β-alanine methyl ester (63)

A mixture of 61 (125 mg, 0.34 mM), (R) -3 (2-phenylethyl) β-alanine methyl ester (83 mg, 0.34 mM),

1-Hydroxybenzotriazole hydrate (48.3 mg, 0.35 mM), 1-ethyl-3- (3-dimethylaminopropyl) -carbodiimide hydrochloride (70 mg, 0.37 mM) and triethylamine (107 µL, 0.77 mM) were stirred in dry DMF (1.6 mL) for 18 h. At 25 ° C. The reaction mixture was poured onto ice, extracted with ethyl ester (2x50 mL), washed with 10% citric acid, brine, dried over Na ₂ SO ₄ and evaporated in vacuo to give 63.

1 H NMR (300 MHz, CDCl ₃ ) δ 1.14 (2H, m), 1.46 (9H, s), 1.7 (2H, m), 1.86 (2H, m), 2.02 (2H, m), 2.58 (2H, d), 2.66 (3H, m), 3.36 (5H, m), 3.68 (2H, m), 3.93 (IH, d), 4.07 (2H, m), 7.02 (1H, m), 7.20 (2H, m), 7.29 (2H, m).

- Preparation of [2- (N-BOC-piperidin-4-yl) ethyl] - (1 H-tetrahydropyrimidin-2-one) -3-acetyl-3 (R) - (2-phenylethyl) β-alanine (64) (168 mg, 0.31) mM) solution in water (15 mL) was cooled to 0 ° C, IN LiOH (1.2 mL) was added and stirred overnight at 25 ° C. THF is then evaporated in vacuo, the residue is acidified with 10% citric acid, extracted with ethyl acetate (2x50 mL), washed with brine, dried over Na ₂ SO ₄ and evaporated in vacuo to 64.

Preparation of 1- [2- (N-BOC-piperidin-4-yl) ethyl] - [(1 H-tetrahydropyrimidin-2-one) -3] -acetyl-3 (R) - (2-phenylethyl) β-alanine (65) mg) solution of methylene chloride (8 ml) and anisole (1.5 ml) was cooled to -20 ° C for 5 min. Trifluoroacetic acid (8 ml) was added over the course. The reaction mixture was stirred for 1 h. At -5 ° C, then evaporated in vacuo. The residue is chromatographed on SiO ₂ (ethanolH ₂ O NH ₂ OH) to give 65. Analysis calculated for C ₂₄ H ₃₆ N ₄ O ₄ * 2.5H ₂ O:

C, 57.84; H, 8.65; N, 11.73

Found: C, 57.94; H, 8.39, N, 11.47 1- [2- (N-BOC-piperidin-4-yl) ethyl] - [(1H-tetrahydro-pyrimidin-2-one) -3] -acetyl-3 (R) - (2-indole- 3il) -ethyl] β-alanine (66)

Basically according to the procedure described for Preparation 63, except that an equivalent amount of 3 (R) - (2-indol-3-ethyl) β-alanine ethyl was used instead of 3 (R) - (2-phenylethyl) β-alanine methyl ester hydrochloride. of the ester and 66 according to the procedures described for 64 and 65.

Analysis calculated for C ₂₆ H ₃₇ N ₅ O ₄ :

C, 56.3; H, 8.0, N, 12.6

Found: C, 56.3; H, 7.9, N, 12.2 1- [2- (N-BOC-piperidin-4-yl) ethyl] - [(1H-tetrahydropyrimidin-2-one) -3] -acetyl-3 (R) -methyl-β-alanine (66 ) receipt

Basically according to the procedure described for the preparation of 63, except that the equivalent amount of 3 (R) -methyl-4-alanine methyl ester hydrochloride was used instead of 3 (R) - (2-phenylethyl) β-alanine methyl ester hydrochloride and according to the procedures described in 64. and for 65, 67 were obtained.

1- [2- (N-BOC-piperidin-4-yl) ethyl] - [(1H-tetrahydropyrimidin-2-one) -3] -acetyl-2 (R) - [butylsulfonylamino] -β-alanine (66) receipt

Basically according to the procedure described for the preparation of 63, except that the equivalent amount of 2 (S) -butylsulfonylamino-4-alanine hydrochloride was used instead of 3 (R) - (2-phenylethyl) β-alanine methyl ester hydrochloride and according to the procedures described in 64. and 65 for receipt, 68 was obtained.

Preparation of methyl 4 (R) -benzyl-2-oxazolidinone succinate (70)

Trimethylacetyl chloride (3.6 mL, 28.2 mmol) was added dropwise to a solution of mono-methyl succinate 69 (3.7 g, 28.2 mmol), NEt ₃ (4.0 mL, 28.2 mmol) and THF (160 mL) at -78 ° C. After 10 minutes. removable refrigerating bath and replace with ice bath. After 1 or. the heterogeneous mixture was cooled again to -78 ° C and a solution of lithium (R) - (+) - 4-benzyl-2-oxazolidinone (28.2 mmol) in dry THF (43 mL) obtained by treatment with (R) - (+) - 4-Benzyl-2-oxazolidinone (5.0 g, 28.2 mmol) in dry THF (43 mL) at -78 ° C was treated with n-BuLi (17.8 mL, 28.2 mmol).

1.6 M / Hexane). After addition, the reaction mixture is warmed to room temperature overnight. The reaction mixture was then diluted with ethyl acetate and washed with H ₂ O, saturated NaHCO ₃ , 5% KHSO ₄ and brine, dried over MgSO ₄ and evaporated in vacuo. Flash chromatography (silica, ethyl acetate-hexane 1: 1) gives pure 70 as a white crystalline solid.

TLC R _f = 0.48 (30% ethyl acetate / hexane);

¹ H NMR (300 MHz, CDCl ₃ ) δ 7.30 (m, 2H), 4.68 (m, 1H), 4.21 (m, 2H), 3.72 (s, 3H), 3.28 (m, 2H), 2.80 (dd, J = 13 Hz and 4 Hz, 1H), 2.73 (m, 2H)

Preparation of methyl 4 (R) -benzyl-2-oxazolidinone-2 (R) - (2-cyanoethyl) succinate (71) in TiCl ₄ (12.3 mL, 12.3 mmol, 1M CH ₂ Cl ₂ ) CH ₂ Cl ₂ (83 of isopropoxide (1.2 mL, 4.1 mmol) after oral administration of diisopropylethylamine (3 mL, dark brown solution) at 70 ° C (4.8 mL, 16.5 mmol) and then at room temperature in dark Ti (IV) after 15 min. (juice 17.3 mmol) is obtained for 10 min, a solution of acrylonitrile (4.3 ml, 66 mmol) is added dropwise with stirring at 0 ° C and stirring is continued for 72 hours. At 0 ° C. The reaction is quenched by addition of a saturated solution of NH ₄ Cl at 0 ° C and extracted with CH ₂ Cl ₂ . The combined organic layers were washed with saturated NaHCO ₃ , brine, dried over MgSO _4, and concentrated. Flash chromatography (silica, 25% ethyl acetate / hexane) gave 71 crystals.

TLC R _f = 0.22 (20% ethyl acetate / hexane);

^{Χ Η} NMR (300 MHz, CDC1 ₃₎ H), 4.25 (m, 2H);

3.30 (dd, J = 13 Hz and 10 Hz, 1H), 2.80 (dd,

J = 13 and 4 Hz, 1H), 2.43

1.90 (m, 1H).

δ 7.40-7.20 (m, 5H), 4.70 (m,

4.13 (m, 1H), 3.67 (s, 3H),

Hz, 1H), 2.93 (dd, J = 17 and

J = 17 and 10 Hz, 1H), 2.55 (dd, (t, J = 7 Hz, 2H), 2.10 (m, 1H),

Preparation of methyl 4 (R) -benzyl-2-oxazolidinone-2 (R) - (3-aminoprop: succinate) (72)

A mixture of 71 (2.0 g, 5.8 mmol), PtO ₂ (0.8 g), CH ₃ OH (50 mL), and CHCl ₃ (5 mL) is shaken in a Parr apparatus at room temperature under hydrogen (60 PSI) for 3 hours. The reaction mixture was filtered through a pad of celite and concentrated to give crude 72 amino * HCl as a yellow oil.

TLC R _f = 0.50 (CH ₂ Cl ₂ / CH ₃ OH / AcOH 9: 1: 1)

Preparation of methyl [2-Piperidone-3 (R)] acetate (73)

Crude 72 amino * HCl (2.0 g, ~ 5.8 mmol), acetonitrile (250 mL) and NaHCO ₃ (5 g) were stirred overnight at room temperature. The reaction mixture is then filtered and concentrated. Flash chromatography (silica gel, ethyl acetate) affords the lactam in the form of 73 colorless crystals.

TLC R _f = 0.22 (ethyl acetate);

^{Χ Η} NMR (300 MHz, CDCl ₃₎ 5 5.82 (bs, IH), 3.70 (s, 3H);

3.33 (m, 2H), 2.88 (dd, J = 16 and 4 Hz, 1H), 2.75 (m, 1H), 2.58 (dd, J = 16 Hz and 7 Hz, 1H), 2.10-1.60 (m, 4H) ).

Preparation of methyl [1-N-BOC-2- (piperidin-4-yl) -ethyl] -2-piperidone-3 (R)] acetate (74)

To a stirred solution of 73 (1.1 g, 6.4 mmol) in anhydrous DMF (60 mL) was added dropwise NaN (TMS) ₂ (6.4 mL, 6.4 mmol, ΙΜ / hexane) at -15 ° C, then the cooling bath was removed. After 10 minutes, iodide (2.3 g, 7.0 mmol) was added and stirred at room temperature for 4 hours. The reaction mixture was then diluted with ethyl acetate and washed with H ₂ O (2x) and brine, dried over MgSO _4, and concentrated. Flash chromatography (silica gel, 40% ethyl acetate / hexane) gave 74 as an oil.

TLC R _f = 0.29 (40% ethyl acetate / hexane).

Preparation of [1-N-BOC-2- (Piperidin-4-yl) -ethyl] -2-piperidone-3 (R)] -acetic acid (75)

Using the procedure of obtaining 50 from 49, 75 (0.44 g, 1.1 mmol) are obtained in the form of 75 oils.

TLC R _f = 0.52 (CH ₂ Cl ₂ / CH ₃ OH / AcOH 9: 0.5: 0.5).

Preparation of [1- [N-Boc-2- (Piperidin-4-yl) -ethyl] -2-piperidone-3 (R)] -acetyl-3 (R) -methyl-3-alanine ethyl ester (76).

Using the procedure of Preparation 55 from 50, 75 (0.40 g, 1.1 mmol) was coupled to 54 (0.27 g, 1.6 mmol) and obtained after flash chromatography (silica gel, ethyl acetate) (76) as an oil.

TLC R _f = 0.20 (ethyl acetate).

Preparation of 1- [N- (Boc-2- (piperidin-4-yl) ethyl] -2-piperidone-3 (R)] acetyl-3 (R) -ethyl-β-alanine (77)

Using Procedure 56 to obtain 55 from 55, 76 (0.30 g, 0.64 mmol) are obtained in the form of 77 as an oil.

TLC R _f = 00:24 (CH ₂ C1 ₂ / CH ₃ OH / HOAc 9: 0.5: 0.5), ^X H NMR (400 NHZ, CDC1 ₃₎ δ 6.93 (m, H), 4:32 (m, H),

4.06 (m, 2H), 3.35 (m, 2H), 3.29 (m, 2H), 2.70-2.50 (m, 6H), 2.38 (m, 1H), 2.00-1.20 (m, 9H), 1.45 (s, 9H),

1.27 (d, J = 6Hz, 3H), 1.06 (m, 2H).

Preparation of [1- [2- (piperidin-4-yl) ethyl] -2-piperidone-3 (R) -methyl-β-alanine (78)

Using the procedure of Preparation 57 from 56, 77 (0.24 g, 0.53 mmol) were obtained as amorphous material after flash chromatography (silica, 10: 1: 1 / ethanol / H ₂ O / NH ₄ 0 H).

^X H BM R (400 MHz, D ₂ O) δ 4. 19 (m, 1H), 3.40 (m, 6H) 3.00 (m, 2H), 2.70 (m, IH), 2.64 (m, 1H), 2.47 (m, 2H) 2.30 (dd, J = 14 and 8Hz, IH), 2.00 (m, 2H), 1.92 (m, 2H) 1.79 (m, 1H), 1. 61 (m.) r 4H) 1.45 (m, 2H), 1.18 (d J = 7Hz , 3H).

Preparation of N-Cbz-Gly-3 (R) -methyl-p-alanine ethyl ester (79)

To a solution of Cbz-Gly (0.94 g, 4.5 mmol), 4-methylmorpholine (1.1 mL, 4.9 mmol) and ethyl acetate (50 mL) was added isobutyl chloroformate (0.61 mL, 4.7 mmol) at -15 ° C. (add up immediately After 15 min add 54 (0.75g, 4.5 mmol) in total). solution was washed with ethyl room acetate and NaHCO ₃ at a temperature of 1M NaHSO ₄ (1 ml) overnight then the reaction salts ^ - solution, brine, dried mix is considered dilute

MgSO ₄ , and upon concentration gives 79 as a yellow oil.

^{Χ Η} NMR (300 MHz, CDCL ₃₎ δ 7:48 (m, 5H), 6.73 (m, 1H), 5:58 (m, 1H), 5.13 (s, 2H) and 4.48 (m, 1H), 4.30 (q, J = 7Hz, 2H), 2.62 (m, 2H), 1.37 (t, J = 7Hz, 3H).

Preparation of the ethyl salt of Gly-3 (R) -methyl-3-alanine (80) with acetic acid ester

A mixture of 79 (1.4 g, 435 mmol), 10% AcOH / CH ₃ OH (50 mL) and 20% Pd (OH) ₂ was shaken overnight on a Parr apparatus under hydrogen atmosphere (60 PSI). After 24 or. the reaction mixture is filtered through a pad of celite and the resulting filtrate is concentrated. Flash chromatography (silica, 5: 3: 1 CH ₂ Cl ₂ / ethanol / HOAc) gives 80 as an amorphous material.

Preparation of (S) -Benzyl-2-oxazolidone-4- (N-BOC-piperidin-4-yl) 2 (R) - (prop-2-en) butyrate (81)

To a stirred solution of 45 (2.5 g, 5.8 mmol) in THF (50 mL) at -78 ° C was added lithium bis (trimethylsilyl) amino (7.0 mL, 7.0 mmol, ΙΜ / hexane), followed by addition of allyl bromide ( 2.5 mL, 29 mmol). After adding the above materials, remove the cooling bath and stir at 0 ° C for 1.5 hours. The reaction was quenched with dilute NH ₄ Cl, diluted with EtOAc, washed with dilute NaHCO ₃ , 5% KHSO _4, and brine, dried over MgSO _4, and concentrated. Flash chromatography (silica, 15% EtOAc / hexane) gave 81 as a colorless oil.

PSC R _f = 0.53 (30% ETOAc / hexane);

¹ H NMR (300 NHz, CDCL ₃ ) δ 7.40- 7.20 (m, 5H), 5.82 (m, IH), 5.10 (m, 2H), 4.80 (m, 1H), 4.20 (m, 2H), 4.07 (m, 2H), 3.90 (m, 1H), 3.31 (dd, J = 13 and 3 Hz, IH), 2.68 (m, 2H), 2.48 (m, IH), 2.35 (m, 1H), 1.90 -1.20 (m, 7H), 1.47 (s, 9H), 1.10 (m, 2H).

Preparation of 4- (N-Bocpiperidin-4-yl) -2 (R) - (prop-2-en) butyric acid (82)

To a solution of 81 (1.8 g, 3.8 mmol), 30% H ₂ O ₂ (8.5 mL, 83 mmol), THF (41 mL) and H ₂ O (12 mL) was added LiOH (14 mL, mmo1, zuoj amas solution.KHSO ₄ and

IN). After 2 hours at 0 ° C, the reaction was extracted

Excess LiOH at neutral temperature Acidify 10% NaHSO _{4 with} 5% EtOAc. The organic layer of EtOAc was washed with brine, dried over MgSO _4, and concentrated to give 82 as a colorless oil.

TLC R _f = 0.79 (10% CH ₃ OH / EtOAc).

Preparation of 4- (N-BOC-piperidin-4-yl) -2 (R) - (prop-2-ene) butyrate] Gly-3 (R) -methyl-p-alanine ethyl ester (83)

Using a procedure analogous to 50 for conversion to 55, 82 (1.3 g, 2.5 mmol) was coupled to 80 (0.62 g, 2.5 mmol) and 83 after flash chromatography (silica, 10% isopropanol / hexane).

TLC R _f = 00:50 (20% isopropanol / hexanes);

^Χ Η B MR (300 6.37 (m, IH) 4.13 (q, J = 7 2.63 (m, 2H), 2.20 (m, 2H), J = 15 Hz, 3H),

NHz, CD Cl ₃ ) 5.72 (m, IH), Hz, 2H) , 4 2.51 (m, 2H) 1.70 -1.2 0 (m 1.07 (m, 2H)

δ 6.67 (bd, 5.02 (m, 1H), 03 (m, 1H), 2.51 (m, 2H), 7H), 1.44 (s,

J = 8 Hz, IH), 4.35 (m, IH), 1.88 (m, 2H), 2.35 (m, IH), 9H), 1.26 i (t,

Preparation of [3- [2- (N- (BOC-Piperidin-4-yl) ethyl] -50-hydroxy-2-pyrrolidinonone-1] acetyl-piperidone-3 (R) -methyl-p-alanine ethyl ester (84)

To a solution of 83 (0.74 g, 1.5 mmol), THF (12.2 mL), H ₂ O (9.2 mL) and NaIO ₄ (0.82 g, 3.8 mmol) was added OsO ₄ (0.96 mL, 80 µmol) at room temperature; % tert-butanol). After 2 or. the reaction mixture is diluted with ether and washed with H ₂ 0, 10%

Na ₂ S ₂ O ₃ , dried with brine (MgSO ₄ ) and concentrated to give 84 as a yellow oil. TLC R _f = 0.26 (20% isopropanol / hexane) [3- [2- (N- (BOC-Piperidin-4-yl) ethyl] -5-hydroxy-2-pyrodinonone-1] acetylpiperidone-3 (R) -methyl-). yielding p-alanine (85)

Using the same procedure as in 56 out of 55, out of 84 (0.73 g, 1./5 mmol) 85 were obtained in the form of pale yellow foam.

1 H NMR (300 MHz, CDCl ₃ ) δ 5.23 J = 16 Hz, 1H) 4.32 (m, IH), J = 16 Hz, 1H), 2.80-2.40 (m, 1.00 (m, 9H), 1.45 (s, 9H), 1

(d, J = 5Hz, 1H), 4.58 (d, 4.05 (m, 2H), 3.70 (d,

6H), 2.22 (m, 1H), 2.0027 (t, J = 15 Hz, 3H) [3- [2- (N- (Piperidin-4-yl) ethyl] -2-pyrrolidinonone1] acetyl-3 (R) Preparation of -methyl-3 ~ -alanine (86)

To a stirred solution of 85 (0.61 g, 1.3 mmol) in CH ₂ Cl _{2 was} added CF ₃ COOH (5.8 mL) and triethylsilane (0.8 mL, 5.2 mmol) at room temperature. After 2 or. the reaction mixture is concentrated and the remaining CF ₃ COOH is removed by aceotropic distillation with toluene. Flash chromatography (silica, 10: 1: 1 ethanol / H ₂ O / NH ₄ 0H) gives 86 as a colorless solid.

TLC R _f = 0.17 (9: 1: 1 ethanol / H ₂ O / NH ₄ O H);

⁴ Η NMR (300 MHz, CD ₃ OD) δ 4.20 (m, 1H), 3.97 (d, J = 17 Hz, 1H), 3.85 (d, J = 17 Hz, 1H), 3.40 (m, 4H);

2.95 (m, 2H), 2.52 (m, 1H), 2.30 (m, 2H), 2.00-1.30 (m, 11H), 1.20 (d, J = 7Hz, 3H).

Preparation of 2- (2,3,4,5,6-tetrahydro-1,1-dioxo-2H-1,2-thiazin-2-yl) prop-2-ene (88)

To a solution of 87 (JOC 52, p. 2162 (1987) EHWhite, HMLim) (0.35 g, 2.6 mmol) in DMF at 0 ° C was added NaN (TMS) ₂ (3.9 mL, 3.9 mmol); 1M solution in hexane. After 30 min, add allyl bromide (0.45 mL, 5.2 mmol) and stir for 30 min. The reaction mixture was diluted with ethyl acetate, washed with saturated NH ₄ Cl and brine, dried over MgSO _4, and concentrated. Flash chromatography (silica, 30% ethyl acetate / hexane) gave 88 as a colorless oil.

TLC R _f = 0.29 (30% ethyl acetate / hexane) ¹ H NMR (300 MHz, CDCl ₃ ) δ 5.80 (m, 1H), 5.25 (m, 2H),

3.78 (m, 2H), 3.30 (m, 2H), 3.02 (m, 2H), 2.21 (m, 2H),

1.68 (m, 2H).

2- [6- [2- (N-BOC-Piperidin-4-yl) ethyl] - (3,4,5,6-tetrahydro-1,1-dioxo-2H-1,2-thiazin-2-yl) prop Obtaining -2-ene (89)

To a stirred solution of 88 (0.34 g, 1.93 mmol) in THF (10 mL) at -78 ° C was added dropwise n-BuLi (1.45 mL,

2.3 mmol; 1.6 M / Hexane) solution. After 30 minutes to the reaction mixture sequentially add l-methyl-2-pyrrolidone (1 mL) and 10 (0.86 mL, 2.7 mmol) THF (10 mL) and raise the temperature to -23 ° C. The reaction mixture is kept for 2 hours. At -23 ° C and quenched with saturated NH ₄ Cl solution. The reaction mixture was then diluted with ethyl acetate and washed with H ₂ O and brine, dried and concentrated. Flash chromatography (silica, 30% ethyl acetate / hexane) gave 89 crystals.

TLC R _f = 0.27 (30% ethyl acetate / hexane);

¹ H NMR (400 MHz, CDCl ₃ ) δ 5.80 (m, 1H), 5.25 (m, 2H),

4.07 (m, 2H), 3.78 (m, 2H), 3.34 (m, 2H), 3.17 (m, 1H),

3.02 (m, 1H), 2.82 (m, 2H), 2.66 (m, 2H), 2.24-2.04 (m,

3H), 1.89 (m, 2H), 1.70-1.35 (m, 7H), 1.08 (m, 2H).

2- [6- [2- (N-BOC-Piperidin-4-yl) ethyl] - (3,4,5,6-tetrahydro-1,1-dioxo-2H-1,2-thiazin-2-yl) acetaldehyde (90) Preparation of a solution of 89 (0.50 g, 1.29 mmol), THF (16 mL), H ₂ O (12 mL) and NaIO ₄ (0.69 g, 3.2 mmol) was added OSO ₄ (1.0 mL, 0.1 mmol; 2.5% tertbutanol). After 3 or. the reaction mixture was diluted with ethyl acetate and washed with H ₂ O, 5% KHSO ₄ , saturated NaHCO ₃ , brine, dried over MgSO _4, and concentrated to give the aldehyde 90 as an oil.

95 ¹ H NMR (400 MHz, CDCl ₃ ) δ 9.64 (s, 9H), 4.07 (m, 2H), 4.00 (d, J = 17 Hz, 1H), 3.89 (d, J = 17 Hz, 1H), 3.62 (m, 1H), 3.14 (m, 1H), 2.96 (m, 1H) , 2.66 (m, 2H), 2.20 (m, 1H), 2.00-1.35 (m, 9H), 1.1 (m, 2H).

2- [6- [2- (N-BOC-Piperidin-4-yl) ethyl} - (3,4,5,6-tetrahydro-1,1-dioxo-2H-1,2-thiazin-2-yl) acetic acid of the acid (91)

To a stirred solution of 89 (0.44 g, 1.13 mmol) in acetone (10 mL) at 0 ° C was added three times Jonah reagent (1 mL). After 5 minutes Excess ionic reagent is removed by addition of isopropanol and the reaction mixture is diluted with H ₂ O. The reaction mixture is extracted with ethyl acetate, the organic layer is washed with brine, dried over MgSO _4, and concentrated. Flash chromatography (silica, 20: 1: 1 CH ₂ Cl ₂ / CH ₃ OH / AcOH) gives 91 as a light yellow oil.

TLC R _f = 0.45 (20: 1: 1 CH ₂ Cl ₂ / CH ₃ OH / AcOH).

2- [6- [2- (N-BOC-Piperidin-4-yl) ethyl] - (3,4,5,6-tetrahydro-1,1-dioxo-2H-1,2-thiazin-2-yl) acetyl Preparation of -3- (R) methyl-p-alanine ethyl ester (92)

Using the same procedure as in 55 out of 50, 54 (120 mg, 0.71 mmol) was added to 91 (98 mg, 0.24 mmol), and then obtained by flash chromatography (silica, ethyl acetate) as a yellow oil.

TLC R _f = 0.28 (ethyl acetate);

¹ H NMR (300NHz, COCl ₃ ) δ 7.08 (m, 2H), 4.37 (m, 1H),

4.08 (m, 2H), 3.89 (d, J = 17 Hz, 1H), 3.77 (d, J = 17 Hz,

1H), 3.60 (m, 1H), 3.15 (m, 1H), 2.88 (m, 1H), 2.68 (m,

2H), 2.52 (m, 2H), 2.20-1.00 (m, 13H), 1.27 (t, J = 7Hz,

3H), 1.25 (t, J = 7Hz, 3H).

2- [6- [2- (N-BOC-Piperidin-4-yl) ethyl] - (3,4,5,6-tetrahydro-1,1-dioxo-2H-1,2-thiazin-2-yl) acetyl Preparation of -3- (R) methyl-β-alanine (93)

Using the same procedure as in 56 out of 55, 92 (76 mg, 0.15 mmol) were obtained in the form of 93 yellow oils.

TLC R _f = 0.42 (20: 1: 1 CH ₂ Cl ₂ / CH ₃ OH / AcOH.

1 H NMR (300 MHz, CDCl ₃ ) δ 7.20 (m, 1H), 4.379 (m, 1H), 4.10 (q, J = 7Hz, 2H), 4.08 (m, 2H), 3.92 (d, J = 17 Hz, 1H), 3.74 (m, 1H), 3.53 (m, 1H), 3.15 (m, 1H), 2.90 (m, 1H), 2.77 (m, 2H), 2.59 (m, 2H), 2.10 (m, 1H), 2.10 (m, 12H), 1.27 (t, J = 7Hz, 3H), 1.25 (t, J = 7Hz, 3H).

2- [6- [2- (Piperidin-4-yl) ethyl] - (3,4,5,6-tetrahydro-1,1-dioxo-2H-1,2-thiazin-2-yl) acetyl-3- ( Preparation of R) -methyl-β-alanine (94)

Using the same procedure as in 57 of 56, 93 (61 mg, 0.12 mmol) were obtained in 94 as a white solid after flash chromatography (silica, 10: 1: 1 ethanol / NH ₄ 0H / H ₂ O). .

TLC R _f = 0.26 (10: 1: 1 Ethanol / NH ₄ O / H ₂ O);

⁴ Η NMR (300 MHz, D ₂ O) δ 4.08 (m, 1H), 3.71 (m, 2H), 3.40 (m, 1H), 3.29 (m, 2H), 3.14 (m, 2H), 2.83 ( m, 2H),

2.25 (m, 2H), 2.12 (m, 1H), 1.90-1.20 (m, 12H), 1.05 (t, J = 7Hz, 3H).

Preparation of 1- [2- (N-BOC-piperidin-4-yl) ethyl] -3-propanol-2-pyrrolidinone (95) (1.20 g, 3.6 mmol) in THF (3 mL) was added to borane-dimethyl sulfide (3.6 mmol) ) in 3 mL of THF at 0 ° C and stirred for 2.5 h. at 0 ° C. H ₂ O (0.15 mL), 10N aqueous was then added at 0 ° C

A solution of NaOH (0.65 mL), THF (1 mL) and C ₂ H ₅ OH (0.36 mL) was stirred and the temperature was raised to 23 ° C and 30% H ₂ O ₂ (0.36 mL) was added. This solution is heated to 55 ° C and stirred for 2 hours. The reaction mixture is then cooled, diluted with K ₂ CO ₃ and the organic layer is separated. After drying over MgSO ₄ , the solvent is removed. The resulting residue is purified by flash chromatography on silica gel eluting with 10% MeOH / EtOAc to give 95. R _f 0.3 (silica gel, 10% MeOH / EtOAc).

A solution of 1- [2- (N-BOC-piperidin-4-yl) ethyl] -3-propanoic acid 2-pyrrolidinone (96) (1.14 mmol) in acetone (5 mL) was cooled to -15 ° C and treated with Jon's reagent, until the amber color changes to brown. The reaction mixture was diluted with saturated NaHCO ₃ solution to pH 9 and extracted with EtOAc. The aqueous phase is acidified to pH 3 with KHSO ₄ solution and extracted with several portions of EtOAc. The combined organic extracts were dried over MgSO ₄ and removed by solvent removal to afford 96. R _f 0.3 (silica gel, 9/1/1 CH ₂ Cl ₂ / MeOH / HOAc).

[1- [2- (N-BOC-piperidin-4-yl) ethyl] -2-pyrrolidinone-3] propanoyl-p-alanine ethyl ester (97)

A solution of 96 (0.55 mmol), β-alanine ethyl ester HCl (0.825 mmol), N-methylmorpholine (1.65 mmol), CH ₃ CH (5 mL) at 23 ° C was treated with BOP (0.825 mmol) and the resulting solution was stirred for 48 h. hours. The reaction mixture was diluted with EtOAC, washed with 10% KHSO ₄ , brine, H ₂ O, brine, dried over MgSO _4, and the solvent removed. The resulting residue is purified by silica gel flash chromatography eluting with 30% acetone / hexane to give 97 as a clear oil.

R _f 0.25 (silica gel, 30% acetone / hexane).

[Cl- [2- (N-BOC-piperidin-4-yl) ethyl] -2-pyrrolidinone-3] propanoyl-p-alanine (98)

A solution of 97 (0.29 mmol), LiOH.H ₂ O (1.45 mmol), THF / MeOH / H ₂ O (1: 1: 1, 3 mL) was stirred for 1 h, then diluted with EtOAc and washed with 10% KHSO _4. , H ₂ O, brine and dried over MgSO ₄ . Removal of solvent gives 98 as an oil. R _f 0.5 (silica gel, 9/1/1 CH ₂ Cl ₂ / MeOH / HOAc).

[1- [2- (Piperidin-4-yl) ethyl] -2-pyrrolidinone-3] propanoyl-p-alanine.HCl (99) (0.18 mmol) in EtOAc (2 mL) was cooled to -78 ° C and exposed to gaseous HCl. After 0.5 hour, the solvent removed in the form of a white solid is obtained 99.R 12:22 _f (silica gel, 10/1/1 EtOH, NH ₄ OH, H ₂ O).

Preparation of N-BOC-4-piperidine methyl iodide (101)

Using the same procedure as in 10 out of 9, out of 100 (12.3 g, 57 mmol) (Carr et al., EP 317997).

May 31, 1989) after flash chromatography (silica gel, 15% EtOAc / hexane) gave 101.

R _f = 0.38 (10% EtOAc / hexane).

¹ H NMR (300 MHz, CDCl 3) δ 1.15 (m, 2H), 1.48 (s, 9H),

1.62 (m, 1H), 1.84 (m, 2H), 2.70 (m, 2H), 3.11 (m, 2H), 4.12 (m, 2H).

Preparation of N-BOC-piperidinomethyltriphenylphosphonium iodide (102)

A solution of 101 (5.1 g, 15.7 mmol), CH ₃ CN (75 mL) and triphenylphosphine (4.5 g, 17.32 mmol) was heated at 80 ° C for hours. The cooled reaction mixture is concentrated, the remaining oil is mixed with ether and evaporated in vacuo to give 102 as yellow foam.

¹ H NMR (300 MHz, CDCl ₃ ) δ 7.31 (m, 15H), 4.10 (m, 2H),

3.97 (m, 2H), 2.69 (m, 2H), 2.00-110 (m, 5H), 1.48 (s,

9H).

Preparation of 3- [2- (N-BOC-piperidin-4-yl) ethylene] -2- (methoxy) pyridine (104) into a suspension of 102 (5.0 g, 8.5 mmol) in THF at 0 ° C was added dropwise to NaN (TMS) ₂ ( 9.4 mL, 9.4 mmol) solution. After 15 minutes 103 (0.97 g, 7.0 mmol) (Tetrahedron Letters (1988) 29,

773) dissolved in THF (3 mL) was added dropwise to the yellow / orange homogeneous reaction mixture. After 1 or. removable cooling bath and stir for 30 min. The reaction was quenched by the addition of H ₂ O and ether. The aqueous layer is re-extracted with ether. Combine the organic layers and wash with brine, dry over Na ₂ SO _4, and concentrate. Flash chromatography (silica, 15% ethyl acetate / hexane) gives 104 as a colorless oil.

TLC R _f = 0.15 (10% ethyl acetate / hexane)

100

NMR (300 MHz, CDCl ₃ ) δ 8.09 (dd, J = 5 and 2 Hz, 1H),

7.42 (dd, J = 7 and 2 Hz, 1H), 6.87 (dd, J = 7 and 5 Hz, 1H), 6.37 (d, J = 12 Hz, IH), 5.58 (dd, J = 12 and 10 Hz, 1H), 4.10 (m, 2H), 3.95 (s, 3H), 2.70 (m, 2H), 2.50 (m, 1H), 1.80- 1.20 (m, 4H), 1.47 (s, 9H).

Preparation of 3- [2- (N-BOC-piperidin-4-yl) ethyl] -2-methoxy-pyridine (105) -

A solution of 104 (1.4 g, 4.3 mmol), ethyl acetate (22 mL) and 10% Pd / C (0.27 g, 20% wt) in hydrogen (1 atm) was stirred at room temperature for 4 hours. The reaction mixture is filtered through a pad of celite, and the filtrate is concentrated to give 105 as a colorless, opaque oil.

TLC R _f = 0.17 (10% ethyl acetate / hexane)

¹ H NMR (300 MHz, CDC1 ₃ ) δ 8.02 (dd, J = 5 and 2 Hz, IH), 7.36 {dd, J = 7 and 2 Hz, 1H), 6.80 (dd, J = 7 and 5 Hz, IH), 4.10 (m, 2H), 3. 94 (s, 3H), 2.69 (s, 3H), 2.58 (m, 2H), 1.72 (m, 2H), 1. 55-1.35 (m, 3H), 1.48 (s, 9H), 1.13 (m,

2H).

Preparation of 3- [2- (N-BOC-piperidin-4-yl) ethyl] -2-pyridon-1-yl (106)

To a solution of 105 (0.81 g, 2.5 mmol) in dry CH ₃ CN (13 mL) was added NaI (0.94 g, 6.3 mmol), followed by chloromethylsilane (0.80 mL, 6.3 mmol), the resulting opaque yellow solution was heated at 60 ° C. hours. After cooling the reaction mixture quenched with methanol (50 mL), stirred 10 min. and concentrated. The residue was dissolved in dioxane / H ₂ 0 (6.7 ml / 3.3 ml) and treated with IN NaOH (5.0 ml, 5.0 mmol). The solution was cooled to 0 ° C, add BOC ₂ O (1.1 g, 5.0 mmol) and detach the cooling bath.

The contents of 101 are stirred overnight and the residue obtained after evaporation of dioxane is diluted with H ₂ O (10 mL) and ethyl acetate (50 mL) and acidified with 20% KHSO ₄ to pH ~ 1.0. The organic layer was separated, washed with H ₂ O and brine, dried (MgSO ₄ ), and concentrated. Flash chromatography (silica, 2: 1 CH ₂ Cl ₂ / acetone) gives 106 as a pale yellow solid.

PSC R _f = 0.19 (2: 1 CH ₂ Cl ₂ / acetone);

^X H NMR (400 MH z, CD Cl ₃ ) δ 7.30 (m, 3H), 6.23 (t, J = 7 Hz, 1H), 4.04 (m, 2H), 2.69 (m, 2H), 2.57 (m, 2H), 1.74 (m, 2H), 1.56 (m, 2H), 1.48 (m, 1H), 1.47 (s, 9H), 1.16 (m,

2H).

(Preparation of 3- [2- (N-BOC-piperidin-4-yl) ethyl] -2-pyridon-1-yl) acetate (107)

To a stirred solution of 106 (0.44 g, 1.4 mmol) in dry DMF (7.2 mL) at 0 ° C was added NaN (TMS) ₂ (2.1 mL, 2.1 mmol), ΙΜ / hexane). After 30 minutes, ethyl bromoacetate (0.79 mL, 7.2 mmol) was added dropwise to the reaction mixture. After an additional hour, the reaction mixture was diluted with ethyl acetate and washed with H ₂ O.5% KHSO ₄ , saturated NaHCO _3, and brine, dried over MgSO _4, and concentrated. Flash chromatography (silica, 45% ethyl acetate / hexane) gives 107 as a colorless oil.

PSC R _f = 0 .33 (60% ethyl acetate / hexane) ; δ 7.20 (dd, J = 6 and 1 Hz, 1H), 7.11 (dd, J = 7 and 2 Hz, 1H), 6.1! 5 (t, J = 7 Hz, 1H) , 4.62 (s, 2H), 4.24 (q, J = 7 Hz, 2H), 4.04 (m, 2H) , 2.69 (m, 2H), 2.55 (m, 2H), 1.72 (m, 2H), 1.52 (m, 2H) , 1.78 (m, 1H), 1.48 (s, 9H), 1 .13 (m, 2H).

Preparation of 102 (3- [2- (N-BOC-piperidin-4-yl) ethyl] -2-pyridon-1-yl} acetic acid (108)

Using the same procedure as in 56 out of 55, 108 out of 107 (0.51 g, 1.3 mmol) are obtained in the form of colorless foam.

TLC R _f = 0.58 (9: 1: 1 CH ₂ Cl ₂ / CH ₃ OH / HQAc).

Preparation of {3- [2- (N-BOC-piperidin-4-yl) ethyl] -2-pyridon-1-yl] acetyl-3 (R) -methyl-p-alanine ethyl ester (109)

Using the same procedure as in 16 out of 15, 109 (0.15 g, 0.41 mmol) after flash chromatography (silica, ethyl acetate) are obtained in the form of 109 waxes.

TLC R _f = 00:38 (ethyl acetate)

¹ H NMR (400 MHz, CDC1 ₃ ) δ 7.24 (dd, J = 7 and 2 Hz, 1H), 7.21 {dd, J = 7 and 2 Hz, 1H), 7.07 (pld, J = 8 Hz, 1H), 6.19 (t, J = 7Hz, 1H), 4.51 (s, 2H), 4.30 (m, 1H), 4.08 (q, J = 7 Hz, 2H), 4.08 (m, 2H), 2.69 (m, 2H), 2.55 (m, 2H), 1.72 (m, 2H), 1.51 (m, 2H), 1.45 (s, 9H), 1.43 (m, 1H), 1. 23 (t, J = 7Hz, 3H), 1.22 (d, J = Ί Hz, 3H), 1.13 (m, 2H)

Preparation of {3- [2- (N-BOC-piperidin-4-yl) ethyl] -2-pyridon-1-yl} acetyl-3 (R) -methyl-p-alanine (110)

Using the same procedure as in 56 out of 55, 110 (0.19 g, 0.41 mmol) are obtained in the form of 110 as a colorless oil. PSCR _f = 0.59 (9.5: 0.5: 0.5) CH ₂ Cl ₂ / CH ₃ OH / HOAc).

(Preparation of 3- [2- (Piperidin-4-yl) ethyl] -2-pyridon-1-yl] acetyl-3 (R) methyl-β-alanine, (III)

103 Solution, consisting of (0.18 g, .0. , 4 0 mmo1), CH ₂ Cl ₂ (2.0 ml), anisole (86 μΐ, 0.80 mmol) and trifluoro acetic acid (2.0 mL) at -15 ° C at temperature stirring 15 mins Yellow reactions blend concentrated and the rest trifluoroacetic acid acid is removed aceotropic distillation with toluene help. After high-speed chromatography (silicon oxide, 10 / 1.3 / 1.3 Ethanol NH ₄ OH / H ₂ O) is obtained 111 white amorphous materials in the form of. TLC R _f = 00:17 (10: 1: 1 ethanol NH ₄ OH / H ₂ O); ^X H NMR (400 MHz, D ₂ O) δ 7.57 (dd, J = 7 and 1 Hz, 1H), 7.4 9 (dd, J = 7 and 1 Hz, 1H), 6.51 (t, J = 7 Hz, 1H),

4.66 (m, 2H), 4.20 (m, 1H), 3.42 (m, 2H), 2.97 (m, 2H), 2.59 (m, 2H), 2.46 (d, J = 14 and 6 Hz, 1H), 2.32 (dd,

J = 14 and 7 Hz, 1H), 2.00 (m, 3H), 1.42 (m, 2H).

Preparation of N-Cbz-4-piperidinethanol (112)

To a stirred solution of 4-piperidinethanol 8 (15 g, 0.12 mmol), THF (500 mL) and diisopropylethyleneaminate at 0 ° C was added benzyl chloroformate (16.5 mL, 0.12 mmol) and incubated for 1 h. At 0 ° C. The reaction mixture was then diluted with ethyl acetate and washed with H ₂ O (2 times, 10% KHSO ₄ ) and brine, dried over MgSO ₄ and concentrated to give 112 as a colorless oil.

TLC R _f = 0.60 (ethyl acetate);

¹ H NMR (300 MHz, CDCl ₃ ) δ 7.35 (m, 5H), 5.13 (s, 2H),

4.18 (m, 2H), 3.70 (sq, J = 7 Hz, 2H), 2.80 (m, 2H),

1.80-1.50 (m, 5H), 1.19 (m, 2H).

Preparation of N-Cbz-4-piperidine ethyl iodide (113)

104

Using the same procedure as in 10 out of 9, 113 (30.6 g, 0.12 mol) are obtained in the form of a colorless oil after flash chromatography (silica, 10% ethyl acetate / hexane).

1- [2- (N-Cbz-piperidin-4-yl) ethyl] - (2-pyrrolidinone) (114)

To a stirred solution of 2-pyrrolidinone (11.0 g, 0.13 mmol) in dry DMF (500 mL) was added dropwise NaN (TMS) ₂ (129 mL, 129 mmol, 1M / THF). After 10 minutes. of iodide 113 (24.1 g,

64.6 mmol) solution in DMF (50 mL). After 1 hour, the reaction mixture was diluted with ethyl acetate and washed with H ₂ O, saturated NaHCO ₃ , brine, dried (MgSO ₄ ), and concentrated. Flash chromatography (silica, ethyl acetate) gives 113 as a colorless oil.

TLC R _f = 00:16 (ethyl acetate);

^Τ Η NMR (300 MHz, CDC1 ₃ ) δ 7.40-7.20 (m, 5H), 5.12 (s, 2H) , 4.19 (m, 2H) , 3.34 (m, 2H), 2.77 (m, 2H), 2.40 (m, 2H) , 2.04 (m, 2H) , 1.74 (m, 2H), 1.47 (m, 3H), 1.15 (m,

2H).

Preparation of 1- [2- (N-Cbz-piperidin-4-yl) ethyl] -3-propen-2-yl- (2-pyrrolidinone) (115)

To a stirred solution of 114 (8.0 g,

24.2 mmol) and THF (180 mL) were added dropwise to LDA (50 mL, 25 mmol, 0.5M / THF) at -78 ° C. After 15 minutes Allyl bromide (2.3 mL, 26.6 mmol) was added. After 1 hour, the reaction mixture was stirred for 10 min. is heated to 0 ° C. The reaction mixture was then diluted with ethyl acetate and washed with 5% KHSO ₄ and brine. dried over MgSO ₄ and concentrated. After a quickLT 3284 B

105% chromatography (silica, 50% ethyl acetate / hexane) gives 115.

TLC R _f = 0.65 (ethyl acetate);

⁷ Η BMR (300 MHz CDC1 _3; > δ 7.35 (m, 5H), 5.80 (m, 1H), 5.12 (m, 2H) , 4.18 (m, 2H) , 3.33 (m, 4H), 2.80 (m, 2H), 2.56 (m, 2H) , 2.20 (m, 2H) , 1.78 (m, 2H), 1.48 (m, 3H), 1.20 (m, 2H) •

Preparation of 1- [2- (N-Cbz-piperidin-4-yl) ethyl] -3-acetic acid-2-pyrrolidinone) (116)

Using the same procedure as for 15 out of 14, from 115 (4.7 g, 12.7 mol) after flash chromatography (silica, 9.5: 0.5: 0.5 CH ₂ Cl ₂ /

CH ₃ OH / HOAc) is obtained in the form of 116 light brown oil.

PSC R _f = 0.63 (9.5: 0.5: 0.5 CH ₂ Cl ₂ / CH ₃ OH / HOAc);

¹ H NMR (300 MHz, CDCl ₃ ) δ 7.37 (m, 5H), 5.15 (s, 2H),

4.18 (m, 2H), 3.37 (m, 4H), 3.00-2.30 (m, 5H), 1.77 (m, 2H), 1.46 (m, 3H), 1.18 (m, 2H).

Preparation of 1- [2- (N-Cbz-piperidin-4-yl) ethyl] -2-pyrrolidinone 3] acetyl-2-aminoethylphosphonic acid (117)

A solution of 116 (388 mg, 1.0 mmol), Et ₃ N (0.14 mL, 1.0 mmol) and dry dioxane (5 mL) was treated with isobutyl chloroformate (0.13 mL, 1.0 mmol) at 0 ° C. The reaction mixture is heated until the dioxane melts. A solution of 2 aminoethylphosphonic acid (125 mg, 1.0 mmol), Na ₂ CO ₃ (106 mg) and H ₂ O (2 mL) was added to the reaction vessel at room temperature. After 4 hours, dioxane

106 was evaporated and the residue acidified with IN HCl. The mixture was washed with ethyl acetate (3x).

The aqueous layer is concentrated to dryness to give crude 117.

TLC R _f = 0.15 (9: 1: 1 CH ₂ Cl ₂ / CH ₃ OH / HOAc).

Preparation of 1- [2- (piperidin-4-yl) ethyl] -2-pyrrolidinone-3] acetyl-2-aminoethylphosphonic acid (117)

The crude phosphonic acid 117 (464 mg) was dissolved in 10% HOAc / CH ₃ OH (20 mL), then Pd (OH) ₂ (50 mg) was added and the mixture was hydrogenated overnight in a Parr apparatus (50PSI). After 20 hours, the reaction mixture is filtered through a pad of celite and the filtrate is concentrated. Flash chromatography (silica, 8: 1: 1 CH ₃ OH / H ₂ O / NH ₄ OH) affords 117 amorphous material.

TLC R _f = 0.14 (8: 1: 1 CH ₃ OH / H ₂ O / NH ₄ OH);

³ H NMR (300 MHz, D ₂ O δ 3. 40 (m, 8H), 3.18 (m, 1H), 2.92 (m, 3H), 2.60 (m, 1H), 2.38 (m, 1H), 2.26 (m, 1H), 1.97 (m, 2H), 1.79 (m, 3H), 1.56 (m, 2H), 1.40 (m, 2H). (4-N-Cbz -amino) cyclohexane carbocyclic of acid

obtaining the methyl ester (121)

To a suspension of 120 (17.5 g, 0.12 mol) (150 mL) in CH ₃ OH at -10 ° C for 5 min. Add thionyl chloride (13.4 mL, 0.18 mol) in portions and remove the cooling bath. After 16 hours, the resulting solution was concentrated, the residue was dissolved in DMF (150 mL), cooled to 0 ° C and treated first with N (i-Pr) ₂ Et (52 mL, 0.3 mol) followed by benzyl chloroformate (18.6 mL, 0.13). mol). Remove the cooling bath, incubate the reaction mixture for 24 hours, and concentrate. Remain

107 was diluted with ethyl acetate and washed with dilute NaHCO _3, 5% KHSO ₄ and brine, dried over MgSO ₄ and concentrated. Flash chromatography (silica, 20% ethyl acetate / hexane) gave a crystalline wax-like material 121.

TLC R _f = 0.34 (30% ethyl acetate / hexane);

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.38 (m, 5H), 5.09 (pis, 2H),

4.68 (m, 0.5H), 4.59 (m, 0.5H), 3.72 (m, 0.5H), 3.68 1.50 (s, (m, 1.5H), 8H). 3.50 (m, 0.5H), 2.45 (m, IH), 2.13-

Preparation of (4-N-Cbz-amino) [1- (propen-3-yl)] -cyclohexane carboxylic acid methyl ester (122)

To a stirred solution of 121 (1.0 g, 3.4 mmol) in dry THF (17.2 mL) was added dropwise LDA (4.3 mL, 8.6 mmol, 2.OM) at -78 ° C. The temperature of the cooling bath is raised to -40 ° C. After stirring for 15 min. At -40 ° C, the reaction mixture was again cooled to -78 ° C and treated sequentially with 1,3-dimethyl-3,4,5,6-tetrahydro-2- (1H-pyrimidinone) (2.0 mL, 17 mmol ) and allyl bromide (0.44 mL, 5.1 mmol). After 2 hours, the reaction mixture was diluted with ethyl acetate, washed with H ₂ O), saturated NaHCO _3, and brine. Dry over MgSO ₄ and concentrate to give a yellow oil which is purified by flash chromatography (silica, 20% ethyl acetate / hexane) to give a crystalline waxy material 122.

TLC R _f = 0.31 (20% ethyl acetate / hexane);

1 H NMR (400 MHz, CDCl ₃ ) δ 7.34 (m, 5H), 5.67 (m, 1H),

5.10 (s, 2H), 5.05 (m, 2H), 4.54 (m, 1H), 3.67 (s, 3H),

3.48 (m, 1H), 2.21 (m, 2H), 2.00-1.00 (m, 8H).

108

Preparation of 4-Amino, 1- (propen-3-yl)] cyclohexane carboxylic acid methyl ester (123)

Trifluoroacetic acid (19 mL) at 0 ° C to a mixture of 122 anisole (0.41 mL, 3.8 mmol). the mixture becomes homogeneous and left to stir overnight.

(0.63 g, 1.9 mmol) and after 5 min. reactions after removing the cooling bath,

Concentration affords 123 as a yellow oil after flash chromatography (silica, 17: 1: 1 ethanol / NH ₄ 0H / H ₂ 0).

TLC R _f = 0.53 (30% ethyl acetate / hexane).

Preparation of [2-Aza-3-oxo- [2.2.2] bicyclooct-4-yl] prop-3-ene (124)

A solution of 123 (0.39g, 1.97 mmol) in toluene (10 mL) in a sealed ampoule was heated at 170 ° C overnight. The cooled dark reaction mixture was purified by flash chromatography (silica, 85% ethyl acetate / hexane) to give 124 as a brown oil.

TLC R _f = 0.30 (85% ethyl acetate / hexane);

¹ H NMR (400 MHz, CDCl ₃ ) δ 6.08 (m, 1H), 5.88 (m, 1H),

5.04 (m, 2H), 3.61 (m, 1H), 2.35 (d, J = 7Hz, 2H), 1.801.50 (m, 8H).

(Preparation of [2- (N-BOC-piperidin-4-yl) ethyl] -2-aza-3-oxo [2.2.2] bicyclooct-4-yl] prop-3-ene (125)

Using the same procedure as for 37a from

36a, of 124 (0.18 g, 1.1 mmol) after high-speed chromatoLT 3284 B

109 (silica, 40% ethyl acetate / hexane) yields 125 (0.45 g, 100%).

TLC R _f = 0.36 (40% ethyl acetate / hexane);

^X H NMR (400 MHz, CDCl ₃ ) δ 5.87 (m, 1H), 5.05 (m, 2H), 4.08 (m, 2H), 3561 (m, 1H), 3.40 (t, J = 8 Hz, 2H), 2.67 (m, 1.47 (s, 2H), 2.34 (d, J = 7Hz, 2H), 1.75 9H), 1.12 (m, 2H). -1.35 (m, 13H),

Preparation of 3- (1- [2- (N-BOC-piperidin-4-yl) ethyl] -2-aza-3-oxo [2.2.2] bicyclooct-4-yl] acetic acid (126)

Using the same procedure as 15 of 14, 125 (0.42 g, 1.1 mmol) of 126 (0.34 g, 76%) are obtained in the form of white sticky foam.

(Preparation of [2- (N-BOC-piperidin-4-yl) ethyl] -2-aza-3-oxo [2,2,2] bicyclooct-4-yl] acetyl] -β-alanine (127)

Using the same procedure as in 16 out of 15, 127 (0.10 g, 55%) are obtained from 126 (0.14 g, 0.35 mmol) after flash chromatography (silica, 6: 1 CH ₂ Cl ₂ / acetone).

PSC R _f = 0.52 (CH ₂ Cl ₂ / acetone);

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.90 (m, 1H), 4.04 (m, 2H),

3.56 (m, 1H), 3.42 (m, 4H) 2.67 (m, 2H), 2.42 (d, J = 7Hz, 2H), 1.76-1.34 (m, 13H), 1.46 (s, 18H), 1.12 ( m, 2H).

([2-Piperidin-4-yl) ethyl] -2-aza-3-oxo [2.2.2] bicyclooct-4-yl] acetyl] -alanine (128)

Using the same procedure as for 41a from

40a, of 127 (0.10 g, 0.19 mmol) after high-speed chromiumLT 3284 B

110 tography (silica, 10: 1.2: 1.2 ethanol /

NH ₄ OH / H ₂ O) is obtained in 128 (29 mg, 39%) as a friable solid.

TLC R _f = 0.17 (10: 1: 1 ethanol / NH ₄ O / H ₂ O;

¹ H NMR (400 MHz, D ₂ O) δ 3.78 (m, 1H), 3.50-3.35 (m, 6H), 2.97 (m, 2H) 2.48 (s, 2H), -2.38 (t, J = 7 Hz) , 2H), 2.02 (m, 2H), 1.78 (m, 6H), 1.58 (m, 5H), 1.42 (m, 2H).

The methods of the present invention include those known in the literature for use in the preparation of the compounds of the invention in the synthesis of amidated guanidine derivatives and amidines of the corresponding nitriles. Guanidine compounds may be prepared from amines by reaction with 3,5-dimethylpyrazole-1-carboxamidine nitrate (Meth. Enzymol. 25b, 558, 1972). Amidines may be prepared from the corresponding nitriles by methods known to those skilled in the art using the procedures provided by Boere, R. T, et al. J.Organoment.Chem., 331 (2), 161-7, 1987; and Fuchs, R., Tetrahedron, 29 (14), 2147-51, 1973.

The materials listed in Table 1 are examples of compounds that may be obtained using the methodology of the present invention.

Table 111

O

R ¹ -CH ₂ -C-N 11 -R ²

f f .; O

Tables 112 (continued).

r 'o

sOj-ch, * y-j

R ²

CH ₂ Ph ^{o <} c-ch-co, h

I

-CH-CH ₂ CO ₂ H f ai

CH, o

r 7 — CK ₂ - «r · '-

CH-CH, -CC, H oV,

CIIj

-CH ₂ -C! L-COJi

O

II

C-Pho I

-CH-CH ₂ -CO ₂ K

KH

II

H ₂ II -C- (CH ₂ ) ₃

I · 'o

II

C-Pho-Luu I

-C H-C H,

O

CO ₂ ii

113

In vivo activity

The following experimental protocols were used in the present invention to determine the activity of fibrinogen receptor antagonists in vivo in dogs.

The non-purebred dog of any breed (weighing 7.5 to 11.5 kg) is comfortably anchored in nylon loops that uncover the limbs and fix the dog permanently. Using a plastic syringe containing 0.5 ml of 3.8 citrate, 5 ml of blood is drawn from the subcutaneous or maxillary vein through a 19-gauge inlet flap. Take an additional 1 ml of citrate to determine the total number. Ex vivo platelet aggregation is performed using ADP (10 μΜ) and collagen (10 μg / ml) containing 1 μΜ epinephrine as agonists. Platelet-rich plasma (TPP) is obtained by centrifugation of the blood at 150 x g for 5 min. Platelet counts are standardized to 200,000 / mm ³ by diluting plasma with low platelet counts.

blood with platelets

Oral administration of the drug is carried out either in the form of a gelatin capsule or by gastric lavage. Using the gastric lavage method, the dog is administered 5 ml of the solution via the feeding tube. Blood samples for ex vivo platelet aggregation are taken at the following time intervals: 0, 20, 40, 70, 90, 150, 200, 250, 300, 350, 480 min, 24 h, 30 h, 48 h. Platelet-poor plasma is left (at each time point) and frozen for drug determination. Three additional blood samples are taken at 30, 55 and 110 min. after administration to determine plasma levels of drugs.

For intravenous infusion, blood samples for ex vivo platelet aggregation are taken at the following time intervals: 0, 30, 45, 60, 90 and 120 min. infusions

At 114 and 2.5, 15, 30, 60, 90, 120, 180, 240, 300, 360 and

420 min. after the infusion. Platelet-poor plasma is left (at each time point) and frozen for drug determination. Drugs are infused at a constant rate of 0.1 ml / min. speed.

In the case of a rapid intravenous infusion, 5 ml of the solution is administered rapidly. Blood samples for ex vivo platelet aggregation are taken at the following time intervals:

0, 1, 5, 10, 15, 30, 45, 60, 90, 120, 180 and 240 min.

after the bolus. Platelet-poor plasma is left (at each time point) and frozen for drug determination.

In both the oral and intravenous groups, blood sampling times are recorded at each time point to determine plasma levels of drugs.

A compound of formula

HN

was determined according to the methodology above.

Platelet aggregation inhibition (% of control) induced by intravenous infusion at 10 µg / kg / min and 25 µg / kg / min was detected. Inhibition was practically complete during the infusion and partial inhibition was observed several hours after completion of the infusion.

115

Platelet aggregation inhibition (% of control) induced by a single (2 mg / kg) oral administration was found. The% inhibition of platelet aggregation was greater than 50% at 8 hours. over time.

Claims (11)

1. A fibrinogen receptor antagonist represented by Formula I: ^0R X- (CH ₂ ) _m -Y- <CH ₂ ) _k -C-NII-CH-CH-Z wherein X is NH NH NH -NR ⁷ R ³ , -nh-c-nr ⁷ r ³ , -c-nr ⁷ r ³ , r ⁷ -c-nh-, H, N —— <S or Oh ₂ n-ch ₂ wherein D wherein D = -C-, -S (O) g, or -O-; R * NH '-A «Ί, / Χ ₄ <A /' \ / R ⁴ . Cch ₂ ) _. ^B ~ \ '^ R < or X wherein A = N and B = -CH ₂ , or -CH- and B = NR ⁷ ; 117 where n = 0-5; O ₂ z is -CO ₂ r ₂ ; _J_ _oe _ £ _ _oh • OR ² ; R ¹ ®; wherein R ¹⁰ is C 1-6 alkyl, aryl, arylC 1-6 alkyl; U is -CH-, -C-, or -N-; V is -CH-, -C-, or -N-; modified or selected from iodine, cyano, R and R are independently hydrogen, aryl, wherein aryl is defined as a mono- or polycyclic aromatic system consisting of 5 or 6 membered rings having 0, 1, 2, 3 or 4 heteroatoms selected from nitrogen, oxygen and sulfur; and phenyl, unsubstituted by one or more groups, hydroxyl, fluorine, chlorine, trifluoromethyl, C 1-4 alkoxyl, C ₁ _ ₅ alkoxycarbonyl, C ₁ _ ₅ alkyl, aminoC ^ ₅ alkyl, hidroksikarbonilC ₀ _ ₅ alkyl or hidroksikarbonilC _{first 5} alkoxyl. bromine, C _x . ₅ alpha- ₁ carbonyl-oxy, ₀ _ C ₅ alkyl, substituted or unsubstituted by one or more groups selected from fluoro, chloro, bromo, iodo, hydroxy, C ^ alkylcarbonyl (C ₀ _ ₈₎ amino, aryl Ci-salkilkarbonil (c _{0th 8} alkyl) amino, aryloxy, C _{first 10} alkoxy, 118 C ₁ _ ₅ alkoxycarbonyl, C ₀ _ ₅ alkylaminocarbonyl, C _{x. 5} alkylcarbonyloxy, C ₃ _ ₈ cycloalkyl, aryl, oxo, amino, Cy.galkilo C ₁ _ ₃ alkylamino, aminoC _{first 3} alkyl, arylC ₀ _ ₅ alkylaminocarbonyl fenilC ₁ _ ₃ alkylamino, aminocarbonyl C ₀ . ₄ alkyl, C ^ alkylsulfonyl (C ₀ _ ₈ alkyl) amino, arylC ₀ s ₀ alkylsulfonyl (C _{0th 8)} amino or ILC _{0th 8} alkyl sulfonyl C ₀ . ₈ alkylsulphonylamino onyl, hidroksikarbonilC -5alkilo ₀ C _{first 8} alkoxycarbonyl (C _{0th 8} alkyl) amino, arylC _{0th 10} alkoxycarbonyl (C _{0th 8} alkyl) amino, C ₀ _ ₈ alkylaminocarbonyl (C _{0th 8} alkyl) amino, arylC ₀ _ ₈ alkylaminocarbonyl (C _{0th 8} alkyl) amino, C _{0th 8} alkylaminocarbonyloxy, or iC ₀ . ₁₀ a To laminate carbonyl oxy, C ₀ . ₈ alkylsulfonylamino (C _{0th 8} alkyl) amino, arylC ₀ _ ₈ alkylsulfonylamino (C _{0th 8} alkyl) amino, C _{0th 8} alkylaminosulfonyl, or arylC ₀ . ₈ alkylaminosulfonyl; with the proviso that the carbon atom to which R or R ¹ is attached has only one heteroatom; R is hydrogen, C ₁ . ₁₂ alkyl substituted or unsubstituted with one or more 0 _χ . ₆ alkyl groups, o -CH ₂ 0CR ⁹ , R ⁹ 0 ⁹ , -CHOCR ⁹ wherein R ⁹ = branched or unbranched C _{x 6} alkyl or phenyl and wherein R ⁹ when more than one may be the same or different; R ⁷ , R ³ and R ⁴ are independently hydrogen 119 C ₁ . ₁₂ alkyl substituted or unsubstituted with one or more C 1-4 alkyl groups or arylC ₀ . ₄ alkyl, or cyano, provided that when R ⁷ and R ³ are independently cyano, X is k is 1-4; m is 1-4; p is 1-6; q is 0-2; or pharmaceutically acceptable salts thereof, or optical isomers thereof. 2. The fibrinogen receptor antagonist of claim 1, wherein the formula I wherein: X is CCH ₂ ) p I 120 or H _{2 is} N-CH, or wherein n is 1, 2, or 3; Z is CO ₂ R ² ; R and R ¹ are, independently selected from phenyl, thiophene, imidazole, naphthyl, indole, indazole, tionafteno unsubstituted or substituted by hydroxyl, halogen, hidroksikarbonilC ₀ _ ₅ alkyl, C _{first 3} alkyl, substituted or unsubstituted by one or more groups selected from aryl, aryloxy, (©. ^ Alkoxy, C ₀ _ ₅ alkylaminocarbonyl, arylC ₀ _ ₅ alkylaminocarbonyl, hydrogen, C ₀ _ ₆ alkyl, substituted or unsubstituted by one or more groups , 121 selected from halogen, hydroxyl, C ₁ . ₆ alkylsulfonylamino, arylC ₀ . ₆ alkylsulfonylamino, C ^ galkilsulfonilo, arylC ₀ _ ₆ alkylsulphonyl, C _{first 5} alkylcarbonylamino, arylC ₁ _ ₅ alkylcarbonylamino, aryloxy, C ^ ujalkoksi C _{first 5} alkoxycarbonyl, C ₀ _ ₅ alkylaminocarbonyl, C _{first 5} alkylcarbonyloxy, C ₃ _ ₈ cycloalkyl, aryl, oxo, amino, _{first 6} alkyl, C ₁ _ ₃ alkylamino, aminoC _{first 3} alkyl, arylC ₀ _ ₅ alkylaminocarbonyl fenilC _{first 3} alkylamino, aminocarbonylC ₀ . ₄ alkyl, or hydroxycarbonylC _{Q. 5} alkyl, provided that the carbon atom to which R or R ¹ is attached has only one heteroatom; R ² is hydrogen, C ₁ . ₁₂ alkyl, substituted or unsubstituted by one or more C ₁ _ ₆ alkyl groups, 0 R ⁹ 0 CHo CHo 0 ·> _Q I II d -CH ₂ 0CR ⁹ , -CB 0CR ^9, or -C-0 _: -CR ^y where R = branched or unbranched C ₁ . ₆ alkyl, or phenyl; R, R and R are independently hydrogen or C 1-4 alkyl, substituted or unsubstituted with one or more C _{1 's} . ₆ alkyl groups; k is 1-4; m is 1-4; p is 1-3; q is 0 or 2; Or pharmaceutically acceptable salts thereof or optical isomers thereof. 3. The fibrinogen receptor antagonist of claim 1, wherein the fibrinogen receptor antagonist is as defined in claim 1 X is (CH ^ p ' R -NN k ₃ n-ch ₂ o K where D = -O-, -S-, or -CZ is CO ₂ R ' Y is R O R or R wherein n is 1, 2 or 3; 123 R and R ^{1 are} independently selected from phenyl, thiophene, imidazole, naphthyl, indole, indazole, thionaphthene, substituted or unsubstituted hydroxyl, halogen, hydroxycarbonylC ₀ . ₅ alkyl, C ₁ . ₃ alkyl substituted or unsubstituted with one or more groups selected from aryl, aryloxy, C _x . ₁₀ alkoxy, C ₀ _ ₅ alkylaminocarbonyl, arylC _{0th 5} alkylaminocarbonyl, hydrogen, C ₀ _ ₆ alkyl substituted by one or more groups selected from halogen, hydroxy, C ₁ _ ₅ alkylcarbonylamino, arylC _{first 5} alkylcarbonylamino, aryloxy, C ₁ _ ₅ alkoxycarbonyl, C ₀ _ ₅ alkylaminocarbonyl, C ^ alkylcarbonyloxy, C _{3rd 8} cycloalkyl, aryl, oxo, amino, C _x . ₆ alkyl, C ₁ . ₃ alkylamino, aminoC ₁ . ₃ alkyl, arylC _0-5 alkylaminocarbonyl, fenilC _{first 3} alkylamino, aminokarbonilC ₀ _ ₄ alkyl or hidroksikarbonilC _{0 5} alkyl, with the proviso that the carbon atom is connected to R or R ¹ having only one hetero atom; R ² is hydrogen; R ⁷ , R ³ and R ^{4 * *} are hydrogen; k is 1-2; m is 1-4; p is 2-4; or pharmaceutically acceptable salts thereof, or optical isomers thereof. A compound according to claim 1 for use in inhibiting fibrinogen binding to platelets, inhibiting platelet aggregation, treating thrombus or embolus formation, or preventing thrombus or embolism formation in a mammal. 124 5. The compound of claim 1, wherein the compound is selected from the group consisting of: OH, HN 2 « H. SCH, ⁿ ^ \ h'V ^C00h > KN HO _NH - ^ COOH 125 126 O H J H O Or a pharmaceutically acceptable salt thereof. 6. A compound according to claim 5, which is sch ₃ on or a pharmaceutically acceptable salt thereof. 7. The compound of claim 5, wherein it is 128 or a pharmaceutically acceptable salt thereof. 8. A compound according to claim 5 which is or a pharmaceutically acceptable salt thereof. 9. A compound according to claim 5, which is or a pharmaceutically acceptable salt thereof. 10. A compound according to claim 5 which is Or pharmaceutically acceptable salts thereof. A compound according to claim 1, wherein the compound is different or pharmaceutically acceptable salts thereof.