Patent Application
20090264499
This invention relates to certain 1-cyano-3-pyrrolidinyl-benzenesulfonamides that are cathepsin C inhibitors, and their use in the treatment of diseases mediated by the cathepsin C enzyme such as chronic obstructive pulmonary disease.
Cathepsins are a family of enzymes included in the papain superfamily of cysteine proteases. Cathepsins B, C, F, H, K, L, S, V, and X have been described in the scientific literature. Cathepsin C is also known in the literature as Dipeptidyl Peptidase I or “DPPI.”
A number of recently published studies have begun to describe the role cathepsin C plays in certain inflammatory processes. See E.g. Adkison et al., The Journal of Clinical Investigation 109:363-371 (2002); Tran et al., Archives of Biochemistry and Biophysics 403:160-170 (2002); Thiele et al., The Journal of Immunology 158: 5200-5210 (1997); Bidere et al., The Journal of Biological Chemistry 277: 32339-32347 (2002); Mabee et al., The Journal of Immunology 160: 5880-5885; McGuire et al., The Journal of Biological Chemistry, 268: 2458-2467; and Paris et al., FEBS Letters 369: 326-330 (1995). From these studies, it appears that cathepsin C is co-expressed with certain serine proteases, which are released from inflammatory cells recruited to cites of inflammation, and acts as a physiological activator of these proteases. Once activated, these proteases are capable of degrading various extracellular matrix components, which can lead to tissue damage and chronic inflammation.
For example, Chronic Obstructive Pulmonary Disease (“COPD”) is a chronic inflammatory disease where cathepsin C appears to play a role. The American Thoracic Society defines COPD as “a disease characterized by the presence of airflow obstruction due to chronic bronchitis or emphysema; the airflow obstruction is generally progressive, may be accompanied by airway hyperreactivity, and may be partially reversible.” American Journal of Respiratory and Critical Care Medicine 152: S77-S120 (1995). Chronic bronchitis is generally characterized by a chronic productive cough, whereas emphysema is generally characterized by permanent enlargement of the airspaces distal to the terminal bronchioles and airway wall destruction. Chronic bronchitis and emphysema usually occur together in COPD patients.
Cigarette smoking is a significant risk factor for developing COPD. Exposure to cigarette smoke and other noxious particles and gases may result in chronic inflammation of the lung. In response to such exposure, inflammatory cells such as CD8+ Tcells, macrophages, and neutrophils are recruited to the area. These recruited inflammatory cells release proteases, which are believed to play a major role in the disease etiology by degrading airway walls. Proteases believed to be involved in this process include the serine proteases neutrophil elastase (“NE”), chymase (“CY”), cathepsin G, proteinase 3 and granzymes A and B. Cathepsin C appears to be involved in activating these enzymes.
Rheumatoid arthritis (“RA”) is another chronic inflammatory disease where cathepsin C appears to play a role. Neutrophils are recruited to the site of joint inflammation and release cathepsin G, NE, and proteinase 3, which are believed to be responsible for cartilage destruction associated with RA. Cathepsin C appears to be involved in activating these enzymes.
Other conditions where cathepsin C may play a role include osteoarthritis, asthma, and Multiple Sclerosis. See E.g. Matsui K. Yuyama N. Akaiwa M. Yoshida N L. Maeda M. Sugita Y. Izuhara K., Identification of an alternative splicing variant of cathepsin C/dipeptidyl-peptidase I, Gene. 293(1-2):1-7, 2002 Jun. 26; Wolters P J. Laig-Webster M. Caughey G H., Dipeptidyl peptidase I cleaves matrix-associated proteins and is expressed mainly by mast cells in normal dog airways, American Journal of Respiratory Cell & Molecular Biology. 22(2):183-90, 2000.
One approach to treating these conditions is to inhibit the activity of the serine proteases involved in the inflammatory process, especially NE activity. See E.g., Ohbayashi, “Neutrophil elastase inhibitors as treatment for COPD”, Expert Opin. Investig. Drugs 11(7): 965-980 (2002); Shapiro, “Neutrophil Elastase: Path Clearer, Pathogen Killer, or Just Pathologic?”, Am. J. Respir. Cell Mol. Biol. 26: 266-268 (2002). In light of the role cathepsin C plays in activating certain serine proteases, especially NE, it is desirable to prepare compounds that inhibit its activity, which thereby inhibit serine protease activity. Thus, there is a need to identify compounds that inhibit cathepsin C, which can be used in the treatment of a variety of conditions mediated by cathepsin C.
In a first instance this invention relates to compounds of formula (IA) or (IB)
and a salt thereof wherein
n is 1-5,
each R1 is independently halo; OR2; C1-C10alkyl unsubstituted, or substituted by halo or a unsubstituted or substituted heteraromatic ring containing 1-3 heteroatoms selected from the group consisting of N, O or S;; CN; C(O)NR3R4; NO2; NHC(O)NR5R6; NR7R8; a heteraromatic ring containing 1-3 heteroatoms selected from the group consisting of N, O or S; aryl, unsubstituted or substituted by halo or C1-C6 alkyl; C3-C6cycloalkyl; C4-C6cycloalkenyl; C1-C10alkenyl; C1-C10alkynyl; or C(O)OR2;
R2 is H, C1-C6 alkyl, haloC1-C6 alkyl, C(O)R9;
R3 is H; C1-C6 alkyl; C3-C6cycloalkyl; or C3-C6heterocycloalkyl containing N, O or S and where when the ring heteroatom is N it is optionally substituted by CN;
R4 is H, or C1-C6 alkyl;
R5 and R6 are independently H; C1-C6alkyl; aryl unsubstituted or substituted by halo, C1-C4alkoxy, C1-C4alkyl, or OR2; C1-C10 alkyl unsubstituted or substituted by halo or C1-C4alkoxy; C3-C6cycloalkyl; C3-C6cycloalkylalkyl; C1-C10alkenyl; C1-C10alkynyl; C3-C6heteroycloalkyl; C3-C6heteroycloalkyl C1-C6alkyl; heteroarylC1-C6alkyl or heteroaryl wherein the heteroaryl ring is unsubstituted or substituted by halo, C1-C6 alkyl or halo-substituted-C1-C6 alkyl; or arylC1-C6alkyl wherein the aryl group is unsubstituted or substituted by halo, OR2, or C(O);
R7 and R8 are independently H; C1-C10alkyl; arylC1-C6alkyl wherein the aryl group is unsubstituted or substituted by R9, halo, or C1-C6alkyl; C1-C10alkyl substituted by C3-C6cycloalkyl, one or more OH groups, halo; heteroarylC1-C6alkyl; or heteroaryl; or
R9 is OH, C1-C6 alkyoxy or NR2′R3′, wherein R2′ or R3 are independently H or C1-C6alkyl.
In a second instance, this invention relates to the use of a compound of formula (IA) or (IB) or a pharmaceutically acceptable salt thereof in the prevention, management or treatment of a respiratory or inflammatory disease, such as chronic obstructive pulmonary disease or rhinitis.
In a further aspect, this invention relates to a pharmaceutically acceptable formulation comprising a compound of formula (IA) or (IB) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.
For the avoidance of doubt, unless otherwise indicated, the term “substituted” means substituted by one or more defined groups. In the case where groups may be selected from a number of alternative groups the selected groups may be the same or different.
The term “independently” means that where more than one substituent is selected from a number of possible substituents, those substituents may be the same or different.
An “effective amount” means that amount of a drug or pharmaceutical agent that will elicit the biological or medical response of a tissue, system, animal or human that is being sought, for instance, by a researcher or clinician. Furthermore, the term “therapeutically effective amount” means any amount which, as compared to a corresponding subject who has not received such amount, results in improved treatment, healing, prevention, or amelioration of a disease, disorder, or side effect, or a decrease in the rate of advancement of a disease or disorder. The term also includes within its scope amounts effective to enhance normal physiological function.
As used herein the term “alkyl” refers to a saturated straight- or branched-chain hydrocarbon radical having the specified number of carbon atoms. As used herein, the terms “C1-C6 alkyl” and “C1-C10 alkyl” refers to such a group having at least 1 and up to 6 or 10 carbon atoms respectively. Examples of such branched or straight-chained alkyl groups useful in the present invention include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, isobutyl, n-butyl, t-butyl, n-pentyl, isopentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, and n-decyl, and branched analogs of the latter 5 normal alkanes.
The term “C1-C10-alkenyl” refers to straight or branched hydrocarbon chains containing the specified number of carbon atoms and at least 1, or more, carbon-carbon double bonds. Examples include ethenyl (or ethenylene) and propenyl (or propenylene).
When the term “C1-C10alkynyl” (or “alkynylene”) is used it refers to straight or branched hydrocarbon chains containing the specified number of carbon atoms and at least 1, or more, carbon-carbon triple bonds. Examples include ethynyl (or ethynylene) and propynyl (or propynylene).
When “C3-C6cycloalkyl” is used it refers to a non-aromatic, saturated, cyclic hydrocarbon ring containing the specified number of carbon atoms. So, for example, the term “C3-C6 cycloalkyl” refers to a non-aromatic cyclic hydrocarbon ring having from three to eight carbon atoms. Exemplary “C3-C6 cycloalkyl” groups useful in the present invention include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl.
The term “C4-C6cycloalkenyl” refers to a non-aromatic monocyclic carboxycyclic ring having the specified number of carbon atoms and up to 3 carbon-carbon double bonds. “Cycloalkenyl” includes by way of example cyclopentenyl and cyclohexenyl.
“Halo” means the halogen radical fluoro, chloro, bromo, or iodo.
“Haloalkyl” means a C1-C10alkyl, C1-C10-alkenyl, or C1-C10alkynyl group that is substituted with one or more halo substituents. Haloalkyl includes trifluoromethyl.
Where “C3-C6 heterocycloalkyl” is used, it means a non-aromatic heterocyclic ring containing the specified number of ring atoms being, saturated or having one or more degrees of unsaturation and containing one or more heteroatom substitutions selected from O, S and/or N. Such a ring may be optionally fused to one or more other “heterocyclic” ring(s) or cycloalkyl ring(s). Examples of “heterocyclic” moieties include, but are not limited to, aziridine, thiirane, oxirane, azetidine, oxetane, thietane, tetrahydrofuran, pyran, 1,4-dioxane, 1,3-dioxane, piperidine, piperazine, 2,4-piperazinedione, pyrrolidine, imidazolidine, pyrazolidine, morpholine, thiomorpholine, tetrahydrothiopyran, tetrahydrothiophene, and the like.
“Aryl” refers to optionally substituted monocyclic and polycarbocyclic unfused or fused groups having 6 to 14 carbon atoms and having at least one aromatic ring that complies with Hückel's Rule. Examples of aryl groups are phenyl, biphenyl, naphthyl, anthracenyl, phenanthrenyl and the like.
“Heteroaromatic ring” means an optionally substituted aromatic monocyclic ring or polycarbocyclic fused ring system wherein at least one ring complies with Hückel's Rule, has the specified number of ring atoms, and that ring contains at least one heteratom selected from N, O, and/or S. Examples of these groups include furanyl, thiophenyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, oxo-pyridyl, thiadiazolyl, isothiazolyl, pyridinyl, pyridazinyl, pyrazinyl, pyrimidinyl, quinolinyl, isoquinolinyl, benzofuranyl, benzothiophenyl, indolyl, and indazolyl.
Compounds of particular interest include those where n in R1 is 1-4 and R1 is independently halo; OR2; C1-C10alkyl unsubstituted or substituted by halo or a unsubstituted or substituted heteraromatic ring containing 1-3 heteroatoms selected from the group consisting of N, O or S; CN; C(O)NR3R4; NO2; NHC(O)NR5R6; NR7R8; a heteraromatic ring containing 1-3 heteroatoms selected from the group consisting of N, O or S; aryl, unsubstituted or substituted by halo or C1-C6 alkyl; C3-C6cycloalkyl; C1-C10alkenyl; or C(O)OR2.
Compounds of formula (IA) of further interest are those where n in R1 is 1, or 2 or 4, and R1 is cyano, (ethylamino)carbonyl-mino, (phenylamino)carbonyl-amino, 5-methyl-1,3,4-oxadiazol-2-yl, fluorophenyl, (3-hydroxyphenyl)methyl]amino, (2,3-dihydroxypropyl)amino, (cyclopropylmethyl)amino, pentylamino, (3-methylbutyl)amino, (2-methylbutyl)amino, (3,3-dimethylbutyl)amino, (3-furanylmethyl)amino, (2-methylpentyl)amino, (3-methylpentyl)amino, nonylamino, (1,3-thiazol-2-ylmethyl)amino, (phenylmethyl)amino, (3-fluorophenyl)methylamino, (4-hydroxyphenyl)methyl-amino, 2-furanylamino, (2-chloroethyl)amino, (2,3-dihydroxypropyl)amino, butylamino, pentylamino, bis(3-methylbutyl)amino, bis(3,3-dimethylbutyl)amino, (2-methylpentyl)amino, (1,3-thiazol-2-ylmethyl)amino, (3-hydroxyphenyl)methyl, 1,2-dideoxy-D-erythro-pentitol, —NC(O)CH2-phenyl(methoxy), NC(O)phenyl(methoxy), 4-pentenamide, pentanamide, 3-methylbutanamide, 2,2-dimethylpropanamide, cyclopentanecarboxamide, 2H-pyran-4-carboxamide, 2-methylpentanamide, hexanamide, propanamide, 2-ethylbutanamide, 3-methylpentanamide, 5-methyl-2-(trifluoromethyl)-3-furancarboxamide, heptanamide, 2,2,3,3-tetrafluoropropanamide, 3-(methyloxy)propanamide, cyclobutanecarboxamide, 2-furancarboxamide, 1-methyl-1H-pyrazole-4-carboxamide, 2-methylbutanamide, 2,2-dimethylpropanamide, (2,2,2-trifluoroethyl)oxy, or C(O)OMe.
“Enantiomerically enriched” refers to products whose enantiomeric excess is greater than zero. For example, enantiomerically enriched refers to products whose enantiomeric excess is greater than 50% ee, greater than 75% ee, and greater than 90% ee.
“Enantiomeric excess” or “ee” is the excess of one enantiomer over the other expressed as a percentage. As a result, since both enantiomers are present in equal amounts in a racemic mixture, the enantiomeric excess is zero (0% ee). However, if one enantiomer was enriched such that it constitutes 95% of the product, then the enantiomeric excess would be 90% ee (the amount of the enriched enantiomer, 95%, minus the amount of the other enantiomer, 5%).
“Enantiomerically pure” means products whose enantiomeric excess is 99% ee or greater.
Some compounds of the invention have a nitrogen which is basic enough to form pharmaceutically-acceptable acid addition salts by treatment with a suitable acid. Suitable acids include pharmaceutically-acceptable inorganic acids and pharmaceutically-acceptable organic acids. Representative pharmaceutically-acceptable acid addition salts include hydrochloride, hydrobromide, nitrate, methylnitrate, sulfate, bisulfate, sulfamate, phosphate, acetate, hydroxyacetate, phenylacetate, propionate, butyrate, isobutyrate, valerate, maleate, hydroxymaleate, acrylate, fumarate, malate, tartrate, citrate, salicylate, p-aminosalicyclate, glycollate, lactate, heptanoate, phthalate, oxalate, succinate, benzoate, o-acetoxybenzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, mandelate, tannate, formate, stearate, ascorbate, palmitate, oleate, pyruvate, pamoate, malonate, laurate, glutarate, glutamate, estolate, methanesulfonate (mesylate), ethanesulfonate (esylate), 2-hydroxyethanesulfonate, benzenesulfonate (besylate), p-aminobenzenesulfonate, p-toluenesulfonate (tosylate), and napthalene-2-sulfonate.
Other iterations of compounds of the invention have an acidic functional group, one acidic enough to form salts. Representative salts include pharmaceutically-acceptable metal salts such as sodium, potassium, lithium, calcium, magnesium, aluminum, and zinc salts; carbonates and bicarbonates of a pharmaceutically-acceptable metal cation such as sodium, potassium, lithium, calcium, magnesium, aluminum, and zinc; pharmaceutically-acceptable organic primary, secondary, and tertiary amines including aliphatic amines, aromatic amines, aliphatic diamines, and hydroxy alkylamines such as methylamine, ethylamine, 2-hydroxyethylamine, diethylamine, triethylamine, ethylenediamine, ethanolamine, diethanolamine, and cyclohexylamine.
The compounds according to formula I may contain one or more asymmetric center, also referred to as a chiral center, and may, therefore, exist as individual enantiomers, diastereomers, or other stereoisomeric forms, or as mixtures thereof. Chiral centers may also be present in a substituent such as an alkyl group. Where the stereochemistry of a chiral center present in formula I, or in any chemical structure illustrated herein, is not specified the structure is intended to encompass any stereoisomer and all mixtures thereof. Thus, compounds according to formula I containing one or more chiral center may be used as racemic mixtures, enantiomerically enriched mixtures, or as enantiomerically pure individual stereoisomers.
Individual stereoisomers of a compound according to formula I which contain one or more asymmetric center may be resolved by methods known to those skilled in the art. For example, such resolution may be carried out (1) by formation of diastereoisomeric salts, complexes or other derivatives; (2) by selective reaction with a stereoisomer-specific reagent, for example by enzymatic oxidation or reduction; or (3) by gas-liquid or liquid chromatography in a chiral environment, for example, on a chiral support such as silica with a bound chiral ligand or in the presence of a chiral solvent. The skilled artisan will appreciate that where the desired stereoisomer is converted into another chemical entity by one of the separation procedures described above, a further step is required to liberate the desired form. Alternatively, specific stereoisomers may be synthesized by asymmetric synthesis using optically active reagents, substrates, catalysts or solvents, or by converting one enantiomer to the other by asymmetric transformation.
The compounds of the invention may exist in solid or liquid form. In the solid state, the compounds of the invention may exist in crystalline or noncrystalline form, or as a mixture thereof. For compounds of the invention that are in crystalline form, the skilled artisan will appreciate that pharmaceutically-acceptable solvates may be formed wherein solvent molecules are incorporated into the crystalline lattice during crystallization. Solvates may involve nonaqueous solvents such as ethanol, isopropanol, DMSO, acetic acid, ethanolamine, and ethyl acetate, or they may involve water as the solvent that is incorporated into the crystalline lattice. Solvates wherein water is the solvent that is incorporated into the crystalline lattice are typically referred to as “hydrates.” Hydrates include stoichiometric hydrates as well as compositions containing variable amounts of water. The invention includes all such solvates.
The skilled artisan will further appreciate that certain compounds of the invention that exist in crystalline form, including the various solvates thereof, may exhibit polymorphism, i.e. the capacity to occur in different crystalline structures. These different crystalline forms are typically known as polymorphs. The invention includes all such polymorphs. Polymorphs typically exhibit different melting points, IR spectra, and X-ray powder diffraction patterns, which may be used for identification. The skilled artisan will appreciate that different polymorphs may be produced, for example, by changing or adjusting the reaction conditions or reagents, used in making the compound. For example, changes in temperature, pressure, or solvent may result in polymophs. In addition, one polymorph may spontaneously convert to another polymorph under certain conditions.
The compounds of the invention inhibit the cathepsin C enzyme and can be useful in the treatment of conditions wherein the underlying pathology is (at least in part) attributable to cathepsin C involvement or in conditions wherein cathepsin C inhibition offers some clinical benefit even though the underlying pathology is not (even in part) attributable to cathepsin C involvement. Examples of such conditions include COPD, rheumatoid arthritis, osteoarthritis, asthma, and multiple sclerosis. Accordingly, in another aspect the invention is directed to methods of treating such conditions.
The methods of treatment of the invention comprise administering a safe and effective amount of a compound of the invention to a patient in need thereof.
As used herein, “treatment” in reference to a condition means: (1) the amelioration or prevention of the condition being treated or one or more of the biological manifestations of the condition being treated, (2) the interference with (a) one or more points in the biological cascade that leads to or is responsible for the condition being treated or (b) one or more of the biological manifestations of the condition being treated, or (3) the alleviation of one or more of the symptoms or effects associated with the condition being treated.
As indicated above, “treatment” of a condition includes prevention of the condition. The skilled artisan will appreciate that “prevention” is not an absolute term. In medicine, “prevention” is understood to refer to the prophylactic administration of a drug to substantially diminish the likelihood or severity of a condition or biological manifestation thereof, or to delay the onset of such condition or biological manifestation thereof.
As used herein, “safe and effective amount” and “therapeutically effective amount” in reference to a compound of formula I, or a pharmaceutically acceptable salt of it, means an amount of sufficient to significantly induce a positive modification in the condition to be treated but low enough to avoid serious side effects at a reasonable benefit/risk ratio within the scope of sound medical judgment. A safe and effective amount of a compound of the invention will vary with the particular compound chosen, e.g. consider the potency, efficacy, and half-life of the compound; the route of administration chosen; the condition being treated; the severity of the condition being treated; the age, size, weight, and physical condition of the patient being treated; the medical history of the patient to be treated; the duration of the treatment; the nature of concurrent therapy; the desired therapeutic effect; and like factors, but can nevertheless be routinely determined by the skilled artisan.
As used herein, “patient” refers to a human or animal.
The compounds of the invention may be administered by any suitable route of administration, including both systemic administration and topical administration. Systemic administration includes oral administration, parenteral administration, transdermal administration, rectal administration, and administration by inhalation. Parenteral administration refers to routes of administration other than enteral, transdermal, or by inhalation, and is typically by injection or infusion. Parenteral administration includes intravenous, intramuscular, and subcutaneous injection or infusion. Inhalation refers to administration into the patient's lungs whether inhaled through the mouth or through the nasal passages. Topical administration includes application to the skin as well as intraocular, otic, intravaginal, and intranasal administration.
The compounds of the invention may be administered once or according to a dosing regimen wherein a number of doses are administered at varying intervals of time for a given period of time. For example, doses may be administered one, two, three, or four times per day. Doses may be administered until the desired therapeutic effect is achieved or indefinitely to maintain the desired therapeutic effect. Suitable dosing regimens for a compound of the invention depend on the pharmacokinetic properties of that compound, such as absorption, distribution, and half-life, which can be determined by the skilled artisan. In addition, suitable dosing regimens, including the amount administered and the duration such regimens are administered, for a compound of the invention depend on the condition being treated, the severity of the condition being treated, the age and physical condition of the patient being treated, the medical history of the patient to be treated, the nature of concurrent therapy, the particular route of administration chosen, the desired therapeutic effect, and like factors within the knowledge and expertise of the skilled artisan. It will be further understood by such skilled artisans that suitable dosing regimens may require adjustment given an individual patient's response to the dosing regimen or over time as individual patient needs change. Typical daily dosages range from 10 mg to 1000 mg.
The compounds of the invention will normally, but not necessarily, be formulated into a pharmaceutical composition prior to administration to a patient. Accordingly, in another aspect the invention is directed to pharmaceutical compositions comprising a compound of the invention and a pharmaceutically-acceptable excipient.
The pharmaceutical compositions of the invention may be prepared and packaged in bulk form wherein a safe and effective amount of a compound of the invention can be extracted and then given to the patient such as with powders, syrups, and solutions for injection. Alternatively, the pharmaceutical compositions of the invention may be prepared and packaged in unit dosage form wherein each physically discrete unit contains a safe and effective amount of a compound of the invention. When prepared in unit dosage form, the pharmaceutical compositions of the invention typically contain from 1 mg to 1000 mg.
The pharmaceutical compositions of the invention typically contain one compound of the invention. However, in certain embodiments, the pharmaceutical compositions of the invention contain more than one compound of the invention. For example, in certain embodiments the pharmaceutical compositions of the invention contain two compounds of the invention. In addition, the pharmaceutical compositions of the invention may optionally further comprise one or more additional pharmaceutically active compounds. Conversely, the pharmaceutical compositions of the invention typically contain more than one pharmaceutically-acceptable excipient. However, in certain embodiments, the pharmaceutical compositions of the invention contain one pharmaceutically-acceptable excipient.
As used herein, “pharmaceutically-acceptable excipient” means a material, composition or vehicle involved in giving form or consistency to the composition and which is safe when administered to a patient. Each excipient must be compatible with the other ingredients of the pharmaceutical composition when commingled such that interactions which would substantially reduce the efficacy of the compound of the invention when administered to a patient and interactions which would result in pharmaceutical compositions that are not pharmaceutically acceptable are avoided. In addition, each excipient must of course be of sufficiently high purity to render it pharmaceutically-acceptable.
The compounds of the invention and the pharmaceutically-acceptable excepient or excepients will typically be formulated into a dosage form adapted for administration to the patient by the desired route of administration. For example, dosage forms include those adapted for (1) oral administration such as tablets, capsules, caplets, pills, troches, powders, syrups, elixers, suspensions, solutions, emulsions, sachets, and cachets; (2) parenteral administration such as sterile solutions, suspensions, and powders for reconstitution; (3) transdermal administration such as transdermal patches; (4) rectal administration such as suppositories; (5) inhalation such as aerosols and solutions; and (6) topical administration such as creams, ointments, lotions, solutions, pastes, sprays, foams, and gels.
Suitable pharmaceutically-acceptable excipients will vary depending upon the particular dosage form chosen. In addition, suitable pharmaceutically-acceptable excipients may be chosen for a particular function that they may serve in the composition. For example, certain pharmaceutically-acceptable excipients may be chosen for their ability to facilitate the production of uniform dosage forms. Certain pharmaceutically-acceptable excipients may be chosen for their ability to facilitate the production of stable dosage forms. Certain pharmaceutically-acceptable excipients may be chosen for their ability to facilitate the carrying or transporting the compound or compounds of the invention once administered to the patient from one organ, or portion of the body, to another organ, or portion of the body. Certain pharmaceutically-acceptable excipients may be chosen for their ability to enhance patient compliance.
Suitable pharmaceutically-acceptable excipients include the following types of excipients: Diluents, fillers, binders, disintegrants, lubricants, glidants, granulating agents, coating agents, wetting agents, solvents, co-solvents, suspending agents, emulsifiers, sweetners, flavoring agents, flavor masking agents, coloring agents, anticaking agents, hemectants, chelating agents, plasticizers, viscosity increasing agents, antioxidants, preservatives, stabilizers, surfactants, and buffering agents. The skilled artisan will appreciate that certain pharmaceutically-acceptable excipients may serve more than one function and may serve alternative functions depending on how much of the excipient is present in the formulation and what other ingredients are present in the formulation.
Skilled artisans possess the knowledge and skill in the art to enable them to select suitable pharmaceutically-acceptable excipients in appropriate amounts for use in the invention. In addition, there are a number of resources that are available to the skilled artisan which describe pharmaceutically-acceptable excipients and may be useful in selecting suitable pharmaceutically-acceptable excipients. Examples include Remington's Pharmaceutical Sciences (Mack Publishing Company), The Handbook of Pharmaceutical Additives (Gower Publishing Limited), and The Handbook of Pharmaceutical Excipients (the American Pharmaceutical Association and the Pharmaceutical Press).
The pharmaceutical compositions of the invention are prepared using techniques and methods known to those skilled in the art. Some of the methods commonly used in the art are described in Remington's Pharmaceutical Sciences (Mack Publishing Company).
In one aspect, the invention is directed to a solid oral dosage form such as a tablet or capsule comprising a safe and effective amount of a compound of the invention and a diluent or filler. Suitable diluents and fillers include lactose, sucrose, dextrose, mannitol, sorbitol, starch (e.g. corn starch, potato starch, and pre-gelatinized starch), cellulose and its derivatives (e.g. microcrystalline cellulose), calcium sulfate, and dibasic calcium phosphate. The oral solid dosage form may further comprise a binder. Suitable binders include starch (e.g. corn starch, potato starch, and pre-gelatinized starch), gelatin, acacia, sodium alginate, alginic acid, tragacanth, guar gum, povidone, and cellulose and its derivatives (e.g. microcrystalline cellulose). The oral solid dosage form may further comprise a disintegrant. Suitable disintegrants include crospovidone, sodium starch glycolate, croscarmelose, alginic acid, and sodium carboxymethyl cellulose. The oral solid dosage form may further comprise a lubricant. Suitable lubricants include stearic acid, magnesium stearate, calcium stearate, and talc.
In another aspect, the invention is directed to a dosage form adapted for administration to a patient by inhalation. For example, the compound of the invention may be inhaled into the lungs as a dry powder, an aerosol, a suspension, or a solution.
Dry powder compositions for delivery to the lung by inhalation typically comprise a compound of the invention as a finely divided powder together with one or more pharmaceutically-acceptable excipients as finely divided powders. Pharmaceutically-acceptable excipients particularly suited for use in dry powders are known to those skilled in the art and include lactose, starch, mannitol, and mono-, di-, and polysaccharides.
The dry powder may be administered to the patient via a reservoir dry powder inhaler (RDPI) having a reservoir suitable for storing multiple (un-metered doses) of medicament in dry powder form. RDPIs typically include a means for metering each medicament dose from the reservoir to a delivery position. For example, the metering means may comprise a metering cup, which is movable from a first position where the cup may be filled with medicament from the reservoir to a second position where the metered medicament dose is made available to the patient for inhalation.
Alternatively, the dry powder may be presented in capsules (e.g. gelatin or plastic), cartridges, or blister packs for use in a multi-dose dry powder inhaler (MDPI). MDPIs are inhalers wherein the medicament is comprised within a multi-dose pack containing (or otherwise carrying) multiple defined doses (or parts thereof) of medicament. When the dry powder is presented as a blister pack, it comprises multiple blisters for containment of the medicament in dry powder form. The blisters are typically arranged in regular fashion for ease of release of the medicament therefrom. For example, the blisters may be arranged in a generally circular fashion on a disc-form blister pack, or the blisters may be elongate in form, for example comprising a strip or a tape. Each capsule, cartridge, or blister may, for example, contain between 20 μg-10 mg of the compound of the invention.
Aerosols may be formed by suspending or dissolving a compound of the invention in a liquified propellant. Suitable propellants include halocarbons, hydrocarbons, and other liquified gases. Representative propellants include: trichlorofluoromethane (propellant 11), dichlorofluoromethane (propellant 12), dichlorotetrafluoroethane (propellant 114), tetrafluoroethane (HFA-134a), 1,1-difluoroethane (HFA-152a), difluoromethane (HFA-32), pentafluoroethane (HFA-12), heptafluoropropane (HFA-227a), perfluoropropane, perfluorobutane, perfluoropentane, butane, isobutane, and pentane. Aerosols comprising a compound of the invention will typically be administered to a patient via a metered dose inhaler (MDI). Such devices are known to those skilled in the art.
The aerosol may contain additional pharmaceutically-acceptable excipients typically used with multiple dose inhalers such as surfactants, lubricants, cosolvents and other excipients to improve the physical stability of the formulation, to improve valve performance, to improve solubility, or to improve taste.
Suspensions and solutions comprising a compound of the invention may also be administered to a patient via a nebulizer. The solvent or suspension agent utilized for nebulization may be any pharmaceutically-acceptable liquid such as water, aqueous saline, alcohols or glycols, e.g., ethanol, isopropylalcohol, glycerol, propylene glycol, polyethylene glycol, etc. or mixtures thereof. Saline solutions utilize salts which display little or no pharmacological activity after administration. Both organic salts, such as alkali metal or ammonium halogen salts, e.g., sodium chloride, potassium chloride or organic salts, such as potassium, sodium and ammonium salts or organic acids, e.g., ascorbic acid, citric acid, acetic acid, tartaric acid, etc. may be used for this purpose.
Other pharmaceutically-acceptable excipients may be added to the suspension or solution. The compound of the invention may be stabilized by the addition of an inorganic acid, e.g., hydrochloric acid, nitric acid, sulphuric acid and/or phosphoric acid; an organic acid, e.g., ascorbic acid, citric acid, acetic acid, and tartaric acid, etc., a complexing agent such as EDTA or citric acid and salts thereof, or an antioxidant such as antioxidant such as vitamin E or ascorbic acid. These may be used alone or together to stabilize the compound of the invention. Preservatives may be added such as benzalkonium chloride or benzoic acid and salts thereof. Surfactant may be added particularly to improve the physical stability of suspensions. These include lecithin, disodium dioctylsulphosuccinate, oleic acid and sorbitan esters.
aq. Aqueous
atm atmosphere
BOC tert-butyloxycarbonyl
DCM/CH2Cl2 dichloromethane
DEAD Diethylazodicarboxylate
DMAP Dimethylaminopyridine
DIPEA/DIEA Di-isopropylethylamine
DMF Dimethylformamide
DPPA Diphenylphosphoryl azide
EA/EtOAc Ethyl acetate
ESI Electrospray ionization
eq. Equivalent
HPLC High pressure liquid chromatography
LC-MS Liquid chromatography-Mass spectrometry
MDAP Mass Directed Auto Prep
Me Methyl
min Minutes
ml or mL milliliter
Ph Phenyl
PS Polymer-supported
rt Room temperature
sat or sat. Saturated
SPE Solid phase extraction
TBAF tetra-Butylammonium fluroride
TBS t-Butyldimethyl silyl
TBS-Cl t-Butyldimethyl silyl chloride
TEA Triethylamine
TFA Trifluoroacetic acid
THF Tetrahydrofuran
UV Ultraviolet
The compounds of this invention may be made by a variety of methods, including standard chemistry. Any previously defined variable will continue to have the previously defined meaning unless otherwise indicated. Illustrative general synthetic methods are set out below and then specific compounds of the invention as prepared are given in the examples.
The compounds of formula (IA) and (IB) may be obtained by using synthetic procedures illustrated in the Schemes below or by drawing on the knowledge of a skilled organic chemist. The synthesis provided in these Schemes are applicable for producing compounds of the invention having a variety of different R1 groups employing appropriate precursors, which are suitable protected if needs be, to achieve compatibility with the reactions outlined herein. Subsequent deprotection, where needs be, and then affords compounds of the nature generally disclosed. While the Schemes are shown with compounds only of formula (IA) or (IB), they are illustrative of processes that may be used to make the compounds of the invention.
Compounds names were generated using the software naming program ACD/Name Pro V6.02 available from Advanced Chemistry Development, Inc., 110 Yonge Street, 14th Floor, Toronto, Ontario, Canada, M5C 1T4 (http://www.acdlabs.com/).
As shown in Scheme 1, the compounds of the formula (IA) can be prepared in a multi-step sequence from the commercially available 1,1-dimethylethyl (3R)-3-amino-1-pyrrolidinecarboxylate 1. Treatment of 1 with a suitable sulfonyl chloride (R1)nC6H4SO2Cl gives the derivative 2. Subsequent removal of the BOC protecting group with an acid reagent such as 4N HCl in dioxane followed by treatment with cyanogen bromide results in the formation of the desired compounds 3 of formula (IA).
Derivatives substituted with an amide group, a urea group or an aminoalkyl group at the meta-position of the phenyl ring can be prepared following the reaction sequence depicted in Scheme 2. The nitro intermediate 4, which can be prepared following procedures analogous to that described in Scheme 1, can be reduced to the corresponding aniline 5 following procedures well known in the art such as hydrogenation in the presence of a metal catalyst. Coupling of compound 5 with a suitable reagent such as an acyl chloride or an isocyanate give the protected intermediates 7 and 6, which can be respectively transformed, following a two-step cyanation procedure analogous to that presented in Scheme 1, into the amide 10 or the urea 9 of general formula (IA). A reductive amination can also be performed on 5 to give the N-alkylated intermediate 8. A subsequent a two-step cyanation procedure analogous to that presented in Scheme 1 gives compound 11 of formula (I).
Purifications and analyses of materials were carried out using the following instruments:
Automated preparatory HPLC purifications were conducted with a Gilson® semi-preparative HPLC system under the following conditions:
Column: 75×33 mm I. D., S-5 um, 12 nm
Flow rate: 30 mL/min
Injection Volume: 0.800 mL
Room temperature
The eluent was a mixture composed of solvents A and B:
Solvent A: 0.1% trifluoroacetic acid in water
Solvent B: 0.1% trifluoroacetic acid in acetonitrile
The Mass-Directed Auto Prep HPLC (MDAP) purifications were conducted with an Agilent preparatory HPLC-MS system under the following conditions:
Column: ZORBAX Eclipse XDB-C18 (21.2×50 mm)
Flow rate: 20 mL/min
Injection volume: 900 uL
Temperature: 30° C.
absorption wavelength: 230 nm
The eluent was a mixture composed of solvents A and B:
Solvent A: 0.1% trifluoroacetic acid in water
Solvent B: 0.1% trifluoroacetic acid in acetonitrile
The automated flash chromatography purifications were conducted with a CombiFlash® Companion® personal flash chromatography system under the following conditions:
Silica cartridge:
Size, 4, 12, 40, 80 or 120 g depending on the amount of material to be purified
Flow rate: Between 4 and 85 mL/min
Room temperature
The eluent was a mixture composed of solvents A and B:
Solvent A: Hexane
Solvent B: Ethyl acetate
Unless specified otherwise, the following abbreviations were used for frequently used reagents:
BrCN: A 3 N BrCN solution DCM (Aldrich®, cat #:341894)
PS-trisamine: A tris-(2-aminoethyl)amine polystyrene resin (Argonaut®, p/n 800230)
4 N HCl: A 4 N HCl solution in 1,4-dioxane (Aldrich cat #345547)
All solvents used herein are of the highest available purity and all reactions are run under anhydrous conditions under an air atmosphere unless otherwise indicated.
The invention will now be described by reference to the following Examples which are merely illustrative and are not to be construed as a limitation of the scope of the present invention. All temperatures are given in ° C.
To 5-methyl-2-(methyloxy)benzenesulfonyl chloride (0.1101 g, 0.50 mmol) and triethylamine (0.275 ml, 1.97 mmol) in DCM (2 mL) was added 1,1-dimethylethyl (3R)-3-amino-1-pyrrolidinecarboxylate (0.080 ml, 0.471 mmol). The reaction mixture was stirred at room temperature overnight. Water (1.5 ml) was added to the reaction mixture with stirring. The mixture was then diluted with DCM (2 ml) and water (1.5 ml) and put through a phase separator to dry. Then 4N HCl in 1,4-dioxane (2.0 ml) was added. After 6 hours, the reaction was blown down to dryness. The dry material was then diluted with DCM (10 ml), and mixed with DIEA (0.45 mL, 2.58 mmol) and BrCN (0.40 mL, 1.2 mmol). The resultant mixture was stirred at room temperature overnight. The solvent was evaporated under vacuum and the solid purified by preparatory HPLC (without TFA) to afford the title compound (0.0698 g). LC-MS: m/z, 296 (M+H), rt 1.48 min.
To 2-methylbenzenesulfonyl chloride (0.096 g, 0.50 mmol) and triethylamine (0.275 ml, 1.97 mmol) in DCM (2 mL) was added 1,1-dimethylethyl (3R)-3-amino-1-pyrrolidinecarboxylate (0.080 ml, 0.471 mmol). The reaction mixture was stirred at room temperature overnight. Water (1.5 ml) was added to the reaction mixture with stirring. The mixture was then diluted with DCM (2 ml) and water (1.5 ml) and put through a phase separator to dry. Then 4N HCl in 1,4-dioxane (2.0 ml) was added. After 6 hours, the reaction was blown down to dryness. The dry materials was then diluted with DCM (10 ml), and mixed with DIEA (0.45 mL, 2.58 mmol) and BrCN (0.40 mL, 1.2 mmol). The resultant mixture was stirred at room temperature overnight. The solvent was evaporated under vacuum and the solid purified by preparatory HPLC (without TFA) to afford the title compound (0.0614 g). LC-MS: m/z, 266 (M+H), rt 1.36 min.
To 3,5-bis(trifluoromethyl)benzenesulfonyl chloride (0.1569 g, 0.50 mmol) and triethylamine (0.275 ml, 1.97 mmol) in DCM (2 mL) was added 1,1-dimethylethyl (3R)-3-amino-1-pyrrolidinecarboxylate (0.080 ml, 0.471 mmol). The reaction mixture was stirred at room temperature overnight. Water (1.5 ml) was added with stirring. The mixture was then diluted with DCM (2 ml) and water (1.5 ml) and put through a phase separator to dry. Then 4N HCl in 1,4-dioxane (2.0 ml) was added. After 6 hours, the reaction was blown down to dryness. The dry material was then diluted with DCM (10 ml), and mixed with DIEA (0.45 mL, 2.58 mmol) and BrCN (0.40 mL, 1.2 mmol). The resultant mixture was stirred at room temperature overnight. The solvent was evaporated under vacuum and the solid purified by preparatory HPLC (without TFA) to afford the title compound (0.0846 g). LC-MS: m/z, 388 (M+H), rt 1.95 min.
To 2,5-bis(methyloxy)benzenesulfonyl chloride (0.119 g, 0.50 mmol) and triethylamine (0.275 ml, 1.97 mmol) in DCM (2 mL) was added 1,1-dimethylethyl (3R)-3-amino-1-pyrrolidinecarboxylate (0.080 ml, 0.471 mmol). The reaction mixture was stirred at room temperature overnight. Water (1.5 ml) was added to the reaction mixture with stirring. The mixture was then diluted with DCM (2 ml) and water (1.5 ml) and put through a phase separator to dry it. Then 4N HCl in 1,4-dioxane (2.0 ml) was added. After 6 hours, the mixture was blown down to dryness. The dry material was then diluted with DCM (10 ml), and mixed with DIEA (0.45 mL, 2.58 mmol) and BrCN (0.40 mL, 1.2 mmol). The resultant mixture was stirred at room temperature overnight. The solvent was evaporated under vacuum and the solid purified by preparatory HPLC (without TFA) to afford the title compound (0.0757 g). LC-MS: m/z, 312 (M+H), rt 1.45 min.
To 5-chloro-2-(methyloxy)benzenesulfonyl chloride (0.1212 g, 0.50 mmol) and triethylamine (0.275 ml, 1.97 mmol) in DCM (2 mL) was added 1,1-dimethylethyl (3R)-3-amino-1-pyrrolidinecarboxylate (0.080 ml, 0.471 mmol). The reaction mixture was stirred at room temperature overnight. Water (1.5 ml) was added to the mixture with stirring. The mixture was then diluted with DCM (2 ml) and water (1.5 ml) and put through a phase separator to dry. Then 4N HCl in 1,4-dioxane (2.0 ml) was added. After 6 hours, the mixture was blown down to dryness. It was then diluted with DCM (10 ml), and mixed with DIEA (0.45 mL, 2.58 mmol) and BrCN (0.40 mL, 1.2 mmol). The resultant mixture was stirred at room temperature overnight. The solvent was evaporated under vacuum and the solid purified by preparatory HPLC (without TFA) to afford the title compound (0.0763 g). LC-MS: m/z, 316 (M+H), rt 1.62 min.
To 3-cyanobenzenesulfonyl chloride (0.1019 g, 0.50 mmol) and triethylamine (0.275 ml, 1.97 mmol) in DCM (2 mL) was added 1,1-dimethylethyl (3R)-3-amino-1-pyrrolidinecarboxylate (0.080 ml, 0.471 mmol). The reaction mixture was stirred at room temperature overnight. Water (1.5 ml) was added to the reaction mixture with stirring. The mixture was then diluted with DCM (2 ml) and water (1.5 ml) and put through a phase separator to dry. Then 4N HCl in 1,4-dioxane (2.0 ml) was added. After 6 hours, the reaction was blown down to dryness. The dry material was then diluted with DCM (10 ml), and mixed with DIEA (0.45 mL, 2.58 mmol) and BrCN (0.40 mL, 1.2 mmol). The resultant mixture was stirred at room temperature overnight. The solvent was evaporated under vacuum and the solid purified by preparatory HPLC (without TFA) to afford the title compound (0.0665 g). LC-MS: m/z, 277 (M+H), rt 1.40 min.
To a chilled solution of 3-chlorobenzenesulfonyl chloride (0.07 ml, 0.50 mmol) and triethylamine (0.3 ml, 2.15 mmol) in DCM (2 mL) was added 1,1-dimethylethyl (3R)-3-amino-1-pyrrolidinecarboxylate (0.080 ml, 0.471 mmol). The reaction mixture was stirred at room temperature overnight. Water (1.5 ml) was added to the reaction mixture with stirring. The reaction was then diluted with DCM (2 ml) and water (1.5 ml) and put through a phase separator to dry. Then 4N HCl in 1,4-dioxane (2.0 ml) was added. After 1.5 hours, the reaction was blown down to dryness. The dry material was then diluted with DCM (8 ml), and mixed with DIEA (0.45 mL, 2.58 mmol) and BrCN (0.40 mL, 1.2 mmol). The resultant mixture was stirred at room temperature overnight. The solvent was evaporated under vacuum and the solid purified by preparatory HPLC (without TFA) to afford the title compound (0.0326 g). LC-MS: m/z, 286 (M+H), rt 1.56 min.
To a chilled solution of 2,5-bis(trifluoromethyl)benzenesulfonyl chloride (0.163 g, 0.52 mmol) and triethylamine (0.3 ml, 2.15 mmol) in DCM (2 mL) was added 1,1-dimethylethyl (3R)-3-amino-1-pyrrolidinecarboxylate (0.080 ml, 0.471 mmol). The reaction mixture was stirred at room temperature overnight. Water (1.5 ml) was added to the reaction mixture with stirring. The reaction was then diluted with DCM (2 ml) and water (1.5 ml) and put through a phase separator to dry. Then 4N HCl in 1,4-dioxane (2.0 ml) was added. After 1.5 hours, the mixture was blown down to dryness. The dry material was then diluted with DCM (8 ml), and mixed with DIEA (0.45 mL, 2.58 mmol) and BrCN (0.40 mL, 1.2 mmol). The resultant mixture was stirred at room temperature overnight. The solvent was evaporated under vacuum and the solid purified by preparatory HPLC (without TFA) to afford the title compound (0.0637 g). LC-MS: m/z, 388 (M+H), rt 1.89 min.
To a chilled solution of 2-fluoro-5-methylbenzenesulfonyl chloride (0.114 g, 0.546 mmol) and triethylamine (0.3 ml, 2.15 mmol) in DCM (2 mL) was added 1,1-dimethylethyl (3R)-3-amino-1-pyrrolidinecarboxylate (0.080 ml, 0.471 mmol). The reaction mixture was stirred at room temperature overnight. Water (1.5 ml) was added to the reaction mixture with stirring. The reaction was then diluted with DCM (2 ml) and water (1.5 ml) and put through a phase separator to dry. Then 4N HCl in 1,4-dioxane (2.0 ml) was added. After 1.5 hours, the reaction was blown down to dryness. The dry material was then diluted with DCM (8 ml), and mixed with DIEA (0.45 mL, 2.58 mmol) and BrCN (0.40 mL, 1.2 mmol). The resultant mixture was stirred at room temperature overnight. The solvent was evaporated under vacuum and the solid purified by preparatory HPLC (without TFA) to afford the title compound (0.0554 g). LC-MS: m/z, 284 (M+H), rt 1.51 min.
To a chilled solution of 5-chloro-2-fluorobenzenesulfonyl chloride (0.125 g, 0.546 mmol) and triethylamine (0.3 ml, 2.15 mmol) in DCM (2 mL) was added 1,1-dimethylethyl (3R)-3-amino-1-pyrrolidinecarboxylate (0.080 ml, 0.471 mmol). The reaction mixture was stirred at room temperature overnight. Water (1.5 ml) was added to the reaction mixture with stirring. The mixture was diluted with DCM (2 ml) and water (1.5 ml) and put through a phase separator to dry. Then 4N HCl in 1,4-dioxane (2.0 ml) was added. After 1.5 hours, the reaction was blown down to dryness. The dry material was then diluted with DCM (8 ml), and mixed with DIEA (0.45 mL, 2.58 mmol) and BrCN (0.40 mL, 1.2 mmol). The resultant mixture was stirred at room temperature overnight. The solvent was evaporated under vacuum and the solid purified by preparatory HPLC (without TFA) to afford the title compound (0.0671 g). LC-MS: m/z, 304 (M+H), rt 1.6 min.
To a chilled (0° C.) solution of 1,1-dimethylethyl (3R)-3-amino-1-pyrrolidinecarboxylate (0.095 ml, 0.0560 mmol) in DMF (1 ml) was added triethylamine (0.310 ml, 2.22 mmol) and 3-bromobenzenesulfonyl chloride (0.290 g, 1.14 mmol). The reaction mixture was stirred at room temperature overnight. The reaction was diluted with EtOAc, washed with water (2×) then brine, dried MgSO4, filtered and concentrated. Then 4N HCl in 1,4-dioxane (5.0 ml) was added. After stirring at room temperature overnight, the solvent was evaporated. The dry material was then diluted with DCM (10 ml), and mixed with DIEA (0.2 mL, 1.15 mmol) and BrCN (0.40 mL, 1.2 mmol). The resultant mixture was stirred at room temperature overnight. The solvent was evaporated and the solid purified by preparatory HPLC (without TFA) to afford the title compound (0.0916 g). LC-MS: m/z, 330 (M+H), rt 1.70 min.
To a chilled (0° C.) solution of 1,1-dimethylethyl (3R)-3-amino-1-pyrrolidinecarboxylate (0.095 ml, 0.0560 mmol) in DMF (1 ml) was added triethylamine (0.310 ml, 2.22 mmol) and 2-bromobenzenesulfonyl chloride (0.291 g, 1.14 mmol). The reaction mixture was stirred at room temperature overnight. The reaction was diluted with EtOAc, washed with water (2×) then brine, dried MgSO4, filtered and concentrated. Then 4N HCl in 1,4-dioxane (5.0 ml) was added. After stirring at room temperature overnight, the solvent was evaporated. The resulting solid was then diluted with DCM (10 ml), and mixed with DIEA (0.2 mL, 1.15 mmol) and BrCN (0.40 mL, 1.2 mmol). The resultant mixture was stirred at room temperature overnight. The solvent was evaporated and the solid purified by preparatory HPLC (without TFA) to afford the title compound (0.0304 g). LC-MS: m/z, 330 (M+H), rt 1.59 min.
To a chilled (0° C.) solution of 1,1-dimethylethyl (3R)-3-amino-1-pyrrolidinecarboxylate (0.085 ml, 0.0501 mmol) in THF (1 ml) was added triethylamine (0.275 ml, 1.97 mmol) and 2,3,5,6-tetramethylbenzenesulfonyl chloride (0.117 g, 0.50 mmol). The reaction mixture was stirred at room temperature overnight. The reaction was diluted with water, extracted with DCM and concentrated. Then 4N HCl in 1,4-dioxane (3.0 ml) was added. After stirring at room temperature overnight, the solvent was evaporated. The mixture was then diluted with DCM (10 ml), and mixed with DIEA (0.2 mL, 1.15 mmol) and BrCN (0.40 mL, 1.2 mmol). The resultant mixture was stirred at room temperature overnight. The solvent was evaporated under vacuum and the solid purified by preparatory HPLC (without TFA) to afford the title compound (0.0647 g). LC-MS: m/z, 308 (M+H), rt 1.84 min.
To a chilled (0° C.) solution of 1,1-dimethylethyl (3R)-3-amino-1-pyrrolidinecarboxylate (0.085 ml, 0.0501 mmol) in THF (1 ml) was added triethylamine (0.275 ml, 1.97 mmol) and 3-(chlorosulfonyl)benzoic acid (0.111 g, 0.50 mmol). The reaction mixture was stirred at room temperature overnight. The reaction was diluted with water, extracted with DCM and concentrated. Then 4N HCl in 1,4-dioxane (3.0 ml) was added. After stirring at room temperature overnight, the solvent was evaporated. The mixture was diluted with DCM (10 ml), and mixed with DIEA (0.2 mL, 1.15 mmol) and BrCN (0.40 mL, 1.2 mmol). The resultant mixture was stirred at room temperature overnight. The solvent was evaporated under vacuum. The solid materials was then re-exposed to the above conditions. The mixture was diluted with DCM (10 ml), and mixed with DIEA (0.1 mL, 0.58 mmol) and BrCN (0.40 mL, 1.2 mmol). The resultant mixture was stirred at room temperature overnight. The solvent was evaporated under vacuum and the solid purified by preparatory HPLC (without TFA) to afford the title compound (0.0086 g). LC-MS: m/z, 389 (M+H), rt 1.44 min.
To a chilled (0° C.) solution of 1,1-dimethylethyl (3R)-3-amino-1-pyrrolidinecarboxylate (0.085 ml, 0.0501 mmol) in THF (1 ml) was added triethylamine (0.275 ml, 1.97 mmol) and 2-chlorobenzenesulfonyl chloride (0.1062 g, 0.50 mmol). The reaction mixture was stirred at room temperature overnight. It was diluted with water, extracted with DCM and concentrated. Then 4N HCl in 1,4-dioxane (3.0 ml) was added. After stirring at room temperature overnight, the solvent was evaporated. The residue was then diluted with DCM (10 ml), and mixed with DIEA (0.2 mL, 1.15 mmol) and BrCN (0.40 mL, 1.2 mmol). The solution was stirred at room temperature overnight. The solvent was evaporated under vacuum and the solid purified by preparatory HPLC (without TFA) to afford the title compound (0.0834 g). LC-MS: m/z, 286 (M+H), rt 1.43 min.
To a chilled (0° C.) solution of 1,1-dimethylethyl (3R)-3-amino-1-pyrrolidinecarboxylate (0.085 ml, 0.0501 mmol) in THF (1 ml) was added triethylamine (0.275 ml, 1.97 mmol) and 3,5-dichlorobenzenesulfonyl chloride (0.124 g, 0.50 mmol). The reaction mixture was stirred at room temperature overnight. It was diluted with water, extracted with DCM and concentrated. Then 4N HCl in 1,4-dioxane (3.0 ml) was added. After stirring at room temperature overnight, the solvent was evaporated. The residue was then diluted with DCM (10 ml), and mixed with DIEA (0.2 mL, 1.15 mmol) and BrCN (0.40 mL, 1.2 mmol). The resultant mixture was stirred at room temperature overnight. The solvent was evaporated under vacuum and the solid purified by preparatory HPLC (without TFA) to afford the title compound (0.0207 g). LC-MS: m/z, 319 (M+H), rt 1.83 min.
To a chilled (0° C.) solution of 1,1-dimethylethyl (3R)-3-amino-1-pyrrolidinecarboxylate (0.085 ml, 0.0501 mmol) in THF (1 ml) was added triethylamine (0.275 ml, 1.97 mmol) and 3-chloro-2-methylbenzenesulfonyl chloride (0.1131 g, 0.50 mmol). The reaction mixture was stirred at room temperature overnight. It was diluted with water, extracted with DCM and concentrated. To the reaction was added 4N HCl in 1,4-dioxane (3.0 ml). After stirring at room temperature overnight, the solvent was evaporated. The residue was diluted with DCM (10 ml), and mixed with DIEA (0.2 mL, 1.15 mmol) and BrCN (0.40 mL, 1.2 mmol). The resultant mixture was stirred at room temperature overnight. The solvent was evaporated under vacuum. The residue was then re-exposed to the above conditions. The resulting material was diluted with DCM (10 ml), and mixed with DIEA (0.1 mL, 0.58 mmol) and BrCN (0.40 mL, 1.2 mmol) and stirred at room temperature overnight. The solvent was evaporated under vacuum and the solid purified by preparatory HPLC (without TFA) to afford the title compound (0.0567 g). LC-MS: m/z, 300 (M+H), rt 1.73 min.
To a chilled (0° C.) solution of 1,1-dimethylethyl (3R)-3-amino-1-pyrrolidinecarboxylate (0.085 ml, 0.0501 mmol) in THF (1 ml) was added triethylamine (0.275 ml, 1.97 mmol) and 2,6-dichlorobenzenesulfonyl chloride (0.1235 g, 0.50 mmol). The reaction mixture was stirred at room temperature overnight, then diluted with water, extracted with DCM and concentrated. To the residue was added 4N HCl in 1,4-dioxane (3.0 ml). After stirring at room temperature overnight, the solvent was evaporated, the residue diluted with DCM (10 ml), and mixed with DIEA (0.2 mL, 1.15 mmol) and BrCN (0.40 mL, 1.2 mmol). The resultant mixture was stirred at room temperature overnight. The solvent was evaporated under vacuum and the solid purified by preparatory HPLC (without TFA) to afford the title compound (0.0412 g). LC-MS: m/z, 320 (M+H), rt 1.66 min.
To a chilled (0° C.) solution of 1,1-dimethylethyl (3R)-3-amino-1-pyrrolidinecarboxylate (0.085 ml, 0.0501 mmol) in THF (1 ml) was added triethylamine (0.275 ml, 1.97 mmol) and 2-cyanobenzenesulfonyl chloride (0.101 g, 0.50 mmol). The reaction mixture was stirred at room temperature overnight, diluted with water, extracted with DCM and concentrated. Then 4N HCl in 1,4-dioxane (3.0 ml) was added. After stirring at room temperature overnight, the solvent was evaporated the residue taken up in DCM (10 ml), and mixed with DIEA (0.2 mL, 1.15 mmol) and BrCN (0.40 mL, 1.2 mmol). The resultant mixture was stirred at room temperature overnight. The solvent was evaporated under vacuum and the solid purified by preparatory HPLC (without TFA) to afford the title compound (0.0070 g). LC-MS: m/z, 277 (M+H), rt 1.40 min.
To a chilled (0° C.) solution of 1,1-dimethylethyl (3R)-3-amino-1-pyrrolidinecarboxylate (0.085 ml, 0.0501 mmol) in THF (1 ml) was added triethylamine (0.275 ml, 1.97 mmol) and 5-fluoro-2-methylbenzenesulfonyl chloride (0.1043 g, 0.50 mmol). The reaction mixture was stirred at room temperature overnight, diluted with water, extracted with DCM and concentrated. To the residue was added 4N HCl in 1,4-dioxane (3.0 ml). After stirring at room temperature overnight, the solvent was evaporated, the residue taken up in DCM (10 ml), to which was added DIEA (0.2 mL, 1.15 mmol) and BrCN (0.40 mL, 1.2 mmol). The resultant mixture was stirred at room temperature overnight. The solvent was evaporated under vacuum and the solid purified by preparatory HPLC (without TFA) to afford the title compound (0.0263 g). LC-MS: m/z, 284 (M+H), rt 1.67 min.
To a chilled (0° C.) solution of 1,1-dimethylethyl (3R)-3-amino-1-pyrrolidinecarboxylate (0.085 ml, 0.0501 mmol) in DMF (1 ml) was added triethylamine (0.275 ml, 1.97 mmol) and 2,5-dibromobenzenesulfonyl chloride (0.167 g, 0.50 mmol). The reaction mixture was stirred at room temperature overnight, diluted with water, extracted with DCM and concentrated. To the concentrate was added DCM (15 ml) and 4N HCl in 1,4-dioxane (5.0 ml). After stirring at room temperature overnight, the solvent was evaporated, the residue diluted with DCM (10 ml), and mixed with DIEA (0.2 mL, 1.15 mmol) and BrCN (0.20 mL, 0.6 mmol). The resultant mixture was stirred at room temperature overnight. The solvent was evaporated and purified by preparatory HPLC (without TFA) to afford the title compound (0.069 g). LC-MS: m/z, 408 (M+H), rt 1.72 min.
To a chilled (0° C.) solution of 1,1-dimethylethyl (3R)-3-amino-1-pyrrolidinecarboxylate (0.085 ml, 0.0501 mmol) in DMF (1 ml) was added triethylamine (0.275 ml, 1.97 mmol) and 2,5-dichlorobenzenesulfonyl chloride (0.122 g, 0.50 mmol). The reaction mixture was stirred at room temperature overnight, diluted with water, extracted with DCM and concentrated. Then DCM (15 ml) and 4N HCl in 1,4-dioxane (5.0 ml) were added. After stirring at room temperature overnight, the solvent was evaporated, the residue diluted with DCM (10 ml), and mixed with DIEA (0.2 mL, 1.15 mmol) and BrCN (0.20 mL, 0.6 mmol). The resultant mixture was stirred at room temperature overnight. The solvent was evaporated and the solid purified by preparatory HPLC (without TFA) to afford the title compound (0.0509 g). LC-MS: m/z, 320 (M+H), rt 1.65 min.
To a chilled (0° C.) solution of 1,1-dimethylethyl (3R)-3-amino-1-pyrrolidinecarboxylate (0.085 ml, 0.0501 mmol) in DMF (1 ml) was added triethylamine (0.275 ml, 1.97 mmol) and 3-(trifluoromethyl)benzenesulfonyl chloride (0.1055 g, 0.50 mmol). The reaction mixture was stirred at room temperature overnight, diluted with water, extracted with DCM and concentrated. To the concentrate was added DCM (15 ml) and 4N HCl in 1,4-dioxane (5.0 ml). After stirring at room temperature overnight, the solvent was evaporated, the residue diluted with DCM (10 ml), and mixed with DIEA (0.2 mL, 1.15 mmol) and BrCN (0.20 mL, 0.6 mmol). The resultant mixture was stirred at room temperature overnight. The solvent was evaporated and the solid purified by preparatory HPLC (without TFA) to afford the title compound (0.0408 g). LC-MS: m/z, 320 (M+H), rt 1.66 min.
To a chilled (0° C.) solution of 1,1-dimethylethyl (3R)-3-amino-1-pyrrolidinecarboxylate (0.085 ml, 0.0501 mmol) in DMF (1 ml) was added triethylamine (0.275 ml, 1.97 mmol) and 3-methylbenzenesulfonyl chloride (0.0953 g, 0.50 mmol). The reaction mixture was stirred at room temperature overnight, diluted with water, extracted with DCM and concentrated. To the concentrate was added DCM (15 ml) and 4N HCl in 1,4-dioxane (5.0 ml). After stirring at room temperature overnight, the solvent was evaporated, the residue diluted with DCM (10 ml), and mixed with DIEA (0.2 mL, 1.15 mmol) and BrCN (0.20 mL, 0.6 mmol). The resultant mixture was stirred at room temperature overnight. The solvent was evaporated and the solid purified by preparatory HPLC (without TFA) to afford the title compound (0.0818 g). LC-MS: m/z, 266 (M+H), rt 1.45 min.
To a chilled (0° C.) solution of 1,1-dimethylethyl (3R)-3-amino-1-pyrrolidinecarboxylate (0.085 ml, 0.0501 mmol) in DMF (1 ml) was added triethylamine (0.275 ml, 1.97 mmol) and 2,5-dimethylbenzenesulfonyl chloride (0.1023 g, 0.50 mmol). The reaction mixture was stirred at room temperature overnight, diluted with water, extracted with DCM and concentrated. To the concentrate was added DCM (15 ml) and 4N HCl in 1,4-dioxane (5.0 ml). After stirring at room temperature overnight, the solvent was evaporated, residue diluted with DCM (10 ml), and mixed with DIEA (0.2 mL, 1.15 mmol) and BrCN (0.20 mL, 0.6 mmol). The resultant mixture was stirred at room temperature overnight. The solvent was evaporated and the solid purified by preparatory HPLC (without TFA) to afford the title compound (0.0912 g). LC-MS: m/z, 280 (M+H), rt 1.58 min.
To a chilled (0° C.) solution of 1,1-dimethylethyl (3R)-3-amino-1-pyrrolidinecarboxylate (0.085 ml, 0.0501 mmol) in DMF (1 ml) was added triethylamine (0.275 ml, 1.97 mmol) and 2-chloro-5-(trifluoromethyl)benzenesulfonyl chloride (0.1395 g, 0.50 mmol). The reaction mixture was stirred at room temperature overnight, diluted with water, extracted with DCM and concentrated. To the concentrate was added DCM (15 ml) and 4N HCl in 1,4-dioxane (5.0 ml). After stirring at room temperature overnight, the solvent was evaporated, the residue diluted with DCM (10 ml), and mixed with DIEA (0.2 mL, 1.15 mmol) and BrCN (0.20 mL, 0.6 mmol). The resultant mixture was stirred at room temperature overnight. The solvent was evaporated and the solid purified by preparatory HPLC (without TFA) to afford the title compound (0.0162 g). LC-MS: m/z, 354 (M+H), rt 1.75 min.
To 1,1-dimethylethyl (3R)-3-{[(2-bromo-5-nitrophenyl)sulfonyl]amino}-1-pyrrolidinecarboxylate (0.0733 g, 0.163 mmol) was added 4N HCl in 1,4-dioxane (0.79 ml). After 2 hours, the reaction was evaporated in vacuo to dryness. The residue was then diluted with DCM (2 ml), and mixed with DIEA (0.110 mL, 0.631 mmol) and BrCN (0.108 mL, 0.324 mmol). The resultant mixture was stirred at room temperature overnight. PS-trisamine resin was added to the reaction for 3 hours, then the solution was filtered, concentrated and the resulting solid purified by preparatory HPLC (without TFA) to afford the title compound (0.010 g). LC-MS: m/z, 375 (M+H), rt 1.12 min.
To a solution of 1,1-dimethylethyl (3R)-3-amino-1-pyrrolidinecarboxylate (6.4 g, 0.034 mol) and 2-(methyloxy)-5-nitrobenzenesulfonyl chloride (13.19 g, 0.051 mol) in DCM (80 ml) was added pyridine (3.4 ml, 0.041 mol). The reaction mixture was stirred at room temperature for 24 hours. More pyridine (1.7 ml, 0.020 mol) was added and the reaction was stirred at room temperature for 2 hours followed by the addition of more 2-(methyloxy)-5-nitrobenzenesulfonyl chloride (1.3 g, 5.12 mmol). The reaction was stirred at room temperature for 20 hours, and then purified by flash chromatography (100 g silica cartridge, 0-100% EtOAc/DCM, 60 min) to afford the title compound (8.95 g). Less pure fractions were combined and repurified (same conditions) to afford more of the title compound (4.36 g). LC-MS: m/z, 402 (M+H), rt 3.09 min.
A solution of 1,1-dimethylethyl (3R)-3-({[2-(methyloxy)-5-nitrophenyl]sulfonyl}amino)-1-pyrrolidinecarboxylate (13.31 g, 0.033 mol) in EtOH (250 ml) was added to a flask containing 10% Pd/C (1.4 g) and the reaction mixture was hydrogenated at room temperature for 20 hours. More 10% Pd/C (0.4 g) was added and the reaction was hydrogenated for an additional 20 hours. The catalyst was filtered off under nitrogen, and the filtrate evaporated. The residue was dissolved in DCM and evaporated again and then it was purified by flash chromatography (2×100 g silica cartridge, 0-100% EtOAc/DCM) to afford the title compound (11.4 g). Less pure fractions were combined and repurified (100 g silica cartridge, 0-100% EtOAc/DCM, 60 min) to afford more of the title compound (0.7 g). LC-MS: m/z, 372 (M+H), rt 2.68 min.
To 2-chloro-5-nitrobenzenesulfonyl chloride (9.98 g, 38 mmol) in DCM (75 mL) was added pyridine (3.8 ml, 46.8 mmol) followed by 1,1-dimethylethyl (3R)-3-amino-1-pyrrolidinecarboxylate (7.25 g, 39 mmol). The reaction mixture was stirred at room temperature for 18 hours. The solvent was evaporated to a small volume which was subjected to purification by flash chromatography (silica cartridge, 100% DCM to 50% DCM/50% EtOAc). The product containing fractions were combined and purified further with flash chromatography (silica cartridge, 97% DCM/3% (2M NH4)MeOH) to afford the title compound (6.5 g).
1,1-Dimethylethyl (3R)-3-{[(2-chloro-5-nitrophenyl)sulfonyl]amino}-1-pyrrolidinecarboxylate (0.727 g, 1.791 mmol) in EtOH-EtOAc (1:1, 50 ml) was hydrogenated (room temp, 1 atm) over 5% platinum on carbon (0.5 g) for 1.5 min. The reaction went cleanly, so more 1,1-dimethylethyl (3R)-3-{[(2-chloro-5-nitrophenyl)sulfonyl]amino}-1-pyrrolidinecarboxylate (5.55 g, 13.67 mmol) in EtOAc (200 ml) was hydrogenated (room temp, 1 atm) over 5% platinum on carbon (3.4 g). After 15 min, the reaction was filtered through celite and washed with EtOAc (50 ml). The solvents were evaporated in vacuo, and the residue was purified by flash chromatography (100 g silica cartridge, 100:0 to 0:100 cyclohexane/EtOAc, 60 min) to afford the title compound (5.46 g). LC-MS: m/z, 376 (M+H), rt 2.91 min.
To 2-methyl-5-nitrobenzenesulfonyl chloride (11.78 g, 50 mmol) in DCM (80 mL) was added pyridine (4.85 ml, 60 mmol) followed by 1,1-dimethylethyl (3R)-3-amino-1-pyrrolidinecarboxylate (9.3 g, 50 mmol). The reaction mixture was stirred at room temperature for 18 hours. The solvent was evaporated to small volume, and that residue purified by flash chromatography (silica cartridge, 100% DCM to 75% DCM/25% EtOAc). The product containing fractions were combined and purified further with flash chromatography (silica cartridge, 97% DCM/3% (2M NH4) MeOH) to afford the title compound (12 g).
A suspension of 1,1-dimethylethyl (3R)-3-{[(2-methyl-5-nitrophenyl)sulfonyl]amino}-1-pyrrolidinecarboxylate (11.55 g, 30 mmol) in EtOH (500 ml) was added to a flask containing 10% Pd/C (1.1 g) and the reaction mixture was hydrogenated at room temperature overnight. The catalyst was filtered off, washed thoroughly with EtOH, THF, and DCM/MeOH (1/1), and the filtrate evaporated. The residue was dissolved in DCM/MeOH (9:1) and passed through a pad of silica. The filtrate was evaporated to afford the title compound (9.1 g).
A mixture of 2-bromo-5-nitrobenzene (5 g, 25 mmol) and chlorosulfonic acid (15 ml) was stirred and heated at 130° C. for 24 h. The reaction mixture was cooled down to room temperature for 12 hours and then poured carefully over ice. Once the ice melted, the mixture was filtered to afford the title compound (7.33 g). This solid was used crude in the next steps. LC-MS: m/z, 300 (M+H), rt 1.45 min.
A solution of 2-bromo-5-nitrobenzenesulfonyl chloride (0.100 g, 0.33 mmol), triethylamine (0.138 ml, 1.0 mmol) and 1,1-dimethylethyl (3R)-3-amino-1-pyrrolidinecarboxylate (0.059 ml, 0.34 mmol) in DCM (2 mL) was stirred at room temperature for 16 hours. The reaction was diluted with DCM, washed with 1 N HCl, water, and sat NaHCO3. The organic layer was dried with MgSO4 and evaporated to afford the title compound (0.058 g). LC-MS: m/z, 450 (M+H), rt 1.54 min.
To a solution of 1,1-dimethylethyl (3R)-3-({[5-amino-2-(methyloxy)phenyl]sulfonyl}amino)-1-pyrrolidinecarboxylate (0.063 g, 0.17 mmol)) in DCM (2 mL) was added ethyl isocyanate (0.017 ml, 0.20 mmol). The reaction mixture was shaken at room temperature overnight. Triethylamine (0.034 ml, 0.4 mmol) was then added to the mixture and it was stirred at room temperature for 3 hours. More ethyl isocyanate (0.034 ml, 0.40 mmol) was added to the mixture and it was shaken at room temperature overnight. Then water (1.0 ml) was added and shaking was continued for another 3 hours. The mixture is then put through a phase separator to dry it. Then 4N HCl in 1,4-dioxane (1.0 ml) was added and the mixture was shaken overnight, and then blown down to dryness. The residue was diluted with DCM (3 ml), and DIEA (0.118 mL, 0.68 mmol) and shaken for 3 hours. BrCN (0.40 mL, 1.2 mmol) was then added and the solution shaken at room temperature overnight. PS-trisamine resin (0.100 g, 0.34 mmol/g) was added to the reaction and stirred overnight. Then the mixture was filtered, concentrated and the concentrate purified by preparatory HPLC (without TFA) to afford the title compound (0.0059 g). LC-MS: m/z, 368 (M+H), rt 1.33 min.
To a solution of 1,1-dimethylethyl (3R)-3-({[5-amino-2-(methyloxy)phenyl]sulfonyl}amino)-1-pyrrolidinecarboxylate (0.063 g, 0.17 mmol)) in DCM (2 mL) was added DIEA (0.118 ml, 0.68 mmol) and then cooled to 0° C. Phenyl isocyanate (0.022 ml, 0.20 mmol) was added to the reaction and it was shaken at room temperature over the weekend. Water (1.0 ml) was added and the flask was shaken for 3 hours. The reaction was then put through a phase separator to remove the water. Then 4N HCl in 1,4-dioxane (1.0 ml) was added and the reaction was shaken overnight and then blown down to dryness. The residue was diluted with DCM (3 ml), and DIEA (0.118 mL, 0.68 mmol) and shaken for 3 hours. BrCN (0.113 mL, 0.34 mmol) was then added and the mixture was shaken at room temperature overnight. PS-trisamine resin (0.095 g, 0.34 mmol/g) was added to the reaction and stirred over the weekend. Then the reaction was filtered, concentrated and the concentrate purified by preparatory HPLC (without TFA) to afford the two title compounds, Example 29 (0.0122 g). LC-MS: m/z, 416 (M+H), rt 1.76 min and Example 30 (0.0113 g). LC-MS: m/z, 297 (M+H), rt 1.22 min.
To 2-bromo-5-(trifluoromethyl)benzenesulfonyl chloride (0.0974 g, 0.30 mmol) was added a solution of 1,1-dimethylethyl (3R)-3-amino-1-pyrrolidinecarboxylate (0.0528 g, 0.27 mmol) in DCM (3 mL) followed by triethylamine (0.150 ml, 1.06 mmol). The reaction was shaken at room temperature overnight. Water was added (2.0 ml) and the reaction was shaken for 3 hours. The reaction was put through a phase separator to remove the water, then concentrated. Then 4N HCl in 1,4-dioxane (1.0 ml) was added. After stirring overnight, the mixture was blown down to dryness, diluted with DCM (2.0 ml), and mixed with DIEA (0.187 mL, 1.08 mmol). After shaking for 1 hour, BrCN (0.180 mL, 0.54 mmol) was added, and the mixture was shaken at room temperature overnight. Then PS-trisamine resin (0.260 g, 2.9 mmol/g) was added, then the reaction was filtered, concentrated and the concentrate purified by preparatory HPLC (without TFA) to afford the title compound (0.0558 g). LC-MS: m/z, 397 (M+H), rt 1.85 min.
Using the procedure described in Example 31, the following examples in Table 1 were synthesized, replacing 2-bromo-5-(trifluoromethyl)benzenesulfonyl chloride with the relevant sulfonyl chloride.
To a vial containing 3-hydroxybenzaldehyde (0.083 g, 0.68 mmol) was added a solution of 1,1-dimethylethyl (3R)-3-({[5-amino-2-(methyloxy)phenyl]sulfonyl}amino)-1-pyrrolidinecarboxylate (0.063 g, 0.17 mmol) in DCM/MeOH (2/1 mL), followed by addition of ZnCl2 (0.0023 g, 0.017 mmol) and MP-CNBH3 (2.5-4.5 mmol/g, 0.200 g, 0.68 mmol). The reaction was shaken overnight at room temperature, then heated overnight at 45° C. Then MP-CNBH3 (2.5-4.5 mmol/g, 0.100 g, 0.34 mmol) was added and shaking was continue overnight. Then PS-trisamine (3.6 mmol/g, 0.190 mg, 0.68 mmol) was added and the mixture was shaken for 3 days at room temperature. 4N HCl in 1,4-dioxane (0.700 ml) was added and, after being shaken overnight, the mixture was blown down to dryness. The residue was taken up in DCM (3.0 ml), and mixed with DIEA (0.120 mL, 0.68 mmol). After shaking for 3 hour, BrCN (0.112 mL, 0.34 mmol) was added, and the mixture was shaken at room temperature overnight. PS-trisamine resin (3.6 mmol/g, 0.094 g, 0.34 mmol) was added to the mixture and it was shaken overnight. Then the reaction was filtered, concentrated and the concentrate purified by MDAP (without TFA) to afford the title compound (0.0069 g). LC-MS: m/z, 403 (M+H), rt 1.542 min.
Using the procedure described in Example 39, the following examples in Table 2 were synthesized, replacing 3-hydroxybenzaldehyde with the relevant aldehyde.
To a vial containing 2-furancarbaldehyde (0.065 g, 0.68 mmol) was added a mixture of 2/1 DCM and MeOH (1 mL) followed by AcOH (0.015 ml, catalytic). Then a solution of 1,1-dimethylethyl (3R)-3-{[(5-amino-2-methylphenyl)sulfonyl]amino}-1-pyrrolidinecarboxylate (0.063 g, 0.17 mmol) in 2DCM/1MeOH (2 mL), followed by MP-CNBH3 (2.5-4.5 mmol/g, 0.200 g, 0.68 mmol) was added. The mixture was shaken over the weekend at room temperature. Then MP-CNBH3 (2.5-4.5 mmol/g, 0.040 g, 0.136 mmol) and DMSO (0.015 ml) were added and mixture was shaken overnight. The resin was filtered off and 4N HCl in 1,4-dioxane (0.700 ml) was added. After shaking the flask overnight, the reaction was blown down to dryness under vacuum. The residue was then taken up in DCM (3.0 ml), and mixed with DIEA (0.120 mL, 0.68 mmol). After shaking for 3 hour, BrCN (0.112 mL, 0.34 mmol) was added, and the mixture was shaken at room temperature overnight. PS-trisamine resin (3.6 mmol/g, 0.094 g, 0.34 mmol) was added and the solution was shaken over the weekend. Then the mixture was filtered, concentrated and the concentrate purified by preparatory HPLC (without TFA) to afford the title compound (0.0289 g). LC-MS: m/z, 360.96 (M+H), rt 1.652 min.
Using the procedure described in Example 55, the following examples in Table 3 were synthesized, replacing 2-furancarbaldehyde with the relevant aldehyde.
To a vial containing 2-furancarbaldehyde (0.065 g, 0.68 mmol) was added to a 2/1 mixture of DCM and MeOH (1 mL) followed by AcOH (0.015 ml, catalytic), then a solution of 1,1-dimethylethyl (3R)-3-{[(5-amino-2-chlorophenyl)sulfonyl]amino}-1-pyrrolidinecarboxylate (0.063 g, 0.17 mmol) in a 2/1 mixture of DCM and MeOH (2 mL), followed by MP-CNBH3 (2.5-4.5 mmol/g, 0.200 g, 0.68 mmol). The mixture was shaken over the weekend at room temperature. Then MP-CNBH3 (2.5-4.5 mmol/g, 0.040 g, 0.136 mmol) and DMSO (0.015 ml) were added and the flask was shaken overnight. The resin was filtered off after which 4N HCl in 1,4-dioxane (0.700 ml) was added. The flask overnight and solvent removed under vacuum. The residue was then taken up in DCM (3.0 ml), and mixed with DIEA (0.120 mL, 0.68 mmol). After shaking for 3 hour, BrCN (0.112 mL, 0.34 mmol) was added, and the reaction was shaken at room temperature overnight. Then PS-trisamine resin (3.6 mmol/g, 0.094 g, 0.34 mmol) was added to the, and shaking continued over the weekend. Then the mixture was filtered, concentrated and the concentrate purified by preparatory HPLC (without TFA) to afford the title compound (0.0094 g). LC-MS: m/z, 380.97 (M+H), rt 1.73 min.
Using the procedure described in Example 68, the following examples in Table 4 were synthesized, replacing 2-furancarbaldehyde with the relevant aldehyde.
To a solution of 1,1-dimethylethyl (3R)-3-({[5-amino-2-(methyloxy)phenyl]sulfonyl}amino)-1-pyrrolidinecarboxylate (0.063 g, 0.17 mmol) in DCM (2 mL) was added DIEA (0.118 ml, 0.68 mmol) followed by the addition of 2-(methyloxy)benzoyl chloride (0.027 ml, 0.020 mmol) after cooling to 0° C. The reaction was shaken over weekend at room temperature. Then H2O (1 ml) was added and the mixture was shaken for 3 hours. The resulting material was passed through a hydrophobic frit. Then 4N HCl in 1,4-dioxane (1.0 ml) was added, shaken overnight and blown down to dryness. The residue was then diluted with DCM (3.0 ml), and mixed with DIEA (0.118 mL, 0.68 mmol). After shaking for 3 hour, BrCN (0.113 mL, 0.34 mmol) was added, and the flask was shaken at room temperature overnight. PS-trisamine resin (3.6 mmol/g, 0.100 g, 0.36 mmol) was added to the flask and it was shaken over the weekend. The resulting material was filtered, concentrated and the concentrate purified by preparatory HPLC (without TFA) to afford the title compound (0.0394 g). LC-MS: m/z, 431 (M+H), rt 1.83 min.
To a solution of 1,1-dimethylethyl (3R)-3-({[5-amino-2-(methyloxy)phenyl]sulfonyl}amino)-1-pyrrolidinecarboxylate (0.063 g, 0.17 mmol) in DCM (3.0 mL) cooled to 0° C., were added DIEA (0.118 ml, 0.68 mmol) and [4-(methyloxy)phenyl]acetyl chloride (0.031 ml, 0.020 mmol). The reaction was stirred for 2 hours, then H2O (1 ml) was added. The reaction was passed through a hydrophobic frit. Then 4N HCl in 1,4-dioxane (1.0 ml) was added. After stirring overnight, the reaction was blown down to dryness. The residue was then taken up in DCM (3.0 ml), and mixed with DIEA (0.118 mL, 0.68 mmol). After stirring for 3 hour, BrCN (0.111 mL, 0.34 mmol) was added, and the mixture was stirred at room temperature overnight. PS-trisamine resin (3.6 mmol/g, 0.098 g, 0.34 mmol) was added to the flask and it was shaken overnight. The resulting material was filtered, concentrated and the concentrate purified by preparatory HPLC (without TFA) to afford the title compound (0.0195 g). LC-MS: m/z, 445 (M+H), rt 1.70 min.
To a vial containing 4-pentenoyl chloride (0.0309 g, 0.026 mmol) was added a solution of 1,1-dimethylethyl (3R)-3-({[5-amino-2-(methyloxy)phenyl]sulfonyl}amino)-1-pyrrolidinecarboxylate (0.063 g, 0.17 mmol) and DIEA (0.118 ml, 0.68 mmol) in DCM (3.0 mL) at 0° C. The reaction was shaken for 5 hours, then H2O (2.0 ml) was added and the reaction was shaken for over weekend. The reaction was passed through a hydrophobic frit. 4N HCl in 1,4-dioxane (0.700 ml) was added. After shaking for 5 hours, the reaction was blown down to dryness. The residue was taken up in DCM (3.0 ml), and mixed with DIEA (0.125 mL, 0.68 mmol). After shaking for 3 hour, BrCN (0.112 mL, 0.34 mmol) was added, and the flask was shaken at room temperature overnight. PS-trisamine resin (3.6 mmol/g, 0.094 g, 0.34 mmol) was added to the flask which was shaken overnight. The resulting material was filtered, concentrated and the concentrate purified by MDAP (without TFA) to afford the title compound (0.0147 g). LC-MS: m/z, 379 (M+H), rt 1.512 min.
Using the procedure described in Example 86, the following examples in Table 5 were synthesized, replacing 4-pentenoyl chloride with the relevant acyl chloride.
To a flask containing 5-chloro-1-methyl-1H-pyrazole-4-carbonyl chloride (0.0478 g, 0.026 mmol) was added a solution of 1,1-dimethylethyl (3R)-3-{[(5-amino-2-methylphenyl)sulfonyl]amino}-1-pyrrolidinecarboxylate (0.060 g, 0.17 mmol) and DIEA (0.68 mmol) in DCM (3.0 mL) at 0° C. The flask was shaken overnight, then PS-trisamine resin (3.6 mmol/g, 0.140 g, 0.52 mmol) was added to the flask, and it was shaken overnight. The resin was filtered off, and H2O (1 ml) was added and the flask shaken overnight. The resulting mixture was passed through a hydrophobic frit and 4N HCl in 1,4-dioxane (0.700 ml) was added to the solution. After shaking overnight, the contents of the flask were blown down to dryness and the residue was taken up in DCM (3.0 ml), and mixed with DIEA (0.120 mL, 0.68 mmol). After shaking for 3 hour, BrCN (0.112 mL, 0.34 mmol) was added, and shaking continute at room temperature overnight. Then PS-trisamine resin (3.6 mmol/g, 0.094 g, 0.34 mmol) was added, and shaking continued overnight. Then the contents of the vessel were filtered, concentrated and the concentrate purified by preparatory HPLC (without TFA) to afford the title compound (0.0196 g). LC-MS: m/z, 423 (M+H), rt 1.458 min.
Using the procedure described in Example 103, the following examples in Table 6 were synthesized, replacing 5-chloro-1-methyl-1H-pyrazole-4-carbonyl chloride with the relevant acyl chloride.
To a flask containing 3-(methyloxy)propanoyl chloride (0.0321 g, 0.026 mmol) was added a solution of 1,1-dimethylethyl (3R)-3-{[(5-amino-2-chlorophenyl)sulfonyl]amino}-1-pyrrolidinecarboxylate (0.060 g, 0.17 mmol) and DIEA (0.68 mmol) in DCM (3.0 mL) at 0° C. The flask was shaken for overnight, and then PS-trisamine resin (3.6 mmol/g, 0.140 g, 0.52 mmol) was added, and the resulting mixture was shaken overnight. The resin was filtered off, H2O (1 ml) was added and the flask shaken overnight. The mixture was passed through a hydrophobic frit and 4N HCl in 1,4-dioxane (0.700 ml) was added to the effluent. After shaking overnight, the flask's contents were blown down to dryness, diluted with DCM (3.0 ml), and mixed with DIEA (0.120 mL, 0.68 mmol). After shaking for 3 hour, BrCN (0.112 mL, 0.34 mmol) was added, and the flask was shaken at room temperature overnight. Then PS-trisamine resin (3.6 mmol/g, 0.094 g, 0.34 mmol) was added to the flask and it was shaken overnight. Then the contents of the flask were filtered, concentrated and the concentrate purified by preparatory HPLC (without TFA) to afford the title compound (0.0212 g). LC-MS: m/z, 387 (M+H), rt 1.247 min.
Using the procedure described in Example 118, the following examples in Table 7 were synthesized, replacing 3-(methyloxy)propanoyl chloride with the relevant acyl chloride.
To a chilled (0° C.) solution of 1,1-dimethylethyl (3R)-3-amino-1-pyrrolidinecarboxylate (0.18 ml, 1.061 mmol) and triethylamine (0.42 ml, 3.01 mmol) in DCM (4 ml) was added 2,5-bis[(2,2,2-trifluoroethyl)oxy]benzenesulfonyl chloride (0.4106 g, 1.102 mmol). The reaction mixture was stirred at room temperature overnight. It was diluted with DCM (100 ml), then washed with 0.1M HCl (2×50 ml), sat NaHCO3 (50 ml), and sat NaCl (50 ml). The organic layer was dried MgSO4, filtered and concentrated to afford the title compound (0.4584 g). LC-MS: m/z, 523 (M+H), rt 1.17 min.
To a chilled (0° C.) solution of 1,1-dimethylethyl (3R)-3-amino-1-pyrrolidinecarboxylate (0.18 ml, 1.061 mmol) and triethylamine (0.42 ml, 3.01 mmol) in DCM (4 ml) was added methyl 3-(chlorosulfonyl)-4-(methyloxy)benzoate (0.309 g, 1.167 mmol). The reaction mixture was stirred at room temperature overnight. The mixture was diluted with DCM (100 ml), then washed with 0.1M HCl (2×50 ml), sat NaHCO3 (50 ml), and sat NaCl (50 ml). The organic layer was dried MgSO4, filtered and the effluent concentrated to afford the title compound (0.3468 g). LC-MS: m/z, 415 (M+H), rt 1.01 min.
1,1-Dimethylethyl(3R)-3-[({2,5-bis[(2,2,2-trifluoroethyl)oxy]phenyl}sulfonyl)amino]-1-pyrrolidinecarboxylate (0.1096 g, 0.210 mmol) was dissolved in DCM (2 ml). Then 4N HCl in 1,4-dioxane (1.0 ml) was added. After 2 hours, the solution was blown down to dryness. The residue was then diluted with DCM (3 ml) and DIEA (0.11 mL, 0.629 mmol). The solution was stirred for 10 min before BrCN (0.105 mL, 0.315 mmol) was added. The resultant mixture was stirred at room temperature for 1 hour. Then PS-trisamine resin (1 equiv) was added and stirred for 2 hours after which the mixture was filtered, the effluent concentrated and the concentrate purified by preparatory HPLC (without TFA) to afford the title compound (0.062 g). LC-MS: m/z, 448 (M+H), rt 1.15 min.
1,1-dimethylethyl(3R)-3-[({2-(methyloxy)-5-[(methyloxy)carbonyl]phenyl}sulfonyl)amino]-1-pyrrolidinecarboxylate (0.1 g, 0.241 mmol) was dissolved in DCM (3 ml). 4N HCl in 1,4-dioxane (1.0 ml) was added. After 2 hours, the solution was blown down to dryness, and the residue taken up in DCM (3 ml) and DIEA (0.13 mL, 0.744 mmol). This solution was stirred for 10 min before BrCN (0.13 mL, 0.39 mmol) was added. The resultant mixture was stirred at room temperature for 2 hour. Then PS-trisamine resin (1 equiv) was added and the mixture stirred for 2 hours. Then the solids were filtered off, the effluent concentrated and the concentrate purified by preparatory HPLC (without TFA) to afford the title compound (0.0492 g). LC-MS: m/z, 340 (M+H), rt 0.77 min.
Using the procedure described in Example 31, the following examples in Table 1 were synthesized, replacing 2-bromo-5-(trifluoromethyl)benzenesulfonyl chloride with the relevant sulfonyl chloride.
The compounds according to formula I are cathepsin C inhibitors, which indirectly inhibit the activity of serine proteases that are activated by cathepsin C, such as NE. The compounds according to formula (IA) or (IB), therefore, are useful in the treatment of COPD and other conditions involving cathepsin C and/or such serine proteases. The biological activity of the compounds according to formula (IA) or (IB) can be determined using any suitable assay for determining the activity of a candidate compound as a cathepsin C inhibitor or for determining the ability of a candidate compound to prevent the cathepsin C mediated activation of certain serine proteases, as well as suitable tissue and in vivo models.
All examples were and found to be cathepsin C inhibitors; the pIC50 values for the instant compounds was between 5 and 9.2.
A. Transpeptidation of Leucine-Leucine-O-Methyl (LLOM) cell-based Luminescence Viability Assay
Cathepsin C has been shown to catalyze the transpeptidation of dipeptidyl methyl-O-esters within the lysosome of cell form the monocytic lineage like HL60, U937 or THP1 causing a membranolythic effects that results in cell death (D L. Thiele P. Lipsky PNAS 1990 Vol. 87, pp. 83-87). This phenomenon was used to assess cellular Cathepsin C activity in the presence of our compounds.
Reagents:
Protocol:
In a 96-welled v-bottom plate (polypropylene): 20 uL stock compound solution (10 mM in DMSO) added to wells in top row. DMSO added to alternating rows at 20 and 23 uL. Dilutions are made by placing 10 uL to the row below, mixing then repeating serially until reaching the bottom row using a multichannel pipettor.
The activity of recombinant human cathepsin C is measured by the cleavage of a fluorogenic substrate, H-Ser-Tyr-AMC. Briefly, 20 pM cathepsin C is incubated with test compound (e.g. inhibitor) in a buffer consisting of 50 mM sodium acetate, 30 mM sodium chloride, 1 mM CHAPS, 1 mM dithiothreitol, 1 mM EDTA, pH 5.5 at room temperature for one hour. After one hour of incubating test compound with cathepsin C, the activity assay is initiated by the addition of an equal volume of 0.010 mM H-Ser-Tyr-AMC in the same buffer. After one hour, the activity assay is stopped by the addition of 1/10 volume of 10 mM 2-Aldrithiol. The reaction product is measured on a fluorescence reader set at an excitation wavelength of 360 nm and emission wavelength of 460 nm and equipped with a 400 nm dichroic mirror.
These compound are believed to be useful in therapy as defined above and to not have unacceptable or untoward effects when used in compliance with a permitted therapeutic regime.
The foregoing examples and assay have been set forth to illustrate the invention, not limit it. What is reserved to the inventors is to be determined by reference to the claims.
| Number | Date | Country | |
|---|---|---|---|
| 61046041 | Apr 2008 | US |
