Patent Application
20120282180
This application claims priority to European patent application number EP 11 165 157.6, filed on May 6, 2011; the entire contents of which is incorporated by reference.
The present invention relates to the field of therapeutic and diagnostic agents and more specifically to compounds that are inhibitors of matrix-metalloproteinases (MMPs) and are useful in the treatment of diseases related thereto such as cardiovascular diseases, inflammatory diseases and malignant diseases. The compounds are also useful in the field of in vivo diagnostic imaging and in particular in (Positron Emission Tomography) PET imaging.
The matrix-metalloproteinases (MMPs) are a family of at least 20 zinc-dependent endo-peptidases which mediate degradation, or remodeling of the extracellular matrix (ECM) [Massova et al., FASEB J. 1998, 12, 1075-1095]. Together, the members of the MMP family can degrade e.g. components of the blood vessel wall and play a major role in both physiological and pathological events that involve the degradation of components of the ECM. Since the MMPs can interfere with the cell-matrix interactions that control cell behavior, their activity affects processes as diverse as cellular differentiation, migration, proliferation and apoptosis [Nagase and Woessner, J. Biol. Chem. 1999, 274, 21491-21494]. The negative regulatory controls that finely regulate MMP activity in physiological situations do not always function as they should. Inappropriate expression of MMP activity is thought to constitute part of the pathological mechanism in several disease states. MMPs are therefore targets for therapeutic inhibitors in many inflammatory, malignant and degenerative diseases [Whittaker et al., Chem. Rev. 1999, 99, 2735-2776].
Consequently, it is believed that synthetic inhibitors of MMPs may be useful in the treatment of many inflammatory, malignant and degenerative diseases. Furthermore, it has been suggested that inhibitors of MMPs may be useful in the diagnosis of these diseases. WO 01/60416 discloses compounds, which are proposed to be useful in the diagnosis of cardiovascular pathologies associated with extracellular matrix degradation such as atherosclerosis, heart failure and restenosis. The compounds disclosed therein comprise MMP inhibitors linked, via an optional linker, to a chelator capable of conjugating to a diagnostic metal. Preferred MMP inhibitors, chelators and linkers are described therein. A report by Zheng et al. [Nuc. Med. Biol. 2002, 29, 761-770] documented the synthesis of MMP inhibitors labeled with the positron emission tomography (PET) tracers 11C and 18F. These compounds described therein are useful in the non-invasive imaging of breast cancer.
Novel compounds having MMP inhibitory activity are disclosed, which have been found to be particularly useful in the prevention, treatment and diagnostic imaging of diseases associated with an unpaired activity of MMP, amongst others MMP-2, MMP-8, MMP-9 and/or MMP-13.
Another aspect of the present invention relates to pharmaceutical compositions useful in prevention, treatment and diagnostic imaging of diseases associated with an unpaired activity of MMP.
The compounds of the present invention are useful for the prevention, the treatment and the in vivo diagnostic imaging of a range of disease states (inflammatory, malignant and degenerative diseases) where specific matrix metalloproteinases are known to be involved. These include:
(a) atherosclerosis, where various MMPs are overexpressed. Elevated levels of MMP-1, 3, 7, 9, 11, 12, 13 and MT1-MMP have been detected in human atherosclerotic plaques [George, Exp. Opin. Invest. Drugs 2000, 9, 993-1007 and references therein]. Expression of MMP-2 [Li et al. Am. J. Pathol. 1996, 148, 121-128] and MMP-8 [Herman et al., Circulation 2001, 104, 1899-1904] in human atheroma has also been reported;
(b) CHF [Peterson et al., Matrix metalloproteinase inhibitor development for the treatment of heart failure, Drug Dev. Res. 2002, 55, 29-44] report that MMP-1, MMP-2, MMP-3, MMP-8, MMP-9, MMP-13 and MMP-14 are upregulated in heart failure;
(c) cancer [Vihinen et al., Int. J. Cancer 2002, 99, 157-186] reviews MMP involvement in cancers, and particularly highlights MMP-2, MMP-3, MMP-7, and MMP-9];
(d) arthritis [Meson et al., Inflamm. Res. 2001, 50, 183-186 “Selective matrix metalloproteinase inhibition in rheumatoid arthritis-targeting gelatinase A activation”], MMP-2 is particularly discussed;
(e) amyotrophic lateral sclerosis [Lim et al., J. Neurochem. 1996, 67, 251-259] where MMP-2 and MMP-9 are involved;
(f) brain metastases, where MMP-2, MMP-9 and MMP-13 have been reported to be implicated [Spinale, Circul. Res. 2002, 90, 520-530];
(g) cerebrovascular diseases, where MMP-2 and MMP-9 have been reported to be involved [Lukes et al., Mol. Neurobiol. 1999, 19, 267-284];
(h) Alzheimer's disease, where MMP-2 and MMP-9 have been identified in diseased tissue [Backstrom et al., J. Neurochem. 1992, 58, 983-992];
(i) neuroinflammatory diseases, where MMP-2, MMP-3 and MMP-9 are involved [Mun-Bryce et al., Brain. Res. 2002, 933, 42-49];
(j) COPD (i.e. chronic obstructive pulmonary disease) where MMP-1, MMP-2, MMP-8 and MMP-9 have been reported to be upregulated [Segura-Valdez et al., Chest. 2000, 117, 684-694];
(k) eye pathology [Kurpakus-Wheater et al., Prog. Histo. Cytochem. 2001, 36, 179-259];
(l) skin diseases [Herouy, Int. J. Mol. Med. 2001, 7, 3-12].
It has been surprisingly found that compounds of below general formula (I) wherein the group R3 is an alkenyl optionally substituted with a fluorine or an alkyl substituted with a fluorine act as inhibitors of MMP and in particular of MMP-2 and MMP-9. It has been surprisingly found that the presence of the group R3 as defined below in formula (I) provide these compounds with a very good inhibitory activity. Additionally, it has been surprisingly found that the inhibition is remarkable also in the (S)-enantiomer of the compounds of formula (I). This is particularly surprising because the (R)-enantiomer of the prior art analog compounds (e.g. hydroxamate derivatives such as CGS 27023A) are far more active than the corresponding (S)-enantiomers. For example, it is known that the (S)-enantiomer of compound CGS 27023A is far less potent then the corresponding (R)-enantiomer in the inhibition of MMP-2 and MMP-9 (see table 1). As a consequence, the use of the (S)-enantiomers or the racemates of the compounds of formula (I) as MMP inhibitors as well as the use of the (R)-enantiomers as MMP inhibitors is rendered possible by the introduction of the R3 group.
A first aspect of the present invention relates to compounds of formula (I)
wherein
Preferably, in the compounds of formula (I)
The pyrid-3-yl-CH2— or the phenyl-CH2— may be optionally substituted. Preferably, the substituent(s) when present is/are on the ring.
The compounds according to formula (I) have the carbon atom bearing radical R3 with a (S) configuration or the compounds according to formula (I) have the carbon atom bearing radical R3 with a (R) configuration.
In another aspect, the present invention relates to compounds of formula (I) wherein R denotes benzyl, 2-picolyl, and in particular 3-picolyl.
In another aspect, the present invention relates to compounds of formula (I) wherein R1 is —CH3, —CH2—CH2F, —CH2—CH2−18F, CH2—CH2—Cl, —CH2—CH2—OH or —CH2—CH2-OTs.
In another aspect, the present invention relates to compounds of formula (I) wherein R2 is —NH—OH, —O-tert-butyl, or —OCH3.
In another aspect, the present invention relates to compounds of formula (I) wherein R3 is selected from —CH2—CH2F, —CH2—CH2−18F, CH2—C(F)═CH2, —CH2—C(18F)═CH2 and CH2—CH═CH2.
In another aspect, the present invention relates to compounds wherein R4 is selected from H, tert-butyl and methyl.
In another aspect the present invention relates to compounds of formula (I) selected from:
and the corresponding racemates, ester and pharmaceutically acceptable salts thereof.
In another aspect, the present invention relates to compounds of formula (I) which are substituted with 18F. It will be understood that other radionuclides may be used other than 18F. Such radionuclides may include radionuclides useful for PET imaging, which include 11C, 13N, 15O, 18F, 64Cu and 124I, or those useful for SPECT imaging, such as 99Tc, 77Br, 61Cu, 153Gd, 123I, 125I, 131I and 32P.
In another aspect, the present invention relates to compounds of formula (I) wherein R3 is selected from C1-C6(18F) alkyl, C2-C6(18F) alkenyl. More preferably, R3 is CH2CH2—18F or —CH2—C(18F)═CH2.
In another aspect, the present invention relates to compounds of formula (I) wherein R1 is C1-C6 alkyl substituted with a 18F. More preferably, R1 is CH2CH2—18F.
In another aspect, the present invention relates to compounds of formula (I) wherein radical R is substituted with a 18F. Preferably, 18F is on the aryl or heteroaryl moiety of radical R or on a substituent of the aryl or heteroaryl moiety of radical R.
R5 in the compounds of formula (I) may be 18F.
Preferably, the compounds of formula (I) bear one 18F. In this case, 18F may be on a carbon atom of radical R or R1 or R3, preferably as specified above or R5 can be 18F.
The compounds according to formula (I) as described above have the carbon atom bearing radical R3 with a (S) configuration or a (R) configuration.
In another aspect, the present invention relates to compounds of formula (I) for use as a medicament.
In another aspect, the present invention relates to compounds of formula (I) which are labeled with a 18F atom for use as a diagnostic agent, in particular as an in vivo diagnostic agent and more in particular in Positron Emission Tomography (PET).
In a further aspect, the present invention relates to compounds for use in the prevention and/or treatment of pathological conditions associated with unpaired expression of matrix-metalloproteases in human and animal, in particular mammals.
In another aspect, the present invention relates to compounds of formula (I) labeled with a 18F for use in the diagnosis of pathological conditions associated with unpaired expression of metalloproteases in human and animal, in particular mammals. In particular, the compounds are used in the in vivo diagnostic imaging, more in particular in PET.
The pathological conditions are selected from the group consisting of cardiovascular diseases, inflammatory diseases and malignant diseases. More in particular, the cardiovascular diseases are selected from atherosclerosis and congestive heart failure, the inflammatory disease is a chronic obstructive pulmonary disease, the malignant diseases are cancers.
The use of the diagnostic imaging compound of the invention permits the identification of active plaque burden, which allows risk stratification of patients with known or suspected coronary artery disease, i.e. patients with pain or a history of pain, or identified as high risk but asymptomatic. In addition, the diagnostic imaging agents of the invention permit identification of vulnerable plaques in symptomatic patients, which allows identification of high risk of acute myocardial infarction or stroke irrespective of stenosis and permits immediate risk stratification when the patient presents with chest pain. Furthermore, angioplasty of vulnerable plaques is high risk, and may lead to embolism of the artery tree post surgery. Thus imaging of this subtype of plaques may help reduce post-surgical complication.
In a further aspect the present invention relates to pharmaceutical compositions comprising a compound of formula (I).
Suitable preparations include for example tablets, capsules, suppositories, solutions, —particularly solutions for injection (s.c, i.v., i.m.) and infusion—syrups, elixirs, solution for inhalation.
The invention relates to pharmaceutical compositions comprising an effective amount, especially an amount effective in the treatment of one of the above-mentioned disorders, of the active ingredient together with pharmaceutically acceptable carriers that are suitable for topical, enteral, for example oral or rectal, or intravenous or parenteral administration and that may be inorganic or organic, solid or liquid. They are used for oral administration, especially tablets or gelatin capsules that comprise the active ingredient together with diluents, for example lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and/or glycerol, and/or lubricants, for example silica, talc, stearic acid or salts thereof, such as magnesium or calcium stearate, and/or polyethylene glycol. Tablets may also comprise binders, for example magnesium aluminum silicate, starches, such as corn, wheat or rice starch, gelatin, methylcellulose, sodium carboxymethylcellulose and/or polyvinylpyrrolidone, and, if desired, disintegrators, for example starches, agar, alginic acid or a salt thereof, such as sodium alginate, and/or effervescent mixtures, or adsorbents, dyes, flavorings and sweeteners.
It is also possible to use the pharmacologically active compounds of the present invention in the form of intravenously and parentally administrable compositions or in the form of infusion solutions. Such solutions are preferably isotonic aqueous solutions or suspensions which, for example in the case of lyophilized compositions that comprise the active ingredient alone or together with a carrier, for example mannitol, can be made up prior to use. The pharmaceutical compositions may be sterilized and/or may comprise excipients, for example preservatives, stabilizers, wetting agents and/or emulsifiers, solubilizes, salts for regulating the osmotic pressure and/or buffers. The present pharmaceutical compositions are prepared in a manner known per se, for example by means of conventional mixing, granulating, confectioning, dissolving or lyophilizing processes, and comprise approximately from 1% to 95%, especially from approximately 1% to approximately 20%, active ingredient(s).
The present invention further provides a pharmaceutical composition which comprises a compound of formula (I) labeled with a 18F together with a biocompatible carrier, in a form suitable for mammalian, more in particular human administration.
In a further aspect, the present invention relates to a precursor useful in the preparation of the compounds of formula (I) labeled with a 18F atom. The precursor comprises a group suitable for reaction with 18F to give said compound of formula (I). Suitable precursors of the invention for preparation of imaging compounds of formula (I) are compounds of formula (I) which comprise a non-radioactive group (e.g. tosyl group) to permit 18F exchange, preferably via nucleophilic substitution reaction. Preferably, the radiofluorination with 18F-fluoride occurs via nucleophilic substitution reaction of a tosyl group on a precursor compound of the compounds of formula (I) with [18F] (Kryptofix222)KF.
Preferably the precursors are compounds of formula (II)
wherein R and R2 are as defined above in formula (I); R1 is an optionally substituted C1-C6 alkyl (preferably a C2 alkyl), wherein the substituent is a OH, OTs or another leaving group for nucleophilic substitution; R3 is an optionally substituted C1-C6 alkyl or an optionally substituted C2-C6 alkenyl wherein the substituent is a OH, OTs or suitable leaving group for nucleophilic substitution, with the proviso that at least one of R1 and R3 is substituted with a OH, OTs or another leaving group for nucleophilic substitution and/or with the proviso that when R1 is substituted with a OH, OTs or another leaving group for nucleophilic substitution then R3 is C1-C6 fluoro alkyl or C2-C6 alkenyl optionally substituted with F. Preferably, in formula (II): R1 is CH2—CH2-OTs and R3 is CH2—CH2F or CH2—C(F)═CH2 or CH2—C(H)═CH2; or R1 is CH3 and R3 is CH2—CH2OTs or CH2—C(OTs)=CH2.
In another aspect, the invention provides a process for preparing compounds of formula (I) having at least one carbon atom substituted with a 18F. The process comprises reacting a compound of formula (II)
as defined above with a 18F containing reagent for substitution of a OH, OTs or another leaving group with 18F, preferably for substitution of OTs. Preferably, in formula (II):
R1 is CH2—CH2-OTs and R3 is CH2—CH2—F or CH2—C(F)═CH2; or R1 is CH3 and R3 is CH2—CH2-OTs. Preferably, the 18F containing reagent is [18F] (Kryptofix222)KF.
In a further aspect, the present invention relates to compounds of formula (III) that are intermediate compounds in the synthesis of the compounds of the invention:
wherein
Compounds of formula (I) wherein R2 is C1-C6 alkoxy can also be used as intermediate compounds in the synthesis of compounds of formula (I). In particular, they may be used in the synthesis of compounds of formula (I) wherein R2 is NH—OR4 and R4 is H, C1-C6 alkyl, C3-C6 cycloalkyl or C5-C6 heterocycloalkyl.
Compounds of formula (I) wherein R2 is NH—OR4 and R4 is C1-C6 alkyl, C3-C6 cycloalkyl or C5-C6 heterocycloalkyl can also be used as intermediate compound in the synthesis of compounds of formula (I). In particular, they are used in the synthesis of compounds of formula (I) wherein R2 is NH—OR4 and R4 is H.
Intermediates according to the invention are for example compounds having the following structures:
wherein X is selected from F, Cl, OH, OTs and ester thereof.
A further aspect of the present invention is a process for preparing a compound of formula (I)
wherein
wherein
R4O—NH2 formula (IV)
wherein R4 is as defined above.
A compound obtained in step a) having R4 selected from C1-C6 alkyl, C3-C6 cycloalkyl or C5-C6 heterocycloalkyl may be optionally hydrolyzed to a compound wherein R4 is H.
The compound of formula (III′) is prepared by
R—Cl formula (VIII)
Preferably, in step a′) pyridine is used as a solvent. Preferably, the reaction of step a″) is conducted in dimethylformamide (DMF) in the presence of potassium carbonate. Preferably, the reaction of step a) is conducted in the presence of dichloromethane, 1-Hydroxy-benzotriazol (HOB T), N-methyl-morpholin (NMM) and 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide (EDC) hydrochloride.
In a further aspect, the present invention provides a kit for the preparation of the pharmaceutical composition of the invention. Where the pharmaceutical composition of the invention comprises a diagnostic imaging agent labeled with a 18F, said kit comprises a precursor which is a compound of formula (I) comprising a group (e.g. tosyl group) suitable for reaction with a 18F containing compound (e.g. [18F] (Kryptofix222)KF) such that reaction of said precursor with said 18F containing compound gives said diagnostic imaging agent product of formula (I). Preferably said precursor is a compound of formula (II).
The kits may optionally further comprise additional components such as a radioprotectant, antimicrobial preservative, pH-adjusting agent or filler.
The term “C1-C6 alkyl”, when used either alone or within other terms such as “haloalkyl” and “arylalkyl” or “heteroarylalkyl” embraces linear or branched radicals having C1-C6 carbon atoms. Examples of such radicals include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl, isoamyl, hexyl. The term “alkylen” embraces bridging divalent alkyl radicals such as methylen and ethylen.
The term “C2-C6 alkenyl”, when used alone or in combination, embraces linear or branched radicals having at least one carbon-carbon double bond in a moiety having between two and six carbon atoms. Examples of alkenyl radicals include, without limitation, vinyl, propenyl, allyl, butenyl and 4-methylbutenyl. The term “alkenyl” encompasses radicals having “cis” and “trans” orientations, or alternatively, “E” and “Z” orientations, as appreciated by those of ordinary skill in the art.
The term “C1-C6 alkoxy” when used alone or in combination, embraces linear or branched oxygen-containing radicals each having alkyl portions of one to six carbon atoms. Examples of such radicals include methoxy, ethoxy, propoxy, butoxy and tert-butoxy. Alkoxy radicals may be further substituted with one or more halo atoms, such as fluoro, chloro or bromo, to provide “haloalkoxy” radicals. Examples of such radicals include fluoromethoxy, chloromethoxy, trifluoromethoxy, trifluoroethoxy, fluoroethoxy, fluoropropoxy and fluorobutoxy.
The term “halo”, when used alone or in combination, means halogens such as fluorine, chlorine, bromine or iodine atoms, preferably fluorine. The term “haloalkyl”, when used alone or in combination, embraces radicals wherein any one or more of the alkyl carbon atoms is substituted with halo as defined above. For example, this term includes monohaloalkyl, dihaloalkyl and polyhaloalkyl radicals such as a perhaloalkyl. A monohaloalkyl radical, for example, may have either an iodo, bromo, chloro or fluoro atom within the radical, preferably a fluoro atom. Dihalo and polyhaloalkyl radicals may have two or more of the same halo atoms or a combination of different halo radicals.
The term “C1-C6 haloalkyl” embraces radicals having 1-6 carbon atoms and, for example, haloalkyl radicals having one to three carbon atoms. Examples of haloalkyl radicals include fluoromethyl, difluoromethyl, trifluoromethyl, chloromethyl, dichloromethyl, trichloromethyl, pentafluoroethyl, heptafluoropropyl, chlorodifluoromethyl, dichlorofluoromethyl, fluoroethyl, difluoroethyl, trifluoroethyl, fluoropropyl, difluoropropyl, trifluoropropyl, chloroethyl, dichloroethyl and dichloropropyl.
The term “C2-C6 haloalkenyl”, when used alone or in combination, embraces radicals wherein anyone or more of the alkyl carbon atoms is substituted with halo as defined above, preferably a fluorine atom. For example, this term includes monohaloalkenyl, dihaloalkenyl and polyhaloalkenyl radicals. A monohaloalkenyl radical, for example, may have either an iodo, bromo, chloro or fluoro atom within the radical. Dihalo and polyhaloalkenyl radicals may have two or more of the same halo atoms or a combination of different halo radicals. Examples of haloalkenyl radicals include fluorovinyl, fluoropropenyl and fluorobutenyl.
The term “aryl”, when used alone or in combination, means a carbocyclic aromatic moiety containing one, two or even three rings wherein such rings may be attached together in a fused manner. Thus, the term “aryl” embraces aromatic radicals such as phenyl, naphthyl, indenyl, tetrahydronaphthyl, dihydrobenzofuranyl, anthracenyl, indanyl, benzodioxazinyl. The “aryl” group may be substituted, such as with 1 to 5 substituents including C1-C6 alkyl, hydroxyl, halo, halo-C1-C6-alkyl, nitro, cyano, C1-C6 alkoxy and C1-C6 alkylamino.
The term “heteroaryl”, as used herein, either alone or in combination, means a fully unsaturated (aromatic) ring moiety formed from carbon atoms and having one or more heteroatoms selected from nitrogen, oxygen and sulfur. The ring moiety or ring system may contain one (“monocyclic”), two (“bicyclic”) or even three (“tricyclic”) rings wherein such rings are attached together in a fused manner. Every ring of a “heteroaryl” ring system need not be aromatic, and the ring(s) fused thereto (to the heteroaromatic ring) may be partially or fully saturated and optionally include one or more heteroatoms selected from nitrogen, oxygen and sulfur.
Examples of unsaturated heteroaryl radicals, include unsaturated 5- to 6-membered heteromonocyclyl groups containing 1 to 4 nitrogen atoms, including for example, pyrrolyl, imidazolyl, pyrazolyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, pyrimidyl, pyrazinyl, pyridazinyl and tetrazole; unsaturated 7- to 10-membered heterobicyclyl groups containing 1 to 4 nitrogen atoms, including for example, quinolinyl, isoquinolinyl, quinazolinyl, isoquinazolinyl, aza-quinazolinyl; unsaturated 5- to 6-membered heteromonocyclic groups containing an oxygen atom, for example, pyranyl, 2-furyl, 3-furyl, benzofuryl, etc.; unsaturated 5 to 6-membered heteromonocyclic groups containing a sulfur atom, for example, 2-thienyl, 3-thienyl, benzothienyl, etc.; unsaturated 5- to 6-membered heteromonocyclic groups containing 1 to 2 oxygen atoms and 1 to 3 nitrogen atoms, for example, oxazolyl, isoxazolyl, oxadiazolyl [e.g., 1,2,4-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,5-oxadiazolyl]; unsaturated 5 to 6-membered heteromonocyclic group containing 1 to 2 sulfur atoms and 1 to 3 nitrogen atoms, for example, thiazolyl, isothiazolyl, thiadiazolyl [e.g. 1,2,4-thiadiazolyl, 1,3,4-thiadiazolyl, 1,2,5-thiadiazolyl].
The term “aryl-alkyl” is any C7-C20 group consisting of an alkyl and an aryl group as defined above, preferably a benzyl group. Preferably the alkyl moiety of the “arylalkyl” group is a —CH2- or a —CH2—CH2— group. Preferably the aryl moiety of the “arylalkyl” group is a phenyl. The “arylalkyl” may be optionally substituted with one or more substituents independently selected from OH, OTs, amino, halo in particular F and C1, C1-C6 alkyl optionally substituted with OH, OTs, amino, halo in particular F and Cl or the “arylalkyl” may be optionally substituted with —O(CH2—CH2—O)nCH2—CH2—R5 wherein n is 1, 2, 3, 4, 5, 6, preferably 3, and R5 can be F, 18F, Cl, OH, OTs; or —(CH2—CH2—O)nCH2—CH2—R5 wherein n is 1, 2, 3, 4, 5, 6, preferably 3, and R5 can be F, 18F, Cl, OH, OTs. Preferably the substituent(s) are on the “aryl” moiety.
The term “heteroaryl-alkyl” is any C7-C20 group consisting of an alkyl and a heteroaryl group as defined above, preferably a picolyl group, more preferably a 3-picolyl group. Preferably, the alkyl moiety of the “heteroaryl-alkyl” group is a —CH2— or a —CH2—CH2— group. Preferably, the heteroaryl moiety of the “heteroaryl-alkyl” group is a pyridyl, more preferably a 3-pyridyl. The “heteroaryl-alkyl” may be optionally substituted with one or more substituents independently selected from OH, OTs, amino, halo in particular F and C1, C1-C6 alkyl optionally substituted with OH, OTs, amino, halo in particular F and Cl; the “heteroaryl-alkyl” may be optionally substituted with —O(CH2—CH2—O)nCH2—CH2—R5 wherein n is 1, 2, 3, 4, 5, 6, preferably 3, and R5 can be F, 18F, Cl, OH, OTs; or —(CH2—CH2—O)nCH2—CH2—R5 wherein n is 1, 2, 3, 4, 5, 6, preferably 3, and R5 can be F, 18F, Cl, OH, OTs. Preferably the substituent(s) are on the “aryl” moiety.
The term “C5-C6-heterocycloalkyl” relates to saturated or unsaturated mono ring comprising 5 or 6 carbon atoms, which carry heteroatoms such as N, O, S instead of one or more carbon atoms. Examples of such heterocyclic group are tetrahydrofuranyl, tetrahydropyranyl, pyrrolidinyl, pyrrolidino, piperidinyl, piperidino, piperazinyl, piperazino, morpholinyl, morpholino, thiomorpholinyl, thiomorpholino, tetrahydrothiofuranyl, and pyranyl
“Leavings group for nucleophilic substitution” are known to the skilled person in the art. They include e.g., Cl, Br, I etc.
The phrase “labeled with a 18F” used herein means that a compounds of the invention comprises a 18F either as an artificially enriched level of an atom intrinsic to the substructure, or as an additional essential feature that has been chemically attached via a functionality suitable for coupling said 18F.
By the term “biocompatible carrier” is meant a fluid, especially a liquid, in which the imaging agent can be suspended or dissolved, such that the composition is physiologically tolerable, i.e. it can be administered to the mammalian body without toxicity or undue discomfort. The biocompatible carrier is suitably an injectable carrier liquid such as sterile, pyrogen-free water for injection; an aqueous solution such as saline (which may advantageously be balanced so that the final product for injection is either isotonic or not hypotonic); an aqueous solution of one or more tonicity-adjusting substances (e.g. salts of plasma cations with biocompatible counter-ions), sugars (e.g. glucose or sucrose), sugar alcohols (e.g. sorbitol or mannitol), glycols (e.g. glycerol), or other non-ionic polyol materials (e.g. polyethyleneglycols, propylene glycols and the like).
By the term “biocompatible cation” is meant a positively charged counter-ion which forms a salt with an ionized, negatively charged group, where said positively charged counter-ion is also non-toxic and hence suitable for administration to the mammalian body, especially the human body. Examples of suitable biocompatible cations include the alkali metals sodium or potassium; the alkaline earth metals calcium and magnesium; and the ammonium ion. Preferred biocompatible cations are sodium and potassium, most preferably sodium.
By the term “radioprotectant” is meant a compound which inhibits degradation reactions, such as redox processes, by trapping highly-reactive free radicals, such as oxygen-containing free radicals arising from the radiolysis of water. The radioprotectants of the present invention are suitably chosen from: ascorbic acid, para-aminobenzoic acid (i.e. 4-aminobenzoic acid), gentisic acid (i.e. 2,5-dihydroxybenzoic acid) and salts thereof with a biocompatible cation as described above.
By the term “antimicrobial preservative” is meant an agent which inhibits the growth of potentially harmful micro-organisms such as bacteria, yeasts or moulds. The antimicrobial preservative may also exhibit some bactericidal properties, depending on the dose. The main role of the antimicrobial preservative(s) of the present invention is to inhibit the growth of any such micro-organism in the pharmaceutical composition post-reconstitution, i.e. in the radioactive diagnostic product itself. The antimicrobial preservative may, however, also optionally be used to inhibit the growth of potentially harmful microorganisms in one or more components of the kit of the present invention prior to reconstitution. Suitable antimicrobial preservatives include: the parabens, i.e. methyl, ethyl, propyl or butyl paraben or mixtures thereof; benzyl alcohol; phenol; cresol; cetrimide and thiomersal. Preferred antimicrobial preservative(s) are the parabens.
The term “pH-adjusting agent” means a compound or mixture of compounds useful to ensure that the pH of the reconstituted kit is within acceptable limits (approximately pH 4.0 to 10.5) for human or mammalian administration. Suitable such pH-adjusting agents include pharmaceutically acceptable buffers, such as tricine, phosphate or TRIS [i.e. tris(hydroxymethyl)aminomethane], and pharmaceutical acceptable bases such as sodium carbonate, sodium bicarbonate or mixtures thereof. When the ligand conjugate is employed in acid salt form, the pH-adjusting agent may optionally be provided in a separate vial or container, so that the user of the kit can adjust the pH as part of a multi-step procedure.
By the term “filler” is meant a pharmaceutically acceptable bulking agent which may facilitate material handling during production and lyophilisation. Suitable fillers include inorganic salts such as sodium chloride, and water soluble sugars or sugar alcohols such as sucrose, maltose, mannitol or trehalose.
a illustrates the synthetic route that was used to prepare compounds 1a, 1b, 3a, 3b, 4a, 4b, 5a and 5b, respectively.
b illustrates the synthetic route that was used to prepare compounds 6a, 6b, 8a, 8b, 9a, 9b, 10a and 10b, respectively.
c illustrates the synthetic route that was used to prepare compounds 16a, 16b, 17a, 17b, 18a, 18b, 19a and 19b, respectively.
a-c illustrate the synthetic route that was used to prepare the compounds of formula (I) wherein R1 is F—CH2—CH2, TsO—CH2—CH2, HO—CH2—CH2, Cl—CH2—CH2, respectively.
The syntheses disclosed in
According to the general procedure tert-butyl (S)-aminobutanoate (630 mg, 3.96 mmol) in pyridin (15 mL) was treated with p-methoxyphenylsulfonyl chloride (818 mg, 3.96 mmol). The crude product was purified by column chromatography to get a white solid. Yield: 467 mg (36%). M.p. 82° C. 1H NMR (300 MHz, CDCl3): δ 0.92 (t, 3JH,H=7.4 Hz, 3H, 4-CH3), 1.27 (s, 9H, 6-CH3), 1.61 (m, 1H, 3-CHA), 1.77 (m, 1H, 3-CHB), 3.72 (ddd, 3JH,H=9.1 Hz, 3JH,H=6.9 Hz, 3JH,H=5.2 Hz, 1H, 2-CH), 3.85 (s, 3H, 11-CH3), 5.19 (d, 3JH,H=9.1 Hz, 1H, 12-NH), 6.95 (dm, 3JH,H=9.0 Hz, 2H, 8-CH), 7.78 (dm, 3JH,H=9.0 Hz, 2H, 9-CH). 13C NMR (75 MHz, CDCl3): δ 9.2 (q, C-4), 26.8 (q, C-3), 27.7 (t, C-6), 55.6 (q, C-11), 57.1 (d, C-2), 82.2 (s, C-5), 114.1 (d, C-8), 129.4 (d, C-9), 131.4 (s, C-7), 162.9 (s, C-10), 170.8 (s, C-1). Exact mass (ESI+): C15H23NO5S+Na+, calcd. 352.1195. found 352.1192, (C15H23NO5S)2+Na+, calcd. 681.2492. found 681.2483. MS (GC/MS, 70 eV): m/z (%)=329 (1) [M+], 228 (88) [M+—C5H9O2], 171 (100) [M+—C7H7O3S], 155 (5) [C6H5NO2S+], 123 (18), 107 (21) [C7H7O+], 77 (13) [C6H5+], 57 (15) [C4H9+]. Optical rotation: [α]58920=+21.3, [α]57820=22.3, [α]54620=26.2, [α]43620=54.4, [α]36520=110.5 (c=1.009, CHCl3).
According to the general procedure tert-butyl (R)-aminobutanoic acid (551 mg, 3.46 mmol) was dissolved in pyridin (15 mL) and treated with p-methoxyphenylsulfonyl chloride (715 mg, 3.46 mmol). The crude product was purified by column chromatography to get a white solid. Yield: 446 mg (39%). M.p. 81° C. 1H NMR (400 MHz, CDCl3): δ 0.92 (t, 3JH,H=7.4 Hz, 3H, 4-CH3), 1.27 (s, 9H, 6-CH3), 1.64 (m, 1H, 3-CHA), 1.76 (ddd, 2JH,H=14.7 Hz, 3JH,H=7.4 Hz, 3JH,H=5.2 Hz, 1H, 3-CHB), 3.72 (ddd, 3JH,H=9.1 Hz, 3JH,H=7.0 Hz, 3JH,H=5.2 Hz, 1H, 2-CH), 3.85 (s, 3H, 11-CH3), 5.18 (d, 3JH,H=9.1 Hz, 1H, 12-NH), 6.95 (dm, 3JH,H=9.0 Hz, 2H, 8-CH), 7.78 (dm, 3JH,H=9.0 Hz, 2H, 9-CH3). 13C NMR (100 MHz, CDCl3): δ 9.2 (q, C-4), 26.8 (t, C-3), 27.7 (q, C-6), 55.6 (q, C-11), 57.1 (d, C-2), 82.3 (s, C-5), 114.1 (d, C-8), 129.4 (d, C-9), 131.4 (s, C-7), 162.9 (s, C-10), 170.8 (s, C-1). Exact mass (ESI+): C15H23NO5S+Na+, calcd. 352.1195. found 352.1189; (C15H23NO5S)2+Na+, calcd. 681.2492. found 681.2483. MS (GC/MS, 70 eV): m/z (%), 29 (3) [M+], 273 (1) [M+—C4H8], 228 (100) [M+—C5H9O2], 171 (90) [C7H7O3S+], 155 (4) [C6H5NO2S+], 123 (22), 107 (29) [C7H7O+], 92 (10) [C6H4O+], 77 (15) [C6H5+], 57 (15) [C4H9+], 56 (48) [C4H8+]. Optical rotation: [α]58920=−20.6, [α]57820=−22.0, [α]54620=−26.8, [α]43620=−54.1, [α]36520=−109.9 (c=1.008, CHCl3).
According to the general procedure 558 mg (3.63 mmol) methyl (S)-aminobutanoate hydrochloride was dissolved in pyridin (30 mL) and treated with p-methoxyphenylsulfonyl chloride (751 mg, 3.63 mmol). The crude product was purified by column chromatography to get a white solid. Yield: 450 mg (43%). M.p. 64° C. 1H NMR (300 MHz, CDCl3): δ 0.91 (t, 3JH,H=7.4 Hz, 3H, 4-CH3), 1.60-1.86 (m, 2H, 3-CH2), 3.52 (s, 3H, 5-CH3), 3.86 (s, 3H, 10-CH3), 3.85 (m 1H, 2-CH), 5.30 (d, 3JH,H=9.2 Hz, 1H, 11-NH), 6.93-6.99 (m, 2H, 7-CH), 7.75-7.81 (m, 2H, 8-CH). 13C NMR (75 MHz, CDCl3): δ 9.3 (q, C-4), 26.6 (t, C-3), 52.4 (q, C-5), 55.6 (q, C-10), 56.7 (d, C-2), 114.0 (d, C-7), 129.3 (d, C-8), 131.2 (s, C-6), 162.9 (s, C-9), 172.1 (s, C-1). Elemental analysis: C12H17NO5S (M=287.332 g/mol), calcd. C, 50.16; H, 5.96; N, 4.87. found C, 50.16; H, 5.91; N, 4.91%. Exact mass (ESI+): C12H17NO5S+H+, calcd. 288.0906. found 288.0891. C12H17NO5S+Na+, calcd. 310.0725. found 310.0718; (C12H17NO5S)2+Na+, 597.1553. found 597.1546. MS (GC/MS, 70 eV): m/z (%) 287 (5) [M+], 228 (70) [M+—CO2CH3], 171 (100) [C7H7O3S+], 155 (5) [C7H7O2S+], 123 (23) [C7H7O2+], 107 (38) [C7H7O+], 92 (9), 77 (12) [C6H5+], 64 (6) [SO2+], 59 (3) [CO2CH3+]. Optical Rotation: [α]58920=+8.4, [α]57820=+8.7, [α]54620=+10.7, [α]43620=+27.0, [α]36520=+64.0 (c=1.001, CHCl3).
According to the general procedure methyl (R)-aminobutanoate hydrochloride (1000 mg, 6.51 mmol) was dissolved in pyridin (30 mL) and treated with p-methoxyphenylsulfonyl chloride (1350 mg, 6.51 mmol). The crude product was purified by column chromatography to get a white solid. Yield: 879 mg (47%). M.p. 63-64° C. 1H NMR (300 MHz, CDCl3): δ 0.91 (t, 3JH,H=7.4 Hz, 3H, 4-CH3), 1.60-1.84 (m, 2H, 3-CH2), 3.52 (s, 3H, 5-CH3), 3.86 (s, 3H, 10-CH3), 3.85 (m 1H, 2-CH), 5.24 (d, 3JH,H=9.2 Hz, 1H, 11-NH), 6.92-6.99 (m, 2H, 7-CH), 7.73-7.83 (m, 2H, 8-CH). 13C NMR (75 MHz, CDCl3): δ 9.3 (q, C-4), 26.6 (t, C-3), 52.4 (q, C-5), 55.6 (q, C-10), 56.7 (d, C-2), 114.1 (d, C-7), 129.3 (d, C-8), 131.2 (s, C-6), 162.9 (s, C-9), 172.1 (s, C-1). Exact mass (ESI+): C12H17NO5S+H+, calcd. 288.0906. found 288.0913; C12H17NO5S+Na+, calcd. 310.0725. found 310.0733; (C12H17NO5S)2+Na+, calcd. 597.1553. found 597.1550. MS (ESI+, daughter ion experiment): m/z (%)=288 (9) [M++H+], 228 (100) [M+−C2H3O2], 171 (32) [C7H7O3S+].
According to the general procedure tert-butyl (S)-aminopent-4-enoate (1200 mg, 7.02 mmol) was dissolved in pyridin (20 mL) and treated with p-methoxyphenylsulfonyl chloride (1454 mg, 7.02 mmol). The crude product was purified by column chromatography to get a white solid. Yield: 1150 mg (48%). M.p. 58-59° C. 1H NMR (300 MHz, CDCl3): δ 1.29 (s, 9H, 7-CH3), 2.40-2.48 (m, 2H, 3-CH2), 3.85 (s, 3H, 12-CH3), 3.88 (dt, 3JH,H=9.0 Hz, 3JH,H=5.8 Hz, 1H, 13-NH), 5.05-5.13 (m, 2H, 5-CH2), 5.20 (d, 3JH,H=8.9 Hz, 1H, 2-CH), 5.66 (ddt, 3JH,H=15.9 Hz, 3JH,H=11.2 Hz, 3JH,H=7.1 Hz, 1H, 4-CH), 6.95 (dm, 3JH,H=9.0 Hz, 2H, 9-CH), 7.78 (dm, 3JH,H=9.0 Hz, 2H, 10-CH). 13C NMR (75 MHz, CDCl3): δ 27.8 (q, C-7), 37.9 (t, C-3), 55.5 (d, C-2), 55.6 (q, C-12), 82.6 (s, C-6), 114.2 (d, C-9), 119.3 (t, C-5), 129.4 (d, C-10), 131.6 (d, C-4 and C-8), 163.0 (s, C-11), 170.0 (s, C-1).
According to the general procedure tert-butyl (R)-aminopent-4-enoate (2140 mg, 12.48 mmol) was dissolved in pyridin (25 mL) and treated with p-methoxyphenylsulfonyl chloride (2570 mg, 12.48 mmol). The crude product was purified by column chromatography to get a white solid. Yield: 2375 mg (56%). M.p. 59° C. 1H NMR (300 MHz, CDCl3): δ 1.27 (s, 9H, 7-CH3), 2.39-2.50 (m, 2H, 3-CH2), 3.85 (s, 3H, 12-CH3), 3.88 (dt, 3JH,H=8.6 Hz, 3JH,H=5.5 Hz, 1H, 13-NH), 5.04-5.13 (m, 2H, 5-CH2), 5.20 (d, 3JH,H=9.0 Hz, 1H, 2-CH), 5.67 (m, 1H, 4-CH), 6.95 (dm, 3JH,H=9.0 Hz, 2H, 9-CH), 7.78 (dm, 3JH,H=9.0 Hz, 2H, 10-CH). 13C NMR (75 MHz, CDCl3): δ 27.7 (q, C-7), 37.9 (t, C-3), 55.4 (d, C-2), 55.6 (q, C-12), 82.5 (s, C-6), 114.1 (d, C-9), 119.4 (t, C-5), 129.4 (d, C-10), 131.4 (s, C-8), 131.5 (d, C-4), 162.9 (s, C-11), 169.9 (s, C-1). Exact mass (ESI+): C16H23NO5S+Na+, calcd. 364.1195. found 364.1187; (C16H23NO5S)2+Na+, calcd. 705.2492. found 705.2488. MS (GC/MS, 70 eV): m/z (%) 364 (0) [M+], 300 (10) [M+-C3H5], 285 (4) [M+—C4H8], 240 (52) [M+—C5H9O2], 171 (100) [C7H7O3S+], 155 (5) [C6H5NO2S+], 123 (36), 107 (50) [C7H7O+], 92 (13) [C6H4O+], 77 (24) [C6H5+], 64 (8) [SO2+], 57 (42) [C4H9+], 56 (48) [C4H8+], 41 (40) [C3H5+]. Optical rotation: [α]58920=−17.1, [α]57820=−18.4, [α]54620=−20.4, [α]43620=−43.8, [α]36520=−88.5 (c=1.010, CHCl3).
According to the general procedure tert-butyl (S)-2-amino-4-fluorobutanoate (1600 mg, 9.03 mmol) was dissolved in pyridin (15 mL) and treated with p-methoxyphenylsulfonyl chloride (1860 mg, 9.03 mmol). The crude product was purified by column chromatography to get a white solid. Yield: 2200 mg (70%). M.p. 89-90° C. 1H NMR (300 MHz, CDCl3): δ 1.28 (s, 9 H, 6-CH3), 1.87-2.25 (m, 3JH,F=26.2 Hz, 3JH,H=5.5 Hz, 2H, 3-CH2), 3.85 (s, 3H, 11-CH3), 3.89 (m, 3JH,H=8.0 Hz, 1H, 2-CH), 4.58 (dt, 2JH,F=46.8 Hz, 3JH,H=5.5 Hz, 2H, 4-CH2F), 5.34 (d, 3JH,H=8.0 Hz, 1H, 12-NH), 6.96 (dm, 3JH,H=8.8 Hz, 2H, 8-CH), 7.79 (dm, 3JH,H=8.8 Hz, 2H, 9-CH). 13C NMR (75 MHz, CDCl3): δ 27.6 (q, C-6), 34.1 (dt, 2JH,F=20.3 Hz, C-3), 52.7 (dd, 3JH,F=3.7 Hz, C-2), 55.6 (q, C-11), 79.7 (dt, 1JH,F=165.9 Hz, C-4), 82.9 (s, C-5), 114.2 (d, C-8), 129.5 (d, C-9), 130.9 (s, C-7), 163.1 (s, C-10), 170.2 (s, C-1). 19F NMR (282 MHz, CDCl3): δ −221.6 (tt, 2JH,F=46.8 Hz, 3JH,F=26.0 Hz, 4-CH2F). Elemental analysis: C15H22FNO5S (M=347.402 g/mol), calcd. C, 51.86; H, 6.38; N, 4.03. found C, 51.92; H, 6.24; N, 3.96%. Exact mass (ESI+): C15H22FNO5S+Na+, cacld. 370.1100. found 370.1094. (C15H22FNO5S)2+Na+, calcd. 717.2303. found 717.2298. Optical rotation: [α]58920=−23.5, [α]57820=−25.1, [α]54620=−29.3, [α]43620=−58.4, [α]36520=−114.1 (c=1.021, CHCl3).
According to the general procedure tert-butyl (R)-2-amino-4-fluorobutanoate (1480 mg, 8.35 mmol) was dissolved in pyridin (15 mL) and treated with p-methoxyphenylsulfonyl chloride (1730 mg, 8.35 mmol). The crude product was purified by column chromatography to get a white solid. Yield: 1190 mg (41%). M.p. 89° C. 1H NMR (400 MHz, CDCl3): δ 1.28 (s, 9H, 6-CH3), 1.90-2.23 (m, 2H, 3-CH2), 3.85 (s, 3H, 11-CH3), 3.89 (m, 1H, 12-NH), 4.58 (dt, 2JH,F=46.8 Hz, 3JH,H=5.5 Hz, 2H, 4-CH2F), 5.38 (d, 3JH,H=8.6, 1 H, 2-CH), 6.87-7.07 (m, 2H, 8-CH), 7.75-7.82 (m, 2H, 9-CH). 13C NMR (100 MHz, CDCl3): δ 27.6 (q, C-6), 34.1 (dt, 2JC,F=20.3 Hz, C-3), 52.7 (dt, 3JC,F=3.7 Hz, C-2), 55.6 (q, C-11), 79.7 (dt, 1JH,F=165.9 Hz, C-4), 82.9 (s, C-5), 114.2 (s, C-8), 129.4 (d, C-9), 130.9 (s, C-7), 163.0 (s, C-10), 170.2 (s, C-1). 19F NMR (282 MHz, CDCl3): δ −221.6 (tt, 2JH,F=46.8 Hz, 3JH,F=26.0 Hz, 4-CH2F). Exact mass (ESI+): C15H22FNO5S+Na+, calcd. 370.1100. found 370.1094. MS (GC/MS, 70 eV): m/z (%) 347 (2) [M+], 246 (82) [M+—CO2C(CH3)3], 171 (100) [C7H7O3S+], 155 (5) [177-CH3], 123 (19) [C6H4OS+], 107 (38) [C7H7O+], 92 (18), 77 (27) [C6H5+], 64 (10) [SO2+], 57 (52) [C4H9+]41 (35). Optical rotation: [α]58920=+24.7, [α]57820=+26.1, [α]54620=+30.4, [α]43620=+61.2, [α]36520=+118.4 (c=1.015, CHCl3).
According to the general procedure tert-butyl (S)-2-amino-4-fluoropent-4-enoate (1360 mg, 7.19 mmol) was dissolved in pyridin (20 mL) and treated with p-methoxyphenylsulfonyl chloride (1490 mg, 7.19 mmol). The crude product was purified by column chromatography to get a greenish viscose liquid. Yield: 1.27 g (49%). 1H NMR (300 MHz, CDCl3): δ 1.30 (s, 9H, 7-CH3), 2.58 (dd, 3JH,H=5.7 Hz, 3JH,H=3.6 Hz, 1H, 3-CHA), 2.64 (d, 3JH,H=5.9 Hz, 1 H, 3-CHB), 3.85 (s, 3H, 12-CH3), 3.99 (t, 3JH,H=5.6 Hz, 1H, 2-CH), 4.34 (dd, 3JH,F=49.3 Hz, 2JH,H=3.0 Hz, 1H, 5-CHtrans 4.63 (dd, 3JH,F=17.1 Hz, 2JH,H=3.0 Hz, 1H, 5-CHcis), 5.41 (s, 1H, 13-NH), 6.96 (dm, 3JH,H=9.0 Hz, 2H, 9-CH), 7.79 (dm, 3JH,H=9.0 Hz, 2H, 10-CH). 13C NMR (75 MHz, CDCl3): δ 27.6 (q, C-7), 36.4 (dt, 2JH,F=27.5 Hz, C-3), 53.3 (d, C-2), 55.5 (q, C-12), 83.0 (s, C-6), 93.9 (dt, 2JH,F=18.7 Hz, C-5), 114.1 (d, C-9), 129.4 (d, C-10), 131.3 (s, C-8), 161.0 (ds, 1JH,F=257.1 Hz, C-4), 163.0 (s, C-11), 169.2 (s, C-1). 19F NMR (282 MHz, CDCl3): δ −96.0 (ddt, 3JH,F=49.5 Hz, 3JH,F=19.5 Hz, 3JH,F=17.5 Hz, 1H). MS (ESI+, daughter ion experiment): m/z (%) 360 (0) [M++H], 304 (8) [M+-C4H8], 284 (20) [304-F], 244 (10) [304-C3H4F], 214 (10) [M+-C5H9O2—C2H2F], 188 (15) [M+-C7H7O3S], 171 (100) [C7H7O3S+], 107 (1) [C7H7O+], 88 (60), 77 (2) [C6H5+]. Exact mass (ESI+): C16H22FNO5S+Na+, calcd. 382.1100. found 382.1096. Optical rotation: [α]58920=+3.0, [α]57820=+3.1, [α]54620=+4.1, [α]43620=+13.2, [α]36520=+36.8 (c=1.005, CHCl3).
According to the general procedure tert-butyl (R)-2-amino-4-fluoro-pent-4-enoate (1362 mg, 7.20 mmol) was dissolved in pyridin (20 mL) and treated with p-methoxyphenylsulfonyl chloride (1490 mg, 7.20 mmol). The crude product was purified by column chromatography to get a greenish viscose liquid. Yield: 1.01 g (39%). 1H NMR (400 MHz, CDCl3): δ 1.30 (s, 9H, 7-CH3), 2.45-2.69 (m, 3JH,F=20.5 Hz, 2H, 3-CH2), 3.85 (s, 3H, 12-CH3), 3.98 (dt, 3JH,H=8.9 Hz, 3JH,H=5.8 Hz, 1H, 2-CH), 4.35 (dd, 3JH,F=49.3, 2JH,H=3.0 Hz, 1H, 5-CHtrans), 4.64 (dd, 3JH,F=17.1 Hz, 2JH,H=3.0 Hz, 1H, 5-CHcis), 5.33 (d, 3JH,H=8.9 Hz, 1H, 13-NH), 6.96 (dm, 3JH,H=9.0 Hz, 2H, 9-CH), 7.79 (dm, 3JH,H=9.0 Hz, 2H, 10-CH). 13C NMR (100 MHz, CDCl3): δ 27.6 (q, C-7), 36.5 (dt, 2JH,F=27.5 Hz, C-3), 53.2 (d, C-2), 55.6 (q, C-12), 83.1 (s, C-6), 94.1 (dt, 2JH,F=18.7 Hz, C-5), 114.2 (d, C-9), 129.4 (d, C-10), 131.2 (s, C-8), 161.0 (ds, 1JH,F=257.1 Hz, C-4), 163.0 (s, C-11), 169.2 (s, C-1). 19F NMR (282 MHz, CDCl3): δ −96.1 (ddt, 3JH,F=49.3 Hz, 3JH,F=20.1 Hz, 3JH,F=17.2 Hz, 4-CF). Elemental analysis: C16H22FNO5S (M=359.413 g/mol), calcd. C, 53.47; H, 6.17; N, 3.90. found C, 53.44; H, 6.01; N, 3.83%. MS (GC/MS, 70 eV): m/z (%) 359 (0) [M+], 303 (7) [M+-C4H8], 300 (5) [M+—C3H4F], 258 (52) [M+-CO2C(CH3)3], 244 (21) [258-CH2], 187 (1) [C7H7O3SNH2+], 173 (9) [M+-C7H8O3SNH/C9H14FO2+], 171 (100) [C7H7O3S+], 155 (8) [C6H5NO2S+/C9H15O2+], 123 (21), 107 (28) [C7H7O+], 92 (11), 78 (3) [NO2S+], 77 (19) [C6H5+], 64 (7) [SO2+], 59 (4) [C3H4F+], 57 (24) [C4H9+], 41 (6). Optical rotation: [α]58920=−0.8, [α]57820=−1.0, [α]54620=−1.2, [α]43620=−3.9, [α]36520=−10.2 (c=0.998, CHCl3).
According to the general procedure tert-butyl (S)—N-(4-methoxyphenyl-sulfonyl)amino butanoate (450 mg, 1.37 mmol) was treated with potassium carbonate (1890 mg, 13.70 mmol) and picolyl chloride (225 mg, 1.37 mmol) in DMF (30 mL). After column chromatography (column Ø 3 cm×16 cm) the product was isolated as a white solid. Yield: 505 mg (88%). M.p. 170° C. 1H NMR (400 MHz, CDCl3): δ 0.80 (t, 3JH,H=7.4 Hz, 3H, 4-CH3), 1.31 (s, 9 H, 6-CH3), 1.49 (ddd, 2JH,H=14.1 Hz, 3JH,H=11.5 Hz, 3JH,H=6.1 Hz, 1H, 3-CHA), 1.77 (m, 1H, 3-CHB), 3.84 (s, 3H, 11-CH3), 4.31 (dd, 3JH,H=8.4 Hz, 3JH,H=6.7 Hz, 1H, 2-CH), 4.50 (d, 2JH,H=16.5 Hz, 1H, 12-CHA), 4.71 (d, 2JH,H=16.5 Hz, 1H, 12-CHB), 6.94 (dm, 3JH,H=9.0 Hz, 2H, 8-CH), 7.24 (dd, 3JH,H=7.8 Hz, 3JH,H=4.8 Hz, 1H, 16-CH), 7.72 (dm, 3JH,H=8.9 Hz, 2H, 9-CH), 7.88 (dd, 3JH,H=7.9 Hz, 4JH,H=1.7 Hz, 1H, 17-CH), 8.49 (dd, 3JH,H=4.8 Hz, 4JH,H=1.5 Hz, 1H, 15-CH), 8.56 (d, 4JH,H=1.9 Hz, 1H, 14-CH). 13C NMR (100 MHz, CDCl3): δ 10.7 (q, C-4), 24.2 (t, C-3), 27.6 (q, C-6), 46.2 (t, C-12), 55.4 (q, C-11), 61.9 (d, C-2), 81.7 (s, C-5), 113.9 (d, C-8), 123.0 (d, C-16), 129.3 (d, C-9), 131.4 (s, C-7), 133.7 (s, C-13), 136.0 (d, C-17), 148.5 (d, C-15), 149.2 (d, C-14), 162.7 (s, C-10), 169.7 (s, C-1).
Optical rotation: [α]58920=−43.3, [α]57820=−45.5, [α]54620=−52.5, [α]43620=−96.2, [α]36520=−169.7 (c=1.013, CHCl3):
According to the general procedure tert-butyl (R)—N-(4-methoxyphenylsulfonyl)amino butanoate (338 mg, 1.03 mmol), potassium carbonate (1420 mg, 10.30 mmol) and picolyl chloride (169 mg, 1.03 mmol) in DMF (30 mL). After column chromatography (column Ø3 cm×16 cm) the product was isolated as a white solid. Yield: 394 mg (91%). M.p. 171° C. 1H NMR (400 MHz, CDCl3): δ 0.81 (t, 3JH,H=7.4 Hz, 3H, 4-CH3), 1.31 (s, 9H, 6-CH3), 1.50 (m, 2JH,H=14.5 Hz, 1H, 3-CHA), 1.77 (m, 2JH,H=14.5 Hz, 3JH,H=7.3 Hz, 1H, 3-CHB), 3.86 (s, 3H, 11-CH3), 4.32 (dd, 3JH,H=8.2 Hz, 3JH,H=6.9 Hz, 1H, 2-CH), 4.51 (d, 3JH,H=16.6 Hz, 1H, 12-CHA), 4.71 (d, 3JH,H=16.6 Hz, 1H, 12-CHB), 6.94 (dm, 3JH,H=9.0 Hz, 2H, 8-CH), 7.27 (dd, 3JH,H=7.6 Hz, 3JH,H=4.7 Hz, 1H, 16-CH), 7.72 (dm, 3JH,H=9.0 Hz, 2H, 9-CH), 7.92 (dd, 3JH,H=8.2 Hz, 4JH,H=1.4 Hz, 1H, 17-CH), 8.51 (d, 3JH,H=4.7 Hz, 1H, 15-CH), 8.55 (d, 4JH,H=1.5 Hz, 1H, 14-CH). 13C NMR (100 MHz, CDCl3). δ 11.0 (q, C-4), 24.5 (t, C-3), 27.8 (q, C-6), 46.4 (t, C-12), 55.6 (q, C-11), 62.1 (d, C-2), 82.9 (s, C-5), 114.1 (d, C-8), 123.4 (d, C-16), 129.5 (d, C-9), 131.6 (s, C-7), 134.1 (s, C-13), 136.6 (d, C-17), 148.5 (d, C-15), 149.0 (d, C-14), 162.9 (s, C-10), 167.0 (s, C-1). Exact mass (ESI+): C21H28N2O5S+H+, calcd. 421.1797. found 421.1788; C21H28N2O5S+Na+, calcd. 443.1617. found 443.1609; (C21H28N2O5S)2+H+, calcd. 841.3516. found 841.3514; (C21H28N2O5S)2+Na+, calcd. 863.3336. found 863.3341.
Optical rotation: [α]58920=+44.8, [α]57820=+47.0, [α]54620=+54.0, [α]43620=+98.7, [α]36520=+173.6 (c=1.011, CHCl3).
According to the general procedure methyl (S)—N-(4-methoxyphenylsulfonyl)-aminobutanoate (386 mg, 1.34 mmol) was treated with potassium carbonate (1850 mg, 12.40 mmol) and picolyl chloride (222 mg, 1.34 mmol) in DMF (15 mL). After column chromatography (column Ø2 cm×12 cm) the product was isolated as colorless highly viscose liquid. Yield: 384 mg (76%). 1H NMR (300 MHz, CDCl3). δ 0.80 (t, 3JH,H=7.4 Hz, 3H, 4-CH3), 1.54 (ddq, 2JH,H=14.2 Hz, 3JH,H=7.3 Hz, 3JH,H=7.0 Hz, 1H, 3-CHA), 1.81 (ddq, 2JH,H=14.4 Hz, 3JH,H=7.3 Hz, 3JH,H=7.0 Hz, 1H, 3-CHB), 3.43 (s, 3H, 5-CH3), 3.86 (s, 3H, 10-CH3), 4.43 (dd, 3JH,H=8.1 Hz, 3JH,H=7.0 Hz, 1H, 2-CH), 4.48 (d, 2JH,H=16.5 Hz, 1H, 11-CHA), 4.61 (d, 2JH,H=16.4 Hz, 1H, 11-CHB), 6.96 (d, 3JH,H=9.0 Hz, 2H, 7-CH), 7.25 (dd, 3JH,H=7.8 Hz, 3JH,H=4.9 Hz, 1H, 15-CH), 7.72 (d, 3JH,H=9.0 Hz, 2H, 8-CH), 7.84 (d, 3JH,H=7.9 Hz, 1H, 16-CH), 8.50 (d, 3JH,H=4.5 Hz, 1H, 14-CH), 8.54 (s, 1H, 13-CH). 13C NMR (75.48 MHz, CDCl3): δ 10.8 (q, C-4), 23.9 (t, C-3), 46.4 (t, C-11), 51.9 (q, C-5), 55.6 (q, C-10), 61.2 (d, C-2), 114.0 (d, C-7), 123.3 (d, C-15), 129.5 (d, C-8), 131.2 (s, C-6), 133.5 (s, C-12), 136.4 (d, C-16), 148.7 (d, C-14), 149.1 (d, C-13), 163.0 (s, C-9), 171.2 (s, C-1). Exact mass (ESI+): C18H22N2O5S+H+, calcd. 379.1328. found 379.1325; C18H22N2O5S+Na+, calcd. 401.1147. found 401.1142; (C18H22N2O5S)2+H2, calcd. 757.2577. found 757.2583; (C18H22N2O5S)2+Na+, calcd. 779.2397. found 779.2397. MS (GC-MS-EI): m/z (%) 378 (0) [M+], 349 (3) [M+-C2H5], 319 (100) [M+-CO2CH3], 207 (22) [M+-C7H7O3S], 171 (24) [C7H7O3S+], 147 (13) [C5H9NO2S+], 123 (16) [C7H7O2+], 107 (32) [C7H7O+], 92 (65) [C6H6N+], 77 (14) [C6H5+], 64 (5) [SO2+], 59 (3) [CO2CH3+]. Optical rotation: [α]58920=−52.5, [α]57820=−55.0, [α]54620=−63.4, [α]43620=−115.8, [α]36520=−203.8 (c=1.010, CHCl3).
According to the general procedure methyl (R)—N-(4-methoxyphenylsulfonyl)-aminobutanoate (920 mg, 3.2 mmol), potassium carbonate (4770 mg, 32.0 mmol) and picolyl chloride (539 mg, 3.2 mmol) in DMF (15 mL). After column chromatography (column Ø 3 cm×19 cm) the product was isolated as a colorless highly viscose liquid. Yield: 1160 mg (96%). 1H NMR (400 MHz, CDCl3): δ 0.80 (t, 3JH,H=7.4 Hz, 3H, 4-CH3), 1.51 (m, 2JH,H=21.5 Hz, 3JH,H=7.3 Hz, 1H, 3-CHA), 1.81 (dddd, 2JH,H=21.5 Hz, 3JH,H=14.4 Hz, 3JH,H=7.3 Hz, 3JH,H=7.0 Hz, 1H, 3-CHB), 3.44 (s, 3H, 5-CH3), 3.86 (s, 3H, 10-CH3), 4.44 (dd, 3JH,H=8.4 Hz, 3JH,H=7.0 Hz, 1H, 2-CH), 4.48 (d, 2JH,H=16.6 Hz, 1H, 11-CHA), 4.61 (d, 2JH,H=16.5 Hz, 1H, 11-CHB), 6.96 (dm, 3JH,H=9.0 Hz, 2H, 7-CH), 7.25 (ddd, 3JH,H=7.9 Hz, 3JH,H=4.8 Hz, 4JH,H=0.5 Hz, 1H, 15-CH), 7.72 (dm, 3JH,H=9.0 Hz, 2H, 8-CH), 7.85 (dt, 3JH,H=7.9 Hz, 4JH,H=1.9 Hz, 1H, 16-CH), 8.50 (dd, 3JH,H=4.8 Hz, 4JH,H=1.5 Hz, 1H, 14-CH), 8.53 (d, 4JH,H=1.9 Hz, 1H, 13-CH). 13C NMR (100.62 MHz, CDCl3): δ 10.6 (q, C-4), 23.7 (t, C-3), 46.3 (t, C-11), 51.7 (q, C-5), 55.4 (q, C-10), 61.1 (d, C-2), 113.8 (d, C-7), 123.1 (d, C-15), 129.3 (d, C-8), 131.0 (s, C-6), 133.2 (s, C-12), 136.1 (d, C-16), 148.6 (d, C-14), 149.1 (d, C-13), 162.8 (s, C-9), 170.9 (s, C-1). Exact mass (ESI+): C18H22N2O5S+H+, calcd. 379.1322. found 379.1314; C18H22N2O5S+Na+, calcd. 401.1142. found 401.1135; (C18H22N2O5S)2+H+, cacld. 757.2572. found 757.2559; (C18H22N2O5S)2+Na+, calcd 779.2391. found 779.2383. MS (ESI+, daughter ion experiment): m/z (%) 379 (85) [M++H+], 319 (100) [M+-CO2CH3], 264 (8), 200 (30) [C8H10NO3S+], 171 (22) [C7H7O3S+], 123 (3) [C7H7O2+].
According to the general procedure tert-butyl (S)—N-(4-methoxyphenylsulfonyl)-aminopent-4-enoate (1084 mg, 3.17 mmol), potassium carbonate (4380 mg 31.70 mmol) and picolyl chloride (520 mg, 3.17 mmol) in DMF (25 mL). After column chromatography (column Ø 4 cm×16 cm) the product was isolated as a white solid. Yield: 750 mg (55%). M.p. 176-177° C. 1H NMR (300 MHz, CDCl3): δ 1.31 (s, 9H, 7-CH3), 2.27 (ddd, 2JH,H=14.7 Hz, 3JH,H=8.0 Hz, 3JH,H=6.9 Hz, 1H, 3-CHA), 2.49 (dt, 3JH,H=13.8 Hz, 3JH,H=6.9 Hz, 1H, 3-CHB), 3.86 (s, 3H, 12-CH3), 4.49 (d, 2JH,H=16.1 Hz, 1H, 13-CHA), 4.51 (t, 3JH,H=7.5 Hz, 1H, 2-CH), 4.69 (d, 2JH,H=16.6 Hz, 1H, 13-CHB), 4.89 (dd, 3JH,H=17.1 Hz, 2JH,H=1.5 Hz, 1H, 5-CHtrans), 4.99 (dd, 3JH,H=10.2 Hz, 2JH,H=1.5 Hz, 1H, 5-CHcis), 5.60 (ddt, 3JH,H=17.0 Hz, 3JH,H=10.3 Hz, 3JH,H=6.7 Hz, 1H, 4-CH), 6.94 (dm, 3JH,H=9.0 Hz, 2H, 9-CH), 7.25 (dd, 3JH,H=7.9 Hz, 3JH,H=4.9 Hz, 1H, 17-CH), 7.73 (dm, 3JH,H=9.0 Hz, 2H, 10-CH), 7.86 (d, 3JH,H=7.9 Hz, 1H, 18-CH), 8.50 (d, 3JH,H=4.0 Hz, 1H, 16-CH), 8.54 (d, 4JH,H=1.7 Hz, 1H, 15-CH). 13C NMR (75 MHz, CDCl3): δ 27.8 (q, C-7), 35.5 (t, C-3), 46.6 (t, C-13), 55.6 (q, C-12), 60.2 (d, C-2), 82.3 (s, C-6), 114.1 (d, C-9), 118.4 (t, C-5), 123.3 (d, C-17), 129.6 (d, C-10), 131.5 (s, C-8), 133.0 (d, C-4), 133.7 (s, C-14), 136.6 (d, C-18), 148.5 (d, C-16), 149.0 (d, C-15), 163.0 (s, C-11), 169.3 (s, C-1). Exact mass (ESI+): C22H28N2O5S+H+, calcd. 433.1797. found 433.1812; C22H28N2O5S+Na+, calcd. 455.1617. found 455.1630, (C22H28N2O5S)2+H+, calcd. 865.3516. found 865.3532; (C22H28N2O5S)2+Na+, calcd. 887.3336. found 887.3345. Optical rotation: [α]58920=−26.4, [α]57820=−27.3, [α]54620=−31.2, [α]43620=−56.6, [α]36520=−97.7 (c=1.001, CHCl3):
According to the general procedure tert-butyl (R)—N-(4-methoxyphenylsulfonyl)-aminopent-4-enoate (2375 mg, 6.96 mmol), was treated with potassium carbonate (9600 mg 69.60 mmol) and picolyl chloride (1140 mg 6.96 mmol) in DMF (45 mL). After column chromatography (column æ 5 cm×20 cm) the product was isolated as a white solid. Yield: 2010 mg (67%). M.p. 177° C. 1H NMR (300 MHz, CDCl3): δ 1.31 (s, 9H, 7-CH3), 2.27 (m, 1H, 3-CHA), 2.49 (m, 1H, 3-CHB), 3.86 (s, 3H, 12-CH3), 4.49 (d, 2JH,H=16.4 Hz, 1H, 13-CHA), 4.52 (t, 3, JH,H=7.5 Hz, 1H, 2-CH), 4.69 (d, 2JH,H=16.6 Hz, 1H, 13-CHB), 4.89 (dq, 3JH,H=17.1 Hz, 2JH,H=1.4 Hz, 1H, 5-CHcis), 4.99 (dd, 3JH,H=10.3 Hz, 2JH,H=1.4 Hz, 1H, 5-CHtrans), 5.60 (ddt, 3JH,H=17.0 Hz, 3JH,H=10.3 Hz, 3JH,H=6.7 Hz, 1H, 4-CH), 6.94 (dm, 3JH,H=9.0 Hz, 2 H, 9-CH), 7.26 (ddd, 3JH,H=4.8 Hz, 3JH,H=3.1 Hz, 4JH,H=0.6 Hz, 1H, 17-CH), 7.73 (dm, 3JH,H=9.0 Hz, 2H, 10-CH), 7.87 (dt, 3JH,H=7.9 Hz, 4JH,H=1.8 Hz, 1H, 18-CH), 8.50 (dd, 3JH,H=4.8 Hz, 4JH,H=1.5 Hz, 1H, 16-CH), 8.54 (d, 4JH,H=1.9 Hz, 1H, 15-CH). 13C NMR (75 MHz, CDCl3): δ 27.8 (q, C-7), 35.5 (t, C-3), 46.6 (t, C-13), 55.6 (q, C-12), 60.2 (d, C-2), 82.3 (s, C-6), 114.1 (d, C-9), 118.4 (t, C-5), 123.3 (d, C-17), 129.6 (d, C-10), 131.5 (s, C-8), 133.0 (d, C-4), 133.7 (s, C-14), 136.7 (d, C-18), 148.4 (d, C-16), 149.0 (d, C-15), 163.0 (s, C-11), 169.3 (s, C-1). Exact mass (ESI+): C22H28N2O5S+H+, calcd. 433.1797. found 433.1782; C22H28N2O5S+Na+, calcd. 455.1617. found 455.1599; (C22H28N2O5S)2+H+, calcd. 865.3516. found 865.3505; (C22H28N2O5S)2+Na+, calcd. 887.3336. found 887.3318. MS (ESI+, daughter ion experiment): m/z (%) 433 (0) [M++H+], 377 (12) [433-C4H8], 279 (4) [C13H15N2O3S+], 171 (15) [C7H7O3S+], 165 (100) [C8H9N2O2+], 121 (24) [C7H9N2+], 92 (18) [C6H6N+]. Optical rotation: [α]58920=+26.7, [α]57820=+27.8, [α]54620=+31.8, [α]43620=+57.2, [α]36520=+99.1 (c=1.009, CHCl3).
According to the general procedure tert-butyl (S)—N-(4-methoxyphenylsulfonyl)-2-amino-4-fluorobutanoate (1415 mg, 4.07 mmol), was reacted with potassium carbonate (5500 mg, 40.00 mmol) and picolyl chloride (673 mg 4.1 mmol) in DMF (20 mL). After column chromatographie (column Ø 4 cm×16 cm) the product was isolated as a greenish-white, viscose liquid. Yield: 1512 mg (85%). 1H NMR (300 MHz, CDCl3): δ 1.35 (s, 9H, 6-CH3), 1.86 (m, 1H, 3-CHA), 2.22 (ddt, 2JH,H=13.6 Hz, 3JH,H=9.7 Hz, 3JH,H=6.0 Hz, 1H, 3-CHB), 3.86 (s, 3H, 11-CH3), 4.19-4.45 (m, 2H, 4-CH2F), 4.36 (d, 2JH,H=16.1 Hz, 1H, 12-CHA), 4.49 (dd, 3JH,H=7.7 Hz, 3JH,H=6.6 Hz, 1H, 2-CH), 4.69 (d, 2JH,H=16.2 Hz, 1H, 12-CHB), 6.96 (dm, 3JH,H=8.9 Hz, 2H, 8-CH), 7.25 (dd, 3JH,H=7.9 Hz, 3JH,H=4.8 Hz, 1H, 16-CH), 7.77 (dm, 3JH,H=8.9 Hz, 2H, 9-CH), 7.82 (dm, 3JH,H=8.4 Hz, 1H, 17-CH), 8.51 (s, 1H, 15-CH), 8.53 (s, 1H, 14-CH). 13C NMR (75 MHz, CDCl3): δ 27.7 (q, C-6), 31.8 (dt, 2JH,F=20.5 Hz, C-3), 47.3 (t, C-12), 55.6 (q, C-11), 57.0 (dd, 3Jc=4.3 Hz, C-2), 80.1 (dt, 1JH,F=166.3 Hz, C-4), 82.5 (s, C-5), 114.1 (d, C-8), 123.4 (d, C-16), 129.6 (d, C-9), 131.3 (s, C-7), 132.9 (s, C-13), 136.3 (d, C-17), 149.1 (d, C-15), 149.4 (d, C-14), 163.0 (s, C-10), 169.0 (s, C-1). 19F NMR (282 MHz, CDCl3): δ −221.4 (ddt, 2JH,F=46.9 Hz, 3JH,F=28.3 Hz, 3JH,F=22.8 Hz, 4-CH2F). Exact mass (ESI+): C21H27N2O5S+H+, calcd. 439.1703. found 439.1701; C21H27N2O5S+Na+, calcd. 461.1522. found 431.1526; (C21H27N2O5S)2+Na+, calcd. 877.3328. found 877.3320; (C21H27N2O5S)2+Na+, calcd. 899.3147. found 899.3146. MS (ESI+, daughter ion experiment): m/z (%) 439 (15) [M++H+], 383 (100) [439-C4H8], 171 (5) [C7H7O3S+], 167 (10) [383-C7H7O3S—CO2H], 92 (3) [C6H6N+]. Optical rotation: [α]58920=+34.1, [α]57820=+35.5, [α]54620=+41.1, [α]43620=+74.5, [α]36520=n.d. (c=1.001, CHCl3).
According to the general procedure tert-butyl (S)—N-(4-methoxyphenyl-sulfonyl)-2-amino-4-fluorobutanoate (500 mg, 1.44 mmol) was reacted with potassium carbonate (1990 mg 14.40 mmol) and picolyl chloride (237 mg, 1.44 mmol) in DMF (20 mL). After column chromatography (column Ø 2 cm×15 cm) the product was isolated as greenish-white, viscose liquid. Yield: 587 mg (93%). 1H NMR (400 MHz, CDCl3): δ 1.35 (s, 9H, 6-CH3), 1.86 (m, 3JH,F=28.3 Hz, 2JH,H=14.2 Hz, 1H, 3-CHB), 2.22 (dddt, 3JH,F=22.7 Hz, 2JH,H=14.2 Hz, 3JH,H=7.5 Hz, 3JH,H=6.6 Hz, 1H, 3-CHA), 3.87 (s, 3H, 11-CH3), 4.37 (d, 2JH,H=16.2 Hz, 1H, 12-CHA), 4.22-4.46 (m, 2JH,F=47.1 Hz, 2H, 4-CH2F), 4.48 (dd, 3JH,H=7.9 Hz, 3JH,H=6.7 Hz, 1H, 2-CH), 4.69 (d, 2JH,H=16.2 Hz, 1H, 12-CHB), 6.96 (dm, 3JH,H=9.0 Hz, 2H, 8-CH), 7.25 (dd, 3JH,H=7.9 Hz, 3JH,H=4.8 Hz, 1H, 16-CH), 7.77 (dm, 3JH,H=8.9 Hz, 2H, 9-CH), 7.82 (dt, 3JH,H=7.9 Hz, 4JH,H=1.8 Hz, 1H, 17-CH), 8.50 (d, 4JH,H=2.0 Hz, 1H, 14-CH), 8.52 (dd, 3JH,H=4.8 Hz, 4JH,H=1.6 Hz, 1H, 15-CH). 13C NMR (100 MHz, CDCl3): δ 27.8 (q, C-6), 31.9 (dt, 2JH,F=20.5 Hz, C-3), 47.3 (t, C-12), 55.6 (q, C-11), 57.1 (dd, 3JH,F=4.3 Hz, C-2), 80.2 (dt, 1JH,F=166.4 Hz, C-4), 82.6 (s, C-5), 114.2 (d, C-8), 123.4 (d, C-16), 129.7 (d, C-9), 131.4 (s, C-7), 132.9 (s, C-13), 136.3 (d, C-17), 149.2 (d, C-15), 149.5 (d, C-14), 163.1 (s, C-10), 169.1 (s, C-1). 19F NMR (282 MHz, CDCl3): δ 221.4 (tdd, 2JH,F=46.9 Hz, 3JH,F=28.3 Hz, 3JH,F=22.8 Hz, 4-CH2F). Elemental analysis: C21H27FN2O5S (M=438.513 g/mol); calcd. C, 57.52; H, 6.21, N, 6.39. found C, 57.82; H, 6.00; N, 6.38%. Exact mass (ESI+): C21H27FN2O5S+H+, calcd. 439.1703. found 439.1697; C21H27FN2O5S+Na+, calcd. 461.1522. found 461.1513; (C21H27FN2O5S)2+H+, calcd. 877.3328. found 877.3321; (C21H27FN2O5S)2+Na+, calcd. 899.3147. found 899.3145. MS (GC/MS, 70 eV): m/z (%) 438 (0) [M+], 362 (1), 277 (3) [M+-C8H14FO2], 191 (100), 171 (22) [C7H7O3S+], 147 (75), 123 (12) [C6H4OS+], 107 (22) [C7H7O+], 92 (32) [C6H6N+], 77 (10) [C6H5+]. Optical rotation: [α]58920=−33.2, [α]57820=−34.5, [α]54620=−39.9, [α]43620=−72.6, [α]36520=−124.9 (c=1.001, CHCl3).
According to the general procedure tert-butyl (S)—N-(4-methoxyphenyl-sulfonyl)-2-amino-4-fluoropent-4-enoate (1500 mg, 4.17 mmol) was reacred with potassium carbonate (5760 mg 41.7 mmol) and picolyl chloride (685 mg, 4.17 mmol) in DMF (20 mL). After column chromatography (column Ø 4 cm×16 cm) the product was isolated as yellow, viscose liquid. Yield: 1428 mg (76%). 1H NMR (300 MHz, CDCl3): δ 1.35 (s, 9H, 7-CH3), 2.48 (m, 3JH,F=19.2 Hz, 1H, 3-CHA), 2.73 (td, 3JH,F=15.0 Hz, 3JH,H=6.8 Hz, 1H, 3-CHB), 3.86 (s, 3H, 12-CH3), 4.14 (dd, 3JH,F=49.7 Hz, 2JH,H=3.1 Hz, 1H, 5-CHtrans), 4.39 (d, 2JH,H=16.3 Hz, 1H, 13-CHA), 4.55 (dd, 3JH,F=17.2 Hz, 2JH,H=3.0 Hz, 1H, 5-CHcis), 4.58 (t, 3JH,H=7.3 Hz, 1H, 2-CH), 4.65 (d, 2JH,H=16.3 Hz, 1H, 13-CHB), 6.95 (d, 3JH,H=9.0 Hz, 2H, 9-CH), 7.23 (ddd, 3JH,H=7.9 Hz, 3JH,H=4.8 Hz, 4JH,H=0.7 Hz, 1H, 17-CH), 7.78 (m, 1H, 18-CH), 7.78 (d, 3JH,H=9.0 Hz, 2H, 10-CH), 8.51 (dd, 3JH,H=4.6 Hz, 4JH,H=1.7 Hz, 2H, 15-CH and 16-CH). 13C NMR (75 MHz, CDCl3): δ 27.8 (s, C-7), 34.0 (d, 2JH,F=28.1 Hz, C-3), 47.4 (s, C-13), 55.6 (s, C-12), 57.6 (s, C-2), 82.8 (s, C-6), 93.4 (d, 2JH,F=19.1 Hz, C-5), 114.2 (s, C-9), 123.3 (s, C-17), 129.8 (s, C-10), 131.4 (s, C-8), 132.8 (s, C-14), 136.4 (s, C-18), 149.1 (s, C-16), 149.5 (s, C-15), 161.5 (d, 1JH,F=256.5 Hz, C-4), 163.1 (s, C-11), 168.4 (s, C-1). 19F NMR (282 MHz, CDCl3): δ −97.6 (dddd, 3JH,F=50.0 Hz, 3JH,F=19.2 Hz, 3JH,F=17.2 Hz, 3JH,F=15.0 Hz, 4-CF). Exact mass (ESI4): C22H27FN2O5S+Na+, cacld. 451.1703. found 451.1701; C22H27FN2O5S+Na+, calcd. 473.1522. found 473.1521; C22H27FN2O5S)2+H+, calcd. 901.3328. found 901.3320; (C22H27FN2O5S)2+Na+, calcd. 923.3147. found 923.3144.
Optical rotation: [α]58920=−6.3, [α]57820=−6.8, [α]54620=−7.9, [α]43620=−14.2, [α]36520=−22.9 (c=1.010, CHCl3).
According to the general procedure tert-butyl (R)—N-(4-methoxyphenyl-sulfonyl)-2-amino-4-fluoropent-4-enoate (720 mg, 2.00 mmol) was reacted with potassium carbonate (2760 mg, 20.00 mmol) and picolyl chloride (329 mg, 2.00 mmol) in DMF (20 mL). After column chromatography the product was isolated as yellow, viscose oil. Yield: 523 g (58%). 1H NMR (500 MHz, CDCl3): δ 1.37 (s, 9H, 7-CH3), 2.48 (ddd, 3JH,F=19.2 Hz, 2JH,H=15.1 Hz, 3JH,H=7.8 Hz, 1H, 3-CHA), 2.74 (dt, 2JH,H=15.2 Hz, 3JH,H=6.9 Hz, 1H, 3-CHB), 3.88 (s, 3 H, 12-CH3), 4.17 (dd, 3JH,F=49.7 Hz, 2JH,H=3.1 Hz, 1H, 5-CHtrans), 4.42 (d, 2JH,H=16.3 Hz, 1H, 13-CHA), 4.57 (dd, 3JH,F=17.2 Hz, 2JH,H=3.1 Hz, 1H, 5-CHcis), 4.60 (t, 3JH,H=7.3 Hz, 1H, 2-CH), 4.67 (d, 2JH,H=16.3 Hz, 1H, 13-CHB), 6.97 (dm, 3JH,H=9.0 Hz, 2H, 9-CH), 7.29 (dd, 3JH,H=7.9 Hz, 3JH,H=4.9 Hz, 1H, 17-CH), 7.79 (dm, 3JH,H=9.0 Hz, 2H, 10-CH), 7.86 (dd, 3JH,H=7.8 Hz, 4JH,H=1.4 Hz, 1H, 18-CH), 8.52-8.54 (m, 4JH,H=1.5 Hz, 2H, 15-CH and 16-CH). 13C NMR (100.62 MHz, CDCl3): δ 27.8 (q, C-7), 34.0 (dt, 2JH,F=28.1 Hz, C-3), 47.3 (t, C-13), 55.6 q, C-12), 57.6 (d, C-2), 82.9 (s, C-6), 93.4 (dt, 2JH,F=19.0 Hz, C-5), 114.2 (d, C-9), 123.4 (d, C-17), 129.8 (d, C-10), 131.4 (s, C-8), 132.9 (s, C-14), 136.6 (d, C-18), 148.9 (d, C-16), 149.3 (d, C-15), 161.5 (ds, 1JH,F=256.5 Hz, C-4), 163.1 (s, C-11), 168.5 (s, C-1). 19F NMR (470 MHz, CDCl3): δ −97.6 (dddd, 3JH,F=49.7 Hz, 3JH,F=19.2 Hz, 3JH,F=17.2 Hz, 3JH,F=15.0 Hz, 4-CF).
Optical rotation: [α]58920=+7.7, [α]57820=+8.1, [α]54620=+9.3, [α]43620=+16.3, [α]36520=+28.4 (c=1.004, CHCl3).
General Procedure for Hydrolysis of Amino Acid Tert-Butylesters
In a dried YOUNG-tube trifluoroacetic acid (20 mL/mmol) is added under argon to the corresponding amino acid tert-butylester dissolved in dry dichloromethane (20 mL/mmol). After flushing with argon the YOUNG-tube is sealed and the mixture is stirred at r.t. for 3-4 h. Then the reaction mixture was evaporated to dryness. The residue is dissolved in chloroform (100 mL/mmol) and washed with an aqueous solution of citric acid and bicarbonate (pH≈4) (25 mL/mmol). The aqueous phase is extracted with chloroform (4×30 mL/mmol) and the combined organic layer is dried with magnesium sulfate. The solvent is evaporated to get the crude product.
wherein R3 is C1-C6-fluoro-alkyl or optionally substituted C2-C6 alkenyl wherein the substituent is a F.
According to the general procedure tert-butyl (S)—N-(4-methoxyphenylsulfonyl)-N-(3-pyridylmethyl)-aminobutanoate (243 mg, 0.58 mmol) was dissolved in dry dichloromethane (12 mL) and treated with trifluoroacetic acid (12 mL). The product was isolated as colorless, highly viscos oil. Yield: 194 mg (91%). 1H NMR (300 MHz, DMSO-d6): δ 0.70 (t, 3JH,H=7.3 Hz, 3H, 4-CH3), 1.45 (ddd, 2JH,H=14.2 Hz, 3JH,H=8.7 Hz, 3JH,H=7.3 Hz, 1H, 3-CHA), 1.75 (dt, 2JH,H=14.0 Hz, 3JH,H=6.8 Hz, 1H, 3-CHB), 3.83 (s, 3H, 9-CH3), 4.23 (dd, 3JH,H=8.9 Hz, 3JH,H=6.0 Hz, 1H, 2-CH), 4.43 (d, 2JH,H=16.9 Hz, 1H, 10-CHA), 4.61 (d, 2JH,H=16.9 Hz, 1H, 10-CHB), 7.07 (dm, 3JH,H=9.0 Hz, 2H, 6-CH), 7.38 (dd, 3JH,H=7.8 Hz, 3JH,H=4.9 Hz, 1H, 14-CH), 7.74 (dm, 3JH,H=9.0 Hz, 2H, 7-CH), 7.84 (d, 3JH,H=7.9 Hz, 1H, 15-CH), 8.47 (d, 3JH,H=4.1 Hz, 1H, 13-CH), 8.57 (s, 1H, 12-CH). 13C NMR (75 MHz, DMSO-d6): δ 11.0 (q, C-4), 23.2 (t, C-3), 46.4 (t, C-10), 55.7 (q, C-9), 61.5 (d, C-2), 114.3 (d, C-6), 123.4 (d, C-14), 129.5 (d, C-7), 131.0 (s, C-5), 134.3 (s, C-11), 136.3 (d, C-15), 148.0 (d, C-13), 148.9 (d, C-12), 162.6 (s, C-8), 171.7 (s, C-1). Exact mass (ESI+): C17H20N2O5S+H+, calcd. 365.1171. found 365.1169; C17H20N2O5S+Na+, calcd. 387.0991. found 387.0988; (C17H20N2O5S)2+H+, calcd. 729.2264. found 729.2267; (C17H20N2O5S)2+Na+, calcd. 751.2084. found 751.2085. MS (ESI+, daughter ion experiment): m/z (%) 365 (90) [M++H+], 319 (100) [365−H+—CO21-1], 280 (65) [C13H16N2O3S+], 228 (15) [319-C6H6N], 200 (30) [228-C2H5], 171 (55) [C7H7O3S+].
According to the general procedure tert-butyl (R)—N-(4-methoxyphenylsulfonyl)-N-(3-pyridylmethyl)-aminobutanoate (133 mg, 0.32 mmol) dissolved in dry dichloromethane (8 mL) and treated with trifluoroacetic (8 mL) acid. The product was isolated as colorless, highly viscos oil. Yield: 112 mg (96%). 1H NMR (300 MHz, DMSO-d6): δ 0.71 (t, 3JH,H=7.3 Hz, 3 H, 4-CH3), 1.59-1.87 (m, 2H, 3-CH2), 3.84 (s, 3H, 9-CH3), 4.25 (dd, 3JH,H=8.8 Hz, 3JH,H=6.0 Hz, 1H, 2-CH), 4.52 (d, 2JH,H=17.1 Hz, 1H, 10-CHA), 4.66 (d, 2JH,H=17.1 Hz, 1H, 10-CHB), 7.08 (dm, 3JH,H=8.9 Hz, 2H, 6-CH), 7.65 (m, 1H, 14-CH), 7.76 (dm, 3JH,H=8.9 Hz, 2H, 7-CH), 8.10 (d, 3JH,H=7.8 Hz, 1H, 15-CH), 8.45-8.95 (m, 2H, 12-CH & 13-CH), 12.44 (s br, 1H, 16-OH). 13C-NMR (75 MHz, DMSO-d6): δ 11.0 (q, C-4), 23.2 (t, C-3), 46.2 (t, C-10), 55.7 (q, C-9), 61.5 (d, C-2), 114.3 (d, C-6), 124.6 (d, C-14), 129.5 (d, C-7), 130.4 (s, C-11), 130.8 (s, C-5), 139.0 (d, C-15), 145.5 (d, C-13), 146.2 (d, C-12), 162.7 (s, C-8), 171.7 (s, C-1). Exact mass (ESI+): C17H20N2O5S+H+, calcd. 365.1171, 365.1166; C17H20N2O5S+Na+, calcd. 387.0991. found 387.0986; (C17H20N2O5S)2+H+, calcd. 729.2264. found 729.2267; (C17H20N2O5S)2+Na+, calcd. 751.2084. found 751.2085. MS (ESI+, daughter ion experiment): m/z (%) 365 (100) [M++H+], 319 (32) [365-H+—CO2H], 280 (20) [C13H16N2O3S+], 228 (5) [319-C6H6N], 200 (10) [228-C2H5], 171 (18) [C7H7O3S+], 149 (38) [319-C7H7O3S].
According to the general procedure tert-butyl (S)—N-(4-methoxyphenylsulfonyl)-N-(3-pyridylmethyl)-2-aminopent-4-enoate (400 mg, 0.93 mmol) was dissolved in dry dichloromethane (20 mL) and treated with trifluoroacetic acid (20 mL). The product was isolated as colorless, highly viscos oil. Yield: 312 mg (89%). 1H NMR (400 MHz, DMSO-d6): δ 2.16 (ddd, 2JH,H=15.0 Hz, 3JH,H=8.4 Hz, 3JH,H=7.4 Hz, 1H, 3-CHA), 2.42 (m, 1H, 3-CHB), 3.76 (s, 3H, 10-CH3), 4.35 (dd, 3JH,H=8.7 Hz, 2JH,H=6.2 Hz, 1H, 2-CH), 4.36 (d, 2JH,H=17.3 Hz, 1H, 11-CHA), 4.51 (d, 2JH,H=16.9 Hz, 1H, 11-CHB), 4.80 (dd, 3JH,H=17.2 Hz, 2JH,H=1.6 Hz, 1H, 5-CHtrans), 4.84 (dd, 3JH,H=10.3 Hz, 2JH,H=1.5 Hz, 1H, 5-CHcis), trans, 5.49 (ddt, 3JH,H=17.0 Hz, 3JH,H=10.3 Hz, 3JH,H=6.7 Hz, 1H, 4-CH), 6.99 (dm, 3JH,H=9.0 Hz, 2H, 7-CH), 7.25 (dd, 3JH,H=7.8 Hz, 3JH,H=4.8 Hz, 1H, 15-CH), 7.66 (dm, 3JH,H=8.9 Hz, 2H, 8-CH), 7.70 (m, 1H, 16-CH), 8.36 (d, 3JH,H=4.1 Hz, 1H, 14-CH), 8.46 (s, 1H, 13-CH). 13C NMR (100 MHz, DMSO-d6): δ 34.1 (t, C-3), 46.5 (t, C-11), 55.7 (q, C-10), 59.7 (d, C-2), 114.3 (d, C-7), 117.8 (t, C-5), 123.1 (d, C-15), 129.5 (d, C-8), 131.0 (s, C-6), 133.8 (d, C-4), 134.0 (s, C-12), 135.9 (d, C-16), 148.1 (d, C-14), 149.2 (d, C-13), 162.6 (s, C-9), 171.2 (s, C-1). Exact mass (ESI+): C18H20N2O5S+H+, calcd. 377.1171. found 377.1186; C18H20N2O5S+Na+, calcd. 399.0991. found 399.1004; (C18H20N2O5S)2+H+, calcd. 753.2264. found 753.2259; (C18H20N2O5S)2+Na+, calcd. 775.2084. found 775.2094. Exact mass (ESI−): C18H20N2O5S—H+, calcd. 375.1015. found 375.1035; (C18H20N2O5S)-2—H+, calcd. 751.2108. found 751.2141. MS (ESI+, daughter ion experiment): m/z (%) 377 (75) [M++H+], 331 (75) [377-H+—CO2H], 280 (42) [C13H16N2O3S+], 240 (8) [331-C6H6N], 200 (20) [C8H10NO3S+], 171 (50) [C7H7O3S+], 161 (100) [331-C7H7O3S], 123 (8) [C7H7O2+].
According to the general procedure tert-butyl (R)—N-(4-methoxyphenylsulfonyl)-N-(3-pyridylmethyl)-2-aminopent-4-enoate (247 mg, 0.57 mmol) was dissolved in dry dichloromethane (12 mL) and treated with trifluoroacetic acid (12 mL). The product was isolated as white highly viscos oil. Yield: 196 mg (92%). 1H NMR (400 MHz, DMSO-d6): δ 2.29 (ddd, 2JH,H=15.0 Hz, 3JH,H=8.6 Hz, 3JH,H=7.6 Hz, 1H, 3-CHA), 2.54 (m, 2JH,H=15.0 Hz, 3JH,H=6.1 Hz, 1H, 3-CHB), 3.84 (s, 3H, 10-CH3), 4.48 (dd, 3JH,H=8.9 Hz, 3JH,H=6.0 Hz, 1H, 2-CH), 4.53 (d, 2JH,H=17.3 Hz, 1H, 11-CHA), 4.65 (d, 2JH,H=17.2 Hz, 1H, 11-CHB), 4.89 (dd, 3JH,H=17.2 Hz, 2JH,H=1.7 Hz, 1H, 5-CHtrans), 4.93 (dd, 3JH,H=10.5 Hz, 2JH,H=1.6 Hz, 1H, 5-CHcis), 5.58 (ddt, 3JH,H=17.1 Hz, 3JH,H=10.3 Hz, 3JH,H=6.7 Hz, 1H, 4-CH), 7.09 (d, 3JH,H=9.0 Hz, 2H, 7-CH), 7.58 (dd, 3JH,H=7.9 Hz, 3JH,H=5.1 Hz, 1H, 15-CH), 7.75 (d, 3JH,H=9.0 Hz, 2H, 8-CH), 8.07 (d, 3JH,H=8.0 Hz, 1H, 16-CH), 8.59 (d, 3JH,H=4.4 Hz, 1H, 14-CH), 8.67 (s, 1H, 13-CH), 12.82 (s br, 1H, 17-OH). 13C NMR (100 MHz, DMSO-d6): δ 34.0 (t, C-3), 46.3 (t, C-11), 55.7 (q, C-10), 59.7 (d, C-2), 114.3 (d, C-7), 117.8 (t, C-5), 124.3 (d, C-15), 129.6 (d, C-8), 130.7 (s, C-6), 134.0 (d, C-4), 135.7 (s, C-12), 139.1 (d, C-16), 145.6 (d, C-14), 146.4 (d, C-13), 162.7 (s, C-9), 171.2 (s, C-1).
According to the general procedure tert-butyl (S)—N-(4-methoxyphenylsulfonyl)-N-(3-pyridylmethyl)-2-amino-4-fluorobutanoate (562 mg, 1.28 mmol) was dissolved in dry dichloromethane (5 mL) and treated with trifluoroacetic acid (25 mL). The product was isolated as white solid. Yield: 472 mg (96%). M.p. 156° C. 1H NMR (300 MHz, DMSO-d6): δ 1.85 (m, 3JH,F=22.3 Hz, 1H, 3-CHA), 2.19 (m, 3JH,F=27.9 Hz, 1H, 3-CHB), 3.84 (s, 3H, 9-CH3), 4.32 (dt, 2JH,F=47.1 Hz, 3JH,H=5.7 Hz, 2H, 4-CH2), 4.43 (d, 2JH,H=17.2 Hz, 1H, 10-CHA), 4.49 (m, 1H, 2-CH), 4.62 (d, 2JH,H=16.8 Hz, 1H, 10-CHB), 7.09 (dm, 3JH,H=8.9 Hz, 2H, 6-CH), 7.39 (dd, 3JH,H=7.8 Hz, 3JH,H=4.8 Hz, 1H, 14-CH), 7.75 (dm, 3JH,H=8.9 Hz, 2H, 7-CH), 7.82 (m, 1H, 15-CH), 8.49 (d, 3JH,H=3.7 Hz, 1H, 13-CH), 8.55 (s, 1H, 12-CH), 13.02 (s br, 1H, 16-OH). 13C NMR (75 MHz, DMSO-d6): δ 30.9 (dt, 2JH,F=20.2 Hz, C-3), 46.8 (t, C-10), 55.7 (q, C-9), 56.3 (dd, 3JH,F=5.0 Hz, C-2), 80.4 (dt, 1JH,F=163.1 Hz, C-4), 114.4 (d, C-6), 123.5 (d, C-14), 129.6 (d, C-7), 130.8 (s, C-5), 133.9 (s, C-11), 136.4 (d, C-15), 148.0 (d, C-13), 148.8 (d, C-12), 162.7 (s, C-8), 171.2 (s, C-1). 19F NMR (282 MHz, DMSO-d6): δ −219.3 (ddt, 2JH,F=47.1 Hz, 3JH,F=27.8 Hz, 3JH,F=22.3 Hz, 4-CH2F). Exact mass (ESI+): C17H19FN2O5S+H+, calcd. 383.1077. found 383.1073; C17H19FN2O5S+Na+. found 405.0896. found 405.0892; (C17H19FN2O5S)2+H+, calcd. 765.2076. found 765.2071; (C17H19FN2O5S)2+Na+, calcd. 787.1895. found 787.1887. Exact mass (ESI−): C17H19FN2O5S—H+, calcd. 381.0926. found 381.0930; (C17H19FN2O5S—H)2+H+, calcd. 763.1925. found 763.1930; (C17H19FN2O5S—H+)2+Na+, calcd. 785.1744. found 785.1758.
According to the general procedure tert-butyl (R)—N-(4-methoxyphenylsulfonyl)-N-(3-pyridylmethyl)-2-amino-4-fluorobutanoate (448 mg, 1.02 mmol) was dissolved in dry dichloromethane (25 mL) and treated with trifluoroacetic acid (25 mL). The product was isolated as white solid. Yield: 352 mg (90%). M.p. 155-156° C. 1H NMR (400 MHz, DMSO-d6): δ 1.79 (m, 3JH,F=22.8 Hz, 1H, 3-CHA), 2.19 (m, 3JH,F=27.6 Hz, 1H, 3-CHB), 3.84 (s, 3H, 9-CH3), 4.32 (t, 2JH,H=5.7 Hz, 1H, 4-CHA), 4.37 (t, 2JH,H=5.7 Hz, 1H, 4-CHB), 4.41 (d, 2JH,H=17.1 Hz, 1H, 10-CHA), 4.48 (t, 3JH,H=7.2 Hz, 1H, 2-CH), 4.61 (d, 2JH,H=16.7 Hz, 1H, 10-CHB), 7.09 (dm, 3JH,H=8.5 Hz, 2H, 6-CH), 7.34 (dd, 3JH,H=7.6 Hz, 3JH,H=4.9 Hz, 1H, 14-CH), 7.72-7.80 (m, 3H, 7-CH and 15-CH), 8.46 (d, 3JH,H=4.7 Hz, 1 H, 13-CH), 8.52 (s, 1H, 12-CH), 13.11 (s br, 1H, 16-OH). 13C NMR (100 MHz, DMSO-d6): δ 31.0 (dt, 2JH,F=20.2 Hz, C-3), 46.8 (t, C-10), 55.7 (q, C-9), 56.2 (dd, 3JH,F=5.0 Hz, C-2), 80.4 (dt, 1JH,F=163.1 Hz, C-4), 114.3 (d, C-6), 123.2 (d, C-14), 129.5 (d, C-7), 130.9 (s, C-5), 133.6 (s, C-11), 135.8 (d, C-15), 148.5 (d, C-13), 149.3 (d, C-12), 162.7 (s, C-8), 171.2 (s, C-1). 19F NMR (282 MHz, DMSO-d6): δ −221.8 (ddt, 2JH,F=47.1 Hz, 2JH,F=27.5 Hz, 2JH,F=22.5 Hz, 4-CH2F). Exact mass (ESI+): C17H19FN2O5S+H+, calcd. 383.1077. found 383.1079; C17H19FN2O5S+Na+, calcd. 405.0896. found 405.0900; (C17H19FN2O5S)2+H+, calcd. 765.2076. found 765.2071; (C17H19FN2O5S)2+Na+, calcd. 787.1895. found 787.1884. MS (ESI+, daughter ion experiment): m/z (%) 383 (75) [M++H+], 363 (4) [M+-HF], 337 (22) [383-H+—CO2H], 280 (40) [C13H16N2O3S+], 211 (5) [383-C7H7O3S], 200 (13) [C8H10NO3S+], 171 (18) [C7H7O3S+], 167 (100) [211-CO2H], 123 (8) [C7H7O2+].
According to the general procedure tert-butyl (S)—N-(4-Methoxyphenylsulfonyl)-N-(3-pyridylmethyl)-2-amino-4-fluoropent-4-enoate (272 mg, 0.60 mmol) was dissolved in dry dichloromethane (20 mL) and treated with 20 mL trifluoroacetic acid. The product was isolated as yellowish-white solid. Yield: 200 mg (85%). M.p. 172° C. 1H NMR (300 MHz, DMSO-d6): δ 2.59 (m, 1H, 3-CHA), 2.84 (ddd, 3JH,F=15.7 Hz, 2JH,H=12.6 Hz, 3JH,H=5.5 Hz, 1H, 3-CHB), 3.91 (s, 3H, 10-CH3), 4.30 (dd, 3JH,F=51.6 Hz, 2JH,H=3.1 Hz, 1H, 4.53 (d, 2JH,H=16.9 Hz, 1H, 11-CHA), 4.62 (dd, 3JH,F=17.8 Hz, 2JH,H=3.0 Hz, 1H, 5-CHcis), 4.66 (d, 2JH,H=16.8 Hz, 1H, 11-CHB), 4.69 (dd, 3JH,H=5.7 Hz, 3JH,H=3.0 Hz, 1H, 2-CH), 7.14 (d, 3JH,H=9.1 Hz, 2H, 7-CH), 7.45 (m, 1H, 15-CH), 7.80 (d, 3JH,H=9.0 Hz, 2 H, 8-CH), 7.88 (m, 1H, 16-CH), 8.55 (s, 1H, 14-CH), 8.63 (s, 1H, 13-CH), 12.94 (s, 1H, 17-OH). 13C NMR (75 MHz, DMSO-d6): δ 32.6 (dt, 2JH,F=28.0 Hz, C-3), 46.7 (t, C-11), 55.7 (q, C-10), 57.0 (d, C-2), 93.2 (dt, 2JH,F=18.4 Hz, C-5), 114.3 (d, C-7), 123.4 (d, C-15), 129.4 (d, C-8), 129.6 (s, C-6), 130.7 (s, C-12), 136.6 (d, C-16), 147.7 (d, C-14), 148.7 (d, C-13), 161.7 (ds, 1JH,F=255.3 Hz, C-4), 162.7 (s, C-9), 169.9 (s, C-1). 19F NMR (282 MHz, DMSO-d6): δ −96.2 (m, 3JH,F=51.5 Hz, 3JH,F=17.8 Hz, 3JH,F=15.7 Hz, 4-CF). Exact mass (ESI+): C18H19FN2O5S+H+, calcd. 395.1077. found 395.1088; C18H19FN2O5S+Na+, calcd. 417.0896. found 417.0905; (C18H19FN2O5S)2+H+, calcd. 789.2076. found 789.2096; (C18H19FN2O5S)2+Na+, calcd. 811.1895. found 811.1897. Exact mass (ESI−): C18H19FN2O5S—H+, calcd. 393.0920. found 393.0940; (C18H19FN2O5S—H+)2+H+, calcd. 787.1919. found 787.1955; (C18H19FN2O5S—H+)2+Na+, calcd. 809.1739. found 809.1766. MS (ESI+, daughter ion experiment): m/z (%) 395 (12) [M++H+], 179 (22) [M+-C2H2F—C7H7O3S], 171 (31) [C7H7O3S+], 165 (100) [M+-C3H4F—C7H7O3S], 147 (10), 123 (8), 107 (18) [C7H7O+], 92 (20) [C6H7N+]. MS (ESI−, daughter ion experiment): m/z (%) 393 (2) [M−-H+], 277 (27) [393-C5H5FO2−], 171 (100) [C7H7O3S−], 157 (4) [C6HSO3S−], 115 (18) [C5H4FO2−].
According to the general procedure tert-butyl (R)—N-(4-methoxyphenylsulfonyl)-N-(3-pyridylmethyl)-2-amino-4-fluoropent-4-enoate (256 mg, 0.57 mmol) was dissolved in dry dichloromethane (15 mL) and treated with trifluoroacetic acid (15 mL). The product was isolated as yellowish-white solid. Yield: 204 mg (91%). M.p. 172-173° C. 1H NMR (300 MHz, DMSO-d6): δ 2.51 (m, 1H, 3-CHA), 2.77 (ddd, 3JH,F=21.2 Hz, 2JH,H=12.7 Hz, 3JH,H=5.8 Hz, 1H, 3-CHB), 4.22 (dd, 3JH,F=51.5 Hz, 2JH,H=3.0 Hz, 1H, 5-CHtrans), 4.45 (d, 2JH,H=16.8 Hz, 1H, 11-CHA), 4.55 (dd, 3JH,F=17.8 Hz, 2JH,H=3.0 Hz, 1H, 5-CHcis), 4.59 (d, 2JH,H=16.5 Hz, 1H, 11-CHB), 4.62 (t, 3JH,H=4.3 Hz, 1H, 2-CH), 7.07 (d, 3JH,H=9.0 Hz, 2H, 7-CH), 7.33 (dd, 3JH,H=7.7 Hz, 3JH,H=4.8 Hz, 1H, 15-CH), 7.73 (d, 3JH,H=9.0 Hz, 2H, 8-CH), 7.79 (m, 1H, 16-CH), 8.45 (d, 3JH,H=4.6 Hz, 1H, 14-CH), 8.54 (s, 1H, 13-CH), 12.89 (s br, 1H, 17-OH). 13C NMR (75.48 MHz, DMSO-d6): δ 32.7 (dt, 2JH,F=27.6 Hz, C-3), 46.7 (t, C-11), 55.7 (q, C-10), 56.9 (d, C-2), 93.2 (dt, 2JH,F=18.1 Hz, C-5), 114.3 (d, C-7), 123.2 (d, C-15), 129.6 (d, C-8), 130.8 (s, C-6), 132.8 (s, C-12), 136.1 (d, C-16), 148.1 (d, C-14), 149.1 (d, C-13), 161.7 (ds, 1JH,F=255.3 Hz, C-4), 162.7 (s, C-9), 170.5 (s, C-1). 19F NMR (282.37 MHz, DMSO-d6): δ −96.2 (dddd, 3JH,F=51.5 Hz, 3JH,F=21.1 Hz, 3JH,F=17.9 Hz, 3JH,F=12.6 Hz, 4-CF). Exact mass (ESI+): C18H19FN2O5S+H+, calcd. 395.1077. found 395.1080; C18H19FN2O5S+Na+, calcd. 417.0896. found 417.0896. Exact mass (ESI−): C18H19FN2O5S—H+, calcd. 393.0920. found 393.0948; (C18H19FN2O5S)2—H+, calcd. 787.1919. found 787.1940. MS (ESI+, daughter ion experiment): m/z (%) 395 (45) [M++H+], 375 (5) [395-HF], 349 (22) [395-H+—CO2H], 305 (20) [349-C2H2F], 280 (30) [C13H16N2O3S+], 200 (10) [305-C7H7O], 179 (100) [C6H10FNO2S+], 171 (75) [C7H7O3S+].
General Procedure for Hydrolysis of Amino Acid Methylesters
The aminoacid methylester (1 equivalent) dissolved in a mixture of THF, methanol and water (ratio 3:1:1; 3.3 mL/mmol) is treated with lithium hydroxide (5 equivalents). This solution is stirred at r.t. over night. The reaction mixture is extracted with ethyl acetate (2×20 mL). This extract is discharged. Then the aqueous phase is acidified with 0.5 N hydrochloric acid and extracted again with ethyl acetate (4×20 mL). The combined organic phases are dried with magnesium sulfate and the solvent is evaporated.
According to the general procedure methyl (S)—N-(4-methoxyphenylsulfonyl)-N-(3-pyridylmethyl)-aminobutanoate (306 mg, 0.81 mmol) was dissolved in the above-mentioned solvent mixture (10 mL) and treated with lithium hydroxide (169 mg, 4.04 mmol). The product was isolated as white, highly viscos liquid. Yield: 174 mg (59%). The spectroscopic data agree with those given above.
According to the general procedure methyl (R)—N-(4-methoxyphenylsulfonyl)-N-(3-pyridylmethyl)-aminobutanoate (420 mg, 1.11 mmol) was dissolved in the above-mentioned solvent mixture (10 mL) and treated with lithium hydroxide (230 mg, 5.55 mmol). The product was isolated as white, highly viscos liquid. Yield: 251 mg (62%). The spectroscopic data agree with those given above.
Synthesis of the Hydroxamic Acids
General Procedure for Synthesis of the Hydroxamates
The N,N-disubstituted α-amino acid (1 equivalent) is suspended in dichloromethane (15 mL/mmol). Then 1-hydroxy-benzotriazole (HOBT, 1 equivalent) is added and stirred for a short time. After addition of N-methylmorpholin (NMM) (5 equiv.) the acid is dissolved. Subsequently, O-tert-butylhydroxylamine (3 equiv.) was added and the solution is stirred for 5 min 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide (EDC) hydrochloride (1.3 equiv.) is added and the reaction mixture is stirred overnight. The solution is washed with water (20 mL) and the aqueous phase is extracted with dichloromethane (3×20 mL). The combined organic phases are washed with brine (1×20 mL) and dried with magnesium sulfate. After evaporation of the solvent the crude product is purified by column chromatography (silica gel, Cy/EtOAc, 1:3). The silica gel was inactivated by flushing with the eluent containing 2% triethylamine. The product is obtained as highly viscose liquid, which on drying in high vacuum becomes solid.
According to the general procedure (S)—N-(4-methoxyphenylsulfonyl)-N-(3-pyridylmethyl)aminobutanoic acid (185 mg, 0.51 mmol) was reacted with HOBT (71 mg, 0.51 mmol), NMM (259 mg, 2.55 mmol), O-tert-butylhydroxylamine (194 mg, 1.53 mmol) and EDC (134 mg, 0.69 mmol) in dichloromethane (15 mL). After purification by column chromatography (column Ø 2 cm×15) the product was isolated as white solid. Yield: 173 mg (78%). M.p. 66° C. 1H NMR (300 MHz, DMSO-d6): δ 0.69 (t, 3JH,H=7.2 Hz, 3H, 4-CH3), 1.23 (s, 9H, 6-CH3), 1.44 (tq, 2JH,H=14.8 Hz, 3JH,H=7.4 Hz, 1H, 3-CHA), 1.90 (m, 1H, 3-CHB), 3.85 (s, 3H, 11-CH3), 4.11 (m, 1H, 2-CH), 4.56 (s, 2H, 12-CH2), 6.91 (d, 3JH,H=8.9 Hz, 2H, 8-CH), 7.20 (dd, 3JH,H=7.8 Hz, 3JH,H=4.9 Hz, 1H, 16-CH), 7.61 (d, 3JH,H=8.9 Hz, 2H, 9-CH), 7.71 (d, 3JH,H=7.8 Hz, 1H, 17-CH), 8.48 (d, 3JH,H=3.7 Hz, 1H, 15-CH), 8.54 (d, 4JH,H=1.5 Hz, 1H, 14-CH), 8.59 (s, 1H, 18-NH). 13C NMR (75 MHz, DMSO-d6): δ 10.6 (q, C-4), 22.9 (t, C-3), 26.2 (q, C-6), 45.7 (t, C-12), 55.6 (q, C-11), 59.5 (d, C-2), 82.2 (s, C-5), 114.4 (d, C-8), 123.2 (d, C-16), 129.2 (d, C-9), 131.3 (s, C-7), 132.9 (s, C-13), 136.5 (d, C-17), 148.8 (d, C-15), 149.9 (d, C-14), 163.2 (s, C-10), 168.2 (s, C-1). Exact mass (ESI+): C21H29N3O5S+H+, calcd. 436.1906. found 436.1898; C21H29N3O5S+Na+, calcd. 458.1726. found 458.1716; (C21H29N3O5S)2+H+, calcd. 871.3734. found 871.3717; (C21H29N3O5S)2+Na+, calcd. 893.3554. found 893.3534. MS (ESI+, daughter ion experiment): m/z (%)=436 (50) [M++H+], 380 (100) [436-C4H8], 319 (18) [436-H+—CONHOC(CH3)3], 264 (6) [436-H+—C7H7O3S], 228 (4) [319-C6H6N], 171 (12) [C7H7O3S+]. Optical rotation: [α]58920=−37.2, [α]57820=−39.3, [α]54620=−45.2, [α]43620=−84.7, [α]36520=−156.2 (c=1.007, CHCl3).
According to the general procedure (R)—N-(4-methoxyphenylsulfonyl)-N-(3-pyridylmethyl)-aminobutanoic acid (105 mg, 0.29 mmol) was reacted with HOBT (40 mg, 0.29 mmol), NMM (147 mg, 1.45 mmol), O-tert-butylhydroxylamine (110 mg, 0.87 mmol) and EDC (74 mg, 0.38 mmol) in dichloromethane (15 mL). After column chromatography (column Ø 2 cm×15) the product was isolated as white solid. Yield: 101 mg (83%). M.p. 65-66° C. 1H NMR (300.13 MHz, DMSO-d6): δ 0.69 (t, 3JH,H=7.3 Hz, 3H, 4-CH3), 1.25 (s, 9H, 6-CH3), 1.44 (m, 1H, 3-CHA), 1.90 (m, 1H, 3-CHB), 3.85 (s, 3H, 11-CH3), 4.09 (m, 1H, 2-CH), 4.56 (s, 2H, 12-CH2), 6.91 (dm, 3JH,H=8.9 Hz, 2H, 8-CH), 7.20 (dd, 3JH,H=7.8 Hz, 3JH,H=4.8 Hz, 1H, 16-CH), 7.61 (dm, 3JH,H=8.9 Hz, 2H, 9-CH), 7.70 (d, 3JH,H=7.8 Hz, 1H, 17-CH), 8.49 (d, 3JH,H=3.7 Hz, 1H, 15-CH), 8.53-8.56 (m, 2H, 14-CH and 18-NH). 13C NMR (75.48 MHz, DMSO-d6): δ 10.6 (q, C-4), 22.9 (t, C-3), 26.2 (q, C-6), 45.7 (t, C-12), 55.6 (q, C-11), 59.5 (d, C-2), 82.2 (s, C-5), 114.4 (d, C-8), 123.2 (d, C-16), 129.2 (d, C-9), 131.4 (s, C-7), 132.9 (s, C-13), 136.5 (d, C-17), 148.9 (d, C-15), 150.0 (d, C-14), 163.2 (s, C-10), 168.2 (s, C-1). Exact mass (ESI+): C21H29N3O5S)2+Na+, calcd. 436.1906. found 436.1897; C21H29N3O5S+Na+, calcd. 458.1726. found 458.1715; (C21H29N3O5S)2+H+, calcd. 871.3734. found 871.3702; (C21H29N3O5S)2+Na+, calcd. 893.3554. found 893.3521. MS (ESI+, daughter ion experiment): m/z (%) 436 (65) [M++H+], 380 (100) [436-C4H8], 319 (12) [436-H+-CONHOC(CH3)3], 264 (15) [436-H+—C7H7O3S], 228 (5) [319-C6H6N]. Optical rotation: [α]58920=+37.2, [α]57820=+39.1, [α]54620=+44.9, [α]43620=+84.3, [α]36520=+155.4 (c=1.004, CHCl3).
According to the general procedure (S)—N-(4-methoxyphenylsulfonyl)-N-(3-pyridylmethyl)-aminopent-4-enoic acid (280 mg, 0.744 mmol) was reacted with HOBT (101 mg, 0.744 mmol), NMM (377 mg, 3.72 mmol), O-tert-butylhydroxylamine (280 mg, 2.23 mmol) and EDC (186 mg, 0.97 mmol) in dichloromethane (15 mL). After column chromatography (column Ø 2 cm×15 cm) the product was isolated as white solid. Yield: 306 mg (92%). M.p. 65-66° C. 1H NMR (300 MHz, CDCl3): δ 1.21 (s, 9H, 7-CH3), 2.16 (dt, 2JH,H=13.8 Hz, 3JH,H=6.8 Hz, 1H, 3-CHA), 2.60 (ddd, 2JH,H=14.0 Hz, 3JH,H=8.1 Hz, 3JH,H=7.0 Hz, 1 H, 3-CHB), 3.86 (s, 3H, 12-CH3), 4.29 (dd, 3JH,H=8.3 Hz, 3JH,H=6.9 Hz, 1H, 2-CH), 4.58 (s, 2H, 13-CH2), 4.94 (d, 2JH,H=2.9 Hz, 1H, 5-CHA), 4.97 (d, 2JH,H=3.1 Hz, 1H, 5-CHB), 5.39 (ddt, 3JH,H=14.1 Hz, 3JH,H=9.6 Hz, 3JH,H=6.9 Hz, 1H, 4-CH), 6.93 (dm, 3JH,H=8.9 Hz, 2H, 9-CH), 7.21 (dd, 3JH,H=7.7 Hz, 3JH,H=4.9 Hz, 1H, 17-CH), 7.64 (dm, 3JH,H=9.0 Hz, 2H, 10-CH), 7.72 (d, 3JH,H=7.9 Hz, 1H, 18-CH), 8.49 (d, 3JH,H=3.7 Hz, 1H, 16-CH), 8.56 (d, 4JH,H=1.5 Hz, 1H, 15-CH), 8.70 (s, 1H, 19-NH). 13C NMR (75 MHz, CDCl3): δ 26.2 (q, C-7), 33.9 (t, C-3), 45.8 (t, C-13), 55.6 (q, C-12), 57.6 (d, C-2), 82.2 (s, C-6), 114.4 (d, C-9), 119.0 (t, C-5), 123.3 (d, C-17), 129.3 (d, C-10), 131.0 (s, C-8), 132.5 (d, C-4), 132.8 (s, C-14), 136.4 (d, C-18), 148.8 (d, C-16), 149.8 (d, C-15), 163.2 (s, C-11), 167.7 (s, C-1). Elemental analysis: C22H29N3O5S (M=447.18 g/mol), calcd. C, 59.04; H, 6.53; N, 9.39. found C, 59.11; H, 6.57; N, 9.10%. Exact mass (ESI+): C22H29N3O5S+H+, calcd. 448.1906. found 448.1907; C22H29N3O5S+Na+, calcd. 895.3734. found 895.3730; (C22H29N3O5S)2+H+, calcd. 470.1726. found 470.1722; (C22H29N3O5S)2+Na+, calcd. 917.3554. found 917.3552. MS (ESI+, daughter ion experiment): m/z (%) 448 (10) [M++H+], 392 (100) [448-C4H8], 331 (32) [448-C5H11NO2], 279 (8) [331-C4H6], 240 (7), 171 (30) [C7H7O3S+], 161 (20) [331-C7H7O3S], 125 (11), 107 (2) [C7H7O+], 92 (8) [C6H6N+]. Optical rotation: [α]58920=−30.4, [α]57820=−31.9, [α]54620=−37.0, [α]43620=−69.1, [α]36520=−126.5 (c=0.998, CHCl3).
According to the general procedure (R)—N-(4-methoxyphenylsulfonyl)-N-(3-pyridylmethyl)-aminopent-4-enoic acid (150 mg, 0.40 mmol) was reacted with HOBT (54 mg, 0.40 mmol), NMM (202 mg, 1.99 mmol), O-tert-butylhydroxylamine (151 mg 1.20 mmol) and EDC (100 mg 0.52 mmol) in dichloromethane (15 mL). After column chromatography (column Ø 2 cm×15 cm) the product was isolated as white solid. Yield: 130 mg (73%). M.p. 65-66° C. 1H NMR (300 MHz, CDCl3): δ 1.21 (s, 9H, 7-CH3), 2.17 (dt, 2JH,H=14.0 Hz, 3JH,H=6.9 Hz, 1H, 3-CHA), 2.60 (ddd, 2JH,H=14.8 Hz, 3JH,H=8.3 Hz, 3JH,H=6.8 Hz, 1H, 3-CHB), 3.86 (s, 3H, 12-CH3), 4.31 (t, 3JH,H=7.6 Hz, 1H, 2-CH), 4.60 (s, 2H, 13-CH2), 4.95 (dd, 3JH,H=13.7 Hz, 2JH,H=1.9 Hz, 2H, 5-CH2), 5.40 (td, 3JH,H=16.8 Hz, 3JH,H=6.9 Hz, 1H, 4-CH), 6.93 (dm, 3JH,H=8.9 Hz, 2H, 9-CH), 7.23 (dd, 3JH,H=7.8 Hz, 3JH,H=4.9 Hz, 1H, 17-CH), 7.65 (dm, 3JH,H=8.9 Hz, 2H, 10-CH), 7.75 (d, 3JH,H=7.9 Hz, 1H, 18-CH), 8.50 (d, 3JH,H=3.8 Hz, 1H, 16-CH), 8.58 (s, 1H, 15-CH), 8.75 (s, 1H, 19-NH). 13C NMR (75 MHz, CDCl3): δ 26.2 (q, C-7), 34.0 (t, C-3), 45.7 (t, C-13), 55.6 (q, C-12), 57.6 (d, C-2), 82.2 (s, C-6), 114.4 (d, C-9), 118.9 (t, C-5), 123.3 (d, C-17), 129.3 (d, C-10), 131.0 (s, C-8), 132.5 (d, C-4), 133.1 (s, C-14), 136.8 (d, C-18), 148.5 (d, C-16), 149.5 (d, C-15), 163.2 (s, C-11), 167.7 (s, C-1). Exact mass (ESI+): C22H29N3O5S+Fr, calcd. 448.1906. found 448.1900; C22H29N3O5S+Na+, calcd. 470.1726. found 470.1722; (C22H29N3O5S)2+H+, calcd. 895.3734. found 895.3730; (C22H29N3O5S)2+Na+, calcd. 917.3554. found 917.3546. MS (ESI+, daughter ion experiment): m/z (%) 448 (50) [M++H+], 392 (100) [448-C4H8], 331 (13) [448-H+-CONHOC(CH3)3], 240 (8) [331-C6H6N], 171 (12) [C7H7O3S+]. Optical rotation: [α]58920=+29.6, [α]57820=+31.2, [α]54620=+36.0, [α]43620=+66.9, [α]36520=+121.9 (c=1.010, CHCl3).
According to the general procedure (S)—N-(4-methoxyphenylsulfonyl)-N-(3-pyridylmethyl)-2-amino-4-fluorobutanoic acid (395 mg, 1.03 mmol) was reacted with HOBT (140 mg, 1.03 mmol), NMM (522 mg, 5.17 mmol), O-tert-butylhydroxylamine (390 mg, 3.10 mmol) and EDC (257 mg, 1.34 mmol) in dichloromethane (15 mL). After column chromatography (column Ø 2 cm×15 cm) the product was isolated as white solid. Yield: 355 mg (75%). M.p. 59-60° C. 1H NMR (300 MHz, CDCl3): δ 1.24 (s, 9H, 6-CH3), 1.72 (m, 3JH,F=22.5 Hz, 1H, 3-CHA), 2.22 (m, 3JH,F=31.6 Hz, 1H 3-CHB), 3.86 (s, 3H, 11-CH3), 4.25 (m, 2JH,F=47.6 Hz, 2H, 4-CH2F), 4.49 (d, 3JH,H=16.2 Hz, 1H, 12-CHA), 4.55 (m, 1H, 2-CH), 4.61 (d, 3JH,H=16.0 Hz, 1H, 12-CHB), 6.94 (d, 3JH,H=8.8 Hz, 2H, 8-CH), 7.22 (dd, 3JH,H=7.8 Hz, 3JH,H=4.8 Hz, 1H, 16-CH), 7.62-7.71 (m, 3JH,H=8.9 Hz, 3H, 9-CH & 17-CH), 8.49-8.53 (m, 2H, 14-CH & 15-CH), 8.81 (m, 1H, 18-NH). 13C NMR (75 MHz, CDCl3): δ 26.9 (s, C-6), 30.6 (d, 2JH,F=19.5 Hz, C-3), 45.8 (s, C-12), 53.9 (d, 3JH,F=3.0 Hz, C-2), 55.6 (s, C-11), 80.0 (d, 1JH,F=165.6 Hz, C-4), 82.4 (s, C-5), 114.5 (s, C-8), 123.4 (s, C-16), 129.2 (s, C-9), 130.8 (s, C-7), 132.6 (s, C-13), 136.4 (s, C-17), 149.1 (s, C-15), 149.8 (s, C-14), 163.3 (s, C-10), 167.4 (s, C-1). 19F NMR (282 MHz, CDCl3): δ −222.2 (ddt, 2JH,F=47.5 Hz, 3JH,F=31.5 Hz, 3JH,F=22.6 Hz, 4-CH2F). Exact mass (ESI+): C21H28FN3O5S+H+, calcd. 454.1812. found 454.1804; C21H28FN3O5S+Na+, calcd. 476.1631. found 476.1623; (C21H28FN3O5S)2+H+, calcd. 907.3546. found 907.3544; (C21H28FN3O5S)2+Na+, calcd. 929.3365. found 929.3370. MS (GC/MS, 70 eV): m/z (%) 454 (100) [M++H+], 398 (97) [M+-C4H8], 337 (15) [M+-CONHOC(CH3)3], 317 (4) [337-HF], 246 (3) [337-C6H6N], 171 (15) [C7H7O3S+].
Optical rotation: [α]58920=+17.9, [α]57820=+18.7, [α]54620=+21.7, [α]43620=+40.7, [α]36520=+75.1 (c=1.005, CHCl3).
According to the general procedure (R)—N-(4-methoxyphenylsulfonyl)-N-(3-pyridylmethyl)-2-amino-4-fluorobutanoic acid (325 mg, 0.85 mmol) with HOBT (115 mg, 0.85 mmol), NMM (430 mg, 4.25 mmol), O-tert-butylhydroxylamine (320 mg, 2.55 mmol) and EDC (213 mg, 1.11 mmol) in dichloromethane (15 mL). After column chromatography (column Ø 2 cm×10 cm) the product as white solid. Yield: 353 mg (91%). M.p. 60° C. 1H NMR (400 MHz, CDCl3): δ 1.24 (s, 9H, 6-CH3), 1.71 (m, 3JH,F=22.7 Hz, 1H, 3-CHA), 2.22 (m, 3JH,F=28.2 Hz, 1H, 3-CHB), 3.87 (s, 3H, 11-CH3), 4.17-4.37 (m, 2JH,F=46.9 Hz, 2H, 4-CH2F), 4.50 (d, 2JH,H=16.0 Hz, 1H, 12-CHA), 4.55 (dd, 3JH,H=8.8 Hz, 3JH,H=5.9 Hz, 1H, 2-CH), 4.61 (d, 2JH,H=16.0 Hz, 1H, 12-CHB), 6.95 (dm, 3JH,H=8.9 Hz, 2H, 8-CH), 7.24 (dd, 3JH,H=7.8 Hz, 3JH,H=4.8 Hz, 1H, 16-CH), 7.66 (dm, 3JH,H=8.9 Hz, 2H, 9-CH), 7.71 (d, 3JH,H=7.9 Hz, 1H, 17-CH), 8.51 (d, 3JH,H=4.9 Hz, 4JH,H=1.2 Hz, 1H, 15-CH), 8.53 (d, 4JH,H=1.7 Hz, 1H, 14-CH), 8.76 (s, 1H, 18-NH). 13C NMR (100 MHz, CDCl3): δ 26.2 (s, C-6), 30.6 (d, 2JH,F=19.9 Hz, C-3), 55.6 (s, C-11), 80.0 (d, 1JH,F=165.9 Hz, C-4), 45.8 (s, C-12), 53.9 (d, 3JH,F=2.5 Hz, C-2), 82.4 (s, C-5), 114.5 (s, C-8), 123.4 (s, C-16), 129.3 (s, C-9), 130.8 (s, C-7), 132.7 (s, C-13), 136.6 (s, C-17), 148.8 (s, C-15), 149.6 (s, C-14), 163.4 (s, C-10), 167.4 (s, C-1). 19F NMR (282 MHz, CDCl3): δ −221.4 (ddt, 2JH,F=46.9 Hz, 3JH,F=28.4 Hz, 3JH,F=22.8 Hz, 4-CH2F). Elemental analysis: C21H30FN3O5S (M=455.54 g/mol): calcd. C, 55.61, H, 6.22, N, 9.27%. found C, 55.24; H, 6.47; N, 9.22%. Exact mass (ESI+): C21H28FN3O5S+H+, calcd. 454.1812. found 454.1817; C21H28FN3O5S+Na+, calcd. 476.1631. found 476.1635; (C21H28FN3O5S)2+H+, calcd. 907.3546. found 907.3567; (C21H28FN3O5S)2+Na+, calcd. 929.3365. found 929.3387. MS (GC/MS, 70 eV): m/z (%) 453 (0) [M+], 433 (2) [M+—HF], 377 (10) [433-C4H8], 347 (13), 277 (4) [M+-C8H15FNO2], 262 (47) [277-CH3], 213 (3) [C9H11NO3S+], 206 (100), 171 (20) [C7H7O3S+], 155 (14), 107 (40) [C7H7O3S+], 92 (34) [C6H6N+], 77 (8) [C6H5+], 57 (15) [C4H9+], 56 (3) [C4H8+]. Optical rotation: [α]58920=−14.8, [α]57820=−15.7, [α]54620=+18.1, [α]43620=+34.3, [α]36520=−62.8 (c=1.004, CHCl3).
According to the general procedure (S)—N-(4-methoxyphenylsulfonyl)-N-(3-pyridylmethyl)-2-amino-4-fluoropent-4-enoic acid (150 mg, 0.38 mmol) with HOBT (52 mg, 0.38 mmol), NMM (193 mg, 1.90 mmol), O-tert-butylhydroxylamine (144 mg, 1.14 mmol) and EDC (95 mg, 0.50 mmol) in dichloromethane (15 mL). After column chromatography (column: Ø 2 cm×10 cm) was isolated as white solid. Yield: 107 mg (61%). M.p. 60-61° C. 1H NMR (300 MHz, CDCl3): δ 1.23 (s, 9H, 7-CH3), 2.26 (ddd, 3JH,F=17.5 Hz, 2JH,H=14.7 Hz, 3JH,H=6.2 Hz, 1H, 3-CHA), 2.80 (ddd, 3JH,F=19.6 Hz, 2JH,H=14.7 Hz, 3JH,H=8.2 Hz, 1H, 3-CHB), 3.86 (s, 3H, 12-CH3), 4.15 (dd, 3JH,F=49.7 Hz, 2JH,H=3.1 Hz, 1H, 5-CHtrans 4.48 (dd, 3JH,F=17.1 Hz, 2JH,H=3.1 Hz, 1H, 5-CHcis), 4.50 (d, 2JH,H=16.2 Hz, 1H, 13-CHA), 4.59 (m, 1H, 2-CH), 4.62 (d, 2JH,H=16.0 Hz, 1H, 13-CHB), 6.93 (d, 3JH,H=8.9 Hz, 2H, 9-CH), 7.20 (dd, 3, JH,H=7.8 Hz, 3JH,H=4.8 Hz, 1H, 17-CH), 7.67 (t, 3JH,H=8.9 Hz, 3H, 10-CH and 18-CH), 8.49 (d, 3JH,H=3.8 Hz, 1H, 16-CH), 8.53 (d, 4JH,H=1.7 Hz, 1H, 15-CH), 8.69 (s, 1H, 19-NH). 13C NMR (75.48 MHz, CDCl3): δ 26.2 (q, C-7), 32.8 (dt, 2JH,F=27.9 Hz, C-3), 45.9 (t, C-13), 54.9 (d, C-2), 55.7 (q, C-12), 82.5 (s, C-6), 93.9 (dt, 2JH,F=18.5 Hz, C-5), 114.4 (d, C-9), 123.3 (d, C-17), 129.5 (d, C-10), 130.9 (s, C-8), 132.5 (s, C-14), 136.4 (d, C-18), 149.1 (d, C-16), 145.0 (d, C-15), 161.1 (ds, 1JH,F=256.4 Hz, C-4), 163.4 (s, C-11), 166.9 (s, C-1). 19F NMR (282 MHz, CDCl3): δ −98.4 (ddd, 3JH,F=49.6 Hz, 3JH,F=17.9 Hz, 3JH,F=17.6 Hz, 4-CF). Exact mass (ESI+): C22H28FN3O5S+H+, calcd. 466.1812. found 466.1819; C22H28FN3O5S+Na+, calcd. 488.1631. found 488.1634; (C22H28FN3O5S)2+H+, calcd. 931.3546. found 931.3565; (C22H28FN3O5S)2+Na+, calcd. 953.3384. found 953.3384. MS (ESI+, daughter ion experiment): m/z (%) 466 (18) [M++H+], 410 (100) [466-C4H8], 349 (13) [466-H+-CONHOC(CH3)3], 179 (11) [410-C7H7O3S+—C3H4F], 171 (30) [C7H7O3S+], [α]58920=−6.5, [α]57820=−6.8, [α]54620=−7.9, [α]43620=−14.3, [α]36520=−24.9 (c=0.996, CHCl3).
According to the general procedure (R)—N-(4-methoxyphenylsulfonyl)-N-(3-pyridylmethyl)-2-amino-4-fluoropent-4-enoic acid (150 mg, 0.38 mmol) was reacted with HOBT (52 mg, 0.38 mmol), NMM (193 mg, 1.90 mmol), O-tert-butylhydroxylamine (144 mg, 1.14 mmol) and EDC (95 mg, 0.50 mmol) in dichloromethane (15 mL). After column chromatography (column: Ø 2 cm×10 cm) the product was isolated as white solid. Yield: 112 mg (63%). M.p. 59-60° C. 1H NMR (300 MHz, CDCl3): δ 1.24 (s, 9H, 7-CH3), 2.27 (m, 1H, 3-CHA), 2.79 (ddd, 3JH,F=19.8 Hz, 2JH,H=14.7 Hz, 3JH,H=8.3 Hz, 1H, 3-CHB), 3.86 (s, 3H, 12-CH3), 4.15 (dd, 3JH,F=49.8 Hz, 2JH,H=3.1 Hz, 1H, 5-CHtrans), 4.48 (dd, 3JH,F=17.0 Hz, 2JH,H=3.2 Hz, 1H, 5-CHcis), 4.50 (d, 2JH,H=16.1 Hz, 1H, 13-CHA), 4.56-4.66 (m, 2H, 2-CH and 13-CHB), 6.93 (d, 3JH,H=9.0 Hz, 2H, 9-CH), 7.21 (dd, 3JH,H=7.8 Hz, 3JH,H=4.9 Hz, 1 H, 17-CH), 7.62-7.73 (m, 3H, 10-CH & 18-CH), 8.50 (dd, 3JH,H=5.3 Hz, 2JH,H=1.3 Hz, 1 H, 16-CH), 8.53 (d, 2JH,H=1.3 Hz, 1H, 15-CH), 8.72 (s, 1H, 19-NH). 13C NMR (75 MHz, CDCl3): δ 26.2 (q, C-7), 32.8 (dt, 2JH,F=27.7 Hz, C-3), 45.8 (t, C-13), 54.8 (d, C-2), 55.7 (q, C-12), 82.5 (s, C-6), 94.0 (dt, 2JH,F=18.2 Hz, C-5), 114.4 (d, C-9), 123.3 (d, C-17), 129.5 (d, C-10), 130.8 (s, C-8), 132.5 (s, C-14), 136.5 (d, C-18), 149.0 (d, C-16), 149.9 (d, C-15), 161.0 (ds, 1JH,F=256.9 Hz, C-4), 163.4 (s, C-11), 166.9 (s, C-1). 19F NMR (282 MHz, CDCl3): δ −98.4 (ddd, 3JH,F=49.8 Hz, 3JH,F=19.1 Hz, 3JH,F=17.4 Hz, 4-CF). Exact mass (ESI+): C22H28FN3O5S+H+, calcd. 466.1812. found 466.1806; C22H28FN3O5S+Na+, calcd. 488.1631. found 488.1626; (C22H28FN3O5S)2+H+, calcd. 931.3546. found 931.3519; MS (ESI+, daughter ion experiment): m/z (%) 466 (40) [M++H+], 410 (100) [466-C4H8], 349 (10) [466-H+—CONHOC(CH3)3], 171 (5) [C7H7O3S+]. Optical rotation: [α]58920=+8.5, [α]57820=+8.7, [α]54620=+10.1, [α]43620=+18.2, [α]36520=+31.8 (c=1.023, CHCl3).
General Procedure for the Hydrolysis of Hydroxamates to Hydroxamic Acids
In a dried YOUNG-tube, the corresponding hydroxamate is dissolved in fresh trifluoroacetic acid (50 mL/mmol) under argon. The YOUNG-tube is flashed with argon and closed. The solution is stirred at 40° C. for 12 h. Then the reaction mixture is evaporated to dryness in vacuum. The residue is dissolved in chloroform (100 mL/mmol) and washed with aqueous citric acid (pH 4) (25 mL/mmol) and bicarbonate. The aqueous phase is extracted with chloroform and the combined organic phases are dried with magnesium sulfate. After evaporation of the solvent the product is isolated as viscos oil.
According to the general procedure tert-butyl (S)—N-(4-methoxyphenyl-sulfonyl)-N-(3-pyridylmethyl)-2-aminobutanhydroxamate (173 mg, 0.40 mmol) was hydrolyzed in trifluoroacetic acid (25 mL). After work up, the product was isolated as a cloudy, highly viscos liquid. After purification with preparative HPLC the product was 96% pure. Yield: 60 mg (40%). 1H NMR (300.13 MHz, CD3CN): δ 0.71 (t, 3JH,H=7.4 Hz, 3H, 4-CH3), 1.40 (m, 1H, 3-CHA), 1.66 (m, 1H, 3-CHB), 3.86 (s, 3H, 9-CH3), 4.18 (dd, 3JH,H=8.7 Hz, 3JH,H=6.6 Hz, 1H, 2-CH), 4.74 (d, 2JH,H=17.4 Hz, 1H, 10-CHA), 4.86 (d, 2JH,H=17.4 Hz, 1H, 10-CHB), 7.03 (d, 3JH,H=9.0 Hz, 2H, 6-CH), 7.71 (d, 3JH,H=9.1 Hz, 2H, 7-CH), 7.91 (dd, 3JH,H=8.1 Hz, 3JH,H=5.8 Hz, 1H, 14-CH), 8.50 (d, 3JH,H=8.1 Hz, 1H, 15-CH), 8.58 (d, 3JH,H=5.7 Hz, 1H, 13-CH), 8.77 (s, 1H, 12-CH). 13C NMR (75.48 MHz, CD3CN): 10.7 (q, C-4), 23.8 (t, C-3), 46.0 (t, C-10), 56.7 (q, C-9), 59.7 (d, C-2), 115.6 (d, C-6), 127.7 (d, C-14), 130.5 (d, C-7), 131.6 (s, C-5), 141.0 (d, C-12), 141.1 (s, C-11), 142.1 (d, C-13), 146.4 (d, C-15), 164.6 (s, C-9), 168.3 (s, C-1). Exact mass (ESI+): C17H21N3O5S+H+, calcd. 380.1280. found 380.1263; C17H21N3O5S+Na+, calcd. 402.1100. found 402.1082. MS (ESI+, daughter ion experiment): m/z (%) 380 (100) [M++H+], 319 (25) [M+-CONHOH], 228 (5) [319-C6H6N], 171 (30) [C7H7O3S+], 92 (10) [C6H6N+].
According to the general procedure tert-butyl (R)—N-(4-methoxyphenyl-sulfonyl)-N-(3-pyridylmethyl)-2-aminobutanhydroxamate (101 mg, 0.23 mmol) was hydrolyzed in trifluoroacetic acid (25 mL). After workup, the product was isolated as white, highly viscos liquid. HPLC showed 95% purity of the product. Yield: 38 mg (43%). 1H NMR (300 MHz, CD3CN): δ 0.72 (t, 3JH,H=7.4 Hz, 3H, 4-CH3), 1.40 (m, 1H, 3-CHA), 1.76 (m, 1H, 3-CHB), 3.85 (s, 3H, 9-CH3), 4.17 (dd, 3JH,H=8.7 Hz, 3JH,H=6.6 Hz, 1H, 2-CH), 4.75 (d, 2JH,H=17.4 Hz, 1H, 10-CHA), 4.83 (d, 2JH,H=17.3 Hz, 1H, 10-CHB), 7.01 (d, 3JH,H=9.0 Hz, 2H, 6-CH), 7.69 (d, 3JH,H=9.0 Hz, 2H, 7-CH), 7.88 (dd, 3JH,H=8.0 Hz, 3JH,H=5.8 Hz, 1H, 14-CH), 8.44 (d, 3JH,H=8.1 Hz, 1H, 15-CH), 8.59 (d, 3JH,H=5.5 Hz, 1H, 13-CH), 8.77 (s, 1H, 12-CH). 13C NMR (75 MHz, CD3CN): δ 10.6 (q, C-4), 23.7 (t, C-3), 46.9 (t, C-10), 56.6 (q, C-9), 59.7 (d, C-2), 115.6 (d, C-6), 127.5 (d, C-14), 130.4 (d, C-7), 131.7 (s, C-5), 140.8 (d, C-12), 141.1 (s, C-11), 142.4 (d, C-13), 146.0 (d, C-15), 164.6 (s, C-9), 168.2 (s, C-1). Exact mass (ESI+): C17H21N3O5S+Na+, calcd. 380.1280. found 380.1278; C17H21N3O5S+Na+, calcd. 402.1100. found 402.1094. MS (ESI+, daughter ion experiment): m/z (%) 759 (100) [2M++H+], 380 (18) [M++H+], 279 (13), 227 (13) [C10H13NO3S+], 155 (8) [C7H7O2S+].
According to the general procedure tert-butyl (S)—N-(4-methoxyphenyl-sulfonyl)-N-(3-pyridylmethyl)-2-aminopent-4-enhydroxamate (300 mg, 0.67 mmol) was hydrolyzed in trifluoroacetic acid (25 mL). After workup, the product was isolated as white, highly viscos liquid. HPLC showed 96% purity of the product. Yield: 77 mg (30%). 1H NMR (400 MHz, CD3CN): δ 2.14 (dt, 2JH,H=14.3 Hz, 3JH,H=6.8 Hz, 1H, 3-CHA), 2.45 (dt, 2JH,H=14.2 Hz, 3JH,H=7.7 Hz, 1H, 3-CHB), 3.84 (s, 3H, 10-CH3), 4.31 (t, 3JH,H=7.6 Hz, 1H, 2-CH), 4.61 (d, 2JH,H=16.8 Hz, 1H, 11-CHA), 4.73 (d, 2JH,H=16.8 Hz, 1H, 11-CHB), 4.92 (s, 1H, 5-CHA), 4.95 (d, 3JH,H=5.6 Hz, 1H, 5-CHB), 5.47 (ddt, 3JH,H=17.3 Hz, 3JH,H=10.4 Hz, 3JH,H=6.9 Hz, 1H, 4-CH), 7.00 (d, 3JH,H=8.8 Hz, 2H, 7-CH), 7.30 (dd, 3JH,H=7.6 Hz, 3JH,H=4.9 Hz, 1H, 15-CH), 7.70 (d, 3JH,H=8.9 Hz, 2H, 8-CH), 7.81 (d, 3JH,H=7.8 Hz, 1H, 16-CH), 8.41 (s, 1H, 14-CH), 8.53 (s, 1H, 13-CH). 13C NMR (100 MHz, CD3CN): δ 35.2 (t, C-3), 46.8 (t, C-11), 56.6 (q, C-10), 58.0 (d, C-2), 115.4 (d, C-7), 119.1 (t, C-5), 124.5 (d, C-15), 130.5 (d, C-8), 132.1 (d, C-4), 133.9 (s, C-12), 135.9 (s, C-6), 137.7 (d, C-16), 148.5 (d, C-14), 149.5 (d, C-13), 164.4 (s, C-9), 167.4 (s, C-1). Exat mass (ESI+): C18H21N3O5S+H+, calcd. 392.1280. found 392.1271; C18H21N3O5S+Na+, calcd. 414.1100. found 414.1094. MS (ESI+, daughter ion experiment): m/z (%) 392 (10) [M++H+], 331 (15) [M+-CONHOH], 279 (4) [C13H15N2O3S+], 240 (8) [331-C5H4N], 171 (100) [C7H7O3S+], 161 (30) [C10H13N2+], 92 (45) [C6H6N+].
According to the general procedure tert-butyl (R)—N-(4-methoxyphenyl-sulfonyl)-N-(3-pyridylmethyl)-2-aminopent-4-enhydroxamate (100 mg, 0.22 mmol) was hydrolyzed in trifluoroacetic acid (25 mL). After workup, the product was isolated as white, highly viscos liquid. HPLC showed 97% purity of the product. Yield: 33 mg (38%). 1H NMR (400 MHz, CD3CN): δ 2.15 (dt, 2JH,H=14.1 Hz, 3JH,H=6.9 Hz, 1H, 3-CHA), 2.46 (dt, 2JH,H=14.1 Hz, 3, JH,H=7.6 Hz, 1H, 3-CHB), 3.84 (s, 3H, 10-CH3), 4.29 (t, 3JH,H=7.6 Hz, 1H, 2-CH), 4.60 (d, 2JH,H=16.7 Hz, 1H, 11-CHA), 4.72 (d, 2JH,H=16.7 Hz, 1H, 11-CHB), 4.93 (s, 1H, 5-CHA), 4.96 (d, 3JH,H=4.5 Hz, 1H, 5-CHB), 5.48 (ddt, 3JH,H=16.8 Hz, 3JH,H=9.6 Hz, 3JH,H=6.9 Hz, 1H, 4-CH), 7.00 (d, 3JH,H=8.7 Hz, 2H, 7-CH), 7.29 (dd, 3JH,H=7.8 Hz, 3JH,H=4.8 Hz, 1H, 15-CH), 7.70 (d, 3JH,H=8.9 Hz, 2H, 8-CH), 7.79 (m, 1H, 16-CH), 8.41 (d, 3JH,H=4.4 Hz, 1H, 14-CH), 8.52 (s, 1H, 13-CH). 13C NMR (100 MHz, CD3CN): 35.2 (t, C-3), 46.8 (t, C-11), 56.6 (q, C-10), 58.0 (d, C-2), 115.4 (d, C-7), 119.1 (t, C-5), 124.5 (d, C-15), 130.5 (d, C-8), 132.1 (d, C-4), 134.0 (s, C-12), 135.8 (s, C-6), 137.5 (d, C-16), 148.6 (d, C-14), 149.7 (d, C-13), 164.4 (s, C-9), 167.3 (s, C-1). Exact mass (ESI+): C18H21N3O5S+H+, calcd. 392.1280. found 392.1275. MS (ESI+, daughter ion experiment): m/z (%) 392 (30) [M+ +H+], 331 (35) [M+-CONHOH], 279 (8) [C13H15N2O3S+], 240 (20) [331-C5H4N], 171 (100) [C7H7O3S+], 161 (55) [C10H13N2+], 124 (27) [C7H8O2+], 107 (11) [C7H7O+], 92 (42) [C6H6N+].
According to the general procedure tert-butyl (S)—N-(4-methoxyphenyl-sulfonyl)-N-(3-pyridylmethyl)-2-amino-4-fluorobutanhydroxamate (131 mg, 0.29 mmol) was hydrolyzed in trifluoroacetic acid (25 mL). After workup, the product was isolated as white, highly viscos liquid. Purification by preparative HPLC delivered a product of 95% purity. Yield: 49 mg (42%). 1H NMR (600 MHz, CD3CN): δ 1.76 (m, 3JH,F=23.7 Hz, 1H, 3-CHA), 2.14 (ddd, 3JH,F=29.1 Hz, 2JH,H=11.8 Hz, 3JH,H=6.1 Hz, 1H, 3-CHB), 3.85 (s, 3H, 9-CH3), 4.20-4.42 (m, 2H, 4-CH2F), 4.48 (dd, 3JH,H=8.9 Hz, 3JH,H=6.1 Hz, 1H, 2-CH), 4.76 (d, 2JH,H=17.1 Hz, 1H, 10-CHA), 4.81 (d, 2JH,H=17.1 Hz, 1H, 10-CHB), 7.01 (d, 3JH,H=8.9 Hz, 2H, 6-CH), 7.68 (d, 3JH,H=8.9 Hz, 2H, 7-CH), 7.88 (dd, 3JH,H=8.1 Hz, 3JH,H=5.7 Hz, 1H, 14-CH), 8.43 (d, 3JH,H=7.7 Hz, 1H, 15-CH), 8.58 (d, 3JH,H=5.3 Hz, 1H, 12-CH), 8.74 (s, 1H, 13-CH). 13C NMR (150 MHz, CD3CN): δ 31.3 (dt, 2JH,F=20.0 Hz, C-3), 46.1 (t, C-10), 55.0 (dd, 3JH,F=3.6 Hz, C-2), 56.78 (q, C-9), 81.4 (dt, 1JH,F=164.2 Hz, C-4), 115.7 (d, C-6), 127.6 (d, C-14), 130.2 (d, C-7), 131.4 (s, C-5), 140.4 (s, C-11), 141.2 (d, C-12), 142.3 (d, C-13), 146.1 (d, C-15), 164.7 (s, C-8), 167.3 (s, C-1). 19F NMR (564 MHz, CD3CN): δ −221.4 (tdd, 2JH,F=47.1 Hz, 3JH,F=29.0 Hz, 3JH,F=23.7 Hz, 4-CH2F). Exact mass (ESI+): C17H20FN3O5S++H+, calcd. 398.1186. found 398.1181. C17H20FN3O5S++Na+, calcd. 420.1105. found 420.0999; (C17H20FN3O5S+)2+H+, calcd. 795.2294. found 795.2300; (C17H20FN3O5S+)2+Na+, calcd. 817.2113. found 817.2116. MS (ESI+, daughter ion experiment): m/z (%) 398 (10) [M++H+], 337 (16) [M+-CONHOH], 171 (100) [C7H7O3S+], 167 (30) [337-C7H7O3S], 135 (16) [C8H11N2+], 123 (22) [C7H7O2+], 107 (5) [C7H7O+], 92 (52) [C6H6N+].
According to the general procedure tert-butyl (R)—N-(4-methoxyphenyl-sulfonyl)-N-(3-pyridylmethyl)-2-amino-4-fluorobutanhydroxamate (104 mg, 0.23 mmol) was hydrolyzed in trifluoroacetic acid (25 mL). After workup, the product was isolated as white, highly viscos liquid. Purification by preparative HPLC delivered a product of 95% purity. Yield: 45 mg (49%). 1H NMR (400 MHz, CD3CN): δ 1.67 (m, 1H, 3-CHA), 2.13 (m, 1H, 3-CHB), 3.86 (s, 3H, 9-CH3), 4.17-4.46 (m, 2H, 4-CH2F), 4.49 (dd, 3JH,H=8.8 Hz, 3JH,H=6.1 Hz, 1H, 2-CH), 4.75 (d, 2JH,H=17.3 Hz, 1H, 10-CHA), 4.86 (d, 2JH,H=17.3 Hz, 1H, 10-CHB), 7.04 (d, 3JH,H=9.0 Hz, 2H, 6-CH), 7.71 (d, 3JH,H=9.0 Hz, 2H, 7-CH), 7.92 (dd, 3JH,H=8.0 Hz, 3JH,H=5.9 Hz, 1H, 14-CH), 8.50 (d, 3JH,H=8.2 Hz, 1H, 15-CH), 8.59 (d, 3JH,H=5.6 Hz, 1H, 13-CH), 8.75 (s, 1H, 12-CH). 13C NMR (100 MHz, CD3CN): δ [ppm] 31.5 (dt, 2JH,F=20.1 Hz, C-3), 46.2 (t, C-10), 55.0 (dd, 3JH,F=3.4 Hz, C-2), 56.7 (q, C-9), 81.3 (dt, 1JH,F=164.2 Hz, C-4), 115.7 (d, C-6), 127.9 (d, C-14), 130.6 (d, C-7), 131.3 (s, C-5), 140.8 (s, C-11), 141.2 (s, C-13), 142.1 (s, C-12), 146.7 (s, C-15), 164.8 (s, C-8), 167.3 (s, C-1). 19F NMR (282 MHz, CD3CN): δ [ppm]-222.2 (tdd, 2JH,F=47.2 Hz, 3JH,F=31.5 Hz, 3JH,F=22.3 Hz, 4-CH2F). Exact mass (ESI+): C17H20N3O5S+Fr, calcd. 398.1186. found 398.1171; C17H20N3O5S+Na+, calcd. 420.1005. found 420.0990; (C17H20N3O5S)2+H+, calcd. 795.2294. found 871.3691; (C17H20N3O5S)2+Na+, calcd. 817.2113. found 893.3507. MS (ESI+, daughter ion experiment): m/z (%) 398 (60) [M++H+], 337 (100) [M+-CONHOH], 317 (7) [337-HF], 246 (6), 171 (20) [C7H7O3S+].
According to the general procedure tert-butyl (S)—N-(4-methoxyphenyl-sulfonyl)-N-(3-pyridylmethyl)-2-amino-4-fluoropent-4-enhydroxamate (107 mg, 0.23 mmol) was hydrolyzed in trifluoroacetic acid (25 mL). After workup, the product was isolated as white, highly viscos liquid. Purification by preparative HPLC delivered a product of 97% purity. Yield: 62 mg (66%). 1H NMR (400 MHz, CD3CN): δ 2.35 (ddd, 3JH,F=18.1 Hz, 2JH,H=14.9 Hz, 3JH,H=6.6 Hz, 1H, 3-CHA), 2.74 (ddd, 3JH,F=18.9 Hz, 2JH,H=14.8 Hz, 3JH,H=8.4 Hz, 1H, 3-CHB), 3.85 (s, 3H, 10-CH3), 4.34 (dd, 3JH,F=50.3 Hz, 2JH,H=3.3 Hz, 1H, 5-CHtrans), 4.56 (dd, 3JH,F=17.3 Hz, 2JH,H=3.3 Hz, 1H, 5-CHcis), 4.57 (dd, 3JH,H=8.2 Hz, 3JH,H=6.5 Hz, 1H, 2-CH), 4.81 (s, 2H, 11-CH2), 7.01 (d, 3JH,H=9.0 Hz, 2H, 7-CH), 7.66 (d, 3JH,H=9.0 Hz, 2H, 8-CH), 7.86 (dd, 3JH,H=8.0 Hz, 3JH,H=5.8 Hz, 1H, 15-CH), 8.41 (d, 3JH,H=8.2 Hz, 1H, 16-CH), 8.59 (d, 3JH,H=5.5 Hz, 1H, 14-CH), 8.73 (s, 1H, 13-CH). 13C NMR (100 MHz, CD3CN): δ 33.2 (dt, 2JH,F=28.0 Hz, C-3), 46.1 (t, C-11), 55.5 (d, C-2), 56.7 (q, C-10), 94.5 (dt, 2JH,F=18.7 Hz, C-5), 115.7 (d, C-7), 127.5 (d, C-15), 130.5 (d, C-8), 131.5 (s, C-6), 140.1 (s, C-12), 141.4 (d, C-14), 142.6 (d, C-13), 145.7 (d, C-16), 162.5 (ds, 1JH,F=255.6 Hz, C-4), 164.7 (s, C-9), 166.8 (s, C-1). 19F NMR (282 MHz, CD3CN): δ −97.3 (ddd, 3JH,F=50.2 Hz, 3JH,F=18.2 Hz, 3JH,F=17.7 Hz, 4-CF). Exact mass (ESI+): C18H20FN3O5S++H+, calcd. 410.1186. found 410.1174; (C18H20FN3O5S+)2+H+, caldc. 819.2294. found 819.2278; (C18H20FN3O5S+)2+Na+, calcd. 841.2113. found 841.2096. MS (ESI+, daughter ion experiment): m/z (%) 410 (5) [M++H+], 349 (11) [M+-CONHOH], 179 (23) [349-C7H7O3S−], 171 (100) [C7H7O3S+], 135 (18) [C8H11N2+], 123 (20) [C7H7O2], 92 (50) [C6H6N+].
According to the general procedure tert-butyl (R)—N-(4-methoxyphenylsulfonyl)-N-(3-pyridylmethyl)-2-amino-4-fluoropent-4-enhydroxamate (112 mg, 0.24 mmol) was hydrolyzed in trifluoroacetic acid (25 mL). After workup, the product was isolated as white, highly viscos liquid. Purification by preparative HPLC delivered a product of 97% purity. Yield: 57 mg (58%): 1H NMR (300 MHz, CD3CN): δ 2.35 (ddd, 3JH,F=18.1 Hz, 2JH,H=14.9 Hz, 3JH,H=6.7 Hz, 1H, 3-CHA), 2.74 (ddd, 3JH,F=18.6 Hz, 2JH,H=14.8 Hz, 3JH,H=8.3 Hz, 1H, 3-CHB), 3.86 (s, 3H, 10-CH3), 4.34 (dd, 3JH,F=50.3 Hz, 2JH,H=3.2 Hz, 1H, 5-CHtrans), 4.56 (dd, 3JH,F=17.3 Hz, 2JH,H=3.2 Hz, 1H, 5-CHcis), 4.59 (t, 3JH,H=7.5 Hz, 1H, 2-CH), 4.79 (d, 2JH,H=17.5 Hz, 1H, 11-CHA), 4.86 (d, 2JH,H=17.5 Hz, 1H, 11-CHB), 7.01 (d, 3JH,H=9.0 Hz, 2H, 7-CH), 7.68 (d, 3JH,H=9.0 Hz, 2H, 8-CH), 7.89 (dd, 3JH,H=8.0 Hz, 3JH,H=5.8 Hz, 1H, 15-CH), 8.45 (d, 3JH,H=8.1 Hz, 1H, 16-CH), 8.59 (d, 3JH,H=5.5 Hz, 1H, 14-CH), 8.74 (s, 1H, 13-CH). 13C NMR (75 MHz, CD3CN): δ 33.2 (dt, 2JH,F=27.9 Hz, C-3), 46.1 (t, C-11), 55.4 (d, C-2), 56.7 (q, C-10), 94.5 (dt, 2JH,F=18.6 Hz, C-5), 115.7 (d, C-7), 127.6 (d, C-15), 130.6 (d, C-8), 131.3 (s, C-6), 140.4 (s, C-12), 141.2 (d, C-14), 142.2 (d, C-13), 146.1 (d, C-16), 162.4 (ds, 1JH,F=255.5 Hz, C-4), 164.7 (s, C-9), 166.8 (s, C-1). 19F NMR (282 MHz, CD3CN): δ −97.3 (ddd, 3JH,F=50.3 Hz, 3JH,F=18.2 Hz, 3JH,F=17.5 Hz, 4-CF). Exact mass (ESI+): C18H20FN3O5S++H+, calcd. 410.1186. found 410.1174; C18H20FN3O5S++Na+, calcd. 432.1005. found 432.0991; (C18H20FN3O5S+)2+H+: calcd. 819.2294. found 819.2278; (C18H20FN3O5S+)2+Na+, calcd. 841.2113. found 841.2103. MS (ESI+, daughter ion experiment): m/z (%) −410 (6) [M++H+], 349 (10) [M+-CONHOH], 179 (22) [349-C7H7O3S+], 171 (100) [C7H7O3S+], 135 (18) [C8H11N2+], 123 (22) [C7H7O2+], 92 (50) [C6H6N+].
General Procedure for Substitution with p-(2-halogenethoxy)-phenylsulfonyl chloride
The tert-butyl 4-fluoro-2-aminopent-4-enecarboxylate is dissolved in pyridine and treated with 1 equivalent of p-(2-halogenethoxy)phenylsulfonyl chloride under stiffing at 0° C. The reaction mixture is allowed to warm up to r.t. and stirred for 40 h. Then the mixture is diluted with dichloromethane and the organic solution is washed with 0.5 N HCl, water (2 times each) and saturated aqueous sodium chloride. After drying with magnesium sulfate the solvent was removed in vacuum. The product is passed through a short pad of silica gel and recrystallized from ethyl acetate/cyclohexane. The products are isolated as white crystalline solids.
Prepared from tert-butyl (R)-2-amino-4-fluoropent-4-enecarboxylate (1000 mg, 5.28 mmol) and p-(2-fluorethoxy)phenylsulfonyl chloride (1225 mg, 5.28 mmol). Yield: 1.32 g (64%). M.p. 102° C. (EtOAc/Cy). 1H NMR (300 MHz, CDCl3): δ=7.79 (dm, 3JH,H=9.0 Hz, 2H, 8-CH), 6.99 (dm, 3JH,H=9.0 Hz, 2H, 7-CH), 5.37 (d, 3JH,H=8.9 Hz, 1H, 14-NH), 4.78 (dm, 2JH,F=47.3 Hz, 2H, 11-CH2), 4.64 (dd, 3JH,F=17.0 Hz, 2JH,H=3.1 Hz, 1H, 5-CHcis), 4.35 (dd, 3JH,F=49.3 Hz, 2JH,H=3.1 Hz, 1H, 5-CHtrans), 4.26 (dm, 3JH,F=27.7 Hz, 2H, 10-CH2), 3.98 (dt, 3JH,H=9.0 Hz, 3JH,H=5.9 Hz, 1H, 2-CH), 2.65 (d, 3JH,H=5.9 Hz, 1H, 3-CHA), 2.59 (dd, 3JH,H=5.7 Hz, 3JH,H=4.2 Hz, 1H, 3-CHB), 1.30 (s, 9H, 13-CH3). 13C NMR (75 MHz, CDCl3): δ=169.2 (s, C-1), 161.8 (s, C-9), 160.9 (ds, 1JH,F=257.1 Hz, C-4), 131.9 (s, C-6), 129.5 (d, C-8), 114.7 (d, C-7), 94.1 (dt, 2JH,F=18.8 Hz, C-5), 83.2 (s, C-12), 81.5 (dt, 1JH,F=171.5 Hz, C-11), 67.4 (dt, 2JH,F=20.4 Hz, C-10), 53.2 (d, C-2), 36.4 (dt, 2JH,F=27.4 Hz, C-3), 27.6 (q, C-13). 19F NMR (282 MHz, CDCl3): δ =−96.1 (ddt, 3JH,F=49.3 Hz, 3JH,F=20.4 Hz, 3JH,F=17.2 Hz, 4-CF), −224.3 (tt, 2JH,F=47.3 Hz, 3JH,F=27.8 Hz, 11-CH2F). Elemental analysis: C17H23F2NO5S (M=391.43 g/mol), alcld. C, 52.16; H, 5.92; N, 3.58. found C, 52.64; H, 6.24; N, 3.55%. Exact mass (ESI+): C17H23F2NO5S+Na+: cald. 414.1163. found 414.1160. (C17H23F2NO5S)2+Na+: cald. 805.2428. found 805.2423. Optical rotation: [α]58920=−4.0, [α]57820=−4.4, [α]54620=−5.3, [α]43620=−13.0, [α]36520=−31.1 (c=1.023, CHCl3)
Prepared from tert-butyl (S)-2-amino-4-fluoropent-4-enecarboxylate (892 mg, 4.7 mmol) with p-(2-fluoroethoxy)-phenylsulfonylchloride (1130 mg, 4.7 mmol). Yield: 1.18 g (64%). M.p. 102° C. (EtOAc/Cy). 1H NMR (300 MHz, CDCl3): δ=7.79 (dm, 3JH,H=9.0 Hz, 2H, 8-CH), 6.99 (dm, 3JH,H=9.0 Hz, 2H, 7-CH), 5.37 (d, 3JH,H=9.0 Hz, 1H, 14-NH), 4.77 (dm, 2JH,F=47.5 Hz, 2H, 11-CH2), 4.64 (dd, 3JH,F=17.1 Hz, 2JH,H=3.1 Hz, 1H, 5-CHcis), 4.35 (dd, 3JH,F=49.3 Hz, 2JH,H=3.1 Hz, 1H, 5-CHtrans), 4.26 (dm, 3JH,F=27.7 Hz, 2H, 10-CH2), 3.98 (dt, 3JH,H=8.8 Hz, 3JH,H=5.8 Hz, 1H, 2-CH), 2.65 (d, 3JH,H=5.9 Hz, 1H, 3-CHA), 2.59 (dd, 3JH,H=5.7 Hz, 3JH,H=4.2 Hz, 1H, 3-CHB), 1.30 (s, 9H, 13-CH3). 13C NMR (75 MHz, CDCl3): δ=169.2 (s, C-1), 161.8 (s, C-9), 160.9 (ds, 1JH,F=256.9 Hz, C-4), 131.9 (s, C-6), 129.5 (d, C-8), 114.7 (d, C-7), 94.1 (dt, 2JH,F=18.8 Hz, C-5), 83.2 (s, C-12), 81.5 (dt, 1JH,F=171.5 Hz, C-11), 67.4 (dt, 2JH,F=20.4 Hz, C-10), 53.2 (d, 3JC,F=1.0 Hz, C-2), 36.4 (dt, 2JC,F=27.6 Hz, C-3), 27.6 (q, C-13). 19F NMR (282 MHz, CDCl3): δ =−96.1 (ddt, 3JH,F=49.3 Hz, 3JH,F=20.4 Hz, 3JH,F=17.2 Hz, 4-CF), −224.3 (tt, 2JH,F=47.3 Hz, 3JH,F=27.7 Hz, 11-CH2F). Elemental analysis: C17H23F2NO5S (M=391.43 g/mol). Calcd. C, 52.16; H, 5.92; N, 3.58. found C, 52.19; H, 6.09; N, 3.54%. Optical rotation: [α]58920=−4.0, [α]57820=−4.4, [α]54620=−5.3, [α]43620=−13.0, [α]36520=−31.1 (c=1.005, CHCl3).
Prepared from tert-butyl (R)-2-amino-4-fluoropent-4-enoate (1000 mg, 5.28 mmol) and p-(2-chlorethoxy)-phenylsulfonylchloride (1350 mg, 5.28 mmol). Yield: 1.29 g (61%). M.p. 98-99° C. (EtOAc/Cy). 1H NMR (300 MHz, CDCl3): δ=7.80 (dm, 3JH,H=9.0 Hz, 2H, 8-CH), 6.98 (dm, 3JH,H=9.0 Hz, 2H, 7-CH), 5.29 (d, 3JH,H=9.0 Hz, 1H, 14-NH), 4.65 (dd, 3JH,F=17.0 Hz, 2JH,H=3.1 Hz, 1H, 5-CHcis), 4.35 (q, 3JH,F=49.5 Hz, 2JH,H=2.9 Hz, 2H, 5-CHtrans), 4.27 (t, 3JH,H=5.7 Hz, 2H, 10-CH2), 3.98 (dt, 3JH,H=8.9 Hz, 3JH,H=5.7 Hz, 1H, 2-CH), 3.84 (t, 3JH,H=5.7 Hz, 2H, 11-CH2Cl), 2.65 (dd, 3JH,H=5.8 Hz, 2JH,H=0.9 Hz, 1H, 3-CHA), 2.59 (t, 3JH,H=5.3 Hz, 1H, 3-CHB), 1.30 (s, 9H, 13-CH3). 13C NMR (75 MHz, CDCl3): δ=169.2 (s, C-1), 161.6 (s, C-9), 160.9 (ds, 1JH,F=257.1 Hz, C-4), 132.0 (s, C-6), 129.5 (d, C-8), 114.7 (d, C-7), 94.2 (dt, 2JH,F=18.9 Hz, C-5), 83.2 (s, C-12), 68.2 (t, C-10), 53.2 (dd, 3JH,F=0.9 Hz, C-2), 41.5 (t, C-11), 36.5 (dt, 2JH,F=27.2 Hz, C-3), 27.7 (q, C-13). 19F NMR (282 MHz, CDCl3): δ =−96.1 (ddt, 3JH,F=49.2 Hz, 3JH,F=20.6 Hz, 3JH,F=17.1 Hz, 4-CF). Elemental analysis: C17H23ClFNO5S (M=407.88 g/mol); calcd. C, 50.06; H, 5.68; N, 3.43. found C, 50.02, H, 5.51N, 3.36%. Exact mass (ESI+): C17H23ClFNO5S+Na+; calcd. 430.0867. found 430.0851; (C17H23ClFNO5S)2+Na+; calcd. 837.1837. found 837.1824. Optical rotation: [α]58920=+6.3, [α]57820=+6.9, [α]54620=+8.1, [α]43620=+18.5, [α]36520=+42.5 (c=0.991, CHCl3).
Prepared from tert-butyl (S)-2-amino-4-fluoropent-4-enoate (1000 mg, 5.28 mmol) and p-(2-chlorethoxy)phenylsulfonylchloride (1350 mg 5.28 mmol). Yield: 1.58 g (73%). 1H NMR (300 MHz, CDCl3): δ=7.80 (dm, 3JH,H=9.0 Hz, 2H, 8-CH), 6.98 (dm, 3JH,H=9.0 Hz, 2H, 7-CH), 5.32 (d, 3JH,H=8.9 Hz, 1H, 14-NH), 4.64 (dd, 3JH,F=17.1 Hz, 2JH,H=3.0 Hz, 1H, 5-CHcis), 4.35 (q, 3JH,F=49.4 Hz, 2JH,H=3.0 Hz, 2H, 5-CHtrans), 4.27 (t, 3JH,H=5.7 Hz, 2H, 10-CH2), 3.98 (dt, 3JH,H=9.0 Hz, 3JH,H=5.8 Hz, 1H, 2-CH), 3.84 (t, 3JH,H=5.7 Hz, 2H, 11-CH2Cl), 2.65 (dd, 3JH,H=5.9 Hz, 1H, 3-CHA), 2.59 (m, 3JH,H=5.4 Hz, 1H, 3-CHB), 1.30 (s, 9H, 13-CH3). 13C NMR (75 MHz, CDCl3): δ=169.2 (s, C-1), 161.6 (s, C-9), 160.9 (ds, 1JH,F=257.1 Hz, C-4), 132.0 (s, C-6), 129.5 (d, C-8), 114.7 (d, C-7), 94.2 (dt, 2JH,F=18.8 Hz, C-5), 83.2 (s, C-12), 68.2 (t, C-10), 53.2 (dd, 3JH,F=0.9 Hz, C-2), 41.5 (t, C-11), 36.5 (dt, 2JH,F=27.6 Hz, C-3), 27.6 (q, C-13). 19F NMR (282 MHz, CDCl3): δ =−96.1 (ddt, 3JH,F=49.4 Hz, 3JH,F=20.5 Hz, 3JH,F=17.1 Hz, 1H). Elemental analysis: C17H23ClFNO5S (M=407.88 g/mol), calcd. C, 50.06; H, 5.68; N, 3.43. found C, 49.90; H, 5.53; N, 3.38%. Exact mass (ESI+): C17H23ClFNO5S+Na+, calcd. 430.0867. found 430.0856; (C17H23ClFNO5S)2+Na+, calcd. 837.1837. found 837.1835. Optical rotation: [α]58920=−2.6, [α]57820=−2.9, [α]54620=−3.5, [α]43620=−8.5, [α]36520=−19.8 (c=1.021, CHCl3).
General Procedure for Substitution with Benzyl Bromide
The N-substituted tert-butyl 2-amino-4-fluoropent-4-enoate dissolved in dimethylformamide is treated under stirring with 10 equivalents of potassium carbonate. After 20 min 1 equivalent of benzyl bromide is added and the mixture is stirred at r.t. for 40 h. Then water is added and the mixture is extracted with ethyl acetate (4 times). The combined organic extracts are washed with water (4 times), saturated sodium chloride solution and dried with magnesium sulfate. After removal of the solvent the crude product is purified chromatographically an (silica gel, Cy/EtOAc, 4:1). The products are obtained as colorless or yellowish viscos oils.
Prepared from tert-butyl N-[p-(2-fluoroethoxy)phenylsulfonyl]aminopent-4-enoate (783 mg, 2 mmol), benzyl bromide (342 mg, 2 mmol) and potassium carbonate 2.8 g (20 mmol). Yield: 600 mg (63%). 1H NMR (300 MHz, CDCl3): δ=7.82 (dm, 3JH,H=9.0 Hz, 2H, 8-CH), 7.29 (m, 5H, 14-CH, 15-CH, 16-CH), 6.98 (dm, 3JH,H=9.0 Hz, 2H, 7-CH), 4.80 (dm, 2JH,F=47.1 Hz, 2H, 11-CH2), 4.66 (d, 2JH,H=16.3 Hz, 1H, 12-CHA), 4.56 (t, 3JH,H=7.3 Hz, 1H, 2-CH), 4.53 (dd, 3JH,F=17.1 Hz, 2JH,H=2.9 Hz, 1H, 5-CHcis), 4.29 (d, 2JH,H=15.9 Hz, 2H, 12-CHB), 4.27 (dm, 3JH,F=27.8 Hz, 2H, 10-CH2), 4.11 (dd, 3JH,F=49.8 Hz, 2JH,H=3.1 Hz, 1H, 5-CHtrans), 2.70 (dt, 3JH,F=15.4 Hz, 3JH,H=6.9 Hz, 1H, 3-CHA), 2.46 (ddd, 3JH,F=20.0 Hz, 2JH,H=15.1 Hz, 3JH,H=7.6 Hz, 1H, 3-CHB), 1.36 (s, 9H, 18-CH3). 13C NMR (75 MHz, CDCl3): δ=168.5 (s, C-1), 161.9 (ds, 1JH,F=256.5 Hz, C-4), 161.7 (s, C-9), 136.7 (s, C-13), 132.5 (s, C-6), 129.9 (d, C-8), 128.5 (d, C-14 o. C-15), 128.4 (d, C-14 o. C-15), 127.7 (d, C-16), 114.6 (d, C-7), 93.1 (dt, 2JH,F=19.1 Hz, C-5), 82.6 (s, C-17), 81.5 (dt, 1JH,F=171.7 Hz, C-11), 67.4 (dt, 2JH,F=20.5 Hz, C-10), 57.6 (d, C-2), 50.0 (t, C-12), 34.1 (dt, 2JH,F=27.9 Hz, C-3), 27.8 (q, C-18). 19F NMR (282 MHz, CDCl3): δ =−97.7 (m, 3JH,F=49.8 Hz, 3JH,F=20.0 Hz, 3JH,F=17.0 Hz, 4-CF), −224.3 (tt, 2JH,F=47.3 Hz, 3JH,F=27.6 Hz, 11-CH2F). Exact mass (ESI+): C24H29F2NO5S+Na+, calcd. 504.1632. found 504.1615; (C24H29F2NO5S)2+Na+, calcd. 985.3367. found 985.3348. Optical rotation: [α]58920=−22.8, [α]57820=−23.8, [α]54620=−27.2, [α]43620=−49.6, [α]36520=−80.6 (c=1.018 CHCl3).
Prepared from tert-butyl N-[p-(-2-fluoroethoxy)phenylsulfonyl]aminopent-4-enoate (830 mg, 2.12 mmol), benzyl bromide (363 mg, 2.12 mmol) and 2.93 g (21.2 mmol) potassium carbonate. Yield: 800 mg (1.7 mmol, 78%). 1H NMR (300 MHz, CDCl3): δ=7.82 (dm, 3JH,H=9.0 Hz, 2H, 8-CH), 7.29 (m, 5H, 14-CH, 15-CH, 16-CH), 6.98 (dm, 3JH,H=8.9 Hz, 2H, 7-CH), 4.80 (dm, 2JH,F=47.3 Hz, 2H, 11-CH2), 4.65 (d, 2JH,H=16.0 Hz, 1H, 12-CHA), 4.56 (t, 3JH,H=7.3 Hz, 1H, 2-CH), 4.53 (dd, 3JH,F=17.1 Hz, 2JH,H=3.0 Hz, 1H, 5-CHcis), 4.29 (d, 2JH,H=15.7 Hz, 2H, 12-CHB), 4.25 (dm, 3JH,F=27.8 Hz, 2H, 10-CH2), 4.11 (dd, 3JH,F=, 49.9 Hz, 2JH,H=3.0 Hz, 1H, 5-CHtrans), 2.70 (dt, 3JH,F=15.3 Hz, 3JH,H=6.9 Hz, 1H, 3-CHA), 2.46 (ddd, 3JH,F=20.0 Hz, 2JH,H=15.1 Hz, 3JH,H=7.6 Hz, 1H, 3-CHB), 1.36 (s, 9H, 18-CH3). 13C NMR (75 MHz, CDCl3): δ=168.5 (s, C-1), 161.9 (ds, 1JH,F=256.5 Hz, C-4), 161.7 (s, C-9), 136.7 (s, C-13), 132.5 (s, C-6), 129.9 (d, C-8), 128.5 (d, C-14 o. C-15), 128.4 (d, C-14 o. C-15), 127.7 (d, C-16), 114.6 (d, C-7), 93.1 (dt, 2JH,F=19.2 Hz, C-5), 82.6 (s, C-17), 81.5 (dt, 1JC,F=171.6 Hz, C-11), 67.4 (dt, 2JC,F=20.2 Hz, C-10), 57.6 (d, C-2), 50.0 (t, C-12), 34.1 (dt, 2JC,F=27.9 Hz, C-3), 27.8 (q, C-18). 19F NMR (282 MHz, CD3CN): δ =−97.7 (m, 3JH,F=49.8 Hz, 3JH,F=19.8 Hz, 3JH,F=16.4.0 Hz, 4-CF), −224.3 (tt, 2JH,F=47.3 Hz, 3JH,F=27.6 Hz, 11-CH2F). Exact mass (ESI+): C24H29F2NO5S+Na+, calcd. 504.1632. found 504.1613. (C24H29F2NO5S)2+Na+, calcd. 985.3367. found 985.3345. Optical rotation: [α]58920=+19.9, [α]57820=+20.3, [α]54620=+23.2, [α]43620=+42.5, [α]36520=+74.0 (c=1.018, CHCl3).
Prepared from tert-butyl N-[p-(2-chloroethoxy)phenylsulfonyl]-2-amino-4-fluoropent-4-enoate (816 mg 2.0 mmol), benzyl bromide (342 mg, 2.0 mmol) and potassium carbonate 2.8 g (20.0 mmol). Yield: 580 mg (58%). 1H NMR (300 MHz, CDCl3): δ=7.82 (dm, 3JH,H=8.9 Hz, 2H, 8-CH), 7.29 (m, 5H, 14-CH, 15-CH, 16-CH), 6.96 (dm, 3JH,H=9.0 Hz, 2H, 7-CH), 4.65 (d, 2JH,H=15.8 Hz, 1H, 12-CHA), 4.56 (t, 3JH,H=7.3 Hz, 1H, 2-CH), 4.53 (dd, 3JH,F=17.2 Hz, 2JH,H=2.9 Hz, 1H, 5-Ccis), 4.29 (d, 2JH,H=15.7 Hz, 1H, 12-CHB), 4.27 (t, 3JH,H=5.6 Hz, 1H, 10-CH2), 4.11 (dd, 3JH,F=49.9 Hz, 2JH,H=3.0 Hz, 1H, 5-CHtrans), 3.84 (t, 3JH,H=5.7 Hz, 2H, 11-CH2Cl), 2.70 (td, 2JH,H=15.4 Hz, 3JH,H=7.0 Hz, 1H, 3-CHA), 2.46 (ddd, 3JH,F=20.0 Hz, 2JH,H=15.2 Hz, 3JH,H=7.6 Hz, 1H, 3-CHB), 1.36 (s, 9H, 18-CH3). 13C NMR (75 MHz, CDCl3): δ=168.5 (s, C-1), 161.9 (ds, 1JH,F=256.5 Hz, C-4), 161.5 (s, C-9), 136.7 (s, C-13), 132.6 (s, C-6), 129.9 (d, C-8), 128.5 (d, C-14 o. C-15), 128.4 (d, C-14 o. C-15), 127.7 (d, C-16), 114.6 (d, C-7), 93.1 (dt, 2JC,F=18.9 Hz, C-5), 82.6 (s, C-17), 68.2 (t, C-10), 57.6 (d, C-2), 49.9 (t, C-12), 41.5 (t, C-11), 34.1 (dt, 2JH,F=28.2 Hz, C-3), 27.8 (q, C-18). 19F NMR (282 MHz, CDCl3): δ =−97.7 (ddt, 3JH,F=49.9 Hz, 3JH,F=19.9 Hz, 3JH,F=16.4 Hz, 4-CF). Exact mass (ESI+): C24H29ClFNO5S+Na+, calcd. 520.1337. found 520.1314; (C24H29ClFNO5S)2+Na+, calcd. 1017.2776. found 1017.2708. Optical rotation: [α]58920=−24.8, [α]57820=−26.2, [α]54620=−30.0, [α]43620=−54.4, [α]36520=−95.0 (c=1.038, CHCl3).
Prepared from tert-butyl N-[p-(2-chloroethoxy)phenylsulfonyl]-2-amino-4-fluoropent-4-enoate (1.50 g, 3.68 mmol), benzyl bromide (630 mg, 3.68 mmol) and potassium carbonate (5.0 g, 36.8 mmol). Yield: 1.28 g (70%). 1H NMR (300 MHz, CDCl3): δ=7.82 (dm, 3JH,H=9.0 Hz, 2H, 8-CH), 7.28 (m, 5H, 14-CH, 15-CH, 16-CH), 6.96 (dm, 3JH,H=9.0 Hz, 2H, 7-CH), 4.66 (d, J=15.8, 1H, 12-CHA), 4.56 (m, 1H, 2-CH), 4.53 (dd, J=17.1 Hz, 3.0 Hz, 2H, 5-CHcis), 4.29 (d, J=15.7 Hz, 1H, 12-CHB), 4.28 (t, J=5.6 Hz, 2H, 10-CH2), 4.11 (dd, J=49.9 Hz, 3.0 Hz, 1H, 5-CHtrans), 3.83 (t, J=5.8 Hz, 2H, 11-CH2Cl), 2.70 (td, J=15.4 Hz, 6.9 Hz, 1H, trans, 3-CHA), 2.46 (ddd, J=20.1 Hz, 15.2 Hz, 7.6 Hz, 1H, 3-CHB), 1.36 (s, 9H, 18-CH3). 13C NMR (75 MHz, CDCl3): δ=168.6 (s, C-1), 161.9 (ds, 1JH,F=256.3 Hz, C-4), 161.5 (s, C-9), 136.7 (s, C-13), 132.7 (s, C-6), 129.9 (d, C-8), 128.5 (d, C-14 o. C-15), 128.4 (d, C-14 o. C-15), 127.8 (d, C-16), 114.6 (d, C-7), 93.1 (dt, 2JH,F=19.1 Hz, C-5), 82.6 (s, C-17), 68.2 (t, C-10), 57.6 (d, C-2), 50.0 (t, C-12), 41.5 (t, C-11), 34.1 (dt, 2JH,F=27.9 Hz, C-3), 27.8 (q, C-18). 19F NMR (282 MHz, CDCl3): δ =−97.7 (m, 3JH,F=49.9 Hz, 3JH,F=19.9 Hz, 3JH,F=16.7 Hz, 4-CF). Exact mass (ESI+): C24H29ClFNO5S+Na+, calcd. 520.1337. found 520.1326; (C24H29ClFNO5S)2+Na+, calcd. 1017.2776. found 1017.2748. Optical rotation: [α]58920=+21.1, [α]57820=+22.2, [α]54620=+25.1, [α]43620=+45.5, [α]36520=n.d. (c=1.005, CHCl3).
Hydrolysis of the Tert-Butylesters
In a dried YOUNG-tube the amino acid tert-butylester is dissolved in dry dichloromethane (20 mL/mmol) under argon and treated with trifluoroacetic acid (20 mL/mmol). The YOUNG-tube is flushed with argon and sealed. The mixture is stirred at r.t. for 3-4 h. Subsequently the reaction mixture is evaporated to dryness in vacuum. The residue is dissolved in chloroform (100 mL/mmol) and washed with an aqueous solution of citric acid and sodium bicarbonate (25 mL/mmol, pH≈4). The aqueous phase is extracted with chloroform (4×30 mL/mmol) and the combined organic phases are dried with magnesium sulfate. After evaporation of the solvent the crude products are obtained as colorless, viscos oils, which in high vacuum solidified. These products were used for the next step without purification.
Prepared from tert-butyl N-benzyl-N-[p-(2-fluoroethoxy)phenylsulfonyl]-2-amino-4-fluoropent-4-enoate (300 mg 0.7 mmol). 1H NMR (300 MHz, CD3CN): δ=7.78 (dm, 3JH,H=9.0 Hz, 2H, 8-CH), 7.30 (m, 5H, 14-CH, 15-CH, 16-CH), 7.04 (dm, 3JH,H=9.0 Hz, 2H, 7-CH), 4.76 (dm, 2JH,F=47.7 Hz, 2H, 11-CH2), 4.65 (dd, 3JH,H=8.5 Hz, 3JH,H=6.0 Hz, 1H, 2-CH), 4.57 (d, 2JH,H=16.1 Hz, 1H, 12-CHA), 4.52 (dd, 3JH,F=17.8 Hz, 2JH,H=3.0 Hz, 1H, 5-CHcis), 4.33 (d, 2JH,H=16.1 Hz, 1H, 12-CHB), 4.29 (dm, 3JH,F=29.5 Hz, 2H, 10-CH2), 4.14, (dd, 3JH,F=51.5 Hz, 3JH,H=3.5 Hz, 1H, 5-CHtrans 2.76 (ddd, 3JH,F=14.8 Hz, 2JH,H=13.7 Hz, 3JH,H=6.0 Hz, 1H, 3-CHA), 2.48 (ddd, 3JH,F=22.3 Hz, 2JH,H=15.4 Hz, 3JH,H=8.5 Hz, 1H, 3-CHB). 13C NMR (75 MHz, CD3CN): δ=171.2 (s, C-1), 163.2 (ds, 1JH,F=254.7 Hz, C-4), 163.1 (s, C-9), 138.2 (s, C-13), 133.0 (s, C-6), 131.0 (d, C-8), 129.6 (d, C-14 o. C-15), 129.3 (d, C-14 o. C-15), 128.7 (d, C-16), 115.7 (d, C-7), 93.8 (dt, 2JH,F=18.9 Hz, C-5), 83.1 (dt, 1JH,F=167.3 Hz, C-11), 68.8 (dt, 2JC,F=19.4 Hz, C-10), 57.9 (d, C-2), 50.8 (t, C-12), 34.1 (dt, 2JH,F=28.2 Hz, C-3). 19F NMR (282 MHz, CD3CN): δ =−97.7 (dddd, 3JH,F=51.0 Hz, 3JH,F=22.1 Hz, 3JH,F=17.7 Hz, 3JH,F=13.4 Hz, 4-CF), −224.0 (tt, 2JH,F=47.7 Hz, 3JH,F=29.5 Hz, 11-CH2F). Exact mass (ESI+): C20H31F2NO5S+Na+, calcd. 448.1006. found 448.1004, (C20H31F2NO5S)2+Na+, calcd. 873.2115. found 873.2116. Exact mass (ESI−): C20H31F2NO5S—H+, calcd. 424.1036. found 424.1057, (C20H31F2NO5S)2—H+, calcd. 849.2144. found 849.2152.
Prepared from tert-butyl N-benzyl-N-[p-(2-fluoroethoxy)phenylsulfonyl]-2-amino-4-fluoropent-4-enoate (400 mg, 0.94 mmol). The product is contaminated with some starting material. 1H NMR (300 MHz, CD3CN): δ=7.78 (dm, 3JH,H=9.1, 2H, 8-CH), 7.29 (m, 5H, 14-CH, 15-CH, 16-CH), 7.04 (dm, 3JH,H=9.0, 2H, 7-CH), 4.75 (dm, 2JH,F=47.7 Hz, 2H, 11-CH2), 4.65 (dd, 3JH,H=8.5, 3JH,H=6.0, 1H, 2-CH), 4.57 (d, 2JH,H=16.1, 1H, 12-CHA), 4.52 (dd, 3JH,F=17.6, 2JH,H=3.0, 1H, 5-CHcis), 4.32 (d, 2JH,H=16.1, 1H, 12-CHB), 4.30 (dm, JH,F=29.4 Hz, 2H, 10-CH2), 4.14 (dd, 3JH,F=51.8, 2JH,H=3.4, 1H, 5-CHtrans, 2.76 (ddd, 3JH,F=14.7, 2JH,H=13.7, 3JH,H=6.0, 1H, 3-CHA), 2.47 (ddd, 3JH,F=22.2, 2JH,H=15.4, 3JH,H=8.5, 1H, 3-CHB). 13C NMR (75 MHz, CD3CN): δ=171.2 (s, C-1), 163.2 (ds, 1JH,F=254.7 Hz, C-4), 163.1 (s, C-9), 138.2 (s, C-13), 133.0 (s, C-6), 131.0 (d, C-8), 129.6 (d, C-14 o. C-15), 129.3 (d, C-14 o. C-15), 128.7 (d, C-16), 115.7 (d, C-7), 93.8 (dt, 2JH,F=18.9 Hz, C-5), 83.1 (dt, 1JH,F=167.5 Hz, C-11), 68.8 (dt, 2JC,F=19.2 Hz, C-10), 57.9 (d, C-2), 50.8 (t, C-12), 34.2 (dt, 2JH,F=28.0 Hz, C-3). 19F NMR (282 MHz, CD3CN): δ =−97.7 (dddd, 3JH,F=50.9 Hz, 3JH,F=22.1 Hz, 3JH,F=17.7 Hz, 3JH,F=13.3 Hz, 4-CF), −224.0 (tt, 2JH,F=47.7 Hz, 3JH,F=29.5 Hz, 11-CH2F). Exact mass (ESI+): C20H31F2NO5S+Na+, calcd. 448.1006. found 448.1009; Exact mass (ESI−): C20H31F2NO5S—H+, calcd. 424.1036. found 424.1057, (C2H31F2NO5S)2—H+, calcd. 849.2144. found 849.2157.
Prepared from tert-butyl N-benzyl-N-[p-(2-chloroethoxy)phenylsulfonyl]-2-amino-4-fluoropent-4-enoate (300 mg, 0.68 mmol). 1H NMR (300 MHz, CD3CN): δ=7.78 (d, 3JH,H=9.1 Hz, 2H, 8-CH), 7.29 (m, 5H, 14-CH, 15-CH, 16-CH), 7.03 (d, 3JH,H=9.1 Hz, 1H, 7-CH), 4.65 (dd, 3JH,H=8.5 Hz, 3JH,H=6.0 Hz, 1H, 2-CH), 4.57 (d, 2JH,H=16.1 Hz, 1H, 12-CHA), 4.52 (dd, 3JH,F=17.7 Hz, 2JH,H=3.2 Hz, 1H, 5-CHcis), 4.33 (m, 2H, 10-CH2), 4.32 (d, 2JH,H=16.1 Hz, 1H, 12-CHB), 4.14 (dd, 3JH,F=50.9 Hz, 2JH,H=3.1 Hz, 1H, 5-CHtrans), 3.90 (m, 2H, 11-CH2Cl), 2.76 (ddd, 3JH,F=16.1 Hz, 2JH,H=14.2 Hz, 3JH,H=5.9 Hz, 2H), 2.47 (ddd, 3JH,F=22.3 Hz, 2JH,H=15.5 Hz, 3JH,H=8.6 Hz, 1H). 13C NMR (75 MHz, CD3CN): δ=171.2 (s, C-1), 163.2 (d, 1JH,F=254.5 Hz, C-4), 162.9 (s, C-9), 138.2 (s, C-13), 133.1 (s, C-6), 131.0 (s, C-8), 129.6 (s, C-14 or C-15), 129.3 (s, C-14 or C-15), 128.7 (s, C-16), 115.7 (s, C-7), 93.8 (d, 2JH,F=18.9 Hz, C-5), 69.6 (s, C-10), 57.9 (s, C-2), 50.8 (s, C-12), 43.5 (s, C-11), 34.1 (d, 2JC,F=28.4 Hz, C-3). 19F NMR (282 MHz, CD3CN): δ =−97.7 (dddd, 3JH,F=51.0 Hz, 3JH,F=22.2 Hz, 3JH,F=17.7 Hz, 3JH,F=13.3 Hz, 4-CF).
Prepared from tert-butyl N-benzyl-N-[p-(2-chloroethoxy)phenylsulfonyl]-2-amino-4-fluoropent-4-enoate (430 mg, 0.97 mmol). The crude product was directly used for the next step.
Synthesis of the tert-butyl hydroxamates
The N,N-disubstituted α-amino acid (1 equivalent) is suspended in dichloromethane (15 mL/mmol). 1-Hydroxy-benzotriazol (HOBT, 1 equivalent) is added and the mixture is stirred for a short period of time. Then N-methylmorpholin (NMM, 5 equivalents) is added under stirring while the carboxylic acid dissolves. Subsequently, O-tert-butylhydroxylamine (3 aquivalents) is added and the solution is stirred for 5 min. After addition of 1-(3-dimethylaminopropyl-3-ethylcarbodiimid-hydrochloride (EDC) (1.3 equivalents) the mixture is stirred at r.t. over night. The solution is washed with water (20 mL), and the aqueous phase is extracted with dichloromethane (3×20 mL). The combined organic phase is washed with brine (1×20 mL) and dried with magnesium sulfate. The solvent is evaporated and the crude product is purified by column chromatography (silica gel, Cy/EtOAc, 1:3). The silica gel has to be inactivated using the eluent mixture with 2% of triethylamine. The products are obtained as highly viscose liquids, which on drying in high vacuum give amorphous solids.
wherein R3 is CH2CH3, CH2CH2F, CH2CH═CH2, CH2CF═CH2.
Prepared from N-benzyl-N-[p-(2-fluoroethoxy)phenylsulfonyl]-2-amino-4-fluoropent-4-ene carboxylic acid (191 mg, 0.45 mmol), HOPT (62 mg, 0.45 mmol), NMM (240 mg, 2.25 mmol), O-tert-butylhydroxylamine (175 mg, 1.35 mmol) and EDC (115 mg, 0.58 mmol). Yield: 147 mg (66%). 1H NMR (300 MHz, CDCl3): δ=8.54 (s, 1H, 17-NH), 7.69 (dm, 3JH,H=9.0 Hz, 2H, 8-CH), 7.29 (m, 5H, 14-CH, 15-CH, 16-CH), 6.95 (dm, 3JH,H=9.0 Hz, 2H, 7-CH), 4.77 (dm, 2JH,F=47.3 Hz, 2H, 11-CH2), 4.64 (d, 2JH,H=15.7 Hz, 1H, 12-CHA), 4.54 (t, 3JH,H=7.2 Hz, 1H, 2-CH), 4.42 (dd, 3JH,F=17.1 Hz, 2JH,H=2.7 Hz, 2H, 5-CHcis), 4.42 (d, overlapped with dd at 4.42), 2JH,H=15.8 Hz, 1H, 12-CHB) 4.25 (m, 3JH,F=27.8 Hz, 2H, 10-CH2), 4.05 (dd, 3JH,F=50.0 Hz, 2JH,H=3.0 Hz, 1H, 5-CHtrans), 2.81 (ddd, 3JH,F=18.3 Hz, trans, 9 2JH,H=14.9 Hz, 3JH,H=7.6 Hz, 1H, 3-CHA), 2.32 (ddd, 3JH,F=19.1 Hz, 2JH,H=14.9 Hz, 3JH,H=7.0 Hz, 1H, 3-CHB), 1.22 (s, 9H, 19-CH3). 13C NMR (75 MHz, CDCl3): δ=167.0 (s, C-1), 161.9 (s, C-9), 161.3 (ds, 1JH,F=256.0 Hz, C-4), 136.5 (s, C-13), 131.7 (s, C-6), 129.6 (d, C-8), 128.7 (d, C-14 o. C-15), 128.5 (d, C-14 o. C-15), 127.8 (d, C-16), 114.7 (d, C-7), 93.7 (dt, 2JH,F=19.1 Hz, C-5), 82.3 (s, C-18), 81.4 (dt, 1JH,F=171.5 Hz, C-11), 67.3 (dt, JC,F=20.4 Hz, C-10), 55.0 (d, C-2), 48.6 (t, C-12), 32.4 (dt, 2JH,F=27.4 Hz, C-3), 26.1 (q, C-19). 19F NMR (282 MHz, CDCl3): δ =−97.92 (m, 3JH,F=50.1 Hz, 3JH,F=18.4 Hz, 4-CF), −223.79 (tt, 3JH,F=47.3 Hz, 3JH,F=27.8 Hz, 11-CH2F). Elemental analysis: C24H30F2N2O5S (M=496.57 g/mol), calcd. C, 58.05; H, 6.09; N, 5.64. found C, 57.94, H, 6.24, N, 5.57%. Exact mass (ESI+): C24H30F2N2O5S+H+, calcd. 497.1922. found 497.1908; C24H30F2N2O5S+Na+, calcd. 519.1741. found 519.1733; (C24H30F2N2O5S)2+Na+, calcd. 1015.3585. found 1015.3566. Optical rotation: [α]58920=−23.9, [α]57820=−25.1, [α]54620=−28.9, [α]43620=−52.6, [α]36520=−92.9 (c=0.993, CHCl3).
Prepared from N-benzyl-N-[p-(2-fluorethoxy)phenylsulfonyl]-2-amino-4-fluoropent-4-enoic acid (206 mg, 0.483 mmol), HOPT (65 mg, 0.483 mmol), NMM (245 mg, 2.415 mmol), O-tert-butylhydroxylamine (182 mg, 1.450 mmol) and EDC (121 mg, 0.628 mmol). Yield: 200 mg (83%). 1H NMR (300 MHz, CDCl3): δ=8.46 (s, 1H, 17-NH), 7.69 (dm, 3JH,H=8.9 Hz, 2H, 8-CH), 7.29 (m, 5H, 14-CH, 15-CH, 16-CH), 6.96 (dm, 3JH,H=9.0 Hz, 2H, 7-CH), 4.78 (dm, 2JH,F=47.4 Hz, 2H, 11-CH2), 4.64 (d, 2JH,H=15.7 Hz, 1H, 12-CHA), 4.54 (t, 3JH,H=7.2 Hz, 1H, 2-CH), 4.42 (dd, 3JH,F=17.1 Hz, 2JH,H=3.0 Hz, 2H, 5-CHcis), 4.40 (d, 2JH,H=15.6 Hz, 1H, 12-CHB) 4.27 (m, 3JH,F=27.6 Hz, 2H, 10-CH2), 4.05 (dd, 3JH,F=50.0 Hz, 2JH,H=3.0 Hz, 1H, 5-CHtrans), 2.821 (ddd, 3JH,F=18.5 Hz, 2JH,H=14.9 Hz, 3JH,H=7.6 Hz, 1H, 3-CHA), 2.31 (ddd, 3JH,F=19.1 Hz, 2JH,H=14.9 Hz, 3JH,H=6.9 Hz, 1H, 3-CHB), 1.23 (s, 9H, 19-CH3). 13C NMR (75 MHz, CDCl3): δ=167.0 (s, C-1), 161.9 (s, C-9), 161.3 (ds, 1JH,F=255.8 Hz, C-4), 136.5 (s, C-13), 131.9 (s, C-6), 129.6 (d, C-8), 128.8 (d, C-14 o. C-15), 128.6 (d, C-14 o. C-15), 128.0 (d, C-16), 114.8 (d, C-7), 93.7 (dt, 2JH,F=19.2 Hz, C-5), 82.4 (s, C-18), 81.4 (dt, 1JH,F=171.8 Hz, C-11), 67.4 (dt, 2JH,F=20.5 Hz, C-10), 55.1 (d, C-2), 48.7 (t, C-12), 32.4 (dt, 2JC,F=27.6 Hz, C-3), 26.2 (q, C-19). 19F NMR (282 MHz, CDCl3): δ =−98.51 (m, 3JH,F=50.3 Hz, 3JH,F=18.0 Hz, 4-CF), −224.35 (tt, 2JH,F=47.3 Hz, 3JH,F=27.8 Hz, 11-CH2F). Elemental analysis: C24H30F2N2O5S (M=496.57 g/mol), calcd. C, 58.05; H, 6.09; N, 5.64. found C, 57.78; H, 6.24; N, 5.56%. Exact mass (ESI+): C24H30F2N2O5S+FE: calcd. 497.1922. found 497.1917; C24H30F2N2O5S+Na+, calcd. 519.1741. found 519.1733; (C24H30F2N2O5S)2+Na+, calcd. 1015.3585. found 1015.3564. Optical rotation: [α]58920=+21.7 (c=1.039, CHCl3).
Prepared from N-benzyl-N-[p-(2-chloroethoxy)phenylsulfonyl]-2-amino-4-fluoropent-4-enoic acid (243 mg, 0.55 mmol) HOBT (75 mg, 0.55 mmol), NMM (283 mg, 2.80 mmol) O-tert-butylhydroxylamine (213 mg, 1.70 mmol) and EDC (138 mg, 0.72 mmol). Yield: 204 mg (72%). 1H NMR (300 MHz, CDCl3): δ=8.48 (s, 1H, 17-NH), 7.69 (dm, 3JH,H=8.9 Hz, 2H, 8-CH), 7.28 (m, 5H, 14-CH, 15-CH, 16-CH), 6.94 (dm, 3JH,H=8.8 Hz, 2H, 7-CH), 4.64 (d, 2JH,H=15.7 Hz, 1H, 12-CHA), 4.53 (t, 3JH,H=7.2 Hz, 1H, 2-CH), 4.42 (dd, 3JH,F=17.1 Hz, 2JH,H=3.0 Hz, 1H, 5-CHcis), 4.41 (d, 2JH,H=15.5 Hz, 1H, 12-CHB), 4.28 (t, 3JH,H=5.7 Hz, 2H, 10-CH2), 4.06 (dd, 3JH,F=49.9 Hz, 2JH,H=3.1 Hz, 1H, 5-CHtrans), 3.84 (t, J=5.7 Hz, 2H, 11-CH2), 2.82 (ddd, 3JH,F=18.5 Hz, 2JH,H=14.9 Hz, 3JH,H=7.6 Hz, 1H, 3-CHA), 2.31 (ddd, 3JH,F=18.9 Hz, 2JH,H=14.9 Hz, 3JH,H=6.9 Hz, 1H, 3-CHB), 1.23 (s, 9H, 19-CH3). 13C NMR (75 MHz, CDCl3): δ=167.0 (s, C-1), 161.8 (s, C-9), 161.3 (ds, 1JH,F=255.8, C-4), 136.4 (s, C-13), 131.9 (s, C-6), 129.7 (d, C-8), 128.8 (d, C-14 o. C-15), 128.6 (d, C-14 o. C-15), 127.9 (d, C-16), 114.8 (d, C-7), 93.7 (dt, 2JC,F=19.4, C-5), 82.4 (s, C-18), 68.2 (t, C-10), 55.0 (d, C-2), 48.7 (t, C-12), 41.4 (t, C-11), 32.4 (dt, 2JH,F=27.6 Hz, C-3), 26.1 (q, C-19). 19F NMR (282 MHz, CDCl3): δ =−98.49 (m, 3JH,F=49.9 Hz, 3JH,F=18.9 Hz, 3JH,F=17.2 Hz 4-CF). Elemental analysis: C24H3ClFN2O5S (M=513.02 g/mol), calcd. C, 56.19; H, 5.89; N, 5.46. found C, 55.55; H, 5.89; N, 5.34%. Exact mass (ESI+): C24H30ClFN2O5S+Na+, calcd. 535.1446. found 535.1434. Optical rotation: [α]58920=−2.2, [α]36520=−6.9 (c=1.025 CHCl3).
Prepared from N-benzyl-N-[p-(2-chloroethoxy)phenylsulfonyl]-2-amino-4-fluoropent-4-enoic acid (348 mg, 0.79 mmol), HOBT (107 mg, 0.79 mmol), NMM (405 mg, 3.94 mmol), O-tert-butylhydroxylamine (305 mg, 2.36 mmol) and EDC (198 mg, 1.20 mmol). Yield: 302 mg (75%). 1H NMR (300 MHz, CDCl3): δ=8.48 (s, 1H, 17-NH), 7.69 (dm, 3JH,H=8.9 Hz, 2H, 8-CH), 7.28 (m, 5H, 14-CH, 15-CH, 16-CH), 6.94 (dm, 3JH,H=8.9 Hz, 2H, 7-CH), 4.64 (d, 2JH,H=15.6 Hz, 1H, 12-CHA), 4.53 (t, 3JH,H=7.2 Hz, 1H, 2-CH), 4.43 (dd, 3JH,F=17.1 Hz, 2JH,H=3.0 Hz, 1H, 5-CHcis), 4.40 (d, 2JH,H=15.6 Hz, 1H, 12-CHB), 4.28 (t, 3JH,H=5.7 Hz, 2H, 10-CH2), 4.06 (dd, 3JH,F=49.6 Hz, 2JH,H=3.5 Hz, 1H, 5-CHtrans), 3.84 (t, J=5.7 Hz, 2H, 11-CH2), 2.82 (ddd, 3JH,F=18.6 Hz, 2JH,H=14.9 Hz, 3JH,H=7.6 Hz, 1H, 3-CHA), 2.31 (ddd, 3JH,F=18.9 Hz, 2JH,H=14.9 Hz, 3JH,H=6.8 Hz, 1H, 3-CHB), 1.23 (s, 9H, 19-CH3). 13C NMR (75 MHz, CDCl3): δ=167.0 (s, C-1), 161.8 (s, C-9), 161.3 (ds, 1JH,F=256.1, C-4), 136.4 (s, C-13), 131.9 (s, C-6), 129.7 (d, C-8), 128.8 (d, C-14 o. C-15), 128.6 (d, C-14 o. C-15), 127.9 (d, C-16), 114.8 (d, C-7), 93.7 (dt, 2JC,F=19.2, C-5), 82.4 (s, C-18), 68.2 (t, C-10), 55.0 (d, C-2), 48.7 (t, C-12), 41.4 (t, C-11), 32.4 (dt, 2JH,F=27.1 Hz, C-3), 26.1 (q, C-19). 19F NMR (282 MHz, CDCl3): δ =−98.50 (m, 3JH,F=49.9 Hz, 3JH,F=18.5 Hz 4-CF). Elemental analysis: C24H30ClFN2O5S (M=513.02 g/mol), calcd. C, 56.19; H, 5.89; N, 5.46. found C, 55.20, H, 5.88, N, 5.36%. Exact mass (ESI+): C24H30ClFN2O58 +Na+, calcd. 535.1446. found 535.1434. Optical rotation: [α]58920=+21.5, [α]57820=+22.2, [α]54620=+25.5, [α]43620=+46.5, [α]36520=+82.2 (c=0.999, CHCl3).
General Procedure for the Preparation of Tetrahydropyranyl (THP)-Protected Hydroxamic Acids
To a solution of carboxylic acid (100 mg, 0.235 mmol) in DMF (0.06 mmol/mL, 1 mL) 1-hydroxybenzotriazole hydrate (HOBT, 1.2 eq., 38 mg, 0.282 mmol), 4-methylmorpholine (NMM, 3.0 eq., 78 μL, 249.70 mmol), O-tetrahydro-2H-pyran-2-yl-hydroxylamine (3.1 eq., 85 mg, 0.729 mmol and N-[dimethylamino)-propyl]-N′-ethylcarbodiimide hydrochloride (EDC, 1.4 eq., 63 mg, 0.329 mmol) were added. After stirring overnight at room temperature the reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (3×5 mL). The combined organic phases were washed successively with water, 5% aqueous KHSO4, saturated aqueous NaHCO3 and brine, and dried over magnesium sulfate. After removing the solvent under reduced pressure column chromatographic purification (silica gel, cyclohexane/ethyl acetate 2:1) yielded the THP-protected hydroxamic acid.
Light brown wax; yield: 92 mg (75%). 1H NMR (300 MHz, CDCl3); δ 9.19 (s, NH, 1H), 9.15 (s, NH, 1H), 7.77 (d, 3JH,H=8.9 Hz, PhH, 2H), 7.72 (d, 3JH,H=9.0 Hz, PhH, 2H), 7.39-7.21 (m, PhH, 5H), 6.98 (d, 3JH,H=9.0 Hz, PhH, 2H), 6.97 (d, 3JH,H=9.0 Hz, PhH, 2H), 4.92-4.88 (m, NHOCHO), 4.79 (dm, 2JH,F=47.4 Hz, CH2F, 2H), 4.66 (AB, d, 2JH,H=16.0 Hz, NCH2, 1H), 4.58-4.45 (m, NCH, NCH2, 2H), 4.43 (dd, CFCH2, Hcis, 3JH,F=17.1 Hz, 2JH,F=3.0 Hz, 1H), 4.39 (dd, CFCH2, Hcis, 3JH,F=17.1 Hz, 2JH,F=3.0 Hz, 1H), 4.27 (dm, 3JH,F=27.7 Hz, O—CH2CH2F, 2H), 4.10 (dd, CFCH2, Htrans, 3JH,F=49.9 Hz, 2JH,F=3.1 Hz, 1H), 3.95-3.84 (m, NHOCHOCH2, 1H), 3.70-3.55 (m, NHOCHOCH2, 1H), 2.93-2.58 (AB, m, NCHCH2, 1H), 2.47-2.18 (AB, m, NCHCH2, 1H), 1.88-1.50 (m, THP-CH2, 6H). 13C NMR (75 MHz, CDCl3): δ 165.8 (CONH), 165.6 (CONH), 162.0 (q, PhCOCH2CH2F), 161.2 (d, 1JH,F=256.3 Hz, CFCH2), 161.2 (d, 1JH,F=256.0 Hz, CFCH2), 136.6 (qPhCCH2N), 136.4 (qPhCCH2N), 131.5 (qPhCSO2), 129.7 (PhCH), 129.7 (PhCH), 128.7 (PhCH), 128.6 (PhCH), 128.5 (PhCH), 128.4 (PhCH), 127.8 (PhCH), 127.8 (PhCH), 114.7 (PhCH), 114.7 (PhCH), 102.1 (NHOCHO), 101.7 (NHOCHO), 93.7 (d, 2JH,F=19.0 Hz, CFCH2), 93.6 (d, 2JH,F=18.9 Hz, CFCH2), 81.4 (1JH,F=171.6 Hz, OCH2CH2F), 67.4 (d, 2JH,F=20.5 Hz, OCH2CH2F), 62.0 (NHOCHOCH2), 54.7 (NCH), 54.5 (NCH), 48.6 (NCH2), 48.5 (NCH2), 32.3 (d, 2JC,F=27.6 Hz, NCHCH2CF), 32.0 (d, 2JH,F=26.9 Hz, NCHCH2CF), 27.7 (NHOCHCH2), 27.7 (NHOCHCH2), 24.9 (CH2), 18.2 (NHOCHCH2CH2). MS-ES-EM: m/z=547.1683 [M+Na)+] calcd for C25H30F2N2O6SNa+: 547.1685.
Light brown wax; yield: 101 mg (81%). 1H NMR (300 MHz, CDCl3): δ 9.08 (s, NH, 1H), 7.77 (d, 3JH,H=9.0 Hz, PhH, 2H), 7.72 (d, 3JH,H=9.0 Hz, PhH, 2H), 7.39-7.21 (m, PhH, 5H), 6.98 (d, 3JH,H=9.0 Hz, PhH, 2H), 6.97 (d, 3JH,H=9.0 Hz, PhH, 2H), 4.92-4.88 (m, NHOCHO), 4.80 (dm, 2JH,F=47.3 Hz, CH2F, 2H), 4.66 (AB, m, NCH2, 1H), 4.58-4.45 (m, NCH, NCH2, 2H), 4.45-4.34 (m, CFCH2, Hcis, 1H), 4.28 (dm, 3JH,F=27.7 Hz, O—CH2CH2F, 2H), 4.10 (dd, CFCH2, Htrans, 3JH,F=49.9 Hz, 2JH,F=3.0 Hz, 1H), 3.95-3.84 (m, NHOCHOCH2, 1H), 3.70-3.58 (m, NHOCHOCH2, 1H), 2.92-2.64 (AB, m, NCHCH2, 1H), 2.43-2.18 (AB, m, NCHCH2, 1H), 1.88-1.50 (m, THP-CH2, 6H). 13C NMR (75 MHz, CDCl3): δ 165.8 (CONH), 165.7 (CONH), 162.0 (qPhCOCH2CH2F), 161.3 (d, 1JH,F=256.1 Hz, CFCH2), 136.6 (qPhCCH2N), 136.4 (qPhCCH2N), 131.6 (qPhCSO2), 129.8 (PhCH), 129.8 (PhCH), 128.8 (PhCH), 128.7 (PhCH), 128.5 (PhCH), 128.5 (PhCH), 127.9, (PhCH), 127.9 (PhCH), 114.8 (PhCH), 114.8 (PhCH), 102.2 (NHOCHO), 101.8 (NHOCHO), 93.8 (d, 2JC,F=18.9 Hz, CFCH2), 81.4 (1JC,F=171.9 Hz, OCH2CH2F), 67.4 (d, 2JH,F=20.5 Hz, OCH2CH2F), 62.1 (NHOCHOCH2), 62.1 (NHOCHOCH2), 54.7 (NCH), 54.6 (NCH), 48.7 (NCH2), 48.5 (NCH2), 32.2 (d, 2JH,F=27.6 Hz, NCHCH2CF), 27.7 (NHOCHCH2), 24.9 (CH2), 18.2 (NHOCHCH2CH2). 19F NMR (282 MHz, CDCl3): δ −97.51 and 97.77 (m, CFCH2, 1F), −223.89 and 223.90 (tt, 2JH,F=47.3, 3JH,F=27.6 Hz, OCH2CH2F, 1F). MS-ES-EM m/z=547.1695 [M+Na)+] calcd for C25H30F2N2O6SNa+: 547.1685.
General Procedure for the Acidic Hydrolysis of the THP-Protected Hydroxamic Acids
To a solution of THP-protected hydroxamic acid in 1,4-dioxane (0.5 mL/0.18 mmol) 4 N hydrochloric acid in 1,4-dioxane (4 eq.) and methanol (0.5 mL/0.18 mmol) were added and the reaction mixture was stirred at room temperature. The reaction progress was monitored by TLC (ethyl acetate). After complete conversion the reaction mixture was diluted with ethyl acetate (20 mL), washed with water (3×10 mL) and dried (MgSO4). The solvent was removed under reduced pressure and the product was purified by column chromatography (silica gel, cyclohexane/ethyl acetate 2:1).
The reaction was carried out in a 0.175 mmol scale with (2S)-2-(N-benzyl-4-(2-fluoroethoxy)phenylsulfonamido)-4-fluoro-N-[(tetrahydro-2H-pyran-2-yl)oxy]pent-4-enamide (HUG 72). After column chromatography a light brown wax was obtained. Yield: 33 mg (43%). 1H NMR (400 MHz, CDCl3): δ 9.19 (s, 1 OH or NH), 7.72 (d, 3JH,H=8.9 Hz, PhH, 2H), 7.35-7.25 (m, PhH, 5H), 6.97 (d, 3JH,H=8.9 Hz, PhH, 2H), 4.79 (dm, 2JH,F=47.3 Hz, CH2F, 2H), 4.54 (AB, d, 2JH,H=15.6 Hz, NCH2, 1H), 4.52 (m, NCH, 1H), 4.46 (AB, d, 2JH,H=15.7 Hz, NCH2, 1H), 4.38 (dd, CFCH2, Hcis, 3JH,F=17.0 Hz, 2JH,F=2.9 Hz, 1H), 4.26 (dm, 3JH,F=27.6 Hz, CH2FCH2, 2H), 4.08 (dd, CFCH2, Htrans, 3JH,F=49.7 Hz, 2JH,F=3.1 Hz, 1H), 2.85-2.69 (AB, m, NCHCH2, 1H), 2.38 (AB, m, NCHCH2, 1H). 13C NMR (101 MHz, CDCl3): δ 166.4 (CONH), 162.1 (qPhCOCH2CH2F), 161.1 (d, 1JH,F=256.3 Hz, CFCH2), 136.1 (qPhCCH2N), 131.3 (qPhCSO2), 129.7 (PhCH), 128.6 (PhCH), 128.6 (PhCH), 128.0 (PhCH), 114.8 (PhCH), 93.8 (d, 2JH,F=18.9 Hz, CFCH2), 81.4 (d, 1JC,F=171.7 Hz, OCH2CH2F), 67.4 (d, 2JH,F=20.4 Hz, OCH2CH2F), 54.2 (NCH), 48.7 (NCH2), 31.7 (d, 2JC,F=27.7 Hz, NCHCH2CF). 19F NMR (282 MHz, CDCl3): δ −97.58 (m, CFCH2, 1F), −223.84 (tt, 2JH,F=47.3, 3JH,F=27.6 Hz, OCH2CH2F, 1F). MS-ES-EM: m/z=463.11201[(M+Na)+] calcd for C20H22F2N2O5SNa+: 463.1110. HPLC tR=9.05 min (100%).
The reaction was carried out in a 0.190 mmol scale with (2R)-2-[N-benzyl-4-(2-fluoroethoxy)phenylsulfonamido]-4-fluoro-N-[(tetrahydro-2H-pyran-2-yl)oxy]pent-4-enamide (HUG 71). After column chromatography a light brown wax was obtained. Yield: 52 mg (62%). 1H NMR (400 MHz, CDCl3): δ 9.25 (s, 1OH or NH), 7.71 (d, 3JH,H=8.8 Hz, PhH, 2H), 7.38-7.18 (m, PhH, 5H), 6.96 (d, 3JH,H=8.8 Hz, PhH, 2H), 4.79 (dm, 2JH,F=47.2 Hz, CH2F, 2H), 4.54 (AB, d, 2JH,H=15.5 Hz, NCH2, 1H), 4.52 (m, NCH, 1H), 4.46 (AB, d, 2JH,H=15.8 Hz, NCH2, 1H), 4.38 (dd, CFCH2, Hcis, 3JH,F=17.0 Hz, 2JH,F=2.8 Hz, 1H), 4.26 (dm, 3JH,F=27.3 Hz, CH2FCH2, 2H), 4.08 (dd, CFCH2, H trans, 3JH,F 50.2 Hz, 2JH,F=2.2 Hz, 1H), 2.86-2.63 (AB, m, NCHCH2, 1H), 2.47-2.25 (AB, m, NCHCH2, 1H). 13C NMR (101 MHz, CDCl3): δ 166.4 (CONH), 162.1 (qPhCOCH2CH2F), 161.1 (d, 1JH,F=256.3 Hz, CFCH2), 136.2 (qPhCCH2N), 131.3 (qPhCSO2), 129.7 (PhCH), 128.6 (PhCH), 128.6 (PhCH), 128.0 (PhCH), 114.8 (PhCH), 93.7 (d, 2JH,F=19.4 Hz, CFCH2), 81.4 (d, 1JH,F=171.6 Hz, OCH2CH2F), 67.4 (d, 2JH,F=20.4 Hz, OCH2CH2F), 54.2 (NCH), 48.7 (NCH2), 31.8 (d, 2JC,F=27.8 Hz, NCHCH2CF). 19F NMR (282 MHz, CDCl3): δ −97.54 (ddd, 3JH,F=49.9 Hz, 3JH,F=36.9 Hz, 3JH,F=17.1 Hz, CFCH2, 1F), −223.83 (tt, 2JH,F=47.3, 3JH,F=27.6 Hz, OCH2CH2F, 1F). MS-ES-EM: m/z=463.11049 [(M+Na)+] calcd for C20H22F2N2O5SNa+: 463.11097. HPLC tR=9.10 min (99%).
The synthetic broad-spectrum fluorogenic substrate (7-methoxycoumarin-4-yl)acetyl pro-Leu-Gly-Leu-(3-(2,4-dinitrophenyl)-L-2,3-diamino-propionyl)-Ala-Arg-NH2 (R & D Systems) was used to assay MMP-2 and MMP-9 activity.
The inhibition of MMP-2 and MMP-9 by the MMP inhibitors of table 1 was assayed by incubating either MMP-2 (2 nM) or MMP-9 (2 nM) and the compounds to be assayed at different concentrations (10 pM-1 mM) for 30 mM at 37° C. in 50 mM Tris.HCl (pH=7.5), 0.2M NaCl, 5 mM CaCl2, 20 μM ZnSO4 and 0.05% Brij35.
An aliquot of substrate (10 μI of a 50 μM solution) was then added to 90 μM of pre-incubated MMP/compound mixture, and activity was determined at 37° C. by following product release with time. The fluorescence changes were monitored using a Fusion Universal Microplate Analyzer (Packard Bioscience) with excitation and emission wavelengths set to 330 and 390 nm, respectively. Reaction rates were measured from the initial 10 mM of the reaction profile where product release was linear with time and plotted as a function of inhibitor dose. From the resulting inhibition curves, the IC50 values for each inhibitor were calculated by nonlinear regression analysis, performed using the GRACE 5.1.8 software (Linux).
(S)-CGS-27023A
CGS-27023A
(S)-22a
(R)-22b
(S)-23a
(R)-23b
(S)-21a
(R)-21b
(S)-33a (HUG 74)
(R)-33b (HUG 78)
1)calculated from the Ki values
As can be observed from the data reported in Table 1, the compounds of the invention show an improved activity with respect to the reference compounds CGS-27023A. This effect is remarkable also in the (S)-enantiomer of the compounds of the invention.
Having thus descried in detail advantageous embodiments of the present invention, it is to be understood that the invention defined by the above paragraphs is not to be limited to particular details set forth in the above description as many apparent variations thereof are possible without departing from the spirit or scope of the present invention.
| Number | Date | Country | Kind |
|---|---|---|---|
| 11165157.6 | May 2011 | EP | regional |
