NOVEL PHARMACOLOGICAL CHAPERONE COMPOUNDS OF HUMAN ACID ALPHA-GLUCOSIDASE AND THE THERAPEUTIC USE THEREOF
20230339856 · 2023-10-26
Inventors
- Sandrine PY (SAINT MARTIN D'URIAGE, FR)
- Alice KANAZAWA (BERNIN, FR)
- Anaïs VIEIRA DA CRUZ (MOIRANS, FR)
- Salia TANGARA (LILLE, FR)
Cpc classification
C07D405/12
CHEMISTRY; METALLURGY
C07D401/04
CHEMISTRY; METALLURGY
C07D211/46
CHEMISTRY; METALLURGY
C07D455/02
CHEMISTRY; METALLURGY
International classification
C07D211/46
CHEMISTRY; METALLURGY
C07D401/04
CHEMISTRY; METALLURGY
C07D401/12
CHEMISTRY; METALLURGY
C07D405/12
CHEMISTRY; METALLURGY
C07D455/02
CHEMISTRY; METALLURGY
Abstract
The invention concerns particular compounds of the iminosugars class, with a piperidine ring, having a D-gluco configuration and comprising a quaternary centre in the a position of the nitrogen of the piperidine ring. These compounds have the ability to stabilize human acid α-glucosidase while being selective with respect to other glycosidases. They are particularly advantageous for use as chaperone molecules of this enzyme for the treatment of Pompe disease.
Claims
1. A method of treating a subject, comprising administering to said subject a therapeutically-effective amount of a compound of general formula (I), or one of the pharmaceutically acceptable salts thereof: ##STR00031## wherein R.sup.1 represents a hydrogen atom or a linear, branched and/or cyclic hydrocarbon radical, saturated or unsaturated, aromatic or not, optionally substituted, optionally comprising one or more heteroatoms and/or one or more groups including at least one heteroatom and optionally comprising a single ring or a plurality of rings optionally fused, and R.sup.2 represents a linear, branched and/or cyclic hydrocarbon radical, saturated or unsaturated, aromatic or not, optionally substituted, optionally comprising one or more heteroatoms and/or one or more groups including at least one heteroatom and optionally comprising a single ring or a plurality of rings optionally fused, said hydrocarbon radical comprising at least 2 carbon atoms when R.sup.1 represents a hydrogen atom, or R.sup.1 and R.sup.2 form together, with the atoms of the piperidine ring to which each is attached, a 3-to 6-membered heterocycle fused with the piperidine ring, optionally substituted by one or several radicals, which may be identical or different, each selected from the group consisting of a hydroxyl group, an amino group or a linear, branched and/or cyclic carbon radical, saturated or unsaturated, aromatic or not, optionally substituted, optionally comprising one or more heteroatoms and/or one or more groups including at least one heteroatom and optionally comprising a single ring or a plurality of rings optionally fused.
2. The method of claim 1, wherein, in the general formula (I): R.sup.1 represents a hydrogen atom or a linear, branched and/or cyclic hydrocarbon radical, saturated or unsaturated, aromatic or not, optionally substituted, optionally comprising one or more heteroatoms and/or one or more groups including at least one heteroatom and optionally comprising a single ring or a plurality of rings optionally fused, and R.sup.2 represents a —CH(R.sup.3)—R.sup.4 group, wherein R.sup.3 and R.sup.4, identical or different, each represent a hydrogen atom or a linear, branched and/or cyclic hydrocarbon radical, saturated or unsaturated, aromatic or not, optionally substituted, optionally comprising one or more heteroatoms and/or one or more groups including at least one heteroatom and optionally comprising a single ring or a plurality of rings optionally fused, R.sup.3 and R.sup.4 not simultaneously representing a hydrogen atom when R.sup.1 represents a hydrogen atom.
3. The method of claim 1, wherein, in the general formula (I): R.sup.1 represents a hydrogen atom or a linear, branched and/or cyclic hydrocarbon radical, saturated or unsaturated, aromatic or not, optionally substituted, optionally comprising one or more heteroatoms and/or one or more groups including at least one heteroatom and optionally comprising a single ring or a plurality of rings optionally fused, and R.sup.2 represents a —(CHR.sup.7)—SO.sub.2—Ar.sup.1 group wherein Ar.sup.1 represents an aryl or heteroaryl radical, optionally substituted, and R.sup.7 represents a hydrogen atom or a linear, branched and/or cyclic hydrocarbon radical, saturated or unsaturated, aromatic or not, optionally substituted, optionally comprising one or more heteroatoms and/or one or more groups including at least one heteroatom and optionally comprising a single ring or a plurality of rings optionally fused.
4. The method of claim 1, wherein, in the general formula (I): R.sup.1 represents a hydrogen atom or a linear, branched and/or cyclic hydrocarbon radical, saturated or unsaturated, aromatic or not, optionally substituted, optionally comprising one or more heteroatoms and/or one or more groups including at least one heteroatom and optionally comprising a single ring or a plurality of rings optionally fused, and R.sup.2 represents a triazole group, optionally substituted.
5. The method of claim 1, wherein, in the formula (I): R.sup.1 represents a hydrogen atom or a linear, branched and/or cyclic hydrocarbon radical, saturated or unsaturated, aromatic or not, optionally substituted, optionally comprising one or more heteroatoms and/or one or more groups including at least one heteroatom and optionally comprising a single ring or a plurality of rings optionally fused, and R.sup.2 represents a —(CH.sub.2).sub.2—R.sup.8 group, wherein R.sup.8 represents: a hydrogen atom; a C1-C12 alkyl or cycloalkyl group, optionally substituted, optionally comprising a single ring or a plurality of fused rings; a —(CH.sub.2).sub.a—OH group wherein a is an integer between 0 and 18; a —(CH.sub.2).sub.b—Ar.sup.2 group wherein Are represents an aryl or heteroaryl radical, optionally substituted, optionally comprising one or more heteroatoms and/or one or more groups including at least one heteroatom and optionally comprising a single ring or a plurality of rings optionally fused and b is an integer between 0 and 18; a —(CH.sub.2).sub.c—Si(R.sup.9).sub.3 group wherein R.sup.9 represents a hydroxyl radical, a C1-C4 alkyl radical, a C1-C4 alkoxyl radical or a phenyl radical and c is an integer between 0 and 18; or a —(CH.sub.2).sub.d13 Z—R.sup.10 group wherein Z is a heteroatom selected from the group consisting of oxygen, nitrogen and sulfur, R.sup.10 represents a hydrogen atom or a C1-C18 alkyl, cycloalkyl, alkylaryl, aryl or acyl radical, said radical optionally being interrupted and/or substituted by one or more heteroatoms and/or one or more groups including at least one heteroatom, and d is an integer between 0 and 18.
6. The method of claim 1, wherein, in the general formula (I), R.sup.1 represents a C1-C18, linear, branched and/or cyclic alkyl group, optionally interrupted and/or substituted by one or more heteroatoms and one or more groups including at least one heteroatom.
7. The method of claim 1, wherein said compound has the general formula (I′a): ##STR00032## wherein: R.sup.1 represents a hydrogen atom or C1-C18 linear, branched and/or cyclic alkyl group, optionally interrupted and/or substituted by one or more heteroatoms and/or one or more groups including at least one heteroatom, R.sup.8 represents: a hydrogen atom; a methyl, ethyl, propyl, butyl, pentyl, hexyl, cycloalkyl, adamantyl, alkylcycloalkyl, alkylaryl or aryl radical, said radical optionally being interrupted and/or substituted by one or more heteroatoms and/or one or more groups including at least one heteroatom; a hydroxyl group; a —Si(R.sup.12).sub.3 group wherein R.sup.12 represents a hydroxyl radical, a C1-C4 alkyl radical, a C1-C4 alkoxyl radical or a phenyl radical; a —(CH.sub.2).sub.f—Y—R.sup.13 group wherein f is an integer between 0 and 6, Y is a heteroatom selected from the group consisting of oxygen, nitrogen and sulfur and R.sup.13 represents a C1-C18 alkyl radical or a C1-C18 aryl or heteroaryl radical; a —(CH.sub.2).sub.g—CO—R.sup.13 group wherein g is an integer between 0 and 6; or a —(CH.sub.2).sub.h—SO.sub.e—R.sup.13 group wherein h is an integer between 0 and 6 and e is equal to 1 or 2, R.sup.8 not representing a hydrogen atom when R.sup.1 represents a propyl radical.
8. The method of claim 1, of wherein said compound has the general formula (I″a): ##STR00033## wherein: R.sup.1 represents a hydrogen atom or a C1-C18 linear, branched and/or cyclic alkyl group, optionally interrupted and/or substituted by one or more heteroatoms and/or one or more groups including at least one heteroatom, R.sup.18 represents a linear, branched and/or cyclic hydrocarbon radical, saturated or unsaturated, aromatic or not, optionally substituted, comprising 4 to 18 carbon atoms, optionally comprising one or more heteroatoms and/or one or more groups including at least one heteroatom and optionally comprising a single ring or a plurality of rings optionally fused.
9. The method of claim 1, wherein said compound has the general formula (I′b): ##STR00034## wherein R.sup.14 represents a hydrogen atom, a carbonyl radical or a C1-C18 alkyl, alkenyl, alkynyl, alkylaryl or aryl radical, said radical optionally being interrupted and/or substituted by one or more heteroatoms and/or one or more groups including at least one heteroatom, and R.sup.15 represents a hydrogen atom, a hydroxyl radical, an amino radical, or a C1-C18 alkyl, alkenyl or aryl radical, optionally interrupted and/or substituted by one or more heteroatoms and/or one or more groups including at least one heteroatom.
10. The method of claim 1, wherein, in the general formula (I), R.sup.1 and R.sup.2 form together, with the atoms of the piperidine ring to which each is attached, a 3-membered heterocycle fused with the piperidine ring, optionally substituted by a —X—R.sup.5 group, wherein: X represents a —C(═O)— or —CH(OR.sup.6)— radical wherein R.sup.6 represents a linear, branched and/or cyclic hydrocarbon radical, saturated or unsaturated, aromatic or not, optionally substituted, optionally comprising one or more heteroatoms and/or one or more groups including at least one heteroatom and optionally comprising a single ring or a plurality of rings optionally fused, and R.sup.5 represents a hydrogen atom, an amino group, or a linear, branched and/or cyclic hydrocarbon radical, saturated or unsaturated, aromatic or not, optionally substituted, optionally comprising one or more heteroatoms and/or one or more groups including at least one heteroatom and optionally comprising a single ring or a plurality of rings optionally fused, or X represents a —CH(OH)— radical and R.sup.5 represents a hydrogen atom or a linear, branched and/or cyclic hydrocarbon radical, saturated or unsaturated, aromatic or not, optionally substituted, optionally comprising one or more heteroatoms and/or one or more groups including at least one heteroatom and optionally comprising a single ring or a plurality of rings optionally fused.
11. The method of claim 1, which is for treating Pompe disease in said subject.
12. The method as claimed in claim 11, which is for stabilizing human acid α-glucosidase in said subject.
13. (canceled)
14. A pharmaceutical composition containing a compound of general formula (I) or one of the pharmaceutically acceptable salts thereof in a pharmaceutically acceptable vehicle; ##STR00035## wherein R.sup.1 represents a hydrogen atom or a linear, branched and/or cyclic hydrocarbon radical, saturated or unsaturated, aromatic or not, optionally substituted, optionally comprising one or more heteroatoms and/or one or more groups including at least one heteroatom and optionally comprising a single ring or a plurality of rings optionally fused, and R.sup.2 represents a linear, branched and/or cyclic hydrocarbon radical, saturated or unsaturated, aromatic or not, optionally substituted, optionally comprising one or more heteroatoms and/or one or more groups including at least one heteroatom and optionally comprising a single ring or a plurality of rings optionally fused, said hydrocarbon radical comprising at least 2 carbon atoms when R.sup.1 represents a hydrogen atom, or R.sup.1 and R.sup.2 form together, with the atoms of the piperidine ring to which each is attached, a 3-to 6-membered heterocycle fused with the piperidine ring, optionally substituted by one or several radicals, which may be identical or different, each selected from the group consisting of a hydroxyl group, an amino group or a linear, branched and/or cyclic carbon radical, saturated or unsaturated, aromatic or not, optionally substituted, optionally comprising one or more heteroatoms and/or one or more groups including at least one heteroatom and optionally comprising a single ring or a plurality of rings optionally fused.
15. The pharmaceutical composition of claim 14, which is in a form suitable for oral administration.
16. A compound of general formula (I′): ##STR00036## wherein R.sup.1 represents a hydrogen atom or a linear, branched and/or cyclic hydrocarbon radical, saturated or unsaturated, aromatic or not, optionally substituted, optionally comprising one or more heteroatoms and/or one or more groups including at least one heteroatom and optionally comprising a single ring or a plurality of rings optionally fused, and R.sup.2 represents a —CH(R.sup.3)—R.sup.4 group, wherein R.sup.3 and R.sup.4, identical or different, each represent a hydrogen atom or a linear, branched and/or cyclic hydrocarbon radical, saturated or unsaturated, aromatic or not, optionally substituted, optionally comprising one or more heteroatoms and/or one or more groups including at least one heteroatom and optionally comprising a single ring or a plurality of rings optionally fused, R.sup.3 and R.sup.4 being such that, when R.sup.1 and R.sup.3 each represent a hydrogen atom, R.sup.4 does not represent a hydrogen atom or a phenyl radical, and R.sup.1 and R.sup.2 being such that they do not simultaneously represent, respectively, a propyl radical and an ethyl radical, or R.sup.1 and R.sup.2 form together, with the atoms of the piperidine ring to which each is attached, a 4-membered heterocycle fused with the piperidine ring, optionally substituted by one or more radicals, which may be identical or different, each selected from the group consisting of a hydroxyl group, an amino group or a linear, branched and/or cyclic carbon radical, saturated or unsaturated, aromatic or not, optionally substituted, optionally comprising one or more heteroatoms and/or one or more groups including at least one heteroatom and optionally comprising a single ring or a plurality of rings optionally fused, or R.sup.1 and R.sup.2 form together, with the atoms of the piperidine ring to which each is attached, a 3-membered heterocycle fused with the piperidine ring, optionally substituted by a —X—R.sup.5 group, wherein: X represents a —C(═O)— or —CH(OR.sup.6)— radical wherein R.sup.6 represents a linear, branched and/or cyclic hydrocarbon radical, saturated or unsaturated, aromatic or not, optionally substituted, optionally comprising one or more heteroatoms and/or one or more groups including at least one heteroatom and optionally comprising a single ring or a plurality of rings optionally fused, and R.sup.5 represents a hydrogen atom, an amino group or a linear, branched and/or cyclic hydrocarbon radical, saturated or unsaturated, aromatic or not, optionally substituted, optionally comprising one or more heteroatoms and/or one or more groups including at least one heteroatom and optionally comprising a single ring or a plurality of rings optionally fused; or X represents a —CH(OH)— radical and R.sup.5 represents a hydrogen atom or a linear, branched and/or cyclic hydrocarbon radical, saturated or unsaturated, aromatic or not, optionally substituted, optionally comprising one or more heteroatoms and/or one or more groups including at least one heteroatom and optionally comprising a single ring or a plurality of rings optionally fused, R.sup.5 not representing an n-propyl radical, a cyclohexyl radical or a phenyl radical, or one of the pharmaceutically acceptable salts thereof.
17. The compound of claim 16, wherein, in the formula (I′): R.sup.1 represents a hydrogen atom or a linear, branched and/or cyclic hydrocarbon radical, saturated or unsaturated, aromatic or not, optionally substituted, optionally comprising one or more heteroatoms and/or one or more groups including at least one heteroatom and optionally comprising a single ring or a plurality of rings optionally fused, and R.sup.2 represents a —(CHR.sup.7)—SO.sub.2—Ar.sup.1 group wherein Ar.sup.1 represents an aryl or heteroaryl radical, optionally substituted, and R.sup.7 represents a hydrogen atom or a linear, branched and/or cyclic hydrocarbon radical, saturated or unsaturated, aromatic or not, optionally substituted, optionally comprising one or more heteroatoms and/or one or more groups including at least one heteroatom and optionally comprising a single ring or a plurality of rings optionally fused.
18. 1 The compound of claim 16, wherein, in the formula (I′): R.sup.1 represents a hydrogen atom or a linear, branched and/or cyclic hydrocarbon radical, saturated or unsaturated, aromatic or not, optionally substituted, optionally comprising one or more heteroatoms and/or one or more groups including at least one heteroatom and optionally comprising a single ring or a plurality of rings optionally fused, and R.sup.2 represents a triazole group, optionally substituted.
19. The compound of claim 16, wherein, in the formula (I′): R.sup.1 represents a hydrogen atom or a linear, branched and/or cyclic hydrocarbon radical, saturated or unsaturated, aromatic or not, optionally substituted, optionally comprising one or more heteroatoms and/or one or more groups including at least one heteroatom and optionally comprising a single ring or a plurality of rings optionally fused, and R.sup.2 represents a —(CH.sup.2).sub.2—R.sup.8 group, wherein R.sup.8 represents: a hydrogen atom; a C1-C12 alkyl or cycloalkyl group, optionally substituted, optionally comprising a single ring or a plurality of fused rings; a —(CH.sub.2).sub.a—OH group wherein a is an integer between 0 and 18; a —(CH.sub.2).sub.b—Ar.sup.2 group wherein Are represents an aryl or heteroaryl radical, optionally substituted, optionally comprising one or more heteroatoms and/or one or more groups including at least one heteroatom and optionally comprising a single ring or a plurality of rings optionally fused and b is an integer between 0 and 18; a —(CH.sub.2).sub.c—Si(R.sup.9).sub.3 group wherein R.sup.9 represents a hydroxyl radical, a C1-C4 alkyl radical, a C1-C4 alkoxyl radical or a phenyl radical and c is an integer between 0 and 18, or a —(CH.sub.2).sub.d—Z—R.sup.1° group wherein Z is a heteroatom selected from the group consisting of oxygen, nitrogen and sulfur, R.sup.10 represents a C1-C18 alkyl, cycloalkyl, alkylaryl, aryl or acyl radical, said radical optionally being interrupted and/or substituted by one or more heteroatoms and/or one or more groups including at least one heteroatom, and d is an integer between 0 and 18.
20. The compound of claim 16, of general formula (I′a): ##STR00037## wherein R.sup.1 represents a hydrogen atom or a C1-C18 linear, branched and/or cyclic alkyl group, optionally interrupted and/or substituted by one or more heteroatoms and/or one or more groups including at least one heteroatom, R.sup.8 represents: a hydrogen atom; a hydroxyl group; a methyl, ethyl, propyl, butyl, pentyl, hexyl, cyclohexyl, adamantyl, alkylcycloalkyl, alkylaryl or aryl radical, said radical optionally being interrupted and/or substituted by one or more heteroatoms and/or one or more groups including at least one heteroatom; a —Si(R.sup.12).sub.3 group wherein R.sup.12 represents a hydroxyl radical, a C1-C4 alkyl radical, a C1-C4 alkoxyl radical or a phenyl radical; a —(CH.sub.2).sub.f—Y—R.sup.13 group wherein f is an integer between 0 and 6, Y is a heteroatom selected from the group consisting of oxygen, nitrogen and sulfur and R.sup.13 represents a C1-C18 alkyl radical or a C1-C18 aryl or heteroaryl radical; a —(CH.sub.2).sub.g—CO—R.sup.13 group wherein g is an integer between 0 and 6; or a —(CH.sub.2).sub.h—SO.sub.e—R.sup.13 group wherein h is an integer between 0 and 6 and e is equal to 1 or 2, R.sup.8 not representing a hydrogen atom when R.sup.1 represents a propyl radical.
21. The compound of claim 16, of general formula (I″a): ##STR00038## wherein: R.sup.1 represents a hydrogen atom or a C1-C18 linear, branched and/or cyclic alkyl group, optionally interrupted and/or substituted by one or more heteroatoms and/or one or more groups including at least one heteroatom, R.sup.18 represents a linear, branched and/or cyclic hydrocarbon radical, saturated or unsaturated, aromatic or not, optionally substituted, comprising from 4 to 18 carbon atoms, optionally comprising one or more heteroatoms and/or one or more groups including at least one heteroatom and optionally comprising a single ring or a plurality of rings optionally fused.
22. The compound of claim 16, of formula (I′b): ##STR00039## wherein R.sup.14 represents a hydrogen atom, a carbonyl radical or a C1-C18 alkyl, alkenyl, alkynyl, alkylaryl or aryl radical, said radical optionally being interrupted and/or substituted by one or more heteroatoms and/or one or more groups including at least one heteroatom, and R.sup.15 represents a hydrogen atom, a hydroxyl radical, an amino radical or a C1-C18 alkyl, alkenyl or aryl radical, optionally interrupted and/or substituted by one or more heteroatoms and/or one or more groups including at least one heteroatom.
23. The compound of claim 16, of formula (I′c): ##STR00040## wherein R.sup.16 represents a hydrogen atom or a C1-C18 alkyl radical, said radical optionally being interrupted and/or substituted by one or more heteroatoms and/or one or more groups including at least one heteroatom, R.sup.16 not representing an n-propyl radical, a cyclohexyl radical or a phenyl radical.
24. A method for preparing the compound of claim 19, comprising successive steps of: a/ reacting a compound of general formula (III) or (IV) of formulae: ##STR00041## wherein Bn represents a benzyl radical, with a compound of general formula (V): ##STR00042## wherein R.sup.8 is as defined in claim 19, R.sup.8 however representing neither a hydrogen atom, nor a hydroxyl group, nor an amino group, in the presence of an organometallic compound, b/ optionally, reduction of the hydroxylamine function into amine, c/ optionally, alkylation of the nitrogen atom of the piperidine ring, d/ and hydrogenolysis of the product obtained at the end of step a/, where applicable of step b/ or of step c/.
Description
BRIEF DESCRIPTION OF THE DRAWINGS
[0226] The features and advantages of the invention will emerge more clearly in the light of the following example embodiments, provided by simple way of illustration and in no way !imitative of the invention, with the support of
[0227]
[0228]
[0229]
[0230]
[0231]
[0232]
[0233]
[0234]
DESCRIPTION OF THE PREFERRED EMBODIMENTS
A/ Synthesis and Characterization of Compounds Used in Accordance With the Invention
[0235] All the reactions were conducted under inert atmosphere in previously dried glassware, under stirring by a magnetic bar, the assembly being placed above a magnetic stirrer, except in the case of reactions under irradiation by microwave. The latter were implemented in sealed tubes equipped with a magnetic bar placed in a microwave reactor, at a temperature regulated by an internal infrared probe. The toluene, the ether and the dichloromethane used as reaction solvents were previously filtered on an Inert PureSolv® purification system. The acetonitrile and dichloroethane were distilled on CaH.sub.2. The THF was distilled on sodium in the presence of benzophenone. The commercial reagents, the methanol and the ethanol were used without purification. The reactions were followed by thin-layer chromatography (TLC) by means of silica on aluminum plates (Merck, Kieselgel 60 F254), with revelation under ultraviolet radiation and then by a 3% solution of potassium permanganate in a 10% solution of potassium hydroxide (w/v). The purifications by column chromatography were implemented by means of Macherey-Nagel® Silica Gel 60 (70-230 mesh). The rotatory powers were measured on a PerkinElmer 341 polarimeter. The infrared spectra were obtained on a Nicolet “Magna 550” spectrometer including an ATR (attenuated total reflexion) module on which the products were deposited pure, in the solid or liquid state. The data are expressed in cm.sup.−1. The .sup.1H NMR and .sup.13C NMR {DEPT-Q} were obtained with Avance 500 (.sup.1H: 500 MHz, .sup.13C: 125 MHz) or Avance 400 (1H: 400 MHz, .sup.13C: 100 MHz) spectrometers. The chemical shifts for the .sup.1H spectra are expressed with respect to those of the residual solvents contained in CDCl.sub.3 (δ7.26 ppm) or CD.sub.3OD (δ 3.31 ppm). The chemical shifts for the .sup.13C spectra are expressed with respect to those of the solvents CDCl.sub.3 (δ 77.16 ppm) or CD.sub.3OD (δ 49.00 ppm). The .sup.1H NMR spectra are reported as follows: chemical shift (ppm), multiplicity (br: broad; s: singulet; d: doublet; dd: doublet of doublets; t: triplet; pst: pseudo triplet; m: multiplet), coupling constants (Hz) and integration. The high-resolution mass spectra (HRMS) were recorded on a Thermo Scientific ESI/LTQ Orbitrap XL® or Waters G2-S Q-TOF mass spectrometer.
A.1/Synthesis Methods 1
[0236] The synthesis methods used for obtaining compounds complying with the above general formula (I′a) falls under the general synthesis schemes shown on
1) General Protocol A—Alkynylation of the Nitrone of Formula (III) Leading to the Propargylic Hydroxylamines of Formula A-1
[0237] A 1M solution of diethyl zinc in hexane (1.5 equiv.) is added dropwise to an alkyne solution of formula (V) (4 equiv.) in anhydrous toluene at 0° C. under argon atmosphere. The resulting mixture is stirred for 30 minutes at 0° C. A solution of ketonitrone of formula (III) (1 equiv.) in anhydrous toluene is then added dropwise at 0° C., and then the reaction mixture is stirred until the reaction is complete (followed by TLC). A saturated aqueous solution of NaHCO.sub.3 is added and then the resulting mixture is diluted with diethylether. The organic phase is separated and the aqueous phase is extracted twice with diethylether. The organic phases are washed with brine, dried on MgSO.sub.4 and evaporated under reduced pressure. The hydroxylamines of formula A-1 thus formed are purified by silica gel chromatography, before being reduced and debenzylated in accordance with the following general protocol B.
2) General Protocol A′—Alkynylation of the Nitrone of Formula (III) Leading to the Propargylic Hydroxylamines of Formula A-1.
[0238] A 1.4 M solution of n-butyllithium in hexane (2.5 equiv.) is added dropwise to a solution of alkyne of formula (V) (2.5 equiv.) in anhydrous THF at −10° C. under argon atmosphere. The resulting solution is stirred for 15 minutes at −10° C. and is then cooled to −78° C. A solution of ketonitrone of formula (III) (1 equiv.) in anhydrous THF is then added dropwise at −78° C., and then the reaction mixture is stirred until the reaction is complete (followed by TLC). A saturated aqueous solution of NH.sub.4Cl is added and the mixture is allowed to rise to ambient temperature, and then extracted three times by ethyl acetate. The organic phases are collected and washed with brine, dried on MgSO.sub.4 and concentrated under reduced pressure. The hydroxylamines of formula A-1 thus isolated are purified by silica gel chromatography, before being reduced and debenzylated in accordance with the following general protocol B.
3) General Protocol B
[0239] 1.5 equiv. of hydrochloric acid HCI (2M solution in anhydrous ether) and 0.2 equiv. of Pd/C 10% are added to a solution of O-benzylated iminosugar (1 equiv.) in absolute ethanol. The suspension obtained is stirred at room temperature for 17 hours under pressure of hydrogen (5 bar) and then filtered on Celite®. The Celite® is rinsed with methanol and the filtrate is concentrated at reduced pressure to give an iminosugar hydrochlorate. This salt is purified on Dowex® 50W-X2 ion exchange resin previously activated by HCI, and eluted with aqueous NH.sub.4OH, to obtain after evaporation of the solvents the corresponding neutral compound of formula (I′a).
Compound of Formula (IIc)
[0240] The compound of above formula (IIc) ((2R,3R,4R,5S)-2-(hydroxymethyl)-2-phenethylpiperidine-3,4,5-triol: 36.7 mg; 73% for 2 steps) was prepared in accordance with the general procedures A and B, from nitrone of formula (III) (110.0 mg, 0.205 mmol) and phenylacetylene (Va: R.sup.8=Ph, 0.034 mL; 0.307 mmol).
[0241] A solid yellow is obtained, with the following characteristics:
[0242] [α].sup.20.sub.D+4.7 (c 0.98, CH.sub.3OH);
[0243] IR v 3291, 2918, 2864, 1632, 1494, 1455, 1363, 1270, 1198, 1094, 1076, 1040, 1004 cm.sup.−1;
[0244] .sup.1H NMR (500 MHz, CD.sub.3OD) δ 7.31-7.08 (m, 5H), 3.73 (d, J=10.4 Hz, 1H), 3.55 (d, J=10.5 Hz, 1H), 3.55-3.43 (m, 2H), 3.45-3.34 (m, 1H), 3.00-2.90 (m, 1H), 2.68-2.51 (m, 3H), 1.91-1.72 (m, 2H) ppm;
[0245] .sup.13C NMR (125 MHz, CD.sub.3OD) δ 144.4 (.sup.ArCq), 129.4 .sup.(ArCH), 129.3 (.sup.ArCH), 126.7(.sup.ArCH), 76.5 (CH), 75.2 (CH), 73.8 (CH), 65.6 (CH.sub.2), 60.5 (Cq), 45.9 (CH.sub.2), 30.7 (CH.sub.2), 29.9 (CH.sub.2) ppm;
[0246] HRMS (ESI.sup.+) Calc. C.sub.14H.sub.22NO.sub.4 [M+H].sup.+: m/z=268.15433; Found m/z=268.15372.
Compound (IIx1)
[0247] The compound of above formula (IIx1) ((2R,3R,4R,5S)-2-(hydroxymethyl)-2-(2-(trimethylsilyl)ethyl)piperidine-3,4,5-triol: 7.1 mg; 29% for 2 steps) was prepared in accordance with the general procedures A and B, from nitrone of formula (III) (102.1 mg, 0.189 mmol) and trimethylsilylacetylene (Vb: R.sup.8=TMS, 0.105 mL; 0.759 mmol).
[0248] A white foam is obtained, with the following characteristics:
[0249] [hd].sup.20D+4.51 (c 0.71, MeOH);
[0250] .sup.1H NMR (400 MHz, CD.sub.3OD) δ 3.57 (d, J=10.9 Hz, 1H), 3.52 (d, J=9.3 Hz, 1H), 3.44 (t, J=4.0 Hz, 1H), 3.39 (d, J=10.0 Hz, 1H), 3.39-3.33 (m, 1H), 2.85 (dd, J=12.8, 5.5 Hz, 1H), 2.42 (dd, J=12.8, 10.8 Hz, 1H), 1.62-1.51 (m, 2H), 0.56-0.44 (m, 1H), 0.39-0.29 (m, 1H), 0.02 (s, 9H) ppm;
[0251] .sup.13C NMR (100 MHz, CD.sub.3OD) δ 76.6 (CH), 75.5 (CH), 73.8 (CH), 65.6 (CH.sub.2), 60.5 (Cq), 45.8 (CH.sub.2), 21.0 (CH.sub.2), 8.60 (CH.sub.2), −1.86 (CH.sub.3) ppm;
[0252] HRMS (ESI.sup.+) Calc. C.sub.11H.sub.26NO.sub.4Si [M+H].sup.+: m/z=264.16256; Found m/z=264.16282.
Compound (IIr)
[0253] The compound of above formula (IIr) ((2R,3R,4R,5S)-2-(2-((3R,5R,7R)-adamantan-1-ypethyl)-2-(hydroxymethyl)piperidine-3,4,5-triol: 15.0 mg; 27% for 2 steps) was prepared in accordance with the general procedures A′ and B, from nitrone of formula (III) (100 mg, 0.185 mmol) and adamantylacetylene (Vh: R.sup.8=adamantyl, 74.5 mg, 0.465 mmol).
[0254] A yellowish lacquer is obtained, with the following characteristics:
[0255] [α].sup.20.sub.D+7.46 (c 0.63, MeOH);
[0256] .sup.1H NMR (500 MHz, CD.sub.3OD) δ 3.79 (d, J=10.9 Hz, 1H), 3.64-3.48 (m, 4H), 3.11 (br d, J=11.3 Hz, 1H), 2.78-2.66 (m, 1H), 2.00-1.91 (m, 3H), 1.82-1.63 (m, 8H), 1.58-1.49 (m, 6H), 1.19-1.03 (m, 2H) ppm;
[0257] .sup.13C NMR (100 MHz, CD.sub.3OD) δ 74.9 (CH), 72.5 (CH), 70.8 (CH), 63.7 (Cq), 62.5 (CH.sub.2), 44.2 (CH.sub.2), 43.3 (CH.sub.2), 38.2 (CH.sub.2), 37.0 (CH.sub.2), 30.1 (CH), 20.7 (CH.sub.2) ppm;
[0258] HRMS (ESI.sup.+) Calc. C.sub.18H.sub.31NO.sub.4 [M+H].sup.+: m/z=326.23258; Found m/z=326.23225.
Compound (IIb7)
[0259] The compound of above formula (IIb7) ((2R,3R,4R,5S)-2-(2-cyclohexylethyl)-2-(hydroxymethyl)piperidine-3,4,5-triol: 57.3 mg; 59% for 2 steps) was prepared in accordance with the general procedures A and B, from nitrone of formula (III) (99.8 mg; 0.185 mmol) and 1-ethynylcyclohexene (Vd: R.sup.8=cyclohexene; 87 μL; 0.742 mmol).
[0260] A white solid is obtained, with the following characteristics:
[0261] [α].sup.20.sub.D+3.63 (c 1.43, MeOH);
[0262] .sup.1H NMR (400 MHz, CD.sub.3OD) δ 3.60 (d, J=10.6 Hz, 1H, .sup.7CH.sub.2), 3.52-3.43 (m, 2H, .sup.3CH, .sup.4CH), 3.43-3.33 (m, 2H), 2.87 (dd, J=12.8, 5.0 Hz), 2.49 (ps t, J=11.7 Hz, 1H), 1.83-1.62 (m, 5H), 1.60-1.47 (m, 2H), 1.34-1.06 (m, 6H), 1.01-0.86 (m, 2H) ppm;
[0263] .sup.13C NMR (100 MHz, CD.sub.3OD) δ 76.5 (CH), 75.1 (CH), 73.8 (CH), 65.5 (CH.sub.2), 60.3 (Cq), 45.8 (CH.sub.2), 39.9 (CH), 34.6 (CH.sub.2), 34.5 (CH.sub.2), 30.6 (CH.sub.2), 27.8 (CH.sub.2), 27.5 (CH.sub.2), 24.8 (CH.sub.2) ppm;
[0264] HRMS (ESI.sup.+) Calc. C.sub.14H.sub.20NO.sub.4 [M+H].sup.+: m/z=274.20128; Found m/z=274.20139.
Compound (IIb1)
[0265] The compound of above formula (IIb1) ((2R,3R,4R,5S)-2-(hydroxymethyl)-2-pentylpiperidine-3,4,5-triol: 32.8 mg; 67% for 2 steps) was prepared in accordance with the general procedures A and B, from nitrone of formula (III) 96.8 mg; 0.180 mmol) and 1-pentyne (Ve: R.sup.8=C.sub.3H.sub.7; 71 μL; 0.720 mmol).
[0266] A yellow solid is obtained, with the following characteristics:
[0267] [α].sup.20.sub.D−1.29 (c 1.01, MeOH);
[0268] .sup.1H NMR (400 MHz, CD.sub.3OD) δ 3.61 (d, J=10.6 Hz, 1H), 3.52-3.33 (m, 4H), 2.87 (dd, J=12.8, 5.3 Hz, 1H), 2.49 (ps t, J=11.8 Hz, 1H), 1.62-1.43 (m, 2H), 1.43-1.17 (m, 6H), 0.92 (t, J=6.7 Hz, 3H) ppm;
[0269] .sup.13C NMR (100 MHz, CD.sub.3OD) δ 76.5 (CH), 75.2 (CH), 73.9 (CH), 65.7 (CH.sub.2), 60.2 (Cq), 45.9 (CH.sub.2), 33.9 (CH.sub.2), 27.7 (CH.sub.2), 23.6 (CH.sub.2), 22.9 (CH.sub.2), 14.4 (CH.sub.3) ppm; HRMS (ESI.sup.+) Calc. C.sub.11H.sub.24NO.sub.4 [M+H]+: m/z=234.16998; Found m/z=234.17004.
Compound (IIb3)
[0270] The compound of above formula (IIb3) ((2R,3R,4R,5S)-2-heptyl-2-(hydroxymethyl)piperidine-3,4,5-triol: 33.2 mg; 69% for 2 steps) was prepared in accordance with the general procedures A and B, from nitrone of formula (III) (100.7 mg; 0.187 mmol) and 1-heptyne (Vf: R.sup.8=C.sub.5H.sub.11; 98 μL; 0.749 mmol).
[0271] A yellow solid is obtained, with the following characteristics:
[0272] [α].sup.20.sub.D+1.04 (c 0.96, MeOH);
[0273] 1H NMR (400 MHz, CD.sub.3OD) δ 3.61 (d, J=10.6 Hz, 1H), 3.51-3.44 (m, 2H), 3.41 (d, J=10.8 Hz), 3.39-3.33 (m, 1H), 2.87 (dd, J=13.0, 5.4 Hz, 1H), 2.49 (ps t, J=11.4 Hz, 1H), 1.58-1.47 (m, 2H), 1.43-1.18 (m, 10H), 0.90 (t, J=6.2 Hz, 3H) ppm;
[0274] .sup.13C NMR (100 MHz, CD.sub.3OD) δ 76.5 (CH), 75.1 (CH), 73.8 (CH), 65.6 (CH.sub.2), 60.3 (Cq), 45.9 (CH.sub.2), 33.0 (CH.sub.2), 31.7 (CH.sub.2), 30.4 (CH.sub.2), 23.7 (CH.sub.2), 23.2 (CH.sub.2), 27.8 (CH.sub.2), 14.4 (CH.sub.3) ppm;
[0275] HRMS (ESI.sup.+) Calc. C.sub.13H.sub.28NO.sub.4 [M+H].sup.+: m/z=261.20128; Found m/z=261.20131.
Compound (IIc1)
[0276] The compound of above formula (IIc1) ((2R,3R,4R,5S)-2-(hydroxymethyl)-2-(3-phenylpropyl)piperidine-3,4,5-triol: 20.2 mg; 45% for 2 steps) was prepared according to the general procedures A and B, from nitrone of formula (III) (96.7 mg; 0.179 mmol) and 3-phenyl-1-propyne (Vg: R.sup.8=CH.sub.2Ph; 89 μL; 0.719 mmol). A yellow lacquer is obtained, with the following characteristics:
[0277] [α].sup.20.sub.D+1.68 (c 1.37, MeOH);
[0278] .sup.1H NMR (400 MHz, CD.sub.3OD) δ 7.10-7.10 (m, 5H), 3.60 (d, J=10.6 Hz, 1H), 3.49-3.37 (m, 3H), 3.37-3.32 (m, 1H), 2.84 (dd, J=13.1, 5.4 Hz, 1H), 2.69-2.53 (m, 2H), 2.42 (dd, J=13.0, 10.8 Hz, 1 H), 1.75-1.49 (m, 4H) ppm;
[0279] .sup.13C NMR (100 MHz, CD.sub.3OD) δ 143.6 (.sup.ArCq), 129.4 (.sup.ArCH), 129.3 (.sup.ArCH), 126.7 (.sup.ArCH), 76.5 (CH), 75.1 (CH), 73.8 (CH), 65.6 (CH.sub.2), 60.3 (Cq), 45.8 (CH.sub.2), 37.6 (CH.sub.2), 27.5 (CH.sub.2), 25.3 (CH.sub.2) ppm;
[0280] HRMS (ESI.sup.+) Calc. C.sub.15H.sub.24NO.sub.4 [M+H]+: m/z=282.16998; Found m/z=282.16968.
A.2/ Synthesis Method 2
[0281] The synthesis method used for obtaining compounds complying with the general formula (I′a) below falls under the general synthesis scheme 2 shown on
Compound of Formula (IId)
[0282] The compound of above formula (IId) (2R,3R,4R,5S)-2-(2-hydroxyethyl)-2-(hydroxymethyl)piperidine-3,4,5-triol is prepared in the following manner.
[0283] A mixture of nitrone (III) (566 mg, 1.05 mmol) and of trimethylsilylacetylene alkyne (Vb: R.sup.8=SiMe.sub.3, 2.2 mL, 15.75 mmol) was stirred at ambient temperature for 21 hours, and then the excess alkyne was evaporated under reduced pressure. The raw isoxazoline B-2 thus obtained (667 mg, 1.05 mmol) was dissolved in anhydrous THF (20 mL) and a solution of TBAF (1M in THF, 1.05 mL, 1.05 mmol) was added at 0° C. The solution was stirred at 0° C. for 45 minutes, and then CH.sub.2Cl.sub.2 and water were added. The aqueous phase was extracted three times with CH.sub.2Cl.sub.2. The organic phases were washed with brine, dried on MgSO.sub.4, and then the solvents were evaporated under reduced pressure. Purification of the residue obtained by chromatography supplied bicyclic β-lactic C-2 (440.6 mg, 74% in 2 steps). LiAlH.sub.4 (9.6 mg, 0.25 mmol) was added at 0° C. to a solution of this β-lactam C-2 (71.3 mg, 0.13 mmol) in ether (1.5 mL). The mixture was stirred at ambient temperature for 2 hours and then diluted with CH.sub.2Cl.sub.2, aqueous NH.sub.4Cl and a few drops of aqueous NaOH up to basic pH. The aqueous phase was extracted (three times) with CH.sub.2Cl.sub.2. The organic phases were washed with brine, dried on MgSO.sub.4, and the solvents were evaporated under reduced pressure to give piperidine D-2 (translucent oil, 61.1 mg, 85%) after purification by chromatography. This compound (26.3 mg, 0.463 mmol) was debenzylated in accordance with the general procedure B to give piperidine (IId) (9.3 mg, 97%). A pale yellow lacquer was obtained, with the following characteristics:
[0284] [α].sup.20.sub.D+8.2 (c 0.61, CH.sub.3OH);
[0285] IR v 3287, 2920, 1644, 1431, 1081 cm.sup.−1;
[0286] .sup.1H NMR (500 MHz, CD.sub.3OD) δ 1.86 (t, J=6.5 Hz, 2H), 2.65 (dd, J=11.2, 12.6 Hz, 1H), 2.92 (dd, J=5.4, 13.1 Hz, 1H), 3.33-3.43 (m, 2H), 3.48 (d, J=9.1 Hz, 1H), 3.58 (d, J=10.8 Hz, 1H), 3.64-3.76 (m, 3H) ppm;
[0287] .sup.13C NMR (125 MHz, CD.sub.3OD) δ 30.5 (CH.sub.2), 45.8 (CH.sub.2), 58.8 (CH.sub.2), 60.9 (C.sub.q), 65.7 (CH.sub.2), 73.5 (CH), 74.6 (CH), 76.3 (CH) ppm;
[0288] HRMS (ESI.sup.+) Calc. C.sub.8H.sub.18NO.sub.5[M+H].sup.+: m/z=208.1185; Found m/z=208.1184.
A.3/ Synthesis Method 3
[0289] The synthesis method used for obtaining compounds complying with the general formula (I′d) below falls under the general synthesis scheme 3 shown on
Compound of Formula (IIw)
[0290] The compound of above formula (IIw) (2S,3S,4R,5S)-3,4,5-tris(benzyloxy)-2-((benzyloxy)methyl)-2-(pyridin-2-ylsulfonyl)piperidin-1-ol is prepared in the following manner. A solution of nitrone of formula (III) (196 mg; 0.366 mmol) and of 2-pyridyl-methylsulfone (Vla) (74.6 mg; 0.475 mmol) in anhydrous THF (10 mL) under argon atmosphere was cooled to −78° C. 1M LiHMDS in THF (548 μL; 0.548 mmol) was added dropwise and then the reaction mixture was stirred at this same temperature until the reaction was complete. After the addition of water and ethyl acetate, the aqueous phase was extracted (three times) with ethyl acetate. The collected organic phases were washed with brine, dried on MgSO.sub.4 and then concentrated at reduced pressure. Purification of the residue obtained by chromatography supplied the compound of formula A-3a (R.sup.7=H) 115 mg; 55%). This compound was debenzylated according to a variant of the general procedure B to give piperidine (IIw).
A.4/ Synthesis Methods 4
[0291] The synthesis methods used for obtaining compounds complying with the general formula (I), in particular compounds complying with the general formula (I′a), below fall under a general synthesis scheme 4 shown on
Compound of Formula (IIa)
[0292] The compound of above formula (IIa) ((2R,3R,4R,5S)-2-ethyl-2-(hydroxymethyl)piperidine-3,4,5-triol) was prepared in accordance with the general protocol B described for the synthesis method 1, from piperidine D-4b (R.sup.17=vinyl, 74.0 mg, 0.137 mmol), obtained as described in the publication by Boisson et al., 2015, Org. Lett. 17: 3662-3665, and was isolated with a yield of 95% (24.9 mg).
[0293] A pale yellow lacquer is obtained, with the following characteristics:
[0294] [α].sup.20.sub.D+1.6 (c 1.22, CH.sub.3OH);
[0295] IR v 3286, 2939, 1643, 1445, 1096 cm.sup.−1;
[0296] .sup.1H NMR (500 MHz, CD.sub.3OD) δ 0.86 (t, J=7.6 Hz, 3H), 1.55-1.68 (m, 2H), 2.51 (dd, J=10.8, 12.8 Hz, 1H), 2.89 (dd, J=5.4, 13.0 Hz, 1H), 3.34-3.40 (m, 1H), 3.42 (d, J=10.7 Hz, 1H), 3.44-3.51 (m, 2H), 3.62 (d, J=10.7 Hz, 1H) ppm; .sup.13C NMR (125 MHz, CD.sub.3OD) δ 7.0 (CH.sub.3), 20.1 (CH.sub.2), 45.7 (CH.sub.2), 60.5 (C.sub.q), 65.0 (CH.sub.2), 73.6 (CH), 75.1 (CH), 76.4 (CH) ppm;
[0297] HRMS (ESI.sup.+) Calc. C.sub.8H.sub.17NO.sub.4[M+H].sup.+: m/z=192.1236; Found m/z=192.1237.
Compound of Formula (IIb)
[0298] The compound of above formula (IIb) ((2R,3R,4R,5S)-2-(hydroxymethyl)-2-propylpiperidine-3,4,5-triol) was prepared in accordance with the general protocol B described for the synthesis method 1, from piperidine D-4a (R.sup.17=allyl, 39.7 mg, 0.071 mmol), obtained as described in the publication by Vieira Da Cruz et al., 2017, J. Org. Chem. 82: 9866-9872, and was isolated with a yield of 79% (11.4 mg).
[0299] A colorless lacquer is obtained, with the following characteristics:
[0300] [α].sup.20.sub.D+5.4 (c 0.55, CH.sub.3OH);
[0301] IR v 3291, 2932, 1625, 1454, 1090 cm.sup.−1;
[0302] .sup.1NMR (500 MHz, CD.sub.3OD) δ 0.94 (t, J=7.2 Hz, 3H), 1.22-1.42 (m, 2H), 1.49-1.57 (m, 2H), 2.53 (dd, J=10.8, 13.0 Hz, 1H), 2.89 (dd, J=5.5, 13.0 Hz, 1H), 3.34-3.41 (m, 1H), 3.42 (d, J=10.7 Hz, 1H), 3.45-3.51 (m, 2H), 3.64 (d, J=10.7 Hz, 1H) ppm;
[0303] .sup.13C NMR (125 MHz, CD.sub.3OD) δ 15.3 (CH.sub.3), 16.6 (CH.sub.2), 30.4 (CH.sub.2), 45.7 (CH.sub.2), 60.7 (CO, 65.3 (CH.sub.2), 73.5 (CH), 74.9 (CH), 76.3 (CH) ppm;
[0304] HRMS (ESI.sup.+) Calc. C.sub.9H.sub.20NO.sub.4 [M+H].sup.+: m/z=206.1392; Found m/z=206.1396
Compound of Formula (IIe)
[0305] The compound of above formula (IIe) (2R,3R,4R,5S)-2-ethyl-2-(hydroxymethyl)-1-propylpiperidine-3,4,5-triol is prepared in the following manner.
[0306] The piperidine E-4b (R.sup.1=allyl, R.sup.17=vinyl; 62 mg, 0.105 mmol), obtained as described in the publication by Boisson et al., 2015, Org. Lett. 17: 3662-3665, was dissolved in anhydrous acetonitrile (0.5 mL), treated with 2-nitrobenzenesulfonyl chloride (186 mg, 0.84 mmol) at 0° C., and then with hydrazine monohydrate (0.08 mL, 1.68 mmol), added dropwise at 0° C. This reaction mixture was maintained under stirring at ambient temperature for 28 hours, and then water was added and the aqueous phase was extracted (3 times) with dichloromethane. The collected organic phases were washed with brine, dried over MgSO.sub.4 and concentrated under reduced pressure. After purification by chromatography, a mixture of the products of hydrogenation solely of the allyl group and of hydrogenation of the allyl and vinyl groups was isolated (58 mg, 94%, 3:7 mixture). This mixture was dissolved in methanol (1.5 mL), and then treated with Pearlman's reagent (20% Pd(OH).sub.2/C, 16.8 mg, 0.119 mmol) and HCl (2 M solution in ether, 0.2 mL, 0.4 mmol) under hydrogen atmosphere (1 atm) and under vigorous stirring for 40 hours. The mixture was filtered over Celite®, the Celite® was rinsed several times with methanol, then the filtrate was concentrated under reduced pressure. The residue was solubilized in water and purified on cation exchange resin (DOWEX 50W-X8, form H.sup.+; elution by an aqueous solution of 1M NH.sub.4OH) to provide the product (IIe) (11.9 mg, 49%). A beige solid is obtained, with the following characteristics:
[0307] [α].sup.20.sub.D−40.7 (c 0.60, CH.sub.3OH);
[0308] IR v 3363, 2962, 2932, 2874, 2826, 1653, 1464, 1379, 1097, 1048, 1016 cm.sup.−1 .sup.1H NMR (500 MHz, CD.sub.3OD) δ□0.90 (t, J=7.5 Hz, 3H), 0.96 (t, J=7.5 Hz, 3H), 1.39-1.50 (m, 1H), 1.51-1.60 (m, 1H), 1.61-1.75 (m, 2H), 2.26-2.34 (m, 1H), 2.42-2.51 (m, 1H), 2.76-2.85 (m, 1H), 2.93-3.00 (m, 1H), 3.40-3.52 (m, 3H), 3,65 (d, J=11.0 Hz, 1H), 3.79 (d, J=11.0 Hz, 1H) ppm;
[0309] .sup.13C NMR (125 MHz, CD.sub.3OD) δ 9.6 (CH.sub.3), 11.9 (CH.sub.3), 19.2 (CH.sub.2), 23.2 (CH.sub.2), 52.0 (CH.sub.2), 52.2 (CH.sub.2), 62.5 (CH.sub.2), 71.5 (CH), 74.9 (CH), 76.4 (CH) ppm;
[0310] HRMS (ESI.sup.+) Calc. C.sub.11H.sub.24NO.sub.4 [M+H].sup.+: m/z=234.1700; Found m/z=234.1696.
Compound of Formula (IIf)
[0311] The compound of formula (IIf) as above (1R,2R,3S,9aR)-9a-(hydroxymethyl)octahydro-1H-quinolizine-1,2,3-triol is prepared in the following manner.
[0312] Piperidine E-4a (R.sup.1=R.sup.17=allyl; 35 mg, 0.06 mmol), obtained as described in the publication by Vieira Da Cruz et al., 2017, J. Org. Chem. 82: 9866-9872, was dissolved in anhydrous CH.sub.2Cl.sub.2 (3 mL). This solution was carefully degassed, and then the Grubbs II catalyst (1.3 mg, 2.5 mol %) was added. The mixture was maintained under stirring at ambient temperature for 20 hours, and then filtered over silica. The silica was rinsed with ethyl acetate and methanol, and the solvents were evaporated under reduced pressure. After purification of the residue thus obtained by chromatography, the product of cyclization by metathesis of F-4a olefins (32 mg, 95%) was isolated. The latter (34 mg, 0.06 mmol) was dissolved in methanol (1 mL), and then treated with Pearlman's reagent (20% Pd(OH).sub.2/C, 14 mg, 0.10 mmol) and HCl (2M solution in ether, 0.045 mL, 0.09 mmol) under hydrogen atmosphere (5 bar) and under vigorous stirring for 17 hours. The mixture was filtered over Celite®, the Celite®was rinsed several times with methanol, and then the filtrate was concentrated under reduced pressure. The residue was dissolved in water and purified over cation exchange resin (DOWEX® 50W-X8, form H.sup.+; elution by an aqueous solution of 1M NH.sub.4OH) to provide the product (IIf) (9 mg, 73%).
[0313] A colorless oil is obtained, with the following characteristics:
[0314] [α].sup.20.sub.D+16.6 (c 0,41, CH.sub.3OH);
[0315] IR v 3336, 2942, 2869, 1053 cm.sup.−1;
[0316] .sup.1H NMR (500 MHz, CD.sub.3OD) δ 1.28 (br d, J=11.6 Hz, 1H), 1.37-1.44 (m, 1H), 1.62-1.76 (m, 3H), 1.81-1.94 (m, 1H), 2.66 (dd, J=3.0, 14.0 Hz, 1H), 2.72 (dd, J=4.9, 11.3 Hz, 1H), 3.12 (t, J=11.0 Hz, 1H), 3.20 (pstd, J=3.0, 14.0 Hz, 1H), 3.38 (pst, J=9.3 Hz, 1H), 3.45-3.52 (m, 2H), 3.68 (d, J=11.1 Hz, 1H), 3.95 (d, J=11.1 Hz, 1H) ppm;
[0317] .sup.13C NMR (125 MHz, CD.sub.3OD) δ 18.7 (CH.sub.2), 19.2 (CH.sub.2), 20.7 (CH.sub.2), 48.5 (CH.sub.2), 53.3 (CH.sub.2), 60.7 (CH.sub.2), 62.0 (C.sub.q), .sub.71.4 (CH), 74.1 (CH), 75.9 (CH) ppm; HRMS (ESI.sup.+) Calc. C.sub.10H.sub.20NO.sub.4 [M+H].sup.+: m/z=218.1392; Found m/z=218.1396.
Compound of Formula (IIg)
[0318] The compound of formula (IIg) as above (6S, 7R, 8R, 8aR)-8a-(hydroxymethyl)-octahydroindolizine-6,7,8-triol is prepared in the following manner.
[0319] Piperidine E-4b (R.sup.1=allyl, R.sup.17=vinyl; 74 mg, 0.125 mmol), obtained as described in the publication by Boisson et al., 2015, Org. Lett. 17: 3662-3665, was dissolved in anhydrous CH.sub.2Cl.sub.2 (6.3 mL). This solution was carefully degassed, and then the Grubbs II catalyst (5.3 mg, 0.006 mmol) was added and the mixture was heated at 40° C. for 3 hours, under stirring. After cooling to ambient temperature, the reaction mixture was filtered over silica, the silica was rinsed with ether, and the solvents were evaporated under reduced pressure. After purification of the residue thus obtained by chromatography, the product of cyclization by metathesis of olefins F-4b (65 mg, 93%) was isolated. The latter (50 mg, 0.089 mmol) was dissolved in methanol (1.4 mL), then treated with Pearlman's reagent (20% Pd(OH).sub.2/C, 17.9 mg, 0.025 mmol) and HCl (2M solution in ether, 0.066 mL, 0.133 mmol) under hydrogen atmosphere (5 bar) and under vigorous stirring for 17 h. The mixture was filtered over Celite®, the Celite® was rinsed several times with methanol, and then the filtrate was concentrated under reduced pressure. The residue was dissolved in water and purified over cation exchange resin (DOWEX 50W-X8, form H.sup.+; elution by an aqueous solution of 1M NH.sub.4OH) to provide the product (IIg) (16 mg, 89%).
[0320] A beige solid is obtained, with the following characteristics:
[0321] [α].sup.20.sub.D+27.9 (c 0.41, CH.sub.3OH);
[0322] IR v 3330, 2924, 1652, 1031 cm.sup.−1;
[0323] .sup.1H NMR (500 MHz, CD.sub.3OD) δ 1.46-1.53 (m, 1H), 1.67-1.76 (m, 1H), 1.88-1.96 (m, 1H), 1.98-2.08 (m, 1H), 2.46 (pst, J=11.0 Hz, 1H), 2.80-2.89 (m, 2H), 3.09-3.18 (m, 1H), 3.33-3.41 (m, 2H), 3.43-3.49 (m, 1H), 3.53 (d, J=11.5 Hz, 1H), 3.76 (d, J=9.5 Hz, 1H) ppm;
[0324] .sup.13C NMR (125 MHz, CD.sub.3OD) δ 21.0 (CH.sub.2), 26.1 (CH.sub.2), 53.8 (CH.sub.2), 55.9 (CH.sub.2), 63.6 (CH.sub.2), 71.0 (CH), 73.0 (CH), 73.2 (Cq), 76.5 (CH) ppm;
[0325] HRMS (ESI.sup.+) Calc. C.sub.9H.sub.18NO.sub.4 [M+H].sup.+: m/z=204.1236; Found m/z=204.1234.
A.5/ Synthesis Methods 5
[0326] The synthesis method used for obtaining the compound complying with the general formula (I′b) as above falls under a general synthesis scheme 5 shown on
Compound (IIh)
[0327] The compound of above formula (IIh) ((3S,4R,5R,6R)-6-(hydroxymethyl)-1-azabicyclo[4.2.0]octane-3,4,5-triol) is prepared in the following manner.
[0328] [Rh(COD).sub.2]BF.sub.4 (2.5 mg, 0.006 mmol), 1,3-bis(diphenylphosphino)propane (2.7 mg, 0.006 mmol) and PhSiH.sub.3 (25 μL, 0.20 mmol) were added to a solution of β-lactam C-5 (corresponding to the compound C-2 obtained as intermediate in synthesis method 2, as described in the protocol for preparing the compound (IId) above) (57.1 mg, 0.10 mmol) in distilled THF. The solution was stirred at 50° C. for 4 hours. After cooling, the reaction mixture was diluted with EtOAc and aqueous NaOH (1 M). The aqueous phase was extracted (three times) with EtOAc. The organic phases were washed with brine, dried over MgSO.sub.4, and the solvents were evaporated under reduced pressure to give conidine G-5 (33.2 mg, 60%) after purification by chromatography. This compound (20.0 mg, 0.04 mmol) was debenzylated in accordance with the general procedure B to give the compound (IIh) (6.3 mg, 93%).
[0329] A pale yellow oil is obtained, with the following characteristics:
[0330] [α].sup.20.sub.D−5.17 (c 0.58, CH.sub.3OH);
[0331] IR v 3234, 2917, 1429, 1030 cm.sup.−1;
[0332] .sup.1H NMR (500 MHz, CD.sub.3OD) δ 2.18-2.26 (m, 1H), 2.55-2.63 (m, 1H), 2.89 (dd, J=5.9, 13.3 Hz, 1H), 3.21 (dd, J=5.1, 13.3 Hz, 1H), 3.62 (d, J=11.7 Hz, 1H), 3.69-3.80 (m, 5H), 3.99 (pst, J=6.5 Hz, 1H) ppm;
[0333] .sup.13C NMR (125 MHz, CD.sub.3OD) δ 21.3 (CH.sub.2), 51.4 (CH.sub.2), 52.9 (CH.sub.2), 65.5 (CH.sub.2), 71.2 (CH), 72.7 (CH), 73.2 (C.sub.q), 76.8 (CH) ppm;
[0334] HRMS (ESI.sup.+) Calc. C.sub.8H.sub.16NO.sub.4 [M+H].sup.+m/z=190.1074; Found m/z=190.1074.
[0335] General Protocol C—reducing alkylation of β-lactam C-5 leading to conidines of general formula (I′b)
[0336] A solution of β-lactam C-5 (1 equiv.) and of Vaska's catalyst (4% mol) in anhydrous dichloromethane under argon atmosphere is added to tetramethyldisiloxane (TMDS, 2 equiv.) at ambient temperature. The mixture is stirred for 45 minutes and then cooled to −78° C. Grignard reagent (2 equiv.) is added dropwise and then the mixture is stirred for 7-10 minutes at this same temperature before increasing to ambient temperature. It is next stirred for 21 hours. A saturated aqueous solution of NH.sub.4Cl is added and then the resulting mixture is diluted with CH.sub.2Cl.sub.2. The organic phase is separated and the aqueous phase is extracted twice with CH.sub.2Cl.sub.2. The organic phases are washed with brine, dried over MgSO.sub.4 and evaporated under reduced pressure. The conidines of formula (I′b) thus formed are purified by chromatography over silica gel before being reduced and debenzylated in accordance with the general protocol B above.
Compound (IIz2)
[0337] The compound of above formula (1Iz2) ((3S,4R,5R,6R,8R)-8-benzyl-6-(hydroxymethyl)-1-azabicyclo[4.2.0]octane-3,4,5-triol: 20.3 mg; 65% for 2 steps) was prepared in accordance with the general procedures C and B, from β-lactam C-5 (70.1 mg; 0.124 mmol) and 2M benzylmagnesium chloride in THF (R.sup.14=Bn; 36.3 μL; 0.248 mmol).
[0338] A translucent lacquer is obtained, with the following characteristics:
[0339] .sup.1H NMR (500 MHz, CD.sub.3OD) δ 7.39-7.20 (m, 5H), 4.45-4.34 (m, 1H), 3.95 (t, J=5.7 Hz, 1H), 3.80-3.74 (m, 2H), 3.70 (d, J=5.6 Hz, 1H), 3.56 (d, J=12.1 Hz, 1H), 3.21-3.12 (m, 2H), 3.00 (dd, J=13.6, 8.4 Hz, 1H), 2.94 (dd, J=13.1, 6.5 Hz, 1H), 2.60 (dd, J=11.9, 9.0 Hz, 1H), 2.32-2.20 (m, 1H) ppm;
[0340] .sup.13C NMR (125 MHz, CD.sub.3OD) δ 137.1 (.sup.ArCq), 130.1 (.sup.ArCH), 129.7 (.sup.ArCH), 128.0 (.sup.ArCH), 76.5 (CH), 71.5 (CH), 71.1 (CH), 65.7 (CH), 64.1 (CH.sub.2), 51.4 (CH.sub.2), 41.0 (CH.sub.2), 27.3 (CH.sub.2) ppm;
[0341] HRMS (ESI.sup.+) Calc. C15H22NO.sub.4 [M+H].sup.+: m/z=280.15433; Found m/z=280.15437.
Compound (IIz3)
[0342] The compound of above formula (IIz3) ((3S,4R,5R,6R,8R)-6-(hydroxymethyl)-8-methyl-1-azabicyclo[4.2.0]octane-3,4,5-triol: 8.5 mg; 46% for 2 steps) was prepared in accordance with the general procedures C and B, from β-lactam C-5 (192.6 mg; 0.342 mmol) and 3M methylmagnesium chloride in THF (R.sup.14=Me; 228 μL; 0.683 mmol).
[0343] A yellowish solid is obtained, with the following characteristics:
[0344] [60 ].sup.20.sub.D−22.9 (c 1.07, MeOH);
[0345] .sup.1H NMR (500 MHz, CD.sub.3OD) δ 3.99 (t, J=6.7 Hz, 1H), 3.85-3.75 (m, 1H), 3.75-3.69 (m, 1H), 3.67 (d, J=7.6 Hz, 1H), 3.54 (d, J=11.2 Hz, 1H), 3.47 (d, J=11.2 Hz, 1H), 3.10 (dd, J=13.9, 5.5 Hz, 1H), 2.71 (dd, J=13.8, 4.0 Hz, 1H), 2.58 (dd, J=11.5, 8.9 Hz, 1H), 1.64 (dd, J=10.9, 8.0 Hz, 1H), 1.21 (d, J=6.1 Hz, 3H) ppm;
[0346] .sup.13C NMR (100 MHz, CD.sub.3OD) δ 77.2 (CH), 74.7 (CH), 71.7 (CH), 68.3 (Cq), 67.4 (CH.sub.2), 58.4 (CH), 52.4 (CH.sub.2), 29.1 (CH.sub.2), 22.0 (CH.sub.3) ppm; HRMS (ESI.sup.+) Calc. C.sub.9H.sub.18NO.sub.4 [M+H].sup.+: m/z=204.12303; Found m/z=204.12232.
Compound (IIz4)
[0347] The compound of above formula (IIz4) ((3S,4R,5R,6R,8S)-8-cyclopentyl-6-(hydroxymethyl)-1-azabicyclo[4.2.0]octane-3,4,5-triol: 14.6 mg; 62% for 2 steps) was prepared in accordance with the general procedures C and B, from β-lactam C-5 (70.0 mg; 0.124 mmol) and 2M cyclopentylmagnesium chloride in ether (R.sup.14=cPent; 36.4 μL; 0.248 mmol).
[0348] A yellowish solid is obtained, with the following characteristics:
[0349] .sup.1H NMR (500 MHz, CD.sub.3OD) δ 3.98 (t, J=7.0 Hz, 1H), 3.76-3.63 (m, 2H), 3.58-3.36 (m, 3H), 3.10 (dd, J=14.5, 5.0 Hz, 1H), 2.76 (dd, J=14.5, 4.0 Hz, 1H), 2.50 (t, J=11.8 Hz, 1H), 2.06-1.95 (m, 1H), 1.89-1.76 (m, 1H), 1.74-1.47 (m, 6H), 1.36-1.22 (m, 1H), 1.15-1.00 (m, 1H) ppm;
[0350] .sup.13C NMR (125 MHz, CD.sub.3OD) δ 77.7 (CH), 75.0 (CH), 71.9 (CH), 67.5 (CH.sub.2), 67.2 (CH), 66.3 (Cq), 54.2 (CH.sub.2), 48.9 (CH), 31.0 (CH.sub.2), 29.1 (CH.sub.2), 27.0 (CH.sub.2), 26.4 (CH.sub.2), 25.9 (CH.sub.2) ppm;
[0351] HRMS (ESI.sup.+) Calc. C.sub.13H.sub.24NO.sub.4 [M+H].sup.+: m/z=258.16998; Found m/z=258.16987.
Compound (IIz5)
[0352] The compound of above formula (IIz5) ((3S,4R,5R,6R,8R)-6-(hydroxymethyl)-8-pentyl-1-azabicyclo[4.2.0]octane-3,4,5-triol: 19.1 mg; 45% for 2 steps) was prepared in accordance with the general procedures C and B, from β-lactam C-5 (70.0 mg; 0.124 mmol) and 2M pentylmagnesium chloride in THF (R.sup.14=n-Pent; 33.6 μL; 0.248 mmol).
[0353] A yellow solid is obtained, with the following characteristics:
[0354] [α].sup.20.sub.D−31.1 (c 0.46, MeOH);
[0355] 1H NMR (500 MHz, CD.sub.3OD) δ 3.99 (t, J=7.0 Hz, 1H), 3.74-3.64 (m, 2H), 3.64-3.55 (m, 1H), 3.51 (d, J=11.1 Hz, 1H), 3.46 (d, J=11.1 Hz, 1H), 3.09 (dd, J=13.7, 5.4 Hz, 1H), 2.70 (dd, J=13.7, 4.0 Hz, 1H), 2.52 (dd, J=11.4, 8.9 Hz, 1H), 1.68-1.53 (m, 2H), 1.51-1.39 (m, 1H), 1.39-1.21 (m, 6H), 0.90 (t, J=6.7 Hz, 3H) ppm;
[0356] .sup.13C NMR (100 MHz, CD.sub.3OD) δ 77.5 (CH), 75.2 (CH), 71.9 (CH), 67.8 (CH.sub.2), 66.4 (Cq), 62.4 (CH), 53.8 (CH.sub.2), 38.8 (CH.sub.2), 33.0 (CH.sub.2), 27.9 (CH.sub.2), 26.5 (CH.sub.2), 23.7 (CH.sub.2), 14.3 (CH.sub.3) ppm;
[0357] HRMS (ESI.sup.+) Calc. C.sub.13H.sub.26NO.sub.4 [M+H].sup.+: m/z=260.18563; Found m/z=260.18561.
A.6/ Synthesis Method 7
[0358] The synthesis method used for obtaining the compounds complying with the general formula (I′c) below falls under the general synthesis scheme 7 shown on
Compound (IIk)
[0359] The compound of formula (IIk) as above ((3S,4R,5R,6R,7S)-6,7-bis(hydroxymethyl)-1-azabicyclo[4.1.0]heptane-3,4,5-triol) is obtained in the following manner.
[0360] A mixture of nitrone (III) (P=Bn; 193 mg, 0.360 mmol) and of alkyne Vc (R.sup.8=NBnTs; 308 mg, 1.079 mmol) in solution in CH.sub.2Cl.sub.2 (0.4 mL) was stirred at ambient temperature for 3 days, and then the reaction mixture was concentrated under reduced pressure. The residue (raw cycloadduct B-7c) was dissolved in ethanol (0.1M), transferred into a sealed tube, and heated at 110° C. (IR probe) under microwave irradiation for 15 minutes. After concentration under reduced pressure, the residue was purified by chromatography to give acylaziridine H-7c (P=Bn, R.sup.8=NBnTs; 238 mg, 80% on 2 steps). A fraction of this compound (58 mg, 0.070 mmol) was dissolved in THF (1 mL), and then LiAlH4 (6 mg, 0.157 mmol) was added at 0° C. The mixture was stirred for 1.5 hours at ambient temperature. After adding water (0.1 mL) and a 10% aqueous solution of NaOH (0.1 mL), the reaction mixture was stirred at ambient temperature for 2 hours and then filtered over Celite®. The filtrate, concentrated under reduced pressure, was purified by chromatography to isolate the aziridine-alcohol J-7c (P=Bn, R.sup.16=H; 34.2 mg, 86%). A solution of this aziridine-alcohol J-7c (28 mg, 0.067 mmol) in THF (2 mL) was added to a solution of lithium (54 mg, 7.78 mmol) in liquid ammonia (10 mL) at −78° C. The mixture was stirred for 45 minutes at -78° C., and then Milli-Q water (0.5 mL) and MeOH (2 mL) were added. After returning the solution to ambient temperature, the solvents were evaporated under reduced pressure. The residue was dissolved in Milli-Q water and neutralized with Amberlite® IR-120 (22.8 g) previously treated with 1M HCl (15 mL). The resin was introduced into a column, washed with water, and then the compound (IIk) was eluted with a 1M solution of aqueous NH.sub.4OH. The solution obtained after evaporation of the solvents under reduced pressure was purified by chromatography to give the pure iminosugar (IIk) (R.sup.16=H; 7 mg, 60%).
[0361] A white solid is obtained, with the following characteristics:
[0362] [α].sup.20.sub.D=+2.90 (c 0.34, CH.sub.3OH);
[0363] IR v 3409, 3010, 2926, 1084, 1032 cm.sup.−1;
[0364] .sup.1H NMR (500 MHz, CD.sub.3OD) δ 4.16 (d, J=7.9 Hz, 1H), 3.79 (d, J=11.6 Hz, 1H), 3.60-3.50 (m, 3H), 3.47-3.40 (m, 1H), 3.33-3.25 (m, 2H), 2.55 (dd, J=10.6, 12.4 Hz, 1H), 2.19 (dd, J=6.8, 5.3 Hz, 1H) ppm;
[0365] .sup.13C NMR (125 MHz, CD.sub.3OD) 576.0 (CH), 73.2 (CH), 70.6 (CH), 63.2 (CH.sub.2), 62.0 (CH.sub.2), 57.0 (CH.sub.2), 51.9 (C.sub.q), 51.6 (CH) ppm;
[0366] HRMS (ESI.sup.+) Calc. C.sub.8H.sub.16NO.sub.5 [M+H].sup.+ m/z=206.1028; Found m/z=206.1028.
Compound (IIj)
[0367] The compound of above formula (IIj) ((3S,4R,5R,6R,7S)-7-((S)-1,2-dihydroxyethyl)-6-(hydroxymethyl)-1-azabicyclo[4.1.0]heptane-3,4,5-triol) is obtained as follows.
[0368] Acylaziridine H-7d (P=Bn, R.sup.8=CH.sub.2OAc; 33 mg, 0.052 mmol), obtained as described in the publication by Tangara et al., 2017, Org. Lett. 19: 4842-4845, was dissolved in THF (1 mL), and then LiAlH4 (38 mg, 0.165 mmol) was added at 0° C. and the mixture was stirred for 1 hour at ambient temperature. After adding water (0.1 mL) and a 10% aqueous solution of NaOH (0.1 mL), the reaction mixture was stirred at ambient temperature for 2 hours and then filtered over Celite®. The filtrate, concentrated under reduced pressure, was purified by chromatography to isolate a translucent oil J-7d (R.sup.16=CH.sub.2OH; 26 mg, 83%). A solution of this aziridine-alcohol J-7d (40 mg, 0.067 mmol) in THF (2 mL) was added to a solution of lithium (25 mg, 3.571 mmol) in liquid ammonia (10 mL) at −78° C. The mixture was stirred for 45 minutes at −78° C. and then Milli-Q water (0.5 mL) and MeOH (2 mL) were added. After returning the solution to ambient temperature, these solvents were evaporated under reduced pressure. The residue was dissolved in Milli-Q water and neutralized with Amberlite® IR-120 (8.5 g) previously treated with 1M HCl (5 mL). The resin was introduced into a column, washed with water, and then the compound (IIj) was eluted with a 1M solution of aqueous NH.sub.4OH. The solid obtained after evaporation of the solvents under reduced pressure was purified by chromatography to give the pure iminosugar (IIj) (R.sup.16=CH.sub.2OH; 10.1 mg, 64%).
[0369] A white solid is obtained, with the following characteristics:
[0370] [α].sup.20.sub.D=+5.90 (c 0.34, CH.sub.3OH);
[0371] IR: v 3249, 2939, 2885, 1748, 1656, 1404, 1031, 995 cm.sup.−1;
[0372] .sup.1H NMR (500 MHz, CD.sub.3OD) δ 4.16 (d, J=8.0 Hz, 1H), 3.91 (d, J=11.6 Hz, 1H), 3.71 (dd, J=3.5, 11.4 Hz, 1H), 3.61 (dd, J=5.8, 11.4 Hz, 1H), 3.54-3.48 (m, 1H), 3.48-3.41 (m, 1H), 3.39 (d, J=11.6 Hz, 1H), 3.30-3.25 (m, 2H), 2.52 (dd, J=10.2, 12.2 Hz, 1H), 2.10 (d, J=8.7 Hz, 1H) ppm;
[0373] .sup.13C NMR (125 MHz, CD.sub.3OD) δ 75.9 (CH), 73.1 (CH), 72.4 (CH), 70.7 (CH), 66.1 (CH.sub.2), 63.8 (CH.sub.2), 56.5 (CH.sub.2), 52.0 (C.sub.q), 51.2 (CH) ppm;
[0374] HRMS (ESI.sup.+) Calc. C.sub.9H.sub.18NO.sub.6 [M+H].sup.+m/z=236.1129; Found m/z=236.1124.
Compound (IIm)
[0375] The compound of above formula (IIm) (3S,4R,5R,6R,7S)-7-((S)-hydroxy(phenyl)methyl)-6-(hydroxymethyl)-1-azabicyclo[4.1.0]heptane-3,4,5-triol is obtained as follows.
[0376] The cycloadducts B-7e (P=Ac, R.sup.8=Ph; 15.2 mg, 0.030 mmol, 4:1 mixture of diastereoisomers), obtained as described in the publication by Tangara et al., 2018, New J. Chem. 42: 16735-16743, were dissolved in dichloroethane (0.1M solution) in a sealed tube. The solution was heated at 110° C. (IR probe) under irradiation by microwave for 45 minutes (30+15). After evaporation of the solvent under reduced pressure, purification of the residue by chromatography made it possible to isolate the acylaziridines H-7e (P=Ac, R.sup.8=Ph; 12 mg, 79%) in the form of a 9:1 mixture of two diastereoisomers. These acylaziridines (30 mg, 0.067 mmol) were dissolved in THF (1.2 mL), the solution was cooled to 0° C. and then LiAlH4 (5.6 mg, 0.147 mmol) was added. The reaction mixture was stirred at ambient temperature for 1.5 hours, and then water (0.1 mL) and a 10% aqueous solution of NaOH (0.1 mL) were added thereto. After stirring at ambient temperature for 1.5 hours, this mixture was filtered over Celite®, the filtrate being concentrated under reduced pressure, and the residue was purified by chromatography to provide the aziridinyl iminosugar (IIm) (R.sup.16=Ph; 15 mg, 82%) in the form of a single diastereoisomer.
[0377] A colorless oil is obtained, with the following characteristics:
[0378] [α].sup.20.sub.D−4.1 (c 1.58, CH.sub.3OH);
[0379] IR v 3296, 2931, 1557, 1409, 1053, 697 cm.sup.−1;
[0380] .sup.1H NMR (500 MHz, CD.sub.3OD) δ 7.49-7.44 (m, 2H), 7.40-7.35 (m, 2H), 7.31-7.25 (m, 1H), 4.29 (d, J=8.7 Hz, 1H), 4.18 (d, J=7.9 Hz, 1H), 4.01 (d, J=11.6 Hz, 1H), 3.56 (d, J=11.6 Hz, 1H), 3.48-3.37 (m, 2H), 3.26 (dd, J=9.1, 8.0 Hz, 1H), 2.26-2.18 (m, 2H) ppm;
[0381] .sup.13C NMR (125 MHz, CD.sub.3OD) δ 144.6 (.sup.ArC.sub.q), 129.5 (.sup.ArCH), 128.6 (.sup.ArCH), 127.1 (.sup.ArCH), 75.8 (CH), 73.8 (CH), 73.1 (CH), 70.7 (CH), 63.6 (CH.sub.2), 56.4 (CH.sub.2), 56.0 (CH), 52.8 (C.sub.q)
[0382] ppm;
[0383] HRMS (ESI.sup.+) Calc. C.sub.14H.sub.20NO.sub.5 [M+H].sup.+: m/z=282.1341; Found m/z=282.1346.
Compound (IIn)
[0384] The compound of above formula (IIn) (3S,4R,5R,6R,7S)-7-((R)-cyclohexyl(hydroxy)methyl)-6-(hydroxymethyl)-1-azabicyclo[4.1.0]heptane-3,4,5-triol is obtained as follows.
[0385] Acylaziridine H-7f (P=Bn, R.sup.8=c-Hex; 77 mg, 0.119 mmol), obtained as described in the publication by Tangara et al., 2017, Org. Lett. 19: 4842-4845, was dissolved in ethanol (1.4 mL), cooled to 0° C., and then NaBH.sub.4 (13 mg, 0.342 mmol) was added. The mixture was stirred at the same temperature for 3.3 hours, and then water, a 10% aqueous solution of NaOH and CH.sub.2Cl.sub.2 were added. The aqueous phase was extracted (3 times) by CH.sub.2Cl.sub.2, the organic phases were collected, dried on MgSO.sub.4 and concentrated under reduced pressure. The residue thus obtained was filtered on silica to isolate the aziridine-alcohol J-7f (R.sup.16=c-Hex; 72 mg, 94%) in the form of an oil. A solution of this aziridine-alcohol J-7f (42 mg, 0.064 mmol) in THF (4 mL) was added to a solution of lithium (12 mg, 2.0 mmol) in liquid ammonia (10 mL) at −78° C. The reaction mixture was stirred for 45 minutes at −78° C., and then Milli-Q water (0.5 mL) and MeOH (2 mL) were added. After returning the solution to ambient temperature, the solvents were evaporated under reduced pressure. The residue was dissolved in Milli-Q water and neutralized with Amberlite® IR-120 (3.5 g) previously treated with 1M HCl (2 mL). The resin was introduced into a column, washed with water, and then the compound (IIn) was eluted with a 1M solution of aqueous NH.sub.4OH. The solid obtained after evaporation of the solvents under reduced pressure was purified by chromatography to give the pure iminosugar (IIn) (R.sup.16=c-Hex; 13 mg, 72%).
[0386] A white solid is obtained, with the following characteristics:
[0387] [α].sup.20.sub.D+2.8 (c 0.80, CH.sub.3OH);
[0388] IR v 3335, 2916, 2846, 1451, 1087, 1054, 1008, 660 cm.sup.−1;
[0389] .sup.1H NMR (500 MHz, CD.sub.3OD) δ 4.17 (d, J=7.8 Hz, 1H), 3.88 (d, J=11.7 Hz, 1H), 3.50-3.44 (m, 2H), 3.42 (d, J=11.7 Hz, 1H), 3.33-3.29 (m, 1H), 2.95 (dd, J=8.6, 6.6 Hz, 1H), 2.51 (dd, J=11.8, 9.0 Hz, 1H), 2.08 (d, J=8.7 Hz, 1H), 1.99-1.66 (m, 5H), 1.53-1.45 (m, 1H), 1.36-1.03 (m, 5H) ppm;
[0390] .sup.13C NMR(125 MHz, CD.sub.3OD) δ 76.3 (CH), 76.0 (CH), 73.3 (CH), 70.8 (CH), 64.1 (CH.sub.2), 56.0 (CH.sub.2), 52.9 (CH), 51.1 (C.sub.q), 45.3 (CH), 30.1 (CH.sub.2), 29.7 (CH.sub.2), 27.7 (CH.sub.2), 27.4 (CH.sub.2), 27.2 (CH.sub.2) ppm;
[0391] HRMS (ESI.sup.+) Calc. C.sub.14H.sub.26NO.sub.5 [M+H].sup.+: m/z=288.1811; Found m/z=288.1810.
A.7/ Synthesis Method 8
[0392] A synthesis method used for obtaining a compound complying with the general formula (I′e) below falls under the general synthesis scheme 8 shown on
Compound (IIw1)
[0393] The compound of above formula (IIw1) ((2R,3R,4R,5S)-2-(hydroxymethyl)-2-(1H-1,2,3-triazol-4yl)piperidine-3,4,5-triol is prepared in the following manner. A solution of hydroxylamine of formula A-1a (240.0 mg; 0.377 mmol) and of zinc powder (247.0 mg; 3.77 mmol) in a 4:1 mixture EtOH/AcOH (5 mL) was stirred at 65° C. under ultrasound until the reaction was complete. The reaction mixture is filtered over Celite and then evaporated under reduced pressure. The raw product is redissolved in CH.sub.2Cl.sub.2 and then treated with aqueous 1M NaOH. The aqueous phase was extracted (three times) with CH.sub.2Cl.sub.2. The organic phases were washed with brine, dried over MgSO.sub.4 and then evaporated under reduced pressure. Purification of the residue obtained by chromatography provided the corresponding piperidine D-8 (178.0 mg; 76%). A mixture of this piperidine D-8 (176.0 mg; 0.284 mmol), of benzyl chloroformiate (121 μL; 0.852 mmol) and of K.sub.2CO.sub.3 (157.0 mg; 1.14 mmol) in anhydrous THF (1.7 mL) was stirred at ambient temperature until the reaction was complete. Methanol (2 mL) was added and then the reaction mixture was stirred at ambient temperature until the reaction was complete. Purification of the residue obtained by chromatography provided the compound of formula K-8 (164.0 mg; 78% over 2 steps). Copper iodide (3.5 mg; 0.018 mmol) and DIPEA (108 μL; 0.618 mmol) were added to a solution of this same compound K-8 (46.8 mg; 0.062 mmol) and of benzyl azide (247 μL; 0.124 mmol) in DMF (1.5 mL) under inert atmosphere. The reaction mixture was stirred at ambient temperature until the reaction was complete. The raw product was diluted in ethyl acetate and washed several times with brine. The organic phase was dried over MgSO.sub.4 and evaporated at reduced pressure. Purification of the residue obtained by chromatography provided the compound of formula L-8 (38.2 mg; 76%). This compound (32.1 mg; 0.039 mmol) was debenzylated in accordance with the general procedure B to give piperidine (IIw1) (11.0 mg; 100%).
[0394] A colorless lacquer is obtained, with the following characteristics:
[0395] [α].sup.20.sub.D−17.2 (c 1.09, CH.sub.3OH);
[0396] 1H NMR (500 MHz, CD.sub.3OD) δ 7.90 (s, 1H), 3.80 (d, J=10.9 Hz, 1H), 3.69 (d, J=9.7 Hz, 1H), 3.59 (d, J=11.1 Hz, 1H), 3.52-3.45 (m, 1H), 3.27 (t, J=9.3 Hz, 1H), 2.99 (dd, J=12.6, 5.5 Hz, 1H), 2.54 (dd, J=12.3, 11.2 Hz, 1H) ppm;
[0397] .sup.13C NMR (125 MHz, CD.sub.3OD) δ 144.5 (Cq), 131.0 (CH), 76.9 (CH), 74.5 (CH), 73.0 (CH), 68.1 (CH.sub.2), 62.5 (Cq), 47.2 (CH.sub.2) ppm;
[0398] HRMS (ESI.sup.+) Calc. C.sub.8H.sub.15N.sub.4O.sub.4 [M+H].sup.+: m/z=231.10878; Found m/z=231.10861.
B/ Biological Evaluations
[0399] The following various biological activity tests are implemented on the compounds (IIa) to (IIn) described above.
B.1/ Inhibition of rhGAA Determined by Fluopol-ABPP
[0400] The inhibiting activity on recombinant human acid α-glucosidase (rhGAA) of the compounds is implemented using the Fluopol-ABPP method (Fluorescence Polarization Activity Based Protein Profiling) described in the publication of Lahav et al., 2017, J. Am. Chem. Soc. 139: 14192-14197. This technique, based on the competition between an inhibitor and a fluorescent probe capable of binding covalently to the active site of an enzyme, makes it possible to measure the affinity of this inhibitor for the active site of the rhGAA enzyme used for these experiments by the laboratory of Professors Herman S. Overkleeft and Johanes M. F. G. Aerts, Leiden Institute of Chemistry, Leiden University (NL), is the enzyme marketed under the name Myozyme®. The median inhibiting concentrations (IC.sub.50) are determined in the Mcllvaine buffer (citrate-phosphate) 150 mM at pH 5.0, in the presence of 0.1% bovine gamma-globulin (p/v) and 0.5 mg/mL of Chaps detergent (Sigma) in 96-well plates (Griener). The rhGAA enzyme (10 μg/mL) is pre-incubated with solutions of inhibitor (containing 2.5% DMSO that was used to prepare the mother solutions of the compounds) at various concentrations [I] in the buffer, for 45 minutes at 37° C. The tetraaminomethylrhodamine (TAMRA) fluorescent probe in solution (25 nM) in the buffer is next added to the mixture. After 4 hours, the samples are irradiated with a polarized light (λ=530 nm) and the fluorescence emitted (λ=580 nm) is measured by means of an Infinite® M1000Pro (Tecan) spectrofluorimeter. For each concentration of inhibitor, the percentage of inhibition of the enzyme is determined by the formula:
%Inhibition=[F.sub.measured−F.sub.control1)/F.sub.contro2)×100
where F.sub.measured corresponds to the fluorescence measured in the presence of the iminosugars; F.sub.control1 corresponds to the fluorescence measured in the presence of a powerful inhibitor of human acid α-glucosidase acid serving as a positive control (100% inhibition), CF 022 ((1S,2R,3S,4R,5R,6R)-2,3,4-trihydroxy-5-(hydroxymethyl)-7-(8-azidooctyl)-7-aza-bicyclo[4.1.0]heptane), and F.sub.control2 represents the fluorescence of the probe measured in the absence of the inhibitor (0% inhibition).
[0401] The IC.sub.50 values are calculated by a non-linear regression of the % inhibition as a function of the concentration [I] by means of the GraphPad Prism 6.0 software. The results are a mean of three identical experiments (triplicates).
B.2/ Inhibition of rhGAA Determined by Measuring the Residual Activity in the Presence of an Inhibitor in Vitro
[0402] In a complementary way, the activity of inhibition of recombinant human acid α-glucosidase (rhGAA) of the compounds was evaluated by the team of Professor Marco Moracci, Department of Biology, University of Naples Federico II (IT), using the method described in the publication of Porto et al., 2012, Molecular Therapy 20: 2201-2211. The rhGAA enzyme sold under the name Myozyme® used comes from residues of perfusions of the recombinant enzyme used for treating, by enzyme therapy, the patients with Pompe disease in the Department of Translational Medical Sciences, University of Naples Federico II (IT). The compounds are solubilized at various concentrations in a 100 mM sodium acetate buffer, pH 4.0, as well as the 4-nitrophenyl-α-
B.3/ Determination of the Thermal Stabilization of rhGAA
[0403] The stabilization of rhGAA in the presence of the compounds is determined in accordance with the method described in the publication of Niesen et al., 2007, Nat. Protoc. 2: 2212-2221. The rhGAA enzyme (2.5 μg, 0.1 mg/mL) is incubated in the absence and in the presence of each compound (at a concentration [I]=100 μM), of SYPRO® orange dye (Life Technologies), and of sodium phosphate buffer (25 mM) and of NaCI (150 mM), pH 7.4 or of sodium acetate buffer (25 mmol) and of NaCl (150 mM), pH 4.0. The thermal stability of the enzyme in these various conditions is evaluated by DSF (differential scanning fluorimetry) by varying the temperature by 1° C./min over an interval of 25 to 95° C., and measuring the fluorescence of the SYPRO® orange dye every minute by means of a Real-Time Cycler spectrofluorimeter (Biorad). The relative fluorescence is determined by comparing each fluorescence value measured with that of the maximum fluorescence value of the SYPRO® orange dye for each scan. The results are a mean of three identical experiments (triplicates).
B.4/ Evaluation of the Inhibition of Other Enzymes of Human Origin
[0404] The selectivity of the compounds was evaluated in the laboratory of Professors Herman S. Overkleeft and Johanes M. F. G. Aerts, Leiden Institute of Chemistry, Leiden University (NL). The enzymes used for determining the selectivity of inhibition of the compounds with respect to various human enzymes are the α-glucosidase II of the endoplasmic reticulum (GANAB), recombinant human lysosomal β-glucocerebrosidase (GBA1), human non-lysosomal β-glucosylceramidase (GBA2) and β-glucosylceramide synthase (GCS). The inhibiting activity of the compounds on these various enzymes is determined as described in the publication by Artola et al., 2017, ACS Cent. Sci. 3: 784-793. The enzyme GBA1 sold under the name Cerezyme® and the enzyme rhGAA sold under the name Myozyme® are used. The human GANAB enzyme used for this study is that of fibroblasts of patients suffering from Pompe disease diagnosed as lacking active GAA and voluntary donors. These fibroblasts were cultivated on an HAMF12-DMEM medium (Sigma) supplemented by 10% (v/v) FCS (fetal calf serum). The GBA2 enzyme was over-expressed in HEK298T cells cultivated on a DMEM medium enriched with glucose (Gibco) supplemented by 10% NBS (newborn bovine serum) and 100 units/mL of penicillin/streptomycin (Gibco) and 5% CO.sub.2 at 37° C. The activity of the compounds on human β-glucosylceramide synthase (GCS) is evaluated in situ on cells of the RAW 264.7 type cultivated on an RPMI medium (Gibco) supplemented by 10% FCS, 1 mM of glutamax, 100 units/mL of penicillin/streptomycin (Gibco) and 5% CO.sub.2 at 37° C.
[0405] The IC.sub.50 values for the GANAB, GBA2 and GCS enzymes are determined from cell lysates prepared in a buffer (20 mM hepes, 2 mM DTT, 0.25 M sucrose, 1 mM MgCl.sub.2, 2.5 U/mL benzonase) at pH 7.0, and placed on ice for 30 minutes. These cell lysates are homogenized using a SilentCrusher grinder (Heidolph®), and then subjected to ultracentrifugation at 32,000 rpm for 30 minutes at 4° C. The total protein concentration is determined in accordance with the Bradford method (Bradford, 1976, Anal. Biochem. 72: 248-254), using a Bradford BioRad Quick Start® kit (Pierce) and BSA (Sigma). The lysates are next aliquoted and stored at −80° C. before use.
[0406] The IC.sub.50 values for the GANAB enzyme are determined on cell lysates of fibroblasts of patients suffering from Pompe disease, using as a buffer the Mcllvaine 150 mM, pH 7.0, 0.1% bovine serum albumin (BSA) (p/v), a substrate (4-methyl-umbelliferone-α-
[0407] The IC.sub.50 values for the GBA1 enzyme are determined using as buffer the Mcllvaine 150 mM, pH 5.2, 0.2% taurocholate (p/v), 0.1% Triton X-100 (v/v), 0.1% bovine serum albumin (BSA) (p/v), an enzyme concentration of 0.7 nM, a substrate (4-methyl-umbelliferone-β-
[0408] The residual activity of GBA2 in the presence of the compounds is determined after pre-incubation for 30 minutes of the homogenates of HEK298T cells over-expressing GBA2 with an inhibitor of GBA1, conduritol β-epoxide (Sigma), at a concentration of 1 mM. The IC.sub.50 values are determined on cell lysates, using as a buffer the Mcllvaine 150 mM, pH 5.8, 0.1% bovine serum albumin (BSA) (p/v), a substrate (4-methyl-umbelliferone-β-
[0409] The residual activity of GCS in the presence of the compounds is determined after pre-incubation for 1 hour of the homogenates of RAW 264.7 cells with an inhibitor of GBA1, conduritol β-epoxide (Sigma), at a concentration of 300 μM.
[0410] The I.sub.50 values for the GCS enzyme are determined in situ in the cell culture medium at pH 7.0 using 1 μM of NBD-ceramide (N-[12-[(7-nitro-2-1,3-benzoxadiazol-4-yl)amino]dodecanoyl]-D-erythro-sphingosine) as substrate, as described in the aforementioned publication of Lahav et al.
[0411] The results obtained for all these tests are set out in tables 1 and 2 below. In these tables, by way of comparison, the values obtained for the compounds of the prior art DNJ and NB-DNJ, as described in the literature, are also indicated. In these tables: .sup.a indicates Porto et al., 2012, Mol. Ther. 20: 2201-2211; .sup.b indicates Bruckmann et al., 2012, ChemMedChem 7: 1943-1953; .sup.c indicates Flanagan et al., 2009, Human Mut. 30: 1683-1692; .sup.d indicates D'Alonzo et al., 2017, J. Med. Chem. 60: 9462-9469; * indicates ΔTm measured at [compound]=10 times the value of K.sub.i; .sup.e indicates Asano et al., 1995, J. Med. Chem. 38: 2349-56; .sup.f indicates Wennekes et al., 2010, J. Med. Chem. 53: 689-698.
TABLE-US-00001 TABLE 1 rhGAA rhGAA rhGAA rhGAA (Fluopol- (residual ΔTm ΔTm Com- ABPP) activity) at pH at pH pound IC.sub.50 (μM) IC.sub.50 (μM) 4.0 (° C.) 7.4 (° C.) (IIa) 0.42 341 8.1 12.7 (IIb) 0.39 104 7.8 11.5 (Ki = 13 μM) (IIc) 0.22 37.2 11.8 13.0 (K.sub.i = 7.6 μM) (IId) 0.15 182 9.4 10.7 (K.sub.i = 18.9 μM) (IIf) 0.87 297 6.4 11.8 (IIg) 0.46 387 6.7 14.4 (IIh) 9.7 — — — (IIj) 15.2 — — — (IIk) 11.6 — — — (IIm) 45.0 — — — (IIn) 24.6 — — — (IIx1) — 40.3 — 16.5 (IIb7) — 31.6 — 16.8 (IIb3) — 29.6 — 16.9 (K.sub.i = 10.8 μM) (IIc1) — 32.9 — 15.0 (K.sub.i = 13.0 μM) (IIr) — 29.5 — 14.5 (K.sub.i = 20.0 μM) (IIb1) — 49.1 — 6.2 (K.sub.i = 6.4 μM) DNJ 0.41 K.sub.i = 3.4 μM.sup.a 2.7 (pH 4.3)*.sup.b 15.sup.c.sup. NB-DNJ — K.sub.i = 3.1 μM.sup.b 6.2 (pH 4.3)*.sup.b 12.sup.d.sup.
TABLE-US-00002 TABLE 2 GANAB GBA1 GBA2 GCS Compound IC.sub.50 (μM) IC.sub.50 (μM) IC.sub.50 (μM) IC.sub.50 (μM) (IIa) >100 >1000 >1000 >50 (IIb) >100 >1000 >1000 >50 (IIc) >100 >1000 >1000 >50 (IId) >100 >1000 >1000 >50 (IIf) >100 >1000 >1000 >50 (IIg) >100 >1000 >1000 >50 DNJ .sup. 4.6.sup.e 250.sup.f 21 >100.sup.f NB-DNJ .sup. 15.sup.e 400.sup.f 0.23.sup.f 50.sup.f
[0412] All these results demonstrate a selectivity of the compounds used in accordance with the invention for human acid α-glucosidase, compared with the other human enzymes tested. This selectivity is much greater than that of the molecules proposed by the prior art DNJ and NB-DNJ. The compounds used in accordance with the invention furthermore have an important effect of stabilization of rhGAA, comparable to that of DNJ and NB-DNJ.
[0413] Furthermore, the compounds (IIb7), (IIb3), (IIc1), (IIr) and (Ic), complying with the general formula (I″a), as well as the compound (IIx1), have performances superior to those of the other compounds according to the invention.