ES2600320B1 - HETEROCYCLIC COMPOUNDS AS ANTAGONISTS OF TRPM8 CHANNELS AND THEIR USES - Google Patents

Abstract

Heterocyclic compounds as antagonists of TRPM8 channels and their uses. # The invention relates to derivatives {be} -lactamics, capable of blocking the activation of thermoreceptors, in particular TRPM8 channels and their uses as therapeutic agents for the treatment of diseases of the system respiratory (cough, asthma, etc.), of the skin (pruritus, atopic dermatitis, allergen, psoriasis, etc.), of the eye (dry eye syndrome and excessive tearing), derived from cancer chemotherapy (cold allodynia) , as well as cancer (melanoma, prostate, kidney, breast, pancreas, osteosarcoma, etc.), among others.

Description

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HETEROCICLICAL COMPOUNDS AS TRPM8 CHANNEL ANTAGONISTS

AND ITS USES

DESCRIPTION

The present invention relates to heterodclic compounds, which are p-lactam derivatives, capable of blocking the activation of thermoreceptors, in particular TRPM8 channels.

STATE OF THE TECHNIQUE

Thermosensory channels, a subfamily within transient potential receptors (TRP), are activated by changes in temperature, voltage, pressure, acids, endogenous ligands and different products of natural origin. These channels, known as thermoTRPs, cover the entire temperature spectrum, from harmful cold (<15 ° C) to harmful heat (> 42 ° C). In addition, the alterations in these channels contribute to the thermal hypersensitivity associated with certain painful episodes, in addition to being involved in the pathophysiology of various diseases, such as inflammation, lung disorders, gastrointestinal hypersensitivity, skin sensitivity, neurodegeneration and cancer.

TRPM8 receptors are channels permeable to Ca2 + that were initially identified in prostate tumor cells, but which are also present along the male urogenital tract, arteries, and pulmonary epithelial cells. These channels are expressed in primary sensory neurons of the skin and mucous membranes, acting as low temperature sensors (10-33 ° C) and chemicals such as menthol and icilin. Under pathological conditions, there is an increasing number of experimental evidence confirming the anomalous overexpression of TRPM8 channels in sensory neurons after injury or inflammation, as well as their participation in allodynia and cold hyperalgesia. TRPM8 is also expressed in corneal afferent neurons involved in the regulation of ocular surface moisture.

Among TRPM8 antagonists, different families have been reported to have benzothiophene, benzimidazole and arylglycine residues as the central skeleton (Calvo et al 2012 Bioorganic & Medicinal Chemistry Letters 22: 1903-1907; Matthews et al 2012

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Bioorganic & Medicinal Chemistry Letters 22: 2922-2926; Parks et al 2011 Journal of Medicinal Chemistry 54: 233-247 ;. Zhu et al 2013 Bioorganic & Medicinal Chemistry Letters 23: 2234-2237.). Some of these compounds have shown excellent "in vitro" and "in vivo" activities in animal models of inflammatory and neuropathic pain. The compound JNJ41876666, derived from benzothiophene, has been used, together with another antagonist, AMTB, and RNAi to determine that inhibition of both the expression and function of the TRPM8 channels reduces the rate of prostate tumor cell proliferation (Valero et al., 2012 PLoS ONE 7 (12): e51825). The commercial antagonist N- (3-aminopropyl) -2 - [(3-methylphenyl) methoxy] -N- (2-

thienylmethyl) benzamide AMTB has also served to demonstrate that TRPM8 channel blockage is a new therapeutic opportunity for the treatment of hyperactive and painful bladder smdromes (Lashinger et al. 2008 American Journal of Physiology-Renal Physiology 295: F803-F810) .

DESCRIPTION OF THE INVENTION

The present invention provides heterocyclic compounds with very significant activity as blockers of thermoreceptors, in particular TRPM8. The blocking activity of the aforementioned channels, which these compounds have, allows them to be used as therapeutic agents for the treatment of diseases of the respiratory system (cough, asthma, etc.), of the skin (pruritus, atopic dermatitis, allergen, psoriasis, etc.) , of the eye (dry eye smdrome and excessive tearing) of cancer chemotherapy (cold allodynia), as well as cancer (melanoma, prostate, kidney, breast, pancreas, osteosarcoma, etc.).

Therefore, a first aspect of the present invention relates to a compound of formula (I) or any of its pharmaceutically acceptable salts or stereoisomer thereof (hereinafter compound of the invention):

image 1

) —Ra

R5

(I)

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where:

R1 is selected from (C1-C4) alkyl, substituted or unsubstituted, aralkyl (C1-C4), substituted or unsubstituted, heteroaralkyl (C1-C4), substituted or unsubstituted, aryl, substituted or unsubstituted, and heteroaryl , substituted or unsubstituted;

R2 is selected from a group -OR6 and -NR7R8;

R3 is selected from hydrogen and (C1-C4) alkyl, substituted or unsubstituted;

R4 is selected from hydrogen, (C1-C4) alkyl, substituted or unsubstituted, aralkyl (C1-C4), substituted or unsubstituted, heteroaralkyl (C1-C4), substituted or unsubstituted, aryl, substituted or unsubstituted, and -COR9;

R5 is selected from -NR11R10;

R8 is selected from hydrogen, (C1-C6) alkyl and aralkyl (C1-C4), heteroaryl, heteroarylalkyl (C1-C4) and -CH (R13) COOR12;

R6, R11, R13 and R14 are independently selected from hydrogen, (C1-C6) alkyl, substituted or unsubstituted, aralkyl (C1-C4), substituted or unsubstituted, heteroaralkyl (C1-C4), substituted or unsubstituted, and aryl, substituted or unsubstituted;

R7 and R12 are selected from hydrogen and (C1-C6) alkyl, substituted or unsubstituted; R9 is selected from -OR6 and -NR7R8;

R10 is selected from hydrogen, (C1-C6) alkyl, substituted or unsubstituted, and (C1-C4) aralkyl, substituted or unsubstituted, COOR14; and n is 1, 2, 3 or 4.

The term "alkyl" refers, in the present invention, to saturated, linear or branched hydrocarbon chains having 1 to 6 carbon atoms, for example, methyl, ethyl, n-propyl, / -propyl, n-butyl , ferc-butyl, sec-butyl, n-pentyl, n-hexyl, etc. Preferably and particularly for some radicals the alkyl group has between 1 and 4 carbon atoms, for example, without being limited, to methyl, ethyl, n-propyl, / -propyl, n-butyl, ferc-butyl, isobutyl or sec-butyl The alkyl groups may be optionally substituted by one or more substituents such as alkynyl, alkenyl, halogen, hydroxyl, alkoxy, carboxyl, cyano, carbonyl, acyl, alkoxycarbonyl, carbamoyl, amino or nitro.

The term "aryl" refers, in the present invention, to aromatic rings, single or multiple, having between 6 and 18 carbon atoms in the ring part, such as but not limited to, phenyl, naphthyl, diphenyl, indenyl, phenanthryl, fluorenyl or anthracil Preferably the aryl group has 6 to 12 carbon atoms and more preferably the aryl group is a phenyl. The aryl radicals can be

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optionally substituted in any of their positions by one or more substituents or two substituents forming an aryl fused cycle and are independently selected from among such as alkyl, alkenyl, alkynyl, O-alkyl, O-aryl, halogen, hydroxyl, amino, substituted amino , carboxylic acid, carboxylic ester or carboxamide.

The term "heteroaryl" refers to an aryl, which contains between 5 and 12 carbon atoms and at least one atom other than carbon, such as S, N, or O, forming part of the aromatic ring. For example, not limited to pyridine, pyrazine, pyrimidine or pyridazine. The heteroaryl radicals may be optionally substituted in any of their positions by one or more substituents or two substituents forming an aryl fused cycle and are independently selected from such as alkyl, alkenyl, alkynyl, O-alkyl, O-aryl, halogen, hydroxy , amino, substituted amino, carboxylic acid, carboxylic ester or carboxamide.

By "aralkyl" is meant in the present invention an aryl group, as defined above, linked to the rest of the molecule by a C1-C4 alkyl group, as defined above. A non-limiting example of aralkyl is a benzyl or phenethyl group. The aralkyl radicals may be optionally substituted in any of their positions by one or more substituents or two substituents forming an aryl fused cycle and are independently selected from among such as alkyl, alkenyl, alkynyl, O-alkyl, O-aryl, halogen, hydroxyl , amino, substituted amino, carboxylic acid, carboxylic ester or carboxamide

The term "heteroaralkyl" refers, in the present invention, to a heteroaryl group, as defined above, linked to the rest of the molecule by a C1-C4 alkyl group, as defined above. The heteroaralkyl radicals can be optionally substituted in any of their positions by one or more substituents or two substituents forming an aryl fused cycle and are independently selected from such as alkyl, alkenyl, alkynyl, O-alkyl, O-aryl, halogen, hydroxyl , amino, substituted amino, carboxylic acid, carboxylic ester or carboxamide.

The stereoisomers of the compounds of the present invention, represented by the formula (I) described above, may include racemic mixture, pure enantiomers, diastereoisomeric mixtures, pure diastereoisomers, depending on the

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presence of one or more stereogenic elements, or isomers, depending on the presence of multiple links (eg, Z, E). Individual isomers, enantiomers or diastereomers and mixtures thereof are included within the scope of the application of the present invention.

In a preferred embodiment, R1 is (C1-C4) alkyl or (C1-C4) aralkyl, more preferably R1 is (C1-C2) alkyl, benzyl or phenethyl, even more preferably R1 is methyl or benzyl.

In another preferred embodiment, when R2 is -OR6, R6 is hydrogen, (C1-C4) alkyl or benzyl, more preferably hydrogen, methyl, ferc-butyl or benzyl.

In another preferred embodiment, when R2 is -NR7R8; R8 is selected from hydrogen, (C1-C6) alkyl, aralkyl (C1-C4), heteroaryl, heteroaralkyl (C1-C4) and -CH (R13) COOR12 and / or R7 is selected from hydrogen and (C1-C4 alkyl) ), more preferably R7 is hydrogen or methyl and / or R8 is selected from benzyl, heteroaryl, heteroaralkyl (C1) and -CH (R13) COOR12; R13 is preferably hydrogen, methyl or benzyl and / or R12 is preferably hydrogen or methyl.

In another preferred embodiment, R3 is hydrogen or methyl.

In another preferred embodiment, R4 is aralkyl (C1-C4) or -COR9. More preferably, R4 is benzyl, -COOR14o -CONR7R8.

When R4 is -COOR14, preferably R14 is hydrogen, (C1-C4) alkyl or benzyl, more preferably hydrogen, methyl, ferc-butyl or benzyl.

When R4 is -CONR7R8; preferably R7 is selected from hydrogen and (C1-C4) alkyl, more preferably hydrogen or methyl and even more preferably hydrogen, and / or R8 is selected from hydrogen, (C1-C6) alkyl and aralkyl (C1-C4), heteroaryl, heteroaralkyl (C1-C4) and -CH (R13) COOR12, more preferably benzyl or -CH (R13) COOCH3, where R13 is preferably selected from benzyl, methyl, heteroaralkyl (C1) or heteroaryl, and even more preferably from benzyl , methyl, pyridinmethyl or pyridine.

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In a preferred embodiment, R5 is -NR11R10 and R11 are selected from hydrogen, methyl and benzyl; and / or R10 is selected from hydrogen, methyl, benzyl and COOR14; where R14 is methyl, tert-butyl or benzyl.

In another preferred embodiment the compound of the invention is selected from the list consisting of:

4R, S-Benzyl-4-methoxycarbonyl-1 - [(3’S-tert-butoxycarbonylamino-3’-

benzyloxycarbonyl) prop-1’-il] -2-oxoazetidine

4R, S-Benzyl-4-methoxycarbonyl-1 - [(3’S-tert-butoxycarbonylamino-3’-methyloxycarbonyl) prop-

1’-il] -2-oxoazetidine

4R, S-Benzyl-4-benzyloxycarbonyl-1 - [(3’S-tert-butoxycarbonylamino-3’-

benzyloxycarbonyl) prop-1’-il] -2-oxoazetidine

4S-Benzyl-4-benzyloxycarbonyl-1 - [(3’S-tert-butoxycarbonylamino-3’-

benzyloxycarbonyl) prop-1’-il] -2-oxoazetidine

4R-Benzyl-4-benzyloxycarbonyl-1 - [(3’S-tert-butoxycarbonylamino-3’-

benzyloxycarbonyl) prop-1’-il] -2-oxoazetidine

4R, S-Benzyl-4-benzyloxycarbonyl-1 - [(3’S-tert-butoxycarbonylamino-3’-

benzyloxycarbonyl) prop-1’-yl] - (3R, S) -methyl-2-oxoazetidine

4R, S-Methyl-4-methoxycarbonyl-1 - [(3’S-tert-butoxycarbonylamino-3’-benzyloxycarbonyl) prop-

1’-il] -2-oxoazetidine

4R, S-Benzyl-4-methoxycarbonyl-1 - [(3’S-benzyloxycarbonylamino-3’-tert-

butoxycarbonyl) prop-1’-il] -2-oxoazetidine

4R, S-Methyl-4-methoxycarbonyl-1 - [(3’S-benzyloxycarbonylamino-3’-tert-butoxycarbonyl) prop-

1’-il] -2-oxoazetidine

4R, S-Benzyl-1- [4’S- (W-Benzyloxycarbonyl-W-methyl) amino-4’-methoxycarbonyl] but-1-yl] -4-

tert-butoxycarbonyl-2-oxoazetidine

4R, S-Benzyl-1- [4’S - [(W-methyl) amino-4’-methoxycarbonyl] but-1’-yl] -4-tert-butoxycarbonyl-2-

oxoazetidine

4R, S-Benzyl-4-benzyloxycarbonyl-1 - [(4’S-tert-butoxycarbonylamino-4’-

benzyloxycarbonyl) but-1’-yl] -2-oxoazetidine

4R, S-Benzyl-N - [(4’S-benzyloxycarbonylamino-4’-methoxycarbonyl) but-1’-il] -4-terc-

butoxycarbonyl-2-oxoazetidine

4’S-N - [(4’-Benzyloxycarbonylamino-4’-methoxycarbonyl) but-1’-il] -4R, S-tert-butyloxycarbonyl-

4-methyl-2-oxoazetidine

4R, S-Benzyl-4-benzyloxycarbonyl-1 - [(2’S-benzyloxycarbonylamino-3’-phenyl) prop-1’-il] -2-

oxoazetidine

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4R, S-Bendl-4-bendloxicarbonil-1 - [(2’R-bendloxicarbonylamino-3’-phenyl) prop-1’-il] -2-

oxoazetidine

4S-Bendl-4-bendloxicarbonil-3S-methyl-1 - [(2’S-bendloxicarbonylamino-3’-phenyl) prop-1’-

il] -2-oxoazetidine

4R-Bendl-4-bendloxycarbonyl-3R-methyl-1 - [(2’S-bendloxicarbonylamino-3’-phenyl] prop-1’-

il] -2-oxoazetidine

4R-Bendl-4-bendloxycarbonyl-3R-methyl-1 - [(2’R-bendloxicarbonylamino-3’-phenyl) prop-1’-

il] -2-oxoazetidine

4S-4-Bendl-4-bendloxicarbonil-3S-methyl-1 - [(2’R-bendloxicarbonylamino-3’-phenyl) prop-

1’-il] -2-oxoazetidine

4S-Bendloxicarbonyl-3S, 4S-dimethyl-1 - [(2’R-bendloxicarbonylamino-3’-phenyl) prop-1’-il] -2-

oxoazetidine

4R, S-Bendl-4-bendloxicarbonil-1 - [(2’S-dibendlamino-3’-phenyl) prop-1’-il] -2-oxoazetidine

4S-Bendl -] - (4-bendloxicarbonil-3S-methyl-1 - [(2’S-dibendlamino-3’-phenyl) prop-1’-il] -2-

oxoazetidine

4R-Bendl-4-bendloxicarbonil-3R-methyl-1 - [(2’S-dibendlamino-3’-phenyl) prop-1’-il] -2-

oxoazetidine

4R, S-Bendl-4-methoxycarbonyl-1 - [(2’S-dibendlamino-3’-phenyl) prop-1’-il] -2-oxoazetidine

4R, S-Bendl-4-tert-butoxycarbonyl-1 - [(2’S-dibendlamino-3-phenyl) prop-1’-il] -2-

oxoazetidine

4S-Bendl-3S-methyl-4-methoxycarbonyl-1 - [(2’S-dibendlamino-3’-phenyl) prop-1’-il] -2-

oxoazetidine

4S-Bendl-4-tert-butoxycarbonyl-3S-methyl-1 - [(2’S-dibendlamino-3’-phenyl) prop-1’-il] -2-

oxoazetidine

4S-Bendl-4-tert-butoxycarbonyl-3S-methyl-1 - [(2’S-bendlamino-3’-phenyl) prop-1’-il] -2-

oxoazetidine

4S-Bendl-4-tert-butoxycarbonyl-3S-methyl-1 - [(2’S-amino-3’-phenyl) prop-1-yl] -2-oxoazetidine

4S-Bendl-4-tert-butoxycarbonyl-3S-methyl-1- [2’S-bendloxycarbonylamino-3’-phenyl) prop-1’-

il] -2-oxoazetidine

4S-Bendl-4-tert-butoxycarbonyl-3S-metiM - [(2’S-tert-butoxycarbonylamino-3-phenyl) prop-

1’-il] -4-oxoazetidine

4R, S-Bendl-4-carboxy-1- [3’S-tert-butoxycarbonylamino-3’-carboxy) prop-1’-il] -2-

oxoazetidine

4R, S-Bendl-1- [4’S-tert-butoxycarbonylamino-4’-carboxy) but-1’-il] -4-carboxy-2-

oxoazetidine

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4S-Bendl-4 - [(N-bendl) carbamoil] -1 - [(3’S-tert-butoxycarbonylamino-3 ’- [(N-

benzyl) carbamoyl] prop-1’-il] -2-oxoazetidine

4R, S-Bendl-1- [4’S-tert-butoxycarbonylamino-4 ’- [(N-bendl) carbamoil] but-1’-il] -4 - [(N-

benzyl) carbamoyl] -2-oxoazetidine

4S-4-Bendl-4 - [(N-bendl) carbamoil] -3S-methyl-1- [3’S-tert-butoxycarbonylamino-3 ’- [(N-

benzyl) carbamoyl] prop-1’-il] -2-oxoazetidine

4R-4-Bendl-4 - [(N-bendl) carbamoyl] -3R-methyl-1- [3’S-tert-butoxycarbonylamino-3 ’- [(N-

benzyl) carbamoyl] prop-1’-il] -2-oxoazetidine

4S-Bendl-4- [N - [(pyridin-4 ’’ - il) methyl] carbamoil-1 - [(3’S-tert-butoxycarbonylamino-3 ’- [N-

[(pyridin-4 ’’ - il) methyl] carbamoil] prop-1’-il] -2-oxoazetidine

4R-Bendl-4- [N - [(pyridin-4 ’’ - il) methyl] carbamoil] -1- [3’S-tert-butoxycarbonylamino-3 ’- [N-

[(pyridin-4 ’’ - il) methyl] carbamoil] prop-1’-il] -2-oxoazetidine

4R, S-Bendl-4 - [(N-pyridin-3-yl) carbamoyl] -1 - [(3’S-tert-butoxycarbonylamino-3 ’- [(N-pyridin-

3-yl) carbamoyl] prop-1’-il] -2-oxoazetidine

4S-Bendl-4 - [(N-pyridin-4-yl) carbamoyl] -1- [3’S-tert-butoxycarbonylamino-3 ’- [(N-pyridin-4-

il) carbamoyl] prop-1’-il] -2-oxoazetidine

4R-Bendl-4 - [(N- (pyridin-4-yl) carbamoyl] -1- [3’S-tert-butoxycarbonylamino-3 ’- [(N-pyridin-4-

il) carbamoyl] prop-1’-il] -2-oxoazetidine

4S-Bendl-1- [3’S-tert-butoxycarbonylamino-3 ’- [N - [(1’’S-

methoxycarbonyl) ethyl] carbamoyl] prop-1’-il] -4- [N - [(1 ’’ ’S-methoxycarbonyl) ethyl] carbamoyl] -2-

oxoazetidine

4R-Bendl-1- [3’S-tert-butoxycarbonylamino-3 ’- [N - [(1’’S-

methoxycarbonyl) ethyl] carbamoyl] prop-1’-il] -4- [N - [(1 ’’ ’S-methoxycarbonyl) ethyl] carbamoyl] -2-

oxoazetidine

4R-Bencil-1- [3’S-tert-butoxycarbonylamino-3 ’- [N - [(1’’S-methoxycarbonyl-2’-

phenyl) ethyl] carbamoyl] prop-1’-il] -4- [N - [(1 ’’ S-methoxycarbonyl-2’phenyl) ethyl] carbamoyl] -2-

oxoazetidine

4S-Bencil-1- [3’S-tert-butoxycarbonylamino-3 ’- [N - [(1’’S-methoxycarbonyl-2’-

phenyl) ethyl] carbamoyl] prop-1’-il] -4- [N - [(1 ’’ S-methoxycarbonyl-2’phenyl) ethyl] carbamoyl] -2-

oxoazetidine

4R-Benzyl-1- [4’S - [(N-benzyloxycarbonyl-N-methyl) amino-4’-methoxycarbonyl] but-1’-yl] -4- [N-

[(1’’S-methoxycarbonyl) ethyl] carbamoyl] -2-oxoazetidine

4R, S-Benzyl-1 - [(4’S-methylamino-4’-methoxycarbonyl) but-1’-il] -4- [N - [(1’’S-

methoxycarbonyl) ethyl] carbamoyl] -2-oxoazetidine

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4R-Benzyl-1 - [(4’S-benzyloxycarbonylamino-4’-methoxycarbonyl) but-1’-yl] -4- [N - [(1’’S-methoxycarbonyl) ethyl] carbamoyl] -2-oxoazetidine, and

4R, S-Benzyl-1 - [(4’S-benzyloxycarbonylamino-4’-methoxycarbonyl) but-1’-il] -4- [N - [(1’’R-

methoxycarbonyl) ethyl] carbamoyl] -2-oxoazetidine.

Another aspect of the present invention relates to the process for obtaining the compound of formula (III), that is to say the compound of formula (I) when R2 is -OR6, which comprises the basic medium cyclization of the compound of formula (II) :

image2

image3

(II)

(III)

Another aspect of the present invention relates to the process for obtaining the compound of formula (IV), that is to say the compound of formula (I) when R2 is -NR7R8, which comprises the reaction of R8R7NH amines with a carboxylic acid derivative of the compound of formula (III), ie the compound of formula (I) when R2 is -OH, in the presence of a coupling agent:

R8

-co2h

or

vl

3

n (L / R4

r7r8nh

image4

R5

R5

(IV)

The coupling agent that can be used is any known to one skilled in the art from among those typically used in the synthesis of peptides such as carbodiimides, phosphonium salts or uronium salts.

The present invention further provides pharmaceutical compositions comprising at least one compound of formula (I), acceptable pharmaceutical salts or

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stereoisomers thereof, together with an acceptable pharmaceutical carrier, adjuvant or vehicle for administration to a patient. Preferably, said composition also comprises another active ingredient with synergistic or complementary effect.

Therefore, another aspect of the present invention relates to pharmaceutical composition comprising at least one compound of general formula (I), in addition to at least one pharmaceutically acceptable vehicle.

Pharmaceutical forms suitable for oral administration include any solid composition (tablets, pills, capsules, granulated forms, etc.) or liquid (solutions, suspensions, emulsions, syrups, etc.) and may contain conventional excipients known in the art, such as binding agents, for example syrup, acacia, gelatin, sorbitol, tragacanth, or polyvinylpyrrolidone; fillers, for example lactose, sugar, starch, corn, calcium phosphate, sorbitol or glycine, lubricants for the preparation of tablets, for example magnesium stearate, disintegrants such as starch, polyvinyl pyrrolidone, sodium starch glycolate or microcrystalline cellulose, or agents pharmaceutically acceptable humectants, such as sodium lauryl sulfate.

Solid oral compositions may be prepared by conventional methods of mixing, filling or preparing tablets. Repeated mixing operations can be used to evenly distribute the active ingredient using large amounts of fillers. These operations are conventional in the art of this invention. The tablets may be prepared, for example, through wet or dry granulation and may optionally be coated by methods well known in normal pharmaceutical practice, particularly with an enteric coating.

Pharmaceutical compositions may also be adapted for parenteral administration, such as sterile solutions, suspensions or lyophilized products in the appropriate pharmaceutical form. Suitable excipients, such as bulk agents, neutralizers or surfactants may be mentioned.

The compounds or compositions described in the present invention can be administered by any suitable method, such as intravenous infusion,

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oral preparations and intraperitoneal or intravenous administration. However, the preferred route of administration will depend on the condition of the patient.

The therapeutically effective amount of the compound of formula (I) to be administered in general will depend, among other factors, on the person to be treated, on the severity of the disease, on the form of administration chosen, etc. For this reason, the doses mentioned in this invention should be considered as guidelines for the specialist in the field, and the latter should adjust the dose according to the variables mentioned above. However, a compound of formula (I) can be administered one or more times a day, for example, 1,2, 3 or 4 times a day in a typical total daily amount between 1 and 200 mg / kg body weightMa, preferably 1-10 mg / kg of body mass / day.

The compounds described in this invention, their pharmaceutically acceptable salts and / or stereoisomers, as well as the pharmaceutical compositions containing them can be used together with other additional drugs to provide a combination therapy. Such additional drugs may be part of the same pharmaceutical composition or, alternatively, be provided in a form of a separate composition for simultaneous or not administration, with the pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable stereoisomer or Get out of it.

Another aspect of the invention relates to the use of the compound of formula (I) for the preparation of a medicament.

Another aspect of the present invention relates to the use of the compound of formula (I) for the preparation of a medicament for the treatment and / or prevention of a disease associated with alterations of the thermosensory channels, preferably of the TRPM8 channels, preferably said diseases are selected from inflammation; pulmonary disorders for example problems of the respiratory system selected between cough and asthma; eye diseases such as dry eye smdrome or excessive tearing; gastrointestinal hypersensitivity such as irritable bowel syndrome; skin sensitivity such as pruritus, atopic dermatitis, allergen or psoriasis; neurodegeneration and cancer such as melanoma, leukemia, prostate, kidney, breast, pancreas, ovary, lung, colon, central nervous system, particularly the brain, or osteosarcoma.

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Throughout the present description, the term "treatment" refers to eliminating, reducing or decreasing the cause or effects of the disease.For the purposes of this invention, treatment includes, but is not limited to, alleviate, decrease or eliminate one or more symptoms of the disease; reduce the degree of disease, stabilize (i.e., not worsen) the disease status, delay or slow the progression of the disease, relieve or improve the disease status and remit the disease disease (either totally or partially).

Throughout the description and the claims the word "comprises" and its variants are not intended to exclude other technical characteristics, additives, components or steps. For those skilled in the art, other objects, advantages and characteristics of the invention will be derived partly from the description and partly from the practice of the invention. The following examples and figures are provided by way of illustration, and are not intended to be limiting of the present invention.

DESCRIPTION OF THE FIGURES

FIG. 1 Response dose curves, representative of the blocking effect of some compounds, of examples 3, 6 and 17 (FIGs. 1A, 1B and 1C, respectively). The percentage of activity of the TRPM8 channel is represented in the presence of increasing concentrations of the compounds tested. The activity is normalized against the activity of the channel in the absence of compound. Solid lines show the adjustment to the Hill equation where the IC50 values are obtained. The results represent the mean ± SD for n> 6 measurements.

EXAMPLES

SYNTHESIS OF THE COMPOUNDS OF THE INVENTION

The compounds of the present invention, derived from p-lactams, have been prepared following the general scheme of synthesis 1.

TO

H2N fsj co2r °

H-Xaa-OR6

ho2c.

On

r5 ^ r4

n = 0,1,2

image5

PhSH,

k2co3

CH, CN

R1

X 6

HN? SfC02R6

l

On

R ^ R4

CICOCH (R3) CI propylene oxide THF

p3 R1 O

R '' <

A — n hn-r

° 3

r6nh2

, 3 R1

-co2h

. N

O] agent n () ^ R4 coupling n () R4

T * T *

R5 R5

EXAMPLES 35-51

EXAMPLES 33-34

, 3 R '

-co2r6

° '3

n (LR4

T *

B:

CH, CN

Or R1

clvY ^ 'N'fsTco2R6

R3

On

r5 ^ r4

6.7

EXAMPLES 1-32

Scheme 1. General procedure for the preparation of p-lactamic derivatives from amino acids.

5

The Nosyl amino acid derivatives (2) are coupled with the alcohols (3), by Mitsunobu reaction, to give rise to the N-nosyl-N-alkyl derivatives (4). Elimination of the nosyl group gives rise to the amino derivatives (5), which are reacted with chloroalkanoyl chlorides to give rise to the intermediates (6). The basic medium cyclization of the intermediates (6) gives rise to the corresponding p-lactam compounds (EXAMPLES 1-32), which are subsequently transformed into the amide analogs (EXAMPLES 35-51) by coupling of the R8NH2 amines to the precursor carboxylates (among others, EXAMPLES 33-34).

15 SYNTHESIS OF STARTING PRODUCTS AND REACTION INTERMEDIATES

Nosil amino acid derivative synthesis (Ns-L-Xaa-OR6) (2)

To a solution of H-L-Xaa-OR6HCl (17.15 mmol) in dry CH2Cl2 (115 mL), triethylamine (TEA) (17.15 mmol, 2.4 mL) is added and allowed to stir for 20 min. After this time, TEA (22.25 mmol, 3.1 mL) and 2-nitrobenzenesulfonyl chloride (22.25 mmol, 4.9 g) are added at 0 ° C, leaving under stirring at room temperature for one night. The next day the solvent is evaporated to dryness and the reaction crude is extracted with AcOEt and washed with dric acid (10%), NaHCO3 (10%) and saturated NaCl solution, successively. Finally the

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Organic residue is dried over anhydrous Na2SO4, filtered and evaporated to dryness. The reaction crude is purified on a silica gel column using the eluent system indicated in each case.

Ns-L-Phe-OMe (2a) (Albanese, D .; Lardini, D .; Lupi, V .; Penso, M. Eur. J. Org. Chem.

2000, 1443-1449)

Ns-L-Phe-O * Bu (2b) (Turner, JJ; Wilschut, N .; Overkleft, HS; Klaffke, W .; Van der Marel, GA; Van Boom, JH Tetrahedron Lett. 1999, 40, 7039- 7042)

Ns-L-Phe-OBn (2c)

image6

Sirupe Rdto: 89%. Eluent: AcOEt: Hexane (3: 1). HPLC: t R = 15.33 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (300 MHz, CDCfe): 5 8.00-6.91 (m, 14H, Ar), 6.07 (d, 1H, J = 9.0 Hz, NH), 4.92 (d, 1H, J = 12.1 Hz, OCH2), 4.87 (d, 1H, J = 12.1 Hz, OCH2), 4.52 (m, 1H, a-Phe), 3.13 (m, 2H, p-Phe). 13C NMR (CDCl3): 5 171.5 (COO), 147.2, 136.6, 136.1, 135.1, 134.4, 132.8, 128.9, 128.6, 128.2, 127.7, 127.6, 127.1,

125.9, 124.2 (Ar), 66.4 (OCH2), 58.7 (Ca), 35.9 (CP). MS (ES) +: 441.21 [M + H] +. Ns-L-Ala-OMe (2d) (Biron, E .; Kessler, H. J. Org. Chem. 2005, 70, 5183-5189) Ns-L-Ala-OBn (2e)

image7

no2

Sirupe Rdto: 62%. Eluent: AcOEt: Hexane (2: 1). HPLC: t R = 13.04 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (400 MHz, CDCh): 5 8.00 (dd, 1H, J = 7.7, 1.6 Hz, Ar), 7.81 (dd, 1H, J = 7.7, 1.6 Hz, Ar), 7.34-7.31 (m, 3H, Ar), 7.20-7.17 (m, 2H, Ar), 6.18 (d, 1H, J = 8.6 Hz, NH), 4.96 (d, 1H, J = 12.2 Hz, OCH2), 4.91 (d, 1H, J = 12.1 Hz, OCH2), 4.31 (m, 1H, aH), 1.50 (d, 3H, J = 7.2 Hz, CH3). 13C NMR (75 MHz, CDCh): 171.4 (COO), 147.6, 134.8, 134.1, 133.7, 132.9, 130.4, 128.7, 128.6, 128.3, 125.7 (C, Ar), 67.4 (OCH2), 52.6 (Ca), 19.8 (CH3). MS (ES) +: 365.09 [M + H] +.

Ns-L-Ala-OfBu (2f) (Chapman R. N; Dimartino, G .; Arora, P. S. J. Am. Chem. Soc.

2004, 126, 12252-12253)

Reduction of amino acid derivatives to the corresponding alcohols (3)

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A solution of the corresponding amino acid derivative (12.38 mmol) in dry THF (12 mL) is added successively, at -15 ° C, W-methylmorpholine (12.38 mmol, 1.36 mL) and the chloroformate of isobutyl (12.38 mmol, 1.60 mL). After 10 min the precipitate of W-methylmorpholinium hydrochloride formed is removed by filtration, being washed with THF (2x60 mL). The solution thus obtained is cooled to -15 ° C and then a solution of NaBH4 (18.57 mmol, 0.7 g) in H2O (6 mL) is added.

After the release of H2, H2O (300 mL) and AcOEt (600 mL) are added, separating the two phases. The organic phase is washed successively with dric acid (10%), NaHCO3 (10%) and saturated NaCl solution, successively. Finally, the organic residue is dried over anhydrous Na2SO4, filtered and evaporated to dryness. The reaction crude is purified on a silica gel column using the eluent system indicated in each case.

Boc-L-Hse-OBn (3a) (commercial)

Z-L-Hse-O * Bu (3b) (commercial)

(2S) -2- (N-Methyl benzyloxycarbonyl-N-methyl) amino-5-hydroxypentanoate (3c)

image8

Sirupe Rdto: 58%. Eluent: AcOEt: Hexane (1: 1). Proportion of rotamers M, m = 2: 1. 1H NMR (300 MHz, CDCh, majority rotamer): 5 7.40 (s, 5H, Ph), 5.20 (s, 2H, OCH2), 4.90 (dd, 1H, J = 10.5, 4.9 Hz, 2-H), 4.41 (m, 2H, 5-H), 3.75 (s, 3H, OMe), 2.92 (s, 3H, NMe), 2.08 (m, 2H, 3-H), 1.85 (m, 1H, 4-H), 1.62 (m, 1H, 4-H). MS (ES) +: 318.14 [M + Na] +.

(2S) -2-ferc-Butoxycarbonylamino-5-benzyl hydroxypentanoate (3d) (Jiang, S .; Li, P .; Lai, CC; Kelley, JA; Roller, PPJ Org. Chem. 2006, 71, 7307- 7314.)

(2S) -2- Methyl benzyloxycarbonylamino-5-hydroxypentanoate (3e) (Feichtinger, K .; Sings, H. L .; Baker, T.J.; Mathews, K .; Goodman, M. J. Org. Chem. 1998, 63, 84328439)

Z-S-phenylalaninol (3f) (commercial)

Z-R-phenylalaninol (3g) (commercial)

(2S) -dibenzylaminophenylalaninol (3h) (commercial)

Synthesis of M-alkyl-M-nosyl amino acid derivatives

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At a solution of the corresponding derivative alcohol (7.60 mmol), Ns-L-Xaa-OR6 (7.60 mmol) and triphenylphosphine (7.60 mmol, 2g) in dry THF (66 mL), under Ar atmosphere At 0 ° C, diisopropyl azodicarboxylate (7.60 mmol, 1.50 mL) is added. The ice bath is removed and left under stirring at room temperature overnight. The next day the solvent is evaporated to dryness and the reaction crude is purified on a silica gel column using the eluent system indicated in each case.

W - [(3S-ferc-Butoxicarbomlammo-3-bencMoxicarboml) prop-1-M] -Ns-L-Phe-OMe

(4th)

image9

Sirupe Rdto: 50% [from Ns-L-Phe-OMe (2a) and Boc-L-Hse-OBn (3a)]. Eluent: AcOEt: Hexane (1: 2). HPLC: t R = 16.80 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (300 MHz, CDCh): 5 7.76- 7.18 (m, 14H, Ar), 5.20 (d, 1H, J = 12.2 Hz, OCH2), 5.16 (d, 1H, J = 12.2 Hz, OCH2), 5.15 (brs, 1H, NH Boc), 4.93 (t, 1H, J = 7.7 Hz, a-Phe), 4.26 (m, 1H. 3-H), 3.54 (m, 1H, 1-H), 3.53 (s , 3H, OMe), 3.40 (m, 1H, 1-H), 3.33 (dd, 1H, J = 14.5, 7.1 Hz, P-Phe), 2.92 (dd, 1H, J = 14.5, 8.4 Hz, P- Phe), 2.17 (m, 1H, 2-H), 1.99 (m, 1H, 2-H), 1.45 (s, 9H, CH3 fBu). 13C NMR (75 MHz, CDCl3): 171.8 (COO), 170.9 (COO), 155.6 (CON), 148.3, 136.0, 135.3, 133.7, 132.9, 131.7, 131.0, 129.1,

128.9, 128.8, 128.7, 128.6, 127.2, 124.2 (C, Ar), 80.3 (C, * Bu), 67.6 (OCH2), 61.5 (Ca-Phe), 52.5 (OMe), 52.00 (C3), 43.0 (C1 ), 36.6 (P-Phe), 33.3 (C2), 28.5 (CH3 fBu). MS (ES) +: 656.38 [M + H] +.

W - [(3S-ferc-Butoxycarbonylamino-3-benzyloxycarbonyl) prop-1-yl] -Ns-L-Phe-OBn

(4b)

image10

Sirupe Rdto: 38% [from Ns-L-Phe-OMe (2c) and Boc-L-Hse-OBn (3a)]. Eluent: AcOEt: Hexane (1: 3). HPLC: t R = 18.23 min (gradient from 5% to 100% of A, in 20 min).

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1H NMR (400 MHz, CDCI3): 5 7.72-7.12 (m, 19H, Ar), 5.17 (s, 2H, OCH2), 5.12 (d, 1H, J = 7.7 Hz, NH, Boc), 4.98 (d, 1H, J = 13.8 Hz, OCH2), 4.91 (d, 1H, J = 13.8 Hz, OCH2), 4.90 (m, 1H, a-Phe), 4.25 (m, 1H, 3-H), 3.55 (m, 1H, 1-H), 3.37 (m, 2H, p-Phe, 1-H), 2.94 (dd, 1H, J = 14.8, 8.0 Hz, p-Phe), 2.16 (m, 1H, 2-H) , 1.96 (m, 1H, 2-H), 1.45 (s, 9H, CH3 fBu). 13C NMR (75 MHz, CDCl3): 171.7 (COO), 170.1 (COO), 155.5 (CON), 148.1,

135.9, 135.3, 134.9, 133.6, 132.7, 131.5, 130.8, 129.2, 128.9, 128.85, 128.8, 128.7,

128.6, 128.5, 128.5, 127.1, 124.1 (C, Ar), 80.2 (C, fBu), 67.52 (OCH2), 67.5 (OCH2),

61.5 (Ca-Phe), 52.00 (C3), 43.0 (C1), 36.8 (Cp-Phe), 33.2 (C2), 28.4 (CH3 fBu). MS (ES) +: 732.33 [M + H] +.

W - [(3S-ferc-Butoxicarbomlammo-3-bencMoxicarboml) prop-1-M] -Ns-L-Ala-OMe

(4c)

no2

C02Me C02Bn

Sirupe Rdto: 6% [from Ns-L-Ala-OMe (2d) and Boc-L-Hse-OBn (3a)]. Eluent: AcOEt: Hexane (1: 2). HPLC: t R = 17.03 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (400 MHz, CDCI3): 5 7.96 (d, 1H, J = 7.8 Hz, Ns), 7.86-7.56 (m, 3H, Ns), 7.36 (s, 5H, Ph), 5.21 (d, 1H, J = 12.3 Hz, OCH2), 5.18 (sa, 1H, NHBoc), 5.16 (d, 1H, J =

12.3 Hz, OCH2), 4.76 (m, 1H, a-Ala), 4.27 (m, 1H, 3-H), 3.57 (s, 3H, OMe), 3.51 (m, 1H, 1-H), 3.22 ( m, 1H, 1-H), 2.23 (m, 1H, 2-H), 2.09 (m, 1H, 2-H), 1.47 (d, 3H, J = 7.4 Hz, CH3, Ala), 1.43 (s , 9H, CH3 fBu). 13C NMR (100 MHz, CDCI3): 171.8 (COO), 165.2 (COO), 155.5 (CON), 148.1, 135.3, 133.7, 131.7, 131.1, 128.8, 128.7, 128.6, 128.5, 124.2 (Ar), 80.3 (C fBu), 67.6 (OCH2), 56.2 (Ca-Ala), 52.5 (OMe), 51.9 (C3), 42.7 (C1), 33.7 (C2), 28.4 (CH3 fBu), 16.8 (CH3 Ala). MS (ES) +: 602.41 [M + Na] +. W - [(3S-Benzyloxycarbonylamino-3-ferc-butoxycarbonyl) prop-1-yl] -Ns-L-Phe-OMe

/: - <n

image11

(4d)

image12

Sirupe Rdto: 52% [from Ns-L-Phe-OMe (2a) and Z-L-Hse-OtBu (3b)]. Eluent: AcOEt: Hexane (1: 2). HPLC: t R = 16.97 min (gradient from 5% to 100% of A, in 20 min).

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1H NMR (400 MHz, CDCI3): 5 7.77 (d, 1H, J = 7.8 Hz, Ar), 7.66- 7.49 (m, 3H, Ar), 7.397.21 (m, 10H, Ph, Z), 5.43 ( d, 1H, J = 7.7 Hz, NH, Z), 5.17 (d, 1H, J = 12.3 Hz, OCH2), 5.10 (d, 1H, J = 12.3 Hz, OCH2), 4.93 (t, 1H, J = 7.7 Hz, a-Phe), 4.18 (m, 1H, 3-H), 3.54 (s, 3H, OMe), 3.51 (dd, 1H, J = 11.8, 3.8 Hz, 1-H), 3.42 (dd, 1H, J = 11.8, 5.4 Hz, 1-H), 3.36 (m, 1H, p-Phe), 2.95 (dd, 1H, J = 14.4, 8.4, Hz, P-Phe), 2.15 (m, 1H, 2-H), 1.98 (m, 1H, 2-H), 1.47 (s, 9H, CH3 fBu). 13C NMR (75 MHz, CDCI3): 170.8 (COO), 170.6 (COO), 156.0 (CON), 148.2, 135.9, 133.7, 131.9, 130.9, 129.1, 128.7, 128.6, 128.2, 128.1, 127.1, 124.1 (C, Ar), 82.8 (C, fBu), 67.0 (OCH2), 61.3 (Ca-Phe), 52.8 (C3), 52.5 (OMe), 42.9 (C1), 36.5 (Cp-Phe), 33.4 (C2), 28.0 (CH3 fBu). MS (ES) +: 678.51

[M + Na] +.

W - [(3S-BencMoxicarbomlammo-3-ferc-butoxycarboml) prop-1-M] -Ns-L-Ala-OMe

(4e)

image13

Sirupe Rdto: 58% [from Ns-L-Ala-OMe (2d) and Z-L-Hse-O * Bu (3b)]. Eluent: AcOEt: Hexane (1: 2). HPLC: t R = 15.49 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (400 MHz, CDCI3): 5 8.00 (dd, 1H, J = 7.9, 1.4 Hz, Ar-Ns), 7.67 (dd, 1H, J =

7.6, 1.4 Hz, Ar-Ns), 7.63-7.58 (m, 2H, Ar-Ns), 7.37-7.31 (m, 5H, Ph, Z), 5.40 (d, 1H, J = 7.1 Hz, NH, Z ), 5.15 (d, 1H, J = 12.0 Hz, OCH2), 5.09 (d, 1H, J = 12.0 Hz, OCH2), 4.76 (q, 1H, J = 7.4 Hz, a-Ala), 4.19 (m, 1H, 3-H), 3.58 (s, 3H, OMe), 3.48 (m, 1H, 1-H), 3.23 (m, 1H, 1-H), 2.14 (m, 2H, 2-H), 1.47 (m, 12H, CH3 * Bu, CH3 Ala). 13C NMR (75 MHz, CDCI3): 171.7 (COO), 170.6 (COO), 156.0 (CON), 148.0, 136.4, 133.7, 131.7, 131.0,

128.6, 128.3, 128.1, 124.2 (C, Ar), 82.8 (C, fBu), 67.0 (OCH2), 56.1 (Ca-Ala), 52.8 (C3), 52.5 (OMe), 42.7 (C1), 33.8 (C2 ), 28.0 (CH3 fBu), 16.8 (CH3 Ala). MS (ES) +: 602.48 [M + Na] +.

W- [4S - [(W-Benzyloxycarboml-N-metM) ammo-4-methoxycarboml] but-1-M] -Ns-L-Phe- O‘Bu (4f)

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Sirupe Rdto: 91% [from Ns-L-Phe-O * Bu (2b) and (3c)]. Eluent: AcOEt: Hexane (1: 1). HPLC: t R = 17.61 min (gradient from 5% to 100% of A, in 20 min). Proportion of rotamers M, m = 2: 1. 1H NMR (400 MHz, CDCl3, majority rotamer): 5.87 (dd, 1H, J = 7.8, 1.2 Ar-Ns), 7.60 (m, 1H, Ar-Ns), 7.53 (m, 2H, Ar-Ns) , 7.26 (m, 10H, Ph, Z), 5.16 (d, 1H, J = 12.5 Hz, OCH2), 5.12 (d, 1H, J = 12.5 Hz, OCH2), 4.79 (t, 1H, J = 7.8 Hz , a-Phe), 4.74 (dd, 1H, J = 10.5, 4.8, 4-H), 3.69 (s, 3H, OMe), 3.55 (m, 1H, 1-H), 3.31 (dd, 1H, J = 14.3, 7.9 Hz, P-Phe), 3.24 (m, 1H, 1-H), 2.97 (dd, 1H, J = 14.3, 7.9 Hz, P-Phe), 2.83 (s, 3H, NMe), 1.92 (m, 1H, 3-H), 1.71 (m, 3H, PH, 2-H), 1.21 (s, 9H, CH3 fBu). 13C NMR (75 MHz, CDCI3): 171.8 (COO), 169.2 (COO), 157.0 (CON), 148.3,

136.7, 133.5, 131.7, 129.2, 128.7, 128.6, 127.9, 126.9, 123.9 (C, Ar), 82.4 (C, fBu),

67.6 (OCH2), 61.8 (Cp-Phe), 57.9 (C4), 52.3 (OMe), 45.8 (C1), 36.9 (Cp-Phe), 30.6 (NMe), 27.7 (C ^ 'Bu), 27.1 (C3 ), 26.1 (C2). MS (ES) +: 706.68 [M + Na] +. W - [(4S-ferc-Butoxicarbomlammo-4-benzyloxycarboml) but-1-M] -Ns-L-Phe-OBn (4g)

image15

Sirupe Rdto: 62% [from Ns-L-Phe-OBn (2c) and 3d]. Eluent: AcOEt: Hexane (1: 2). tR = 18.60 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (300 MHz, CDCI3): 5 7.87-6.84 (m, 19H, Ar), 5.11 (d, 1H, J = 12.0 Hz, OCH2), 5.06 (d, 1H, J = 12.0 Hz, OCH2), 4.93 (m, 1H, NH), 4.90 (d, 1H, J = 12.1 Hz, OCH2), 4.85 (d, 1H, J = 12.2 Hz, OCH2), 4.81 (m, 1H, a-Phe), 4.21 (m , 1H, 4-H), 3.40 (ddd, 1H, J = 15.3, 9.9, 5.2 Hz, 1- H), 3.27 (dd, 1H, J = 14.1, 8.0 Hz, P-Phe), 3.17 (m, 1H, 1-H), 2.92 (dd, 1H, J = 14.0, 7.1 Hz, P-Phe), 1.60 (m, 4H, 3-H, 2-H), 1.44 (s, 9H, CH3 * Bu) . 13C NMR (CDCI3): 5 172.4, 170.1 (COO), 155.5 (OCON), 148.2, 136.3, 135.4, 134.9, 133.5, 133.0, 131.5, 131.0, 129.3, 128.8, 128.7, 128.6, 128.5, 128.4, 127.1, 124.1 (Ar), 80.1 (C, * Bu), 67.5, 67.3

(OCH2), 61.5 (Ca-Phe), 53.1 (C4), 46.1 (C1), 36.9 (Cp-Phe), 30.1 (C3), 28.4 (C ^ 'Bu),

26.4 (C2). MS (ES) +: 746.29 [M + H] +.

W - [(4S-benzyloxycarbonylamino-4-methyloxycarbonyl) -but-1-yl] -Ns-L-Phe-OlBu (4h)

image16

5 Sirupe. Rdto: 68% [from Ns-L-Phe-O * Bu (2b) and (2S) -2-benzyloxycarbonylamino-5- methyl hydroxypentanoate (3e)]. HPLC-MS: t R = 8.53 min (gradient from 30% to 95% of A, in 10 min). 1H NMR (400 MHz, CDCI3): 5 7.84 (d, 1H, J = 7.8 Hz, 3-H Nos), 7.63 (m, 1H, 6-H Nos), 7.56 (m, 2H, 4.5-H Nos), 7.36-7.20 (m, 10H, Ar), 5.33 (d, 1H, J = 8.2 Hz, 4-NH), 5.11 (s, 2H, OCH2), 4.75 (t, 1H, J = 7.6 Hz, P-Phe), 4.33 (m, 1H, 4-H), 3.74 (s, 10 3H, OMe), 3.55-3.48 (m, 1H, 1-H), 3.35-3.27 (m, 2H, 1-H , P-Phe), 3.99 (dd, 1H, J =

14.5, 7.1 Hz, P-Phe), 1.87-1.80 (m, 1H, 3-H), 1.79-1.58 (m, 3H, 3-H, 2-H), 1.23 (s, 9H, CH3 fBu). MS (ES) +: 670.48 [M + H] +.

W - [(4S-benzyloxycarbonylamino-4-methyloxycarbonyl) -but-1-yl] -Ns-L-Ala-OlBu (4i)

image17

15 Sirupe. Rdto: 63% [from Ns-L-Ala-OMe (2d) and (2S) -2-benzyloxycarbonylamino-5- methyl hydroxypentanoate (3e)]. HPLC-MS: t R = 7.62 min (gradient from 30% to 95% of A, in 10 min). 1H NMR (400 MHz, CDCI3): 5 8.05 (d, 1H, J = 6.5 Hz, 3-H Nos), 7.64 (m, 2H, 4.5-H Nos), 7.57 (m, 1H, 6-H Nos), 7.36-7.30 (m, 5H, Ar), 5.34 (d, 1H, J = 7.9 Hz, 4- NH), 5.11 (s, 2H, OCH2), 4.62 (q, 1H, J = 7.2 Hz, a-Ala), 4.35 (q, 1H, J = 7.2 Hz, 4-H), 20 3.74 (s, 3H, OMe), 3.52-3.44 (m, 1H, 1-H), 3.16-3.09 (m, 1H, 1-H), 1.86-1.64 (m, 4H,

3-H, 2-H), 1.45 (d, 3H, J = 7.3 Hz, p-Ala), 1.32 (s, 9H, CH3 fBu). MS (ES) +: 594.32 [M + H] +.

W - [(2S-Benzyloxycarbomlammo-3-feml) prop-1-M] -Ns-L-Phe-OBn (4j)

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Sirupe Rdto: 51% [from Ns-L-Phe-OBn (2c) and 3f]. Eluent: AcOEt: Hexane (1: 3). HPLC: t R = 18.52 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (400 MHz, CDCI3): 5 7.79-7.02 (m, 24H, Ar), 5.42 (d, 1H, J = 7.4 Hz, 2-NH), 5.10 (d, 1H, J = 12.6 Hz, OCH2) , 5.00 (d, 1H, J = 12.6 Hz, OCH2), 4.84 (s, 2H, OCH2), 4.82 (dd, 1H, J = 8.4, 6.6 Hz, a-Phe), 4.14 (m, 1H, 2- H), 3.55 (dd, 1H, J = 15.5, 10.1 Hz, 1-H), 3.43 (dd, 1H, J = 15.5, 4.9 Hz, 1-H), 3.24 (dd, 1H, J = 13.9, 8.5 Hz, p-Phe), 3.11 (dd, 1H, J = 13.9,

6.6 Hz, P-Phe), 2.89 (dd, 1H, J = 14.0, 6.0 Hz, 3-H), 2.81 (dd, 1H, J = 14.0, 7.2 Hz, 3-H). 13C NMR (75 MHz, CDCI3): 5 170.0 (COO), 156.0 (CON), 147.8, 137.3, 136.1, 134.7, 133.6, 133.0, 131.9, 131.1, 129.5, 129.4, 128.6, 128.7, 128.66, 128.65, 128.63, 128.62, 128.5, 128.1, 127.9, 127.1, 126.8, 124.4 (Ar), 67.5, 66.5 (OCH2), 62.0 (Ca-Phe), 52.2 (C2), 48.7 (C1), 39.6 (Cp-Phe), 37.3 ( C3). MS (ES) +: 708.43 [M + H] +. W - [(2R-Benzyloxycarbomlammo-3-feml) prop-1-M] -Ns-L-Phe-OBn (4k)

image19

Sirupe Rdto: 57% [from Ns-L-Phe-OBn (2c) and 3g]. Eluent: AcOEt: Hexane (1: 3). HPLC: t R = 18.64 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (400 MHz, CDCI3): 5 7.69-6.88 (m, 24H, Ar), 5.22 (d, 1H, J = 7.4 Hz, NH, Z), 5.01 (d, 1H, J = 12.2 Hz, OCH2) , 5.00 (s, 2H, OCH2), 4.97 (d, 1H, J = 12.2 Hz, OCH2), 4.83 (t, 1H, J = 7.6 Hz, a-Phe), 4.06 (m, 1H, 2-H) , 3.64 (dd, 1H, J = 15.8, 9.3 Hz, 1-H), 3.45 (dd, 1H, J = 15.8, 4.5 Hz, 1-H), 3.23 (dd, 1H, J = 14.5, 7.6 Hz, P-Phe), 2.99 (dd, 1H, J = 13.9, 6.9 Hz, 3-H), 2.79 (m, 2H, p-Phe, 3-H). 13C NMR (75 MHz, CDCI3): 5 170.1 (COO), 156.1 (CON), 148.0, 137.8, 135.8, 135.1, 133.7, 132.6, 131.6, 131.1, 129.4, 129.0, 128.75,

128.7, 128.6, 128.5, 128.05, 127.0, 126.8, 124.2 (Ar), 67.7, 66.5 (OCH2), 61.0 (Ca-Phe), 52.4 (C2), 48.4 (C1), 39.2 (C3), 35.6 (Cp- Phe) MS (ES) +: 708.42 [M + H] +. W - [(2R-Benzyloxycarbomlammo-3-feml) prop-1-M] -Ns-L-Ala-OBn (4l)

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image20

Sirupe Rdto: 57% [from Ns-L-Ala-OBn (2e) and 3g]. Eluent: AcOEt: Hexane (1: 3). HPLC: t R = 16.91 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (400 MHz, CDCI3): 5 7.76 (d, 1H, J = 7.9 Hz, Ar), 7.58-7.14 (m, 13H, Ar), 5.15 (d, 1H, J = 7.4 Hz NH, Z), 5.00 (m, 4H, OCH2), 4.69 (q, 1H, J = 7.3 Hz, a-Ala), 3.99 (m, 2H, 2-H), 3.46 (m, 2H,

1-H), 2.97 (dd, 1H, J = 13.8, 6.3 Hz, 3-H), 2.89 (dd, 1H, J = 13.8, 7.4 Hz, 3-H), 1.36 (d, 1H, J = 7.3 Hz, CH3). 13C NMR (75 MHz, CDCI3): 5 171.1 (COO), 156.0 (CON), 148.0,

137.7, 135.2, 133.7, 131.7, 131.2, 129.4, 129.3, 128.7, 128.6, 128.5, 128.45, 128.4, 128.3, 128.1, 128.0, 126.7, 124.2 (Ar), 67.5, 66.6 (OCH2), 56.4 (Ca-Phe) , 52.7 (C2), 48.2 (C1), 38.7 (C3), 16.01 (CH3). MS (ES) +: 632.33 [M + H] +. W- [2S- (3-Feml-2-dibenzylamm) prop-1 -il] -Ns-L-Phe-OBn (4m)

image21

Sirupe Rdto: 63% (from 2c and 3h). Eluent: AcOEt: Hex (1: 5). HPLC: t R = 18.40 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (400 MHz, CDCI3): 5 7,526.77 (m, 29H, Ar), 4.66 (s, 2H, OCH2), 4.54 (dd, 1H, J = 9.7, 5.5 Hz, a-Phe), 3.83 ( dd, 1H, J = 14.8, 5.7 Hz, 1-H), 3.68 (d, 2H, J = 13.7 Hz, N-CH2), 3.53 (d, 2H, J = 13.7 Hz, N-CH2), 3.32 ( dd, 1H, J = 14.8, 8.5 Hz, 1-H), 3.20 (m, 1H, 2-H), 2.81 (dd, 1H, J = 13.9, 8.5 Hz, 3-H), 2.67 (m, 3H , p-Phe, 3-H). 13C NMR (75 MHz, CDCI3): 5 169.7 (COO), 148.0, 139.9, 139.8, 136.4, 135.0, 134.0, 133.3, 131.9, 131.8, 129.7, 129.4, 129.1, 128.6, 128.55, 128.5, 128.4, 128.3, 128.2 , 127.0, 126.9 (Ar), 67.1 (OCH2), 62.0 (Ca-Phe), 57.8 (C2), 53.2 (N-CH2), 47.1 (C1), 36.7 (Cp-Phe), 34.4 (C3). MS (ES) +: 754.35 [M + H] +.

W - [(2S-DibencMammo-3-feml) propyl] -Ns-L-Phe-OMe (4n)

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image22

Sirupe Rdto: 35% (from 2a and 3h). Eluent: AcOEt: Hexane (1: 5). HPLC: t R = 3.60min (gradient of 80% to 95% of A, in 10 min). 1H NMR (300 MHz, CDCl3): 5 7.63 (m, 2H, Ar), 7.41 (m, 1H, Ar), 7.29-7.13 (m, 17H, Ar), 7.02 (m, 2H, Ar), 6.93 ( m, 2H, Ar), 4.54 (dd, 1H, J = 10.0, 5.4 Hz, a-Phe), 3.91 (dd, 1H, J = 15.0, 5.6 Hz, 1-H), 3.80 (dd, 2H, J = 13.7 Hz, NCH2), 3.63 (dd, 2H, J = 13.7 Hz, NCH2), 3.49 (dd, 2H, J =

15.0, 8.7 Hz, 1-H), 3.34 (s, 3H, OCH3), 3.30 (m, 1H, 2-H), 2.89 (dd, 1H, J = 14.0, 8.8 Hz, 3-H), 2.80 ( dd, 1H, J = 13.5, 6.3 Hz, 3-H), 2.75 (dd, 1H, J = 13.2, 10.0 Hz, P-Phe), 2.66 (dd, 1H, J = 13.3, 5.4 Hz, P-Phe ). 13C NMR (75 MHz, CDCI3): 5 170.4 (COO), 140.0-124.5 (C, Ar), 61.8 (a-Phe), 57.8 (C2), 53.3 (NCH2), 52.2 (OCH3), 47.1 (C1) ,

36.6 (P-Phe), 34.7 (C3). MS (ES) +: 679.51 [M + H] +. W - [(2S-Dibenzylamino-3-phenyl) propyl] -Ns-L-Phe-OfBu (4th)

image23

Sirupe Rdto: 82% (from 2b and 3h). Eluent: AcOEt: Hexane (1: 6). HPLC: t R = 5.00 min (80% to 95% gradient of A, in 10 min). 1H NMR (300 MHz, CDCI3): 5 7.59 (m, 2H, Ar), 7.37 (m, 1H, Ar), 7.26-7.16 (m, 16H, Ar), 7.08 (m, 3H, Ar), 6.86 ( m, 2H, Ar), 4.51 (t, 1H, J = 7.7 Hz, a-Phe), 4.01 (dd, 1H, J = 15.0, 5.6 Hz, 1-H), 3.81 (d, 2H, J = 13.7 Hz, NCH2), 3.63 (dd, 2H, J = 13.8 Hz, NCH2), 3.45 (dd, 1H, J = 15.0, 8.8 Hz, 1- H), 3.29 (ddd, 1H, J = 8.6, 5.8, 2.8 Hz, 2-H), 2.83 (m, 2H, 3-H), 2.75 (m, 2H, P-Phe), 1.11 (s, 9H, fBu). 13C NMR (75 MHz, CDCI3): 5 169.0 (COO), 148.2, 140.1, 139.9,

136.8, 134.6, 133.3, 132.0, 129.7, 129.5, 128.4, 121.3, 128.2, 127.0, 126.9, 126.1,

124.4 (C, Ar), 82.0 (C, fBu), 62.6 (a-Phe), 57.9 (C2), 53.3 (NCH2), 47.0 (C1), 37.1 (P-Phe), 34.7 (C3), 27.7 ( CH3, fBu). MS (ES) +: 721.38 [M + H] +.

Elimination of the Nosilo group (Ns)

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To a solution of the corresponding derivative W-Nosyl-alkyl amino acid (4.87 mmol, 3.57 g) in dry CH3CN (82 mL) is successively added K2CO3 (14.49 mmol, 2.02 g) and thiophenol (9 , 74 mmol, 0.99 mL) and stir at room temperature overnight. After this time, the solvent is evaporated to dryness and the resulting residue is dissolved in AcOEt: H2O (1: 1) and the phases are separated. The organic extract is washed with saturated NaCl solution, dried over anhydrous Na2SO4 and evaporated to dryness. The reaction crude obtained is purified on a silica gel column, using the eluent system indicated in each case.

W - [(3S-ferc-Butoxicarbomlammo-3-bencMoxicarboml) prop-1-M] -L-Phe-OMe (5a)

image24

Sirupe Rdto: 85% (from 4th). Eluent: AcOEt: Hexane (1: 1). HPLC: t R = 11.98 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (300 MHz, CDCh): 5 7.38-7.16 (m, 10H, Ar), 5.81 (d, 1H, J = 7.6 Hz, NHBoc), 5.22 (d, 1H, J = 12.5 Hz, OCH2), 5.14 (d, 1H, J = 12.5 Hz, OCH2), 4.41 (m, 1H, 3-H), 3.66 (s, 3H, OMe), 3.50 (t, 1H, J = 6.7 Hz, a- Phe), 2.94 (m, 2H, p-Phe), 2.75 (m, 1H, 1-H), 2.51 (m, 1H, 1-H), 1.96 (m, 1H, 2-H), 1.82 (m, 1H, 2 -H), 1.48 (s, 9H, CH3 fBu) .13C NMR (75 MHz, CDCh): 172.9 (COO),

170.7 (COO), 155.9 (OCON), 136.1, 128.7, 128.5, 127.4, 127.1, 126.9 (C, Ar), 79.9 (C fBu), 65.1 (OCH2), 55.2 (Ca-Phe), 52.5 (OMe), 52.0 (C3), 41.5 (C1), 35.1 (Cp-Phe),

28.6 (C2), 28.3 (CH3 fBu). MS (ES) +: 471.02 [M + H] +.

W - [(3S-ferc-Butoxycarbonylamino-3-benzyloxycarbonyl-) prop-1-yl] -L-Phe-OBn (5b)

image25

Sirupe Rdto: 90% (from 4b). Eluent: AcOEt: Hexane (1: 1). HPLC: t R = 15.47 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (400 MHz, CDCh): 5 7.33-7.07 (m, 15H, Ar), 5.75 (d, 1H, J = 7.7 Hz, NH, Boc), 5.17 (d, 1H, J = 12.4 Hz, OCH2) , 5.10 (d, 1H, J = 12.4 Hz, OCH2), 5.04 (s, 2H, OCH2), 4.37 (m, 1H, 3-H), 3.50 (t, 1H, J = 6.7 Hz, a-Phe) , 2.94 (dd, 1H, J = 13.5, 6.6 Hz, P-Phe), 2.86 (dd, 1H, J = 13.5, D7.3 Hz, P-Phe), 2.72 (m, 1H, 1-H), 2.47 (m, 1H, 1-H), 1.92 (m, 1H, 2-H), 1.79 (m, 1H, 2-H), 1.44 (s, 9H, CH3 fBu). 13C NMR (75 MHz, CDCh): 174.2 (COO), 172.6 (COO), 155.6 (CON), 137.0,

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135.62, 135.6, 129.3, 128.7, 128.6, 128.55, 128.5, 128.4, 126.8 (C, Ar), 79.8 (C, Bu),

67.0, 66.6 (OCH2), 62.7 (a-Phe), 52.5 (C3), 44.2 (C1), 39.8 (P-Phe), 31.7 (C2), 28.5 (CH3 fBu). MS (ES) +: 547.92 [M + H] +.

W - [(3S-ferc-Butoxicarbomlammo-3-bencMoxicarboml) prop-1-M] -L-Ala-OMe (5c)

HN ^ C02Me

C02Bn

Sirupe Rdto: 43% (from 4c). Eluent: AcOEt: Hexane (2: 1). HPLC: t R = 12.15 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (400 MHz, CDCI3): 5 7.34 (s, 5H, Ph), 5.76 (d, 1H, J = 6.0 Hz, 3-NH), 5.18 (d, 1H, J = 16.0 Hz, OCH2), 5.11 (d, 1H, J = 16.0

Hz, OCH2), 4.39 (m, 1H, 3-H), 3.69 (s, 3H, OMe), 3.28 (q, 1H, J = 7.0 Hz, a-Ala), 2.71 (m, 1H, 1-H ), 2.49 (m, 1H, 1-H), 1.87 (m, 2H, 2-H), 1.43 (s, 9H, CH3, Bu,), 1.24 (d, 1H, J = 7.0 Hz, CH3, Ala ). 13C NMR (100 MHz, CDCI3): 175.9 (COO), 172.6 (COO), 155.69 (CON), 135.6, 128.7. 128.4, 128.3, 127.1 (C, Ar), 79.9 (C, Bu), 67.0 (OCH2), 56.7 (Ca-Ala), 52.5 (C3), 51.9 (OMe), 43.9 (C1), 31.9 (C2), 28.4 (CH3 fBu), 19.1 (CH3 Ala). MS (ES) +: 395.35 [M + H] +.

W - [(3S-Benzyloxycarbonylamino-3-ferc-butoxycarbonyl) prop-1-yl] -L-Phe-OMe (5d)

image26

Sirupe Rdto: 78% (from 4d). Eluent: AcOEt: Hexane (1: 1). HPLC: t R = 10.73 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (400 MHz, CDCh): 5 7.36-7.14 (m, 10H, Ph, Z), 5.95 (d, 1H, J = 7.5 Hz, NH, Z), 5.10 (s, 2H, OCH2), 4.27 ( m, 1H, 3-H), 3.63 (s, 3H, OMe), 3.51 (t, 1H, J = 6.5 Hz, a-Phe), 2.95 (m, 2H, P-Phe), 2.73 (m, 1H , 1- H), 2.53 (m, 1H, 1-H), 1.97 (m, 1H, 2-H), 1.77 (m, 2H, 2-H), 1.44 (s, 9H, CH3 Bu). 13C NMR (75 MHz, CDCh): 174.5 (COO), 171.3 (COO), 156.0 (OCON), 136.9, 136.4,

129.0, 128.3, 127.9, 126.6 (Ar), 81.7 (C Bu), 66.6 (OCH2), 62.6 (Ca-Phe), 53.2 (C3), 51.5 (OMe), 44.1 (C1), 39.5 (Cp-Phe) , 31.7 (C2), 27.8 (CH3 Bu). MS (ES) +: 471.37 [M + H] +.

W - [(3S-Benzyloxycarbonylamino-3-ferc-butoxycarbonyl) prop-1-yl] -L-Ala-OMe (5e)

image27

GO2M6

dog

H

C02 * Bu

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Sirupe Rdto: 78% (from 4e). Eluent: AcOEt: Hexane (2: 1). HPLC: t R = 9.14 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (400 MHz, CDCfe): 5 7.33 (m, 5H, Z), 5.97 (d, 1H, J = 7.6 Hz, NH, Z), 5.12 (d, 1H, J = 12.0 Hz, OCH2), 5.06 (d, 1H, J = 12.0 Hz, OCH2), 4.30 (m, 1H, 3-H), 3.69 (s, 3H, OMe), 3.32 (q, 1H, J = 7.0 Hz, a-Ala), 2.73

(m, 1H, 1-H), 2.53 (m, 1H, 1-H), 1.97 (m, 1H, 2-H), 1.79 (m, 1H, 2-H, NH), 1.44 (s, 9H , CH3 fBu), 1.26 (d, 3H, J = 7.0 Hz, CH3 Ala). 13C NMR (75 MHz, CDCI3): 175.9 (COO),

171.5 (COO), 156.2 (OCON), 136.6, 128.6, 128.15, 128.1 (Ar), 82.0 (C fBu), 66.8 (OCH2), 56.4 (Ca-Ala), 53.4 (C3), 51.9 (OMe), 44.0 (C1), 32.1 (C2), 28.0 (CH3 fBu),

19.0 (CH3 Ala). MS (ES) +: 395.42 [M + H] +.

W- [4S - [(N-Benzyloxycarbonyl-N-methyl) amino-4-methoxycarbonyl] but-1-yl] -L-Phe-OlBu

(5f)

image28

C02 (Bu

Sirupe Rdto: 88% (from 4f). Eluent: AcOEt: Hexane (1: 1). HPLC: t R = 10.85 min (gradient from 5% to 100% of A, in 20 min). Proportion of rotamers M, m = 2: 1. 1H NMR (400 MHz, CDCh, majority rotamer): 5 7.37-7.19 (m, 10H, Ph, Z), 5.15 (s, 2H, OCH2), 4.78 (dd, 1H, J = 10.8, 5.1 Hz, 3- H), 3.70 (s, 3H, OMe), 3.41 (t, 1H, J = 7.0 Hz, a-Phe), 2.93 (m, 2H, p-Phe), 2.83 (s, 3H, NMe), 2.63 ( m, 2H, 1-H), 1.94 (m, 2H, 3- H), 1.73 (m, 2H, 2-H), 1.33 (s, 9H, CH3 fBu). 13C NMR (75 MHz, CDCl3): 173.8 (COO), 172.1 (COO), 157.1 (CON), 137.5, 129.5, 128.6, 128.3, 128.1, 127.9, 127.8, 126.6 (C, Ar), 81.3 (C fBu) , 67.6 (OCH2), 63.3 (Ca-Phe), 58.4 (C4), 52.2 (OMe), 47.4 (C1), 36.8 (Cp-Phe), 30.4 (NMe), 28.1 (CH3 * Bu), 26.6 (C2 ), 26.4 (C3). MS (ES) +: 499.44 [M + H] +. W - [(4S-ferc-Butoxycarbonylamino-4-benzyloxycarbonyl) but-1-yl] -H-L-Phe-OBn (5g)

image29

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Sirupe Rdto: 81% (from 4g). Eluent: AcOEt: Hexane (1: 2). HPLC: t R = 17.63 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (300 MHz, CDCI3): 5 7.74-6.87 (m, 15H, Ar), 5.62 (d, 1H, J = 8.4 Hz, NH), 5.22 (d, 1H, J = 12.4 Hz, OCH2), 5.16 (d, 1H, J = 12.4 Hz, OCH2), 5.09 (s, 2H, OCH2), 4.34 (m, 1H, 4-H), 3.55 (t, 1H, J = 7.5 Hz, a- Phe), 3.00 (m, 2H, p-Phe), 2.64 (m, 1H, 1-H), 2.44 (m, 1H, 1-H), 1.77 (m, 2H, 3-H), 1.46 (m, 9H, fBu ). 13C NMR (CDCI3): 5 174.5, 172.7 (COO), 155.6 (CON), 137.1,

135.6, 129.4, 128.9, 128.7, 128.6, 128.55, 128.5, 128.4, 128.35, 126.8 (Ar), 79.9 (C fBu), 67.0, 66.6 (OCH2), 62.9 (Ca-Phe), 53.5 (C4), 47.5 ( C1), 39.8 (Cp-Phe), 30.4 (C2),

28.5 (CH3 fBu), 25.8 (C3). MS (ES) +: 561 [M + H] +.

W - [(4S-Benzyloxycarbonylamino-4-methoxycarbonyl) but-1-yl] -L-Phe-O * Bu (5h)

image30

Sirupe Rdto: 89% (from 4h). HPLC: t R = 5.29 min (gradient of 15% to 95% of A, in 10 min). HPLC-MS: t R = 6.77 (gradient of 15% to 95% of A, in 10 min). 1H NMR (300 MHz, CDCI3): 5 7.26-7.08 (m, 10H, Ar), 6.03 (d, 1H, J = 7.8 Hz, 4-NH), 5.03 (s, 2H, OCH2), 4.05 (m, 1H, 4-H), 3.66 (s, 3H, OMe), 3.25 (dd, 1H, J = 7.7, 6.5 Hz, a-Phe), 2.87 (dd, 1H, J = 13.4, 6.4 Hz, P-Phe ), 2.75 (dd, 1H, J = 13.4, 7.7 Hz, P-Phe), 2.56 (m, 1H, 1- H), 2.35 (m, 1H, 1-H), 1.71 (m, 1H, 3- H), 1.48-1.38 (m, 3H, 3 and 2-H), 1.25 (s, 9H, CH3 fBu). MS (ES) +: 485.32 [M + H] +.

W - [(4S-Benzyloxycarbonylamino-4-methoxycarbonyl) but-1-yl] -L-Ala-O * Bu (5i)

HN ^ C02 (Bu

H

Y'

Or CO2M6

Sirupe Rdto: 77% (from 4i). HPLC-MS: t R = 5.20 min (gradient of 15% to 95% of A, in 10 min). 1H NMR (300 MHz, CDCI3): 5 7.26 (s, 5H, Ph), 6.15 (d, 1H, J = 7.7 Hz, 4-NH), 5.01 (s, 2H, OCH2), 4.28 (q, 1H, J = 7.7 Hz, 4-H), 3.65 (s, 3H, OMe), 3.06 (q, 1H, J = 6.9 Hz, a-Ala), 2.58 (m, 1H, 1'-H), 2.33 (dd , 1H, 1'-H), 1.76 (m, 2H, 3'-H), 1.45 (m, 2H, 2'-H), 1.37 (s, 9H, CH3 * Bu), 1.14 (d, 3H, J = 6.9 Hz, p-Ala). MS (ES) +: 409.33 [M + H] +.

W - [(2S-Benzyloxycarbomlammo-3-feml) prop-1-M] -L-Phe-OBn (5j)

image31

Sirupe Rdto: 99% (from 4j). Eluent: AcOEt: Hexane (1: 1). HPLC: t R = 15.41 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (400 MHz, CDCI3): 5 7.39-7.00 (m, 20H, Ar), 5.09 (s, 4H, OCH2), 5.08 (m, 1H, NH, Z), 3.87 (m, 1H, 2-H ), 3.49 (dd, 1H, 5 J = 7.9, 6.0 Hz, a-Phe), 2.99 (dd, 1H, J = 13.5, 6.0 Hz, 3-H), 2.86 (dd, 1H, J = 13.5, 7.9 Hz,

3-H), 2.79 (m, 1H, p-Phe), 2.70 (m, 2H, p-Phe, 1-H), 2.40 (dd, 1H, J = 12.3, 5.6 Hz, 1- H), 1.50 (sa, 1H, NH). 13C NMR (75 MHz, CDCI3): 5 173.7 (COO), 156.2 (CON), 137.7,

137.0, 136.7, 135.5, 129.4, 128.7, 128.6, 128.55, 128.5, 128.1, 127.0, 126.5 (Ar), 66.9,

66.7 (OCH2), 63.1 (Ca-Phe), 52.2 (C2), 49.8 (C1), 39.2 (C3), 37.5 (Cp-Phe). MS (ES) +: 10 523.28 [M + H] +.

W - [(2R-Benzyloxycarbomlammo-3-feml) prop-1-M] -L-Phe-OBn (5k)

image32

Sirupe Rdto: 93% (from 4k). Eluent: AcOEt: Hexane (1: 3). HPLC: t R = 15.53 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (400 MHz, CDCI3): 5 7.37-7.09 15 (m, 20H, Ar), 5.06 (s, 2H, OCH2), 5.04 (s, 2H, OCH2), 4.99 (sa, 1H, 2-NH) , 3.87 (m,

1H, 2-H), 3.45 (t, 1H, J = 6.9 Hz, a-Phe), 2.92 (dd, 1H, J = 13.5, 6.6 Hz, 3-H), 2.85 (m, 2H, 3-H , P-Phe), 2.64 (m, 2H, 1-H, P-Phe), 2.40 (dd, 1H, J = 12.3, 4.7 Hz, 1-H), 1.59 (sa, 1H, a-NH). 13C NMR (75 MHz, CDCI3): 5 174.5 (COO), 156.0 (CON), 137.8, 137.3, 136.8, 135.6, 129.5, 129.4, 128.7, 128.6, 128.55, 128.5, 128.1, 126.8, 126.5 (Ar), 66.7 , 20 66.65 (OCH2), 63.4 (Ca-Phe), 52.1 (C2), 49.8 (C1), 40.0 (C3), 39.0 (Cp-Phe). MS

(ES) +: 523.05 [M + H] +.

W - [(2R-Benzyloxycarbomlammo-3-feml) prop-1-M] -L-Ala-OBn (5l)

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Sirupe Rdto: 90% (from 4l). Eluent: AcOEt: Hexane (1: 3). HPLC: t R = 10.25 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (400 MHz, CDCh): 5 7.36-7.16 (m, 15H, Ar), 5.15 (d, 1H, J = 12.3 Hz, OCH2), 5.11 (m, 3H, NH, Z, OCH2), 5.10 ( d, 1H, J = 12.3 Hz, OCH2), 3.92 (m, 1H, 2-H), 3.30 (q, 1H, J = 7.0 Hz, a-Ala), 2.89 (dd, 1H, J = 13.9, 5.6 Hz, 3-H), 2.70 (dd, 1H, J = 13.9, 7.8 Hz, 3-H), 2.82 (dd, 1H, J = 12.0, 6.1 Hz, 1-H), 2.70 (dd, 1H, J = 12.0, 4.5 Hz, 1-H), 1.27 (d, 3H, J = 6.9 Hz, CH3 Ala). 13C NMR (75 MHz, CDCI3): 5 175.6 (COO), 156.1 (CON), 137.8, 136.7, 135.8, 129.5, 128.9, 128.7, 128.65, 128.6, 128.5, 128.3, 128.2, 126.6 (Ar), 66.7, 66.6 (2C, OCH2), 57.0 (Ca-Ala),

52.2 (C2), 49.9 (C1), 39.2 (C3), 19.4 (CH3 Ala). MS (ES) +: 447.36 [M + H] +. W - [(2S-DibencMammo-3-feml) prop-1 -il] -H-L-Phe-OBn (5m)

image34

Sirupe Rdto: 77% (from 4m). Eluent: AcOEt: Hex (1: 7). HPLC: t R = 7.23 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (400 MHz, CDCh): 5 7.28-6.87 (m, 25H, Ar), 4.96 (s, 2H, OCH2), 3.60 (d, 2H, J = 13.7 Hz, N-CH2), 3.43 (d, 2H, J = 13.7 Hz, N-CH2), 3.35 (dd, 1H, J = 7.8, 5.9 Hz, a-Phe), 2.87 (m, 2H, 3-H, 2-H), 2.85 (dd, 1H , J = 13.5, 5.8 Hz, P-Phe), 2.73 (dd, 1H, J = 13.5, 7.8 Hz, P-Phe), 2.50 (m, 2H, 1-H), 2.38 (dd, 1H, J = 14.9, 10.1 Hz Hz, 3-H). 13C NMR (75 MHz, CDCh): 5 174.0 (COO), 140.2, 139.9, 137.6, 135.8, 129.5, 129.4, 128.8, 128.6, 128.55, 128.5, 128.4, 127.0, 126.8,

126.0 (Ar), 66.5 (OCH2), 63.4 (Ca-Phe), 60.0 (C2), 53.9 (NCH2), 47.7 (C1), 39.5 (CP-Phe), 34.2 (C3). MS (ES) +: 569.27 [M + H] +. W - [(2S-DibencMammo-3-fenM) propM]] - L-Phe-OMe (5n)

image35

Sirupe Rdto: 85% (from 4n). Eluent: AcOEt: Hexane (1: 4). HPLC: t R = 4.15 min (gradient of 15% to 95% of A, in 5 min). 1H NMR (300 MHz, CDCh): 5 7.36-6.99 (m, 20H, Ar), 3.71 (d, 2H, J = 13.7 Hz, NCH2), 3.56 (s, 3H, OCH3), 3.50 (d, 2H, J = 13.7

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Hz, NCH2), 3.38 (dd, 1H, J = 8.2, 5.4 Hz, a-Phe), 3.30 (m, 2H, 2-H, 3-H), 2.93 (dd, 1H, J = 13.3, 5.4 Hz , 1-H), 2.78 (dd, 1H, J = 13.4, 8.2 Hz, 1-H), 2.55 (m, 2H, P-Phe), 2.42 (m, 1H, 3-H). 13C NMR (75 MHz, CDCI3): 5 174.8 (COO), 140.5, 140.2, 138.0, 129.8,

129.6, 129.1, 128.9, 128.7, 127.3, 127.2, 126.3 (C, Ar), 63.8 (a-Phe), 60.5 (C2), 54.2 (NCH2), 52.0 (OCH3), 48.0 (C1), 39.8 (P- Phe), 34.2 (C3). MS (ES) +: 494.10 [M + H] +. W - [(2S-Dibenzylamino-3-phenyl) propyl] -L-Phe-OfBu (5th)

image36

Sirupe Rdto: 88% (from 4th). Eluent: AcOEt: Hexane (1:10). HPLC: t R = 4.50 min (gradient of 15% to 95% of A, in 5 min). 1H NMR (300 MHz, CDCI3): 5 7.37-7.17 (m, 18H, Ar), 7.02 (m, 2H, Ar), 3.75 (d, 2H, J = 13.7 Hz, NCH2), 3.57 (d, 2H, J = 13.7 Hz, NCH2), 3.30 (dd, 1H, J = 7.9, 5.8 Hz, a-Phe), 3.00 (m, 2H, 2-H, 3-H), 2.90 (dd, 1H, J = 13.5 , 5.8 Hz, 1-H), 2.80 (dd, 1H, J = 13.5, 8.0 Hz, 1-H), 2.61 (m, 2H, P-Phe), 2.51 (m, 1H, 3-H), 1.33 (s, 9H, fBu). 13C NMR (75 MHz, CDCI3): 5 173.5 (COO), 140.4, 140.0,

138.0, 129.6, 129.4, 128.9, 128.5, 128.4, 126.9, 126.7, 125.9 (C, Ar), 81.0 (C, fBu),

64.0 (a-Phe), 60.3 (C2), 53.9 (NCH2), 47.5 (C1), 39.4 (P-Phe), 34.1 (C3), 28.1 (CH3, fBu). MS (ES) +: 536.16 [M + H] +.

Synthesis of N-alkyl-N-chloroacetyl amino acid derivatives

To a solution of the corresponding amino acid derivative (1.39 mmol, 0.765 g) in dry THF (5 mL) is added propylene oxide (20.85 mmol, 1.5 mL) and chloroacetyl chloride (1.67 mmol , 0.13 mL). After 1-2 h stirring at room temperature, the organic solvent evaporates to dryness. The resulting residue is purified on a silica gel column, using the eluent system indicated in each case. N-Chloroacetyl-N - [(3S-ferc-butoxycarbonylamino-3-benzyloxycarbonyl) prop-1-yl] -L- Phe-OMe (6a)

image37

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Sirupe Rdto: 92% (from 5a). Eluent: AcOEt: Hexane (1: 1). HPLC: t R = 15.70 min (gradient from 5% to 100% of A, in 20 min). Proportion of rotamers M, m = 9: 1. 1H NMR (400 MHz, CDCI3, majority rotamer): 5 7.35-7.09 (m, 10H, Ph), 5.18 (m, 1H, 3-NH), 5.16 (d, 1H, J = 12.0 Hz, OCH2), 5.07 (d, 1H, J = 12.0 Hz, OCH2), 4.16 (m, 1H, 3- H), 3.92 (m, 1H, a-Phe), 3.91 (d, 1H, J = 12.5 Hz, CH2CI), 3.86 (d, 1H, J = 12.5 Hz, CH2CI), 3.69 (s, 3H, OMe), 3.30 (m, 2H, p-Phe), 3.13 (m, 1H, 1-H), 2.70 (m, 1H, 1-H), 1.90 (m, 1H, 2-H), 1.52 (m, 1H, 2-H), 1.41 (s, 9H, CH3 fBu). 13C NMR (75 MHz, CDCI3, majority rotamer): 171.6 (COO), 170.4 (COO), 166.7 (CON), 155.4 (OCON), 137.8, 135.1, 129.4, 128.9, 128.75, 128.7, 127.0 (C, Ar) , 80.5 (C Bu), 67.6 (OCH2),

63.3 (Ca-Phe), 52.6 (OMe), 51.8 (C3), 47.0 (C1), 41.0 (CH2CI), 34.4 (Cp-Phe), 31.6 (C2), 28.4 (CH3 fBu). MS (ES) +: MS (ES) +: 569.30 [M + Na] +.

N-ChloroacetM-W - [(3S-ferc-butoxycarbomlammo-3-benzMoxicarboml) prop-1-M] -L- Phe-OBn (6b)

image38

Sirupe Rdto: 99% (from 5b). Eluent: AcOEt: Hexane (1: 2). HPLC: t R = 17.80 min (gradient from 5% to 100% of A, in 20 min). Rattan ratio M, m = 5: 2. 1H NMR (400 MHz, CDCI3, majority rotamer): 5 7.35-7.08 (m, 15H, Ar), 5.30 (d, 1H, J = 8.0 Hz, 3-NH), 5.15 (m, 2H, OCH2), 5.13 (d, 1H, J = 12.1 Hz, OCH2), 5.04 (d, 1H, J = 12.1 Hz, OCH2), 4.11 (m, 1H, a-Phe), 3.95 (m, 1H, 3-H), 3.88 (d, 1H, J = 12.4 Hz, CH2CI), 3.83 (d, 1H, J = 12.4 Hz, CH2CI), 3.31 (m, 2H, 1-H), 3.10 (dd, 1H, J = 15.6, 5.3 Hz , P-Phe), 2.73 (m, 1H, P-Phe), 1.86 (m, 2H, 2-H), 1.40 (s, 9H, CH3 Bu). 13C NMR (75 MHz, CDCI3): 171.5 (COO), 169.7 (COO), 166.6 (CON), 155.4 (OCON), 137.7,

135.6, 135.0, 129.4, 128.9, 128.7, 128.6, 128.4, 127.0 (C Ar), 80.5 (C Bu), 67.6 (OCH2), 67.4 (OCH2), 63.5 (Ca-Phe), 51.7 (C3), 47.0 ( CH2CI), 41.0 (C1), 34.4 (Cp - Phe), 31.6 (C2), 28.4 (CH3 fBu). MS (ES) +: 624.42 [M + H] +.

N-ChloroacetM-W - [(3S-ferc-butoxycarbomlammo-3-benzMoxicarboml) prop-1-M] -L- Ala-OMe (6c)

ci

image39

CO2M6

image40

‘C02Bn

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Sirupe Rdto: 80% (from 5c). Eluent: AcOEt: Hexane (1: 2). HPLC: t R = 13.15 min (gradient from 5% to 100% of A, in 20 min). Proportion of rotamers M, m = 5: 3. 1H NMR (400 MHz, CDCI3, majority rotamer): 5 7.36 (s, 5H, Ph), 5.47 (d, 1H, J = 7.6 Hz, 3-NH), 5.26 (d, 1H, J = 12.0 Hz, OCH2 ), 5.14 (d, 1H, J = 12.0 Hz, OCH2), 4.35 (m,

1H, 3-H), 4.16 (q, 1H, J = 7.0 Hz, a-Ala), 4.09 (s, 2H, CH2CI), 3.68 (s, 3H, OMe), 3.55 (m, 1H, 1-H ), 3.39 (t, 1H, J = 8.0 Hz, 1-H), 2.00 (m, 2H, 2-H), 1.44 (d, 3H, J = 7.0 Hz, CH3, Ala), 1.43 (s, 9H , CH3, fBu). 13C NMR (75 MHz, CDCI3): 171.55, 171.5 (COO),

166.5 (CON), 155.6 (OCON), 135.0, 129.0. 128.9, 128.8 (C Ar), 80.8 (C fBu), 67.8 (OCH2), 55.5 (Ca-Ala), 53.0 (C3), 52.5 (OMe), 44.7 (C1), 41.1 (CH2CI), 30.7 (C2) , 28.4 (CH3 fBu), 14.5 (CH3 Ala). MS (ES) +: 493.35 [M + Na] +.

N-Chloroacetyl-W - [(3S-benzyloxycarbonylamino-3-ferc-butoxycarbonyl) prop-1-yl] -L- Phe-OMe (6d)

image41

Sirupe Rdto: 90% (from 5d). Eluent: AcOEt: Hexane (1: 1). HPLC: t R = 15.66 min (gradient from 5% to 100% of A, in 20 min). Rotamer ratio M, m = 3: 1. 1H NMR (400 MHz, CDCh, majority rotamer): 5 7.37-7.13 (m, 10H, Ph), 5.46 (d, 1H, J = 7.2 Hz, 3-NH), 5.13 (d, 1H, J = 12.0 Hz , OCH2), 5.04 (d, 1H, J = 12.0 Hz, OCH2), 4.09 (m, 1H, 3-H), 3.99 (m, 1H, a-Phe), 3.99 (d, 1H, J = 12.0 Hz , CH2CO, 3.91 (d, 1H, J = 12.0 Hz, CH2Cl), 3.73 (s, 3H, OMe), 3.30 (m, 2H, -Phe), 3.20 (m, 1H, 1-H), 2.74 (m , 1H, 1- H), 1.88 (m, 1H, 2-H), 1.59 (m, 1H, 2-H), 1.42 (s, 9H, CH3 fBu) 13C NMR (75 MHz, CDCh): 171.3 , 170.3 (COO), 167.7 (CON), 156.0 (OCON), 137.6, 129.3, 128.7, 128.6,

128.4, 128.2, 126.9 (Ar), 83.0 (C fBu), 67.2 (OCH2), 63.2 (Ca-Phe), 52.9 (OMe), 52.6 (C3), 47.0 (CH2CO, 40.9 (C1), 34.3 (Cp- Phe), 31.5 (C2), 28.3 (CH3 fBu) MS (ES) +: 569.51 [M + Na] +.

N-Chloroacetyl-W - [(3S-benzyloxycarbonylamino-3-ferc-butoxycarbonyl) prop-1-yl] -L- Ala-OMe (6e)

Cl

Or I

CO2Me

image42

OR

O ^ "n

H

CO2tBu

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Sirupe Rdto: 92% (from 5e). Eluent: AcOEt: Hexane (1: 1). HPLC: t R = 13.54 min (gradient from 5% to 100% of A, in 20 min). Proportion of rotamers M, m = 2: 1. 1H NMR (400 MHz, CDCI3, majority rotamer): 5 7.35 (m, 5H, Ph), 5.58 (d, 1H, J = 7.0 Hz, 3-NH), 5.14 (d, 1H, J = 12.0 Hz, OCH2 ), 5.08 (d, 1H, J = 12.0 Hz, OCH2), 4.24 (m, 2H, 3-H, a-Ala), 4.12 (d, 1H, J = 12.5 Hz, CH2CI), 4.02 (d, 1H , J = 12.5 Hz, CH2CI), 3.70 (s, 3H, OMe), 3.42 (m, 2H, 1-H), 2.22 (m, 1H, 2-H), 1.99 (m, 1H, 2-H) , 1.48 (s, 9H, CH3 fBu), 1.47 (d, 3H, J = 6.4 Hz, CH3 Ala). 13C NMR (75 MHz, CDCI3): 171.4 (COO), 170.4 (COO), 166.6 (CON), 156.0 (OCON), 136.1, 133.2, 130.0, 128.6, 128.5, 128.3, 128.2,

128.0 (Ar), 83.1 (C fBu), 67.1 (OCH2), 55.4 (CD-Ala), 52.8 (C3), 52.4 (OMe), 44.6 (CH2CI), 41.0 (C1), 32.5 (C2), 27.9 ( CH3 fBu), 14.4 (CH3 Ala). MS (ES) +: 493.42 [M + Na] +.

W- [4S- (N-BencMoxicarbonM-N-metM) ammo-4-methoxycarboml] but-1-M] -N- chloroacetyl-L-Phe-O * Bu (6f)

image43

Sirupe Rdto: 94% (from 5f). Eluent: AcOEt: Hexane (1: 1). HPLC: t R = 16.21 min (gradient from 5% to 100% of A, in 20 min). Rattan ratio M, m = 5: 2. 1H NMR (400 MHz, CDCh, majority rotamer): 5 7.38-7.12 (m, 10H, Ph), 5.16 (s, 2H, OCH2), 4.68 (dd, 1H, J = 10.9, 5.2 Hz, a-Phe) , 4.39 (m, 1H, 4-H), 3.99 (d, 1H, J = 12.2 Hz, CH2CO, 3.94 (d, 1H, J = 12.5 Hz, CH2Cl), 3.69 (s, 3H, OMe), 3.28 ( m, 2H, 1-H), 2.89 (m, 1H, p-Phe), 2.77 (s, 3H, NCH3), 2.52 (m, 1H, P-Phe), 1.75-1.60 (m, 4H, 2- H, 3-H), 1.44 (s, 9H, CH3 fBu) .13C NMR (75 MHz, CDCh): 171.5 (COO), 168.9 (COO), 166.2 (CON), 157.1 (OCON), 138.4, 136.5, 129.4, 128.77, 128.7, 128.0, 126.8 (Ar), 82.0 (C fBu), 67.8 (OCH2), 64.2 (Ca-Phe), 57.6 (C4), 52.4 (OMe), 50.1 (C1), 41.1 (CH2CO,

34.4 (Cp-Phe), 30.4 (NCH3), 28.0 (CH3 fBu), 25.6 (C2), 24.9 (C3). MS (ES) +: 576.42 [M + H] +.

W - [(4S-ferc-butoxycarbonylamino-4-benzyloxycarbonyl) but-1-yl] -N-chloroacetyl-L-Phe-OBn (6g)

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Sirupe Rdto: 64% (from 5g). Eluent: AcOEt: Hexane (1: 3). Proportion of rotamers M / m = 6: 1. HPLC: t R = 17.68 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (300 MHz, CDCI3, majority rotamer): 5 7.38-7.12 (m, 15H, Ar), 5.18-5.04 (m, 4H, OCH2), 4.86 (d, 1H, J = 8.4 Hz, 4-NH) , 4.18 (m, 1H, 4-H), 3.94 (d, 1H, J = 12.5 Hz, CH2CI), 3.87 (d, 1H, J = 12.5 Hz, CH2CI), 3.82 (dd, 1H, J = 8.3, 6.9 Hz, a-Phe), 3.36 (m, 2H, p-Phe), 3.12 (dt, 1H, J = 15.4, 7.9 Hz, 1-H), 2.51 (dt, 1H, J = 15.3, 8.1 Hz, 1-H), 1.61 (m, 1H, 3H), 1.43 (m, 10H, fBu, 3-H), 1.24 (m, 2H, 2-H). 13C (CDCI3): 172.0, 169.7

(COO), 166.5 (CON), 155.4 (OCON), 137.9, 135.6, 135.5, 129.5, 129.6, 128.9, 128.8, 128.75, 128.7, 128.6, 128.5, 128.4, 127.0 (Ar), 80.3 (C fBu), 67.4 , 67.3 (OCH2), 63.5 (Ca-Phe), 52.7 (C4), 50.1 (CH2CI), 41.2 (C1), 34.4 (Cp-Phe), 29.7 (C3), 28.4 (CH3 fBu), 24.3 (C2) . MS (ES) +: 637.27 [M + H] +.

N - [(4S-BenzMoxicarbomlammo-4-methoxycarboml) but-1’-M] -N-doroacetM-L-Phe- O‘Bu (6h)

image45

N (S) C02'Bu

Sirupe Rdto: 94% (from 5h). HPLC: t R = 7.68 min (gradient from 30% to 95% of A, in 10 min). Rattan ratio M: m = 2: 1. 1H NMR (300 MHz, CDCl3): Major rotamer 5 7.36-7.13 (m, 10H, Ar), 5.27 (d, 1H, J = 8.3 Hz, 4-NH), 5.10 (s, 2H, OCH2),

4.27 (m, 1H, 4-H), 3.98 (m, 2H, CH2CI), 3.80 (m, 1H, a-Phe), 3.71 (s, 3H, OMe), 3.30 (m, 2H, P-Phe) , 3.17 (m, 2H, 1'-H), 3.61 (m, 1H, 1'-H), 1.85 (m, 1H, 3'-H), 1.68 (m, 3H, 3 ', - H 2' -H), 1.45 (s, 9H, CH3 fBu). MS (ES) +: 561.23 [M + H] +.

N - [(4S-Benzyloxycarbonylamino-4-methoxycarbonyl) but-1-yl] -N-Chloroacetyl-L-Ala- O‘Bu (6i)

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Sirupe Rdto: 70% (from 5i). HPLC: t R = 6.18 min (gradient from 30% to 95% of A, in 10 min). Proportion of rotamers M: m = 2: 1 1 H NMR (300 MHz, CDCl 3): majority rotamer 5 7.35 (s, 5H, Ph), 5.44 (sa, 1H, 4-NH), 5.11 (s, 2H, OCH2) , 4.40 (m, 1H, 4- H), 4.12 (s, 2H, CH2CI), 4.04 (m, 1H, a-Ala), 3.76 (s, 3H, OMe), 3.46 (m, 1H, 1'- H),

3.28 (m, 1H, 1'-H), 1.86 (m, 1H, 3'-H), 1.72 (m, 3H, 3'-H, 2'-H), 1.43 (s, 9H, CH3 fBu) ,

1.28 (d, 3H, J = 6.3 Hz, p-Ala). MS (ES) +: 485.16 [M + H] +.

N-ChloroacetM-W - [(2S-bencMoxicarbomlammo-3-feml) prop-1-M] -L-Phe-OBn (6j)

image47

Sirupe Rdto: 86% (from 5j). Eluent: AcOEt: Hexane (1: 2). HPLC: t R = 22.50 min (gradient from 30% to 95% of A, in 30 min). Rattan ratio M, m = 16: 1. 1H NMR (400 MHz, DMSO-d6, majority rotamer): 5 7.33-6.97 (m, 20H, Ar), 5.10 (d, 1H, J = 12.6 Hz, OCH2), 5.08 (sa, 1H, 2-NH) , 5.02 (d, 1H, J = 12.6 Hz, OCH2), 4.87 (d, 1H, J = 12.6 Hz, OCH2), 4.78 (d, 1H, J = 12.6 Hz, OCH2), 4.50 (d, 1H, J = 13.2 Hz, CH2CI), 4.33 (d, 1H, J = 13.2 Hz, CH2CI), 4.30 (dd, 1H, J = 9.4, 5.3 Hz, a-Phe), 3.72 (m, 1H, 2- H), 3.27 (dd, 1H, J = 13.8, 5.2 Hz, p-Phe), 3.19 (dd, 1H, J = 15.5, 6.4 Hz, 1-H), 3.10 (dd, 1H, J = 13.8, 9.4 Hz, P -Phe), 2.84 (dd, 1H, J = 13.5, 4.7 Hz, 3-H), 2.46 (m, 1H, 1-H), 2.40 (dd, 1H, J = 13.4, 6.8 Hz, 3-H) . 13C NMR (75 MHz, DMSO-de, majority rotamer): 5

169.4 (COO), 165.5 (CON), 155.7 (OCON), 138.2, 137.6, 136.9, 136.0, 129.3, 128.9,

128.5, 128.4, 128.3, 128.1, 127.8, 127.7, 127.6, 127.45, 126.4, 126.0 (Ar), 66.1, 65.2 (OCH2), 63.3 (Ca-Phe), 53.5 (C1), 52.8 (C2), 41.3 (CH2CI ), 37.4 (C3), 34.0 (Cp-Phe). MS (ES) +: 599.24 [M + H] +.

N-ChloroacetM-W - [(2R-bencMoxicarbomlammo-3-feml) prop-1-M] -L-Phe-OBn (6k)

image48

Sirupe Rdto: 83% (from 5k). Eluent: AcOEt: Hexane (1: 2). HPLC: t R = 22.71 min (gradient from 5% to 100% of A, in 20 min). Rotamer ratio M, m = 5: 1. 1H NMR (400 MHz, DMSO-d6, majority rotamer): 5 7.32-7.06 (m, 20H, Ar), 5.11 (sa, 1H, 2-NH), 5.11 (d, 1H, J = 12.6 Hz, OCH2) , 5.07 (d, 1H, J = 12.6 Hz, OCH2), 4.88 (s, 2H,

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OCH2), 4.37 (m, 1H, a-Phe), 4.28 (s, 2H, CH2CI), 3.85 (m, 1H, 2-H), 3.39 (m, 1H, □ - Phe), 3.30 (m, 1H , 1-H), 3.07 (dd, 1H, J = 13.8, 7.7 Hz, p-Phe), 2.80 (dd, 1H, J = 13.7,

4.5 Hz, 3-H), 2.75 (m, 1H, 1-H), 2.54 (m, 1H, 3-H). 13C NMR (75 MHz, DMSO-d6, majority rotamer): 5 169.5 (COO), 166.1 (CON), 155.6 (OCON), 138.4, 138.1, 137.1, 135.9, 129.4, 129.0, 128.3, 128.2, 128.0, 127.8, 127.6, 127.35, 126.3, 126.0 (Ar), 66.1, 64.9 (OCH2), 62.3 (Ca-Phe), 53.5 (C1), 51.7 (C2), 41.5 (CH2CI), 37.3 (C3),

34.5 (Cp-Phe). MS (ES) +: 599.15 [M + H] +.

N-ChloroacetM-W - [(2R-bencMoxicarbomlammo-3-feml) prop-1-M] -L-Ala-OBn (6l)

image49

Sirupe Rdto: 86% (from 5l). Eluent: AcOEt: Hexane (1: 3). HPLC: t R = 15.50 min (gradient from 5% to 100% of A, in 20 min). Proportion of rotamers M, m = 6: 1. 1H NMR (400 MHz, DMSO-d6, majority rotamer): 5 7.35-7.12 (m, 15H, Ar), 5.14 (sa, 1H, 2-NH), 5.09 (d, 1H, J = 12.2 Hz, OCH2) , 5.06 (d, 1H, J = 12.2 Hz, OCH2), 4.92 (d, 1H, J = 12.9 Hz, OCH2), 4.87 (d, 1H, J = 12.9 Hz, OCH2), 4.45 (d, 1H, J = 13.4 Hz, CH2CI),

4.28 (d, 1H, J = 13.4 Hz, CH2CI), 4.20 (q, 1H, J = 6.8 Hz, a-Ala), 3.95 (m, 1H, 2-H), 3.48 (dd, 1H, J = 15.1 , 9.2 Hz, 1-H), 3.40 (dd, 1H, J = 15.1, 5.0 Hz, 1-H), 2.82 (dd, 1H, J = 13.5, 4.2 Hz, 3-H), 2.58 (dd, 1H , J = 13.5, 10.3 Hz, 3-H), 1.38 (d, 3H, J = 6.8 Hz, CH3). 13C NMR (75 MHz, DMSO-da, majority rotamer): 5 171.0 (COO), 166.4 (CON), 156.3 (OCON), 138.6, 137.5, 136.3, 129.5, 128.75, 128.7, 128.5, 128.3, 128.0, 127.9, 127.4,

126.6 (Ar), 66.3, 65.3 (OCH2), 57.0 (Ca-Ala), 53.5 (C1), 51.8 (C2), 42.3 (CH2CI), 38.1 (C3), 14.5 (CH3). MS (ES) +: 523.28 [M + H] +.

N-Chloroacetyl-W - [(2S-dibenzylamino-3-phenyl-) prop-1-yl] -L-Phe-OBn (6m)

image50

Sirupe Rdto: 92% (from 5m). Eluent: AcOEt: Hex (1: 9). Rotamer ratio, M / m = 5: 1 HPLC: t R = 7.79 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (400 MHz, CDCh, majority rotamer): 5 7.57-6.52 (m, 25H, Ar), 5.10 (d, 1H, J = 12.2 Hz, OCH2), 4.95 (d, 1H, J = 12.2 Hz, OCH2 ), 4.17 (d, 1H, J = 12.2 Hz, CH2Cl), 3.86 (d, 1H, J = 12.2 Hz, CH-Cl), 3.56 (m, 4H, NCH2), 3.41 (dd, 1H, J = 15.8 , 4.8 Hz, 1'-

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H), 3.11 (m, 4-H, a-Phe, p-Phe, 2-H), 2.74 (dd, 1H, J = 13.2, 6.9Hz, 3'-H), 2.54 (dd, 1H, J = 13.3, 8.2 Hz, 3'-H), 2.31 (dd, 1H, J = 15.7, 5.8 Hz, 1'-H). 13C NMR (75 MHz, CDCh): 5

169.4 (COO), 167.1 (CON), 138.9, 137.8, 134.4, 133.6, 129.7, 129.3, 128.9, 128.8,

128.6, 128.4, 127.4, 126.6 (Ar), 67.3 (OCH2), 64.3 (a-Phe), 61.7 (C2 ’), 54.0 (NCH2),

52.6 (CH2CI), 50.0 (C1 ’), 41.6 (C3’), 34.8 (P-Phe). MS (ES) +: 646.23 [M + H] +. W-ChloroacetM-W - [(2S-dibenzylamm-3-feml) prop-1-M] -L-Phe-OMe (6n)

image51

Sirupe Rdto: 64% (from 5n). Eluent: AcOEt: Hexane (1: 6). HPLC: t R = 8.52 min (gradient from 30% to 95% of A, in 10 min). 1H NMR (400 MHz, CDCl3): 7: 1 5 rotamer mixture majority rotamer 7.29-6.89 (m, 20H, Ar), 4.33 (d, 1H, J = 11.7 Hz, CH2CI), 3.87 (d, 1H, J = 12.0 Hz, CH2CI), 3.64 (d, 2H, J = 13.7 Hz, NCH2), 3.51 (d, 2H, J = 13.7 Hz, NCH2), 3.47 (m, 1H, 1-H), 3.45 (s, 3H, OCH3), 3.14 (m, 1H, 2-H), 3.01 (m, 2H, P-Phe), 2.89 (dd, 1H, J = 13.2, 5.9 Hz, 3-H), 2.76 (dd, 1H , J = 9.8, 5.1 Hz, a-Phe), 2.46 (dd, 1H, J = 13.2, 9.0 Hz, 3-H), 2.25 (dd, 1H, J = 15.8, 4.9 Hz, 1-H). 13C NMR (75 MHz, CDCI3): 5 169.8 (COO), 166.8 (CON), 138.8, 138.6, 137.6, 129.7, 129.5, 128.7, 128.5, 128.4, 127.3, 126.6 (C, Ar), 63.6 (a-Phe ), 61.7 (C2), 54.0 (NCH2), 52.7 (C1),

52.0 (OMe), 41.4 (CH2CI), 34.1 (P-Phe), 33.9 (C3). MS (ES) +: 570.16 [M + H] +. W-Chloroacetyl-W - [(2S-dibenzylamino-3-phenyl) prop-1-yl] -L-Phe-OfBu (6o)

image52

Sirupe Rdto: 78% (from 5th). Eluent: AcOEt: Hexane (1: 6). HPLC: t R = 8.76 min (gradient from 30% to 95% of A, in 10 min). 1H NMR (400 MHz, CDCl3): mixture of 4: 1 5 rotamer majority 7.36-7.21 (m, 12H, Ar), 7.10-6.95 (m, 8H, Ar), 4.03 (d, 1H, J = 12.0 Hz , CH2CO, 3.78 (d, 1H, J = 12.0 Hz, CH2CO, 3.55 (d, 2H, J =

14.1 Hz, NCH2), 3.50 (d, 2H, J = 14.1 Hz, NCH2), 3.43 (dd, 1H, J = 15.4, 4.3 Hz, 1-H), 3.34 (m, 1H, a- Phe), 3.06 (m, 3H, 2-H, P-Phe), 2.76 (d, 2H, J = 7.5 Hz, 3-H), 2.54 (dd,

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1H, J = 15.5, 7.2 Hz, 1-H), 1.39 (s, 9H, fBu). 13C NMR (75 MHz, CDCI3): 5 168.4 (COO), 167.1 (CON), 139.2, 139.0, 138.3, 129.9, 129.5, 129.3, 128.8, 128.5, 128.2, 127.4, 126.7, 126.6 (C, Ar), 81.8 (C, fBu), 65.0 (Ca-Phe), 60.6 (C2), 53.9 (NCH2), 52.3 (C1), 41.4 (CH2CI), 35.2 (C3), 34.7 (Cp-Phe), 28.1 (CH3, fBu ). MS (ES) +: 612.14 [M + H] +.

Synthesis of N-alkyl-N-chloropropyl amino acid derivatives

To a solution of (R) - or (S) -2-doropropionic acid (1.74 mmol, 0.15 mL) in dry THF (5 mL), trichloroacetonitrile (2.32 mmol, 0.23 mL) is added . Then, it is cooled to 0 ° C with an ice bath and triphenylphosphine (1.96 mmol, 0.513 g) is slowly added and allowed to stir at room temperature for 1 h. After this time, the reaction is added to a solution of the corresponding N-alkyl derivative of amino acids (0.635 g, 1.16 mmol) and propylene oxide (17.4 mmol, 1.22 mL) in dry THF (2 mL) at 0 ° C. It is allowed to react at room temperature for 12h. After this time, the solvent is evaporated and the resulting residue is dissolved in AcOEt and washed successively with dric acid (10%), NaHCO3 (10%) and saturated NaCl solution. Finally, the organic residue is dried over anhydrous Na2SO4, filtered and evaporated to dryness. The reaction crude is purified on a silica gel column using the eluent system indicated in each case. W - [(3S-ferc-butoxycarbomlammo-3-benzyloxycarboml) prop-1-yl] -N- (2’S-chloropropanoyl) -L-Phe-OBn (7a)

image53

Sirupe Rdto: 68% (from 5b). Eluent: AcOEt: Hexane (1: 3). HPLC: t R = 10.64 min (gradient from 5% to 100% of A, in 20 min). Proportion of rotamers M, m = 5: 2. 1H NMR (400 MHz, CDCh, majority rotamer): 5 7.35-7.04 (m, 15H, Ar), 5.14 (d, 1H, J = 12.4 Hz, OCH2), 5.13 (s, 2H, OCH2), 5.03 (d , 1H, J = 12.1 Hz, OCH2), 4.93 (d, 1H, J = 6.5 Hz, 3-NH), 4.31 (q, 2H, J = 6.5 Hz, 1'-H), 4.06 (m, 1H, 3-H), 3.95 (m, 1H, a-Phe), 3.33 (dd, 1H, J = 14.1, 5.3, P-Phe), 3.26 (m, 1H, P-Phe), 3.00 (m, 2H, 1-H), 1.73 (m, 2H, 2-H), 1.59 (d, 3H, J = 6.4 Hz, 2'-H), 1.41 (s, 9H, CH3 fBu). 13C NMR (75 MHz, CDCl3): 171.8 (COO), 170.0 (COO), 169.4 (CON), 155.4 (OCON), 138.1, 135.8, 135.4,

129.6, 129.1, 129.0, 128.9, 128.7, 127.4, 127.2 (Ar), 80.8 (C fBu), 67.8, 67.7 (2C,

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OCH2), 63.8 (Ca-Phe), 51.7 (C3), 49.7 (CHCl), 46.6 (C1), 34.8 (Cp-Phe), 31.8 (C2),

28.6 (CH3 * Bu), 21.6 (C2 ’). MS (ES) +: 637.45 [M + H] +.

W - [(2S-BencMoxicarbomlammo-3-feml) prop-1-M] -N- (2’S-chloropropanoyl) -L-Phe- OBn (7b)

image54

Sirupe Rdto: 51% (from 5j). Eluent: AcOEt: Hexane (1: 4). HPLC: t R = 16.54 min (gradient from 5% to 100% of A, in 20 min). Proportion of rotamers M, m = 10: 1. 1H NMR (400 MHz, DMSO-d6, majority rotamer): 5 7.33-6.96 (m, 20H, Ar), 5.10 (d, 1H, J = 12.6 Hz, OCH2), 5.04 (d, 1H, J = 12.6 Hz , OCH2), 5.01 (q, 1H, J = 6.3 Hz, 1'-H), 4.87 (d, 1H, J = 12.6 Hz, OCH2), 4.81 (d, 1H, J = 12.6 Hz, OCH2), 4.39 (dd, 1H, J = 9.0, 5.8 Hz, a-Phe), 3.74 (m, 1H, 2-H), 3.33 (m, 1H, P-Phe), 3.13 (m, 2H, p-Phe, 1 -H), 2.90 (dd, 1H, J = 13.5, 4.5 Hz, 3-H), 2.64 (m, 1H, 1-H), 2.50 (m, 1H, 3-H), 1.51 (d, 3H, J = 6.4 Hz, 2'-H). 13C NMR (75 MHz, DMSO-de, majority rotamer): 5 169.4 (COO, CON),

155.7 (OCON), 138.3, 137.1, 136.8, 135.7, 129.2, 128.9, 128.3, 128.2, 128.1, 128.0, 128.0, 127.9, 127.7, 127.45, 126.4, 126.1 (Ar), 66.2, 65.2 (OCH2), 63.6 (Ca -Phe), 53.6 (C1), 52.9 (C2), 50.3 (CHCl), 36.8 (C3), 34.2 (Cp-Phe), 21.7 (C2 '). MS (ES) +: 613.29 [M + H] +.

W - [(2S-BencMoxicarbomlammo-3-feml) prop-1-M] -N- (2’R-chloropropanoyl) -L-Phe- OBn (7c)

image55

Sirupe Rdto: 50% (from 5j). Eluent: AcOEt: Hexane (1: 4). HPLC: t R = 17.39 min (gradient from 30% to 95% of A, in 20 min). Rotamer ratio M, m = 5: 1. 1H NMR (400 MHz, DMSO-d6, majority rotamer): 5 7.29-6.94 (m, 20H, Ar), 5.18 (d, 1H, J = 12.5 Hz, OCH2), 5.07 (q, 1H, J = 6.3 Hz , 1'-H), 4.91 (d, 1H, J = 12.5 Hz, OCH2), 4.84 (d, 1H, J = 12.6 Hz, OCH2), 4.74 (d, 1H, J = 12.6 Hz, OCH2), 4.21 (dd, 1H, J = 10.1,

4.7 Hz, a-Phe), 3.66 (m, 1H, 2-H), 3.23 (m, 2H, p-Phe, 1-H), 3.10 (dd, 1H, J = 13.6, 10.1 Hz, P-Phe ), 2.75 (dd, 1H, J = 13.5, 5.1 Hz, 3-H), 2.48 (m, 1H, 3-H), 2.22 (dd, 1H,

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J = 15.5, 5.9 Hz1-H), 1.42 (d, 3H, J = 6.3 Hz, 2’-H). 13C NMR (75 MHz, DMSO-d6, majority rotamer): 5 169.2 (COO), 168.7 (CON), 155.7 (OCON), 138.1, 137.4,

136.9, 136.1, 129.5, 128.9, 128.3, 128.2, 128.1, 127.9, 127.7, 127.4, 126.4, 126.1 (Ar),

65.9, 65.2 (OCH2), 63.2 (Ca-Phe), 53.2 (C1), 53.0 (C2), 40.0 (C1 ’), 37.6 (C3), 33.9 (Cp-Phe), 20.7 (C2’). MS (ES) +: 613.36 [M + H] +.

W - [(2R-BencMoxicarbomlammo-3-feml) prop-1-M] -N- (2’S-chloropropanoyl) -L-Phe- OBn (7d)

image56

Sirupe Rdto: 50% (from 5k). Eluent: AcOEt: Hexane (1: 4). HPLC: t R = 16.65 min (gradient from 5% to 100% of A, in 20 min). Proportion of rotamers M, m = 10: 1. 1H NMR (400 MHz, DMSO-d6, majority rotamer): 5 7.31-7.09 (m, 20H, Ar), 5.07 (s, 2H, OCH2), 4.97 (q, 1H, J = 6.4 Hz, 1'-H ), 4.90 (d, 1H, J = 12.6 Hz, OCH2), 4.86 (d, 1H, J = 12.6 Hz, OCH2), 4.40 (dd, 1H, J = 7.7, 5.9 Hz, a-Phe), 3.93 ( m, 1H, 2-H), 3.44 (m, 2H, p-Phe, 1-H), 3.13 (dd, 1H, J = 15.2, 8.4 Hz, 1-H), 3.00 (dd, 1H, J = 14.0, 5.9 Hz, p- Phe), 2.84 (dd, 1H, J = 13.6, 4.2 Hz, 3-H), 2.56 (m, 1H, 3-H), 1.43 (d, 3H, J = 6.3 Hz, 2'- H). 13C NMR (75 MHz, DMSO-de, majority rotamer): 5 169.7 (COO), 168.7 (CON),

155.7 (OCON), 138.6, 138.4, 137.1, 135.8, 129.4, 128.9, 128.3, 128.2, 128.1, 127.9,

127.8, 127.7, 127.35, 127.3, 126.2, 126.1 (Ar), 66.1.65.0 (OCH2), 62.2 (Ca-Phe), 53.5 (C1), 51.8 (C2), 49.7 (C1 '), 37.5 (C3), 35.0 (Cp-Phe), 21.3 (C2 '). MS (ES) +: 613.43 [M + H] +.

W - [(2R-BencMoxicarbomlammo-3-feml) prop-1-M] -N- (2’R-chloropropanoyl) -L-Phe- OBn (7e)

image57

Sirupe Rdto: 50% (from 5k). Eluent: AcOEt: Hexane (1: 3). HPLC: t R = 17.23 min (gradient from 30% to 95% of A, in 20 min). Rotation ratio M, m = 10: 1. 1H NMR (400 MHz, DMSO-da, major rotamer): 5 7.32-7.01 (m, 20H, Ar), 5.21 (d, 1H, J = 12.6 Hz, OCH2), 4.97 (d, 1H, J = 12.6 Hz , OCH2), 4.88 (s, 2H, OCH2), 4.54 (q, 1H,

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J = 6.4 Hz, 1'-H), 4.35 (dd, 1H, J = 9.5, 5.1 Hz, a-Phe), 3.75 (m, 1H, 2-H), 3.34 (m, 3H, p- Phe, 1-H), 3.14 (m, 1H, P-Phe), 2.66 (m, 1H, 3-H), 2.44 (dd, 1H, J = 15.9, 6.4 Hz, 3-H), 1.36 (d, 3H , J = 6.3 Hz, 2'-H) .13C NMR (75 MHz, DMSO-de, majority rotamer): 5 169.37 (COO), 168.34 (CON), 155.5 (OCON), 138.3, 137.6, 137.1, 135.9, 129.6, 129.0, 128.3, 128.2, 128.1, 128.0, 127.9, 127.7, 127.4, 126.5, 126.1 (Ar), 66.1, 65.1 (OCH2), 62.6 (Ca-Phe), 52.9 (C1), 52.2 (C2), 49.5 (C1 '), 37.1 (C3), 34.05 (Cp-Phe),

20.7 (C2 ’). MS (ES) +: 613.29 [M + H] +.

W - [(2S-dibencMammo-3-fenM-) prop-1-M] -N- (2’S-chloropropanoyl) -L-Phe-OBn (7f)

image58

Sirupe Rdto: 25% (from 5m). Eluent: AcOEt: Hex (1: 6). Proportion of rotamers M / m = 2: 1. HPLC: t R = 7.93 min (gradient from 5% to 100% of A, in 20 min). MS (ES) +: 659.19 [M + H] +. 1 H NMR (400 MHz, CDCI3, Major Rotary): 5 7,286.79 (m, 25H, Ar), 5.11 (d, 2H, J = 12.9 Hz, OCH2), 4.32 (q, 1H, J = 6.6 Hz, 1 '-H), 3.62 (dd, 1H, J = 9.5, 5.7 Hz, a-Phe), 3.55 (d, 2H, J = 13.9 Hz, NCH2), 3.47 (d, 2H, J = 13.9 Hz, NCH2) , 3.07 (m, 1H, 2-H), 3.31-3.16 (m, 4H, 3-H, 1-H, P-Phe), 2.90 (dd, 1H, J = 15.4,

8.4 Hz, 1-H), 2.65 (d, 2H, J = 13.2, 7.5 Hz, 3-H), 1.54 (d, 3H, J = 6.5 Hz, 2’-H). 13C NMR (75 MHz, CDCI3): 5 170.2 (COO), 169.6 (CON), 139.1, 138.9, 138.0, 135.7, 129.8, 129.4, 128.8, 128.75, 128.7, 128.5, 128.3, 127.3, 126.8, 126.5 (Ar) , 67.4 (OCH2), 64.0 (a-Phe), 59.1 (C2), 53.7 (NCH2), 52.0 (C1 '), 50.3 (C1), 34.8 (C3) 34.5 (P-Phe), 21.4 (C2') . MS (ES) +: 659.35 [M + H] +.

W - [(2S-DibencMammo-3-feml-) prop-1-M] -N- (2’R-chloropropanoyl) -L-Phe-OBn (7g)

image59

Sirupe Rdto: 52% (from 5m). Eluent: AcOEt: Hex (1: 6). Rotamer ratio M / m = 7: 1. HPLC: t R = 7.25 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (400 MHz, CDCI3, Major Rotary): 5 7.26-6.76 (m, 25H, Ar), 4.93 (s, 2H, OCH2), 4.58 (q, 1H, J = 6.5 Hz, 1'-H), 3.53 (dd, 1H, J = 15.9, 4.7 Hz, 1-H), 3.45 (m,

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4H, NCH2), 3.09 (dd, 1H, J = 13.8, 10.1 Hz, p-Phe), 3.03 (dd, 1H, J = 13.8, 4.6 Hz, p-Phe), 3.00 (m, 1H, 2-H ), 2.89 (dd, 1H, J = 10.1, 4.7 Hz, aH), 2.68 (dd, 1H, J = 13.2, 6.6 Hz, 3-H), 2.50 (dd, 1H, J = 13.1, 8.4, 3- H), 2.04 (dd, 1H, J = 15.9, 5.7 Hz, 1-H), 1.34 (d, 3H, J = 6.4 Hz, 2'-H). 13C NMR (75 MHz, CDCI3): 5 169.3 (COO), 169.2 (CON), 138.9,

137.8, 135.9, 129.9, 129.8, 128.75, 128.7, 128.5, 128.3, 127.4, 126.7, 126.6 (Ar), 67.1 (OCH2), 64.8 (a-Phe), 62.4 (C2), 53.9 (NCH2), 52.5 (C1 '), 49.6 (C1), 35.5 (C3), 34.4 (P-Phe), 20.7 (C2'). MS (ES) +: 659.21 [M + H] +.

W - [(2R-Benzyloxycarbonylamino-3-phenyl) prop-1-yl] - (2’S-chloropropanoyl) -L-Ala-OBn

(7h)

image60

Sirupe Rdto: 55% (from 5l). Eluent: AcOEt: Hexane (1: 4). HPLC: t R = 15.50 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (400 MHz, CDCI3): 5 7.35-7.10 (m, 15H, Ar), 5.10 (d, 1H, J = 12.8 Hz, OCH2), 5.03 (d, 1H, J = 12.8 Hz, OCH2), 5.06 (sa, 1H, 2-NH), 5.01 (q, 1H, J = 6.3 Hz, 1'-H), 4.90 (d, 1H, J = 12.9 Hz, OCH2), 4.85 (d, 1H, J = 12.9 Hz, OCH2), 4.20 (q, 1H, J = 6.7 Hz, a-Ala), 3.97 (m, 1H, 2-H), 3.64 (dd, 1H, J = 14.9, 10.5 Hz, 1-H), 3.40 (m, 1H, 1-H), 3.36 (s, 3H, OMe), 2.83 (dd, 1H, J = 13.6, 4.0 Hz, 3-H), 2.58 (dd, 1H, J = 13.6, 10.5 Hz , 3-H), 1.40 (d, 3H, J = 6.9 Hz, 2'-H), 1.37 (d, 3H, J = 6.3 Hz, CH3). 13C NMR (75 MHz, CDCI3): 5 170.6 (COO), 168.1 (CON), 155.8 (OCON), 138.3, 137.2, 136.0, 129.1, 128.2, 128.1, 127.8, 127.6, 127.2, 126.1 (Ar),

65.8, 64.8 (OCH2), 56.8 (Ca-Ala), 53.8 (C1), 51.2 (C2), 49.5 (C1 ’), 37.9 (C3), 20.8 (CH3) 14.5 (C2’). MS (ES) +: 537.05 [M + H] +.

W - [(2S-DibencMammo-3-feml) prop-1-M] -W- (2’S-doropropaneN) -L-Phe-OMe (7i)

image61

Sirupe Rdto: 49% (from 5n). Eluent: AcOEt: Hexane (1: 8). HPLC: t R = 7.92 min (gradient from 50% to 95% of A in 10 min). 1 H NMR (300 MHz, CDCl 3): mixture of rotamers 5: 1, 5 major rotamer 7.31-6.84 (m, 20H, Ar), 4.43 (q, 1H, J = 6.6 Hz, 1'-H), 3.64 (s , 3H, OCH3), 3.59 (m, 5H, a-Phe, NCH2), 3.34 (dd, 1H, J = 15.3, 3.9 Hz, 1-

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H), 3.23 (dd, 1H, J = 14.1, 5.8 Hz, p-Phe), 3.15 (dd, 1H,, J = 14.1, 9.4 Hz, p-Phe), 3.90 (m, 1H, 2-H) , 2.99 (dd, 1H,, J = 15.3, 8.1 Hz, 1-H), 2.77 (dd, 1H,, J = 13.4, 8.6 Hz, 3-H), 2.65 (dd, 1H, J = 13.4, 6.4 Hz, 3-H), 1.56 (d, 3H, J = 6.5 Hz, 2'-H). 13C NMR (75 MHz, CDCh): □ 170.3 (COO), 170.1 (CON), 139.07, 139.03, 137.9, 129.8, 129.3, 128.7,

128.6, 128.5, 128.4, 127.3, 126.7, 126.6 (C, Ar), 63.6 (a-Phe), 59.0 (C2), 53.6 (NCH2),

52.4 (OCH3), 51.8 (C1), 50.4 (C1 ’), 34.9 (H3), 34.8 (P-Phe), 21.4 (C2’). MS (ES) +: 584.39 (M + H +).

W - [(2S-dibencMammo-3-feml) prop-1-M] -W- (2’S-doropropaneN) -L-Phe-OtBu (7j)

image62

Sirupe Rdto: 80% (from 5th). Eluent: AcOEt: Hexane (1: 8). HPLC: t R = 5.61 min (gradient from 60% to 95% of A in 10 min). 1H NMR (400 MHz, DMSO-d6): 12: 1 rotamer mix, 5 major rotamer 7.30-6.93 (m, 20H, Ar), 4.09 (q, 1H, J = 6.5 Hz, 1'-H), 3.64 (dd, 1H, J = 9.5, 5.6 Hz, a-Phe), 3.56 (d, 2H, J = 13.8 Hz, NCH2), 3.45 (d, 2H, J = 13.8 Hz, NCH2), 3.25 (dd, 1H , J = 14.2 Hz, 1-H), 3.19 (dd, 1H, J = 14.2, 9.6 Hz, 1-H),

3.04 (m, 3H, p-Phe, 2-H), 2.82 (dd, 1H, J = 13.6, 4.9, 3-H), 2.65 (dd, 1H, J = 13.6, 9.3, 3- H), 1.54 (d, 3H, J = 6.5 Hz, 2'-H), 1.44 (s, 9H, CH3, fBu). 13C NMR (75 MHz, CDCI3): 5

169.9 (COO), 168.8 (CON), 139.3, 139.2, 139.1, 138.6, 129.8, 129.4, 128.8, 128.7, 128.3, 127.3, 126.7, 126.4 (C, Ar), 82.0 (C, Bu), 64.9 (a- Phe), 58.3 (C2), 53.7 (NCH2), 52.3 (C1), 49.9 (C1 '), 35.5 (P-Phe), 34.8 (C3), 28.1 (C2'), 21.3 (CH3, Bu). MS (ES) +: 626.35 (M + H +).

PREPARATION OF P-LACTAMIC DERIVATIVES

Intramolecular cyclization of chlorinated precursors, assisted by base

To a solution of the corresponding N-alkyl-N-chloroacetyl or N-chloropropionyl derivative (1.22 mmol, 0.754 g) in dry CH3CN (3 mL), under Ar atmosphere, BTPP (1.83 mmol, 0 is added , 56 mL) or Cs2CO3 (2.44 mmol, 0.793 g) and stirred at room temperature. When the reaction is complete the solvent is evaporated to dryness and the resulting residue is dissolved in AcOEt: H2O (1: 1) and the phases are separated. The organic extract is washed with saturated NaCl solution, dried over

Anhydrous Na2SO4 and evaporates to dryness. The reaction crude obtained is purified on a silica gel column, using the eluent system indicated in each case.

EXAMPLE 1

5 4R, S-Benzyl-4-methoxycarbonyl-1 - [(3’S-ferc-butoxycarbonylamino-3’-

benzyloxycarbonyl) prop-1’-il] -2-oxoazetidine

image63

Sirupe Rdto: 56% (from 6th, B: BTPP). Eluent: AcOEt: Hexane (1: 1). HPLC: t R = 17.22 min (gradient from 5% to 100% of A, in 20 min). HPLC (Chiral): tR = 19.02, 10 22.12 min (Isocratico 9/91 (Acetone / Hexane). Proportion of diastereoisomers M, m

= 80: 20. 1H NMR (400 MHz, CDCl3, majority diastereoisomer): 5 7.36-7.08 (m, 10H, Ph), 5.15 (sa, 3H, 3-NH, OCH2), 4.26 (m, 1H, 3'-H), 3.71 (s, 3H, OMe), 3.26 (d, 1H, J = 14.0 Hz, 4-CH2), 3.20 (m, 2H, 1'-H), 3.13 (d, 1H, J = 14.5 Hz. 4-CH2 ), 3.12 (d, 1H, J = 14.7 Hz, 3-H), 2.88 (d, 1H, J = 14.7 Hz, 3-H), 2.23 (m, 1H, 2'-H), 1.97 (m, 1H, 2'-H), 15 1.43 (s, 9H, CH3 Bu). 13C NMR (100 MHz, CDCh): 171.9 (COO), 166.4 (CON), 155.5

(OCON), 135.4, 134.5, 129.7, 128.9, 128.7, 128.6, 127.6 (C Ar), 80.1 (C Bu), 67.4 (OCH2), 63.0 (C4), 52.8 (OMe), 52.0 (C3 '), 45.7 (C3), 39.7 (C1 '), 38.7 (4-CH2), 31.0 (C2'), 28.4 (CH3 fBu) .MS (ES) +: 511.25 [M + H] +. Exact mass calculated for C28H34N2O7: 510.2366; Found: 510.2382.

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EXAMPLE 2

4R, S-Benzyl-4-methoxycarbonyl-1 - [(3’S-ferc-butoxycarbonylamino-3’-

Methyloxycarbonyl) prop-1’-yl] -2-oxoazetidine

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Sirupe Rdto: 5% (from 6th, B: BTPP). Eluent: AcOEt: Hexane (1: 1). HPLC: t R = 12.52, 12.65 min (gradient from 5% to 100% of A, in 20 min). HPLC (Chiral): tR = 17.17, 18.55 min (Isocratico 9/91 (Acetone / Hexane). Proportion of diastereo-isomers M, m = 86: 14. 1H NMR (400 MHz, CDCl3, majority diastereoisomer): 5 7.33- 7.12 (m, 5H, Ph), 5.19 (sa, 1H, 3-NH), 4.26 (m, 1H, 3'-H), 3.76 (s, 3H, OMe), 3.74 (s, 3H, OMe), 3.15 (d, 1H, J = 13.9 Hz, 4-CH2), 3.23 (m, 2H, 1'-H), 3.18 (d, 1H, J = 14.0 Hz, 4- CH2), 3.14 (d, 1H, J = 15.0 Hz, 3-H), 2.90 (d, 1H, J = 15.0 Hz, 3-H), 2.24 (m, 1H, 2'-H),

2.02 (m, 1H, 2’-H), 1.45 (s, 9H, CH3, * Bu). 13C NMR (100 MHz, CDCI3): 172.0 (COO), 166.5 (CON), 155.4 (OCON), 134.6, 129.8, 128.9, 128.8, 127.8 (C Ar), 80.3 (C fBu), 63.1 (C4), 52.9 (OMe), 52.7 (OMe), 51.9 (C3 '), 45.8 (C3), 40.0 (4-CH2), 38.9 (C1'), 31.0 (C2 '), 28.5 (CH3 fBu) .MS (ES) + : 435.32 [M + H] +. Exact mass calculated for C22H30N2O7: 434.20530; Found: 434.20662.

EXAMPLE 3

4R, S-Benzyl-4-benzyloxycarbonyl-1 - [(3’S-ferc-butoxycarbonylamino-3’-

benzyloxycarbonyl) prop-1’-il] -2-oxoazetidine

image65

Sirupe Rdto: 4% (from 6th, B: BTPP). Eluent: AcOEt: Hexane (1: 1). HPLC: t R = 17.22 min (gradient from 5% to 100% of A, in 20 min). HPLC (Chiralpak): tR = 20.15, 22.18 min (Isocratic 9/91 (Acetone / Hexane). Proportion of diastereoisomers M, m = 53:47. 1 H NMR (400 MHz, CDCh): 1 H NMR (400 MHz, CDCh) : 5 7.35-7.02 (m, 30H, Ar), 5.17 (d, 1H, J = 12.4 Hz, OCH2, M), 5.16 (s, 2H, OCH2, M), 5.14 (s, 4H, OCH2, m) , 5.11 (d, 1H, J = 12.4 Hz, OCH2, M), 5.03 (d, 1H, J = 6.6 Hz, 3-NH, m), 5.01 (m, 1H, 3-NH, M), 4.25 ( m, 1H, 3'-H, M), 4.21 (m, 1H, 3'-H, m), 3.25 (d, 2H, J = 14.2 Hz, 4-CH2, M, m), 3.24 (m, 2H, 1'-H, M), 3.15 (d, 1H, J = 14.8 Hz, 3-H, M), 3.14 (d, 1H, J = 14.2 Hz, 4-CH2, M), 3.13 (m, 3H, 1'-H, 4-CH2, m), 2.88 (d, 1H, J = 14.8 Hz, 3-H, M), 2.86 (d, 1H, J = 14.8 Hz, 3-H, m), 2.20 (m, 1H, 2'-H, m), 2.18 (m, 1H, 2'-H, M), 1.97 (m, 1H, 2'-H, M), 1.92 (m, 1H, 2 ' -H, m), 1.44 (s, 18H, CH3 fBu, M, m) 13C NMR (75 MHz, CDCh): 172.0 (COO, M), 171.9 (COO, m), 171.2 (COO, M, m ), 166.5 (COM, M), 166.4 (CON, m), 155.6 (OCON; M), 135.5, 135.4134.9, 134.8134.5, 134.4, 129.8, 129.0, 128.9, 128.8,

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128.7, 128.6, 128.5, 127.7, 127.6 (Ar), 80.3 (C fBu, M), 80.2 (C fBu, m), 67.9 (OCH2, M), 67.8 (OCH2, m), 67.5 (OCH2, M), , 67.4 (OCH2, m), 63.3 (C4, M), 63.1 (C4, m), 52.1 (C3 ', M), 51.9 (C3', m), 46.0 (C3, M), 45.8 (C3, m ), 40.1 (C1 ', M), 39.8 (C1', m), 39.0 (4-CH2, M), 38.9 (4-CH2, m), 31.1 (C2 ', M), 30.8 (C2', m ), 28.5 (CH3 fBu, M), 28.4 (CH3 fBu, m). 13C NMR (100 MHz, CDCI3): 172.0 (COO), 171.9 (COO), 171.2 (COO),

166.5 (CON), 166.4 (CON), 155.9 (OCON), 155.6 (OCON), 135.5, 135.4, 134.9, 134.8,

134.5, 134.4, 129.8, 129.0, 128.9, 128.8, 128.7, 128.6, 128.5, 127.7, 127.6 (Ar), 80.3 (C fBu), 80.2 (C fBu), 67.9, 67.8, 67.5, 67.4 (OCH2), 63.3 ( C4), 63.1 (C4), 52.1 (C3 '),

51.9 (C3 '), 46.0 (C3), 45.8 (C3), 40.1 (C1'), 39.8 (C1 '), 39.0 (4-CH2), 8.9 (4-CH2), 31.1 (C2'), 30.8 ( C2 '), 28.5 (CH3 fBu), 28.4 (CH3 fBu). MS (ES) +: 587.43 [M + H] +. Exact mass calculated for C34H38N2O7: 586.26790; Found: 586.26869.

EXAMPLE 4

4S-Benzyl-4-benzyloxycarbonyl-1 - [(3’S-ferc-butoxycarbonylamino-3’-

benzyloxycarbonyl) prop-1’-il] -2-oxoazetidine

image66

Sirupe Rdto: 29% (from 6b, B: BTPP). Semi-preparative HPLC purification (Chiral C). Eluent: Acetone: Hexane (13:87, tR = 9.78 min). HPLC: t R = 17.22 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (400 MHz, CDCh): 5 7.35-7.02 (m, 15H, Ar), 5.17 (d, 1H, J = 12.4 Hz, OCH2), 5.16 (s, 2H, OCH2), 5.11 (d, 1H, J = 12.4 Hz, OCH2), 5.01 (m, 1H, 3-NH), 4.25 (m, 1H, 3'-H), 3.25 (d, 1H, J = 14.2 Hz, 4-CH2), 3.24 (m , 2H, 1'-H), 3.15 (d, 1H, J = 14.8 Hz, 3-H), 3.14 (d, 1H, J = 14.2 Hz, 4-CH2), 2.88 (d, 1H, J = 14.8 Hz, 3-H), 2.18 (m, 1H, 2'-H), 1.97 (m, 1H, 2'-H), 1.44 (s, 9H, CH3 fBu). 13C NMR (75 MHz, CDCl3): 172.0 (COO), 171.2 (COO), 166.5 (CON), 155.6 (OCON), 135.5, 134.94, 134.5, 129.8, 129.0, 128.9, 128.8, 128.7, 128.5, 127.7 (C Ar), 80.3 (C fBu),

67.9, 67.5 (OCH2), 63.3 (C4), 52.1 (C3 ’), 46.0 (C3), 40.1 (C1’), 39.0 (4-CH2), 31.1 (C2 ’), 28.5 (CH3 fBu). MS (ES) +: 587.46 [M + H] +. Exact mass calculated for

C34H38N2O7: 586.26790; Found: 586.26869.

4R-Benzyl-4-benzyloxycarbonyl-1 - [(3’S-ferc-butoxycarbonylamino-3’-

benzyloxycarbonyl) prop-1’-il] -2-oxoazetidine

image67

Sirupe Rdto: 15% (from 6b, B: BTPP). HPLC semi-preparative purification 5 (Chiral C). Eluent: Acetone: Hexane (13:87, t R = 10.75 min). HPLC: t R = 17.22 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (400 MHz, CDCI3): 5 7.35-7.03 (m, 15H, Ar), 5.14 (s, 4H, OCH2), 5.03 (d, 1H, J = 6.6 Hz, 3-NH), 4.21 (m, 1H, 3'-H), 3.25 (d, 1H, J = 14.0 Hz, 4-CH2), 3.14 (d, 1H, J = 14.8 Hz, 3-H), 3.13 (m, 3H, 1'-H , 4- CH2), 2.86 (d, 1H, J = 14.8 Hz, 3-H), 2.20 (m, 1H, 2'-H), 1.92 (m, 1H, 2'-H), 1.44 (s, 9H, 10 CH3 fBu). 13C NMR (75 MHz, CDCI3): 171.9 (COO), 171.2 (COO), 166.4 (CON), 155.9

(OCON), 135.4, 134.8, 134.4, 129.8, 129.0, 128.9, 128.7, 128.6, 128. 5. 127.6 (Ar),

80.2 (C fBu), 67.8, 67.4 (OCH2), 63.1 (C4), 51.9 (C3 '), 45.8 (C3), 39.8 (C1'), 38.9 (4- CH2), 30.8 (C2 '), 28.4 ( CH3 fBu). MS (ES) +: 587.46 [M + H] +.

15 EXAMPLE 6

4R, S-Benzyl-4-benzyloxycarbonyl-1 - [(3’S-ferc-butoxycarbonylamino-3’-

benzyloxycarbonyl) prop-1’-il] - (3R, S>) - methyl-2-oxoazetidine

image68

Sirupe Rdto: 61% (from 7th, B: BTPP). Eluent: AcOEt: Hexane (1: 2). HPLC: 20 tR = 17.58 min (gradient from 5% to 100% of A, in 20 min). Proportion of

diastereoisomers M, m = 58: 42. 1H NMR (400 MHz, CDCI3): 5 7.37-7.04 (m, 15H, Ar), 5.22 (s, 2H, OCH2, M), 5.12 (s, 2H, OCH2, m), 5.03 (d, 1H, J = 6.6 Hz, 3-NH, M), 4.98 (d, 1H, J = 6.6 Hz, 3-NH, m), 4.14 (m, 1H, 3'-H), 3.51 (d, 1H, J = 15.1 Hz, 4-CH2), 3.45 (d, 1H, J = 14.5 Hz, 4-CH2), 3.03 (m, 4H, 4-CH2, 1'-H, 3-H), 2.24 (m, 1H, 2 '-H), 1.97 (m, 1H,

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2'-H), 1.43 (s, 9H, CH3 fBu), 1.08 (d, 3H, J = 7.6 Hz, 3-CH3, M), 1.07 (d, 3H, J = 7.6 Hz, 3- CH3, m ) .13C NMR (75 MHz, CDCI3): 172.0 (COO), 171.2 (COO), 171.1 (COO), 169.7 (CON), 169.6 (CON), 155.5 (OCON), 135.5, 135.0, 134.9, 134.8, 129.8 , 129.15, 129.1, 129.00, 128.95, 128.9, 128.8, 128.6, 128.5, 128.4, 128.3, 127.7, 127.6 (Ar), 79.9 (C fBu), 69.0 (C4), 67.7 (C4), 67.2, 67.1 (OCH2) , 54.0 (C3), 53.8 (C3), 52.1 (C3 '), 40.9 (4- CH2), 40.7 (4-CH2), 40.1 (C1'), 40.0 (C1 '), 30.8 (C2'), 30.7 (C2 '), 28.4 (CH3 fBu), 10.4 (3-CH3), 10.3 (3-CH3). MS (ES) +: 601.52 [M + H] +. Exact masses calculated for C35H40N2O7: 600.28355; Found: 600.28432.

EXAMPLE 7

4R, S-Methyl-4-methoxycarbonyl-1 - [(3’S-ferc-butoxycarbonylamino-3’-

benzyloxycarbonyl) prop-1’-il] -2-oxoazetidine

image69

Sirupe Rdto: 25% (from 6c, B: BTPP). Eluent: AcOEt: Hexane (1: 1). HPLC: t R = 12.64 min (gradient from 5% to 100% of A, in 20 min). Proportion of diastereoisomers M, m = 91: 9. 1H NMR (400 MHz, CDCl3, majority diastereoisomer): 5 7.35 (s, 5H, Ph), 5.24 (d, 1H, J = 7.8 Hz, 3-NH), 5.17 (s, 2H, OCH2), 4.30 (m , 1H, 3'-H), 3.74 (s, 3H, OMe), 3.26 (t, 2H, J = 7.9 Hz, 1'-H), 3.18 (d, 1H, J = 14.8 Hz, 3-H) , 2.79 (d, 1H, J = 14.8 Hz, 3-H), 2.20 (m, 1H, 2'-H), 2.00 (m, 1H, 2'-H), 1.43 (s, 12H, 4-CH3 fBu). 13C NMR (75 MHz, CDCI3): 172.8 (COO), 172.0 (COO), 166.3 (OCON), 135.4, 128.8. 128.6, 128.5 (Ar), 80.3 (C fBu), 67.4 (OCH2), 59.0 (C4), 58.9 (C3 '), 52.8 (OMe), 48.9 (C3), 37.9 (C1'), 30.9 (C2 ') , 28.4 (CH3 fBu), 20.5 (4-CH3). MS (ES) +: 435.32 [M + H] +.

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image70

Sirupe Rdto: 52% (from 6d, B: Cs2CO3). Eluent: AcOEt: Hexane (1: 1). HPLC: t R = 9.06, 9.31 min (gradient from 5% to 100% of A, in 20 min). HPLC (Chiral): tR = 22.22, 26.12 min (Isocratic 9/91 (Acetone / Hexane). Proportion of diastereoisomers M, m = 90: 10. 1 H NMR (400 MHz, CDCl3, majority diastereoisomer): 5 7.37-7.11 ( m, 10H, Ph), 5.44 (brs, 1H, 3-NH), 5.13 (d, 1H, J = 12.0 Hz, OCH2), 5.09 (d, 1H, J = 12.0 Hz, OCH2), 4.20 (m, 1H, 3'-H), 3.74 (s, 3H, OMe), 3.22 (m, 3H, 1'-H, 4-CH2),

3.16 (d, 1H, J = 12.0 Hz, 4-CH2), 3.14 (d, 1H, J = 14.8 Hz, 3-H), 2.89 (d, 1H, J = 14.8 Hz, 3-H), 2.23 ( m, 1H, 2'-H), 2.04 (m, 1H, 2'-H), 1.46 (s, 9H, CH3 fBu) .13C NMR (75 MHz, CDCI3): 171.9 (COO), 170.8 (COO) , 166.9 (CON), 156.0 (OCON), 136.4, 129.75,

129.7, 128.9, 128.6, 128.3, 128.2, 127.7 (Ar), 82.6 (C fBu), 67.0 (OCH2), 63.1 (C4),

52.9 (OMe, C3 ’), 45.8 (C3), 39.9 (4-CH2), 38.7 (C1’), 31.1 (C2 ’), 28.1 (CH3 * Bu). MS (ES) +: 511.55 [M + Na] +. Exact mass calculated for C28H34N2O7: 510.23660; Found: 510.23824.

EXAMPLE 9

4R, S-Methyl-4-methoxycarbonyl-1 - [(3’S-benzyloxycarbonylamino-3’-ferc-

butoxycarbonyl) prop-1’-il] -2-oxoazetidine

image71

Sirupe Rdto: 31% (from 6e, B: Cs2CO3). Eluent: AcOEt: Hexane (2: 1). HPLC: t R = 7.84 min (gradient from 5% to 100% of A, in 20 min). Proportion of diastereoisomers M, m = 69:31. 1H NMR (400 MHz, CDCl3, majority diastereoisomer): 5 7.35 (s, 5H, Ph), 5.48 (d, 1H, J = 7.9 Hz, 3-NH), 5.29 (s, 2H, OCH2), 4.24 (m , 1H, 3'-H), 3.76 (s, 3H, OMe), 3.50 (m, 1H, 1'-H), 3.26 (m, 1H, 1'-H), 3.18 (d, 1H, J = 14.9 Hz, 3-H), 2.78 (d, 1H, J = 14.9 Hz, 3-H), 2.16 (m, 1H, 2'-H), 1.99 (m, 1H, 2'- H), 1.46 ( s, 12H, CH3 fBu, 4-CH3). 13C NMR (75 MHz, CDCh): 172.7 (COO), 170.7

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(COO), 166.1 (CON), 156.0 (CON), 136.4, 128, 128.7, 128.25, 128.2 (C, Ar), 82.7 (C, * Bu), 67.1 (OCH2), 59.0 (C4), 52.9 (OMe , C3 '), 48.9 (C3), 37.9 (C1'), 31.3 (C2 '), 28.1 (CH3 * Bu), 20.8 (4-CH3). MS (ES) +: 457.44 [M + Na] +. Exact mass calculated for C30H30N2O7: 434.20530; Found: 434.20506.

EXAMPLE 10

4R, S-BenciM- [4’S- (W-BencMoxicarboml-W-metM) ammo-4’-methoxycarboml] but-1-

N] -4-ferc-butoxycarboml-2-oxoazetidma

image72

Sirupe Rdto: 77% (from 6f, B: BTPP). Eluent: AcOEt: Hexane (1: 1). HPLC: t R = 16.26 min (gradient from 5% to 100% of A, in 20 min). HPLC (Chiral): tR = 16.82, 17.83 min (Isocratico 10/90 (Acetone / Hexane). Rotamer ratio M, m = 60: 40. 1 H NMR (400 MHz, CDCh, majority rotamer): 5 7.36-7.13 ( m, 10H, Ph, Z), 5.15 (s, 2H, OCH2), 4.79 (dd, 1H, J = 10.1, 4.4 Hz, 4'-H), 3.71 (s, 3H, OMe), 3.29 (d, 1H, J = 15.1 Hz, 4-CH2), 3.25-3.10 (m, 4H, 1'-H, 3-H, 4-CH2), 3.06 (d, 1H, J = 14.0 Hz, 3-H), 2.85 (s, 3H, NMe), 1.94-1.71 (m, 4H, 2'-H, 3'-H), 1.41 (s, 9H, CH3 fBu). 13C NMR (75 MHz, CDCl3): 171.8 (COO ), 170.3 (COO), 166.9 (CON), 157.1 (OCON), 136.7, 135.1, 129.8,

128.7, 128.6, 128.1, 128.0, 127.9, 127.5 (Ar), 83.0 (C fBu), 67.6 (OCH2), 63.4 (C4),

58.4 (C4 '), 52.3 (OMe), 45.8 (C3), 41.4 (C1'), 40.0 (4-CH2), 30.7 (NMe), 27.9 (CH3 fBu), 26.6 (C3 '), 25.5 (C2' ). MS (ES) +: 539.67 [M + H] +. Exact mass calculated for C30H38N2O7: 538.26790; Found: 538.26910.

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C02 * Bu

image73

A solution of the corresponding compound of Example 10 (0.15 mmol, 82 mg) in MeOH (15 mL) is hydrogenated at room temperature and 15 psi of pressure for 4 h, using Pd-C (16 mg, 20%) as catalyst. . Once the catalyst was separated by filtration, the solvent was evaporated to dryness, the reaction crude was purified by column chromatography using as eluents MeOH: DCM in proportion (1:10), obtaining a 48 mg (78%) syrup of Product described below. HPLC: t R = 8.87 min (gradient from 5% to 100% of A, in 20 min). Proportion of diastereoisomers M, m = 73: 27. 1H NMR (400 MHz, CDCl3, majority diastereoisomer): 5 7.31-7.17 (m, 5H, Ph), 3.74 (s, 3H, OMe), 3.32 (d, 1H, J = 13.9 Hz, 4- CH2), 3.21 (m, 3H, 1'-H, 4'-H), 3.16 (d, 1H, J = 14.8 Hz, 3-H), 3.10 (d, 1H, J = 13.9 Hz, 4- CH2), 2.86 ( d, 1H, J = 14.8 Hz, 3-H), 2.39 (s, 3H, NMe), 1.69 (m, 4H, 3'-H, 2'-H), 1.42 (s, 9H, CH3 fBu). 13C NMR (75 MHz, CDCl3, majority diastereoisomer): 174.8 (COO), 169.8 (COO), 166.4 (CON), 134.6, 129.3, 128.2, 127.0 (Ar), 82.5 (C fBu), 62.9 (C4),

62.3 (C4 ’), 51.4 (OMe), 45.3 (C3), 41.2 (C1’), 39.7 (4-CH2), 34.3 (NMe), 30.3 (C3 ’),

27.5 (CH3 * Bu), 24.7 (C2 ’). MS (ES) +: 405.49 [M + Na] +. Exact mass calculated for C22H32N2O5: 404.23112; Found: 404.23245.

EXAMPLE 12

4R, S-Benzyl-4-benzyloxycarbonyl-1 - [(4’S-ferc-butoxycarbonylamino-4’-

benzyloxycarbonyl) but-1’-yl] -2-oxoazetidine

image74

Sirupe Rdto: 71% (from 6g). Proportion of diastereoisomers, M / m = 5: 1. Eluent: AcOEt: Hexane (1: 4). HPLC: t R = 17.18 min (gradient from 5% to 100% of A, in 20 min). 1 H NMR (400 MHz, CDCl 3, majority diastereoisomer): 5 7.40-6.90 (m,

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15H, Ar), 5.17-4.95 (m, 5H, OCH2, 4-NH), 4.22 (m, 1H, 4'-H), 3.24 (d, 1H, J = 14.0 Hz, 4-CH2), 3.23 ( d, 1H, J = 14.0 Hz, 4-CH2), 3.11 (d, 1H, J = 14.8 Hz, 3-H), 3.04 (m, 2H, 1'-H), 2.82 (d, 1H, J = 14.8 Hz, 3-H), 1.60 (m, 2H, 3'-H), 1.52 (m, 2H, 2'-H), 1.36 (s, 9H, fBu). 13C NMR (CDCl3): 5 1725, 171.1 (COO), 166.4 (C2), 155.5 (OCON), 135.5, 134.9, 134.6, 129.7, 128.9, 128.8, 128.7, 128.7,128.5, 128.4, 127.6 (Ar), 80.0 (C fBu),

67.7, 67.2 (OCH2), 63.0 (C4), 53.4 (C4 ’), 45.8 (C3), 41.7 (C3), 40.1 (4-CH2), 30.3 (C3’),

28.5 (CH3 * Bu), 24.6 (C2 ’). MS (ES) +: 623 [M + Na] +.

EXAMPLE 13

4R, S-Benzyl-N - [(4’S-benzyloxycarbonylamino-4’-methoxycarbonyl) but-1’-il] -4-ferc-

butoxycarbonyl-2-oxoazetidine

image75

Sirupe Rdto: 78% (from 6h). HPLC-MS: t R = 5.51 min (gradient from 50% to 95% of A, in 15 min). HPLC (chiral): tR = 19.85, 22.39 min (isocratic: 11/89: acetone / hexane), proportion of diastereoisomers (4R, 4’S) :( 4S, 4’S), M: m = 5: 1. 1H NMR (300 MHz, CDCI3): majority isomer 5 7.35-7.07 (m, 10H, Ar), 5.46 (d, 1H, J = 7.8, 4-NH), 5.04 (s, 2H, OCH2), 4.30 (m , 1H, 4'-H), 3.67 (m, 3H, OCH3), 3.22 (d, 1H, J = 14.0, 4-CH2), 3.13-3.01 (m, 3H, 1'-H, 4-CH2, 3-H), 3.80 (d, 1H, J = 14.7, 3-H), 1,801.56 (m, 4H, 2'-H, 3'-H), 1.35 (s, 9H, CH3 fBu). 13C NMR (100 MHz, CDCI3): majority isomer 172.7 (COO), 170.4 (COO), 166.9 (C2), 156.1 (OCON), 136.4, 135.0, 129.8, 128.8, 128.6, 128.3, 128.2, 127.5 (Ar), 83.1 (C fBu), 67.1 (OCH2), 63.5 (C4), 53.7 (C4 '), 52.6 (OMe), 45.9 (C3), 41.5 (C1'), 40.0 (4-CH2), 29.8 (C3 ') , 28.0 (fBu), 24.6 (C2 '). MS (ES) +: 525.24 [M + H] +.

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image76

Sirupe Rdto: 58% (from 6i). HPLC-MS: t R = 3.06 min (gradient from 50% to 95% of A, in 15 min). Proportion of diastereoisomers (4R, 4’S) :( 4S, 4’R), M: m = 2: 1. 1H NMR (300 MHz, CDCI3): majority isomer 5 7.37-7.31 (m, 5H, Ph), 5.54 (d, 1H, J = 7.1 Hz, 4-NH), 5.11 (s, 2H, OCH2), 4.37 ( m, 1H, 4'-H), 3.75 (s, 3H, OMe), 3.20 (m, 2H, 1'- H), 3.14 (d, 1H, J = 14.5 Hz, 3-H), 2.76 (d , 1H, J = 14.5 Hz, 3-H), 1.90 (m, 1H, 3'-H), 1.80-1.63 (m, 3H, 3'-H, 2'-H), 1.59 (s, 3H, 4-CH3), 1.45 (s, 9H, CH3 fBu). MS (ES) +:

449.16 [M + H] +, 471.46 [M + Na] +.

EXAMPLE 15

4R, S-BencM-4-bencMoxicarbomM - [(2’S-bencMoxicarbomlammo-3’-feml) prop-1’-

il] -2-oxoazetidine

image77

Sirupe Rdto: 11% (from 6j, B: Cs2CO3). Eluent: AcOEt: Hexane (1: 3). HPLC: t R = 20.74 min (30% gradient to 95% A, in 30 min). Proportion of diastereoisomers (4S, 2’S) / (4R, 2’S) M / m = 83: 17. 1 H NMR (500 MHz, CDCI3,

Major diastereoisomer): 5 7.34-6.98 (m, 20H, Ar), 5.48 (d, 1H, J = 8.5 Hz, 2-NH), 5.15 (d, 1H, J = 12.0 Hz, OCH2), 5.09 (d, 1H, J = 12.0 Hz, OCH2), 5.06 (d, 1H, J = 12.5 Hz, OCH2), 5.02 (d, 1H, J = 12.5 Hz, OCH2), 4.15 (m, 1H, 2'-H), 3.37 (dd, 1H, J = 14.5, 8.1 Hz, 1'-H), 3.27 (d, 1H, J = 14.0 Hz, 4-CH2), 3.22 (d, 1H, J = 14.9 Hz, 3-H) , 3.18 (dd, 1H, J = 14.5, 4.3 Hz, 1'-H), 2.98 (d, 1H, J = 14.0 Hz, 4-CH2), 2.90 (d, 1H, J = 14.9 Hz, 3-H ), 2.85 (dd, 1H, J = 14.1, 7.6 Hz, 3'-H), 2.76 (dd, 1H, J = 14.1, 6.8 Hz, 3'-H). 13C NMR (125 MHz, CDCl3, majority diastereoisomer): 5 170.8 (COO), 167.4 (C2), 156.1 (OCON), 137.4, 134.9, 134.2, 129.7, 129.4, 128.9, 128.8, 128.7, 128.6, 128.1, 128.0, 127.6, 126.8 (Ar), 67.9, 67.8 (OCH2), 63.2 (C4), 51.6 (C2 '), 45.4 (C3), 45.2 (C1'), 39.7

(4-CH2), 39.1 (C3 ’). MS (ES) +: 563.46 [M + H] +. Exact mass calculated for C35H34N2O5: 562.24677; Found: 562.24804.

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EXAMPLE 16

4R, S-BencM-4-bencMoxicarbomM - [(2’R-bencMoxicarbomlammo-3’-feml) prop-1’-

il] -2-oxoazetidine

image78

Sirupe Rdto: 11% (from 6k, B: Cs2CO3). Eluent: AcOEt: Hexane (1: 3). HPLC: t R = 20.70 min (30% gradient to 95% of A, in 30 min). Proportion of diastereoisomers (4S, 2’S) / (4R, 2’S) M / m = 91: 9. 1 H NMR (400 MHz, CDCI3,

Major diastereoisomer): 5 7.34-6.99 (m, 20H, Ar), 5.54 (d, 1H, J = 8.5 Hz, 2-NH), 5.14 (d, 1H, J = 12.0 Hz, OCH2), 5.06 (s, 2H, OCH2), 5.04 (d, 1H, J = 12.0 Hz, OCH2),

4.16 (m, 1H, 2'-H), 3.29 (d, 1H, J = 14.0 Hz, 4-CH2), 3.22 (m, 3H, 3-H, 1'-H), 3.15 (d, 1H, J = 14.0 Hz, 4-CH2), 2.89 (d, 1H, J = 14.9 Hz, 3-H), 2.78 (dd, 1H, J = 14.0, 5.5 Hz, 3'- H), 2.72 (dd, 1H , J = 14.0, 6.8 Hz, 3'-H). 13C NMR (75 MHz, CDCI3, majority diastereoisomer): 5 170.9 (COO), 167.1 (C2), 156.2 (OCON), 137.2, 134.8, 134.5, 129.6, 129.4, 128.9, 128.8, 128.6, 128.5, 128.1, 127.5, 126.7 (Ar), 67.8, 66.6 (OCH2), 63.3 (C4), 51.5 (C2 '), 45.8 (C3), 45.7 (C1'), 40.3 (4-CH2), 39.1 (C3 '). MS (ES) +: 563.46 [M + H] +. Exact mass calculated for C35H34N2O5: 562.24677; Found: 562.24834.

EXAMPLE 17

4S-Benzyl-4-benzyloxycarbonyl-3S-methyl-1 - [(2’S-benzyloxycarbonylamino-3’-

phenyl) prop-1’-il] -2-oxoazetidine

image79

Sirupe Rdto: 65% (from 7b, B: BTPP). Eluent: AcOEt: Hexane (1: 2). HPLC: t R = 16.20 min (gradient from 30% to 95% of A, in 20 min). Proportion of isomers M (3S, 4S, 2’S): m (3R, 4R, 2’S) = 97: 3. 1H NMR (400 MHz, CDCI3, majority isomer): 5 5 7.35-6.96 (m, 20H, Ar), 5.75 (d, 1H, J = 7.4 Hz, 2-NH), 5.24 (d, 1H, J = 12.0 Hz, OCH2),

5.14 (d, 1H, J = 12.0 Hz, OCH2), 5.05 (s, 2H, OCH2), 4.17 (m, 1H, 2'-H), 3.43 (d, 1H, J = 14.5 Hz, 4-CH2) , 3.23 (dd, 1H, J = 14.5, 8.0, 1'-H), 3.11 (q, 1H, J = 7.6 Hz, 3-H), 3.05 (d, 1H, J = 14.5 Hz, 4-CH2) , 3.00 (dd, 1H, J = 14.5, 4.0 Hz, 1'-H), 2.86 (dd, 1H, J = 14.0, 8.0 Hz, 3'-H), 2.72 (dd, 1H, J = 14.0, 7.0 Hz, 3'-H), 1.08 (d, 3H, J = 7.5 Hz, 3-CH3) .13C 10 NMR (75 MHz, CDCI3, majority isomer): 5 171.01 (COO), 170.57 (C2), 156.03 ( OCON), 137.8, 137.0, 134.9, 134.8, 129.7, 129.2, 128.9, 128.85, 128.8, 128.7, 128.5, 128.45, 127.9, 127.8, 127.5, 126.5 (Ar), 68.68 (C4), 67.67, 66.27 (OCH2), 53.84 (C3), 51.89 (C2 '), 46.06 (C1'), 40.69 (4-CH2). 39.10 (C3 ’), 10.60 (3-CH3). MS (ES) +: 577.3 [M + H] +.

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EXAMPLE 18

4R-BencM-4-bencMoxicarbonM-3R-metiM - [(2’S-bencMoxicarbomlammo-3’-

phenyl] prop-1’-il] -2-oxoazetidine

image80

20 Sirupe. Rdto: 10% (from 7c, B: BTPP). Eluent: AcOEt: Hexane (1: 3). HPLC: t R = 16.36 min (gradient from 30% to 95% of A, in 20 min). Proportion of isomers M (3R, 4R, 2’S): m (3S, 4S, 2’S) = 85:15. 1H NMR (400 MHz, CDCI3, majority isomer): 5 7.37-6.99 (m, 20H, Ar), 5.83 (d, 1H, J = 8.1 Hz, 2-NH), 5.27 (d, 1H, J = 12.0 Hz , OCH2),

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5.17 (d, 1H, J = 12.0 Hz, OCH2), 5.05 (s, 2H, OCH2), 4.07 (m, 1H, 2'-H), 3.56 (d, 1H, J = 14.5 Hz, 4-CH2) , 3.43 (q, 1H, J = 7.6 Hz, 3-H), 3.03 (d, 1H, J = 14.5 Hz, 4-CH2), 2.99 (m, 2H, 1'-H), 2.71 (dd, 1H , J = 13.5, 7.4 Hz, 3'-H), 2.64 (dd, 1H, J = 13.4, 6.6 Hz, 3'-H), 1.08 (d, 3H, J = 7.5 Hz, 3-CH3). 13C NMR (100 MHz, CDCI3, majority isomer): 5

171.3 (COO), 170.3 (C2), 156.3 (OCON), 137.5, 137.1, 135.1, 134.8, 129.8, 129.5, 129.45, 129.4, 129.2, 129.1, 128.95, 128.9, 128.85, 128.8, 128.6, 128.55, 128.5, 128.0 , 127.6, 126.6 (Ar), 69.1 (C4), 67.9, 66.4 (OCH2), 54.2 (C3), 51.6 (C2 '), 47.1 (C1'), 41.3 (4-CH2). 39.4 (C3 ’), 10.6 (3-CH3). MS (ES) +: 577.3 [M + H] +.

EXAMPLE 19

4R-BencM-4-bencMoxicarbonM-3R-metiM - [(2’R-bencMoxicarbomlammo-3’-

phenyl) prop-1’-il] -2-oxoazetidine

image81

Sirupe Rdto: 44% (from 7e, B: BTPP). Eluent: AcOEt: Hexane (1: 3). HPLC: t R = 16.20 min (gradient from 30% to 95% of A, in 20 min). Proportion of isomers M (3R, 4R, 2’R): m (3S, 4S, 2’R) = 77: 23. 1H NMR (400 MHz, CDCI3, majority diastereoisomer): 5 7.35-6.96 (m, 20H, Ar), 5.75 (d, 1H, J = 7.4 Hz, 2-NH), 5.24 (d, 1H, J = 12.0 Hz , OCH2), 5.14 (d, 1H, J = 12.0 Hz, OCH2), 5.05 (s, 2H, OCH2), 4.17 (m, 1H, 2'- H), 3.43 (d, 1H, J = 14.5 Hz, 4-CH2), 3.23 (dd, 1H, J = 14.5, 8.0, 1'-H), 3.11 (q, 1H, J = 7.6 Hz, 3-H), 3.05 (d, 1H, J = 14.5 Hz, 4-CH2), 3.00 (dd, 1H, J = 14.5, 4.0 Hz, 1'-H), 2.86 (dd, 1H, J = 14.0, 8.0 Hz, 3'-H), 2.72 (dd, 1H, J = 14.0, 7.0 Hz, 3'-H), 1.08 (d, 3H, J = 7.5 Hz, 3- CH3) .13C NMR (75 MHz, CDCl3, majority diastereoisomer): 5 171.01 (COO), 170.57 (C2) , 156.03 (OCON), 137.8, 137.0, 134.9, 134.8, 129.7, 129.2, 128.9, 128.85, 128.8, 128.7, 128.5, 128.45, 127.9, 127.8, 127.5, 126.5 (Ar), 68.68 (C4), 67.67, 66.27 ( OCH2), 53.84 (C3), 51.89 (C2 '), 46.06 (C1'), 40.69 (4-CH2). 39.10 (C3 ’), 10.60 (3- CH3). MS (ES) +: 577.18 [M + H] +. Exact mass calculated for C36H36N2O5: 576.26242; Found: 576.26364.

EXAMPLE 20

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4S-4-BencM-4-bencMoxicarboml-3S-metiM - [(2’R-bencMoxicarbomlammo-3’-

phenyl) prop-1’-il] -2-oxoazetidine

image82

Sirupe Rdto: 39% (from 7d, B: BTPP). Eluent: AcOEt: Hexane (1: 3). HPLC: t R = 16.36 min (gradient from 30% to 95% of A, in 20 min). 1H NMR (400 MHz, CDCl3): 5 7.37-6.99 (m, 20H, Ar), 5.83 (d, 1H, J = 8.1 Hz, 2-NH), 5.27 (d, 1H, J = 12.0 Hz, OCH2) ,

5.17 (d, 1H, J = 12.0 Hz, OCH2), 5.05 (s, 2H, OCH2), 4.07 (m, 1H, 2'-H), 3.56 (d, 1H, J = 14.5 Hz, 4-CH2) , 3.43 (q, 1H, J = 7.6 Hz, 3-H), 3.03 (d, 1H, J = 14.5 Hz, 4-CH2), 2.99 (m, 2H, 1'-H), 2.71 (dd, 1H , J = 13.5, 7.4 Hz, 3'-H), 2.64 (dd, 1H, J = 13.4, 6.6 Hz, 3'-H), 1.08 (d, 3H, J = 7.5 Hz, 3-CH3). 13C NMR (100 MHz, CDCI3): 5 171.3 (COO), 170.3 (C2),

156.3 (OCON), 137.5, 137.1, 135.1, 134.8, 129.8, 129.5, 129.45, 129.4, 129.2, 129.1, 128.95, 128.9, 128.85, 128.8, 128.6, 128.55, 128.5, 128.0, 127.6, 126.6 (Ar), 69.1 ( C4), 67.9, 66.4 (OCH2), 54.2 (C3), 51.6 (C2 '), 47.1 (C1'), 41.3 (4-CH2). 39.4 (C3 ’),

10.6 (3-CH3). MS (ES) +: 577.25 [M + H] +.

EXAMPLE 21

4S-Benzyloxycarboml-3S, 4S-dimetiM - [(2’R-bencMoxicarbomlammo-3’-feml) prop-

1’-il] -2-oxoazetidine

image83

Sirupe Rdto: 30% (from 7h, B: BTPP). Eluent: AcOEt: Hexane (1: 2). HPLC: t R = 17.49 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (400 MHz, CDCI3): 5 7.35- 7.15 (m, 15H, Ar), 5.36 (d, 1H, J = 8.4 Hz, 2-NH), 5.18 (d, 1H, J = 12.1 Hz, OCH2) , 5.12 (d, 1H, J = 12.1 Hz, OCH2), 5.03 (s, 2H, OCH2), 4.10 (m, 1H, 2'-H), 3.32 (dd, 1H, J = 14.6, 8.1 Hz, 1 '-H), 3.22 (dd, 1H, J = 14.6, 5.1 Hz, 1'-H), 2.96 (q, 1H, J = 7.5 Hz, 3-H), 2.87 (dd, 1H, J = 13.8, 6.2 Hz, 3'-H), 2.81 (m, 1H, 3'-H), 1.58 (s, 3H, CH3), 1.06 (d, 3H,

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J = 7.5 Hz, CH3). 13C NMR (75 MHz, CDCI3): 5 171.5 (COO), 170.0 (C2), 156.2 (OCON), 137.3, 136.8, 135.1, 129.5, 128.85, 128.8, 128.7, 128.5, 128.1, 126.7 (Ar), 67.5, 66.6 (OCH2), 64.8 (C4), 56.1 (C3), 51.7 (C2 '), 44.6 (C1'), 39.2 (C3 '), 20.8 (4- CH3), 10.4 (CH3). MS (ES) +: 501.37 [M + H] +. Exact mass calculated for C30H32N2O5: 500.23112; Found: 500.23117.

EXAMPLE 22

4R, S-BencM-4-bencMoxicarbomM - [(2’S-dibencilammo-3’-feml) prop-1’-M] -2-

oxoazetidine

image84

Sirupe Rdto: 10% (from 6m). Eluent: AcOEt: Hex (1: 4). Proportion of diastereoisomers, M / m = 5: 1. HPLC: t R = 7.55 min (gradient from 5% to 100% of A, in 20 min). 1 H NMR (400 MHz, CDCl 3): majority isomer 5 7.40-6.73 (m, 25H, Ar),

5.03 (d, 1H, J = 12.0 Hz, OCH2), 4.93 (d, 1H, J = 12.0 Hz, OCH2), 3.69 (d, 2H, J = 13.8 Hz, NCH2), 3.52 (d, 2H, J = 13.8 Hz, NCH2), 3.45 (m, 2H, 1'-H), 3.28 (d, 1H, J = 14.8 Hz, 3- H), 3.21 (m, 1H, 2'-H), 3.00 (d, 1H, J = 13.5 Hz, 4-CH2), 2.92 (dd, 1H, J = 14.2, 8.1 Hz, 3'-H), 2.82 (d, 1H, J = 14.8 Hz 3-H), 2.81 (m, 1H, 3'-H), 2.38 (d, 1H, J = 13.4 Hz, 4-CH2). 13C NMR (75 MHz, CDCI3): 5 170.0 (COO), 167.1 (C2), 140.2, 139.6, 135.0, 134.7, 129.55, 129.5, 129.1, 128.9, 128.8, 128.3, 127.4, 127.0, 126.1 (Ar), 67.6 (OCH2), 63.9 (C4), 58.5 (C2 '), 53.4 (NCH2), 45.3 (C3), 41.6 (C1'), 40.6 (4-CH2), 36.5 (C3 '). MS (ES) +: 608.88 [M + H] +.

EXAMPLE 23

4S-Benzyl -] - (4-benzyloxycarbonyl-3S-methyl-1 - [(2’S-dibenzylamino-3’-phenyl) prop-1’-

il] -2-oxoazetidine

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image85

Sirupe Rdto: 25% (from 7f). Eluent: AcOEt: Hex (1: 4). HPLC: t R = 7.19 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (400 MHz, CDCI3): 5 7.46-6.89 (m, 25H, Ar), 5.23 (d, 1H, J = 11.9 Hz, OCH2), 5.01 (d, 1H, J = 11.9 Hz, OCH2), 3.64 (d, 2H, J = 13.9 Hz, NCH2), 3.49 (m, 1H, 2'-H), 3.44 (d, 2H, J = 13.9 Hz, NCH2), 3.41 (dd, 1H, J = 13.9, 4.0 Hz, 1'-H), 3.22 (dd, 1H, J = 13.8, 9.8 Hz, 1'-H), 3.10 (q, 1H, J = 7.5 Hz, 3- H), 2.99 (d, 1H, J = 14.2 Hz, 4-CH2), 2.93 (d, 1H, J = 14.2 Hz, 4-CH2), 2.86 (dd, 1H, J = 14.4, 5.0 Hz, 3'-H), 2.75 (dd, 1H, J = 14.4, 9.1 Hz, 3'-H), 1.07 (d, 3H, J = 7.5 Hz, CH3). 13C NMR (75 MHz, CDCI3): 5 170.8 (COO), 179.1 (C2), 140.4, 139.7, 135.2, 135.0, 130.0, 129.6, 129.2, 128.9, 128.8, 128.7, 128.6, 128.2, 128.1, 127.3, 126.9, 125.9 (Ar),

68.4 (OCH2), 67.2 (C4), 57.9 (C2 ’), 57.8 (C3), 53.3 (NCH2), 43.0 (C1’), 40.9 (4-CH2), 36.2 (C3 ’), 10.6 (CH3). MS (ES) +: 623.01 [M + H] +.

EXAMPLE 24

4R-BencM-4-bencMoxicarboml-3R-metiM - [(2’S-dibencMammo-3’-feml) prop-1’-M] -

2-oxoazetidine

image86

Sirupe Rdto: 57% (from 7g). Eluent: AcOEt: Hex (1: 4). HPLC: t R = 7.29 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (400 MHz, CDCI3): 5 7.37-6.93 (m, 25H, Ar), 5.06 (d, 1H, J = 12.0 Hz, OCH2), 4.84 (d, 1H, J = 12.0 Hz, OCH2), 3.56 (d, 2H, J = 13.8 Hz, NCH2), 3.47 (d, 2H, J = 13.9 Hz, NCH2), 3.46 (m, 1H, 3-H), 3.37 (m, 1H, 2'-H), 3.14 (dd, 1H, J = 14.2, 8.1 Hz, 1'-H), 3.08 (m, 2H, 3'-H, 4-CH2), 2.81 (m, 3H, 4- CH2, 1'-H, 3'-H), 1.00 (d, 3H, J = 7.5 Hz, CH3). 13C NMR (75 MHz, CDCI3): 5 170.9 (COO), 169.9 (C2), 140.3, 140.0, 135.5, 135.1, 129.9, 129.7, 129.1, 129.0, 128.8,

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128.75, 128.7, 128.2, 127.5, 126.9, 126.0 (Ar), 68.6 (C4), 67.3 (OCH2), 58.7 (C2 '), 54.4 (C3), 53.2 (NCH2), 42.1 (C1'), 40.6 (4 -CH2), 35.9 (C3 '), 10.5 (CH3). MS (ES) +: 622.95 [M + H] +.

EXAMPLE 25

4R, S-BencM-4-methoxycarbomM - [(2’S-dibencMammo-3’-feml) prop-1’-M] -2-

oxoazetidine

image87

Sirupe Rdto: 71% (from 6n). Eluent: AcOEt: Hexane (1: 6). HPLC: t R = 6.49, 8.00 min (gradient from 30% to 95% of A in 10 min). Proportion of diastereoisomers, M: m = 1.8: 1. 1H NMR (400 MHz, CDCl3, majority diastereoisomer): 5 7.34-6.89 (m, 20H, Ar), 3.72 (m, 4H, NCH2), 3.62 (m, 1H, 1'-H), 3.45 (s, 3H , OCH3), 3.43 (dd, 1H, J = 14.3, 8.3 Hz, 1'-H), 3.27 (m, 1H, 2'-H), 3.25 (d, 1H, J = 14.8 Hz, 3-H) , 3.02 (d, 1H, J = 13.4 Hz, 4-CH2), 2.82 (d, 1H, J = 14.9 Hz, 3-H), 3.02 (m, 1H, 3'-H), 2.80 (m, 1H , 3'-H), 2.45 (d, 1H, J = 13.4 Hz, 4-CH2). 13C NMR (75 MHz, CDCI3): 5

170.6 (COO), 166.9 (CON), 140.2, 140.1, 139.9, 134.6, 129.5, 129.4, 129.2, 128.8, 128.7, 128.5, 128.3, 127.5, 127.0, 127.1 (Ar), 63.7 (C4), 58.8 (NCH2) , 53.4 (C2 '), 52.3 (OCH3), 45.1 (C3), 41.4 (C1'), 40.6 (4-CH2), 36.1 (C3 '). MS (ES) +: 533.27 (M + H +).

EXAMPLE 26

4R, S-Benzyl-4-ferc-butoxycarbonyl-1 - [(2’S-dibenzylamino-3-phenyl) prop-1’-il] -2-

oxoazetidine

image88

Sirupe Rdto: 41% (from 6th). Eluent: AcOEt: Hexane (1: 6). HPLC: t R = 7.28, 8.27 min (gradient from 50% to 95% of A in 10 min). Proportion of diastereoisomers, M: m = 1.8: 1. 1 H NMR (400 MHz, CDCl 3, diastereoisomer

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majority): 5 7.39-6.89 (m, 20H, Ar), 3.81 (d, 2H, J = 13.4, NCH2), 3.57 (m, 2H, 1'-H), 3.51 (d, 4H, J = 13.4, NCH2), 3.30 (m, 1H, 2'-H), 3.24 (d, 1H, J = 14.8, 3-H), 3.09 (d, 1H, J = 13.6, 4-CH2), 2.94 (m, 2H , 3'-H), 2.82 (d, 1H, J = 14.8, 3-H), 2.40 (d, 1H, J = 13.5 Hz, 4-CH2), 1.39 (s, 9H, CH3, fBu). 13C NMR (75 MHz, CDCI3): 5 169.3 (COO), 167.4 (CON), 140.0, 139.6, 135.2, 129.6, 129.5, 128.8, 128.5, 128.3, 128.2, 127.3, 127.1, 126.1 (Ar), 83.0 (C fBu), 64.2 (C4), 58.6 (C2 '), 53.5 (NCH2), 45.6 (C3), 41.8 (C1'), 40.9 (4-CH2), 36.8 (C3 '), 28.1 (CH3, fBu). MS (ES) +: 576.40 (M + H +).

EXAMPLE 27

4S-BencM-3S-metM-4-methoxycarbomM - [(2’S-dibencMammo-3’-feml) prop-1’-M] -2-

oxoazetidine

image89

Sirupe Rdto: 51% (from 7i). Eluent: AcOEt: Hexane (1: 8). HPLC: t R = 8.11 min (gradient of 15% to 95% of A in 10 min). 1H NMR (400 MHz, CDCI3): 5 7.30-7.02 (m, 20H, Ar), 3.73 (d, 2H, J = 14.2 Hz, NCH2), 3.69 (d, 2H, J = 14.2 Hz, NCH2), 3.59 (dd, 1H, J = 13.7, 5.1 Hz, 1'-H), 3.51 (m, 4H, 2'-H, OCH3), 3.19 (dd, 1H, J = 13.7, 7.6 Hz, 1'- H) , 3.10 (q, 1H, J = 7.5 Hz, 3-H), 3.04 (d, 2H, J = 13.6 Hz, 4-CH2), 2.99 (d, 2H, J = 13.6 Hz, 4-CH2), 2.97 (dd, 1H, J = 14.1, 7.7 Hz, 3'-H), 2.81 (dd, 1H, J = 14.1, 6.2 Hz, 3'-H), 1.14 (d, 3H, J = 7.5 Hz, 3- CH3). 13C NMR (75 MHz, CDCI3): 5 171.4 (COO), 169.9 (CON), 140.3, 139.9, 135.1, 130.0, 129.6, 128.8, 128.7, 128.2, 127.4, 126.9, 126.0 (Ar), 68.6 (C4), 58.7 (C2 '), 53.5 (NCH2), 53.2 (C3), 51.9 (OCH3), 42.7 (C1'), 40.6 (4- CH2), 36.0 (C3 '), 10.5 (3-CH3). MS (ES) +: 548.34 (M + H +).

EXAMPLE 28

4S-BencM-4-ferc-butoxicarboml-3S-metiM - [(2’S-dibencMammo-3’-feml) prop-1’-M] -

2-oxoazetidine

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image90

Sirupe Rdto: 77% (from 7j). Eluent: AcOEt: Hexane (1: 8). HPLC: t R = 2.33 min (gradient from 50% to 95% of A in 5 min). 1H NMR (400 MHz, CDCh): 5 7.31-7.04 (m, 20H, Ar), 3.76 (d, 2H, J = 13.9 Hz, NCH2), 3.56 (m, 4H, NCH2, 1'-H, 2 ' -H), 3.42 (m, 1H, 1'-H), 3.14 (q, 1H, J = 7.5 Hz, 3-H), 2.95 (m, 4H, 3'-H, 4-CH2), 1.46 ( s, 9H, CH3 fBu), 1.19 (d, 3H, J = 7.5 Hz, 3-CH3). 13C NMR (75 MHz, CDCI3): 5 170.2 (COO), 169.7 (CON), 140.2, 139.6, 135.6, 129.8, 129.5, 128.6, 128.3, 128.1, 127.9, 127.0, 126.8,

125.7 (Ar), 82.8 (C fBu), 68.3 (C4), 58.0 (C2 '), 53.3 (NCH2), 53.1 (C3), 42.7 (C1'), 41.3 (4-CH2), 36.4 (C3 ') , 28.0 (CH3 fBu), 10.7 (3-CH3). MS (ES) +: 590.20 (M + H +).

Hydrogenolysis debenzylation of the p-lactam of Example 28.

16 (0.17 mmol, 0.10 g) is dissolved in 20 mL of methanol and the solution is cooled to 0 ° C. A 20% ratio of Pd (OH) 2 (0.02 g) is added to the mixture, together with HCl (1 eq). The mixture is hydrogenated at 40 psi and at room temperature for 22h to try to completely debencylate. The catalyst is filtered and the solvent is evaporated under reduced pressure. Wash with AcOEt and a 10% solution of NaHCO3. The organic phase is dried over Na2SO4 and evaporated under reduced pressure. The residue is purified using silica gel column chromatography.

EXAMPLE 29

4S-Benzyl-4-ferc-butoxicarboml-3S-metiM - [(2’S-benzylamm-3’-feml) prop-1’-il] -2-

oxoazetidine

image91

Sirupe Rdto: 29% (from the compound of Example 28). Eluent: AcOEt: Hexane (1: 2). HPLC: t R = 3.62 min (gradient of 15% to 95% of A in 5 min). 1H NMR (400 MHz, CDCl3): 5 7.32-7.12 (m, 15H, Ar), 3.79 (d, 1H, J = 13.1 Hz, NCH2), 3.73 (d, 1H,

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J = 13.1 Hz, NCH2), 3.43 (d, 1H, J = 14.5 Hz, 4-CH2), 3.29 (m, 1H, 2'-H), 3.16 (m, 1H, 1'- H), 3.09 ( m, 3H, 3-H, 1'-H, 4-CH2), 2.80 (dd, 1H, J = 13.9, 6.4 Hz, 3'-H), 2.73 (dd, 1H, J = 13.9, 6.7 Hz, 3'-H), 1.39 (s, 9H, CH3, fBu), 1.21 (d, 3H, J = 7.5 Hz, 3-CH3). 13C NMR (75 MHz, CDCI3): 5 170.5 (COO), 170.1 (CON), 140.6, 138.9, 135.6, 130.0, 129.5, 128.6, 128.5, 128.4, 128.3, 127.3, 126.9, 126.3 (C, Ar), 83.2 (C fBu), 68.5 (C4), 57.6 (C2 '), 53.4 (C3), 51.4 (NCH2), 47.5 (C1'), 41.0 (4-CH2), 39.5 (C3 '), 28.1 (CH3, fBu ), 10.7 (3-CH3). MS (ES) +: 499.07 (M + H +).

EXAMPLE 30

4S-BencM-4-ferc-butoxicarboml-3S-metiM - [(2’S-ammo-3’-feml) prop-1-M] -2-

oxoazetidine

image92

Sirupe Rdto: 29% (from the compound of Example 28). Eluent: MeOH: AcOEt (1: 3). HPLC: t R = 3.22 min (gradient of 15% to 95% of A in 5 min). 1H NMR (400 MHz, CDCh): 5 7.29-7.09 (m, 10H, Ar), 3.47 (d, 1H, J = 14.6 Hz, 4-CH2), 3.42 (m, 1H, 2'-H), 3.10 (d, 1H, J = 14.6 Hz, 4-CH2), 3.08 (q, 1H, J = 7.6 Hz, 3-H), 3.02 (m, 2H, 1'-H), 2.66 (dd, 1H, J = 13.5, 5.7 Hz, 3'-H), 2.53 (dd, 1H, J = 13.5, 8.1 Hz, 3'-H), 1.47 (s, 9H, CH3 fBu), 1.21 (d, 3H, J = 7.5 Hz, 3-CH3). 13C NMR (75 MHz, CDCh): 5 170.5 (COO),

170.4 (CON), 138.7, 135.5, 130.1, 129.4, 128.7, 128.6, 127.5, 126.5 (Ar), 83.4 (C fBu), 68.6 (C4), 53.6 (C3), 51.5 (C2 '), 50.1 (C1' ), 42.1 (C3 '), 40.7 (4-CH2), 28.3 (CH3, fBu), 10.8 (3-CH3). MS (ES) +: 409.09 (M + H +).

EXAMPLE 31

4S-Benzyl-4-ferc-butoxycarbonyl-3S-methyl-1- [2’S-benzyloxycarbonylamino-3’-

phenyl) prop-1’-il] -2-oxoazetidine

image93

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30

The compound of Example 30 (0.19 mmol, 0.08 g) is dissolved in anhydrous dichloromethane and TEA (0.19 mmol, 0.02 g) is added. The solution is cooled to 0 ° C and propylene oxide (2.90 mmol, 0.17 g) is added, followed by benzyl chloroformate (0.39 mmol, 0.02 g). The reaction proceeds at room temperature for 1h 40min. The solvent is evaporated under reduced pressure, the residue is dissolved in AcOEt and washed with water and a saturated NaCl solution. The organic phase is dried over Na2SO4 and evaporated under reduced pressure. The residue is purified using silica gel column chromatography. Sirupe Rdto: 69%. Eluent: Hexane (1: 4). HPLC: t R = 9.05 min (gradient from 30% to 95% of A in 10 min). 1 H NMR (400 MHz, CDCh): 5 7.36

7.04 (m, 15H, Ar), 6.00 (d, 1H, J = 8.3 Hz, NH), 4.98 (s, 2H, OCH2), 4.11 (m, 1H, 2'-H), 3.35 (d, 1H, J = 14.5 Hz, 4-CH2), 3.16 (dd, 1H, J = 14.5, 6.9 Hz, 1'-H), 3.07 (m, 2H, 1'-H,

3-H), 2.99 (d, 1H, J = 14.5 Hz, 4-CH2), 2.87 (dd, 1H, J = 13.9, 7.7 Hz, 3'-H), 2.75 (dd, 1H, J = 14.1, 6.9 Hz, 3'-H), 1.40 (s, 9H, CH3 fBu), 1.15 (d, 3H, J = 7.5 Hz, 3-CH3). 13C NMR (75 MHz, CDCl3): 5 170.9 (COO), 170.4 (CON), 156.1 (NCOO), 138.0, 137.0, 135.2, 129.9, 129.4, 128.5, 128.4, 127.9, 127.5, 126.5 (Ar), 83.7 ( C fBu), 68.6 (C4), 66.3 (OCH2), 53.5 (C3), 51.8 (C2 '), 46.6 (C1'), 40.7 (4-CH2), 38.9 (C3 '), 28.2 (CH3 fBu), 10.8 (3-CH3). MS (ES) +: 543.16 (M + H +).

EXAMPLE 32

4S-Benzyl-4-ferc-butoxycarbonyl-3S-methyl-1 - [(2’S-ferc-butoxycarbonylamino-3-

phenyl) prop-1’-il] -4-oxoazetidine

image94

Di-tert-butyl dicarbonate (0.08 mmol, 0.02 g) is added to a solution of the compound of Example 30 (0.05 mmol, 0.02 g) in anhydrous dichloromethane. The reaction mixture is allowed to stir for 24 hours at room temperature, evaporated under reduced pressure and purified by silica gel flash column chromatography. Sirupe Rdto: 38%. Eluent: AcOEt: Hexane (1: 4). HPLC: t R = 9.31 min (gradient from 50% to 95% of A in 10 min). 1H NMR (300 MHz, CDCh): 5 7.28-7.10 (m, 10H, Ar), 5.62 (d, 1H, J = 7.7 Hz, NH), 4.13 (m, 1H, 2'-H), 3.38 (d , 1H, J = 14.3 Hz, 4-CH2), 3.15 (m, 5H, 3- H, 1'-H, 4-CH2), 2.90 (m, 1H, 3'-H), 2.80 (dd, 1H , J = 13.9, 6.7 Hz, 3'-H), 1.47 (s, 9H, CH3 OfBu), 1.36 (s, 9H, 3-CH3, Boc), 1.21 (d, 3H, J = 7.5 Hz, 3- CH3). 13C NMR (75

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MHz, CDCI3): 5 170.8 (COO), 170.4 (CON), 155.6 (NCOO), 138.3, 135.3, 130.0,

129.5, 128.7, 128.5, 127.5, 126.4 (Ar), 83.5 (C, OfBu), 79.0 (C, Boc), 68.6 (C4), 53.3 (C3), 51.1 (C2 '), 46.6 (C1'), 40.6 (4-CH2), 39.0 (C3 '), 28.5 (CH3, OfBu), 28.3 (CH3, Boc), 10.8 (3-CH3). MS (ES) +: 509.36 (M + H +).

Synthesis of p-lactams dicarboxylate

A solution of the corresponding substituted 4-benzyloxycarbonyl derivative (0.2 mmol) in MeOH (17 mL) is hydrogenated at room temperature and 15 psi of pressure for 7 h, using Pd-C (10%) as catalyst. Once the catalyst has been filtered off, the solvent is evaporated to dryness.

EXAMPLE 33

4R, S-Bencil-4-carboxM- [3’S-tert-butoxycarbomlammo-3’-carboxy) prop-1’-il] -2-

oxoazetidine

image95

Sirupe Rdto: 99% (from the compound of Example 3). Proportion of diastereoisomers, M / m = 6: 1. HPLC: t R = 9.17 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (400 MHz, CDCl3, majority diastereoisomer): 5 9.40 (s, 2H, OH), 7.38-7.06 (m, 5H, Ph), 5.68 (d, 1H, J = 8.5 Hz, NH), 4.24 (m , 1H, 3'-H), 3.54-2.73 (m, 6H, 1'-H, 3-H, 4-CH2), 2.17 (m, 2H, 2'-H), 1.43 (s, 9H, fBu ). 13C NMR (CDCl3): 5 175.7, 174.6 (COO), 168.3 (C2), 156.3 (OCON), 134.1, 130.0, 128.9, 127.7, (Ar), 80.8 (C fBu), 63.4 (C4), 51.8 (C3 '), 44.8 (C3), 38.8 (C1', 4-CH2), 30.6 (C2 '), 28.4 (CH3 fBu). MS (ES) +: 407.20 [M + H] +.

EXAMPLE 34

4R, S-BenciM- [4’S-tert-butoxycarbomlammo-4’-carboxy) but-1’-il] -4-carboxy-2-

oxoazetidine

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30

image96

Sirupe Rdto: 74% (from the compound of Example 12). Proportion of diastereoisomers, M / m = 3: 2. HPLC: t R = 9.46 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (300 MHz, CDCl3, Major Diastereoisomer): 5 10.09 (s, 2H, OH), 7.24-7.07 (m, 5H, Ar), 5.41 (d, 1H, J = 8.1 Hz, NH), 4.18 (m , 1H, 4'-H), 3.50-2.62 (m, 6H, 1'-H, 3-H, 4-CH2), 1.88-1.56 (m, 4H, 3'-H, 2'-H), 1.36 (s, 9H, fBu). 13C NMR (CDCl3, Major Diastereoisomer): 5 176.2, 174.5 (COO), 168.1 (C2), 156.1 (OCON), 134.6, 129.9, 128.8, 127.5 (Ar), 80.6 (C fBu), 63.1 (C4), 53.1 (C4 '), 45.1 (C3), 41.9 (C1'), 38.9 (4-CH2), 29.9 (C3 '), 28.4 (CH3 fBu), 24.0 (C2'). MS (ES) +: 421.26 [M + H] +.

Synthesis of p-lactams amldicas

Method A. A solution of the corresponding substituted 4-benzyloxycarbonyl derivative (0.2 mmol) in MeOH (17 mL) is added 20% by weight of Pd-C and hydrogenated at room temperature and 30 psi for 3h. The corresponding carboxylated derivative (0.143 mmol) is then dissolved in dry CH2Cl2 (4 mL) and the desired amine (0.286 mmol), PyBOP (benzotriazol-1-yl-N-oxy-tris hexafluorophosphate is added consecutively. pyrrolidin) phosphonium) (0.286 mmol, 149 mg) and TEA (0.286 mmol, 40 ^ L). After 24 h of stirring at room temperature the solvent is evaporated to dryness. The crude obtained is dissolved in AcOEt and washed successively with 10% dichloric acid solution, 10% NaHCO3 solution, H2O and saturated NaCl solution. The organic phase is dried over Na2SO4 and evaporated, the resulting residue being purified by silica gel column chromatography using the eluent system indicated in each case.

Method B. To a solution of the corresponding substituted carboxy derivative (0.155 mmol) in dry CH2Cl2 (2 mL) or anhydrous DMF (2 mL) the desired amine (0.62 mmol), PyBOP (0.62) is added consecutively mmol) and TEA (0.62 mmol). After stirring for 24 hours at room temperature, the solvent is evaporated to dryness. The crude obtained is dissolved in AcOEt and washed with 0.1N HCl solution. The aqueous solution is basified with 1M NaOH and extracted with AcOEt. The phase

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The organic was washed with saturated NaCl solution, dried over anhydrous Na2SO4 and evaporated, the resulting residue was purified by silica gel column chromatography, using the eluent system indicated in each case.

EXAMPLE 35

4S-Benzyl-4 - [(N-benzyl) carbamoyl] -1 - [(3’S-ferc-butoxycarbonylamino-3 ’- [(N-

benzyl) carbamoyl] prop-1’-il] -2-oxoazetidine

image97

Sirupe Rdto: 80% (from the compound of Example 33, Method A). Eluent: AcOEt: Hexane (2: 1). HPLC: t R = 13.79 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (400 MHz, CDCfe): 5 7.62 (sa, 1H, NHCO), 7.31-7.12 (m, 15H, Ar), 6.11 (sa, 1H, NHCO), 5.33 (sa, 1H, NHCO), 4.44 ( m, 2H, NCH2), 4.40 (dd, 1H, J = 14.4, 5.7 Hz, NCH2), 4.30 (dd, 1H, J = 14.4, 5.7 Hz, NCH2Ph), 4.23 (m, 1H, 3'-H) , 3.47 (m, 1H, 1'-H), 3.39 (d, 1H, J = 14.0 Hz, 4-CH2), 3.32 (m, 2H, 1'-H, 4-CH2), 3.04 (d, 1H , J = 14.6 Hz, 3- H), 2.95 (d, 1H, J = 14.6 Hz, 3-H), 2.05 (m, 1H, 2'-H), 1.94 (m, 1H, 2'-H) , 1.42 (s, 9H, CH3 fBu). 13C NMR (75 MHz, CDCh): 171.5 (CON), 170.8 (CON), 167.4 (C2), 155.5 (OCON), 138.1, 137.4, 129.8, 128.95, 128.9, 128.8, 128.3, 127.95, 127.9, 127.6 (Ar ), 81.0 (C fBu), 64.2 (C4), 52.4 (C3 '), 46.4 (C3), 44.0, 43.7 (NHCH2), 40.6 (C1'), 39.3 (4- CJ), 29.9 (C2 '), 28.5 (CH3 fBu). MS (ES) +: 585.55 [M + H] +. Exact mass calculated for C34H40N4O5: 584.29987; Found: 584.30012.

EXAMPLE 36

4R, S-BenciM- [4’S-tert-butoxycarbomlammo-4 ’- [(N-bencM) carbamoM] but-1’-M] -4-

[(N-benzyl) carbamoyl] -2-oxoazetidine

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Sirupe Rdto: 27% (from the compound of Example 34, Method B). Eluent: AcOEt: Hex (4: 1). Proportion of diastereoisomers, M / m = 3: 2. HPLC: t R = 13.80 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (400 MHz, CDCI3, Major Diastereoisomer): 5 7.27-6.88 (m, 15H, Ar), 5.95 (m, 1H, CONH), 5.32 (m, 1H, CONH), 4.52-4.06 (m, 5H, NCH2, 4'-NH), 3.58 (m, 1H, 4'-H), 3.51-2.86 (m, 6H, 1'-H, 3-H, 4-CH2), 1.91-1.62 (m, 4H, 3'-H, 2'-H), 1.35 (s, 9H, CH3 fBu). 13C NMR (75 MHz, CDCl3, Major Diastereoisomer): 5 172.3, 171.0 (CONH), 167.2 (C2), 156.1 (OCON), 138.3, 137.3, 135.0, 129.9, 129.8, 128.9, 128.8, 128.7, 128.1, 128.0, 127.7,

127.5, (Ar), 80.0 (C fBu), 63.8 (C4), 52.5 (C4 '), 46.2 (C3), 43.9, 43.5 (NHCH2), 41.1 (C1'), 39.6 (4-CH2), 30.9 ( C3 '), 28.5 (CH3 fBu), 24.6 (C2'). MS (ES) +: 599.43 [M + H] +.

EXAMPLE 37

4S-4-Benzyl-4 - [(N-benzyl) carbamoyl] -3S-methyl-1- [3’S-ferc-butoxycarbonylamino-3’-

[(N-benzyl) carbamoyl] prop-1’-yl] -2-oxoazetidine

image99

Sirupe Rdto: 55% (from the compound of Example 6, Method A). Eluent: Acetone: DCM (1:25). HPLC: t R = 14.18 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (400 MHz, CDCh): 5 8.32 (m, 1H, NHCO), 8.20 (m, 1H, NHCO), 7.45-7.19 (m, 15H, Ar), 5.99 (d, 1H, J = 6.6 Hz, 3'-NH), 4.53 (m, 5H, NCH2, 3'-H), 3.69 (d, 1H, J = 14.9 Hz, 4-CH2), 3.35 (m, 2H, 1'-H, 4-CH2 ), 3.10 (m, 1H, 1'-H), 3.04 (q, 1H, J = 7.5 Hz, 3-H), 2.30 (m, 1H, 2'-H), 1.94 (m, 1H, 2 ' -H), 1.39 (s, 9H, CH3 fBu), 1.01 (d, 3H, J = 7.5 Hz, 3-CH3) .13C NMR (75 MHz, CDCh): 173.2 (CON), 171.0 (CON), 170.0 (CON), 156.5 (OCON), 138.9, 138.2, 135.7, 130.2, 128.9, 128.7, 128.4, 128.2, 127.7, 127.4, 127.3, 127.1 (Ar), 80.2 (C fBu), 69.1 (C4), 53.5 (C3 ), 51.9 (C3 '), 43.8, 43.6 (NCH2),

41.4 (C1 ’), 38.6 (4-CH2), 34.1 (C2’), 28.4 (CH3 fBu), 9.6 (CH3). MS (ES) +: 599.31 [M + H] +. Exact mass calculated for C35H42N4O5: 598.31552; Found: 598.31608.

EXAMPLE 38

4R-4-BencM-4 - [(N-bencM) carbamoil] -3R-metiM- [3’S-feA'c-butoxycarbomlammo-3’-

[(N-benzyl) carbamoyl] prop-1’-yl] -2-oxoazetidine

image100

Sirupe Rdto: 37% (from the compound of Example 6, Method A). Eluent: Acetone: DCM (1:15). HPLC: t R = 14.40 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (400 MHz, CDCI3): 5 7.96 (sa, 1H, NHCO), 7.30-7.20 (m, 15H, Ar), 6.81 (sa, 5 1H, NHCO), 5.17 (d, 1H, J = 6.6 Hz , 3'-NH), 4.58 (dd, 1H, J = 14.9, 6.1 Hz, NCH2), 4.35

(m, 3H, J = 14.9, 6.1 Hz, NCH2), 4.14 (m, 1H, 3'-H), 3.76 (d, 1H, J = 14.7 Hz, 4-CH2), 3.11 (m, 4H, 1 '-H, 4-CH2, 3-H), 1.84 (m, 2H, 2'-H), 1.40 (s, 9H, CH3 fBu), 1.10 (d, 3H, J = 7.5 Hz, CH3). 3C NMR (75 MHz, CDCI3): 171.9 (CON), 170.0 (CON), 155.5 (OCON), 138.0, 136.0, 130.2, 129.9, 128.9, 128.8, 128.7, 128.4, 128.3, 127.8, 127.6, 127.4 (Ar) , 10 78.8 (C fBu), 69.7 (C4), 54.6 (C3), 52.7 (C3 '), 44.0, 43.6 (NCH2), 40.7 (4-CH2), 40.1

(C1 ’), 32.1 (C2’), 28.4 (CH3 fBu), 10.2 (3-CH3). MS (ES) +: 599.31 [M + H] +. Exact mass calculated for C35H42N4O5: 598.31552; Found: 598.31608.

EXAMPLE 39

15 4S-BencM-4- [N - [(pmdm-4 ’’ - M) metM] carbamoiM - [(3’S-ferc-butoxicarbomlammo-3’-

[N - [(pyridin-4 ’- il) methyl] carbamoil] prop-1’-il] -2-oxoazetidine

image101

(19a) Sirupe. Rdto: 37% (from the compound of Example 33, Method B). Eluent: CH2Cl2: MeOH (9: 1). HPL: t R = 1.67 min (gradient of 5% to 100% of A, in 20 min). 1H 20 NMR (400 MHz, DMSO-da): 5 8.65 (t, 1H, J = 5.9 Hz, CONH), 8.46 (t, 1H, J = 5.9 Hz, CONH), 8.43 and 7.31-7.04 (m, 14H , Ar, 3'-NH), 4.36-4.22 (m, 4H, NCH2,), 3.94 (m, 2H, 3'-H), 3.42 (d, 1H, J = 13.7 Hz, 4-CH2), 3.30 -3.18 (m, 2H, 1'-H), 3.13 (d, 1H, J = 13.7 Hz, 4-CH2), 3.12 (d, 1H, J = 14.7 Hz, 4-CH2), 2.89 (d, 1H , J = 14.7 Hz, 4-CH2), 2.07 (m, 1H, 2'-H), 1.88 (m, 1H, 2'-H), 1.39 (s, 9H, CH3 fBu). 13C NMR (75 Mz, DMSO-da): 5

173.1, 170.1 (CONH), 167.0 (C2), 156.3 (OCON), 149.8, 149.7, 149.4, 148.9, 135.6,

130.5, 129.0, 127.8, 122.9, 122.6 (Ar), 79.5 (C Bu), 63.9 (C4), 53.6 (C3), 45.1 (C3 '), 42.2, 41.7 (NCH2), 39.5 (C1', 4-CH2 ), 30.9 (C2 '), 28.7 (CH3 Bu). MS (ES) +: 587.40 [M + H] +.

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EXAMPLE 40

4R-Benzyl-4- [N - [(pyridin-4 ’’ - il) methyl] carbamoyl] -1- [3’S-ferc-butoxycarbonylamino-3’-

[N - [(pyridin-4 ’- il) methyl] carbamoil] prop-1’-il] -2-oxoazetidine

image102

10 (19b) Sirupe. Rdto: 20% (from the compound of Example 33, Method B). Eluent:

CH2Cl2: 10% MeOH. HPLC: t R = 1.67 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (400 MHz, CDCI3): 5 8.46 and 7.30-6.97 (m, 13H, Ar), 7.76 (m, 1H, CONH), 7.10 (m, 1H, CONH), 5.43 (d, 2H, J = 7.9 Hz, 3'-NH), 4.48-4.21 (m, 4H, NCH2), 3.57-3.17 (m, 7H, 1'-H, 3'-H, 3-H, 4-CH2), 1.50 (m, 2H, 2'-H), 1.41 (s, 9H, Bu). 13C NMR (CDCI3): 5 15 172.2, 171.3 (CONH), 167.5 (C2), 155.7 (OCON), 149.9, 149.8, 147.5, 146.9, 135.2,

129.8, 129.0, 127.7, 122.8, 122.4 (Ar), 80.4 (C Bu), 64.2 (C4), 52.6 (C3), 45.6 (C3 '), 42.7, 42.4 (NCH2), 40.2 (4-CH2), 39.5 (C1 '), 32.4 (C3'), 28.4 (CH3 Bu). MS (ES) +: 587.33 [M + H] +.

20 EXAMPLE 41

4R, S-Benzyl-4 - [(N-pyridin-3-yl) carbamoyl] -1 - [(3’S-ferc-butoxycarbonylamino-3 ’- [(N-

pyridin-3-yl) carbamoyl] prop-1’-yl] -2-oxoazetidine

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Sirupe Rdto: 26% (from the compound of Example 33, Method B). Eluent: CH2Cl2: 10% MeOH. Proportion of diastereoisomers, M / m = 5: 2. HPLC: t R = 3.27 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (400 MHz, CDCI3, Major Diastereoisomer): 5 9.60 (s, 1H, CONH), 9.13 (s, 1H, CONH), 8.74 (d, 1H, J = 2.5 Hz, 2-Py), 8.62 (d , 1H, J = 2.7 Hz, 2-Py), 8.36 (dd, 1H, J = 4.8, 1.5 Hz, Py), 8.32.7.29 (m, 5H, Py), 7.33-7.14 (m, 5H, Ph) , 5.82 (d, 1H, J = 7.6 Hz, 3'-NH), 4.68 (m, 1H, 3'-H), 3.58-3.18 (m, 4H, 4-CH2, 1'-H), 3.10 ( d, 1H, J = 15.2 Hz, 3-H), 3.00 (d, 1H, J = 15.0 Hz, 3-H), 2.39 (m, 1H, 2'-H), 1.99 (m, 1H, 2 ' -H), 1.17 (s, 9H, CH3 fBu). 13C NMR (300 MHz, CDCI3): 5 170.3, 169.0 (CONH), 167.6 (C2), 156.5 (OCON), 146.4, 145.9, 144.7, 141.4, 134.7, 133.9, 130.6, 130.2, 129.9, 129.2, 127.9, 126.9 , 123.8, 123.4 (Ar), 80.8 (C fBu), 63.9 (C4), 52.6 (C3 '), 47.2 (C3), 41.8 (C1'), 37.3 (4-CH2), 33.8 (C2 '), 28.13 (CH3 fBu). MS (ES) +: 559.40 [M + H] +.

EXAMPLE 42

4S-BencM-4 - [(N-pmdm-4-M) carbamoN] -1- [3’S-tert-butoxycarbomlammo-3 ’- [(N-

pyridin-4-yl) carbamoyl] prop-1’-yl] -2-oxoazetidine

image104

(23a) Sirupe. Rdto: 21% (from the compound of Example 33, Method B). Eluent: CH2Cl2: 10% MeOH. HPLC: t R = 3.26 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (400 MHz, DMSO-da): 5 10.41 (s, 1H, CONH), 10.20 (s, 1H, CONH), 8.44 (m, 8H, Py), 7.61-7.20 (m, 6H, Ph, 3 '-NH), 4.10 (m, 1H, 3'-H), 3.42 (d, 1H, J = 14.0 Hz, 4-CH2), 3.32 (d, 1H, J = 14.0 Hz, 4-CH2), 3.22 (d, 1H, J = 14.8 Hz, 3-H), 3.01 (m, 2H, 1'-H), 2.98 (d, 1H, J = 14.8 Hz, 3-H), 2.24 (m, 1H, 2 '-H), 2.06 (m, 1H, 2'-H), 1.39 (s, 9H, CH3 fBu). 13C NMR (75 MHz, CD3OD): 5 173.6, 172.7 (CONH), 169.0 (C2), 157.9 (OCON), 150.8, 150.7, 147.8, 147.5, 135.6, 131.3, 129.7, 128.6, 115.7, 115.2 (Ar), 80.8 (C Bu), 66.4 (C4), 55.1 (C3 '), 47.3 (C1'), 45.4 (C3), 40.5 (4-CH2), 32.1 (C2 '), 28.7 (CH3 fBu). MS (ES) +: 559.40 [M + H] +.

EXAMPLE 43

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4R-Bencil-4 - [(N- (pmdm-4-il) carbamoil] -1- [3’S-tert-butoxycarbomlammo-3 ’- [(N-

pyridin-4-yl) carbamoyl] prop-1’-yl] -2-oxoazetidine

image105

(23b) Sirupe. Rdto: 23% (from the compound of Example 33, Method B). Eluent: CH2Cl2: 10% MeOH. HPLC: t R = 3.26 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (400 MHz, DMSO-d6): 5 10.42 (s, 1H, CONH), 10.26 (s, 1H, CONH), 8.49 (m, 4H, Py), 7.65 (m, 2H, Py), 7.62 ( m, 2H, Py), 7.36-7.26 (m, 6H, Ph, 3'-NH), 4.18 (m, 1H, 3'-H), 3.47 (d, 1H, J = 14.1 Hz, 4-CH2) , 3.37 (d, 1H, J = 14.1 Hz, 4-CH2), 3.27 (d, 1H, J = 14.8 Hz, 3-H), 3.08 (m, 2H, 1'-H), 2.96 (d, 1H , J = 14.8 Hz, 3-H), 2.15 (m, 2H, 2'-H), 1.45 (s, 9H, CH3 fBu). 13C NMR (75 MHz, CD3OD): 5 173.7, 172.9 (CONH), 168.7 (C2), 158.0 (OCON), 150.8, 150.7, 147.9, 147.5, 131.7, 131.2, 129.7, 128.6, 115.9, 115.2 (Ar), 80.9 (C fBu), 66.2 (C4), 54.9 (C3 '), 45.3 (C3), 40.7 (C3'), 40.5 (4-CH2), 31.8 (C2 '), 28.7 (CH3 fBu). MS (ES) +: 559.40 [M + H] +.

Synthesis of dipeptldic derivatives

A solution of the corresponding substituted dibenzyloxycarbonyl derivative (0.2 mmol) in MeOH (17 mL) is hydrogenated at room temperature and 15 psi of pressure for 7 h, using Pd-C (10%) as catalyst. Once the catalyst has been filtered off, the solvent is evaporated to dryness. Subsequently, the corresponding substituted azetidinone dicarboxy (0.33 mmol) is dissolved in dry THF (4 mL) and HL-Ala-OMeHCl or HL-Phe-OMeHCl (0.66 mmol), PyBOP (0) are added consecutively , 66 mmol, 336 mg) and TEA (1.32 mmol, 0.18 mL). After 24-48 h of stirring at room temperature, it is analyzed by HPLC in order to determine the proportion of the diastereoisomers formed. The solvent is then evaporated to dryness. The crude obtained is dissolved in AcOEt and washed with 10% dichloric acid solution, 10% NaHCO3 solution, H2O and saturated NaCl solution. The organic phase is dried over Na2SO4 and evaporated, the resulting residue being purified by silica gel column chromatography using the eluent system indicated in each case.

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EXAMPLE 44

4S-Bencil-1 - [3’S-ferc-butoxicarbomlammo-3 ’- [W - [(1’’S-

methoxycarboml) etM] carbamoM] prop-1’-M] -4- [W - [(1 ’” S-

methoxycarbonyl) ethyl] carbamoyl] -2-oxoazetidine

image106

Sirupe Rdto: 28% (from the compound of Example 3). Eluent: MeOH: DCM (1:30). HPLC: t R = 4.30 min (gradient of 15% to 95% of A, in 5 min). 1H NMR (400 MHz, CDCI3): 5 8.38 (d, 1H, J = 7.3 Hz, 1 '' - NH), 7.53 (d, 1H, J = 7.5 Hz, 1 '' - NH), 7.317.18 (m, 15H, Ar), 5.67 (d, 1H, J = 7.4 Hz, 3'-NH), 4.70 (q, 1H, J = 7.5 Hz, 1 '' - H), 4.52 (q, 1H, J = 7.3 Hz, 1 '' - H), 4.34 (m, 1H, 3'-H), 3.72 (s, 4H, OMe, 1'-H), 3.69 (s, 3H, OMe), 3.48 (d , 1H, J = 14.8 Hz, 4-CH2), 3.38 (d, 1H, J = 14.8 Hz, 4-CH2), 3.18 (m, 1H, 1'-H), 2.98 (d, 1H, J = 15.1 Hz, 3-H), 2.90 (d, 1H, J = 15.1 Hz, 3-H), 2.22 (m, 1H, 2'-H), 2.04 (m, 1H, 2'-H), 1.52 (d , 3H, J = 7.2 Hz, 2 '' - H), 1.38 (d, 3H, J = 7.3 Hz, 2 '' - H), 1.37 (s, 9H, CH3 * Bu). 13C NMR (75 MHz, CDCI3): 173.4 (COO), 172.9 (COO), 171.1 (CON ''), 170.8 (CON ''), 169.6 (C2), 155.7 (OCON), 135.0, 130.4, 128.9, 127.5 (Ar), 79.9 (C, fBu), 63.6 (C4), 52.6 (OMe), 52.55 (OMe), 52.5 (C3 '), 48.7 (C1' '), 48.3 (C1' '), 46.9 ( C3), 40.3 (C1 '), 37.2 (4-CH2), 33.6 (C2'), 28.5 (CH3 * Bu), 17.7 (C2 ''), 17.1 (C2 '' '). MS (ES) +: 577.47 [M + H] +. Exact mass calculated for C28H40N4O9: 576.27953; Found: 576.27722.

EXAMPLE 45

4R-Bencil-1 - [3’S-ferc-butoxycarbonylamino-3 ’- [W - [(1’’S-

methoxycarbonyl) ethyl] carbamoyl] prop-1’-il] -4- [W - [(1 ’’ ’S-

methoxycarbonyl) ethyl] carbamoyl] -2-oxoazetidine

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image107

Sirupe Rdto: 12% (from the compound of Example 3). Eluent: MeOH: DCM (1:30). HPLC: t R = 4.28 min (gradient of 15% to 95% of A, in 5 min). Proportion of diastereoisomers M, m = 1: 0.7. 1H NMR (400 MHz, CDCl3, majority diastereoisomer): 5 7.62 (sa, 1H, NHCO), 7.31-7.15 (m, 15H, Ar), 7.04 (sa, 1H, NHCO), 5.43 (d, 1H, J = 6.6 Hz, 3'-NH), 4.52 (m, 2H, 1 '' - H, 1 '' '- H), 4.34 (m, 1H, 3'-H), 3.73 (s, 3H, OMe), 3.71 (s, 3H, OMe), 3.52 (m, 1H, 1'-H), 3.38 (m, 2H, 4-CH2), 3.25 (m, 1H, 1'- H), 3.17 (m, 1H, 1'-H), 3.02 (d, 1H, J = 14.7 Hz, 3-H), 2.97 (d, 1H, J = 14.7 Hz, 3-H), 2.05 (m, 1H, 2'-H), 1.84 (m, 1H, 2'-H), 1.41 (s, 9H, CH3 * Bu), 1.40 (d, 3H, J = 6.8 Hz, CH3), 1.32 (d, 3H, J = 7.2 Hz, CH3) . 13C NMR (75 MHz, CDCI3): 173.6 (COO), 173.1 (COO), 171.2 (CON), 171.1 (CON), 167.4 (C2), 155.7 (OCON), 134.9, 130.0, 128.9, 127.5 (Ar), 80.2 (C, fBu), 63.6 (C4), 52.8 (OMe), 52.5 (OMe), 52.4 (C3 '), 48.5 (C1' '), 48.3 (C1' '), C1' '), 47.1 (C3), 39.1 (C1 '), 37.1 (4-CH2), 32.1 (C2'), 28.4 (CH3 fBu), 18.1 (C2 ''), 17.6 (C2 '' '). MS (ES) +: 577.47 [M + H] +.

EXAMPLE 46

4R-BenciM- [3’S-ferc-butoxycarbomlammo-3 ’- [W - [(1” S-methoxycarboml-2’-

feml) etM] carbamoM] prop-1’-M] -4- [W - [(1 ’” S-methoxycarboml-2’feml) etM] carbamoil] -2-

oxoazetidine

image108

Sirupe Rdto: 12% (from the compound of Example 3). Semi-preparative HPLC purification (gradient from 50% to 60% of A, in 30 min). HPLC: t R = 15.13 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (400 MHz, CDCh): 5 7.49 (m, 2H, NHCO), 7.28-7.14 (m, 15H, Ar), 5.38 (d, 1H, J = 6.6 Hz, NHBoc), 4.91 (m, 1H, 1 HOUR),

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4.83 (m, 1H, 1 '' - H), 4.33 (m, 1H, 3'-H), 3.75 (s, 3H, OMe), 3.69 (s, 3H, OMe), 3.34 (m, 2H, 1'-H), 3.31 (m, 2H, 2 '' - H), 3.17 (m, 2H, D 2 '' '- H, 4-CH2), 3.07 (m, 2H, 2' '' - H , 4-CH2), 2.93 (m, 1H, 1'-H), 2.76 (d, 1H, J = 14.7 Hz, 3-H), 2.46 (d, 1H, J = 14.7 Hz, 3-H), 1.86 (m, 1H, 2'-H), 1.60 (m, 1H, 2'-H), 1.43 (s, 9H, CH3 * Bu). 13C NMR (75 MHz, CDCh): 172.7 (COO), 171.8 (COO), 171.6 (2C, CON), 167.4 (CON), 155.8 (CON), 136.3, 136.2, 130.1, 129.4, 129.3, 128.85, 128.8, 128.7, 127.4, 127.3, 127.2 (C, Ar), 80.3 (C, * Bu),

63.3 (C4), 53.6 (2C, C1 '' and C1 '' '), 52.8 (OMe), 52.4 (OMe), 52.0 (C3'), 47.5 (C3), 38.7 (4-CH2), 38.1 (C1 '), 37.9, 37.3 (2C, C2' 'and C2' ''), 31.3 (C2 '), 28.4 (CH3 * Bu). MS (ES) +: 729.53 [M + H] +. Exact mass calculated for C40H48N4O9: 728.34213; Found: 728.34342.

EXAMPLE 47

4S-BenciM- [3’S-ferc-butoxycarbomlammo-3 ’- [W - [(1” S-methoxycarboml-2’-

feml) etM] carbamoM] prop-1’-M] -4- [W - [(1 ’” S-methoxycarboml-2’feml) etM] carbamoil] -2-

oxoazetidine

image109

Sirupe Rdto: 12% (from the compound of Example 3). Semi-preparative HPLC purification (gradient from 50% to 60% of A, in 30 min). HPLC: t R = 15.47 min (gradient from 5% to 100% of A, in 20 min). 1H NMR (400 MHz, CDCh): 5 8.44 (d, 1H, J = 7.6 Hz, NHCO), 7.56 (d, 1H, J = 7.6 Hz, NHCO), 7.32-7.14 (m, 15H, Ar), 5.66 (d, 1H, J = 7.1 Hz, NHBoc), 5.04 (m, 1H, 1 '' - H), 4.76 (m, 1H, 1 '' - H), 4.29 (m, 1H, 3'-H) , 3.73 (s, 3H, OMe), 3.70 (s, 3H, OMe), 3.63 (m, 2H, 1'-H), 3.38 (m, 2H, 4-CH2), 3.35 (m, 2H, 2 ' '-H), 3.16 (dd, 1H, J = 13.9, 5.8 Hz, 2' '' - H), 3.04 (m, 1H, 1'-H), 2.96 (dd, 1H, J = 13.5, 8.2 Hz , 2 '' '- H), 2.60 (dd, 1H, J = 14.8 Hz, 3-H), 2.10 (dd, 1H, J = 14.8 Hz 3-H), 1.92 (m, 2H, 2'-H ), 1.41 (s, 9H, CH3 * Bu). 13C NMR (75 MHz, CDCh): 172.22 (COO), 171.79 (COO), 171.34 (CON), 171.09 (CON), 169.69 (CON), 155.65 (CON), 137.59, 136.41, 135.00, 130.33, 129.44, 129.26 , 128.85, 128.71, 128.68, 127.42, 127.15, 126.98 (C, Ar), 79.86 (C, * Bu), 63.45 (C4), 54.23, 53.41 (2C, C1 '' and C1 ''), 52.46 (OMe ), 52.41 (OMe), 51.87 (C3 '), 46.88 (C3), 40.22 (C1'), 37.87 (4-CH2), 36.61, 36.40 (2C,

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C2 ’’ and C2 ’’), 33.43 (C2 ’), 28.49 (CH3 * Bu). MS (ES) +: 729.53 [M + H] +. Exact mass calculated for C40H48N4O9: 728.34213; Found: 728.34220.

EXAMPLE 48

4R-BenciM- [4’S - [(W-benzyloxycarboml-W-methyl) ammo-4’-methoxycarboml] but-1’-il] -

4- [N - [(1’’S-methoxycarbonyl) ethyl] carbamoyl] -2-oxoazetidine

image110

To a solution of the substituted 4-tert-butoxycarbonyl derivative of Example 10 (0.10 mmol, 55 mg) in CH2Cl2 (1 mL) is added TFA (0.3 mL). After 2 h of reaction at room temperature the solvent is evaporated to dryness, coevaporating several times with CH2Cl2. Subsequently, the corresponding substituted 4-carboxy azetidinone (0.11 mmol, 52 mg) is dissolved in dry THF (1.2 mL) and HL-Ala-OMeHCl (0.22 mmol, 30 mg) is added consecutively , PyBOP (0.22 mmol, 112 mg) and TEA (0.44 mmol, 61 ^ L). After 24 h of stirring at room temperature, the proportion of diastereoisomers formed by HPLC is analyzed. The solvent is then evaporated to dryness. The crude obtained is dissolved in AcOEt and washed with 10% dichloric acid solution, 10% NaHCO3 solution, H2O and saturated NaCl solution. The organic phase is dried over Na2SO4 and evaporated, the resulting residue being purified by semi-preparative HPLC (gradient from 50% to 60% of A, in 30 min) obtaining 20 mg (33%) of a sirupe with the characteristics described at continuation. HPLC: t R = 9.51 min (gradient from 50% to 60% of A, in 15 min). Rotamer ratio M, m = 2: 1 1 H NMR (400 MHz, CDCh): 5 7.35-7.15 (m, 10H, Ph, Z), 6.31 (d, 1H, J = 7.4 Hz, NHCO, M), 6.20 (d, 1H, J = 7.4 Hz, NHCO, m), 5.14 (s, 2H, OCH2), 4.81 (dd, 1H, J = 9.8, 5.0 Hz, 4'-H), 4.56 (m, 1H, 1 '' -H), 3.72 (s, 3H, OMe, M), 3.71 (s, 3H, OMe, M), 3.64 (s, 3H, OMe, m), 3.37 (d, 1H, J = 14.0 Hz, 3-H), 3.25 (m, 2H, 4-CH2), 3.20 (d, 1H, J = 14.0 Hz, 3-H), 2.98 (s, 2H, 1'-H), 2.86 (s, 3H, NCH3), 1.94 (m, 1H, 2'H), 1.76 (m, 3H, 3'-H, 2'-H), 1.32 (d, 3H, J = 7.1 Hz, CH3, M), 1.29 (d , 3H, J = 7.1 Hz, CH3, m). 13C NMR (75 MHz, CDCh): 173.1 (COO), 171.9 (COO, M), 171.2 (COO, m), 170.65 (CON, m), 170.6 (CON, m), 166.5 (C2, M), 166.3 (C2, m), 157.0 (OCON, M), 156.9 (OCON, m), 136.7, 135.2, 135.1, 130.0, 129.9, 129.0, 128.9, 128.7,

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128.6, 128.2, 128.1, 127.9, 127.7, 127.6 (Ar), 67.7 (OCH2, M), 67.6 (OCH2, m), 63.7 (C4), 58.7 (C4 ', m), 58.3 (C4', M), 52.8 (OMe, m) 52.7 (OMe, M), 52.3 (OMe, M), 52.2 (OMe, m), 48.4 (C1``, M), 48.3 (C1 '', m), 46.9 (C3, m ), 46.7 (C3, M), 42.05 (C1 ', m), 42.0 (C1', M), 39.9 (4-CH2, m), 39.8 (4-CH2, M), 31.4 (NMe, m), 30.6 (NMe, M), 26.9 (C3 ', m), 26.5 (C3', M), 25.4 (C2 ', m), 25.2 (C2', M), 18.0 (CH3, Ala). MS (ES) +: 590.40 [M + Na] +.

EXAMPLE 49

4R, S-Benzyl-1 - [(4’S-metMammo-4’-methoxycarboml) but-1’-M] -4- [N - [(1 ”S-methoxycarbonyl) ethyl] carbamoyl] -2-oxoazetidine

image111

A solution of the corresponding substituted 4-benzyloxycarbonyl derivative of Example 48 (0.2 mmol) in MeOH (17 mL) is hydrogenated at room temperature and 15 psi of pressure for 7 h, using Pd-C (10%) as catalyst. Once the catalyst has been filtered off, the solvent is evaporated to dryness, obtaining a sirupe with these characteristics. Rdto: 78%. HPLC: t R = 7.81, 7.95 min (gradient from 2% to 95% of A, in 15 min). Proportion of diastereoisomers

(4S, 4’S, 1’S) :( 4R, 4’S, 1’S) = 27:73. 1H NMR (400 MHz, CDCh): 5 7.31-7.17 (m, 5H, Ph), 6.64 (d, 1H, J = 7.4 Hz, NHCO, M), 6.55 (d, 1H, J = 7.4 Hz, NHCO, m), 4.56 (m, 2H, 4'-H, 1 '' - H,), 3.75 (s, 3H, OMe, M), 3.74 (s, 3H, OMe, m), 3.73 (s, 6H, OMe), 3.40 (d, 1H, J = 14.2 Hz, 4-CH2 M), 3.26 (m, 3H, 1'-H, 4-CH2), 3.23 (d, 1H, J = 14.1 Hz, 4-CH2 M), 3.17 (m, 1H, 1'-H), 3.08 (d, 1H, J = 14.7 Hz, 3-Hm), 2.96 (s, 2H, 3-HM), 2.93 (d, 1H, J = 14.7 Hz, 3-H m), 2.36 (s, 3H, NCH3, m), 2.35 (s, 3H, NCH3, M), 1.85-1.73 (m, 2H, 2'-H), 1.71-1.58 ( m, 2H, 3'-H), 1.41 (d, 3H, J = 7.2 Hz, 2 '' - H, m), 1.33 (d, 3H, J = 7.2 Hz, 2 '' - H, M). 13C NMR (75 MHz, CDCl3): 175.1 (COO, M), 174.9 (COO, m), 173.1 (COO, M), 171.0 (COO, m), 171.0 (CON, m), 170.8 (CON, M) , 166.8 (C2, m), 166.6 (C2, M), 135.0, 129.9, 129.8, 128.7, 128.6, 127.4 (Ar), 63.6 (C4, m), 63.5 (C4, M), 62.6 (C4 ', m ), 62.4 (C4 ', M), 52.7 (OMe), 52.0 (OMe, m), 51.9 (OMe, M), 48.6 (C1' ', m), 48.4 (C1' ', M), 46.9 (C3 , M), 46.8 (C3, m), 42.0 (C1 ', m), 41.9 (C1', M), 39.3 (4-CH2, m),

39.3 (4-CH2, M), 34.7 (NMe, m), 34.6 (NMe, M), 30.7 (C3 ', M), 30.5 (C3', m), 25.0 (C2 ', m), 24.9 (C2 ', M), 18.1 (CH3, m), 18.0 (CH3, M). MS (ES) +: 434.38 [M + H] +.

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EXAMPLE 50

4R-BenciM - [(4’S-benzMoxicarbomlammo-4’-methoxycarboml) but-1’-M] -4- [N - [(1 ”S-

methoxycarbonyl) ethyl] carbamoyl] -2-oxoazetidine

image112

Sirupe Rdto: 27% (from the compound of Example 13). HPLC-MS: t R = 13.02 min (gradient from 2% to 95% of A, in 15 min). Proportion of isomers (4R, 4’S, 1’S) :( 4S, 4’S, 1’S) = 16: 1. 1H NMR (300 MHz, CDCI3, majority isomer): 5 7.10-7.27 (m, 10H, Ar), 6.30 (d, 1H, J = 7.1 Hz, 4'-NH), 5.56 (d, 1H, J = 8.0 Hz, 1 '' - NH),

5.03 (s, 2H, OCH2), 4.48 (quint, 1H, J = 7.1 Hz, 4'-H), 4.30 (m, 1H, 1 ”-H), 3.67 (m, 3H, OCH3), 3.63 (m , 3H, OCH3), 3.22-3.07 (m, 4H, 4-H, 1'-H), 2.91 (s, 2H, 4-CH2), 1.791.60 (m, 4H, 2'-H, 3 ' -H), 1.38 (d, 3H, J = 7.2 Hz, 2 ”-H, m), 1.25 (d, 3H, J = 7.2 Hz, 2” -H, M). MS (ES) +: 554.19 [M + H] +.

EXAMPLE 51

4R, S-BenciM - [(4’S-benzMoxicarbomlammo-4’-methoxycarboml) but-1’-M] -4- [N-

[(1 ”R-methoxycarboml) ethyl] carbamoyl] -2-oxoazetidma

image113

Sirupe Rdto: 44% (from the compound of Example 13). HPLC-MS: t R = 12.98 min (gradient from 2% to 95% of A, in 15 min). Proportion of isomers (4R, 4’S, 1’’R) :( 4S, 4’S, 1’R) = 2: 1. 1H NMR (300 MHz, CDCh): 5 7.27-7.09 (m, 10H, Ar), 6.44 (d, 1H, J = 6.9 Hz, 4'-NH, M), 6.36 (d, 1H, J = 7.0 Hz , 4'-NH, m), 5.50 (d, 1H, J = 8.1 Hz, 1 '' - NH), 5.03 (s, 2H, OCH2), 4.45 (m, 1H, 4'-H), 4.29 ( m, 1H, 1 ”-H), 3.67 (s, 3H, OCH3), 3.64 (s, 3H, OCH3), 3.84-3.34 (m, 6H, 1'-H, 3-H, 4-CH2), 1.90-1.50 (m, 4H, 2'- H, 3'-H), 1.32 (d, 3H, J = 7.2 Hz, 2 ”-H, M), 1.31 (d, 3H, J = 7.2 Hz, 2 "-H, m). MS (ES) +: 554.11 [M + H] +.

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TRPM8 Channel Lock

The efficacy measures of the compounds example 1-example 51 have been carried out using the Chinese hamster ovary cell line (CHO) that stably expresses the TRPM8 protein. The cells were cultured in monolayer in EMEM medium (Earle's minimum essential medium with Earle's salts, Invitrogen) supplemented with 10% FBS, 2 mM L-glutamine, 1% penicillin-streptomycin solution and 0.4 ^ g / mL of antibiotic Geneticina (Sigma) and maintained at 37 ° C in a humidified atmosphere with 5% CO2. To determine the efficacy of the compounds on the TRPM8 channel, Ca2 + fluorimetry tests have been carried out. The cells are seeded in 96-well plates at a density of 25,000 cells / well, after 3 days after planting, the medium is removed and 100 ^ l of the Fluo-4NW fluorescent probe is added at a concentration of 5 ^ M in the presence of 0.02% pluronic acid, after 60 minutes of incubation at 37 ° C in a humidified atmosphere with 5% CO2, fluorescence is measured in a microplate reader (POLARstar Omega) with a configuration of 485 nm for the excitation and 520 nm for the emission, during 20 cycles. During the first 3 cycles the baseline fluorescence is measured and 1 ^ l of the different compounds (examples 1-51) are added to a final concentration of 5 ^ M, after 7 cycles of fluorescence measurement, all wells are added 10 ^ l of menthol to obtain a final concentration of 100 ^ M. In wells that are used as negative controls, 1 ^ l of AMTB (W- (3-Aminopropyl) -2 - [(3-methylphenyl) methoxy] -W- (2-thienylmethyl) benzamide hydrochloride; Tocris Bioscience) is added to obtain a final concentration of 10 ^ M, to block the TRPM8 channels.

Table 1. Results of the activity of the compounds on TRPM8

 Example

 % Lock at 50 ^ M% Lock at 5 ^ M

 EXAMPLE 1

 - 1.01 ± 1.68

 EXAMPLE 2

 - 0.88 ± 0.21

 EXAMPLE 3

 90.26 ± 1.58 75.40 ± 9.93

 EXAMPLE 4

 89.34 ± 2.27 71.44 ± 2.48

 EXAMPLE 5

 91.02 ± 3.34 73.32 ± 2.13

 EXAMPLE 6

 122.25 ± 4.64 74.15 ± 17.04

 EXAMPLE 8

 82.17 ± 29.00 36.86 ± 10.85

 EXAMPLE 9

 - 8.16 ± 13.90

 EXAMPLE 10

 88.94 ± 13.13 19.39 ± 16.99

 EXAMPLE 11

 - 1.99 ± 7.00

 EXAMPLE 12

 - 50.40 ± 17.82

 EXAMPLE 13

 - 24.91 ± 17.80

 EXAMPLE 14

 - 4.37 ± 23.85

 EXAMPLE 15

 - 95.55 ± 2.48

 EXAMPLE 16

 - 87.91 ± 6.97

 EXAMPLE 17

 - 88.66 ± 20.12

 EXAMPLE 19

 25.69 ± 17.41 8.87 ± 7.12

 EXAMPLE 20

 - 103.64 ± 3.13

 EXAMPLE 21

 100.38 ± 3.50 56.89 ± 3.55

 EXAMPLE 22

 91.78 ± 5.09 86.49 ± 5.17

 EXAMPLE 23

 76.89 ± 12.62 50.32 ± 8.34

 EXAMPLE 24

 75.31 ± 13.52 66.23 ± 17.90

 EXAMPLE 25

 - 97.48 ± 5.83

 EXAMPLE 26

 - 93.79 ± 7.97

 EXAMPLE 27

 - 85.81 ± 16.16

 EXAMPLE 28

 - 77.79 ± 13.44

 EXAMPLE 29

 - 68.73 ± 12.78

 EXAMPLE 30

 99.11 ± 4.15 60.08 ± 23.35

 EXAMPLE 31

 - 85.23 ± 7.01

 EXAMPLE 32

 84.16 ± 8.74 37.84 ± 12.33

 EXAMPLE 33

 - 29.88 ± 18.48

 EXAMPLE 34

 - 7.39 ± 8.37

 EXAMPLE 35

 69.98 ± 18.60 41.10 ± 8.87

 EXAMPLE 36

 70.38 ± 15.23 46.49 ± 10.18

 EXAMPLE 37

 103.79 ± 4.93 5.05 ± 3.84

 EXAMPLE 38

 108.06 ± 2.33 4.74 ± 3.45

 EXAMPLE 39

 42.17 ± 11.71 35.31 ± 8.70

 EXAMPLE 40

 39.28 ± 14.67 15.91 ± 22.29

 EXAMPLE 41

 64.56 ± 13.16 43.62 ± 10.44

 EXAMPLE 42

 52.32 ± 7.59 43.05 ± 14.29

 EXAMPLE 43

 43.52 ± 9.82 38.54 ± 12.36

 EXAMPLE 44

 - 3.82 ± 15.23

 EXAMPLE 46

 99.32 ± 3.36 58.91 ± 6.24

 EXAMPLE 47

 - 2.28 ± 4.71

 EXAMPLE 50

 - 11.20 ± 9.30

 EXAMPLE 51

 - 7.80 ± 14.99

5

10

fifteen

twenty

25

30

"Patch-Clamp" essays

For the determination of the IC50 of the selected examples, electrophysiological techniques were used using the "patch-clamp" method with the "wholecell" configuration. The cells were seeded at a density of 250,000 cells / well on round glass covers (18 mm in diameter, thickness 0) in 12-well plates (Costar) and grown in EMEM. After 24 hours, the covers were mounted in a recording chamber (WPI) and were continuously perfused (1 mL / min) with HBSS (140 mM NaCl, 4 mM KCl, 1 mM MgCl2, 1.8 mM CaCl2, D-glucose 5 mM, 10 mM Hepes (all from Sigma), pH 7.4) at «22 ° C. The activity of the TRPM8 channels was evoked by the application of short pulses of 10 s of 100 ^ M menthol, using a gravity-activated perfusion system. The compounds dissolved in the HBSS buffer at different concentrations (from 0.1 to 100 ^ M) were applied in the recording chamber near the cells, keeping the same distance between the cells and the perfusion system in all measurement conditions. Pipette tips for the realization of the measurements were prepared from borosilicate capillaries (World Precision Instruments, Sarasota, FL, USA), using a pipette stretcher (P-97, Sutter Instruments, Novato, CA, USA) to obtain a resistance between 2-4 MQ when filled with internal solution (144 mM KCl, 2mM MgCl2, 5mM EGTA, and 10 mM HEPES, (all from Sigma), pH 7.2).

The currents and voltages were recorded using the total current setting with a frequency of 10 kHz (EPC10 amplifier with Pulse software; HEKA Electronics, Lambrecht, Germany) and analyzed using PulseFit 8.54 software (HEKA, Molecular Devices; WinASCD, G Droogmans, Katholieke Universiteit Leuven, Leuven, Belgium) and Origin 7.5 (OriginLab Corp., Southampton MA, USA).

The different currents, caused by the stimulation of the cells with 100 ^ M of menthol in the presence of the different concentrations of the compounds are subtracted from the current caused by the application with 100 ^ M of menthol in the absence of compounds. The results obtained were adjusted to a sigmoid with the software package of the GraphPad Prism software. The results are shown in FIG. one.

Antitumor activity

5

10

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twenty

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30

35

The compounds of examples 20 and 28 were tested on 60 human tumor cell lines, which include leukemia, melanoma and lung, colon, brain, ovarian, breast, prostate and kidney tumors.

The human tumor cell lines of the cancer detection panel are grown in RPMI 1640 medium containing 5% fetal bovine serum and 2 mM L-glutamine. For a typical selection experiment, the cells are inoculated in 96-well microtiter plates at densities ranging from 5,000 to 40,000 cells / well depending on the doubling time of the cell lines. After cell inoculation, the plates are incubated at 37 ° C, 5% CO2, 95% air and 100% relative humidity, for 24 h before the addition of the experimental drugs.

After 24 h, two plates of each cell line are fixed in situ with trichloroacetic acid (TCA), to have a measure of the cell population for each cell line at the time of drug addition (Tz). The compounds to be tested are solubilized in dimethylsulfoxide at 400 times the maximum desired final test concentration and stored frozen before use. At the time of drug addition, an almuot of the frozen concentrate is thawed and the desired maximum final test concentration is diluted to twice with complete medium containing 50 ^ g / mL gentamicin. Additionally, dilutions are made to provide a total of five concentrations of compound plus control. Alumots of 100 µl of the different dilutions of compound are added to the appropriate wells that already contain 100 µl of medium, giving rise to the required final concentrations.

After the addition of the compounds, the plates are incubated for an additional period of 48 h at 37 ° C, 5% CO2, 95% air, and 100% relative humidity. The test is terminated by the addition of cold TCA. The cells are fixed in situ by the gentle addition of 50 ^ l of 50% (w / v) cold TCA (final concentration, 10% TCA) and incubated for 60 minutes at 4 ° C. The supernatant is discarded, and the plates are washed five times with tap water and air dried. A solution of sulphordamine B (SRB) in 1% acetic acid, (100 ^ l) at 0.4% (w / v) is added to each well, and the plates are incubated for 10 minutes at room temperature. After staining, the unbound dye is washed five times with 1% acetic acid and the plates are air dried. The bound dye is subsequently solubilized with 10 mM of

Trizma base, and the absorbance is read in an automated plate reader at a wavelength of 515 nm. For suspended cells, the methodology is the same except that the assay is terminated by fixing sedimented cells at the bottom of the wells by slowly adding 50 ^ l of 80% TCA 5 (final concentration, 16% TCA). Using the seven absorbance measurements at

zero time, (Tz), as a control of growth, (C), and growth in the presence of the five concentrations of the compounds (Ti)], the percentage of final growth is calculated for each of the compound concentrations and the percentage of tumor growth inhibition.

10

Table 2. Inhibition of tumor cell growth.

Tumor cell line Inhibition of cell growth (%) a, b

Example 20_______________Example 28

 Leukemia

 HL-60 (TB)

 86.94

 MOLT-4

 93.80

 RPMI-8226

 80.44

 MR

 83.37

 Lung (non-small cells)

 A549 / ATCC

 77.21

 EKVX

 81.95

 Colon

 HT29

 74.01

 CNS

 SF-295

 78.61

 Melanoma

 MALME-3M

 75.11

 SK-MEL-5

 106.61

 UAC-257

 80.29

 Ovary

 OVCAR-4

 98.52 71.79

 NCI / ADR-RES

 79.03

 Kidney

 A-498

 113.76 91.51

 ACHN

 77.73

 TK-10

 78.61

 UO-31

 91.26

 Prostate

 PC-3

 85.85 72.17

 Mom

 MCF7

 72.97

 T-47D

 77.72

 MDA-MB-468

 73.94 89.26

aInhibition of the growth of the indicated tumor cells, after adding the compound to a concentration of 10 ^ M. b Only inhibitions greater than

70%

Claims (24)

5 10 fifteen twenty 25 30 1. Compound of formula (I) image 1 R5 (I) where: R1 is selected from (C1-C4) alkyl, substituted or unsubstituted, aralkyl (C1-C4), substituted or unsubstituted, heteroaralkyl (C1-C4), substituted or unsubstituted, aryl, substituted or unsubstituted, and heteroaryl , substituted or unsubstituted; R2 is selected from a group -OR6 and -NR7R8; R3 is selected from hydrogen and (C1-C4) alkyl, substituted or unsubstituted; R4 is selected from hydrogen, (C1-C4) alkyl, substituted or unsubstituted, aralkyl (C1-C4), substituted or unsubstituted, heteroaralkyl (C1-C4), substituted or unsubstituted, aryl, substituted or unsubstituted, and -COR9; R5 is selected from -NR11R10; R8 is selected from hydrogen, (C1-C6) alkyl and aralkyl (C1-C4), heteroaryl, heteroarylalkyl (C1-C4) and -CH (R13) COOR12; R6, R11, R13 and R14 are independently selected from hydrogen, (C1-C6) alkyl, substituted or unsubstituted, aralkyl (C1-C4), substituted or unsubstituted, heteroaralkyl (C1-C4), substituted or unsubstituted, and aryl, substituted or unsubstituted; R7 and R12 are selected from hydrogen and (C1-C6) alkyl, substituted or unsubstituted; R9 is selected from -OR6 and -NR7R8; R10 is selected from hydrogen, (C1-C6) alkyl, substituted or unsubstituted, and (C1-C4) aralkyl, substituted or unsubstituted, COOR14; and n is 1,2, 3 or 4; or any pharmaceutically acceptable salts or stereoisomers thereof. 2. Compound according to claim 1, wherein R1 is (C1-C4) alkyl or aralkyl (C1-C4). 3. Compound according to claim 2, wherein R1 is methyl or benzyl. 5 10 fifteen twenty 25 30 35 4. Compound according to any one of claims 1 to 3, wherein R2 is -OR6 and R6 is hydrogen, (C1-C4) alkyl or benzyl, preferably hydrogen, methyl, tert-butyl or benzyl. 5. Compound according to any one of claims 1 to 3, wherein R2 is -NR7R8; R8 is selected from hydrogen, (C1-C6) alkyl, aralkyl (C1-C4), heteroaryl, heteroaralkyl (C1-C4) and -CH (R13) COOR12 and / or R7 is selected from hydrogen and (C1-C4 alkyl) ). 6. Compound according to the preceding claim, wherein R7 is hydrogen or methyl. 7. Compound according to any of claims 5 or 6, wherein R8 is selected from benzyl, heteroaryl, heteroaralkyl (C1) and -CH (R13) COOR12; R13 is methyl or benzyl and / or R12 is methyl. 8. Compound according to any one of claims 1 to 7, wherein R3 is hydrogen or methyl. 9. Compound according to any of claims 1 to 8, wherein R4 is aralkyl (C1-C4) or -COR9. 10. Compound according to claim 9, wherein R4 is benzyl. 11. Compound according to claim 9, wherein R4 is -COOR14, R14 is hydrogen, (C1-C4) alkyl or benzyl, preferably hydrogen, methyl, tert-butyl or benzyl. 12. Compound according to claim 9, wherein R4 is -CONR7R8; R7 is selected from hydrogen and (C1-C4) alkyl, preferably R7 is H, and / or R8 is selected from hydrogen, (C1-C6) alkyl, aralkyl (C1-C4), heteroaryl, heteroaralkyl (C1-C4) ) and -CH (R13) COOR12 and, preferably R8 is benzyl or -CH (R13) COOCH3, with R13 selected from benzyl, methyl, heteroaralkyl (C1) and heteroaryl. 13. Compound according to any of claims 1 to 12, wherein R5 is -NR11R10, where R11 is independently selected from hydrogen, methyl and benzyl; and / or R10 is selected from hydrogen, methyl, benzyl and COOR14; where R14 is methyl, tert-butyl or benzyl. 5 10 fifteen twenty 25 30 35 14. Compound according to claim 1, selected from the list consisting of: 4R, S-Bendl-4-methoxycarbonyl-1 - [(3'S-tert-butoxycarbonylamino-3'-benzyloxycarbonyl) prop-1'-yl] -2 -oxoazetidine 4R, S-Bendl-4-methoxycarbonyl-1 - [(3'S-tert-butoxycarbonylamino-3'- methyloxycarbonyl) prop-1'-yl] -2-oxoazetidine 4R, S-Bendl-4-bendloxicarbonil-1 - [(3’S-tert-butoxycarbonylamino-3’- benzyloxycarbonyl) prop-1’-il] -2-oxoazetidine 4S-Bendl-4-bendloxicarbonil-1 - [(3’S-tert-butoxycarbonylamino-3’- benzyloxycarbonyl) prop-1’-il] -2-oxoazetidine 4R-Bendl-4-bendloxicarbonil-1 - [(3’S-tert-butoxycarbonylamino-3’- benzyloxycarbonyl) prop-1’-il] -2-oxoazetidine 4R, S-Bendl-4-bendloxicarbonil-1 - [(3’S-tert-butoxycarbonylamino-3’- benzyloxycarbonyl) prop-1’-il] - (3R, Sj-methyl-2-oxoazetidine 4R, S-Methyl-4-methoxycarbonyl-1 - [(3’S-tert-butoxycarbonylamino-3’- benzyloxycarbonyl) prop-1’-il] -2-oxoazetidine 4R, S-Bendl-4-methoxycarbonyl-1 - [(3’S-bendloxicarbonylamino-3’-terc- butoxycarbonyl) prop-1’-il] -2-oxoazetidine 4R, S-Methyl-4-methoxycarbonyl-1 - [(3’S-bendloxicarbonylamino-3’-terc- butoxycarbonyl) prop-1’-il] -2-oxoazetidine 4R, S-Bendl-1- [4’S- (W-Bendloxycarbonyl-W-methyl) amino-4’-methoxycarbonyl] but-1-yl] - 4-tert-butoxycarbonyl-2-oxoazetidine 4R, S-Bendl-1- [4’S - [(W-methyl) amino-4’-methoxycarbonyl] but-1’-il] -4-terc- butoxycarbonyl-2-oxoazetidine 4R, S-Bendl-4-bendloxicarbonil-1 - [(4’S-tert-butoxycarbonylamino-4’- benzyloxycarbonyl) but-1’-yl] -2-oxoazetidine 4R, S-Bendl-N - [(4’S-benzyloxycarbonylamino-4’-methoxycarbonyl) but-1’-il] -4-terc- butoxycarbonyl-2-oxoazetidine 4’S-N - [(4’-Benzyloxycarbonylamino-4’-methoxycarbonyl) but-1’-il] -4R, S-terc- butyloxycarbonyl-4-methyl-2-oxoazetidine 4R, S-Bendl-4-bendloxicarbonil-1 - [(2’S-bendloxicarbonylamino-3’-phenyl) prop-1’-il] - 2-oxoazetidine 4R, S-Bendl-4-bendloxicarbonil-1 - [(2’R-bendloxicarbonylamino-3’-phenyl) prop-1’-il] - 2-oxoazetidine 4S-Bendl-4-bendloxycarbonyl-3S-methyl-1 - [(2’S-bendloxicarbonylamino-3’- phenyl) prop-1’-il] -2-oxoazetidine 5 10 fifteen twenty 25 30 35 4R-Bendl-4-bendloxycarbonyl-3R-methyl-1 - [(2’S-bendloxicarbonylamino-3’- phenyl] prop-1’-il] -2-oxoazetidine 4R-Bendl-4-bendloxycarbonyl-3R-methyl-1 - [(2’R-bendloxicarbonylamino-3’- phenyl) prop-1’-il] -2-oxoazetidine 4S-4-Bendl-4-bendloxicarbonil-3S-methyl-1 - [(2’R-bendloxicarbonylamino-3’- phenyl) prop-1’-il] -2-oxoazetidine 4S-Bendloxicarbonyl-3S, 4S-dimethyl-1 - [(2’R-bendloxicarbonylamino-3’-phenyl) prop-1’- il] -2-oxoazetidine 4R, S-Bendl-4-bendloxicarbonil-1 - [(2’S-dibendlamino-3’-phenyl) prop-1’-il] -2- oxoazetidine 4S-Benzyl -] - (4-benzyloxycarbonyl-3S-methyl-1 - [(2’S-dibenzylamino-3’-phenyl) prop-1’-il] - 2-oxoazetidine 4R-Bendl-4-bendloxicarbonil-3R-methyl-1 - [(2’S-dibendlamino-3’-phenyl) prop-1’-il] -2- oxoazetidine 4R, S-Bendl-4-methoxycarbonyl-1 - [(2’S-dibendlamino-3’-phenyl) prop-1’-il] -2- oxoazetidine 4R, S-Bendl-4-tert-butoxycarbonyl-1 - [(2’S-dibendlamino-3-phenyl) prop-1’-il] -2- oxoazetidine 4S-Bendl-3S-methyl-4-methoxycarbonyl-1 - [(2’S-dibendlamino-3’-phenyl) prop-1’-il] -2- oxoazetidine 4S-Bendl-4-tert-butoxycarbonyl-3S-methyl-1 - [(2’S-dibendlamino-3’-phenyl) prop-1’-il] -2- oxoazetidine 4S-Bendl-4-tert-butoxycarbonyl-3S-methyl-1 - [(2’S-bendlamino-3’-phenyl) prop-1’-il] -2- oxoazetidine 4S-Bendl-4-tert-butoxycarbonyl-3S-methyl-1 - [(2’S-amino-3’-phenyl) prop-1-yl] -2- oxoazetidine 4S-Bendl-4-tert-butoxycarbonyl-3S-methyl-1- [2’S-bendloxycarbonylamino-3’- phenyl) prop-1’-il] -2-oxoazetidine 4S-Bendl-4-tert-butoxycarbonyl-3S-metiM - [(2’S-tert-butoxycarbonylamino-3- phenyl) prop-1’-il] -4-oxoazetidine 4R, S-Bendl-4-carboxy-1- [3’S-tert-butoxycarbonylamino-3’-carboxy) prop-1’-il] -2- oxoazetidine 4R, S-Bendl-1- [4’S-tert-butoxycarbonylamino-4’-carboxy) but-1’-il] -4-carboxy-2- oxoazetidine 4S-Bendl-4 - [(N-bendl) carbamoil] -1 - [(3’S-tert-butoxycarbonylamino-3 ’- [(N- benzyl) carbamoyl] prop-1’-il] -2-oxoazetidine 5 10 fifteen twenty 25 30 4R, S-Bendl-1- [4’S-tert-butoxycarbonylamino-4 ’- [(N-bendl) carbamoil] but-1’-il] -4 - [(N- benzyl) carbamoyl] -2-oxoazetidine 4S-4-Bendl-4 - [(N-bendl) carbamoil] -3S-methyl-1- [3’S-tert-butoxycarbonylamino-3’- [(N-benzyl) carbamoyl] prop-1’-yl] -2-oxoazetidine 4R-4-Bendl-4 - [(N-bendl) carbamoil] -3R-methyl-1- [3’S-tert-butoxycarbonylamino-3’- [(N-benzyl) carbamoyl] prop-1’-yl] -2-oxoazetidine 4S-Bendl-4- [N - [(pyridin-4 ’’ - il) methyl] carbamoil-1 - [(3’S-tert-butoxycarbonylamino-3’- [N - [(pyridin-4 ’- il) methyl] carbamoil] prop-1’-il] -2-oxoazetidine 4R-Bendl-4- [N - [(pyridin-4 ’’ - il) methyl] carbamoil] -1- [3’S-tert-butoxycarbonylamino-3’- [N - [(pyridin-4 ’- il) methyl] carbamoil] prop-1’-il] -2-oxoazetidine 4R, S-Bendl-4 - [(N-pyridin-3-yl) carbamoyl] -1 - [(3’S-tert-butoxycarbonylamino-3 ’- [(N- pyridin-3-yl) carbamoyl] prop-1’-yl] -2-oxoazetidine 4S-Bendl-4 - [(N-pyridin-4-yl) carbamoyl] -1- [3’S-tert-butoxycarbonylamino-3 ’- [(N-pyridin- 4-yl) carbamoyl] prop-1’-il] -2-oxoazetidine 4R-Bendl-4 - [(N- (pyridin-4-yl) carbamoyl] -1- [3’S-tert-butoxycarbonylamino-3 ’- [(N- pyridin-4-yl) carbamoyl] prop-1’-yl] -2-oxoazetidine 4S-Bendl-1- [3’S-tert-butoxycarbonylamino-3 ’- [N - [(1’’S- methoxycarbonyl) ethyl] carbamoyl] prop-1’-il] -4- [N - [(1 ’’ S-methoxycarbonyl) ethyl] carbamoyl] - 2-oxoazetidine 4R-Bendl-1- [3’S-tert-butoxycarbonylamino-3 ’- [N - [(1’’S- methoxycarbonyl) ethyl] carbamoyl] prop-1’-il] -4- [N - [(1 ’’ S-methoxycarbonyl) ethyl] carbamoyl] - 2-oxoazetidine 4R-Bencil-1- [3’S-tert-butoxycarbonylamino-3 ’- [N - [(1’’S-methoxycarbonyl-2’- phenyl) ethyl] carbamoyl] prop-1’-il] -4- [N - [(1 ’’ S-methoxycarbonyl-2’phenyl) ethyl] carbamoyl] -2- oxoazetidine 4S-Bencil-1- [3’S-tert-butoxycarbonylamino-3 ’- [N - [(1’’S-methoxycarbonyl-2’- phenyl) ethyl] carbamoyl] prop-1’-il] -4- [N - [(1 ’’ S-methoxycarbonyl-2’phenyl) ethyl] carbamoyl] -2- oxoazetidine 4R-Benzyl-1- [4’S - [(N-benzyloxycarbonyl-N-methyl) amino-4’-methoxycarbonyl] but-1’-il] -4- [N - [(1’’S-methoxycarbonyl) ethyl] carbamoyl] -2-oxoazetidine 4R, S-Bendl-1 - [(4’S-methylamino-4’-methoxycarbonyl) but-1’-il] -4- [N - [(1’’S- methoxycarbonyl) ethyl] carbamoyl] -2-oxoazetidine 4R-Benzyl-1 - [(4’S-benzyloxycarbonylamino-4’-methoxycarbonyl) but-1’-yl] -4- [N - [(1’’S-methoxycarbonyl) ethyl] carbamoyl] -2-oxoazetidine, and 5 10 fifteen twenty 25 30 4R, S-Benzyl-1 - [(4’S-benzyloxycarbonylamino-4’-methoxycarbonyl) but-1’-il] -4- [N - [(1’’R- methoxycarbonyl) ethyl] carbamoyl] -2-oxoazetidine. 15. Process for obtaining the compound of formula (I) when R2 is -OR6, described according to any one of claims 1 to 4 and 8 to 14, comprising the basic medium cyclization of the compound of formula (II): image2 (II) wherein: R1, R3, R4, R5, R6 and n have been described in any of claims 1 to 4 and 8 to 14. 16. Method of obtaining the compound of formula (I), when R2 is -NR7R8, described according to any of claims 1 to 3 and 5 to 14, comprising the reaction of amines of formula R8R7NH with a carboxylic acid derivative of the compound of formula (I) together with a coupling agetne, wherein the carboxylic acid derivative is a compound of formula (I) when R2 is -OH and R7 and R8 have been described in any of claims 1 to 3 and 5 to 14 . 17. Pharmaceutical composition comprising a compound of formula (I) described according to any one of claims 1 to 14 together with a pharmaceutically acceptable carrier. 18. Use of the compound of formula (I) described according to any of claims 1 to 14 for the preparation of a medicament. 19. Use of the compound of formula (I) described according to any of claims 1 to 14, for the preparation of a medicament for the treatment and / or prevention of a disease associated with alterations of the thermosensory channels, preferably of the TRPM8 channels . 20. Use according to claim 19, wherein said diseases are selected from inflammation, lung disorders, eye diseases, gastrointestinal hypersensitivity, skin sensitivity, neurodegeneration and cancer. 5 21. Use according to claim 20, wherein lung disorders are problems of the respiratory system selected from cough and asthma. 22. Use according to claim 20, wherein the skin sensitivity is selected from 10 pruritus, atopic dermatitis, allergen and psoriasis. 23. Use according to claim 20, wherein the cancer can be selected from melanoma, leukemia, prostate, kidney, breast, pancreas, ovary, lung, colon, central nervous system and osteosarcoma. fifteen 24. Use according to claim 20, wherein the eye diseases are selected from dry eye smdrome and excessive tearing. TRPM8 activity (%) CD CO OR or or 3 No cn L'-l <? Do not image3 Iflu Z9Z8 = oe0l 10 100 1000 [Example 3] (nM) > TRPM8 activity (%) image4 ES 2 600 320 A1