WO2025229179A1

WO2025229179A1 - Use of aminothiolester compounds in combination with hypomethylating agents (hma) for the prevention and treatment of cancer

Info

Publication number: WO2025229179A1
Application number: PCT/EP2025/062070
Authority: WO
Inventors: Mileidys PEREZ; Céline BAIER; Guillaume Martin; Ismail Ceylan
Original assignee: ADVANCED BIODESIGN
Current assignee: ADVANCED BIODESIGN
Priority date: 2024-05-03
Filing date: 2025-05-02
Publication date: 2025-11-06
Anticipated expiration: 2026-11-03

Abstract

The present invention relates to the combination of a compound of formula (I) as described herein with an hypomethylating agent (HMA) for use for the prevention and/ or treatment of cancer, in particular acute myeloid leukemia (AML), and AML-related myeloid diseases. The present invention further relates to a pharmaceutical composition comprising a compound of formula (I) as described herein with an HMA.

Description

Use of aminothiolester compounds in combination with hypomethylating agents (HMA) for the prevention and treatment of cancer

Background

For most patients with cancer, the first line of treatment corresponds to high dose of chemotherapeutic agents, targeted therapies and/or immunotherapeutic approaches. Curative intensive first-in-line approaches are the most important stage in the treatment process, as in many patients it is efficient in debulking the tumour mass and can even led to complete remission or a cure.

Unfortunately, some patients can be refractory to chemotherapeutic, targeted and/or immunomodulatory approaches. Moreso, among cancer patients responding well to the first line treatment, some will eventually relapse or develop therapy-related secondary cancers. All together, these patients with refractory/relapsed (R/R) disease are challenging to treat. In addition, some of patients with cancer are not eligible for the above-mentioned intensive treatments because of either co-morbidities, genetic predispositions, advanced age or systemic toxicities. For all these patients there is still a high unmet medical need.

For example, in the case of haematological malignancies, the incidence of myelodysplastic syndromes (MDS), myeloproliferative neoplasms and acute myeloid leukaemia (AML) is increasing with the aging population. Prognosis and overall survival remain poor in elderly patients. Those who are not eligible for intensive treatment are then treated with hypomethylating agents (Azacitidine (5-azacitidine, AZA) and decitabine (5- aza-2'-deocytidicine, DAC)) in combination with Bcl-2 inhibitors (Venetoclax), or targeted therapies like IDH1 inhibitor (ivosidenib), IDH2 inhibitor (enasidenib) or FLT3 inhibitors (Quizartinib), or Hedgehog pathway inhibitors. These treatments play an important role for AML and MDS patients ineligible to intensive chemotherapy, because of much lower toxicity, but still their efficacy is limited. Similar challenges are encountered by other AML- related diseases, particularly in cases of disease progression, such as chronic myelomonocytic leukemia (CMML), chronic myelogenous leukemia (CML), myeloproliferative neoplasms (MPN), acute leukemias of ambiguous lineage, and secondary AML (sAML), which may arise from a prior history of hematologic malignancy or as a consequence of treatment for non-hematological malignancies (treatment- related AML).

HMA class of drugs are widely used in different anticancer regimes in part due to their relatively good safety profile. However, these drugs are not potent anticancer agents against neither the tumour bulk in general nor against the cancer stem cell populations described as the main responsible for resistance and relapses. Combinations with other therapeutic compounds are often used but these combinations are challenging because of high toxicities brought by the second compound which led to frequent treatment interruptions, discontinuation and/or the only option to resume treatment only with HMA.

The combination of HMA and Bcl-2 protein family inhibitors, such as venetoclax (ABT-199), has emerged as a therapeutic strategy in cancer treatment, particularly for haematological malignancies. Bcl-2 protein family inhibitors target the anti-apoptotic proteins Bcl-2, Mcl-1 , Bcl-xL, BcL-w and/or Bfl-1 , which are often overexpressed in cancer cells, contributing to cell survival and chemotherapy resistance. By inhibiting one or more Bcl-2 protein family, for example with venetoclax (ABT-199), the anti-cancer effects of HMAs are augmented, leading to enhanced efficacy compared to either treatment alone. Both preclinical and clinical studies have demonstrated synergistic effects when combining HMAs with Bcl-2 protein family inhibitors, in particular with venetoclax, resulting in increased apoptosis and improved response rates in patients with challenging diseases such as acute myeloid leukaemia (AML), myelodysplastic syndromes (MDS), and other haematological malignancies. Despite these advancements, in these settings the disease tends to re- emerge and/or progress quickly.

There is thus still a need to find alternative treatments.

In the context of the present invention, the inventors surprisingly found a strong antitumoral, synergistic effect by combining a compound of formula (I) as described herein, with an hypomethylating agent.

The aim of the treatment is to abolish or control the progression of the disease with a synergistic therapeutic effect, which can lead to both, dose and toxic effect reduction, delayed emergency of drug resistance and relapse, thereby prolonging the patient's life and improving comfort and quality of life, and providing an alternative salvage therapy for patients who relapse after Bcl-2 protein family inhibitors plus HMA treatment regimens..

Summary of the invention

The present invention relates to a compound of formula (I): wherein:

R-, and R₂, identical or different, are chosen among a C C₁₀ alkyl group, a phenyl, a benzyl, CHR₅CHR₆OR₄ and (CHR₅)_VOR_4I or RT and R₂ together with the nitrogen atom to which they are linked form an heterocycle, in particular a piperidine or a morpholine; said phenyl and benzyl being optionally substituted by one or more substituents chosen from: linear or branched (Ci-C₇)alkyl, halogen, NO₂ and CONH₂ ;

R₃ is chosen from linear or branched (Ci-C₇)alkyl,

R₄ is chosen from: H, linear or branched (C₂-C₇)alkyl, linear or branched (C₂- C₇)alkenyl, -CONR₇R₈, aryl, heteroaryl, (C₂-C₇)cycloalkyl, linear or branched -(Cr C₇)alkyl-aryl and linear or branched -(C₁-C₇)alkyl-heteroaryl; said aryl, (C₂-C₇)cycloalkyl, and heteroaryl being optionally substituted by one or more substituents chosen from: halogen, linear or branched (C C₇)alkyl optionally substituted by one or more halogen atom, linear or branched (C C₇)alkoxy optionally substituted by one or more halogen atom, -COOH, aryl, -NRR’, -NO₂, or said aryl and heteroaryl being optionally fused to form an heterocycloalkyl;

R₅ and R₆ identical or different are independently chosen from:

• H and linear or branched (CrC₇)alkyl, or

• R₅ and R₆ are linked together to form with the carbon atoms to which they are attached a cycloalkyl, aryl or heteroaryl, or

• R₅ is H and R-i and R₆ are linked together to form with the nitrogen atom linked to R-i an heterocycloalkyl or heteroaryl, or

• R₆ is H and R-, and R₅ are linked together to R-, to form with the nitrogen atom linked to R-, an heterocycloalkyl;

- _v is chosen from 2 to 4;

R₇ is -(C₁-C₃)alkyl;

R₈ is -(C₁-C₃)alkylNRR’;

R and R’ identical or different, are independently chosen from H and linear or branched (C C₇)alkyl, or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers; in combination with an Hypomethylating Agent (HMA), for use for the prevention and/or treatment of cancer in a patient.

The present invention further relates to products comprising a compound of formula (I): wherein:

R-, and R₂, identical or different, are chosen among a Ci-C alkyl group, a phenyl, a benzyl, CHR₅CHR₆OR₄ and (CHR₅)_VOR_4I or RT and R₂ together with the nitrogen atom to which they are linked form an heterocycle, in particular a piperidine or a morpholine; said phenyl and benzyl being optionally substituted by one or more substituents chosen from: linear or branched (C₁-C₇)alkyl, halogen, NO₂ and CONH₂ ;

R₃ is chosen from linear or branched (C₁-C₇)alkyl,

R₅ and R₆ identical or different are independently chosen from:

• H and linear or branched (Ci -C₇)alkyl , or

• R₅ is H and R-, and R₆ are linked together to form with the nitrogen atom linked to R-, an heterocycloalkyl or heteroaryl, or

• R₆ is H and R-, and R₅ are linked together to R-, to form with the nitrogen atom linked to R-, an heterocycloalkyl; v is chosen from 2 to 4; R₇ is -(C₁-C₃)alkyl;

R₈ is -(Ci-C₃)alkylNRR’;

R and R’ identical or different, are independently chosen from H and linear or branched (Ci-C₇)alkyl, or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers; and an Hypomethylating Agent (HMA) as a combined preparation for use spread out over time for use for the prevention and/or treatment of cancer in a patient.

The present invention also relates to pharmaceutical composition comprising a compound of formula (I): wherein:

R-, and R₂, identical or different, are chosen among a C C₁₀ alkyl group, a phenyl, a benzyl, CHR₅CHR₆OR₄ and (CHR₅)_VOR_4I or RT and R₂ together with the nitrogen atom to which they are linked form an heterocycle, in particular a piperidine or a morpholine; said phenyl and benzyl being optionally substituted by one or more substituents chosen from: linear or branched (C C₇)alkyl, halogen, NO₂ and CONH₂ ;

R₃ is chosen from linear or branched (C C₇)alkyl,

R₄ is chosen from: H, linear or branched (C₂-C₇)alkyl, linear or branched (C₂- C₇)alkenyl, -CONR₇R₈, aryl, heteroaryl, (C₂-C₇)cycloalkyl, linear or branched -(Cr C₇)alkyl-aryl and linear or branched -(C₁-C₇)alkyl-heteroaryl; said aryl, (C₂-C₇)cycloalkyl, and heteroaryl being optionally substituted by one or more substituents chosen from: halogen, linear or branched (C C₇)alkyl optionally substituted by one or more halogen atom, linear or branched (Ci-C₇)alkoxy optionally substituted by one or more halogen atom, -COOH, aryl, -NRR’, -NO₂, or said aryl and heteroaryl being optionally fused to form an heterocycloalkyl;

R₅ and R₆ identical or different are independently chosen from:

• H and linear or branched (CrC₇)alkyl, or

• R₅ and R₆ are linked together to form with the carbon atoms to which they are attached a cycloalkyl, aryl or heteroaryl, or • R₅ is H and R-i and R₆ are linked together to form with the nitrogen atom linked to R-i an heterocycloalkyl or heteroaryl, or

- _v is chosen from 2 to 4;

R₇ is -(C₁-C₃)alkyl;

R₈ is -(C₁-C₃)alkylNRR’;

R and R’ identical or different, are independently chosen from H and linear or branched (C₁-C₇)alkyl, or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers; and an Hypomethylating Agent (HMA).

Detailed description of the invention

Compound of formula (I)

The compounds of formula (I) according to the invention are as mentioned above and are described in the patent EP1296946 and in the patent application PCT/EP2020/071640, as well as their process of preparation.

In particular, the compounds of formula (I) are characterized in that R₃ is linear or branched (C C₇)alkyl, preferably methyl, R-, is linear or branched (C C₇)alkyl, preferably methyl, R₂ is a C C₁₀ alkyl group, preferably a methyl or octyl, CHR₅CHR₆OR₄, (CHR₅)_VOR₄ or R-, and R₂ together with the nitrogen atom to which they are linked form an heterocycle, in particular a morpholine, and R₅ and R₆ are H.

For example, R₄ is chosen from H, linear or branched (C₂-C₇)alkyl, linear or branched (C₂-C₇)alkenyl, -CONR₇R₈, (C₂-C₇)cycloalkyl, linear or branched -(C C₇) alkyl-heteroaryl, aryl or benzyl; said (C₂-C₇)cycloalkyl being substituted by one or more substituents chosen from: linear or branched (C C₇)alkyl; said benzyl being optionally substituted by one or more substituents chosen from: linear or branched (C C₇)alkyl optionally substituted by one or more halogen atoms, linear or branched (C C₇)alkoxy optionally substituted by one or more halogen, halogen, or said benzyl optionally being fused to form 1 ,3-benzodioxole.

Alternatively, in particular, R₄ is chosen from H, linear or branched (C₂-C₇)alkyl, linear or branched (C₂-C₇)alkenyl, -(C₁-C₇)alkyl-heteroaryl, aryl, -(C₁-C₇)alkyl-aryl or benzyl; said benzyl being optionally substituted by one or more substituents chosen from: linear or branched (C C₇)alkyl optionally substituted by one or more halogen atoms, linear or branched (C C₇)alkoxy optionally substituted by one or more halogen atoms, halogen or pyridyl, or said benzyl optionally being fused to form 1 ,3-benzodioxole. Alternatively, in particular, R₅ and R₆ are H and R₄ is chosen from H, linear or branched (C₂-C₇)alkyl, linear or branched (C₂-C₇)alkenyl, linear or branched -(Ci-C₇)alkyl-heteroaryl, -(Ci-C₇)alkyl-aryl or benzyl linear or branched; said benzyl being optionally substituted by one or more substituents chosen from: linear or branched (Ci-C₇)alkyl optionally substituted by one or more halogen atoms, linear or branched (C C₇)alkoxy optionally substituted by one or more halogen atoms, halogen.

Alternatively, more particularly, the compounds of formula (I) are characterized in that R₅ and R₆ are H and R₄ is chosen from (C₂-C₇) cycloalkyl, linear or branched -(C₁-C₇)alkyl- heteroaryl, or benzyl; preferably benzyl; said (C₂-C₇) cycloalkyl being substituted by one or more substituents chosen from: linear or branched (C C₇)alkyl, said benzyl being optionally substituted by one or more substituents chosen from: linear or branched (C C₇)alkyl optionally substituted by one or more halogen atom, linear or branched (C C₇)alkoxy optionally substituted by one or more halogen atom, halogen.

As a variant, even more particularly, R-, is methyl and R₄ is chosen from: H, CONR₇R₈ with R₇ being methyl and R₈ being NRR' with R and R' being methyl, ethyl, propene, benzyl, pyridyl, benzyloxybutyl, methyl-cyclohexenyl substituted by one or more methyl, and benzyl substituted by one or more fluorine, chlorine, methoxy or methyl.

As a variant, even more particularly, R-, is a methyl and R₄ is chosen from: H, ethyl, propene, benzyl, pyridyl, benzyloxybutyl and benzyl substituted by one of several fluorine, chlorine, methoxy or methyl.

Even more particularly, said compound of formula (I) is chosen from:

S-methyl 4-[2-ethoxyethyl(methyl)amino]-4-methyl-pent-2-ynethioate;

S-methyl 4-[2-allyloxyethyl(methyl)amino]-4-methyl-pent-2-ynethioate;

S-methyl 4-[2-benzyloxyethyl(methyl)amino]-4-methyl-pent-2-ynethioate;

S-methyl 4-methyl-4-[methyl-[2-(m-tolylmethoxy)ethyl]amino]pent-2- ynethioate;

S-methyl 4-[2-[(3,4-dimethylphenyl)methoxy]ethyl-methyl-amino]-4-methyl- pent-2-ynethioate;

S-methyl 4-[2-[(4-methoxyphenyl)methoxy]ethyl-methyl-amino]-4-methyl- pent-2-ynethioate;

S-methyl 4-[2-[(3,4-dimethoxyphenyl)methoxy]ethyl-methyl-amino]-4-methyl- pent-2-ynethioate;

S-methyl 4-[2-[(3-chlorophenyl)methoxy]ethyl-methyl-amino]-4-methyl-pent-2- ynethioate;

S-methyl 4-[2-[(3-fluorophenyl)methoxy]ethyl-methyl-amino]-4-methyl-pent-2- ynethioate; S-methyl 4-methyl-4-[methyl-[2-(2-pyridylmethoxy)ethyl]amino]pent-2- ynethioate;

S-methyl 4-methyl-4-[methyl-[2-(3-pyridylmethoxy)ethyl]amino]pent-2- ynethioate;

S-methyl 4-methyl-4-[methyl-[2-(4-pyridylmethoxy)ethyl]amino]pent-2- ynethioate;

S-methyl 4-((4-(benzyloxy)butyl)(methyl)amino)-4-methylpent-2-ynethioate;

S-methyl 4-((2-hydroxyethyl)(methyl)amino)-4-methylpent-2-ynethioate;

S-methyl 4-methyl-4-[methyl-[2-(2-naphthylmethoxy)ethyl]amino]pent-2- ynethioate;

S-methyl 4-methyl-4-[methyl-[2-[(2,6,6-trimethylcyclohexen-1- yl)methoxy]ethyl]amino]pent-2-ynethioate; S-methyl 2,5,10,11,11-pentamethyl-6-oxo-7-oxa-2, 5, 10- triazatetradec- 12- yne-14-thioate; S-methyl 4-methyl-4-[methyl(2-phenoxycyclopentyl)amino]pent-2-ynethioate;

(S)-S-methyl 4-(2-((benzyloxy)methyl)pyrrolidin-1-yl)-4-methylpent-2- ynethioate; S-methyl 4-[3(benzyloxy)-1 pyrrolidinyl])-4-methylpent-2-ynethioate;

S-methyl 1- 4-Dimethylamino-4-methyl-pent-2-ynethioate ;

S-methyl 5- 4-Methyl-4-morpholin-4-yl-pent-2-ynethioate;

S-methyl 4-methyl-4- [methyl (octyl) amino] pent-2-ynethioate; or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers.

Still particularly, compound of formula (I) is chosen from:

- S-methyl 1- 4-Dimethylamino-4-methyl-pent-2-ynethioate; and

- S-methyl 4-[2-[(3,4-dimethylphenyl)methoxy]ethyl-methyl-amino]-4-methyl-pent-2- ynethioate, or their pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers.

Unless specified otherwise, the terms used hereabove or hereafter as regards to the compounds of formula (I) have the meaning ascribed to them below:

- _v is chosen from 2 to 4 means that the substituent “CHR₅” is repeated twice CHR5CHR5OR4, three times CHR5CHR5CHR5OR4 or four times CHR5CHR5CHR5CHR5OR4.

“halogen” refers to fluorine, chlorine, brome or iodine atom, in particular fluorine or chlorine atom. "alkyl" represents an aliphatic-hydrocarbon group which may be straight or branched, having 1 to 10, 1 to 7 or 2 to 7 carbon atoms in the chain (Ci-C ) alkyl, (Ci-C₇)alkyl or (C₂-C₇)alkyl, unless specified otherwise. In particular, alkyl groups have 1 to 3 carbon atoms in the chain (Ci-C₃)alkyl. Branched means that one or more alkyl groups such as methyl, ethyl or propyl are attached to a linear alkyl chain. Exemplary alkyl groups include methyl, ethyl, n-propyl, i-propyl, n-butyl, t-butyl, octyl, 2,2-dimethylbutyl, n-pentyl, n-hexyl, n-heptyl, in particular methyl, ethyl or octyl.

"alkenyl" refers to an aliphatic hydrocarbon group containing a carbon-carbon double bond and which may be straight or branched having 2 to 7 carbon atoms in the chain (C₂-C₇)alkenyl, unless specified otherwise. Preferred alkenyl groups have 2 to 3 carbon atoms in the chain (C₂-C₃)alkenyl. Exemplary alkenyl groups include ethenyl, n-propenyl, i-propenyl, n butenyl, i-butenyl, 2,2-dimethylbut-1-enyl, n- pentenyl, in particular propenyl.

“alkoxy” represent an alkyl group as previously defined singular bonded to oxygen. Examples of linear or branched (C C₇)alkoxy include methoxy (CH₃O-) and ethoxy (CH₃CH₂O-) .

"aryl" refers to an aromatic monocyclic or multicyclic hydrocarbon ring system of 6 to 14 carbon atoms, preferably of 6 to 10 carbon atoms. Exemplary aryl groups include phenyl, naphthyl, benzyl, phenanthryl, biphenyl, in particular phenyl.

"heteroaryl" refers to a 5 to 14, preferably 5 to 10 membered aromatic mono-, bi- or multicyclic ring wherein at least one member of the ring is a hetero atom. Hetero atoms can be O or N, in particular N. In particular, each ring comprises from 1 to 3 hetero atoms. Examples include pyrrolyl, pyridyl, piperidinyl, pyrazolyl, pyrimidinyl, pyrazinyl, indolyl, imidazolyl, in particular pyridyl.

“cycloalkyl” refers to a saturated monocyclic or bicyclic non-aromatic hydrocarbon ring of 2 to 7 carbon atoms, preferably 3 to 6 carbon atoms, which can comprise one or more unsaturation. Specific examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl. Preferably, the cycloalkyl group is cyclohexenyl.

“-(C₁-C₇)alkyl-aryl” or “-(C₁-C₇)alkyl-heteroaryl” means that R₄ is linked to the oxygen atom by the carbon of the alkyl group; in particular -(C₁-C₇)alkyl-aryl is a benzyl, -"heterocycle" or "heterocyclloalkyl" refers to a saturated or partially unsaturated non aromatic stable 3 to 14, preferably 5 to 10-membered mono, bi or multicyclic rings which can optionally be bridged and wherein at least one member of the ring is a hetero atom. Typically, heteroatoms include, but are not limited to O or N. In particular, each ring comprises from 1 to 3 hetero atoms. Suitable heterocycles are also disclosed in the Handbook of Chemistry and Physics, 76th Edition, CRC Press, Inc., 1995-1996, pages 225 to 226, the disclosure of which is hereby incorporated by reference. Examples of heterocycloalkyl include, but are not limited to tetrahydropyridyl, tetrahydropyranyl, pyrrolidinyl, piperidyl, morpholinyl, imidazolidinyl, or benzodioxole, in particular 1 ,3 benzodioxole.

The term “substituted” refers to, unless specified otherwise, a substitution with one or more substituents, which may be identical or different, for example chosen from linear or branched (C₁-C₇)alkyl, halogen, NO₂ and CONH₂, linear or branched (C_r C₇)al kyl substituted by one or more halogen atom, linear or branched (C₁-C₇)alkoxy, linear or branched (C₁-C₇)alkoxy substituted by one or more halogen atom, aryl, - COOH, -COOCH₂CH₃, -NRR’, NH₂, NHalkyl and N(alkyl)₂. Examples include in particular methyl, methoxy, chlorine, fluorine, CF₃ and OCF₃.

The compounds of formula (I) as described herein can comprise one or more asymmetric carbon atoms. They can therefore exist in the form of enantiomers or diastereoisomers. These enantiomers and diastereoisomers, as well as their mixtures, including racemic mixtures, form part of the invention.

The compounds of formula (I) as described herein can be provided in the form of a free base or in the form of addition salts with acids, which also form part of the invention.

These salts are advantageously prepared with pharmaceutically acceptable acids, but salts with other acids, useful for example for the purification or for the isolation of the compounds of formula (I) as described herein, also form part of the invention.

As used herein, the expression “pharmaceutically acceptable” refers to those compounds, materials, excipients, compositions or dosage forms which are, within the scope of sound medical judgment, suitable for contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response or other problem complications commensurate with a reasonable benefit/risk ratio.

As used herein, “pharmaceutically acceptable salts” refer to derivatives of the disclosed compounds wherein the parent compound is modified by making acid or base salts thereof. The pharmaceutically acceptable salts include the conventional non-toxic salts or the quaternary ammonium salts of the parent compound formed, for example, from nontoxic inorganic or organic acids. For example, such conventional non-toxic salts include those derived from inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric and the like, including mono, di or tri-salts thereof; and the salts prepared from organic acids such as acetic, propionic, succinic, tartaric, citric, methanesulfonic, benzenesulfonic, glucoronic, glutamic, benzoic, salicylic, toluenesulfonic, oxalic, fumaric, maleic, lactic and the like. Further addition salts include ammonium salts such as tromethamine, meglumine, epolamine, etc., metal salts such as sodium, potassium, calcium, zinc or magnesium.

The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound which contains a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent, or in a mixture of the two. Generally, non-aqueous media like ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred. Lists of suitable salts are found in Remington’s Pharmaceutical Sciences, 20^th ed., Mack Publishing Company, Easton, PA, 2000, the disclosure of which is hereby incorporated by reference.

Process of preparation of compounds of formula (I)

The compounds of formula (I) may be prepared in a number of ways well known to those skilled in the art. The compounds can be synthesized, for example, by application or adaptation of the methods described below, or variations thereon as appreciated by the skilled artisan. The appropriate modifications and substitutions will be readily apparent and well known or readily obtainable from the scientific literature to those skilled in the art.

In particular, the compounds of formula (I) can be prepared according to the processes described in patent EP1296946 and patent application PCT/EP2020/071640.

It will be appreciated that the compounds of formula (I) may contain one or more asymmetrically substituted carbon atoms, and may be isolated in optically active or racemic forms. Thus, all chiral, diastereomeric, racemic forms, isomeric forms of a structure are intended, unless the specific stereochemistry or isomeric form is specifically indicated. It is well-known in the art how to prepare and isolate such optically active forms.

For example, mixtures of stereoisomers may be separated by standard techniques including, but not limited to, resolution of racemic forms, normal, reverse-phase, and chiral chromatography, preferential salt formation, recrystallization, and the like, or by chiral synthesis either from chiral starting materials or by deliberate synthesis of target chiral centers.

Compounds of formula (I) may be prepared by a variety of synthetic routes. The reagents and starting materials are commercially available, or readily synthesized by well- known techniques by one of ordinary skill in the arts. All substituents, unless otherwise indicated, are as previously defined.

In the reactions described hereinafter, it may be necessary to protect reactive functional groups, for example hydroxyl, amino, imino, thio or carboxy groups, where these are desired in the final product, to avoid their unwanted participation in the reactions. Conventional protecting groups may be used in accordance with standard practice, for examples see T.W. Greene and P. G. M. Wuts in Protective Groups in Organic Chemistry, 4th ed.(2007), John Wiley & Sons Inc., 1999; J. F. W. McOmie in Protective Groups in Organic Chemistry, Plenum Press, 1973.

Some reactions may be carried out in the presence of a base. There is no particular restriction on the nature of the base to be used in this reaction, and any base conventionally used in reactions of this type may equally be used here, provided that it has no adverse effect on other parts of the molecule, and unless otherwise indicated. Examples of suitable bases include: sodium hydroxide, potassium carbonate, triethylamine, alkali metal hydrides, such as sodium hydride and potassium hydride; alkyl lithium compounds, such as methyllithium and butyllithium; and alkali metal alkoxides, such as sodium methoxide and sodium ethoxide.

Usually, reactions are carried out in a suitable solvent. A variety of solvents may be used, provided that it has no adverse effect on the reaction or on the reagents involved. Examples of suitable solvents include: hydrocarbons, which may be aromatic, aliphatic or cycloaliphatic hydrocarbons, such as hexane, cyclohexane, benzene, toluene and xylene; amides, such as dimethylformamide; alcohols such as ethanol and methanol and ethers, such as diethyl ether and tetrahydrofuran.

The reactions can take place over a wide range of temperatures. In general, it is found convenient to carry out the reaction at a temperature of from 0°C to 150°C (more preferably from about room temperature to 100°C). The time required for the reaction may also vary widely, depending on many factors, notably the reaction temperature and the nature of the reagents. However, provided that the reaction is effected under the preferred conditions outlined above, a period of from 3 hours to 20 hours will usually suffice.

The compound thus prepared may be recovered from the reaction mixture by conventional means. For example, the compounds may be recovered by distilling off the solvent from the reaction mixture or, if necessary, after distilling off the solvent from the reaction mixture, pouring the residue into water followed by extraction with a water- immiscible organic solvent and distilling off the solvent from the extract. Additionally, the product can, if desired, be further purified by various well-known techniques, such as recrystallization, reprecipitation or the various chromatography techniques, notably column chromatography or preparative thin layer chromatography.

For example, a compound of formula (I) can be obtained by : a) reacting a compound of formula (II) with an organic or inorganic acid b) reacting the compound obtained in step a) with a base ; c) reacting the compound obtained in step b) with CO₂ ; d) reacting the compound obtained in step c) with alkyl chloroformate, a reagent able of forming, with the compound obtained in step c), an acid halide or a reagent able of forming, with the compound obtained in step c), a mixed anhydride ; e) reacting the compound obtained in step d) with an anion precursor compound SMe-; wherein R1 and R2 are as defined herein.

In particular, the base of step b) has a pKa greater than 25, preferably the base used in step b) is selected from lithium or magnesium bases, preferably the base is selected from butyllithium, or hexyllithium.

In particular, the compound of formula (II) is obtained by a step a1) of reaction between 3-chloro-3-methylbut-1-yne with R1 R2NH in an aqueous medium.

In particular, said compound obtained in step a1) is purified by one or more filtrations, for example in filtration or in a succession of 2 to 10 filtrations, preferably in a succession of 2 to 5 filtrations, for example in 4 filtrations.

In one embodiment, 3-chloro-3-methylbut-1-yne is obtained by a reaction step of reacting 2-methylbut-3-yn-2-ol with hydrochloric acid in the presence of a copper catalyst.

In another embodiment, the acid is an inorganic acid chosen from hydrochloric acid, phosphoric acid, nitric acid, sulfuric acid, preferably hydrochloric acid.

In another embodiment, step d) is carried out with:

- an alkyl chloroformate having a (C C₆)alkyl, which may comprise at least one double bond, preferably methyl, ethyl, isoprenyl, tert-butyl or isobutyl chloroformate, preferably isobutyl chloroform ate; or

- a reagent capable of forming with the compound obtained in step c) a mixed anhydride chosen from acid chlorides, for example pivaloyl chloride; or

- a reagent capable of forming, with the compound obtained in step c), an acid halide chosen from SOCI₂, COCI₂, PCI₃, PCI₅, PBr₃ or PPh₃ Br₂.

In one embodiment, the anion precursor compounds SMe- are chosen from the salts of formula XSMe in which X represents an alkali metal or alkaline earth metal, for example Na, methyl mercaptan, or (SMe)₂, preferably NaSMe.

This process is described in detail in the patent application PCT/EP2017/053457, from which the content is incorporated by reference. Alternatively, a compound of formula (I) can be prepared from the corresponding acetylenic amine treated successively by BuLi, COS and Mel. A detailed process of preparation can be found for example in G. Quash et al., European Journal of Medicinal Chemistry 43 (2008) 906-916, from which the content is incorporated by reference, in particular in the part 2 of the Material and Methods section.

The above reactions can be carried out by the skilled person by applying or adapting the methods illustrated in the examples hereinafter.

Further, the process may also comprise the additional step of isolating the compound of formula (I) or (II). This can be done by the skilled person by any of the known conventional means, such as the recovery methods described above.

Generally, the starting products are commercially available mainly from Aldrich or Acros or other typical chemicals supplier or may be obtained by applying or adapting any known methods or those described in the examples.

Lipidic nanocapsule

As mentioned above, in one embodiment, the compound of formula (I) is comprised in a lipidic nanocapsule.

By "lipidic nanocapsule" or NCL, is meant a solvent-free formulation having an oil as solubilizing agent making it possible to encapsulate liposoluble active ingredients.

In the context of the present invention and as mentioned herein, the nanocapsule comprises an oily core in particular liquid/semi-liquid at room temperature and a shell surrounding the oily core in particular rigid at room temperature and whose melting/ transition temperature is high (i.e. in particular between 40°C and 85°C).). The core of the particle is therefore made up of oil, here medium chain triglycerides. The active principle (compound of formula (I)) is solubilized in this phase in the center of the nanocapsule. The surface of the nanocapsule or shell is formed from hydrophilic and lipidic surfactants.

In particular, said nanocapsule comprises: an oily core comprising between 25 and 90% by weight of medium chain triglycerides, preferably between 60 and 80%, relative to the total weight of the nanocapsule, and said compound of formula (I); and a shell surrounding the oily core, comprising between 3 and 25% by weight relative to the total weight of the nanocapsule of at least one lipid surfactant, and at least one hydrophilic surfactant; and in which the ratio by weight relative to the total weight of the nanocapsule between the medium chain triglycerides and said compound of formula (I) is of at least 4. In particular, said nanocapsule has a diameter of between 25 and 115 nm, more particularly between 40 and 80 nm, and for example of 30, 40, 45, 50, 55, 60, 65 or 70 nm.

This size makes it possible in particular to ensure optimum efficiency of the nanocapsule in vivo.

In particular, said nanocapsule has a PDI or polydispersity index below 0.2, in particular below 0.1. This index, which is also called dispersity, has the common sense known to those skilled in the art, i.e. the measure of the heterogeneity of the sizes of molecules or particles in a mixture. This allows in particular that the size distribution of the nanocapsules is monodisperse.

As mentioned above, the oily core comprises between 25 and 90% by weight of medium chain triglycerides, preferably between 60 and 80%, relative to the total weight of the nanocapsule, and for example, 40, 50, 55, 60 or 70 % by weight of medium chain triglycerides.

By "medium chain triglycerides" is meant the common sense known to those skilled in the art, i.e. triglycerides in which the three hydroxyl groups of glycerol are esterified with medium chain fatty acids.

The term “medium chain” is understood to mean in particular chains of 6 to 12 carbon atoms.

They are notably present in coconut oil, palm oil and butter.

In particular, the medium chain triglycerides are chosen from mixtures of triglycerides of saturated fatty acids, in particular of caprylic acid (octanoic) and of capric acid (decanoic).

The fatty acids can be obtained from the oil extracted from the hard, dried fraction of the albumen of Cocos nucifera L. or from the oil extracted from the dried albumen of Elaeis guineensis Jacq. meeting the prerequisites of the European Pharmacopoeia such as, more particularly Kollisolv® MCT 70, MIGLYOL® 810 N, MIGLYOL® 812 N Excipient and Labrafac®, preferably MIGLYOL® 812 N

In particular, the oily core comprises between 25 and 90% by weight of the MIGLYOL® 812 N preferably between 60 and 80%, relative to the total weight of the nanocapsule; for example 40, 50, 55, 60 or 70%.

As mentioned above, the shell surrounding the oily core comprises between 3 and 25% by weight based on the total weight of the nanocapsule of at least one lipidic surfactant. In particular, the shell comprises between 3 and 20% by weight relative to the total weight of the nanocapsule of at least one lipidic surfactant, more particularly between 5 and 20%, for example 5, 6, 8, 10, 12 or 20%. This proportion of lipidic surfactant makes it possible in particular to improve the stability of the nanocapsule during its storage at low temperature. By low temperature is meant here a temperature below 0 ° C.

By "lipidic surfactant" is meant the common sense known to those skilled in the art, i.e. an emulsifying agent having an increased tropism for lipid solutions and making it possible to stabilize the nanocapsule.

In particular, said at least one lipidic surfactant is chosen from lecithins obtained from soybean oil having a percentage of phosphatidylcholine greater than 90% corresponding to the prerequisite of the European Pharmacopoeia, in particular, Lipoid®, preferably Lipoid® S100.

More particularly, the shell comprises between 3 and 20% by weight relative to the total weight of the nanocapsule of Lipoid® S100, more particularly between 5 and 20%, for example 5, 6, 8, 10, 12 or 20%.

As mentioned above, the shell surrounding the oily core also includes at least one hydrophilic surfactant.

By "hydrophilic surfactant" is meant the common sense known to those skilled in the art, i.e. an amphiphilic molecule and/or emulsifying agent allowing and stabilizing the nanoparticles and making the latter soluble in water.

In particular, said at least one hydrophilic surfactant is chosen from nonionic solubilizing and/or emulsifying agents meeting the prerequisite described in the European Pharmacopoeia, in particular Macrogol 15 Hydroxystearate, more particularly Kolliphor®, preferably Kolliphor® HS15.

In particular, the shell comprises between 15 and 60% by weight relative to the total weight of the nanocapsule of at least one hydrophilic surfactant, more particularly between 20 and 50%, for example 15; 20; 30; 35; 40 or 50%.

More particularly, the shell comprises between 15 and 60% by weight relative to the total weight of the nanocapsule of Kolliphor® HS15, more particularly between 20 and 50%, for example 15; 20; 30; 35; 40 or 50%.

In particular, in the context of the present invention, said compound of formula (I) can be comprised in the nanocapsule at a concentration of between 5 and 15 mg/mL, more particularly between 8 and 13 mg/mL, for example of 9, 10 or 11 mg/mL.

This concentration makes it possible in particular to prevent the crystallization of the compound of formula (I) over time while limiting its hydrolysis.

As mentioned above, the nanocapsule according to the invention comprises a ratio by weight relative to the total weight of the nanocapsule between the medium chain triglycerides and said compound of formula (I) of at least 4, in particular of at least 5. By “at least 4”, is meant in particular between 4 and 100, more particularly between 5 and 50, for example 4, 5, 6, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65,70, 75, 80, 85, 90, 95 or 100.

By “at least 5”, is meant in particular between 5 and 100, more particularly between 10 and 50, for example 5, 6, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65,70, 75, 80, 85, 90, 95 or 100.

This ratio makes it possible in particular to avoid crystallization of the compound of formula (I).

Thus, a nanocapsule according to the invention can for example be characterized in that it comprises:

- an oily core comprising between 25 and 90% by weight of Labrafac ®, preferably between 60 and 80%, relative to the total weight of the nanocapsule; and

- a shell surrounding the oily core comprising between 3 and 25% by weight relative to the total weight of the nanocapsule of Lipoid ® S100, and between 15 and 60% by weight relative to the total weight of the nanocapsule of Kolliphor® HS15, more particularly between 20 and 50%, for example 15; 20; 30; 35; 40 or 50%;

• in which the oily core comprises one compound of formula (I) as mentioned above;

• in which the ratio by weight relative to the total weight of the nanocapsule between the Labrafac ® and said compound of formula (I) is of at least 4, preferably at least 5.

In particular said compound of formula (I) is S-methyl 4-[2-ethoxyethyl(methyl)amino]- 4-methyl-pent-2-ynethioate; or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers.

In particular said compound of formula (I) is S-methyl 4-[2- allyloxyethyl(methyl)amino]-4-methyl-pent-2-ynethioate; or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers.

In particular said compound of formula (I) is S-methyl 4-[2- benzyloxyethyl(methyl)amino]-4-methyl-pent-2-ynethioate; or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers.

In particular said compound of formula (I) is S-methyl 4-methyl-4-[methyl-[2-(m- tolylmethoxy)ethyl]amino]pent-2-ynethioate; or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers.

In particular said compound of formula (I) is S-methyl 4-[2-[(3,4- dimethylphenyl)methoxy]ethyl-methyl-amino]-4-methyl-pent-2-ynethioate; or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers. In particular said compound of formula (I) is S-methyl 4-[2-[(4- methoxyphenyl)methoxy]ethyl-methyl-amino]-4-methyl-pent-2-ynethioate; or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers.

In particular said compound of formula (I) is S-methyl 4-[2-[(3,4- dimethoxyphenyl)methoxy]ethyl-methyl-amino]-4-methyl-pent-2-ynethioate; or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers.

In particular said compound of formula (I) is S-methyl 4-[2-[(3- chlorophenyl)methoxy]ethyl-methyl-amino]-4-methyl-pent-2-ynethioate; or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers.

In particular said compound of formula (I) is S-methyl 4-[2-[(3- fluorophenyl)methoxy]ethyl-methyl-amino]-4-methyl-pent-2-ynethioate; or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers.

In particular said at compound of formula (I) is S-methyl 4-methyl-4-[methyl-[2-(2- pyridylmethoxy)ethyl]amino]pent-2-ynethioate; or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers.

In particular said compound of formula (I) is S-methyl 4-methyl-4-[methyl-[2-(3- pyridylmethoxy)ethyl]amino]pent-2-ynethioate; or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers.

In particular said compound of formula (I) is S-methyl 4-methyl-4-[methyl-[2-(4- pyridylmethoxy)ethyl]amino]pent-2-ynethioate; or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers.

In particular said compound of formula (I) is S-methyl 4-((4- (benzyloxy)butyl)(methyl)amino)-4-methylpent-2-ynethioate; or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers.

In particular said compound of formula (I) is S-methyl 4-((2- hydroxyethyl)(methyl)amino)-4-methylpent-2-ynethioate; or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers.

In particular said compound of formula (I) is S-methyl 4-methyl-4-[methyl-[2-(2- naphthylmethoxy)ethyl]amino]pent-2-ynethioate; or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers. In particular said compound of formula (I) is S-methyl 4-methyl-4-[methyl-[2-[(2,6,6- trimethylcyclohexen-1- yl)methoxy]ethyl]amino]pent-2-ynethioate; or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers.

In particular said compound of formula (I) is S-methyl 2,5,10,11 ,11-pentamethyl-6- oxo-7-oxa-2,5,10-triazatetradec-12-yne-14-thioate; or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers.

In particular said compound of formula (I) is S-methyl 4-methyl-4-[methyl(2- phenoxycyclopentyl)amino]pent-2-ynethioate; or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers.

In particular said compound of formula (I) is (S)-S-methyl 4-(2- ((benzyloxy)methyl)pyrrolidin-1-yl)-4-methylpent-2-ynethioate; or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers.

In particular said compound of formula (I) is S-methyl 4-[3(benzyloxy)-1pyrrolidinyl])- 4-methylpent-2-ynethioate; or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers.

In particular said compound of formula (I) is S-methyl 1- 4-Dimethylamino-4-methyl- pent-2-ynethioate; or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers.

In particular said compound of formula (I) is S-methyl 5- 4-Methyl-4-morpholin-4-yl- pent-2-ynethioate; or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers.

In particular said compound of formula (I) is S-methyl 4-methyl-4- [methyl (octyl) amino] pent-2-ynethioate; or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers.

Preparation of the lipidic nanocapsule

The lipidic nanocapsules can be prepared as mentioned in the experimental part.

For example, the nanocapsules are produced as follows:

Weigh the medium chain triglycerides, add the compound of formula (I) and leave under steering for mixing;

Weigh said at least one hydrophilic surfactant and said at least one lipidic surfactant, add NaCI and water;

Transfer the aqueous phase into the oily phase under steering;

Transfer into the oil bath and heat up the solution under strong steering; Once a certain temperature is reached, transfer into the ice to cool down ;

T ransfer back to the oil bath to heat ;

Repeat cooling and heating steps additional times;

- At the end of the last heating step, transfer into the ice bath and cool down ;

When the formulation reaches a certain temperature, add cold water ;

Remove the ice bath and leave under agitation ;

Transfer the formulation into an appropriate container.

In one embodiment, the nanocapsule can be formulated in a formulation comprising water and a salt, such as NaCI.

Thus this formulation comprises the nanoparticle, water, preferably between 50 and 70%, and for example 60%, by weight relative to the total weight of the formulation, and a salt, preferably between 1 and 5%, for example 2 or 3%, by weight, relative to the total weight of the composition.

HMA

HMA refers to a specific class of nucleoside analogs capable of incorporating into DNA and/or RNA, where they exert direct inhibition of DNA methyltransferases (DNMTs). This leads to the passive loss of DNA methylation during replication, resulting in transient DNA hypomethylation both in vitro and in vivo.

They refer to a group of agents with the capacity, both in vitro and in vivo, to induce transient DNA hypomethylation. DNA methylation refers to the addition of a methyl group to a CpG site. These sites cluster together in areas known as CpG islands and are frequently localized in the proximity of key gene regulatory regions such as gene promoters. DNA methylation, both aberrant and physiologic, of these areas can result in gene silencing and in the equivalent of the physical inactivation, due to either mutations or deletions, of tumor suppressor genes.

Examples of HMA according to the invention include, but are not limited to, Azacitidine, Decitabine, Guadecitabine, CC-486, SGI-110, CP-4200, NTX-301 , ASTX727. They also include other HMA formulations, if the combination according to the invention further includes a cytidine deaminase inhibitor.

Azacitidine or decitabine are particularly contemplated when the cancer to treat is AML, myelodysplastic syndromes (MDS), chronic myelomonocytic leukemia (CMML), myeloproliferative neoplasms (MPNs) in transformation or with a dysplasia such as Post- MPN AML, myelofibrosis with excess of blasts (MF-EB), pre-leukemia phases of primary myelofibrosis, juvenile myelomonocytic leukjemia (JMML), chronic myelogenus leukemia (CML) with blast crisis (myeloid phenotype), acute lymphoblastic leukemia (ALL), particularly in T-ALL or relapse/refractory disease, blastic plasmacytoid dendritic cell neoplasm (BPDCN), and T-cell lymphomas, especially AML.

Guadecitabine is particularly contemplated when the cancer to treat is leukemia and certain solid tumors, including ovarian cancer, pancreatic cancer, colorectal cancer, and non-small cell lung cancer (NSCLC).

Examples of cytidine deaminase inhibitors according to the invention includes Tetrahydrouridine (THU) and 3,4-Dihydro-5-[4-(1-piperidinyl)butoxy]-4-oxo-1(2H)- pyrimidine (CP-4055). Those compounds are commercially available.

In particular, cisplatin and ATO (arsenic trioxide) are not HMA according to the invention.

Pharmaceutical composition

As previously mentioned, the present invention relates to a pharmaceutical composition comprising a compound of formula (I) as described herein or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers as described herein; and an Hypomethylating Agent (HMA) as described herein, optionally by admixture with one or more pharmaceutically acceptable excipients.

In one embodiment, said pharmaceutical composition further comprises a cytidine deaminase inhibitor.

Such composition may be prepared for use in oral administration, particularly in the form of tablets or capsules, in particular orodispersible (lyoc) tablets; or parenteral administration e.g. subcutaneous, intramuscular, intravenous, intradermal, particularly in the form of liquid solutions, suspensions or emulsions.

It may be prepared by any of the methods well known in the pharmaceutical art, for example, as described in Remington: The Science and Practice of Pharmacy, 20^th ed.; Gennaro, A. R., Ed.; Lippincott Williams & Wilkins: Philadelphia, PA, 2000. Pharmaceutically compatible binding agents and/or adjuvant materials can be included as part of the composition. Oral compositions will generally include an inert diluent carrier or an edible carrier. They can be administered in unit dose forms, wherein the term “unit dose” means a single dose which is capable of being administered to a patient, and which can be readily handled and packaged, remaining as a physically and chemically stable unit dose comprising either the active compound itself, or as a pharmaceutically acceptable composition.

The tablets, pills, powders, capsules, troches and the like can contain one or more of any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, or gum tragacanth; a diluent such as starch or lactose; a disintegrant such as starch and cellulose derivatives; a lubricant such as magnesium stearate; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, or methyl salicylate. Capsules can be in the form of a hard capsule or soft capsule, which are generally made from gelatin blends optionally blended with plasticizers, as well as a starch capsule. In addition, dosage unit forms can contain various other materials that modify the physical form of the dosage unit, for example, coatings of sugar, shellac, or enteric agents. Other oral dosage forms syrup or elixir may contain sweetening agents, preservatives, dyes, colorings, and flavorings. In addition, the active compounds may be incorporated into fast dissolve, modified-release or sustained-release preparations and formulations, and wherein such sustained-release formulations are preferably bi-modal.

Liquid preparations for administration include sterile aqueous or non-aqueous solutions, suspensions, and emulsions. The liquid compositions may also include binders, buffers, preservatives, chelating agents, sweetening, flavoring and coloring agents, and the like. Non-aqueous solvents include alcohols, propylene glycol, polyethylene glycol, acrylate copolymers, vegetable oils such as olive oil, and organic esters such as ethyl oleate. Aqueous carriers include mixtures of alcohols and water, hydrogels, buffered media, and saline. In particular, biocompatible, biodegradable lactide polymer, lactide/glycolide copolymer, or polyoxyethylene-polyoxypropylene copolymers may be useful excipients to control the release of the active compounds. Intravenous vehicles can include fluid and nutrient replenishers, electrolyte replenishers, such as those based on Ringer's dextrose, and the like.

Use

As already mentioned, the present invention relates to a compound of formula (I) as described herein (I) or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers as described herein; in combination with an Hypomethylating Agent (HMA) as described herein, for use for the prevention and/or treatment of cancer in a patient.

The present invention further relates to products comprising a compound of formula (I) as described herein (I) or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers as described herein; in combination with an Hypomethylating Agent (HMA) as described herein, as a combined preparation for use spread out over time for use for the prevention and/or treatment of cancer in a patient. The present invention further relates to a method to prevent and/or treat cancer in a patient in need thereof, comprising the administration of a compound of formula (I) as described herein or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers as described herein; in combination with an Hypomethylating Agent (HMA) as described herein.

The term “patient" can be used as well as “subject” and refers to a warm-blooded animal such as a mammal, in particular a human, male or female, unless otherwise specified, which is afflicted with, or has the potential to be afflicted with one or more diseases and conditions described herein.

In one embodiment, said patient has a cancer and is not eligible for intensive chemotherapeutic treatments because of either co-morbidities, genetic predispositions, advanced age, or systemic toxicities.

In particular, by “intensive chemotherapeutic treatments” is meant any regime including a chemotherapeutic agent: e.g alkylating agents such as cyclophosphamide, cisplatin, carboplatin, busulfan, among others; antimetabolites such as fluorouracil, capecitabine, methotrexate, gemcitabine, among others; a nth racy clines such as doxorubicin and daunirubicin, epirubicin, idarubicin; other antitumor antibiotics such as bleomycin, mitomycin C; topoisomerase inhibitors such as etoposide, irinotecan, topotecan; mitotic inhibitors such as paclitaxel, docetaxel, vincristine, vinblastine; hormonal agents and targeted-therapies.

The terms "treat", “treating”, “treated” or "treatment", as used in the context of the invention, refer to therapeutic treatment wherein the object is to eliminate or lessen symptoms. Beneficial or desired clinical results include, but are not limited to, elimination of symptoms, alleviation of symptoms, diminishment of extent of condition, stabilized (i.e., not worsening) state of condition, delay or slowing of progression of the condition.

The terms “prevent”, “prevention”, “preventing” or “prevented”, as used in the context of the present invention, refer to the prevention of the onset, recurrence or spread of a disease or disorder, or of one or more symptoms thereof. In certain embodiments, the terms refer to the treatment with or administration of a compound provided herein prior to the onset of symptoms, particularly to patients at risk of disease or disorders provided herein. The terms encompass the inhibition or reduction of a symptom of the particular disease. Subjects with familial history of a disease in particular are candidates for preventive regimens in certain embodiments. In addition, subjects who have a history of recurring symptoms are also potential candidates for the prevention. In this regard, the term “prevention” may be interchangeably used with the term “prophylactic treatment”. In the context of the present invention, the identification of the subjects who are in need of treatment of herein-described diseases and conditions is conducted as above mentioned and is well within the ability and knowledge of the man skilled in the art. A clinician skilled in the art can readily identify, by the above-mentioned technics, those subjects who are in need of such treatment.

As used herein and unless otherwise defined, "cancer" refers to the growth, division or proliferation of abnormal cells in the body.

In particular, said cancer is a relapsed or refractory cancer.

By “relapsed” is meant a cancer that has returned or progressed after a period of remission or stable disease following an initial response to treatment. A relapse cancer may occur at the same site as the original tumor (local recurrence) or may spread to other parts of the body (metastatic recurrence).

By “refractory” is meant a cancer that does not respond to treatment or stops responding after an initial response. In other words, it is cancer that continues to grow or progress despite undergoing one or more courses of standard therapy.

In one embodiment, the patient has been previously treated with a chemotherapybased regimen, in particular an anthracycline based regimen such as daunorubicin, idarubicin, doxorubicin or epirubicin.

As such, the treatment according to the invention (i.e the administration of a compound of formula (I) or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers and HMA as described herein) can be a first-line treatment, a second-line treatment, a third-line treatment or a maintenance treatment.

By "maintenance treatment" is meant, in the context of cancer therapy, an ongoing treatment administered after a patient has completed initial therapy, such as surgery, chemotherapy, or radiation therapy. The goal of maintenance treatment is to prolong the duration of remission or to prevent the recurrence of cancer.

Cancers according to the invention are in particular chosen from hematological and solid cancers. More particularly, said cancer is chosen from Acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), chronic myelomonocytic leukemia (CMML), myeloproliferative neoplasms (MPNs) in transformation or with a dysplasia such as Post- MPN AML, myelofibrosis with excess of blasts (MF-EB), pre-leukemia phases of primary myelofibrosis, juvenile myelomonocytic leukemia (JMML), chronic myelogenus leukemia (CML) with blast crisis (myeloid phenotype), acute lymphoblastic leukemia (ALL), particularly in T-ALL or relapse/refractory disease, blastic plasmacytoid dendritic cell neoplasm (BPDCN), T-cell lymphomas, Lymphoma non-Hodgkin, Large B-Cell Diffuse Lymphoma, Lung cancer, Colorectal cancer, including microsatellite stable colorectal cancer, Breast Cancer, including patients with advanced or metastatic HER2-negative breast cancer, and patients with triple-negative breast cancer, Liver and hepatic Cancer, Prostate cancer, Epithelial Ovarian Cancer, Pancreatic and oesophageal/gastric adenocarcinoma (EGC), Skin Neoplasm, Kidney/Renal Cancer, Urothelial or Bladder Carcinoma Anorexia Nervosa, Testis Cancer, and Glioblastoma.

In the context of the invention, the prevention and/or treatment of AML, MDS, , chronic myelomonocytic leukemia (CMML), myeloproliferative neoplasms (MPNs) in transformation or with a dysplasia such as Post-MPN AML, myelofibrosis with excess of blasts (MF-EB), pre-leukemia phases of primary myelofibrosis, juvenile myelomonocytic leukemia (JMML), chronic myelogenus leukemia (CML) with blast crisis (myeloid phenotype), acute lymphoblastic leukemia (ALL), particularly in T-ALL or relapse/refractory disease, blastic plasmacytoid dendritic cell neoplasm (BPDCN), and T-cell lymphomas is particularly contemplated, especially AML, MDS and MPN.

Still particularly, in the context of the prevention and/or treatment of AML, chronic myelomonocytic leukemia (CMML), myeloproliferative neoplasms (MPNs) in transformation or with a dysplasia such as Post-MPN AML, myelofibrosis with excess of blasts (MF-EB), pre-leukemia phases of primary myelofibrosis, juvenile myelomonocytic leukemia (JMML), chronic myelogenus leukemia (CML) with blast crisis (myeloid phenotype), acute lymphoblastic leukemia (ALL), particularly in T-ALL or relapse/refractory disease, blastic plasmacytoid dendritic cell neoplasm (BPDCN), and T-cell lymphomas, and especially of AML, MDS and MPN, compound of formula (I) as described herein is chosen from S-methyl 1- 4-Dimethylamino-4-methyl-pent-2-ynethioate and S-methyl 4-[2-[(3,4- dimethylphenyl)methoxy]ethyl-methyl-amino]-4-methyl-pent-2-ynethioate, or their pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers, and is more particularly S-methyl 1- 4-Dimethylamino-4-methyl- pent-2-ynethioate or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers.

In the context of the invention, the prevention and/or treatment of AML is more particularly contemplated.

Still particularly, in the context of the prevention and/or treatment of AML, compound of formula (I) as described herein is S-methyl 1- 4-Dimethylamino-4-methyl-pent-2- ynethioate or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers.

In one embodiment, the patient is also treated with a Bcl2 protein family inhibitor, in particular venetoclax (ABT-199). In such case, the treatment according to the invention (i.e the administration of a compound of formula (I) or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers and HMA as described herein) can be a second or third-line treatment.

By “Bcl2 protein family inhibitor”, is meant in particular Bcl2 inhibitors and MCL1 inhibitors. Venetoclax (ABT-199), Navitoclax (ABT-263), Obatoclax (GX15-070), Oblimersen socium (G3139), AMG 176, AMG397, AZD5991 , S64315 (MIK665), VU661013, Gossypol (AT-101), Paclitaxel and Docetaxel can for example be cited.

Ventoclax is particularly contemplated if the cancer to treat is AML.

In one embodiment, the patent is also administered with a cytidine deaminase inhibitor, for example [to complete with the same examples than those

A therapeutically effective amount can be readily determined by the attending diagnostician, as one skilled in the art, by the use of conventional techniques and by observing results obtained under analogous circumstances. In determining the therapeutically effective amount, a number of factors are considered by the attending diagnostician, including, but not limited to: the species of subject; its size, age, and general health; the specific disease involved; the degree of involvement or the severity of the disease; the response of the individual subject; the particular compound administered; the mode of administration; the bioavailability characteristic of the preparation administered; the dose regimen selected; the use of concomitant medication; and other relevant circumstances.

As used herein, an «effective amount" refers to an amount which is effective in reducing, eliminating, treating or controlling the symptoms of the herein-described diseases and conditions. The term "controlling" is intended to refer to all processes wherein there may be a slowing, interrupting, arresting, or stopping of the progression of the diseases and conditions described herein, but does not necessarily indicate a total elimination of all disease and condition symptoms, and is intended to include prophylactic treatment and chronic use.

The amount of the compounds according to the invention (i.e. compound of formula (I) or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers and HMA as described herein), which is required to achieve the desired biological effect, will vary depending upon a number of factors, including the dosage of the drug to be administered, the chemical characteristics (e.g. hydrophobicity) of the compounds employed, the potency of the compounds, the type of disease, the diseased state of the patient, and the route of administration.

Compound of formula (I) or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers and HMA provided herein can be formulated into pharmaceutical compositions, optionally by admixture with one or more pharmaceutically acceptable excipients.

Such compositions may be prepared for use in oral administration, particularly in the form of tablets or capsules, in particular orodispersible (lyoc) tablets; or parenteral administration, e.g. subcutaneous, intramuscular, intravenous, intradermal, particularly in the form of liquid solutions, suspensions or emulsions.

Liquid preparations for administration include sterile aqueous or non-aqueous solutions, suspensions, and emulsions. The liquid compositions may also include binders, buffers, preservatives, chelating agents, sweetening, flavoring and coloring agents, and the like. Non-aqueous solvents include alcohols, propylene glycol, polyethylene glycol, acrylate copolymers. Aqueous carriers include mixtures of alcohols and water, hydrogels, buffered media, and saline. In particular, biocompatible, biodegradable lactide polymer, lactide/glycolide copolymer, or polyoxyethylene-polyoxypropylene copolymers may be useful excipients to control the release of the active compounds. Intravenous vehicles can include fluid and nutrient replenishers, electrolyte replenishers, such as those based on Ringer's dextrose, and the like.

In particular, said compound of formula (I) or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers and HMA as described herein are administered separately, sequentially or simultaneously, preferably sequentially.

For example, if said compound of formula (I), encapsulated or not, and said HMA are administered sequentially, the HMA can be administered prior to administration of the compound of formula (I) or pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers. In particular, the HMA is administered orally and the compound of formula (I) or pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers by parenteral or subcutaneous administration.

In the context of the present invention, it should be understood that "a compound for use for the prevention or treatment of' is equivalent to "the use of a compound for the prevention or treatment of" and to "the use of a compound for the manufacture of a medicament for the prevention or treatment of ”.

The invention will be further illustrated by the following examples.

Note: in the following figures and examples, DI MATE corresponds to S-methyl 1- 4- Dimethylamino-4-methyl-pent-2-ynethioate.

Figures

Figure 1 : Characteristic synergistic interaction map obtained for the combination of DI MATE and the hypomethylating agents (HMA) Azacitidine and decitabine, in different AML cell lines as indicated in the figure.

Figure 2: Dendrogram of Drug-drug interaction between 10 different standard of care compounds and DIMATE in 10 AML cell lines with different genetic background.

Figure 3: Evaluation of the efficacy of DIMATE alone and in combination with hypomethylating agents (HMA) in hematopoietic colony formation assays using leukemic cells derived from refractory/relapse AML (R/R AML) patients who relapsed from prior treatment with Venetoclax plus HMA.

Examples Part 1- Preparation of the compounds of formula (I)

Representative compounds formula (I) are summarized in the table 1 below:

Table 1

Representative compounds of formula (I) can be synthesized according to the following procedures.

General analytical procedures The ¹H and ¹³C NMR spectra were recorded on a Bruker Advance ALS300 and DRX400 MHz from Bruker. Chemical shifts are reported in ppm (5) and were referenced to DMSO- d6 (¹H, 2.50 ppm; ¹³C, 39.52 ppm) or CDCI3 (7.26 ppm). The coupling constants (J) were given in Hz.

The HRMS-ESI mass spectra were recorded in positive-ion mode on a hybrid quadrupole time-of-flight mass spectrometer (MicroTOFQ-ll, Bruker Daltonics, Bremen) with an Electrospray Ionization (ESI) ion source. For the mass spectrometry of low resolution, LRMS-ESI mass spectra were recorded in a Thermo Finnigan MAT 95 XL spectrometer.

Example 1 : S-methyl 4-r2-ethoxyethyl(methyl)aminol-4-methyl-pent-2-vnethioate Preparation of N-(2-ethoxyethyl)-N,2-dimethyl-but-3-vn-2-amine : To a solution of N- methyl-N-(2’hydroxyethyl)-3-amino-3methyl-1 -butyne (Easton, Nelson R.; Hennion, George F. U.S. (1967), US 3337625 19670822.) (1.0 g, 7.08 mmol) and iodoethane (0.98 mL, 7.6 mmol) in THF (12 mL) was added NaH (0.459 g, 11.5 mmol) at room temperature and the mixture was refluxed for 3 h. Mixture was then carefully hydrolyzed at room temperature by water and extracted by EtOAc (3x25 mL). Combined organic layers were washed with brine, dried over Na₂SO₄ and concentrated in vacuo. Purification of the crude by chromatography on silicagel (petroleum ether/EtOAc=70/30) gave pure N-(2- ethoxyethyl)-N,2-dimethyl-but-3-yn-2-amine (0.479 g, 40%) .

¹H NMR (300 MHz, DMSO) 5 3.45 - 3.36 (m, 4H), 3.11 (s, 1 H), 2.51 (t, J = 6.7 Hz,. 2H), 2.20 (s, 3H), 1.27 (s, 6H), 1.09 (t, J = 7.0 Hz, 3H).

ESI-LRMS 170.0 [M+HJ+.

Preparation of S-methyl 4-[2-ethoxyethyl(methyl)amino]-4-methyl-pent-2-ynethioate : To N-(2-ethoxyethyl)-N,2-dimethyl-but-3-yn-2-amine (0.367 g, 2.17 mmol) in THF (11 mL) was added dropwise a 2.28 M n-BuLi solution in hexane (1.14 mL, 2.60 mmol) at -70°C. After 5 min at -70°C the reaction mixture was warmed to 0°C, maintained 10 min at this temperature then cooled at -70°C before a 30 min bubbling with carbonyl sulfide (COS) through the solution. The yellow solution was warmed to 0°C, stirred for additional 10 min at this temperature before dropwise addition of iodomethane (0.162 mL, 2.60 mmol). The mixture was stirred for 2 h, carefully hydrolyzed at 0°C by water and extracted with ether. Combined organic layers were washed with brine, dried over Na₂SO₄ and concentrated in vacuo. Purification of the crude by chromatography on silicagel (petroleum ether/EtOAc=90/10) gave pure S-methyl 4-[2-ethoxyethyl(methyl)amino]-4-methyl-pent-2- ynethioate (0.369 g, 70%) as an near colorless oil.

¹H NMR (300 MHz, DMSO) 5 3.42 (t, J = 6.3 Hz, 2H), 3.42 (q, J = 7.0 Hz, 2H), 2.56 (t, J = 6.3 Hz, 2H), 2.39 (s, 3H), 2.25 (s, 3H), 1.36 (s, 6H), 1.10 (t, J = 7.0 Hz, 3H). ESI- HRMS calc for C₁₂H22NO₂S [M+H]+: 244.1366, found: 244.1362.

Example 2: S-methyl 4-r2-allyloxyethyl(methyl)aminol-4-methyl-pent-2-vnethioate

Preparation of N-(2-allyloxyethyl)-N,2-dimethyl-but-3-vn-2-amine : To N-methyl-N- (2’hydroxyethyl)-3-amino-3methyl-1-butyne (Easton, Nelson R.; Hennion, George F. U.S. (1967), US 3337625 19670822.)) (1 .0 g, 7.08 mmol) in THF (12 mL) was added NaH (0.340 g, 8.50 mmol) at 0°C. After 15 min at 0°C and 15 min at room temperature, n-Bu₄NI (0.026 g, 0.071 mmol) was added in one portion at 0°C followed by dropwise addition of allyl bromide (0.735 mL, 8.50 mmol). Reaction mixture was allowed to reach room temperature, stirred overnight, then carefully hydrolyzed by water and extracted by ether (3x25 mL). Combined organic layers were washed with brine (25 mL), dried over Na₂SO₄ and concentrated in vacuo. Purification by chromatography on silicagel (petroleum ether/ether=80/20 to 70/30) gave pure N-(2-allyloxyethyl)-N,2-dimethyl-but-3-yn-2-amine (0.941 g, 73%) as an oil.

¹H NMR (300 MHz, DMSO) 55.88 (ddt, J = 17.3, 10.5, 5.3 Hz, 1 H), 5.24 (ddd, J = 17.3, 3.8, 1.7 Hz, 1 H), 5.16 - 5.09 (m, 1 H), 3.93 (dt, J = 5.3, 1.6 Hz, 2H), 3.43 (t, J = 6.4 Hz, 2H), 3.12 (s, 1 H), 2.55 (t, J = 6.4 Hz, 2H), 2.21 (s, 3H), 1.27 (s, 6H).

ESI-LRMS 182.0 [M+HJ+.

Preparation of S-methyl 4-[2-allyloxyethyl(methyl)amino]-4-methyl-pent-2- ynethioate: The compound is obtained by using the same process as the one described for S-methyl 4-[2-ethoxyethyl(methyl)amino]-4-methyl-pent-2-ynethioate [example 1] starting from N-(2-allyloxyethyl)-N,2-dimethyl-but-3-yn-2-amine. Scale : 2.2 mmol. Purification by chromatography on silicagel (petroleum ether/EtOAc=90/10 to 80/20). Yield : 65%. Near colorless oil.

¹H NMR (300 MHz, DMSO) 55.88 (ddt, J = 17.3, 10.5, 5.3 Hz, 1 H), 5.24 (ddd, J = 17.3, 3.8, 1.7 Hz, 1 H), 5.17 - 5.10 (m, 1 H), 3.94 (dt, J = 5.3, 1.5 Hz, 2H), 3.45 (t, J = 6.2 Hz, 2H), 2.58 (t, J = 6.2 Hz, 2H), 2.39 (s, 3H), 2.26 (s, 3H), 1.36 (s, 6H).

ESI- HRMS calc for C₁₃H₂₂NO₂S [M+H]+: 256.1366, found: 256.1364.

Example 3: S-methyl 4-r2-benzyloxyethyl(methyl)aminol-4-methyl-pent-2-vnethioate Preparation of N-(2-benzyloxyethyl)-N,2-dimethyl-but-3-vn-2-amine : The compound is obtained by using the same process as the one described for N-(2-allyloxyethyl)-N,2- dimethyl-but-3-yn-2-amine [example 2] using 1.015 eq of NaH and 1.01 eq. of benzyl bromide. Purification by chromatography on silicagel (petroleum ether/EtOAc=90/10). Yield : 81%. Colorless oil. ¹H NMR (300 MHz, DMSO) 5 7.39 - 7.24 (m, 5H), 4.47 (s, 2H), 3.49 (t, J = 6.3 Hz, 2H), 3.12 (s, 1 H), 2.58 (t, J = 6.3 Hz, 2H), 2.21 (s, 3H), 1.27 (s, 6H). ESI-LRMS 232.0 [M+HJ+.

Preparation of S-methyl 4-[2-benzyloxyethyl(methyl)amino]-4-methyl-pent-2- ynethioate : The compound is obtained by using the same process as the one described for S-methyl 4-[2-ethoxyethyl(methyl)amino]-4-methyl-pent-2-ynethioate [example 1] starting from N-(2-benzyloxyethyl)-N,2-dimethyl-but-3-yn-2-amine. Scale : 2.2 mmol. Purification by chromatography on silicagel (petroleum ether/EtOAc=90/10). Yield : 79%. Colorless oil.

¹H NMR (300 MHz, DMSO) 5 7.37 - 7.26 (m, 5H), 4.48 (s, 2H), 3.52 (t, J = 6.1 Hz, 2H), 2.62 (t, J = 6.1 Hz, 2H), 2.38 (s, 3H), 2.26 (s, 3H), 1.36 (s, 6H).

ESI- HRMS calc for C₁₇H24NO₂S [M+H]+: 306.1522, found: 306.1514.

Alternative protocol: To N-(2-benzyloxyethyl)-N,2-dimethyl-but-3-yn-2-amine (0.650 g, 2.81 mmol) in THF (8 mL) was added dropwise a 2.28 M n-BuLi solution in hexane (1.36 mL, 3.09 mmol) at -70°C. After 5 min at -70°C the reaction mixture was warmed to 0°C, maintained 30 min at this temperature and CO₂ was bubbled through the solution for 30 min. The mixture was warmed to room temperature within 5 min then re-cooled at 0°C. Isobutyl chloroformate (0.40 ml, 3.08 mmol) was added dropwise and the mixture stirred for 10 min before addition of sodium methoxide (0.236 g, 3.37 mmol) in one portion. The mixture was warmed to room temperature stirred for additional 15 min at this temperature then carefully hydrolyzed at 0°C by water and extracted with ether. Combined organic layers were washed with brine, dried over Na₂SO₄ and concentrated in vacuo. Purification of the crude by chromatography on silicagel (petroleum ether/EtOAc=90/10) gave pure S-methyl 4-[2-ethoxyethyl(methyl)amino]-4-methyl-pent-2-ynethioate (0.307 g, 36%).

Example 4: S-methyl 4-methyl-4-rmethyl-r2-(m-tolylmethoxy)ethyllaminolpent-2- ynethioate

Preparation of N-2-dimethyl-N-[2-(m-tolylmethoxy)ethyl]but-3-yn-2-amine: The compound is obtained by using the same process as the one described for N-(2- allyloxyethyl)-N,2-dimethyl-but-3-yn-2-amine [example 2] using 3-Methylbenzyl bromide. Purification by chromatography on silicagel (petroleum ether/EtOAc=90/10). Scale : 4.5 mmol. Yield : 79%. Colorless oil.

¹H NMR (300 MHz, DMSO) 5 7.26 - 7.19 (m, 1 H), 7.16 - 7.05 (m, 3H), 4.43 (s, 2H), 3.48 (t, J = 6.3 Hz, 2H), 3.12 (s, 1 H), 2.57 (t, J = 6.3 Hz, 2H), 2.30 (s, 3H), 2.21 (s, 3H), 1.27 (s, 6H).

ESI-LRMS 246.1 [M+HJ+. Preparation of S-methyl 4-methyl-4-[methyl-[2-(m-tolylmethoxy)ethyl]amino]pent-2- ynethioate : The compound is obtained by using the same process as the one described for S-methyl 4-[2-ethoxyethyl(methyl)amino]-4-methyl-pent-2-ynethioate [example 1] starting from N-2-dimethyl-N-[2-(m-tolylmethoxy)ethyl]but-3-yn-2-amine. Scale : 1.3 mmol. Purification by chromatography on silicagel (petroleum ether/EtOAc=90/10). Yield : 77%. Colorless oil.

¹H NMR (300 MHz, DMSO) 5 7.26 - 7.19 (m, 1H), 7.16 - 7.05 (m, 3H), 4.44 (s, 2H), 3.50 (t, J = 6.1 Hz, 2H), 2.62 (t, J = 6.1 Hz, 2H), 2.38 (s, 3H), 2.30 (s, 3H), 2.26 (s, 3H), 1.36 (s, 6H).

ESI- HRMS calc for C₁₈H₂6NO₂S [M+H]+: 320.1679, found: 320.1667.

Example 5: S-methyl 4-r2-r(3,4-dimethvlphenvl)methoxvlethvl-methvl-aminol-4- methyl-pent-2-ynethioate

Preparation of N-[2-[(3,4-dimethylphenyl)methoxy]ethyl]-N,2-dimethyl-but-3-yn-2- amine : The compound is obtained by using the same process as the one described for N- (2-allyloxyethyl)-N,2-dimethyl-but-3-yn-2-amine [example 2] using 3,4-Dimethylbenzyl bromide. Purification by chromatography on silicagel (petroleum ether/EtOAc=60/40). Scale : 3.8 mmol. Yield : 60%. Colorless oil.

¹H NMR (300 MHz, DMSO) 5 7.12 - 7.06 (m, 2H), 7.04 - 6.99 (m, 1 H), 4.39 (s, 2H), 3.45 (t, J = 6.3 Hz, 2H), 3.12 (s, 1H), 2.56 (t, J = 6.3 Hz, 2H), 2.20 (s, 6H), 2.19 (s, 3H), 1.27 (s, 6H).

ESI-LRMS 182.0 [M+H]+. ESI-LRMS 260.0 [M+HJ+.

Preparation of S-methyl 4-[2-[(3,4-dimethylphenyl)methoxy]ethyl-methyl-amino]-4- methyl-pent-2-ynethioate : The compound is obtained by using the same process as the one described for S-methyl 4-[2-ethoxyethyl(methyl)amino]-4-methyl-pent-2-ynethioate [example 1] starting from N-[2-[(3,4-dimethylphenyl)methoxy]ethyl]-N,2-dimethyl-but-3-yn- 2-amine. Scale: 1.3 mmol. Purification by chromatography on silicagel (petroleum ether/EtOAc=90/10). Yield : 77%. Near colorless oil.

¹H NMR (300 MHz, DMSO) 5 7.12 - 7.06 (m, 2H), 7.05 - 6.99 (m, 1 H), 4.40 (s, 2H), 3.48 (t, J = 6.2 Hz, 2H), 2.60 (t, J = 6.2 Hz, 2H), 2.38 (s, 3H), 2.25 (s, 3H), 2.20 (s, 3H), 2.19 (s, 3H), 1.36 (s, 6H).

ESI- HRMS calc for C₁₉H₂₈NO₂S [M+H]+: 334.1835, found: 334.1825. Preparation of N-[2-[(4-methoxyphenyl)methoxy]ethyl]-N,2-dimethyl-but-3-yn-2- amine: The compound is obtained by using the same process as the one described for N- (2-allyloxyethyl)-N,2-dimethyl-but-3-yn-2-amine [example 2] using 1-(Bromomethyl)-4- methoxybenzene. Purification by chromatography on silicagel (DCM/MeOH=99/1 to 97.5/2.5). Scale : 4.0 mmol. Yield : 53%. Colorless oil.

1 H NMR (300 MHz, DMSO) 5 7.27 - 7.20 (m, 2H), 6.93 - 6.86 (m, 2H), 4.39 (s, 2H), 3.74 (s, 3H), 3.45 (t, J = 6.3 Hz, 2H), 3.12 (s, 1 H), 2.55 (t, J = 6.4 Hz, 2H), 2.20 (s, 3H), 1.27 (s, 6H).

ESI-LRMS 261.9 [M+HJ+.

Preparation of S-methyl 4-[2-[(4-methoxyphenyl)methoxy]ethyl-methyl-amino]-4- methyl-pent-2-ynethioate: The compound is obtained by using the same process as the one described for S-methyl 4-[2-ethoxyethyl(methyl)amino]-4-methyl-pent-2-ynethioate [example 1] starting from N-[2-[(4-methoxyphenyl)methoxy]ethyl]-N,2-dimethyl-but-3-yn-2- amine. Scale: 1.3 mmol. Purification by chromatography on silicagel (petroleum ether/EtOAc=80/20). Yield : 74%. Near colorless oil.

¹H NMR (300 MHz, DMSO) 5 7.27 - 7.21 (m, 2H), 6.93 - 6.87 (m, 2H), 4.40 (s, 2H), 3.74 (s, 3H), 3.48 (t, J = 6.2 Hz, 2H), 2.60 (t, J = 6.2 Hz, 2H), 2.38 (s, 3H), 2.25 (s, 3H), 1.36 (s, 6H).

ESI- HRMS calc for C₁₈H₂6NO₃S [M+H]+: 336.1628, found: 336.1613.

Example 7: S-methvI 4-r2-K3,4-dimethoxvphenvl)methoxvlethvl-methvl-aminol-4- methvl-pent-2-vnethioate

Preparation of N-[2-[(3,4-dimethoxyphenyl)methoxy]ethyl]-N,2-dimethyl-but-3-yn-2- amine: The compound is obtained by using the same process as the one described for N- (2-allyloxyethyl)-N,2-dimethyl-but-3-yn-2-amine [example 2] using 4-(Bromomethyl)-1,2- dimethoxybenzene. Purification by chromatography on silicagel (DCM/MeOH=99/1 to 97.5/2.5). Scale : 4.0 mmol. Yield : 67%. Colorless oil.

¹H NMR (300 MHz, DMSO) 5 6.94 - 6.88 (m, 2H), 6.87 - 6.80 (m, 1 H), 4.39 (s, 2H), 3.74 (s, 3H), 3.73 (s, 3H), 3.46 (t, J = 6.3 Hz, 2H), 3.12 (s, 1 H), 2.56 (t, J = 6.3 Hz, 2H), 2.21 (s, 3H), 1.27 (s, 6H).

ESI-LRMS 292.0 [M+HJ+.

Preparation of S-methyl 4-[2-[(3,4-dimethoxyphenyl)methoxy]ethyl-methyl-amino]-4- methyl-pent-2-ynethioate: The compound is obtained by using the same process as the one described for S-methyl 4-[2-ethoxyethyl(methyl)amino]-4-methyl-pent-2-ynethioate [example 1] starting from N-[2-[(3,4-dimethoxyphenyl)methoxy]ethyl]-N,2-dimethyl-but-3- yn-2-amine. Scale: 1.0 mmol. Purification by chromatography on silicagel (petroleum ether/EtOAc=90/10). Yield : 67%. Near colorless oil.

¹H NMR (300 MHz, DMSO) 5 6.94 - 6.87 (m, 1 H), 6.87 - 6.81 (m, 1 H), 4.40 (s, 2H), 3.74 (s, 3H), 3.73 (s, 3H), 3.48 (t, J = 6.1 Hz, 2H), 2.61 (t, J = 6.2 Hz, 2H), 2.38 (s, 3H), 2.26 (s, 3H), 1.36 (s, 6H).

ESI- HRMS calc for C19H28NO4S [M+H]+: 366.1734, found: 336.1720.

Example 8: S-methvI 4-r2-K3-chlorophenvl)methoxvlethvl-methvl-aminol-4-methvl- pent-2-ynethioate

Preparation of N-[2-[(3-chlorophenyl)methoxy]ethyl]-N,2-dimethyl-but-3-yn-2-amine: The compound is obtained by using the same process as the one described for N-(2- allyloxyethyl)-N,2-dimethyl-but-3-yn-2-amine [example 2] using 3-Chlorobenzyl bromide. Purification by chromatography on silicagel (petroleum ether/EtOAc=70/30). Scale : 4.0 mmol. Yield : 71 %. Colorless oil.

¹H NMR (300 MHz, DMSO) 5 7.43 - 7.25 (m, 4H), 4.49 (s, 2H), 3.50 (t, J = 6.2 Hz, 2H), 3.12 (s, 1 H), 2.58 (t, J = 6.2 Hz, 2H), 2.22 (s, 3H), 1.28 (s, 6H).

ESI-LRMS 266.0 [M+H]+.

Preparation of S-methyl 4-[2-[(3-chlorophenyl)methoxy]ethyl-methyl-amino]-4- methyl-pent-2-ynethioate: The compound is obtained by using the same process as the one described for S-methyl 4-[2-ethoxyethyl(methyl)amino]-4-methyl-pent-2-ynethioate [example 1] starting from N-[2-[(3-chlorophenyl)methoxy]ethyl]-N,2-dimethyl-but-3-yn-2- amine except that the reaction mixture was maintained at -70°C after n-BuLi addition for 30 min before COS bubbling. Scale : 1.3 mmol. Purification by chromatography on silicagel (petroleum ether/EtOAc=75/25). Yield : 63%. Near colorless oil.

¹H NMR (300 MHz, DMSO) 5 7.44 - 7.24 (m, 4H), 4.50 (s, 2H), 3.52 (t, J = 6.0 Hz, 2H), 2.63 (t, J = 6.0 Hz, 2H), 2.38 (s, 3H), 2.27 (s, 3H), 1.37 (s, 6H).

ESI- HRMS calc for C₁₇H23CINO₂S [M+H]+: 340.1133, found: 340.1120.

Preparation of N-[2-[(3-fluorophenyl)methoxy]ethyl]-N,2-dimethyl-but-3-yn-2-amine : The compound is obtained by using the same process as the one described for N-(2- allyloxyethyl)-N,2-dimethyl-but-3-yn-2-amine [example 2] using 3-fluorobenzyl bromide. Purification by chromatography on silicagel (petroleum ether/EtOAc=80/20). Scale : 4.0 mmol. Yield : 71 %. Colorless oil. Preparation of S-methyl 4-[2-[(3-fluorophenyl)methoxy]ethyl-methyl-amino]-4- methyl-pent-2-ynethioate: The compound is obtained by using the same process as the one described for S-methyl 4-[2-ethoxyethyl(methyl)amino]-4-methyl-pent-2-ynethioate [example 1] starting from N-[2-[(3-fluorophenyl)methoxy]ethyl]-N,2-dimethyl-but-3-yn-2- amine except that the reaction mixture was maintained at -70°C after n-BuLi addition for 30 min before COS bubbling. Scale : 1.3 mmol. Purification by chromatography on silicagel (petroleum ether/EtOAc=70/30). Yield : 87%. Near colorless oil.

¹H NMR (300 MHz, DMSO) 5 7.44 - 7.34 (m, 1 H), 7.20 - 7.05 (m, 3H), 4.51 (s, 2H), 3.53 (t, J = 6.1 Hz, 2H), 2.63 (t, J = 6.1 Hz, 2H), 2.38 (s, 3H), 2.27 (s, 3H), 1.37 (s, 6H).

ESI- HRMS calc for C₁₇H23FNO₂S [M+H]+: 324.1428, found: 324.1415

Example 10: S-methyl 4-methyl-4-rmethyl-r2-(2-pyridylmethoxy)ethyllamino]pent-2- ynethioate

Preparation of N-2-dimethyl-N-[2-(2-pyridylmethoxy)ethyl]but-3-yn-2-amine : The compound is obtained by using the same process as the one described for N-(2- allyloxyethyl)-N,2-dimethyl-but-3-yn-2-amine [example 2] starting from 2- (Bromomethyl)pyridine hydrobromide and using 4 eq of NaH. Purification by chromatography on silicagel (DCM/MeOH=99/1 to 95/5). Scale : 2.1 mmol. Yield : 71%. Yellow oil.

¹H NMR (300 MHz, DMSO) 5 8.50 (ddd, J = 4.8, 1.8, 0.9 Hz, 1 H), 7.80 (td, J = 7.7, 1.8 Hz, 1 H), 7.44 (d, J = 7.8 Hz, 1 H), 7.32 - 7.24 (m, 1 H), 4.55 (s, 2H), 3.56 (t, J = 6.2 Hz, 2H), 3.13 (s, 1 H), 2.61 (t, J = 6.2 Hz, 2H), 2.23 (s, 3H), 1.28 (s, 6H).

ESI-LRMS 233.1 [M+HJ+.

Preparation of S-methyl 4-methyl-4-[methyl-[2-(2-pyridylmethoxy)ethyl]amino]pent- 2-ynethioate: The compound is obtained by using the same processas the one described for S-methyl 4-[2-ethoxyethyl(methyl)amino]-4-methyl-pent-2-ynethioate [example 1] starting from N-2-dimethyl-N-[2-(2-pyridylmethoxy)ethyl]but-3-yn-2-amine and using 1.5 eq of n-BuLi, 1.5 eq of Mel and DCM extractions. Scale : 0.9 mmol. Purification by chromatography on silicagel (DCM/MeOH=99/1 to 90/10). Yield : 18%. Yellow oil.

¹H NMR (300 MHz, DMSO) 5 8.54 - 8.48 (m, 1 H), 7.80 (td, J = 7.7, 1.8 Hz, 1 H), 7.44 (d, J = 7.8 Hz, 1 H), 7.28 (dd, J = 6.7, 5.1 Hz, 1 H), 4.56 (s, 2H), 3.59 (t, J = 6.1 Hz, 2H), 2.65 (t, J = 6.1 Hz, 2H), 2.38 (s, 3H), 2.28 (s, 3H), 1.37 (s, 6H). ).

ESI- HRMS calc for C₁₆H₂3N₂O2S [M+H]+: 3071475, found: 307.1471.

Example 11 : S-methyl 4-methyl-4-rmethyl-r2-(3-pyridylmethoxy)ethyllamino]pent-2- ynethioate Preparation of N-2-dimethyl-N-[2-(3-pyridylmethoxy)ethyl]but-3-yn-2-amine : The compound is obtained by using the same process as the one described for N-(2- allyloxyethyl)-N,2-dimethyl-but-3-yn-2-amine [example 2] starting from 3- (Bromomethyl)pyridine hydrobromide and using 4 eq of NaH. Purification by chromatography on silicagel (DCM/MeOH=99/1 to 95/5). Scale : 2.1 mmol. Yield : 67%. Yellow oil.

¹H NMR (300 MHz, DMSO) 5 8.56 - 8.52 (m, 1H), 8.49 (dd, J = 4.8, 1.7 Hz, 1 H), 7.78 - 7.70 (m, 1 H), 7.38 (ddd, J = 7.8, 4.8, 0.8 Hz, 1 H), 4.52 (s, 2H), 3.52 (t, J = 6.2 Hz, 2H), 3.12 (s, 1 H), 2.58 (t, J = 6.2 Hz, 2H), 2.21 (s, 3H), 1.27 (s, 6H).

ESI-LRMS 233.1 [M+H]⁺.

Preparation of S-methyl 4-methyl-4-[methyl-[2-(3-pyridylmethoxy)ethyl]amino]pent- 2-ynethioate: The compound is obtained by using the same process as the one described for S-methyl 4-[2-ethoxyethyl(methyl)amino]-4-methyl-pent-2-ynethioate [example 1] starting from N-2-dimethyl-N-[2-(3-pyridylmethoxy)ethyl]but-3-yn-2-amine and using 1.5 eq of n-BuLi,1.5 eq of Mel and DCM extractions. Scale : 0.4 mmol. Purification by chromatography on silicagel (DCM/MeOH=99/1 to 95/5). Yield : 15%. Yellow oil.

¹H NMR (300 MHz, DMSO) 5 8.54 (d, J = 1.5 Hz, 1 H), 8.49 (dd, J = 4.8, 1.7 Hz, 1 H), 7.78 - 7.70 (m, 1 H), 7.38 (ddd, J = 7.8, 4.8, 0.8 Hz, 1 H), 4.53 (s, 2H), 3.54 (t, J = 6.1 Hz, 2H), 2.63 (t, J = 6.1 Hz, 2H), 2.38 (s, 3H), 2.26 (s, 3H), 1.36 (s, 6H) ).

ESI- HRMS calc for C₁₆H₂3N₂O2S [M+H]+: 3071475, found: 307.1474.

Example 12: S-methyl 4-methyl-4-rmethyl-r2-(4-pyridylmethoxy)ethyllaminolpent-2- ynethioate

Preparation of N-2-dimethyl-N-[2-(4-pyridylmethoxy)ethyl]but-3-yn-2-amine : The compound is obtained by using the same process as the one described for N-(2- allyloxyethyl)-N,2-dimethyl-but-3-yn-2-amine [example 2] starting from 4- (Bromomethyl)pyridine hydrobromide and using 4 eq of NaH. Purification by chromatography on silicagel (DCM/MeOH=99/1 to 95/5). Scale : 2.1 mmol. Yield : 95%. Yellow oil.

¹H NMR (300 MHz, DMSO) 5 8.56 - 8.49 (m, 2H), 7.38 - 7.27 (m, 2H), 4.54 (s, 2H), 3.53 (t, J = 6.2 Hz, 2H), 3.13 (s, 1H), 2.61 (t, J = 6.2 Hz, 2H), 2.23 (s, 3H), 1.28 (s, 6H). ESI-LRMS 233.1 [M+HJ+.

Preparation of S-methyl 4-methyl-4-[methyl-[2-(4-pyridylmethoxy)ethyl]amino]pent- 2-ynethioate: The compound is obtained by using the same process as the one described for S-methyl 4-[2-ethoxyethyl(methyl)amino]-4-methyl-pent-2-ynethioate [example 1] starting from N-2-dimethyl-N-[2-(4-pyridylmethoxy)ethyl]but-3-yn-2-amine and using 1.5 eq of n-BuLi,1.5 eq of Mel and DCM extractions. Scale : 1.0 mmol. Purification by chromatography on silicagel (DCM/MeOH=99/1 to 95/15). Yield : 24%. Yellow oil.

¹H NMR (300 MHz, DMSO) 5 8.60 - 8.45 (m, 2H), 7.37 - 7.26 (m, 2H), 4.55 (s, 2H), 3.56 (t, J = 6.0 Hz, 2H), 2.65 (t, J = 6.0 Hz, 2H), 2.38 (s, 3H), 2.28 (s, 3H), 1.37 (s, 6H).

ESI- HRMS calc for C₁₆H₂3N₂O2S [M+H]+: 3071475, found: 307.1470.

Example 13: S-methyl 4-((4-(benzyloxy)butyl)(methyl)amino)-4-methylpent-2- ynethioate

Preparation of 4-(methyl(2-methylbut-3-yn-2-yl)amino)butan-1-ol : This compound was prepared by standard protocols previously described for the synthesis of N-methyl-N- (2’hydroxyethyl)-3-amino-3methyl-1-butyne (Easton, Nelson R.; Hennion, George F. U.S. (1967), US 3337625 19670822.) starting from commercially available 4- (methylamino)butan-l-ol. 4-(methyl(2-methylbut-3-yn-2-yl)amino)butan-1-ol was obtained as a bright yellow oil. Scale 3 mmol. Yield : 99%.

¹H NMR (300MHz, DMSO) 5 4.41 (t, J = 5.2Hz, 1 H), 3.42 - 3.33 (m, 2H), 3.09 (s, 1 H), 2.38 - 2.29 (m, 2H), 2.14 (s, 3H), 1.45 - 1.36 (m, 4H), 1.27 (s, 6H).

ESI - LRMS : 170.1 [M+H]+

Preparation of N-(4-(benzyloxy)butyl)-N,2-dimethylbut-3-yn-2-amine : The compound was obtained by using the same process as the one described for N-(2-allyloxyethyl)-N,2- dimethyl-but-3-yn-2-amine [example 2] starting from 4-(methyl(2-methylbut-3-yn-2- yl)amino)butan-1-ol and using 1.015 eq of NaH and 1.01 eq. of benzyl bromide. Purification by chromatography on silicagel (cyclohexane/EtOAc=80/20). Scale 2.4 mmol. Yield : 55%. Yellow oil.

¹H NMR (300MHz, DMSO) 5 7.39 - 7.22 (m, 5H), 4.44 (s, 2H), 3.42 (t, J = 6.3Hz, 2H), 3.08 (s, 1 H), 2.34 (t, J = 6.9Hz, 2H), 2.13 (s, 3H), 1.60 - 1.37 (m, 4H), 1.26 (s, 6H).

ESI - LRMS : 260.2 [M+H]+

Preparation of S-methyl 4-((4-(benzyloxy)butyl)(methyl)amino)-4-methylpent-2- ynethioate : The compound was obtained by using the same process as the one described for S-methyl 4-[2-ethoxyethyl(methyl)amino]-4-methyl-pent-2-ynethioate [example 1] starting from N-(4-benzyloxybutyl)-N,2-dimethyl-but-3-yn-2-amine Purification by chromatography on silicagel (cyclohexane/EtOAc= 90/10). Scale 0.77 mmol. Yield : 80%. Yellow oil.

1 H NMR (300MHz, CDCI3) 5 7.38 - 7.19 (m, 5H), 4.48 (s, 2H), 3.47 (t, J = 6.1 Hz, 2H), 2.45 (t, J = 6.9Hz, 2H), 2.35 (s, 3H), 2.26 (s, 3H), 1 .73 - 1 .48 (m, 4H), 1 .39 (s, 6H).

ESI - HRMS : calc for C19H28NO2S [M+H]+ 334.1835, found 334.1840. Example 14: S-methyl 4-((2-hvdroxyethyl)(methyl)amino)-4-methylpent-2-vnethioate a) see Easton, Nelson R.; Hennion, George F. , U.S. (1966), US 3285913 b) 3,4-DHP, pTSA, DCM (70%) c) nBuLi, THF, -70°C, carbonyl sulfide then Mel 0°C (59%) d) pTSA, MeOH, room temperature (90%)

Preparation of Compound 3: N,2-dimethyl-N-(2-((tetrahydro-2H-pyran-2- yl)oxy)ethyl)but-3-yn-2-amine: To 2-(methyl(2-methylbut-3-yn-2-yl)amino)ethanol 2 (3.00 g, 21.2 mmol) and 3,4-Dihydro-2H-pyran (5.0 eq) in anhydrous DCM (135 mL) was added p-toluenesulfonic acid (0.1 eq) at room temperature. The reaction mixture was stirred overnight, washed with aqueous saturated NaHCO₃ (30 mL) then brine (30 mL). The organic layer was dried over sodium sulfate, filtered and concentrated in vacuo. The residue was first purified by short-path distillation using Kugelrohr apparatus (10-12 Torrs, oven 155°C) then by flash chromatography on silica gel (petroleum ether/ethyl acetate 95/5 to 60/40) to give compound 3 as an oil (yield 70%).

¹H NMR (300 MHz, DMSO) 5 4.60 - 4.50 (m, 1 H), 3.82-3.70 (m, 1 H), 3.59-3.56 (m, 1 H), 3.49 - 3.35 (m, 2H), 3.12 (s, 1 H), 2.55 (t, J = 6.5 Hz, 2H), 2.21 (s, 3H), 1 .77 - 1.35 (m, 6H), 1.27 (s, 6H).

Preparation of Compound 4: S-methyl 4-methyl-4-(methyl(2-((tetrahydro-2H-pyran-2- yl)oxy) ethyl) amino) pent-2-ynethioate: To the acetylenic amine 3 (1.00 g, 4.44 mmol) in anhydrous THF (22 mL) was added n-Butyllithium solution (2.2 M in hexanes, 1.5 eq) dropwise. The mixture was allowed to reach to 0°C within 10 minutes then re-cooled to - 70°C before carbonyl sulfide bubbling. After 30 minutes the bright yellow solution was carefully warmed to 0°C, stirred 30 minutes at this temperature and methyl iodide (1.2 eq) was added dropwise. The reaction mixture was stirred for 2 hours at 0°C before hydrolysis by water. Extractive work-up by DCM (washing with brine, drying with sodium sulfate and concentration under reduced pressure) gave a crude which was purified by chromatography on silica gel (petroleum ether/ethyl acetate 90/10 to 60/40) to give compound 4 as an oil (yield 59%).

¹H NMR (300 MHz, DMSO) 54.58 (t, J = 3.2 Hz, 1 H), 3.75 (ddd, J = 11.4, 7.9, 3.3 Hz, 1 H), 3.70 - 3.60 (m, 1 H), 3.49 - 3.38 (m, 2H), 2.59 (t, J = 6.3 Hz, 2H), 2.39 (s, 3H), 2.27 (s, 3H), 1.77 - 1.39 (m, 6H), 1.36 (s, 6H). ¹³C NMR (75 MHz, CDCI₃) 5 176.62 (C=O), 98.94 (CH), 96.46 (C), 80.97 (C), 66.55 (CH₂), 62.37 (CH₂), 55.19 (C), 52.43 (CH₂), 37.92 (CH₃), 30.75 (CH₂), 28.01 (2XCH₃), 25.59 (CH₂), 19.63 (CH₂), 12.61 (CH₃).

Preparation of Compound 5: S-methyl 4-((2-hydroxyethyl)(methyl)amino)-4- methylpent-2-ynethioate: To the aminothiolester 4 (1.00 g, 3.34 mmol) in methanol (15 mL) was added p-toluenesulfonic acid (1.1 eq) at room temperature. The reaction mixture was stirred overnight, washed with aqueous saturated NaHCO₃ (30 mL) then brine (30 mL). The organic layer was dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (petroleum ether/ethyl acetate 80/20 to 20/80) to give compound 5 as an oil (yield 90%).

¹H NMR (300 MHz, DMSO) 5 4.42 (t, J = 5.6 Hz, 1 H), 3.44 (td, J = 6.7, 5.6 Hz, 2H), 2.46 (, J = 6.7 Hz, 2H), 2.39 (s, 3H), 2.24 (s, 3H), 1 .36 (s, 6H). ¹³C NMR (75 MHz, CDCI₃) 5 176.46 (C=O), 95.56 (C), 80.89 (C), 58.92 (CH₂), 54.99 (C) , 53.52 (CH₂), 36.12 (CH₃), 27.88 (2XCH₃), 12.53(CH₃). ESI-HRMS: Calc, for C₁₀H₁₈NO₂S [M+H]⁺ 216.1053 found 216.1043.

Example 15. S-methyl 4-methyl-4-rmethyl-r2-(2-naphthylmethoxy)ethyllamino]pent-2- ynethioate

Preparation of N,2-dimethyl-N-[2-(2-naphthylmethoxy)ethyl]but-3-yn-2-amine: The compound is obtained by using the same process as the one described for N-(2- allyloxyethyl)-N,2-dimethyl-but-3-yn-2-amine [example 2] using 1.015 eq of NaH and 1.01 eq. of 2-naphtyl bromide. Purification by chromatography on silicagel (petroleum ether/EtOAc=90/10). Yield: 61%. orange oil.

1 H NMR (300 MHz, DMSO) 5 7.91 - 7.88 (m, 4H), 7.50 - 7.48 (m, 3H), 4.65 (s, 2H), 3.55 (t, 6.1 Hz, 2H), 3.13 (s, 1 H), 2.62 (t, 6.2Hz, 2H), 2.23 (s, 3H), 1.28 (s, 6H).

Preparation of S-methyl 4-methyl-4-[methyl-[2-(2- naphthylmethoxy)ethyl]amino]pent-2-ynethioate: The compound is obtained by using the same process as the one described for S-methyl 4-[2-ethoxyethyl(methyl)amino]-4- methyl-pent-2-ynethioate [example 1] starting from N,2-dimethyl-N-[2-(2- naphthylmethoxy)ethyl]but-3-yn-2-amine. Scale: 0.65 mmol. Purification by chromatography on silicagel (petroleum ether/EtOAc=85/15). Yield: 28%. Yellow oil.

1 H NMR (300 MHz, DMSO) 57.95 - 7.82 (m, 4H), 7.55 - 7.41 (m, 3H), 4.66 (s, 2H), 3.57 (t, J = 6.2Hz, 2H), 2.66 (t, J = 6.1 Hz, 2H), 2.38 (s, 3H), 2.28 (s, 3H), 1.37 (s, 6H).

ESI - HRMS : calc, for C₂₁H₂₆NO₂S 356.1683, found 356.1679 [M+H]+

Example 16: S-methyl 4-methyl-4-rmethyl-r2-r(2,6,6-trimethylcyclohexen-1- yl)methoxylethyllamino]pent-2-vnethioate Preparation of N,2-dimethyl-N-[2-[(2,6,6-trimethylcyclohexen-1 -yl)methoxy]ethyl]but- 3-yn-2-amine: The compound is obtained by using the same process as the one described for N-(2-allyloxyethyl)-N,2-dimethyl-but-3-yn-2-amine [example 2] using 1.015 eq of NaH and 1.01 eq. of 2-(Bromomethyl)-1 ,3,3-trimethyl-1-cyclohexene (prepared from p- Cyclocitral by known protocols (WO 2015048363)). Purification by chromatography on silicagel (petroleum ether/EtOAc=95/05). Yield: 67%. Pale yellow oil.

3.86 (s, 2H), 3.41 (t, J = 6.5Hz, 2H), 3.12 (s, 1 H), 2.53 (t, J = 6.4Hz, 2H), 2.20 (s, 3H), 1.90 (t, J = 5.9Hz, 2H), 1.62 (s, 3H), 1.57 - 1.49 (m, 2H), 1.41 - 1 .33 (s, 2H), 1.27 (s, 6H), 0.97 (s, 6H).

Preparation of S S-methyl 4-methyl-4-[methyl-[2-[(2,6,6-trimethylcyclohexen-1- yl)methoxy]ethyl]amino]pent-2-ynethioate: The compound is obtained by using the same process as the one described for S-methyl 4-[2-ethoxyethyl(methyl)amino]-4-methyl- pent-2-ynethioate (example 15) starting from N,2-dimethyl-N-[2-[(2,6,6- trimethylcyclohexen-1-yl)methoxy]ethyl]but-3-yn-2-amine. Purification by chromatography on silicagel (petroleum ether/EtOAc=90/10). Yield: 63%. Yellow oil.

H NMR (300 MHz, DMSO) 5 3.87 (s, 2H), 3.44 (t, J = 6.3 Hz, 2H), 2.57 (t, J = 6.3 Hz, 2H), 2.38 (s, 3H), 2.26 (s, 3H), 1.91 (t, J = 6.2 Hz, 2H), 1.62 (s, 3H), 1.57 - 1.49 (m, 2H), 1.36 (s, 6H), 1.39 - 1 .34 (m, 2H), 0.97 (s, 6H).

ESI - HRMS calc for C20H34NO2S [M+H]+: 352.2305, found : 352.2289.

Example 17: S-methyl 2,5, 10,11, 11-pentamethyl-6-oxo-7-oxa-2,5,10-triazatetradec-12- yne-14-thioate

Preparation of compound 6: A solution of p-nitrochloroformate (140 mg, 0.70 mmol, 1.5 eq) in dichloromethane (DCM) (2.5 mL) is added dropwise to a solution of compound 5 of Example 14 (100 mg; 0.46 mmol) with triethylamine (TEA) (1.5 eq) in DCM (1.5 mL) at 0°C. After 10 minutes at 0°C, the reaction mixture is heated to room temperature with stirring until complete conversion (verification by TLC, 1 h). The mixture is then diluted in DCM (30 mL) then washed washed with brine (30 mL). The organic layer was dried over sodium sulfate, filtered and concentrated in vacuo. Purification by chromatography on silica gel (petroleum ether/ethyl acetate 100/70/30) to give compound 6 in the form of an amorphous solid (yield 82%).

1 H NMR (300MHz, DMSO): 5 (ppm) 8.35-8.30 (m, 2H), 7.59-7.53 (m, 2H), 4.30 (t, J = 5.7 Hz, 2H), 2.75 (t, J = 5.7 Hz , 2H), 2.39 (s, 3H), 2.30 (s, 3H), 1.39 (s, 6H).

Preparation of S-methyl 2,5,10,11,11-pentamethyl-6-oxo-7-oxa-2,5,10-triazatetradec- 12-yne-14-thioate: To compound 6 of example 18 (200 mg, 0.53 mmol) in 3 mL of DCM (3 mL) is added TEA (1 .2 eq) at room temperature. To the reaction mixture, a solution of le N, N, N'-trimethylethylenediamine (1.2 eq) in dichloromethane (2.3 mL) is added at 0 ° C. The bright yellow reaction mixture is warmed to room temperature and stirred overnight. The reaction mixture, a bright yellow color, was stirred overnight at room temperature, diluted with dichloromethane (40 mL) then washed with brine (30 mL). The organic layer was dried over sodium sulfate, filtered and concentrated in vacuo. Purification by chromatography on silica gel (petroleum ether / ethyl acetate 75/25 to 20/80). Yield: 70%. Oil.

Purification by chromatography on silica gel DCM/MeOH (85/15). Yield: 34%. Yellow oil. 1 H NMR (300 MHz, DMSO) 5 4.01 (t, J = 5.9Hz, 2H), 3.32-3.25 (m, 2H), 2.83 (large s, 3H), 2.62 (t, J = 5.8Hz, 2H), 2.6-2.5 (m partially hide by solvent peak, 2H), 2.39 (s, 3H), 2.27 (s,3H), 2.17 (large s, 6H), 1.36 (s, 6H).

ESI-HRMS calc for C16H30N3O3S [M+H]+: 344.1992, found : 344.1993.

Example 18: S-methyl 4-(((1R,2R)-2-(benzyloxy)cvclopentyl)(methyl)amino)-4- methylpent-2-ynethioate

Preparation of (1 R, 2R)-2-benzyloxy-N-(1,1-dimethylprop-2-ynyl)cyclopentanamine :

To a solution of commercially available (1 R,2R)-2-(benzyloxy)cyclopentanamine (0.93 g, 4.86 mmol), 3-chloro-3-methylbut-1-yne (1.3eq) and triethylamine (1.3eq) in THF (20 mL) was added Cui (8 mol%) at room temperature. The mixture was left to stir overnight. The solvent was evaporated under reduced pressure and the crude was then diluted in aqueous saturated NaHCO₃ solution, extracted with ethyl acetate. Combined organic layers were washed with 2% NH₄OH aqueous solution then brine, dried over Na₂SO₄ and the solvent evaporated under reduced pressure. (1 R, 2R)-2-benzyloxy-N-(1 ,1-dimethylprop-2- ynyl)cyclopentanamine was obtained as a brown oil. Yield : 99%.

¹H NMR (300 MHz, DMSO) 5 7.37 - 7.16 (m, 5H), 4.56 - 4.36 (m, 2H), 3.72 - 3.58 (m, 1 H), 3.28 - 3.19 (m, 1 H), 3.08 (s, 1 H), 2.00 - 1.88 (m, 1 H), 1.86 - 1.67 (m, 2H), 1.65 - 1.49 (m, 3H), 1.40 - 1.28 (m, 1 H), 1.25 (s, 3H), 1.25 (s, 3H).

Preparation of (1 R,2R)-2-(benzyloxy)-N-methyl-N-(2-methylbut-3-yn-2- yl)cyclopentanamine : To (1 R,2R)-2-(benzyloxy)-N-(2-methylbut-3-yn-2- yl)cyclopentanamine (0.25 g, 0.97 mmol) were added 5 eq of formic acid and 1.5 eq of formaldehyde (37% in water). The mixture was refluxed overnight then 2N HCI was added until pH 1 was reached and washed with ether. The aqueous layer was basified with 1 N NaOH, and extracted with DCM. The organic layer was washed with brine, dried over Na₂SO₄ and the solvents evaporated under reduced pressure. The crude is then purified by chromatography on silica gel (petroleum ether/EtOAc = 80/20), giving a yellow oil. Yield : 68%. ¹H NMR (300 MHz, DMSO) 6 7.43 - 7.17 (m, 5H), 4.52(m, 2H), 3.94 - 3.78 (m, 1 H), 3.62 - 3.49 (m, 1 H), 3.11 (s, 1 H), 2.20 (s, 3H), 1.78 - 1.45 (m, 6H), 1.37 (s, 3H), 1.34 (s, 3H). ESI - LRMS: 272.1 [M+H]⁺.

Preparation of S-methyl 4-(((1R,2R)-2-(benzyloxy)cyclopentyl)(methyl)amino)-4- methylpent-2-ynethioate : The compound was obtained by using the same process as the one described for S-methyl 4-[2-ethoxyethyl(methyl)amino]-4-methyl-pent-2-ynethioate [example 1] starting from (1 R,2R)-2-(benzyloxy)-N-methyl-N-(2-methylbut-3-yn-2- yl)cyclopentanamine. Purification by chromatography on silica gel (petroleum ether/EtOAc= 80/20 then DCM = 100). Scale 0.64 mmol. Yield : 47%. Yellow oil.

¹H NMR (300 MHz, DMSO) 5 7.35 - 7.20 (m, 5H), 4.51 (s, 2H), 3.92 - 3.80 (m, 1 H), 3.59 - 3.45 (m, 1 H), 2.36 (s, 3H), 2.24 (s, 3H), 1 .78 - 1 .49 (m, 6H), 1.45 (s, 3H), 1 .42 (s, 3H).

ESI - HRMS calc for C20H28NO2S [M+H]+: 346.1835, found: 346.1824.

Example 19: (S)-S-methyl 4-(2-((benzyloxy)methyl)pyrrolidin-1-yl)-4-methylpent-2- ynethioate:

Preparation of (S)-2-((benzyloxy)methyl)-1-(2-methylbut-3-yn-2-yl)pyrrolidine: The compound was obtained by using the same process as the one described for N-(2- allyloxyethyl)-N,2-dimethyl-but-3-yn-2-amine [example 2] starting from (S)-(1-(2-methylbut- 3-yn-2-yl)pyrrolidin-2-yl)methanol 1.015 eq of NaH and 1.01 eq. of benzyl bromide. Purification by chromatography on silicagel (dichloromethane/methanol = 95/5). Scale 3.0 mmol. Yield : 60%. Orange oil.

¹H NMR (300 MHz, DMSO) 5 7.41 - 7.19 (m, 5H), 4.46 (s, 2H), 3.24 (dd, J = 8.3, 2.7 Hz, 1 H), 3.14 (dd, J = 7.6, 3.2 Hz, 1 H), 3.11 - 3.02 (m, 2H), 2.86 (dd, J = 8.4, 6.0 Hz, 1 H), 2.65 - 2.54 (m, 1 H), 1.81 - 1.51 (m, 4H), 1.25 (s, 3H), 1.23 (s, 3H).

ESI-LRMS: 258.1 [M+HJ+.

Preparation of (S)-S-methyl 4-(2-((benzyloxy)methyl)pyrrolidin-1-yl)-4-methylpent-2- ynethioate: The compound was obtained by using the same process as the one described for S-methyl 4-[2-ethoxyethyl(methyl)amino]-4-methyl-pent-2-ynethioate [example 1] starting from (S)-2-((benzyloxy)methyl)-1-(2-methylbut-3-yn-2-yl)pyrrolidine. Purification by chromatography on silicagel (petroleum ether/EtOAc=80/20). Scale 1.2 mmol. Yield : 50%. Orange oil.

¹H NMR (300 MHz, DMSO) 5 7.46 - 7.18 (m, 5H), 4.47 (s, 2H), 3.29 - 3.23 (m, 1 H), 3.17 - 3.03 (m, 2H), 2.94 (dd, J = 8.5, 5.7 Hz, 1 H), 2.62 - 2.53 (m, 1 H), 2.38 (s, 3H), 1 .85 - 1.58 (m, 4H), 1.34 (s, 3H), 1.32 (s, 3H).

Example 20: (R)-S-methyl 4-(3-(benzyloxy)pyrrolidin-1-yl)-4-methylpent-2-vnethioate: Preparation of (R)-3-(benzyloxy)-1-(2-methylbut-3-yn-2-yl)pyrrolidine : The compound was obtained by using the same process as the one described for N-(2-allyloxyethyl)-N,2- dimethyl-but-3-yn-2-amine [example 2] starting from (R)-1-(2-methylbut-3-yn-2- yl)pyrrolidin-3-ol using 1.01 eq of NaH and 1.01 eq. of benzyl bromide. Purification by chromatography on silicagel (dichloromethane/methanol = 98/2). Scale 3.3 mmol. Yield : 69%. Orange oil.

¹H NMR (300 MHz, DMSO) 6 7.40 - 7.17 (m, 5H), 4.42 (s, 2H), 4.07 (tt, J = 7.3, 3.7Hz, 1 H) 3.13 (s, 1 H), 2.86 (dd, J = 9.6, 6.7 Hz, 1 H), 2.75 - 2.65 (m, 2H), 2.60 - 2.53 (m, 1 H), 2.06 - 1.95 (m, 1 H), 1.75 - 1.65 (m, 1 H), 1.29 (s, 6H).

ESI - LRMS 244.1 [M+HJ+.

Preparation of (R)-S-methyl 4-(3-(benzyloxy)pyrrolidin-1-yl)-4-methylpent-2- ynethioate: The compound was obtained by using the same process as the one described for S-methyl 4-[2-ethoxyethyl(methyl)amino]-4-methyl-pent-2-ynethioate [example 1] starting from (R)-3-(benzyloxy)-1-(2-methylbut-3-yn-2-yl)pyrrolidine. Purification by chromatography on silicagel (dichloromethane/methanol=98/2). Scale 1.2 mmol. Yield : 69%. Orange oil.

¹H NMR (300 MHz, DMSO) 5 7.32 (m, 5H), 4.44 (s, 2H), 4.09 (dq, J = 10.2, 3.5 Hz, 1 H), 2.97 - 2.83 (m, 1 H), 2.80 - 2.65 (m, 2H), 2.65 - 2.55 (m, 1 H), 2.37 (s, 3H), 2.11 - 1.90 (m, 1 H), 1.83 - 1.67 (m, 1 H), 1.37 (s, 6H).

ESI - HRMS calc for C₁₈H₂4NO₂S [M+H]+: 318.1522, found: 318.1518.

Example 21 : Preparation of S-methyl 4-dimethylamino-4-methyl-pent-2-vnethioate

To a solution of 0.855g of 3-dimethylamino-3-methyl-but-1-yne [HENNION, G.F., NELSON, K.W. (1957) J. Am. Chem. Soc., 79, 2142-2144] (2.80 mmol) in 14 ml of tetra hydrofuran, 4.07 ml (9.24 mmol) of a solution of 2.27 M denbutyllithium in hexane are added in 5 minutes at -70°C. After 5 minutes, the reaction medium is brought to 0°C and stirred for a further 30 minutes at this temperature. After returning to -70°C, 2 ml of carbon oxysulphide previously condensed are cannulated and the mixture is stirred for 30 minutes at -70°C. The reaction medium is then brought to 0°C over 30 minutes and then 0.575 ml (9.24 mmol) of methyl iodide is added and stirring is continued for 2 hours at 0°C. Diluted in 250 ml of ether, washed with saturated sodium chloride solution (3 x 30 ml), dried over sodium sulfate. After evaporation under vacuum and purification by chromatography on silica gel (elution with a petroleum ether/ethyl acetate mixture = 70/30), 1.16 g of the compound were isolated in the form of a colorless oil (yield: 81%).

¹H NMR (300 MHz, CDCI3): v = 1 .42 (s, 6H, (CH3) 2C), 2.31 (s, 6H, (CH3) 2N), 2.39 (s, 3H, CH3S). Example 22: Preparation of S-methyl 4-methyl-4-morpholin-4-yl-pent-2-ynethioate

The preparation is identical to that described in Example 21 using 3-di-n-propylamino-3- methyl-but-1-yne [HENNION, G.F., HANZEL, R.F., J. Am. Chem. Soc., 82, 4908-4912, (I960)] in place of 3-methyl-3-morpholin-4-yl-but-1-yne. Quantity: 1.5 mmol, purification by chromatography on silica gel (eluent petroleum ethe /ethyl acetate = 60/40), yield: 77%. Solid white.

¹H NMR (300 MHz, CDCI3): v = 1.43 (s, 6H, (CH3) 20), 2.40 (s, 3H, CH3S), 2.65 (m, 4H, CH2O), 3.97 (m, 4H, CH2N).

Example 23: Preparation of S methyl 4-methyl-4- (methyl (octyl) amino] pent-2- ynethioate

To N-methyl-N-(2-methylbu-3-yn-2-yl) hetan-1-amine (511 mg, 2.62 mmol) in solution in anhydrous THF (5 mL) is added a solution of nBuLi 2.28 M in hexane (1.26 mL, 1.1 eq) dropwise at -70°C. The reaction medium is brought to 0°C and CO₂ gas is introduced into the solution by bubbling through a needle for 30 min. The bubbling is continued for a further 5-10 min while the solution rises to room temperature. After returning to 0°C, isobutyl chloroformate (1.1 eq) is added dropwise. Stirring is continued during 15 min before adding MeSNa (1.2 eq) in one portion. The reaction medium is brought to room temperature and stirred for 15 min before hydrolysis with water. The treatment consists of extraction with ethyl acetate (x3) followed by washing the combined organic phases with a saturated aqueous solution of sodium chloride. After drying over Na₂SO₄ and concentrating on a rotary evaporator, the reaction crude is purified by chromatography on silica gel (petroleum ether/EtOAc = 95/5) to give the thiolester (445 mg). Yield: 63%. Solid white.

1 H NMR (300 MHz, DMSO) 5 2.38 (s, 3H), 2.37 (t, J = 7.0 Hz, 2H), 2.18 (s, 3H), 1.35 (s, 6H), 1.42 - 1.20 (m, 10H) , 0.86 (t, J = 6.7Hz, 3H). 13C NMR (75 MHz, CDCI3) 5 176.38 (C), 96.50 (C), 81.03 (C), 54.98 (C), 52.39 (CH2), 36.39, 31.85 (CH2), 29.25 (CH2), 28.79 (CH2) , 27.81 (2CH3), 27.41 (CH2), 22.62 (CH2), 14.09 (CH3), 12.41 (CH3). ESI-LRMS 270.2 [M + H] +.

Lipidic nanoparticle formulation of compounds

The aminothiolester compounds can be formulated as lipidic nanoparticles consisting of 2.5% of Lipoid S100 (Lipoid gmbh, Ludwigshafen, Germany), 20% Labrafac Lipophile WL1349 (Gattefosse, France), 17% Kolliphor HS15 (Sigma), 3% Sodium chloride (NaCI, (AppliChem), 60% of H20 and 12.2 mg/mL of an aminothiolester compound (e.g. DIMATE). The procedure was as follows: DI MATE (or another aminothiolester compound) was mixed with Labrafac and left steering until completely dissolved. In a separate falcon, Kolliphor, Lipoid, NaCI and the water were mixed. This last solution (acqueous phase) was transferred into the oily phase (DIMATE plus Labrafac) under steering, placed into a silicon oil bath (Silicon oil -50°C - +200°C, Sigma Aldrich) and heated up to 85°C under vigorous steering. Repeated cooling and heating steps were performed three times. At the end of the last heating step, the mixture was transferred into an ice bath and cooled down to 70°C. As soon as the formulation reached 70°C, cold water (4°C) was added to quench and the formulation was left in further agitation for 5 minutes.

Part 2: Use in combination with HMA

In these experiments, Dimate was not formulated in a nanoparticle.

Material and methods

Cell lines.

Ten leukemic cell line were used for determination of drug efficacy: KASUMI1 (ref. ACC 220) , MONOMAC6 (ref. ACC 124), KASUMI3 (ref. ACC 714), OCI-AML3 (ref. ACC 582), MOLM14 (ref. ACC 777), THP1 (ref. ACC 16) HL60 (ref. ACC 3), K562 (ref. ACC 10) and KG1 (ref. ACC 14) were purchased from Leibniz Institute DSMZ (Braunschweig, Germany). MV4.11 cell line was generously given by Institute of Molecular Genetics of the Czech Academy of Sciences (IMG)- Prague. All cell lines were cultured using the medium and supplements as recommended by the suppliers. The cell lines were maintained at 37°C in an incubator with humidified air with 5% CO2.

Determination of Drugs’ inhibitory growth Concentration 50% (IG50) using Cell Viability assays.

Cells were seeded (10,000-20,000 cells/well) into 96-well cell culture plates at concentrations required to ensure approximately 80% confluence in control (untreated cells). Cells were grown 24 hours using their corresponding culture medium as recommended by manufacturers and were maintained at 37°C in an incubator with humidified air with 5% CO2. Then, DIMATE, its structural analogues and other standard of care drugs were added with increasing concentration. DIMATE was used from 0,05pM to 20pM; Decitabine from 0,05pM to 1000pM; Azacitidine from 0,05pM to 500pM; S55746 from 0,25pM to 5pM; Hydrea from 0,5pM to 2500pM; All Trans Retinoic Acid (ATRA) from 0,01 pM to 100pM; Arsenic TriOxide (ATO) from 0,05pM to 100pM; Daunorubicine from 0,5pM to 400pM; Cytarabine from 0,05pM to 200pM; Pracinostat from 0,5pM to 100pM; Venetoclax from 0,16pM to 2,5pM The growth-inhibitory effect of the drugs was analyzed using an in vitro, resazurin-based toxicity assay (ab129732, Abeam, Cambridge UK). 30pl of 1X resazurine, diluted in PBS, was distributed per well. Cells were incubated for 3 hours at 37°C and absorbance was read at 570nm. The drug response was quantified by the half- maximal inhibitory growth concentration (IG50) and determined by non-linear regression analysis of log-dose/response curves using GraphPad Prism software (GraphPad software, San Diego California USA).

Drug-drug interaction assays.

Initially, single compound dose-response curves were performed for DIMATE, DIMATE’s structural analogues, Decitabine, Azacitidine, S55746, Hydrea, ATRA (All Trans Retinoic Acid), ATO (Arsenic TriOxide), Daunorubicine, Cytarabine, Praci nostat and Venetoclax and the corresponding the IG50 was calculated for each cancer cell line. Subsequently, for each compound (i.e. Decitabine, Azacitidine, S55746, Hydrea, ATRA, ATO, Daunorubicine, Cytarabine, Pracinostat and Venetoclax) six concentrations, including the concentration corresponding to their respective IG50, were selected to be combined with six different concentrations of DIMATE or its structural analogues. In the case of HMA (i.e. Decitabine and Azacitidine), the combination was carried out sequentially. That is, AML cells were firstly pre-treated overnight (18h) with HMA alone at different concentrations (0.05/1/5/10/30500pM for Azacitidine; 0.05/1/10/50/250/1 OOOpM for Decitabine). After this incubation, DIMATE (0.05/1/3/5/10/20pM) or DIMATE structural analogues were added to the cells together with HMA so that each of the 6 concentration of HMA was tested against the 6 concentrations of DI MATE (6x6 matrix). Cells were incubated for an additional 24h. Drugs used for the synergy study were the fallowing one: aracytine (#C3350000), Daunorubicine (#D0125000), Azacytidine (#A2385), Azacytidine (#A2385), Arsenic Trioxide (ATO, #202673) and All Trans Retinoic acid (ATRA, #R2625), all were purchased from Sigma Aldrich-Merck. Decitabine (#AB-M2052) and Pracinostat (#AB-M1778) were purchased from AbMole. Hydrea has been generously given by Hospital de la Conception, Marseille. Venetoclax (ABT-199, AB-M20175-5mg) and S55746 (SE-S8759-5mg) were purchased from Euromedex. DIMATE was from Advanced BioDesign : Azacytidine (#A2385) was purchased from Sigma Aldrich-Merck; and Decitabine (#AB-M2052) was purchased from AbMole.

Synergistic interactions were assessed using the Loewe additivity model to derive Loewe Synergy scores (LS); with a LS > 10 indicating synergy. Synergism was assessed by calculating the proportion of cell growth using SynergyFinder 2.0: visual analytics of multidrug combination synergies. All pics of synergy (the highest value of Loewe score) were retained to create a dendrogram using R software (https://www.r-project.org/) (....). The dendogram ranked AML cell lines according to their corresponding LS value.

Hematopoietic colony culture assays.

Colony culture assays were performed using Methocult GF M3434 (Stemcell Technologies, Vancouver, BC, Canada). AML cells obtained from bone marrow aspirates of refractory/relapse AML patients samples were plated in petri dishes using syringe and blunt- end needle at the following quantities: 3000 cells for patient#1 , 3000 cells for patient#2 and 20000 cells for patient#3, following Methocult manufacturer’s instructions. Cells were subsequently exposed to DIMATE (1uM and 100nM) alone and/or in combination with the HMA agents 5-Azacitidine (250nM) or decitabine (10nM) (MedChem Tronica, Sollentuna, Sweden). Cells were incubated in a humidified incubator at 370C and 5%CO2. The resulting colonies were counted using a bright-field microscope, after 14 days of in vitro culture. Counting was performed by investigators blinded to the treatment.

Results The synergistic effect of combinations according to the invention is clearly demonstrated by Figures 1 ,2 and 3.

Claims

1 A compound of formula (I): wherein:

R-, and R₂, identical or different, are chosen among a C C₁₀ alkyl group, a phenyl, a benzyl, CHR₅CHR₆OR₄ and (CHR₅)_VOR_4I or RT and R₂ together with the nitrogen atom to which they are linked form an heterocycle, in particular a piperidine or a morpholine; said phenyl and benzyl being optionally substituted by one or more substituents chosen from: linear or branched (C₁-C₇)alkyl, halogen, NO₂ and CONH₂ ;

R₃ is chosen from linear or branched (C₁-C₇)alkyl,

R₅ and R₆ identical or different are independently chosen from:

• H and linear or branched (CrC₇)alkyl, or

- _v is chosen from 2 to 4;

R₇ is -(C₁-C₃)alkyl;

R₈ is -(C₁-C₃)alkylNRR’; R and R’ identical or different, are independently chosen from H and linear or branched (Ci-C₇)alkyl, or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers; in combination with an Hypomethylating Agent (HMA), for use for the prevention and/or treatment of cancer in a patient.

2.- Products comprising a compound of formula (I): wherein:

R₃ is chosen from linear or branched (C C₇)alkyl,

R₅ and R₆ identical or different are independently chosen from:

• H and linear or branched (Ci -C₇)alkyl , or

• R₆ is H and R-, and R₅ are linked together to R-, to form with the nitrogen atom linked to R-, an heterocycloalkyl; - _v is chosen from 2 to 4;

R₇ is -(Ci-C₃)alkyl;

R₈ is -(Ci-C₃)alkylNRR’;

R and R’ identical or different, are independently chosen from H and linear or branched (C₁-C₇)alkyl, or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers; and an Hypomethylating Agent (HMA) as a combined preparation for use spread out over time for use for the prevention and/or treatment of cancer in a patient.

3.- A pharmaceutical composition comprising a compound of formula (I): wherein:

R₃ is chosen from linear or branched (C C₇)alkyl,

R₅ and R₆ identical or different are independently chosen from:

• H and linear or branched (CrC₇)alkyl, or

- _v is chosen from 2 to 4;

R₇ is -(C₁-C₃)alkyl;

R₈ is -(C₁-C₃)alkylNRR’;

4.- A compound of formula (I) and an HMA for use according to claim 1 , products for use according to claim 2 or pharmaceutical composition according to claim 3, wherein the compound of formula (I) is characterized in that R₃ is linear or branched (CrC₇)alkyl, preferably methyl, R-, is linear or branched (C C₇)alkyl, preferably methyl, R₂ is a C C₁₀ alkyl group, preferably a methyl, CHR₅CHR₆OR₄, (CHR₅)_VOR₄ or R-i and R₂ together with the nitrogen atom to which they are linked form an heterocycle, in particular a morpholine, and R₅ and _R6 are H.

5.- A compound of formula (I) and an HMA for use according to claim 1 or 4, products for use according to claim 2 or 4, or pharmaceutical composition according to claim 3 or 4, wherein the compound of formula (I) is characterized in that R₅ and R₆ are H and R₄ is chosen from (C₂-C₇) cycloalkyl, linear or branched -(C₁-C₇)alkyl-heteroaryl, or benzyl; preferably benzyl; said (C₂-C₇) cycloalkyl being substituted by one or more substituents chosen from: linear or branched (C C₇)alkyl, said benzyl being optionally substituted by one or more substituents chosen from: linear or branched (C C₇)alkyl optionally substituted by one or more halogen atom, linear or branched (C C₇)alkoxy optionally substituted by one or more halogen atom, halogen.

6.- A compound of formula (I) and an HMA for use according to any one of claims 1 and 4 to 5, products for use according to any one of claims 2 and 4 to 5, or pharmaceutical composition according to any one of claims 3 to 5, wherein the compound of formula (I) is chosen from:

S-methyl 4-[2-ethoxyethyl(methyl)amino]-4-methyl-pent-2-ynethioate; S-methyl 4-[2-allyloxyethyl(methyl)amino]-4-methyl-pent-2-ynethioate;

S-methyl 4-[2-benzyloxyethyl(methyl)amino]-4-methyl-pent-2-ynethioate;

S-methyl 4-methyl-4-[methyl-[2-(m-tolylmethoxy)ethyl]amino]pent-2-ynethioate;

S-methyl 4-[2-[(3,4-dimethylphenyl)methoxy]ethyl-methyl-amino]-4-methyl-pent-2- ynethioate;

S-methyl 4-[2-[(4-methoxyphenyl)methoxy]ethyl-methyl-amino]-4-methyl-pent-2- ynethioate;

S-methyl 4-[2-[(3,4-dimethoxyphenyl)methoxy]ethyl-methyl-amino]-4-methyl-pent- 2-ynethioate;

S-methyl 4-[2-[(3-fluorophenyl)methoxy]ethyl-methyl-amino]-4-methyl-pent-2- ynethioate;

S-methyl 4-methyl-4-[methyl-[2-(2-pyridylmethoxy)ethyl]amino]pent-2-ynethioate;

S-methyl 4-methyl-4-[methyl-[2-(3-pyridylmethoxy)ethyl]amino]pent-2-ynethioate;

S-methyl 4-methyl-4-[methyl-[2-(4-pyridylmethoxy)ethyl]amino]pent-2-ynethioate;

S-methyl 4-((4-(benzyloxy)butyl)(methyl)amino)-4-methylpent-2-ynethioate;

S-methyl 4-((2-hydroxyethyl)(methyl)amino)-4-methylpent-2-ynethioate;

S-methyl 4-methyl-4-[methyl-[2-(2-naphthylmethoxy)ethyl]amino]pent-2-ynethioate;

S-methyl 4-methyl-4-[methyl-[2-[(2,6,6-trimethylcyclohexen-1- yl)methoxy]ethyl]amino]pent-2-ynethioate;

S-methyl 2,5,10,11 ,11 -pentamethyl-6-oxo-7-oxa-2,5, 10-triazatetradec-12-yne- 14- th ioate;

S-methyl 4-methyl-4-[methyl(2-phenoxycyclopentyl)amino]pent-2-ynethioate;

(S)-S-methyl 4-(2-((benzyloxy)methyl)pyrrolidin-1-yl)-4-methylpent-2-ynethioate;

S-methyl 4-[3(benzyloxy)-1 pyrrolidinyl])-4-methylpent-2-ynethioate;

S-methyl 1- 4-Dimethylamino-4-methyl-pent-2-ynethioate;

S-methyl 5- 4-Methyl-4-morpholin-4-yl-pent-2-ynethioate;

S methyl 4-methyl-4- [methyl (octyl) amino] pent-2-ynethioate; or its pharmaceutically acceptable salts or optical isomers, racemates, diastereoisomers, enantiomers or tautomers.

7.- A compound of formula (I) and an HMA for use according to any one of claims 1 and 4 to 6, products for use according to any one of claims 2 and 4 to 6, or pharmaceutical composition according to any one of claims 3 to 6, wherein the compound of formula (I) is comprised in a lipidic nanocapsule.

8.- A compound of formula (I) and an HMA for use according to claim 7, products for use according to claim 7, or pharmaceutical composition according to claim 7, wherein said lipidic nanocapsule comprises:

- an oily core comprising between 25 and 90% by weight of medium chain triglycerides, preferably between 60 and 80%, relative to the total weight of the nanocapsule, and the compound of formula (I); and

- a shell surrounding the oily core, comprising between 3 and 25% by weight relative to the total weight of the nanocapsule of at least one lipid surfactant, and at least one hydrophilic surfactant; and

• in which the ratio by weight relative to the total weight of the nanocapsule between the medium chain triglycerides and the compound of formula (I) is of at least 4.

9.- A compound of formula (I) and an HMA for use according to any one of claims 1 and 4 to 8, products for use according to any one of claims 2 and 4 to 8, or pharmaceutical composition according to any one of claims 3 to 8, wherein said HMA is chosen from Azacitidine, Decitabine, Guadecitabine, CC-486, decitabine, guadecitabine, SGI-110, CP- 4200, NTX-301 or ASTX727.

10.- A compound of formula (I) and an HMA for use according to any one of claims 1 and 4 to 9, or products for use according to any one of claims 2 and 4 to 9, wherein said cancer is a is a relapsed or refractory cancer.

11 .- A compound of formula (I) and an HMA for use according to any one of claims 1 and 4 to 10, or products for use according to any one of claims 2 and 4 to 10, wherein the patient has been previously treated with a chemotherapy-based regimen, in particular a daunorubicin based regimen.

12.- A compound of formula (I) and an HMA for use according to any one of claims 1 and 4 to 11 , or products for use according to any one of claims 2 and 4 to 11 , wherein said cancer is chosen from hematological and solid cancers.

13. - A compound of formula (I) and an HMA for use according to any one of claims 1 and 4 to 12, or products for use according to any one of claims 2 and 4 to 12, wherein said cancer is chosen from Acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), Myeloproliferative Neoplasm (MPN), Lymphoma non-Hodgkin and T-cell, Large B-Cell Diffuse Lymphoma, Lung cancer, Colorectal cancer, including microsatellite stable colorectal cancer, Breast Cancer, including patients with advanced or metastatic HER2- negative breast cancer, and patients with triple-negative breast cancer, Liver and hepatic Cancer, Prostate cancer, Epithelial Ovarian Cancer, Pancreatic and esophageal/gastric adenocarcinoma (EGC), Skin Neoplasm, Kidney/Renal Cancer, Urothelial or Bladder Carcinoma Anorexia Nervosa, Testis Cancer, and Glioblastoma.

14. - A compound of formula (I) and an HMA for use according to any one of claims 1 and 4 to 13, or products for use according to any one of claims 2 and 4 to 13, wherein the cancer is AML.

15. - A compound of formula (I) and an HMA for use according to claim 14, or products for use according to claim 14, wherein the patient is also treated with a Bcl2 protein family inhibitor, in particular venetoclax.