WO2000012082A1 - Formamide compounds as therapeutic agents - Google Patents

Formamide compounds as therapeutic agents Download PDF

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Publication number
WO2000012082A1
WO2000012082A1 PCT/US1999/018960 US9918960W WO0012082A1 WO 2000012082 A1 WO2000012082 A1 WO 2000012082A1 US 9918960 W US9918960 W US 9918960W WO 0012082 A1 WO0012082 A1 WO 0012082A1
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WIPO (PCT)
Prior art keywords
propyl
ethyl
methyl
butyl
compound
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PCT/US1999/018960
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French (fr)
Inventor
Robert Carl Andrews
Marc Werner Andersen
Jennifer Badiang Stanford
Dulce Garrido Bubacz
Joseph Howing Chan
David John Cowan
Michael David Gaul
Darryl Lynn Mcdougald
David Lee Musso
Michael Howard Rabinowitz
Robert William Wiethe
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Glaxo Group Limited
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Priority to AU55749/99A priority Critical patent/AU5574999A/en
Publication of WO2000012082A1 publication Critical patent/WO2000012082A1/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
    • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
    • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D213/72—Nitrogen atoms
    • C07D213/75—Amino or imino radicals, acylated by carboxylic or carbonic acids, or by sulfur or nitrogen analogues thereof, e.g. carbamates
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D277/00—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings
    • C07D277/02—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings not condensed with other rings
    • C07D277/20—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members
    • C07D277/32—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D277/38—Nitrogen atoms
    • C07D277/44—Acylated amino or imino radicals
    • C07D277/46—Acylated amino or imino radicals by carboxylic acids, or sulfur or nitrogen analogues thereof
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D285/00—Heterocyclic compounds containing rings having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by groups C07D275/00 - C07D283/00
    • C07D285/01—Five-membered rings
    • C07D285/02—Thiadiazoles; Hydrogenated thiadiazoles
    • C07D285/04—Thiadiazoles; Hydrogenated thiadiazoles not condensed with other rings
    • C07D285/12—1,3,4-Thiadiazoles; Hydrogenated 1,3,4-thiadiazoles
    • C07D285/125—1,3,4-Thiadiazoles; Hydrogenated 1,3,4-thiadiazoles with oxygen, sulfur or nitrogen atoms, directly attached to ring carbon atoms, the nitrogen atoms not forming part of a nitro radical
    • C07D285/135—Nitrogen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
    • C07D409/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings
    • C07D409/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
    • C07D417/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings linked by a chain containing hetero atoms as chain links

Definitions

  • the present invention provides novel compounds, novel compositions, methods of their use and methods of their manufacture, such compounds generally pharmacologically useful as agents in those disease states alleviated by the inhibition or antagonism of matrix metallopro teases, metalloproteases, and/or tumor necrosis factor-alpha (TNF), which pathologically involve aberrant extracellular matrix degradation, shedding of cell surface protein ectodomains, and/or TNF synthesis, such disease states including arthritis, tumor metastasis and diabetes.
  • TNF tumor necrosis factor-alpha
  • the aforementioned pharmacologic activities are useful in the treatment of mammals.
  • the compounds of the present invention can be used in the treatment of rheumatoid arthritis, osteoarthritis, inflammatory bowel syndromes, periodontal disease, aberrant angiogenesis, tumor invasion and metastasis, comeal ulceration and the complications of diabetes.
  • rheumatology oncology, dentistry, opththalmology, gastroenterology, cardiology, neurology, nephrology, infectious disease and endocrinology therapy for such agents.
  • the matrix metalloprotease (MMP) family of zinc endoproteases includes fibroblast collagenase (MMP-1, collagenase-1), neutrophil collagenase (MMP-8, collagenase-2), chondrocyte collagenase (MMP-13, collagenase-3), gelatinases A and B (MMP's 2 and 9), and members of the stromelysin family such as stromelysin-1 (MMP-3), stromelysin-3 (MMP-11), and matrilysm (MMP-7). These enzymes accelerate breakdown of connective tissue by catalyzed resorption of the extracellular matrix.
  • inhibitors of one or more of the matrix metalloproteases would have utility m a wide range of disease states such as in abrogating the initiation of tumor metastasis and angiogenesis and in halting the pathogenesis of demyelmating diseases of the nervous system, multiple sclerosis being one example MMP inhibitors would also find utility in diseases involving connective tissue degradation in the joint, as occurs in osteoarthritis and rheumatoid arthritis. MMP's-1 and -3 have been found in elevated levels in the synovial fluid of patients with rheumatoid arthritis and osteoarthritis.
  • Collagenase-3 (MMP-13) is a member of the family of MMP's which preferentially digest collagen. Collagenase-3 is one of the more newly characterized MMP's; biochemical studies on the recombinant protein have demonstrated that it cleaves type II collagen, the predominant matrix component of articular cartilage, more efficiently than either MMP- 1 or MMP -2 and that it is expressed by chondrocytes in osteoarthritic cartilage These data would implicate collagenase-3 as a significant target in rheumatoid arthritis and osteoarthritis for inhibition by MMP inhibitors.
  • inflammatory/autoimmune diseases including but not limited to rheumatoid arthritis, osteoarthritis, Crohn's disease and other inflammatory bowel diseases, periodontal disease, gingivitis, and comeal ulceration
  • cardiovascular diseases including but not limited to atherosclerosis, and restenosis
  • metabolic diseases including but not limited to complications of diabetes, osteoporosis, and other disorders involving resorption of bone
  • neurologic diseases including but not limited to multiple sclerosis and other demyehnation ailments
  • diseases of cancer and malignancy including but not limited to cancers of the oral cavity and pharynx (lip, tongue, mouth, pharynx), esophagus, stomach, small intestine, large intestine, rectum, liver and biliary passages, pancreas, larynx, lung, bone, connective tissue, skin, colon, breast, cervix uteri, corpus endomet ⁇ um, ovary, prostate, testis
  • MMP inhibitors include some structure activity relationships for a series of carboxylalkylamrne inhibitors. These molecules are exemplary for MMP inhibitors in general. They generally embody a functional group capable of tightly binding the zinc cofactor at the enzyme active site, which is contained within a peptidic or pseudopeptide structure. Zinc binding groups among the MMP inhibitor art have included hydroxamic acid, reverse hydroxamic acid, thiol, carboxylate, and phosphmate.
  • Hydroxamate metalloprotease inhibitors disclosed in the art usually have the following general structure (I):
  • W is a zmc-chelatmg acyl derivative group of the formula -C(0)NHOH (which by convention and in this application are referred to as "forward hydroxamates”) or a zmc- chelatmg substituted amine group of the formula -NH(OH)C(0)R (which by convention and in this application are referred to as "reverse hydroxamates”), where R is usually hydrogen or alkyl
  • forward hydroxamates a zmc-chelatmg substituted amine group of the formula -NH(OH)C(0)R
  • MMP activity in conditions characterized by its overproduction would be of benefit, and compounds which inhibit MMP's would act in this manner at a specific target and be useful and of benefit.
  • the present invention fills this need by providing compounds that are potetn, specific, orally active inhibitors of matrix metalloproteases.
  • Tumor necrosis factor- ⁇ (TNF ⁇ ), hereinafter called “TNF”
  • TNF Tumor necrosis factor- ⁇
  • Human TNF is produced primarily by activated monocytes and macrophages. Human TNF is produced as a larger pro- form of 26 kD which is processed to a secreted 17 kD mature form by proteolytic processing of the alamne-76 - valme-77 peptide bond.
  • TNF secretion in vivo inhibit the cell-associated proteolytic processing of the 26 kD pro-TNF to the mature 17 kD form.
  • the proteolytic activity is thought to reside in an mtracellular or cell-associated specific enzyme or family of enzymes, which by convention is called a "TNF convertase", distinct from the matrix metalloproteases but related in that both contain a zmc cation at the active site TNF convertase enzymatic activity can be detected m monocyte membrane fractions, and the enzyme activity can be inhibited by certain matrix metalloprotease - inhibiting compounds.
  • a metalloprotease is thought to mediate the proteolysis of the cell - surface IgE receptor CD23.
  • Certain of the CD23 - derived peptides possess biological activitites mimicking those of cytokmes, including TNF ⁇ .
  • Metalloprotease - like activity is also thought to contribute to the shedding of certain cell surface protein ectodomains such as L-selectm, fibronectin, thyrotropin stimulating hormone receptor, transforming growth factor alpha precursor, low density hpoprotein receptor, beta amyloid precursor protein, mterleukm-6 receptor alpha subunit, Fas ligand, CD40 ligand, epidermal growth factor receptor, macrophage colony stimulating factor, mterleuk ⁇ n-1 receptor type II, CD30, and tumor necrosis factor receptors type I and II.
  • cell surface protein ectodomains such as L-selectm, fibronectin, thyrotropin stimulating hormone receptor, transforming growth factor alpha precursor, low density hpoprotein receptor, beta amyloid precursor protein, mterleukm-6 receptor alpha subunit, Fas ligand, CD40 ligand, epidermal growth factor receptor, macrophage colony stimulating factor, mterleuk ⁇ n-1 receptor type II
  • TNF is known to mediate many biological responses in vivo. Preclimcal and clinical studies in animals and humans with specific TNF neutralizing antibodies, soluble TNF receptor constructs, and TNF detection techniques have implicated TNF as a mediator m numerous pathologies.
  • the compounds of the present invention by virtue of their activity in inhibiting TNF production and/or their activity in preventing cell surface protein ectodomain shedding should show utility in the treatment of diverse pathologies such as; I) inflammatory/autoimmune diseases, including but not limited to rheumatoid arthritis, osteoarthritis, Crohn's disease and other inflammatory bowel diseases and inflammatory gastrointestinal diseases, and systemic lupus erythematosis, n) reperfusion injuries, such as those caused by an initial ischemic event; m) systemic inflammatory response syndromes, including but not limited to sepsis, burn injury, pancreatitis, and adult respiratory distress syndrome; IV) allergic and dermatologic diseases, including but not limited to delayed type hypersensitivity, psoriasis, asthma, eczema, allergic rhinitis, and allergic conjunctivitis; v) cardiovascular diseases, including but not limited to hyperhpidemia, myocardial infarction.
  • I) inflammatory/autoimmune diseases including but not
  • Atherosclerosis chronic obstructive pulmonary disease, and restenosis
  • metabolic diseases including but not limited to osteoporosis, obesity, and diabetes
  • vn neurologic diseases, including but not limited to Alzheimer's disease, Parkinson's disease, multiple sclerosis, aneurism, and stroke
  • vm transplant rejection, including but not limited to organ transplant rejection and graft versus host disease
  • ix diseases of cancer and malignancy, including but not limited to cancers of the oral cavity and pharynx (lip, tongue, mouth, pharynx), esophagus, stomach, small intestine, large intestine, rectum, liver and biliary passages, pancreas, larynx, lung, bone, connective tissue, skm, colon, breast, cervix uteri, corpus endomet ⁇ um, ovary, prostate, testis, bladder, kidney and other urinary tissues, eye, brain and central nervous system, thyroid and other endoc ⁇ ne gland, leuk
  • TNFc TNF-alpha converting enzyme
  • the present invention fulfills this need by providing potent, orally active inhibitors of shedding of cell surface protein ectodomains acting via inhibition of one or more specific enzymes which mediate this proteolytic event.
  • CD23 proteolysis in conditions characterized by an overabundance of CD23 proteolytic fragments would be of benefit, and compounds which inhibit CD23 proteolysis would be useful and of benefit.
  • the present invention fills this need by providing potent inhibitors of CD23 proteolysis acting via inhibition of one or more specific enzymes which mediate this proteolytic event.
  • A is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, or a direct bond;
  • a 2 is C(0)NR 7 , NR 7 C(0), S0 2 NR 7 , NR 7 S0 2 ,NR 7 , S, SO, S0 2 , O, or a direct bond, where R 7 is as defined below;
  • a 3 is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, or a direct bond;
  • a 4 is alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocyclyl, heteroaryl, aryl, NR 8 R 9 ,
  • D is alkylene, alkenylene, alkynylene, NR 10 (O)C, NR ⁇ 0 , S, SO, S0 2 , O, or a direct bond, where R ]0 is as defined below;
  • D 2 is S, SO, S0 2 , O, C(0)NR u , NR supplementC(0), NR,,, or a direct bond, where R n is as defined below;
  • D 3 isalkylene, alkenylene, alkynylene, arylene, heteroarylene, S, SO, S0 2 , O, C(0)NR ⁇ 2 ,
  • D 4 is aryl, aryloxy, heteroaryl, or heteroaryloxy
  • D 5 and D 6 are, independently, lower alkylene, O, S, SO, or S0 2 ;
  • R 3 is hydrogen or lower alkyl;
  • E] is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, C(0)NR ⁇ , or a direct bond, where R ]3 is as defined below;
  • E is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, NR U , S, SO, S0 2 , O, C(O), or a direct bond, where R 14 is as defined below;
  • E 3 is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, NR 15 , S, SO, S0 2 , O, C(O),
  • R )5 and R 16 are as defined below;
  • E 4 is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, NR 18 , S, SO, SO,, O, N(R 18 )C(0), C(0)N(R 18 ), C(O),
  • R ]7 and R !8 are as defined below;
  • E 5 is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, NR, 9 , S, SO, S0 2 , O, C(O),
  • R ]9 and R 20 are as defined below;
  • E 6 is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, NR 2 ⁇ , S, SO, S0 2 , O, C(O), or a direct bond, where R 2! is as defined below;
  • E 7 is hydrogen, NR 22 R 23 , OR 22 , SR 22 , SOR 22 , S0 2 R 22 , alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocyclyl, aryl, or heteroaryl, where R 22 and R 23 are as defined below;
  • R 5 is hydrogen or lower alkyl; Re is where Zi is heteroarylene;
  • Z 2 is lower alkylene, lower alkenylene, lower alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, C(0)NR 24 , NR 24 C(0), S0 2 NR 24 , NR 24 S0 2 , NR 24 , S, SO, S0 2 , O, C(O), C(0)0, OC(O), or a direct bond, where R 24 is as defined below;
  • Z 3 is alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocyclyl, heteroaryl, aryl, NR 25 R 26 , OR 25 , or hydrogen, where R 25 and R 26 are as defined below;
  • R7, Rg, R 9 , Rio, Ri b Rl2; Rl3 > Rl4, Rl5 > Rl6) Rl7 > Rl8, l9, R20, R2I ) ⁇ 2 > R23 > R24, R25) an d R 2 6 are, independently, hydrogen, alkyl, alkynyl, alkenyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, or heteroaryl; or a pharmaceutically acceptable salt, solvate, biohydrolyzable ester, biohydrolyzable amide, affinity reagent, or prodrug thereof.
  • the present invention provides a family of compounds having the general structural formula:
  • W is a reverse hydroxamic acid group
  • Ri is a substituent other than hydrogen; i is a hpophilic substituent preferably with ste ⁇ c bulk proximal to the peptide backbone, and; R 6 is a heteroaryl substituent.
  • Such compounds are novel and are unknown in the art and, given the appropriate choice of Ri, R 2 , R 3 , t , R 5 , and R ⁇ ;, as described herein, show potent inhibition of MMP's, cell-free TNF convertase enzyme and TNF release from cells, and m some cases inhibit TNF convertase and TNF release from cells in preference to matrix metalloproteases.
  • the heteroaryl nature of Pv, in combination with an appropriate choice of R 1 ; R 2 , R 3 , R 4 , and R 5 as described herein is beneficial in achieving increased potency against TNF release from cells relative to inhibition of MMP's Such molecules can be selective for TNF inhibition over MMP's and can possess an improved therapeutic profile where inhibition of one or more of the matrix metalloproteases is associated with an adverse biological response or abnormal pathology.
  • R 3 , R_ t , and R 5 as described herein is also beneficial in achieving selective inhibition of one or more of the matrix metalloproteases (for example, collagenase-3) in preference to TNF convertase inhibition and inhibition of TNF release from whole cells.
  • matrix metalloproteases for example, collagenase-3
  • A) is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, or a direct bond;
  • a 3 is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, or a direct bond;
  • a t is alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocyclyl, heteroaryl, aryl, NR 8 R 9 , OR 8 or hydrogen, where R 8 and R 9 are as defined below;
  • alkylene alkenylene, alkynylene, NR 10 (O)C, NR 10 , S, SO, S0 2 , O, or a direct bond, where Rio is as defined below;
  • D 3 ⁇ s alkylene, alkenylene, alkynylene, cycloalkylene, arylene, heteroarylene, S, SO, S0 2 , O, C(0)NR, 2 , NR, 2 C(0), S0 2 NR 12 , NR, 2 S0 2 , NR, 2 , or a direct bond, where R, 2 is as defined below;
  • D 4 is aryl, aryloxy, heteroaryl, or heteroaryloxy
  • D 5 and D 6 are, independently, lower alkylene, O, S, SO, or S0 2 ;
  • R 3 is hydrogen or lower alkyl
  • Ei is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, C(0)NR ⁇ 3 , or a direct bond, where R ⁇ is as defined below;
  • E 2 is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, NR, 4 , S, SO, S0 2 , O, C(O), or a direct bond, where R is as defined below;
  • E 3 is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, NR 15 , S, SO, S0 2 , O, C(O),
  • R ⁇ and R ⁇ 6 are as defined below;
  • E 4 is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, NR, 8 , S, SO, S0 2 , O, N(R, 8 )C(0), C(0)N(R 18 ), C(O),
  • R and R ]8 are as defined below;
  • E 5 is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, NR 19 , S, SO, S0 2 , O, C(O),
  • R, 9 and R 20 are as defined below;
  • E 6 is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, NR 2] , S, SO, S0 , O, C(O), or a direct bond, where R 2 ⁇ is as defined below;
  • E 7 is hydrogen, NR 22 R 23 , OR 22 , SR 22 , SOR 22 , S0 2 R 22 , alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocyclyl, aryl, or heteroaryl, where R 22 and R 23 are as defined below;
  • R 5 is hydrogen or lower alkyl
  • Z] is heteroarylene
  • Z 3 is alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocyclyl, heteroaryl, aryl, NR 25 R 2 6,
  • Preferred embodiments of the invention include compounds of general formula (II) where
  • Ri is methyl, t ⁇ fluoromethyl, ethyl, isopropyl, n-propyl, tert-butyl, 3- methoxycyclopentyl, furan-2-ethynyl, 4-methyl-l -pentyl, 2-th ⁇ ophenesulfanylmethyl, 4- trifluoromethylcyclohexyl, 3-am ⁇ nophenoxymethyl, 3-(4-mo holme)-l -propyl, 2-(3- tetrazolyl)-l -ethyl, 2-(3-pyr ⁇ dyl)-l -ethyl, 2-(3-furyl)-l -ethyl, 2-(2-th ⁇ azolyl)-l -ethyl, 3,3,3- tr ⁇ fluoro-1 -propyl, 4,4,4-tr ⁇ fluoro- 1 -butyl, 2-(4-tr ⁇ fluorophenyl)-l -
  • R 2 is 5-methylth ⁇ ophene-2-methyl, 2-furanmethyl, th ⁇ ophene-2-methyl, benzoth ⁇ ophene-2-methyl, benzofuran-2-methyl, 4-fluorobenzyl, 3-phenyl-l -propyl, 3- phenyl-2-methyl-l -propyl, 3-(2-pyr ⁇ dyl)-l -propyl, 3-(th ⁇ ophene-2-yl)-l -propyl, 4-phenyl-l- butyl, 3-phenyl-2-propene-l-yl, 3-(benzofuran-3-yl)-l-propyl, 3-(benzoth ⁇ ophene-3-yl)-l- propyl, 3-(furan-2-yl)-l -propyl, 3 -(2-th ⁇ azolyl)-l -propyl, 3-(py ⁇ m ⁇ dm-2-yl)-l -propyl, 3-phenyl-2-ethyl
  • R 3 is hydrogen, isobutyl, or methyl
  • R is tert-butyl, 1-propoxy-l -ethyl, 4-(benzyloxycarbonylam ⁇ no)-l -butyl, 2-(2- (benzyloxycarbonylammo)- 1 -ethylsulfanyl)-2-propyl, 4-(2-pyr ⁇ dylcarbonylammo)-2-methyl- 2-butyl, 4- ⁇ sobutoxycarbonylam ⁇ no-l -butyl, 3-py ⁇ dylmethyl, 3-(2-th ⁇ ophenecarbonylam ⁇ no)- 2-methyl-2-propyl, 4-propoxycarbonylammo-2-butyl, 4-(2-naphthylacetylammo)-l -butyl, 1- ethoxycarbonylammo- 1 -ethyl, 4-(2-pyr ⁇ dylcarbonylam ⁇ no)-2-methyl-2-butyl, 3- (benzyloxycarbonylam ⁇ no)-l
  • Re is 2-th ⁇ azolyl, 2-py ⁇ dyl, 2-py ⁇ m ⁇ d ⁇ nyl, 2-(l,3,4-th ⁇ ad ⁇ azolyl), 3-(5- methyhsoxazolyl), 3-pyr ⁇ dyl, 3-mdolyl, 2-(5-methylth ⁇ azolyl), 3-hydroxy-2-pyr ⁇ dyl, 2-(5- ethanesulfanyl-l,3,4-th ⁇ ad ⁇ azolyl), 2-benzoth ⁇ azolyl, 6-methoxy-2-benzoth ⁇ azolyl, 4-pyr ⁇ dyl, 2-pyrazmyl, 3-qu ⁇ nol ⁇ nyl, 2-(5-methyl-l,3,4-th ⁇ ad ⁇ azolyl), 6-ammo-3-pyr ⁇ dyl, 6-ammo-2- py ⁇ dyl, 4-ammo-3-pyr ⁇ dyl, or 3-ammo-2-pyr ⁇ dyl.
  • Particularly preferred embodiments of the invention include compounds of general formula (II) where R, is methyl, ethyl, n-propyl, isopropyl, 4-methyl- 1 -pentyl, 2- thiophenesulfanylmethyl, 3-am ⁇ nophenoxymethyl, 2-(3-tetrazolyl)-l -ethyl, 2-(3-pyr ⁇ dyl)-l- ethyl, 2-(3-furyl)-l -ethyl, 2-(2-th ⁇ azolyl)-l -ethyl, 3,3,3-tr ⁇ fluoro-l-propyl, 2-(4- trifluorophenyl)- 1 -ethyl, th ⁇ ophene-3-ethynyl, 2-mtrophenoxymethyl, 3-mtrophenoxymethyl, or vinyl; R 2 is 5-methylth ⁇ ophene-2-methyl, 2-furanmethyl, th ⁇ ophene-2-methyl, benzoth ⁇ oph
  • R 3 is hydrogen
  • R 4 is tert-butyl, 4-(benzyloxycarbonylammo)- 1 -butyl, 2-(2- (benzyloxycarbonylamino)- 1 -ethylsulfanyl)-2-propyl, 3-pyr ⁇ dylmethyl, 4-(2- naphthylacetylamino)- 1 -butyl, 3 -(benzyloxycarbonylamino)- 1 -propyl, 3-carbamoylam ⁇ no- 1 - propyl, 4-hydroxybenzyl, 4-chlorobenzyl, 4-fluorobenzyl, 3-( ⁇ m ⁇ no-(2,3,6-t ⁇ methyl-4- methoxybenzenesulfonylammo))-methylam ⁇ no- 1 -propyl, 4-benzyloxycarbonylammobenzyl, isopropyl, cyclohexyl, 4-cyclopentylacetylam ⁇ no-l -butyl
  • R 5 is hydrogen; and Ri is 2-th ⁇ azolyl, 2-pyr ⁇ dyl, 2-pyr ⁇ m ⁇ dmyl, 2-(l,3,4-th ⁇ ad ⁇ azolyl), 3-(5- methyhsoxazolyl), 3-pyr ⁇ dyl, 2-(5-methylth ⁇ azolyl), 3-hydroxy-2-pyr ⁇ dyl, 2-(5- ethanesulfanyl-l,3,4-th ⁇ ad ⁇ azolyl), 4-pyr ⁇ dyl, 2-pyraz ⁇ nyl, 2-(5-methyl-l,3,4-th ⁇ ad ⁇ azolyl), 6- am ⁇ no-3-pyr ⁇ dyl, 6-ammo-2-pyr ⁇ dyl, 4-ammo-3-pyr ⁇ dyl, or 3-ammo-2-py ⁇ dyl.
  • More particularly preferred embodiments of the invention include compounds of general formula (II) where
  • Ri is methyl, n-propyl, isopropyl, ethyl, or 3,3,3-tr ⁇ fluoro-l-propyl;
  • R 2 is 5-methylth ⁇ ophene-2-methyl, 3-phenyl- 1 -propyl, 4-phenyl- 1 -butyl, 2-phenyl- 1 - ethyl, 4-phenylcyclohexylmethyl, 3-(4-chlorophenyl)-l -propyl, or benzyl;
  • R 3 is hydrogen
  • R) is tert-butyl, isopropyl, 2-butyl, 1-methoxy-l -ethyl, 1-hydroxy-l -ethyl, 2- methanesulfanyl-2 -propyl, or 2-hydroxy-2-propyl;
  • R 5 is hydrogen;
  • R 6 is 2-th ⁇ azolyl, 2-py ⁇ dyl, 3-pyr ⁇ dyl, or 4-pyr ⁇ dyl.
  • the compounds of the present invention are inhibitors of matrix metalloproteases, TNF converting enzyme, and TNF activity from whole cells.
  • the compounds of the present invention may also inhibit shedding of pathologically significant cell surface protein ectodomains.
  • the compounds of the present invention may also inhibit CD23 proteolysis
  • the invention described herein is additionally directed to pharmaceutical compositions and methods of inhibiting matrix metalloprotease and/or TNF activity and/or CD23 proteolytic fragment activity in a mammal, which methods comprise administering to a mammal in need of a therapeutically defined amount of a compound of formula (I) or (II), defined above, as a single or polymorphic crystalline form or forms, an amorphous form, a single enantiomer, a racemic mixture, a single stereoisomer, a mixture of stereoisomers, a single diastereoisomer, a mixture of diastereomers, a solvate, a pharmaceutically acceptable salt, a solvate, a prodrug, a biohydrolyzable ester, or a biohydrolyzable amide thereof.
  • the present invention provides a method of inhibiting a matrix metalloprotease, comprising the step of administering to a mammal m need thereof a pharmacologically effective amount of a compound of the present invention.
  • the invention further provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a pharmacologically effective amount of a compound of the present invention sufficient to inhibit a matrix metalloprotease.
  • a matrix metalloprotease- inhibiting amount can be an amount that reduces or inhibits a matrix metalloprotease activity in the subject
  • a compound of the present invention in the preparation of a medicament to inhibit a matrix metalloprotease
  • the present invention further provides a method of inhibiting the lntracellular release of tumor necrosis factor alpha, comprising the step of administering to a mammal in need thereof a pharmacologically effective amount of a compound of the present invention.
  • the invention further provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a pharmacologically effective amount of a compound of the present invention sufficient to inhibit cellular release of mature tumor necrosis factor.
  • An amount sufficient to inhibit cellular release of mature tumor necrosis factor can be an amount that reduces or inhibits cellular release of mature tumor necrosis factor in the subject.
  • a compound of the present invention m the preparation of a medicament to inhibit the cellular release of mature tumor necrosis factor alpha.
  • Also provided is a method of inhibition of shedding of cell surface protein ectodomains comprising the step of administering to a mammal m need thereof a pharmacologically effective amount of a compound of the present invention.
  • the invention further provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a pharmacologically effective amount of a compound of the present invention sufficient to inhibit shedding of cell surface protein ectodomains.
  • An amount sufficient to inhibit shedding of cell surface protein ectodomains can be an amount that reduces or inhibits shedding of one or more cell surface protein ectodomains, such as L-selectin, fibronectin, thyrotropm stimulating hormone receptor, transforming growth factor alpha precursor, low density lipoprotein receptor, beta amyloid precursor protein, mterleukm-6 receptor alpha subunit, Fas ligand, CD40 ligand, epidermal growth factor receptor, macrophage colony stimulating factor, interleukin- 1 receptor type II, CD30, and tumor necrosis factor receptors type I and II, in the subject.
  • cell surface protein ectodomains such as L-selectin, fibronectin, thyrotropm stimulating hormone receptor, transforming growth factor alpha precursor, low density lipoprotein receptor, beta amyloid precursor protein, mterleukm-6 receptor alpha subunit, Fas ligand, CD40 ligand, epidermal growth factor receptor, macrophage colony stimulating
  • a method of inhibiting CD23 proteolysis comprising the step of administering to a mammal in need thereof a pharmacologically effective amount of a compound of the present invention.
  • the invention further provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a pharmacologically effective amount of a compound of the present invention sufficient to inhibit CD23 proteolysis.
  • An amount sufficient to inhibit CD23 proteolysis can be an amount that reduces or inhibits CD23 proteolysis in the subject.
  • a compound of the present invention m the preparation of a medicament to inhibit CD23 proteolysis.
  • composition comprising a pharmaceutically acceptable carrier and a pharmacologically effective amount of a compound of the present invention sufficient to decrease, or inhibit, a malignant growth.
  • a compound of the present invention in the preparation of a medicament to decrease or inhibit a malignant growth.
  • the present invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a pharmacologically effective amount of a compound of the present invention sufficient to treat arthritis.
  • Such an amount can be an amount that relieves, i.e., reduces or eliminates, one or more physiologic characteristic of arthritis.
  • a compound of the present invention in the preparation of a medicament to treat arthritis.
  • a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a pharmacologically effective amount of a compound of the present invention sufficient to treat diabetes.
  • Such an amount can be an amount that reduces or eliminates one or more of the complications associated with diabetes.
  • the present invention contemplates treating any of these diseases/conditions m a subject by administering to the subject the recited pharmaceutical composition.
  • a compound of the present invention m the preparation of a medicament to treat diabetes.
  • the compounds of the present invention can be administered to any mammal in need of inhibition of matrix metalloprotease activity, CD23 proteolysis, shedding of cell surface protein ectodomains and/or TNF activity.
  • mammals can include, for example, horses, cows, sheep, pigs, mice, dogs, cats, primates such as chimpanzees, gorillas, rhesus monkeys, and, most preferably humans.
  • Certain examples of the invention also are orally bioavailable in animals and possess oral activity in animal models of disease.
  • Salts encompassed within the term "pharmaceutically acceptable salts" refer to non-toxic salts of the compounds of this invention which are generally prepared by reacting the free base with a suitable organic or inorganic acid or by reacting the acid with a suitable organic or inorganic base.
  • Representative salts include the following salts: Acetate,
  • Edisylate Estolate, Esylate, Fumarate, Gluceptate, Gluconate, Glutamate, Glycollylarsamlate, Hexylresorcinate, Hydrabamme, Hydrobromide, Hydroclo ⁇ de, Hydroxynaphthoate, Iodide, Isethionate, Lactate, Lactobionate, Laurate, Malate, Maleate, Mandelate, Mesylate, Methylbromide, Methylnitrate, Methylsulfate, Monopotassium Maleate, Mucate, Napsylate, Nitrate, N-mefhylglucamine, Oxalate, Pamoate (Embonate), Palmitate, Pantothenate, Phosphate/diphosphate, Polygalacturonate, Potassium, Sahcylate, Sodium, Stearate, Subacetate, Succmate, Tannate, Tartrate, Teoclate, Tosylate, T ⁇ ethio
  • the present invention also covers the individual enantiomers of the compounds represented by formula above as mixtures with diastereoisomer s thereof which one or more of the three stereocenters are inverted.
  • lower refers to a group having between one and six carbons.
  • alkyl refers to a straight or branched chain hydrocarbon having from one to ten carbon atoms, optionally substituted with substituents selected from the group consisting of lower alkyl, lower alkoxy, lower alkylsulfanyl, lower alkylsulfenyl, lower alkylsulfonyl, oxo, hydroxy, mercapto, amino optionally substituted by alkyl, carboxy, carbamoyl optionally substituted by alkyl, ammosulfonyl optionally substituted by alkyl, nitro, cyano, halogen, or lower perfluoroalkyl, multiple degrees of substitution being allowed
  • substituents selected from the group consisting of lower alkyl, lower alkoxy, lower alkylsulfanyl, lower alkylsulfenyl, lower alkylsulfonyl, oxo, hydroxy, mercapto, amino optionally substituted by alkyl, carboxy, carbamoyl optionally
  • alkylene refers to a straight or branched chain divalent hydrocarbon radical having from one to ten carbon atoms, optionally substituted with substituents selected from the group consisting of lower alkyl, lower alkoxy, lower alkylsulfanyl, lower alkylsulfenyl, lower alkylsulfonyl, oxo, hydroxy, mercapto, amino optionally substituted by alkyl, carboxy, carbamoyl optionally substituted by alkyl, ammosulfonyl optionally substituted by alkyl, nitro, cyano, halogen, or lower perfluoroalkyl, multiple degrees of substitution being allowed.
  • substituents selected from the group consisting of lower alkyl, lower alkoxy, lower alkylsulfanyl, lower alkylsulfenyl, lower alkylsulfonyl, oxo, hydroxy, mercapto, amino optionally substituted by alkyl, carboxy, carbamo
  • alkenyl refers to a hydrocarbon radical having from two to ten carbons and at least one carbon - carbon double bond, optionally substituted with substituents selected from the group consisting of lower alkyl, lower alkoxy, lower alkylsulfanyl, lower alkylsulfenyl, lower alkylsulfonyl, oxo, hydroxy, mercapto, ammo optionally substituted by alkyl, carboxy, carbamoyl optionally substituted by alkyl, ammosulfonyl optionally substituted by alkyl, nitro, cyano, halogen, or lower perfluoroalkyl, multiple degrees of substitution being allowed.
  • alkenylene refers to an straight or branched chain divalent hydrocarbon radical having from two to ten carbon atoms and one or more carbon - carbon double bonds, optionally substituted with substituents selected from the group consisting of lower alkyl, lower alkoxy, lower alkylsulfanyl, lower alkylsulfenyl, lower alkylsulfonyl, oxo, hydroxy, mercapto, amino optionally substituted by alkyl, carboxy, carbamoyl optionally substituted by alkyl, ammosulfonyl optionally substituted by alkyl, nitro, cyano, halogen, or lower perfluoroalkyl, multiple degrees of substitution being allowed.
  • Examples of “alkenylene” as used herein include, but are not limited to, ethene-l,2-d ⁇ yl, propene-l,3-d ⁇ yl, methylene- 1,1-d ⁇ yl, and the like.
  • alkynyl refers to a hydrocarbon radical having from two to ten carbons and at least one carbon - carbon triple bond, optionally substituted with substituents selected from the group consisting of lower alkyl, lower alkoxy, lower alkylsulfanyl, lower alkylsulfenyl, lower alkylsulfonyl, oxo, hydroxy, mercapto, ammo optionally substituted by alkyl, carboxy, carbamoyl optionally substituted by alkyl, ammosulfonyl optionally substituted by alkyl, nitro, cyano, halogen, or lower perfluoroalkyl, multiple degrees of substitution being allowed.
  • alkynylene refers to a straight or branched chain divalent hydrocarbon radical having from two to ten carbon atoms and one or more carbon - carbon triple bonds, optionally substituted with substituents selected from the group consisting of lower alkyl, lower alkoxy, lower alkylsulfanyl, lower alkylsulfenyl, lower alkylsulfonyl, oxo, hydroxy, mercapto, ammo optionally substituted by alkyl, carboxy, carbamoyl optionally substituted by alkyl, ammosulfonyl optionally substituted by alkyl, nitro, cyano, halogen, or lower perfluoroalkyl, multiple degrees of substitution being allowed.
  • Examples of “alkynylene” as used herein include, but are not limited to, ethyne-l,2-d ⁇ yl, propyne-l,3-d ⁇ yl, and the like.
  • cycloalkyl refers to a ahcyclic hydrocarbon group with one or more degrees of unsaturation, having from three to twelve carton atoms, optionally substituted with substituents selected from the group consisting of lower alkyl, lower alkoxy, lower alkylsulfanyl, lower alkylsulfenyl, lower alkylsulfonyl, oxo, hydroxy, mercapto, ammo optionally substituted by alkyl, carboxy, carbamoyl optionally substituted by alkyl, ammosulfonyl optionally substituted by alkyl, nitro, cyano, halogen, or lower perfluoroalkyl, multiple degrees of substitution being allowed.
  • Cycloalkyl includes by way of example cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, or cyclooctyl, and the like.
  • cycloalkylene refers to an non-aromatic ahcyclic divalent hydrocarbon radical having from three to twelve carbon atoms, optionally substituted with substituents selected from the group consisting of lower alkyl, lower alkoxy, lower alkylsulfanyl, lower alkylsulfenyl, lower alkylsulfonyl, oxo, hydroxy, mercapto, ammo optionally substituted by alkyl, carboxy, carbamoyl optionally substituted by alkyl, ammosulfonyl optionally substituted by alkyl, nitro, cyano, halogen, or lower perfluoroalkyl, multiple degrees of substitution being allowed.
  • cycloalkylene examples include, but are not limited to, cyclopropyl- 1,1-d ⁇ yl, cyclopropyl- 1,2-d ⁇ yl, cyclobutyl- 1,2- diyl, cyclopentyl- 1,3-d ⁇ yl, cyclohexyl- 1,4-d ⁇ yl, cycloheptyl- 1,4-d ⁇ yl, or cyclooctyl- 1,5-d ⁇ yl, and the like.
  • cycloalkenyl refers to a substituted ahcyclic hydrocarbon radical having from three to twelve carbon atoms and at least one carbon-carbon double bond in the ring system, optionally substituted with substituents selected from the group consisting of lower alkyl, lower alkoxy, lower alkylsulfanyl, lower alkylsulfenyl, lower alkylsulfonyl, oxo, hydroxy, mercapto, amino optionally substituted by alkyl, carboxy, carbamoyl optionally substituted by alkyl, ammosulfonyl optionally substituted by alkyl, nitro, cyano, halogen, or lower perfluoroalkyl, multiple degrees of substitution being allowed.
  • cycloalkenylene examples include, but are not limited to, 1 - cyclopentene-3-yl, l-cyclohexene-3-yl, l-cycloheptene-4-yl, and the like.
  • cycloalkenylene refers to a substituted ahcyclic divalent hydrocarbon radical having from three to twelve carbon atoms and at least one carbon-carbon double bond in the ring system, optionally substituted with substituents selected from the group consisting of lower alkyl, lower alkoxy, lower alkylsulfanyl, lower alkylsulfenyl, lower alkylsulfonyl, oxo, hydroxy, mercapto, ammo optionally substituted by alkyl, carboxy, carbamoyl optionally substituted by alkyl, ammosulfonyl optionally substituted by alkyl, nitro, cyano, halogen, or lower perfluoroalkyl, multiple degrees of substitution being allowed
  • substituents selected from the group consisting of lower alkyl, lower alkoxy, lower alkylsulfanyl, lower alkylsulfenyl, lower alkylsulfonyl, oxo, hydroxy, mer
  • heterocyclic or the term “heterocyclyl” refers to a three to twelve-membered heterocyclic ring having one or more degrees of unsaturation containing one or more heteroatomic substitutions selected from S, SO, S0 2 , O, or N, optionally substituted with substituents selected from the group consisting of lower alkyl, lower alkoxy, lower alkylsulfanyl, lower alkylsulfenyl, lower alkylsulfonyl, oxo, hydroxy, mercapto, ammo optionally substituted by alkyl, carboxy, carbamoyl optionally substituted by alkyl, ammosulfonyl optionally substituted by alkyl, nitro, cyano, halogen, or lower perfluoroalkyl, multiple degrees of substitution being allowed
  • Such a ring may be optionally fused to one or more of another "heterocyclic" ⁇ ng(s) or cycloalkyl
  • heterocyclylene refers to a three to twelve- membered heterocyclic ring diradical having one or more degrees of unsaturation containing one or more heteroatoms selected from S, SO, S0 2 , O, or N, optionally substituted with substituents selected from the group consisting of lower alkyl, lower alkoxy, lower alkylsulfanyl, lower alkylsulfenyl, lower alkylsulfonyl, oxo, hydroxy, mercapto, ammo optionally substituted by alkyl, carboxy, carbamoyl optionally substituted by alkyl, ammosulfonyl optionally substituted by alkyl, nitro, cyano, halogen, or lower perfluoroalkyl, multiple degrees of substitution being allowed
  • a ring may be optionally fused to one or more benzene ⁇ ngs or to one or more of another "heterocyclic" rings or cycloalkyl
  • aryl refers to a benzene ring or to an optionally substituted benzene ring system fused to one or more optionally substituted benzene ⁇ ngs, optionally substituted with substituents selected from the group consisting of lower alkyl, lower alkoxy, lower alkylsulfanyl, lower alkylsulfenyl, lower alkylsulfonyl, oxo, hydroxy, mercapto, amino optionally substituted by alkyl, carboxy, tetrazolyl, carbamoyl optionally substituted by alkyl, ammosulfonyl optionally substituted by alkyl, acyl, aroyl, heteroaroyl, acyloxy, aroyloxy, heteroaroyloxy, alkoxycarbonyl, nitro, cyano, halogen, or lower perfluoroalkyl, multiple degrees of substitution being allowed.
  • substituents selected from the group consisting of lower alkyl, lower alk
  • arylene refers to a benzene ring diradical or to a benzene ⁇ ng system diradical fused to one or more optionally substituted benzene ⁇ ngs, optionally substituted with substituents selected from the group consisting of lower alkyl, lower alkoxy, lower alkylsulfanyl, lower alkylsulfenyl, lower alkylsulfonyl, oxo, hydroxy, mercapto, ammo optionally substituted by alkyl, carboxy, tetrazolyl, carbamoyl optionally substituted by alkyl, ammosulfonyl optionally substituted by alkyl, acyl, aroyl, heteroaroyl, acyloxy, aroyloxy, heteroaroyloxy, alkoxycarbonyl, nitro, cyano, halogen, or lower perfluoroalkyl, multiple degrees of substitution being allowed.
  • substituents selected from the group consisting of lower al
  • heteroaryl refers to a five - to seven - membered aromatic ring, or to a polycyclic heterocyclic aromatic ⁇ ng, containing one or more nitrogen, oxygen, or sulfur heteroatoms, where N-oxides and sulfur monoxides and sulfur dioxides are permissible heteroaromatic substitutions, optionally substituted with substituents selected from the group consisting of lower alkyl, lower alkoxy, lower alkylsulfanyl, lower alkylsulfenyl, lower alkylsulfonyl, oxo, hydroxy, mercapto, amino optionally substituted by alkyl, carboxy, tetrazolyl, carbamoyl optionally substituted by alkyl, ammosulfonyl optionally substituted by alkyl, acyl, aroyl, heteroaroyl, acyloxy, aroyloxy, heteroaroyloxy, alkoxycarbonyl, nitro, cyano
  • heteroarylene refers to a five - to seven - membered aromatic ring diradical, or to a polycyclic heterocyclic aromatic ring diradical, containing one or more nitrogen, oxygen, or sulfur heteroatoms, where N-oxides and sulfur monoxides and sulfur dioxides are permissible heteroaromatic substitutions, optionally substituted with substituents selected from the group consisting of lower alkyl, lower alkoxy, lower alkylsulfanyl, lower alkylsulfenyl, lower alkylsulfonyl, oxo, hydroxy, mercapto, ammo optionally substituted by alkyl, carboxy, tetrazolyl, carbamoyl optionally substituted by alkyl, ammosulfonyl optionally substituted by alkyl, acyl, aroyl, heteroaroyl, acyloxy, aroyloxy, heteroaroyloxy, alkoxycarbonyl
  • alkoxy refers to the group R a O-, where R a is alkyl.
  • alkenyloxy refers to the group R a O-, where R a is alkenyl.
  • alkynyloxy refers to the group R a O-, where R a is alkynyl.
  • aryloxy refers to the group R a O-, where R a is aryl.
  • heteroaryloxy refers to the group R a O-, where R a is heteroaryl.
  • alkylsulfanyl refers to the group R a S-, where R a is alkyl.
  • alkenylsulfanyl refers to the group R a S-, where R a is alkenyl.
  • alkynylsulfanyl refers to the group R a S-, where R a is alkynyl.
  • alkylsulfenyl refers to the group R a S(0)-, where R a is alkyl.
  • alkenylsulfenyl refers to the group R a S(0)-, where R a is alkenyl.
  • alkynylsulfenyl refers to the group R a S(0)-, where R a is alkynyl.
  • alkylsulfonyl refers to the group R a S0 2 -, where R a is alkyl.
  • alkenylsulfonyl refers to the group R a S0 -, where R a is alkenyl.
  • alkynylsulfonyl refers to the group R a S0 2 -, where R a is alkynyl.
  • acyl refers to the group R a C(0)- , where R a is alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, or heterocyclyl.
  • aroyl refers to the group R a C(0)- , where R a is aryl.
  • heteroaroyl refers to the group R a C(0)- , where R a is heteroaryl.
  • alkoxycarbonyl refers to the group R a OC(0)-, where R a is alkyl.
  • acyloxy refers to the group R a C(0)0- , where R a is alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, or heterocyclyl.
  • aroyloxy refers to the group R a C(0)0- , where R a is aryl.
  • heteroaroyloxy refers to the group R a C(0)0- , where R a is heteroaryl.
  • the term "optionally” means that the subsequently described event(s) may or may not occur, and includes both event(s) which occur and events that do not occur.
  • substituted refers to substitution with the named substituent or substituents, multiple degrees of substitution being allowed unless otherwise stated.
  • the terms "contain” or “containing” can refer to in-line substitutions at any position along the above-defined alkyl, alkenyl, alkynyl or cycloalkyl substituents with one or more of any of O, S, SO, S0 2 , N, or N-alkyl, including, for example, -CH 2 -0-CH 2 -, -CH 2 -S0 2 -CH 2 -, -CH 2 -NH-CH 3 and so forth.
  • solvate is a complex of variable stoichiometry formed by a solute ( this invention, a compound of formula (I) or (II)) and a solvent.
  • solvents for the purpose of the invention may not interfere with the biological activity of the solute.
  • Solvents may be, by way of example, water, ethanol, or acetic acid.
  • biohydrolyzable ester is an ester of a drug substance (in this invention, a compound of general formula (I) or (II) which either a) does not interfere with the biological activity of the parent substance but confers on that substance advantageous properties in vivo such as duration of action, onset of action, and the like, or b) is biologically inactive but is readily converted in vivo by the subject to the biologically active principle.
  • the biohydrolyzable ester is orally absorbed from the gut and is transformed to (I) or (II) in plasma.
  • biohydrolyzable amide is an amide of a drug substance (in this invention, a compound of general formula (I) or (II) which either a) does not interfere with the biological activity of the parent substance but confers on that substance advantageous properties m vivo such as duration of action, onset of action, and the like, or b) is biologically inactive but is readily converted in vivo by the subject to the biologically active principle.
  • the advantage is that, for example, the biohydrolyzable amide is orally absorbed from the gut and is transformed to (I) or (II) in plasma.
  • Many examples of such are known m the art and include by way of example lower alkyl amides, ⁇ -amino acid amides, alkoxyacyl amides, and alkylaminoalkylcarbonyl amides
  • prodrug includes biohydrolyzable amides and biohydrolyzable esters and also encompasses a) compounds in which the biohydrolyzable functionality in such a prodrug is encompassed in the compound of formula (I) or (II): for example, the lactam formed by a carboxylic group in R 2 and an amme in R 4 , and b) compounds which may be oxidized or reduced biologically at a given functional group to yield drug substances of formula (I) or (II) Examples of these functional groups are, but are not limited to, 1 ,4-d ⁇ hydropy ⁇ d ⁇ ne, N-alkylcarbonyl-l,4-d ⁇ hydropy ⁇ dme, 1,4- cyclohexadiene, tert-butyl, and the like.
  • affinity reagent is a group attached to the compound of formula (I) or (II) which does not affect its in vitro biological activity, allowing the compound to bind to a target, yet such a group binds strongly to a third component allowing a) characterization of the target as to localization within a cell or other organism component, perhaps by visualization by fluorescence or radiography, or b) facile separation of the target from an unknown mixture of targets, whether proteinaceous or not protemaceous
  • An example of an affinity reagent according to b) would be biotin either directly attached to (I) or (II) or linked with a spacer of one to 50 atoms selected from the group consisting of C, H, O, N, S, or P in any combination.
  • An example of an affinity reagent according to a) above would be fluorescein, either directly attached to (I) or (II) or linked with a spacer of one to 50 atoms selected from the group consisting of C, H, O, N, S, or P m any combination
  • pharmacologically effective amount shall mean 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 by a researcher or clinician. This amount can be a therapeutically effective amount Whenever the terms "alkyl” or “aryl” or either of their prefix roots appear in a name of a substituent (e.g. arylalkoxyaryloxy) they shall be interpreted as including those limitations given above for "alkyl” and "aryl”. Alkyl or cycloalkyl substituents shall be recognized as being functionally equivalent to those having one or more degrees of unsaturation.
  • Designated numbers of carbon atoms shall refer independently to the number of carbon atoms in an alkyl, alkenyl or alkynyl or cyclic alkyl moiety or to the alkyl portion of a larger substituent in which the term "alkyl" appears as its prefix root
  • halogen or halo shall include iodine, bromine, chlorine and fluorine.
  • mercapto shall refer to the substituent -SH.
  • cyano shall refer to the substituent -CN.
  • sulfenyl shall refer to the substituent -S(O)-
  • sulfonyl shall refer to the substituent -S(0) 2 -
  • the most preferred compounds of the invention are any or all of those specifically set forth m these examples. These compounds are not, however, to be construed as forming the only genus that is considered as the invention, and any combination of the compounds or their moieties may itself form a genus.
  • the following examples further illustrate details for the preparation of the compounds of the present invention. Those skilled in the art will readily understand that known variations of the conditions and processes of the following preparative procedures can be used to prepare these compounds. All temperatures are degrees Celsius unless noted otherwise.
  • APCI atmospheric pressure chemical lonization
  • ESI electrospray lonization
  • m/z mass to charge ratio
  • MeOH methanol
  • EtOAc ethyl acetate
  • TEA t ⁇ ethylamine
  • DIEA dnsopropylethylamine
  • BOP ( 1 -benzot ⁇ azolyloxy)t ⁇ s(d ⁇ methylam ⁇ no)phosphon ⁇ um hexafluorophosphate
  • LAH lithium aluminum hyd ⁇ de
  • TFA t ⁇ fluoroacetic acid
  • NMM N-methylmorphohne, 4-mefhylmorpholme
  • HMPA hexamethylphospho ⁇ c t ⁇ amide
  • DMPU 1 ,3-d ⁇ methypropylene urea
  • ppm parts per million
  • kD kiloDalton
  • LPS lipopolysaccha ⁇ de
  • EDTA efhylenediamine tetraacetic acid
  • FBS fetal bovme serum
  • PBS phosphate buffered saline solution
  • ELISA enzyme - linked immunosorbent assay
  • Vll " Rj, R 2 , R 3 , R,, R 5 , and R are as defined as for formula (II).
  • RPG is selected from the group consisting of benzyl or 2-tetrahydropyranyl.
  • R 27 is chosen from the group consisting of hydroxyl, 0-C 6 F 5 , or halogen.
  • the conversion of (V) and (VI) to (VII) involves methods known in peptide chemistry; for example, the reaction may be conducted using HOBt in combination with a dehydrating agent such as dicyclohexylcarbodiimide in a suitable solvent, such as DMF.
  • a suitable solvent such as dichloromethane
  • pentafluorophenyl t ⁇ fluoroacetate in the presence of py ⁇ dine
  • EDC and pentafluorophenol in a suitable solvent such as dichloromethane.
  • the displacement reaction to produce (VII) employing (VI) is carried out in the presence of a suitable solvent such as dioxane, THF, dichloromethane, or DMF, at a temperature of 0 °C to 140 °C.
  • a suitable solvent such as dioxane, THF, dichloromethane, or DMF
  • the reaction is effected in the presence of an organic base such as NMM or TEA.
  • R] in (VII) contains a terminal alkynyl group
  • addition of an aryl or heteroaryl bromide to (VII) in the presence of tetrak ⁇ s(t ⁇ phenylphosphme)pallad ⁇ um, Cul, and TEA in DMF or THF affords the alkynylaryl or alkynylheteroaryl intermediate.
  • These Ri substituents may be reduced with palladium on carbon and hydrogen to the saturated species.
  • RPG group where RPG] is benzyl
  • a suitable solvent such as ethanol or THF
  • RPG 2-tetrahydropyranyl
  • Reaction Scheme 2 depicts the synthesis of a compound of formula (IV).
  • R, and R 2 are as defined for formula (II).
  • R 28 0 is a nucleofugal group such as methanesulfonate or p-toluenesulfonate.
  • RPG is defined as for reaction scheme 1.
  • the acid of formula (VIII) may be converted to the alcohol of formula (IX) by treatment with HOBt, O-benzylhydroxylamme hydrochlo ⁇ de or 2- tetrahydropyranyloxyamine, NMM, and a carbodnmide reagent such as EDC in a suitable solvent such as DMF.
  • the alcohol of formula (IX) may be converted to (X) by treatment with methanesulfonyl chloride and py ⁇ dme in a suitable solvent such as dichloromethane.
  • the conversion of (X) to (XI) may be conducted by treatment with potassium carbonate in a suitable solvent such as acetone or 2-butanone, at temperature of 20 °C to 90 °C.
  • (IX) may be converted directly to (XI) by treatment with t ⁇ phenylphosphine and diethyl azodicarboxylate or another azodicarbonyl diester or diamide in a suitable solvent such as THF at a temperature of -78 °C to 50 °C.
  • (XI) where R, contains a primary or secondary hydroxyl group may be treated with a thiol, phenol, or heteroaryl species containing an NH group m the presence of t ⁇ phenylphosphine and diethyl azodicarboxylate or another azodicarbonyl diester or diamide in a suitable solvent such as THF to afford the respective product of hydroxy displacement in (XI).
  • the compound of formula (XI) may be converted to (XII) by treatment with an inorganic base such as sodium hydroxide in water or water in combination with a water - soluble organic cosolvent such as methanol or THF, followed by acidification with an acidic solution such as aqueous citric acid or aqueous sodium bisulfate.
  • the compound of formula (XII) may be converted to (IV) by treatment with acetic anhydride and formic acid or by treatment with formic acetic anhydride in py ⁇ dme in the presence or absence of a suitable cosolvent such as dichloromethane.
  • An alternative route of preparation of compounds of formula (IX) is depicted in reaction Scheme 3.
  • R] and R 2 are as defined as for formula (II).
  • RPG is defined as for reaction scheme 1.
  • R 29 is chosen from the group consisting of lower alkoxy or oxazol ⁇ d ⁇ non-1-yl, where the 4 and 5 positions of an oxazol ⁇ d ⁇ non-1-yl group may be substituted with a lower alkyl, aryl, or lower alkylaryl group and where such an oxazohdinon- 1 -yl substituent may exist as a single stereoisomer or as a mixture of stereoisomers.
  • a carbonyl compound of formula (XIII), where R 29 is an alkoxy group such as methoxy or tert-butoxy, may be treated with a strong base such as LDA m a solvent such as THF at a temperature of from -78 °C to 0 °C, followed by treatment with the aldehyde (XIV) to provide (XV).
  • a strong base such as LDA m a solvent such as THF
  • R 29 is a oxazohdinon- 1-yl substituent
  • treatment of (XIII) with a Lewis acid such as d ⁇ (n-butyl)boron t ⁇ fluoromethanesulfonate in the presence of DIEA in a suitable solvent such as dichloromethane at a temperature of 0 °C followed by addition of the aldehyde (XIV) provides (XV)
  • Treatment of (XV) with aqueous base in the presence or absence of hydrogen peroxide affords (VIII) upon acidification.
  • the acid (VIII) may be converted directly to (IX) as in reaction Scheme 2, or may be treated with a dehydrating agent such a p-toluenesulfonyl chloride m py ⁇ dine or with t ⁇ phenylphosphine and diethyl azodicarboxylate m a suitable solvent such as THF, to afford the lactone (XVI).
  • a dehydrating agent such as a p-toluenesulfonyl chloride m py ⁇ dine or with t ⁇ phenylphosphine and diethyl azodicarboxylate m a suitable solvent such as THF
  • a suitable solvent such as THF
  • Ri and R 2 are as defined as for formula (II).
  • R 30 , R 3 ! , R 32 , and R 33 may be, independently, alkyl, alkenyl, alkynyl, or hydrogen.
  • R 34 and R 35 are selected from the group consisting of aryl, heteroaryl, alkynyl, or alkenyl, with the proviso that the unsaturated carbons of alkenyl and alkynyl groups are directly bonded to Lj
  • L] is selected from the group consisting of bromide, iodide, or t ⁇ fluoromethanesulfonate.
  • L 2 is t ⁇ (lower alkyl)stannyl or -B(OH) 2 .
  • RPG] is defined as for reaction scheme 1.
  • the lactam of general formula (XVII) may be treated with a palladium catalyst such as tetrak ⁇ s(t ⁇ phenylphosph ⁇ ne)pallad ⁇ um and R 34 -L] m a solvent such as acetonit ⁇ le in the presence of a tertiary amine base such as NMM at a temperature of from 20 °C to 200 °C to afford (XVIII).
  • a palladium catalyst such as tetrak ⁇ s(t ⁇ phenylphosph ⁇ ne)pallad ⁇ um and R 34 -L] m a solvent such as acetonit ⁇ le
  • a tertiary amine base such as NMM
  • the olefin in compounds of general formula (XVIII) may be left in place and manipulation of the lactam (XVIII) is carried out as described in reaction Scheme 2 to afford (XIX).
  • (XIX) may be converted to (IV) as desc ⁇ bed in reaction Scheme 2 with or without reduction of the olefin in (XIX), as approp ⁇ ate.
  • the alkyne (XX) may be treated with t ⁇ (butyltm) hydride in the presence of a radical initiator such as azob ⁇ s( ⁇ sobutyron ⁇ t ⁇ le) to afford an alkyl tin intermediate (XXI), which may be freated with R 34 -L] and a catalyst such as Pd(PPh 3 ) 4 m a solvent such as DMF in the presence or LiCl to afford (XXII).
  • XXI may be treated with iodine in an organic solvent such as ether to afford the destannylated vmyl iodide.
  • the iodide may be treated with R 34 -L 2 in the presence of a palladium catalyst such as Pd(PPh 3 ) 4 in a solvent such as DMF or THF at a temperature of 25 °C to 140 °C to afford (XXII).
  • a palladium catalyst such as Pd(PPh 3 ) 4
  • a solvent such as DMF or THF
  • the alkyne (XXIII ) may be treated with R35-L1, CuCl or Cul and an alkylamme such as TEA in the presence of a catalyst such as Pd(PPh 3 ) 4 to afford (XXIV).
  • (XXIV and (XXII) may be manipulated to the intermediate (IV) via the operations described in reaction scheme 2
  • Reaction Scheme 5 depicts the preparation of compounds of general formula (VIII) Fraction Scherre 5
  • Ri and R 2 are as defined for formula (II).
  • R 36 is lower alkyl.
  • RPGi is as defined for reaction Scheme 1.
  • L 3 is bromide, iodide, or t ⁇ fluoromethanesulfonyloxy
  • R 30 , R ⁇ , and R 32 are as defined for reaction scheme 4
  • R 37 and R 38 may be, independently, alkyl, alkenyl, alkynyl, or hydrogen.
  • the ketoester of general formula (XXVII), if not commercially available, may be prepared by reaction of ester (XXV) with a strong base such as LDA followed by treatment with the aldehyde (XIV).
  • the resulting hydroxyester (XXVI) may be used directly or converted to the ketoester (XXVII) by oxidation with, for example, py ⁇ dmium dichromate in a solvent such as dichloromethane
  • the acid chloride (XXVIII) may be condensed with 2,2-d ⁇ methyl-4,6-d ⁇ oxo-l,3-d ⁇ oxane in the presence of py ⁇ dine to afford (XXIX), which may be freated with excess R 35 -OH at a temterature of from 25 °C to 150 °C to provide
  • the ketoester of general formula (XXVII) may be reduced with a reducing agent such as sodium borohyd ⁇ de to afford the hydroxyester (XXVI).
  • a chiral catalyst or chiral ligand in the presence of a reducing agent such as hydrogen or a metal hydride such as borane or LAH may be employed to afford (XXVI) with chiral induction at the newly formed asymmetric center.
  • the alcohol (XXVI) may be converted to (XXX) by treatment with a strong base such as LDA in a suitable solvent such as THF, followed by the addition of R 2 -L 3 in the presence or absence of a cosolvent such as DMPU.
  • (XXX) may be reduced under appropriate conditions. Removal of the ester group by hydrolysis with aqueous hydroxide or, in the case where R 36 is tert-butyl, by treatment with a strong acid such as TFA, affords (VIII) Hydroxy acid (XXXI) is obtained by hydrolysis of the ester group of (XXVI) with aqueous alkali.
  • (XXXI) may be obtained by freatment of (XXVI) with TFA, where R 36 is tert-butyl Coupling of the hydroxy acid (XXXI) with an allylic alcohol (XXXII) m the presence of a dehydrating agent such as EDC and a catalyst such as 4-d ⁇ methylammopy ⁇ dme provides the ester (XXXIII).
  • a dehydrating agent such as EDC
  • a catalyst such as 4-d ⁇ methylammopy ⁇ dme
  • protection of the alcohol functionality of ester (XXVI) with, for example, a tert- butyldimefhylsilyl group may be required before processing of (XXVI) to the acid.
  • R 3 , R4, R 5 , and R 6 are as defined for general formula (II).
  • RPG 2 is selected from the group consisting of tert-butoxycarbonyl, allyloxycarbonyl, or benzyloxycarbonyl .
  • R 39 is selected from the group consisting of 1 -benzot ⁇ azolyloxy, or bromine.
  • the acid of formula (XXXV) may be converted in situ to (XXXVI), where R 39 is bromine, by treatment with bromo-t ⁇ s(pyrrol ⁇ d ⁇ no)phosphon ⁇ um hexafluorophosphate m a suitable solvent such as DMF in the presence of an organic base such as DIEA.
  • the acid of formula (XXXV) may be converted in situ to (XXXVI), where R 39 is benzot ⁇ azolyloxy, by treatment with BOP in a suitable solvent such as DMF in the presence of an organic base such as NMM.
  • the intermediate of formula (XXXV) may be treated with carbonyld ⁇ midazole in a solvent such as dichloromethane, followed by freatment with the amme (XXXVII) to afford (XXXVIII)
  • the intermediate of formula (XXXV) may be treated with HOBt, the amme (XXXVII), an organic base such as NMM, and a carbod ⁇ mide reagent such as EDC in a suitable solvent such as DMF, at a temperature of 0 °C to 80 °C to provide (XXXVIII)
  • the compound of formula (XXXVIII) may be converted to (VI) by deprotection, conditions being particular to the nature of RPG 2 .
  • conversion of (XXXVIII) to (VI) may be accomplished by freatment of (XXXVIII) with TFA in the presence or absence of a suitable solvent such as dichloromethane, at a temperature of 0 °C to 50 °C.
  • Reaction scheme 7 depicts an alternate preparation of an intermediate of general formula (XXXV).
  • R 3 and R are as defined for general formula (II).
  • RPG 2 is as defined for reaction scheme 6.
  • R 40 is lower alkyl or hydrogen.
  • R 41 is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, heterocyclylene, arylene, or heteroarylene, where alkylene, alkenylene, alkynylene, cycloalkylene, and cycloalkenylene substituents may contain one or more O, S, SO, or S0 2 substituents.
  • R 2 is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, heterocyclylene, arylene, O, NH, N-alkyl, or heteroarylene, where alkylene, alkenylene, alkynylene, cycloalkylene, and cycloalkenylene substituents may contain one or more O, S, SO, or S0 2 substituents.
  • R 3 is alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocyclyl, aryl, heteroaryl, or hydrogen, where alkyl, alkenyl, alkynyl, cycloalkyl, and cycloalkenyl substituents may contain one or more O, S, SO, or S0 2 substituents.
  • R» 4 is alkylene or heteroarylene.
  • R 4 5 is alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocyclyl, aryl, heteroaryl, or hydrogen, where alkyl, alkenyl, alkynyl, cycloalkyl, and cycloalkenyl substituents may contain one or more O, S, SO, or S0 2 substituents.
  • the hydroxy compound (XXXIX) may be treated with the reagent R 43 -R 42 -COCI in a solvent such as dichloromethane in the presence of tertiary base such as TEA to afford (XL).
  • (XXXIX) may be freated with R ⁇ -R ⁇ -COOH (where R, 2 is not O, NH, or N- alkyl) and a dehydrating agent such as EDC and a catalyst such as DMAP in a solvent such as DMF or dichloromethane to afford (XL).
  • the compound (XL) where R 4 7 IS NH may be prepared by freating (XXXIX) with R 43 -NCO in a solvent such as dichloromethane.
  • the ether (XLI) may be prepared by treating (XXXLX) with R ⁇ R-nBr or R 45 R 44 I in the presence of a base such as potassium carbonate or sodium hydride in a solvent such as DMF. Removal of the alkyl group R 40 by saponification with aqueous base (or, if appropriate and where R t o is tert-butyl, by treatment with t ⁇ fluoroacetic acid) provides the acid (XXXV).
  • Reaction scheme 8 depicts an alternate preparation of an intermediate of general formula (XXXV).
  • R 3 and R 4 are as defined for general formula (II).
  • RPG 2 is as defined for reaction scheme 6
  • R40, R 4 ⁇ , ⁇ 4 and R45 are as defined for reaction scheme 7.
  • n is 1 to 2.
  • the thioether (XLIII) may be prepared by treating (XLII) with t sR ⁇ Br or ⁇ R ⁇ I and a base such as potassium carbonate or sodium hydride m a solvent such as DMF.
  • the sulfur atom may be oxidized with a reagent such as m-chloroperoxybenzoic acid.
  • Use of one molar equivalent of oxidant may be employed to provide (XLIV) where n is i .
  • Use of two molar equivalents of oxidant may be employed to provide (XLIV) where n is 2.
  • R 3 and R 4 are as defined for general formula (II).
  • RPG 2 is as defined for reaction scheme 6.
  • Rto and R 41 are as defined for reaction scheme 7.
  • R t ⁇ and R 47 are, independently, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocyclyl, aryl, heteroaryl, carboxamidine, or hydrogen, where alkyl, alkenyl, alkynyl, cycloalkyl, and cycloalkenyl substituents may contain one or more O, S, SO, or S0 substituents.
  • R 46 and R 47 may be taken together to constitute a three- to ten-membered ⁇ ng.
  • the thiol (XLII) may be oxidized to the disulfide (XLV) by freatment with a mild base such as TEA and oxygen or air. Either the thiol (XLII) or the disulfide (XLV) may be converted to the sulfonyl chloride (XLVI) by treatment with chlorine gas in tetrachloromethane. Treatment of the sulfonyl chloride (XLVI) with an amme R 46 R 4 7NH in the presence of a tertiary amme base such as TEA or NMM affords (XL VII). Removal of the alkyl group R 40 in (XL VII) by sapomfication with aqueous base (or, if approp ⁇ ate and where
  • R 4 o is tert-butyl, by treatment with frifluoroacetic acid) provides the acid (XXXV)
  • Reaction scheme 10 depicts an alternate preparation of an intermediate of general formula (XXXV).
  • R 3 and R are as defined for general formula (II).
  • RPG 2 is as defined for reaction scheme 6.
  • R 40 and R 41 are as defined for reaction scheme 7.
  • t 6 and R 4 7 are as defined for reaction scheme 9.
  • R ⁇ and R 47 may be taken together to constitute a three- to ten-membered ⁇ ng.
  • the acid (XLVIII) may be converted to the amide (XLIX) by freatment of (XLVIII) and the a ine l ⁇ ⁇ NH with a dehydrating agent such as EDC or BOP in the presence of
  • Reaction Scheme 11 depicts an alternate preparation of an intermediate of general formula
  • R 3 and R 4 are as defined for general formula (II).
  • RPG 2 is defined as for reaction scheme 6.
  • R48 is hydrogen, lower alkyl, or aryl.
  • R, 9 , R 50 , and R 5 are, independently, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocyclyl, aryl, or heteroaryl.
  • R 50 and R 5 ] may be taken together to constitute a five- to ten-membered ⁇ ng.
  • R 49 and R 51 may be taken together to constitute a five- to ten-membered ring.
  • R 48 and R 50 may be taken together to constitute a five- to ten-membered ring
  • R 52 is lower alkyl or benzyl.
  • R 53 is O or NH.
  • L 4 is Br or I.
  • the acid (L) is treated with frifluoroacetic anhydride and TFA to afford the t ⁇ fluoroacetamide, which is then treated with the bromide (LI) and a base such as potassium carbonate or DBU in a solvent such as DMF to provide the ester (LII).
  • (LU) is treated with LDA and aluminum friisopropoxide in the presence of quinidme or quinine in a solvent such as THF at a temperature of from -78 °C to 25 °C to afford (LIII) with a high degree of asymmetric induction.
  • (LV) may be freated with ozone in dichloromethane or dichloromefhane/MeOH, followed by reduction with, for example, dimethyl sulfide to afford the carbonyl compound (LVI).
  • (LVI) may be reduced with sodium borohyd ⁇ de to afford the alcohol (LVIII) (where R 53 is O), which may be freated in with methanesulfonyl chloride in pyridine to afford the mesylate.
  • the methanesulfonate may be then freated with sodium azide in a solvent such as DMF at a temperature of from 25 °C to 120 °C to afford the azide, which may be reduced with, for example, palladium on barium sulfate and hydrogen gas to provide (LVIII) where R 53 is NH.
  • (LV) may be hydroborated with a borane reagent such as diborane followed by treatment with aqueous alkaline hydrogen peroxide to afford (LVII) where R 53 is O (LVII) may be manipulated at
  • R 53 m the manner described for (LVIII), and R 53 may be acylated alkylated, or sulfonylated as desired.
  • the R 52 group may be removed by treatment with sodium hydroxide m aqueous THF
  • Reaction scheme 12 depicts an alternate preparation of an intermediate of general formula (XXXV).
  • R 3 and R 4 are as defined for general formula (II).
  • RPG 2 is as defined for reaction scheme 6.
  • R 40 and R 41 are as defined for reaction scheme 7.
  • R 6 and R 4 7 are as defined for reaction scheme 9.
  • R ⁇ and R 4 7 may be taken together to constitute a three- to ten-membered ring.
  • R 54 and R 55 may be, independently, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocyclyl, aryl, heteroaryl, or hydrogen, where alkyl, alkenyl, alkynyl, cycloalkyl, and cycloalkenyl substituents may contain one or more O, S, SO, or S0 2 substituents.
  • the amine compound (LLX) is treated with (LX) in the presence of a tertiary base such as TEA or NMM to afford (LXI).
  • Treatment of (LXI) with silver nitrate and an amme HNR 6 R 4 7 provides (LXII). Removal of the alkyl group R 0 by sapomfication with aqueous base (or, if appropriate and where R 40 is tert-butyl, by freatment with frifluoroacetic acid) provides (XXXV).
  • Reaction scheme 13 describes an alternate preparation of an intermediate of formula
  • R 3 and R 4 are as defined for formula (II).
  • RPG 2 is as defined for reaction scheme 6.
  • R 40 is as defined for reaction scheme 7.
  • R 50 and R 5 ] are as defined for reaction scheme 12.
  • the intermediate of formula (LXIII) may be treated with osmium tetraoxide or similar derivative m the presence of an oxidizing agent such as an iron (III) salt in aqueous tert-butanol to afford (LXIV).
  • an oxidizing agent such as an iron (III) salt in aqueous tert-butanol to afford (LXIV).
  • Asymmetric induction may be achieved in the reaction via use of a chiral ligand, such as hydroqumine 1 ,4-phthalaz ⁇ ned ⁇ yl diether.
  • LXIV may be treated with thionyl chloride and a tertiary amme base and the resulting sulfite may be oxidized with Ru0 4 or RuCl 3 and NaI0 4 m aqueous acetonit ⁇ le/CCL, to afford the sulfate (LXV).
  • Reduction of the azide with palladium on carbon and hydrogen gives the amine (LXVII).
  • the amme may be alkylated with R 3 if desired, protected with RPG 2 (as described in previous schemes), and the R 40 ester group removed to provide (XXXV). Selection of R 40 as benzyl and RPG 2 as tert-butoxycarbonyl are desirable for the sequence of reactions m reaction scheme 13.
  • Reaction scheme 14 describes an alternate preparation of an intermediate of formula (XXXV).
  • R 3 and R are as defined for general formula (II).
  • RPG 2 is as defined for reaction scheme 6.
  • R40, - and R 2 are as defined for reaction scheme 7.
  • R54 is as defined for reaction scheme 12.
  • R 56 is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, heterocyclylene, arylene, O, NH, N-alkyl, or heteroarylene, where alkylene, alkenylene, alkynylene, cycloalkylene, and cycloalkenylene substituents may contain one or more O, S, SO, or S0 2 substituents.
  • R 57 is alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocyclyl, aryl, heteroaryl, or hydrogen, where alkyl, alkenyl, alkynyl, cycloalkyl, and cycloalkenyl substituents may contain one or more O, S, SO, or S0 2 substituents.
  • the amme compound (LIX) may be freated with the reagent R 57 -R 56 -R 2 -COCl m a solvent such as dichloromethane in the presence of tertiary base such as TEA to afford (LXVIII).
  • (LIX) may be freated with R 57 -R 56 - K -COOH (where R, 2 is not O, NH, or N-alkyl) and a dehydrating agent such as EDC in a solvent such as DMF or dichloromethane to afford (LXVIII).
  • the compound (LXVIII) where R 42 is NH may be prepared by treating (LIX) with R 57 -R 56 -NCO m a solvent such as dichloromethane. Removal of the alkyl group R 0 by sapomfication with aqueous base (or, if appropriate and where R 4 0 is tert-butyl, by treatment with frifluoroacetic acid) provides the acid (XXXV).
  • the compounds of the present invention can be administered m such oral (including buccal and sublmgual) dosage forms as tablets, capsules (each including timed release and sustained release formulations), pills, powders, granules, elixirs, tinctures, suspensions, syrups and emulsions. Likewise, they may also be administered m nasal, ophthalmic, otic, rectal, topical, intravenous (both bolus and infusion), mfrape ⁇ toneal, mtraarticular, subcutaneous or intramuscular inhalation or insufflation form, all using forms well known to those of ordinary skill m the pharmaceutical arts.
  • oral dosage forms as tablets, capsules (each including timed release and sustained release formulations), pills, powders, granules, elixirs, tinctures, suspensions, syrups and emulsions.
  • they may also be administered m nasal, ophthalmic, otic, rectal, topical, intravenous (both bolus and
  • the dosage regimen utilizing the compounds of the present invention is selected accordance with a variety of factors including type, species, age, weight, sex and medical condition of the patient; the severity of the condition to be treated; the route of administration: the renal and hepatic function of the patient; and the particular compound or salt thereof employed.
  • An ordinarily skilled physician or veterinarian can readily determine and prescribe the effective amount of the drug required to prevent, counter or arrest the progress of the condition.
  • Oral dosages of the present invention when used for the indicated effects, will range between about 0.1 to 2000 mg kg of body weight per day, and particularly 1 to 100 mg/kg of body weight per day. Oral dosage units will generally be administered in the range of from 1 to about 250 mg and more preferably from about 25 to 250 mg The daily dosage for a 70 kg mammal will generally be in the range of about 10 mg to 5 grams of a compound of formula (I) or (II).
  • While the dosage to be administered is based on the usual conditions such as the physical condition of the patient, age, body weight, past medical history, route of administrations, severity of the conditions and the like, it is generally preferred for oral administration to administer to a human. In some cases, a lower dose is sufficient and, in some cases, a higher dose or more doses may be necessary. Topical application similarly may be once or more than once per day depending upon the usual medical considerations.
  • compounds of the present invention may be administered in a single daily dose, or the total daily dosage may be administered in divided doses of two, three or four times daily.
  • preferred compounds for the present invention can be administered in infranasal form via topical use of suitable mtranasal vehicles, or via transdermal routes, using those forms of transdermal skin patches well known to those of ordinary skill in that art.
  • the dosage administration will, of course, be continuous rather than intermittent throughout the dosage regimen.
  • the compounds herein described m detail can form the active ingredient, and are typically administered in admixture with suitable pharmaceutical diluents, excipients or carriers (collectively referred to herein as "earner" materials) suitably selected with respect to the intended form of administration, that is, oral tablets, capsules, elixirs, syrups and the like, and consistent with conventional pharmaceutical practices.
  • the active drug component can be combined with an oral, non-toxic pharmaceutically acceptable inert earner such as ethanol, glycerol, water and the like Powders are prepared by comminuting the compound to a suitable fine size and mixing with a similarly comminuted pharmaceutical carrier such as an edible carbohydrate, as, for example, starch or manmtol. Flavoring, preservative, dispersing and coloring agent can also be present.
  • an oral, non-toxic pharmaceutically acceptable inert earner such as ethanol, glycerol, water and the like
  • Powders are prepared by comminuting the compound to a suitable fine size and mixing with a similarly comminuted pharmaceutical carrier such as an edible carbohydrate, as, for example, starch or manmtol. Flavoring, preservative, dispersing and coloring agent can also be present.
  • Capsules are made by preparing a powder mixture as described above, and filling formed gelatin sheaths.
  • Ghdants and lubricants such as colloidal silica, talc, magnesium stearate, calcium stearate or solid polyethylene glycol can be added to the powder mixture before the filling operation.
  • a disintegrating or solubihzing agent such as agar-agar, calcium carbonate or sodium carbonate can also be added to improve the availability of the medicament when the capsule is ingested.
  • suitable binders include starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, fragacanth or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes and the like.
  • Lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like.
  • Disintegrators include, without limitation, starch, methyl cellulose, agar, bentonite, xanthan gum and the like.
  • Tablets are formulated, for example, by preparing a powder mixture, granulating or slugging, adding a lubricant and dis tegrant and pressing into tablets.
  • a powder mixture is prepared by mixing the compound, suitably comminuted, with a diluent or base as described above, and optionally, with a binder such as carboxymethylcellulose, an ahginate, gelatin, or polyvmyl pyrrolidone, a solution retardant such as paraffin, a resorption accelerator such as a quaternary salt and/or an absorption agent such as bentonite, kaolm or dicalcium phosphate
  • the powder mixture can be granulated by wetting with a binder such as syrup, starch paste, acadia mucilage or solutions of cellulosic or polymeric materials and forcing through a screen.
  • the powder mixture can be run through the tablet machine and the result is imperfectly formed slugs broken into granules.
  • the granules can be lubricated to prevent sticking to the tablet forming dies by means of the addition of stearic acid, a stearate salt, talc or mineral oil.
  • the lubricated mixture is then compressed into tablets.
  • the compounds of the present invention can also be combined with free flowing inert carrier and compressed into tablets directly without going through the granulating or slugging steps
  • a clear or opaque protective coating consisting of a sealing coat of shellac, a coating of sugar or polymeric material and a polish coating of wax can be provided. Dyestuffs can be added to these coatings to distinguish different unit dosages.
  • Oral fluids such as solution, syrups and elixirs can be prepared in dosage unit form so that a given quantity contains a predetermined amount of the compound Syrups can be prepared by dissolving the compound in a suitably flavored aqueous solution, while elixirs are prepared through the use of a non-toxic alcoholic vehicle.
  • Suspensions can be formulated by dispersing the compound in a non-toxic vehicle.
  • Solubihzers and emulsifiers such as ethoxylated isostearyl alcohols and polyoxy ethylene sorbitol ethers, preservatives, flavor additive such as peppermint oil or saccharin, and the like can also be added.
  • dosage unit formulations for oral administration can be microencapsulated.
  • the formulation can also be prepared to prolong or sustain the release as for example by coating or embedding particulate material polymers, wax or the like.
  • the compounds of the present invention can also be administered in the form of l posome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles.
  • Liposomes can be formed from a variety of phosphohpids, such as cholesterol, stearylamine or phosphatidylcholmes.
  • Compounds of the present invention may also be delivered by the use of monoclonal antibodies as individual carriers to which the compound molecules are coupled.
  • the compounds of the present invention may also be coupled with soluble polymers as targetable drug carriers.
  • Such polymers can include polyvinylpyrrohdone, pyran copolymer, polyhydroxypropylmethacrylamide-phenol, polyhydroxyethylaspartamidephenol, or polyethyleneoxidepolylysme substituted with palmitoyl residues.
  • the compounds of the present invention may be coupled to a class of biodegradable polymers useful m achieving controlled release of a drug, for example, polylactic acid, polepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates and cross-linked or amphipafhic block copolymers of hydrogels.
  • a class of biodegradable polymers useful m achieving controlled release of a drug for example, polylactic acid, polepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates and cross-linked or amphipafhic block copolymers of hydrogels.
  • the present invention includes pharmaceutical compositions containing 0.1 to 99.5%, more particularly, 0.5 to 90% of a compound of the formula (II) in combination with a pharmaceutically acceptable carrier.
  • Parenteral administration can be effected by utilizing liquid dosage unit forms such as sterile solutions and suspensions intended for subcutaneous, intramuscular, lntrathecal, mfraarte ⁇ al or intravenous injection. These are prepared by suspending or dissolving a measured amount of the compound in a non-toxic liquid vehicle suitable for injection such as aqueous oleaginous medium and sterilizing the suspension or solution. Alternatively, a measured amount of the compound is placed m a vial and the vial and its contents are sterilized and sealed. An accompanying vial or vehicle can be provided for mixing prior to administration. Non-toxic salts and salt solutions can be added to render the injection lsotonic. Stabilizers, preservations and emulsifiers can also be added.
  • sterile solutions and suspensions intended for subcutaneous, intramuscular, lntrathecal, mfraarte ⁇ al or intravenous injection. These are prepared by suspending or dissolving a measured amount of the compound in a non
  • Rectal administration can be effected utilizing suppositories which the compound is admixed with low-meltmg water-soluble or insoluble solids such as polyethylene glycol, cocoa butter, higher ester as for example flavored aqueous solution, while elixirs are prepared through my ⁇ styl palmitate or mixtures thereof.
  • low-meltmg water-soluble or insoluble solids such as polyethylene glycol, cocoa butter, higher ester as for example flavored aqueous solution, while elixirs are prepared through my ⁇ styl palmitate or mixtures thereof.
  • Topical formulations of the present invention may be presented as, for instance, ointments, creams or lotions, eye ointments and eye or ear drops, impregnated dressings and aerosols, and may contain appropriate conventional additives such as preservatives, solvents to assist drug penetration and emollients in ointments and creams.
  • the formulations may also contain compatible conventional carriers, such as cream or ointment bases and ethanol or oleyl alcohol for lotions. Such carriers may be present as from about 1% up to about 98% of the formulation More usually they will form up to about 80% of the formulation.
  • the compounds according to the invention are conveniently delivered m the form of an aerosol spray presentation from pressurized packs or a nebulizer, with the use of a suitable propellant, e.g. dichlorodifluoromethane, t ⁇ chlorofluoromefhane, dichlorotetrafluoroethane, tetrafluoroethane, heptafluoropropane, carbon dioxide or other suitable gas.
  • a suitable propellant e.g. dichlorodifluoromethane, t ⁇ chlorofluoromefhane, dichlorotetrafluoroethane, tetrafluoroethane, heptafluoropropane, carbon dioxide or other suitable gas.
  • a suitable propellant e.g. dichlorodifluoromethane, t ⁇ chlorofluoromefhane, dichlorotetrafluoroethane, tetrafluoroethane, h
  • gelatin for use in an inhaler or insufflator may be formulated containing a powder mix of a compound of the invention and a suitable powder base such as lactose or starch.
  • suitable powder base such as lactose or starch.
  • the prefe ⁇ ed pharmaceutical compositions are those in a form suitable for oral administration, such as tablets and liquids and the like and topical formulations.
  • the resulting solution is allowed to warm to 23 °C and is stirred at 23 °C for 14 h, concentrated in vacuo. and diluted with dichloromethane (500 mL). The organic layer is washed with 1 N hydrochloric acid, saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, and concenfrated to provide the methanesulfonate intermediate.
  • reaction mixture is concentrated, diluted with EtOAc (20 mL) and washed sequentially with 1 M aqueous sodium carbonate and saturated aqueous sodium chloride.
  • the organic layer is dried over magnesium sulfate, concentrated in vacuo, and purified by silica gel chromatography (elution with 2.1 EtOAc - hexane) to provide (2R,3S)-3-(formyl-2- tetrahydropyranyloxyammo)-2-(3-phenyl-l-propyl)butano ⁇ c acid [(lS)-2,2-d ⁇ methyl-l-(3- py ⁇ dylcarbamoyl)-l-propyl]am ⁇ de as a solid (115 mg, 62% yield) ESI-MS m/z 539 (M+H) + .
  • Example 2b 4-Phenylcyclohexylmethanol.
  • a solution of 4-phenyl-l -methylenecyclohexane (8.9 g, 0.051 mol) in THF (10 mL) is added dropwise to a solution of borane (15 mL, 0.015 mol, 1 M m THF) at 0 °C under argon. The mixture is stirred at 25 °C for 2.5 h.
  • a solution of aqueous sodium hydroxide (5 mL, 0.015 mol, 3 M) is added dropwise, the mixture is chilled to 0 °C and aqueous hydrogen peroxide (6 mL, 0.06 mol, 30%) is added dropwise. After stirring at 25 °C for 1 h the mixture is poured into water and extracted with ether.
  • Imidazole (4.4 g, 0.065 mol) is added in one portion to a solution of t ⁇ phenylphosphme (17.0 g, 0.065 mol) m dichloromethane (300 mL) at 25 °C . After all of the solids dissolved, iodine (16.5 g, 0.065 mol) is added in four portions. The mixture is stirred for 10 min, then a solution of 4-phenylcyclohexylmethanol (10.5 g, 0.055 mol) is added dropwise and the mixture is stirred at 25 °C for 18 h. The reaction mixture is poured into pentane and the resulting solids filtered.
  • Example 2d Methyl (2R,3R)-3-Hydroxy-2-(4-phenylcyclohexylmethyl)butanoate
  • a solution of methyl (3R)-3-hydroxybutyrate (5.6 g, 0.047 mol) in THF (20 mL) is added dropwise to a solution of LDA prepared by dropwise addition of n-butylhthium (49.5 mL, 0.099 mol, 2 M in cyclohexane) to dnsopropylamme (10 0 g, 0.099 mol) in THF (20 mL) at -78 °C.
  • lithium hydroxide monohydrate (0.59 g, 0.014 mol) and lithium hydroxide monohydrate (0.59 g, 0.014 mol) is stirred at 25 °C m a mixture of THF (30 mL), methanol (15 mL) and water (15 mL) for 72 h. The reaction is not complete and an additional 0.88 g (0.021 mol) of lithium hydroxide monohydrate m water (10 mL) is added. After 6 h an additional 0.59 g (0.014 mol) of solid lithium hydroxide monohydrate and the mixture is stirred at 25 °C for 18 h. The mixture is concentrated in vacuo and the residue is taken up in water, chilled in an ice-bath and the pH is adjusted to 2 with sodium bisulfate.
  • Methanesulfonyl chloride (1.4 g, 0.012 mol) is added dropwise to a solution of
  • a solution of 3 N sodium hydroxide (18.7 mL) is added dropwise to a solution of (3R,4S)-1- (2-tefrahydropyranyloxy)-3-(4-phenylcyclohexylmethyl)-4-methylazet ⁇ dm-2-one (3.6 g, 0.010 mol) m dioxane (50 mL) at 25 °C and the mixture is stirred for 18 h.
  • Water (10 mL) is added to the reaction mixture and an additional 5 mL of 3 N sodium hydroxide is added dropwise and the mixture is stirred an additional 8 h.
  • Formic acetic anhydride (2.03 g, 0.023 mol) is added via syringe to a mixture of (2R,3S)-2-(4-phenylcyclohexylmethyl)-3-(2-tefrahydropyranyloxyammo)butano ⁇ c acid (3.44 g, 0.009 mol) in pyridme (35 mL) at 0 °C. The mixture is allowed to come to 25 °C. Additional formic acetic anhydride (0.97g, 0.011 mol) is added. The mixture is stirred at 25 °C for 8 h and concentrated in vacuo .
  • Methyl (3R)-3-hydroxyhexanoate is prepared m the same manner described above in methanol employing methyl butyrylacetate as the starting ketoester.
  • the enantiomenc excess is 99% as determined by chiral analytical HPLC methods.
  • Example 3c Ethyl (2R,3R)-2-(3-Phenyl-l-propyl)-3-hydroxyhexanoate
  • a solution of ethyl (2R,3R)-2-[(2£)-3-phenyl-2-propen-l-yl]-3- hydroxyhexanoate (1.76 g, 6.38 mmol) in 30 mL of EtOAc is freated with 200 mg of 10% palladium on carbon.
  • the resulting solution is heated to 50 °C and is stirred for 18 h.
  • the reaction mixture is cooled to 25 °C, concentrated in vacuo, diluted with 20 mL of EtOAc, and washed with 1 M aqueous sodium carbonate solution.
  • reaction mixture is quenched by addition 10 mL of saturated aqueous ammonium chloride solution, is poured into 100 mL of 1 M hydrochloric acid, and is extracted with two 100 mL portions of EtOAc. The combined organic layers are dried over magnesium sulfate, concentrated in vacuo, and purified by silica gel chromatography (elution with 20% EtOAc - hexanes) to afford methyl
  • reaction mixture is purified by silica gel chromatography (elution with 8: 1 hexanes - EtOAc) to provide (3R,4S)-l-(2-tefrahydropyranyloxy)-3-((2E)-3- t ⁇ butylstannyl-2-propene-l-yl)-4-methylazet ⁇ d ⁇ n-2-one as an oil (2.6 g, 87% yield) ⁇
  • NMR 400 MHz, CDCI3 ⁇ 5.94 (m, 2H), 5.16 (m, 0.5H), 4.99 (m, 0.5H), 4.20-4.01 (m,
  • the organic layer is dried over anhydrous magnesium sulfate and concenfrated under reduced pressure to provide (2R,3S)-2-(3-(4- chlorophenyl)-l-propyl)-3-(formyl-2-tetrahydropyranyloxyam ⁇ no)butano ⁇ c acid as an oil (385 mg, 94% yield).
  • Assays are conducted m a total volume of 0.180 mL assay buffer (200 mM NaCl, 50 mM T ⁇ s, 5 mM CaCl 2 , 10 ⁇ M ZnS0 , 0.005% B ⁇ j 35, pH 7.6) in each
  • the potency of compounds of the invention as inhibitors of cell - free tumor necrosis factor ⁇ converting enzyme is determined as follows; Membrane preparation from MonoMac 6 cells (subfractionated extract from equivalent of 6x10 6 cells per 60 ⁇ l assay) is incubated for 1 h with 200 nM radiolabeled substrate (B ⁇ otm-SPLAQAVRSSSRT-( 3 H)P-S-NH 2 , 4.1 Ci/mmol, ref # 0935 from Zeneca) in 10 mM hepes buffer, 250 mM sucrose, pH 7.5. The reaction is quenched by addition of streptavidin SPA beads (Amersham RPNQ0006), with excess binding capacity relative to substrate, suspended in 250 mM EDTA, pH 8.0.
  • Binding is complete within 15 minutes and plates are counted m a Wallac 1450 Microbeta liquid scintillation counter. Percent inhibition is calculated at each inhibitor concentration and the data were plotted using standard curve fitting programs. IC 50 values were determined from these curves. Assays were run at low substrate concentration ([S] «K m ) such that the calculated IC 50 values are equivalent to K, within experimental error.
  • the potency of compounds of the invention as inhibitors of release of soluble tumor necrosis factor ⁇ from stimulated monocytes in vitro is determined as follows; LPS/PMA solution for assay consisting of a) 4 ⁇ L of 5 mg/mL LPS stock and b) 6 ⁇ L of 10 mg/mL
  • PMA stock are added to 500 ⁇ L of medium (RPMI 1640 (Gibco) + 10% FBS + penicillin/streptomycin + 1-glutam ⁇ ne). This solution is then diluted 1.1000 (40 ng/mL and 120 ng/mL) for use later in the assay.
  • Compounds (10 mM) are serially diluted 1 :3 in DMSO.
  • MonoMac 6 cell suspension (130 ⁇ L, 1.5 xlO 6 cells/mL) is then added to each well resulting m 2 x 10 5 cells/well.
  • LPS/PMA (50 ⁇ L) solution is then added to each well to begin stimulation (final concentrations of 10 ng/mL and 30 ng/mL respectively)
  • the plate is incubated at 37 °C for 2 hours then spun at 1,500 rpm for 3 minutes to pellet cells.
  • the supernatant 120 ⁇ L/well
  • 20 ⁇ L of the supernatant is transferred to a Cistron TNF ⁇ ELISA plate and processed according to the manufacturer's instructions to quantitate levels of TNF ⁇ . Percent inhibition of TNF ⁇ release is calculated at each inhibitor concentration and the data were plotted using standard curve fitting programs. IC 50 values were determined from these curves.
  • the potency of compounds of the invention as inhibitors of serum TNF ⁇ elevation in mice treated with lipopolysaccharide (LPS) is determined as follows; a) for subcutaneous (s.c.) administration, test compound is dissolved in DMSO and added to a mixture of 0.9% sodium chloride solution and 30% Trappsol HPB-20 (Cyclodextrin Technology Development Inc., Gainesville, Florida USA) for a final DMSO concentration of 1%.
  • test compounds are formulated in 0.2 mL of PBS and 0.1% Tween 80 and given orally via gavage 10 mm prior to LPS administration.
  • C3/hen female mice are injected mfrape ⁇ toneally with 200 ⁇ g/kg LPS (Escherichia coli, Serotype 0111 :B4, Sigma Chemical Co, St. Louis, MO) in PBS and sacrificed 90 mm later by C0 2 asphyxiation. Blood is immediately taken from the caudal vena cava and plasma prepared and frozen at -80 °C. Plasma concenfrations of TNF are measured by ELISA (Genzyme Co., Cambridge MA). Results are listed in Table 4.

Abstract

A family of compounds having general structural formula (I) where W is a reverse hydroxamic acid group, and R1, R2, R3, R4, R5 and R6 are as described in the specification, or a pharmaceutically acceptable salt, solvate, biohydrolyzable ester, biohydrolyzable amide, affinity reagent, or prodrug thereof. Also described are methods for their preparation, pharmaceutical compositions including such compounds and their use in medicine.

Description

FORMAMIDE COMPOUNDS AS THERAPEUTIC AGENTS
FIELD OF THE INVENTION
The present invention provides novel compounds, novel compositions, methods of their use and methods of their manufacture, such compounds generally pharmacologically useful as agents in those disease states alleviated by the inhibition or antagonism of matrix metallopro teases, metalloproteases, and/or tumor necrosis factor-alpha (TNF), which pathologically involve aberrant extracellular matrix degradation, shedding of cell surface protein ectodomains, and/or TNF synthesis, such disease states including arthritis, tumor metastasis and diabetes. The aforementioned pharmacologic activities are useful in the treatment of mammals.
More specifically, the compounds of the present invention can be used in the treatment of rheumatoid arthritis, osteoarthritis, inflammatory bowel syndromes, periodontal disease, aberrant angiogenesis, tumor invasion and metastasis, comeal ulceration and the complications of diabetes. At the present time, there is a need in the areas of rheumatology, oncology, dentistry, opththalmology, gastroenterology, cardiology, neurology, nephrology, infectious disease and endocrinology therapy for such agents.
BACKGROUND OF THE INVENTION
The matrix metalloprotease (MMP) family of zinc endoproteases includes fibroblast collagenase (MMP-1, collagenase-1), neutrophil collagenase (MMP-8, collagenase-2), chondrocyte collagenase (MMP-13, collagenase-3), gelatinases A and B (MMP's 2 and 9), and members of the stromelysin family such as stromelysin-1 (MMP-3), stromelysin-3 (MMP-11), and matrilysm (MMP-7). These enzymes accelerate breakdown of connective tissue by catalyzed resorption of the extracellular matrix. This is a feature of diverse pathologies; therefore, inhibitors of one or more of the matrix metalloproteases would have utility m a wide range of disease states such as in abrogating the initiation of tumor metastasis and angiogenesis and in halting the pathogenesis of demyelmating diseases of the nervous system, multiple sclerosis being one example MMP inhibitors would also find utility in diseases involving connective tissue degradation in the joint, as occurs in osteoarthritis and rheumatoid arthritis. MMP's-1 and -3 have been found in elevated levels in the synovial fluid of patients with rheumatoid arthritis and osteoarthritis.
Collagenase-3 (MMP-13) is a member of the family of MMP's which preferentially digest collagen. Collagenase-3 is one of the more newly characterized MMP's; biochemical studies on the recombinant protein have demonstrated that it cleaves type II collagen, the predominant matrix component of articular cartilage, more efficiently than either MMP- 1 or MMP -2 and that it is expressed by chondrocytes in osteoarthritic cartilage These data would implicate collagenase-3 as a significant target in rheumatoid arthritis and osteoarthritis for inhibition by MMP inhibitors.
Compounds which inhibit the activities of one or more of the matrix metalloproteases are recognized as having therapeutic benefit m one or more pathologies where MMP activity is upregulated, such as;
I) inflammatory/autoimmune diseases, including but not limited to rheumatoid arthritis, osteoarthritis, Crohn's disease and other inflammatory bowel diseases, periodontal disease, gingivitis, and comeal ulceration; n) cardiovascular diseases, including but not limited to atherosclerosis, and restenosis; in) metabolic diseases, including but not limited to complications of diabetes, osteoporosis, and other disorders involving resorption of bone; IV) neurologic diseases, including but not limited to multiple sclerosis and other demyehnation ailments; v) diseases of cancer and malignancy, including but not limited to cancers of the oral cavity and pharynx (lip, tongue, mouth, pharynx), esophagus, stomach, small intestine, large intestine, rectum, liver and biliary passages, pancreas, larynx, lung, bone, connective tissue, skin, colon, breast, cervix uteri, corpus endometπum, ovary, prostate, testis, bladder, kidney and other urinary tissues, eye, brain and central nervous system, thyroid and other endocrine gland, leukemias (lymphocytic, granulocytic, monocytic), Hodgkm's disease, non-Hodgkin's lymphomas, multiple myeloma, tumor invasion, and metastatic and angiogemc events thereof; vi) renal diseases, including but not limited to nephrotic syndromes and glomerulonephritis; vn) infectious diseases, including but not limited to those mediated by viruses, bacteria, and fungi; and vm) respiratory diseases, including but not limited to emphysema and COPD.
Many inhibitors of matrix metalloproteases have been disclosed, including some structure activity relationships for a series of carboxylalkylamrne inhibitors. These molecules are exemplary for MMP inhibitors in general. They generally embody a functional group capable of tightly binding the zinc cofactor at the enzyme active site, which is contained within a peptidic or pseudopeptide structure. Zinc binding groups among the MMP inhibitor art have included hydroxamic acid, reverse hydroxamic acid, thiol, carboxylate, and phosphmate.
Hydroxamate metalloprotease inhibitors disclosed in the art usually have the following general structure (I):
Figure imgf000005_0001
(I) where W is a zmc-chelatmg acyl derivative group of the formula -C(0)NHOH (which by convention and in this application are referred to as "forward hydroxamates") or a zmc- chelatmg substituted amine group of the formula -NH(OH)C(0)R (which by convention and in this application are referred to as "reverse hydroxamates"), where R is usually hydrogen or alkyl The other substituents vary according to specifications expressed by the art disclosure. It is understood and demonstrated that variations in these substituents can have dramatic effects on potency and selectivities between the matrix metalloproteases.
Suppression of MMP activity in conditions characterized by its overproduction would be of benefit, and compounds which inhibit MMP's would act in this manner at a specific target and be useful and of benefit. The present invention fills this need by providing compounds that are potetn, specific, orally active inhibitors of matrix metalloproteases.
Tumor necrosis factor-α (TNFα), hereinafter called "TNF", is a mammalian protein capable of inducing cellular effects by virtue of its interaction with specific cellular receptors.
It was initially characterized and so named due to its ability to cause death of cancerous cells
It is produced primarily by activated monocytes and macrophages. Human TNF is produced as a larger pro- form of 26 kD which is processed to a secreted 17 kD mature form by proteolytic processing of the alamne-76 - valme-77 peptide bond.
Recently, certain compounds having matrix metalloprotease - inhibiting activity have been found to inhibit the release of mature 17 kD TNF from cells. Further, these inhibitors also protect mice from a lethal dose of endotoxin indicating that the compounds can inhibit
TNF secretion in vivo. These compounds inhibit the cell-associated proteolytic processing of the 26 kD pro-TNF to the mature 17 kD form. The proteolytic activity is thought to reside in an mtracellular or cell-associated specific enzyme or family of enzymes, which by convention is called a "TNF convertase", distinct from the matrix metalloproteases but related in that both contain a zmc cation at the active site TNF convertase enzymatic activity can be detected m monocyte membrane fractions, and the enzyme activity can be inhibited by certain matrix metalloprotease - inhibiting compounds.
A metalloprotease is thought to mediate the proteolysis of the cell - surface IgE receptor CD23. Certain of the CD23 - derived peptides possess biological activitites mimicking those of cytokmes, including TNFα.
Metalloprotease - like activity is also thought to contribute to the shedding of certain cell surface protein ectodomains such as L-selectm, fibronectin, thyrotropin stimulating hormone receptor, transforming growth factor alpha precursor, low density hpoprotein receptor, beta amyloid precursor protein, mterleukm-6 receptor alpha subunit, Fas ligand, CD40 ligand, epidermal growth factor receptor, macrophage colony stimulating factor, mterleukιn-1 receptor type II, CD30, and tumor necrosis factor receptors type I and II.
TNF is known to mediate many biological responses in vivo. Preclimcal and clinical studies in animals and humans with specific TNF neutralizing antibodies, soluble TNF receptor constructs, and TNF detection techniques have implicated TNF as a mediator m numerous pathologies. The compounds of the present invention by virtue of their activity in inhibiting TNF production and/or their activity in preventing cell surface protein ectodomain shedding should show utility in the treatment of diverse pathologies such as; I) inflammatory/autoimmune diseases, including but not limited to rheumatoid arthritis, osteoarthritis, Crohn's disease and other inflammatory bowel diseases and inflammatory gastrointestinal diseases, and systemic lupus erythematosis, n) reperfusion injuries, such as those caused by an initial ischemic event; m) systemic inflammatory response syndromes, including but not limited to sepsis, burn injury, pancreatitis, and adult respiratory distress syndrome; IV) allergic and dermatologic diseases, including but not limited to delayed type hypersensitivity, psoriasis, asthma, eczema, allergic rhinitis, and allergic conjunctivitis; v) cardiovascular diseases, including but not limited to hyperhpidemia, myocardial infarction. atherosclerosis, chronic obstructive pulmonary disease, and restenosis; vi) metabolic diseases, including but not limited to osteoporosis, obesity, and diabetes; vn) neurologic diseases, including but not limited to Alzheimer's disease, Parkinson's disease, multiple sclerosis, aneurism, and stroke; vm) transplant rejection, including but not limited to organ transplant rejection and graft versus host disease; ix) diseases of cancer and malignancy, including but not limited to cancers of the oral cavity and pharynx (lip, tongue, mouth, pharynx), esophagus, stomach, small intestine, large intestine, rectum, liver and biliary passages, pancreas, larynx, lung, bone, connective tissue, skm, colon, breast, cervix uteri, corpus endometπum, ovary, prostate, testis, bladder, kidney and other urinary tissues, eye, brain and central nervous system, thyroid and other endocπne gland, leukemias (lymphocytic, granulocytic, monocytic), Hodgkm's disease, non-Hodgkm's lymphomas, multiple myeloma, tumor invasion, and metastatic and angiogemc events thereof; x) renal diseases, including but not limited to nephrotic syndromes and glomerulonephπtis; xi) cachexia and related wasting syndromes; xn) infectious diseases, including but not limited to HIV infection and neuropathy, Epstem- Barr viral infection, herpes viral infection, malaria, meningitis, schistosomiasis, leprosy, hepatitis (which includes hepatitis A, hepatitis B, and hepatitis C), infectious arthritis, leishmaniasis, tuberculosis, Lyme disease, and viral encephalitis; xin) effects of disease therapy, including but not limited to cytokine therapy, chemotherapy, radiation therapy and therapies using anti-T-cell antibodies or cytotoxin-antibody conjugates, and xiv) ocular diseases, including but not limited to diabetic retinopathy and macular degeneration. Suppression of TNF activity in conditions characterized by its overproduction would be of benefit, and compounds which inhibit TNF convertase would act m this manner at a specific target and be useful and of benefit. The present invention fulfills this need by providing potent, specific, orally active inhibitors of TNF-alpha release from monocyte cells acting via inhibition of TNF-alpha converting enzyme (TNFc).
Suppression of shedding of cell surface protein ectodomains m conditions characterized by an overactivity of such a shedding enzyme or enzymes would be of benefit, and compounds which inhibit this cell surface protein ectodomam shedding would be useful and of benefit. The present invention fulfills this need by providing potent, orally active inhibitors of shedding of cell surface protein ectodomains acting via inhibition of one or more specific enzymes which mediate this proteolytic event.
Suppression of CD23 proteolysis in conditions characterized by an overabundance of CD23 proteolytic fragments would be of benefit, and compounds which inhibit CD23 proteolysis would be useful and of benefit. The present invention fills this need by providing potent inhibitors of CD23 proteolysis acting via inhibition of one or more specific enzymes which mediate this proteolytic event.
SUMMARY OF THE INVENTION
Accordingly it is an object of the present invention to provide a potent, specific, orally active inhibitor of matrix metalloproteases It is another object of the present invention to provide a potent, specific, orally active inhibitor of TNF-alpha release from monocyte cells acting via inhibition of TNF-alpha converting enzyme (TNFc).
Accordingly it is another object of the present invention to provide a potent, orally active inhibitor of shedding of cell surface protein ectodomains acting via inhibition of one or more specific enzymes which mediate this proteolytic event.
Accordingly it is another object of the present invention to provide a potent inhibitor of CD23 proteolysis acting via inhibition of one or more specific enzymes which mediate this proteolytic event.
It is an object, therefore, of the present invention to provide a compound of the formula
Figure imgf000010_0001
(li) where Ri is
.A
,A,
whereA) is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, or a direct bond;
A2 is C(0)NR7, NR7C(0), S02NR7, NR7S02,NR7, S, SO, S02, O, or a direct bond, where R7 is as defined below; A3 is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, or a direct bond;
A4 is alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocyclyl, heteroaryl, aryl, NR8R9,
OR8, or hydrogen, where R8 and R9 are as defined below;
R2 ιs
Figure imgf000011_0001
whereD, is alkylene, alkenylene, alkynylene, NR10(O)C, NRι0, S, SO, S02, O, or a direct bond, where R]0 is as defined below;
D2 is S, SO, S02, O, C(0)NRu, NR„C(0), NR,,, or a direct bond, where Rn is as defined below;
D3 isalkylene, alkenylene, alkynylene, arylene, heteroarylene, S, SO, S02, O, C(0)NRι2,
NR12C(0), S02NR12, NR12S02, NR12, or a direct bond, where R12 is as defined below;
D4 is aryl, aryloxy, heteroaryl, or heteroaryloxy;
D5 and D6 are, independently, lower alkylene, O, S, SO, or S02; R3 is hydrogen or lower alkyl;
R4 is
ΛΛΛΛΛΛ
E' E1
-E,
E-E5
I 6
where E] is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, C(0)NRπ, or a direct bond, where R]3 is as defined below; E is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, NRU, S, SO, S02, O, C(O), or a direct bond, where R14 is as defined below;
E3 is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, NR15, S, SO, S02, O, C(O),
Figure imgf000012_0001
or a direct bond, where R)5 and R16 are as defined below;
E4 is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, NR18, S, SO, SO,, O, N(R18)C(0), C(0)N(R18), C(O),
Figure imgf000012_0002
or a direct bond, where R]7 and R!8 are as defined below;
E5 is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, NR,9, S, SO, S02, O, C(O),
Figure imgf000012_0003
or a direct bond, where R]9 and R20 are as defined below;
E6 is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, NR2ι, S, SO, S02, O, C(O), or a direct bond, where R2! is as defined below; E7 is hydrogen, NR22R23, OR22, SR22, SOR22, S02R22, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocyclyl, aryl, or heteroaryl, where R22 and R23 are as defined below; R5 is hydrogen or lower alkyl; Re is where Zi is heteroarylene;
Z2 is lower alkylene, lower alkenylene, lower alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, C(0)NR24, NR24C(0), S02NR24, NR24S02, NR24, S, SO, S02, O, C(O), C(0)0, OC(O), or a direct bond, where R24 is as defined below;
Z3 is alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocyclyl, heteroaryl, aryl, NR25R26, OR25, or hydrogen, where R25 and R26 are as defined below; and
R7, Rg, R9, Rio, Ri b Rl2; Rl3> Rl4, Rl5> Rl6) Rl7> Rl8, l9, R20, R2I ) ∑2> R23> R24, R25) and R26 are, independently, hydrogen, alkyl, alkynyl, alkenyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, or heteroaryl; or a pharmaceutically acceptable salt, solvate, biohydrolyzable ester, biohydrolyzable amide, affinity reagent, or prodrug thereof.
DETAILED DESCRIPTION OF THE INVENTION
The present invention provides a family of compounds having the general structural formula:
Figure imgf000014_0001
(I)
or a pharmaceutically acceptable salt, solvate, biohydrolyzable esters, biohydrolyzable amides, affinity reagents, or prodrugs thereof, wherein
W is a reverse hydroxamic acid group;
Ri is a substituent other than hydrogen; i is a hpophilic substituent preferably with steπc bulk proximal to the peptide backbone, and; R6 is a heteroaryl substituent.
Such compounds are novel and are unknown in the art and, given the appropriate choice of Ri, R2, R3, t, R5, and R<;, as described herein, show potent inhibition of MMP's, cell-free TNF convertase enzyme and TNF release from cells, and m some cases inhibit TNF convertase and TNF release from cells in preference to matrix metalloproteases. The heteroaryl nature of Pv, in combination with an appropriate choice of R1 ; R2, R3, R4, and R5 as described herein is beneficial in achieving increased potency against TNF release from cells relative to inhibition of MMP's Such molecules can be selective for TNF inhibition over MMP's and can possess an improved therapeutic profile where inhibition of one or more of the matrix metalloproteases is associated with an adverse biological response or abnormal pathology. The heteroaryl nature of R<; in combination with an appropriate choice of Ri, R ,
R3, R_t, and R5 as described herein is also beneficial in achieving selective inhibition of one or more of the matrix metalloproteases (for example, collagenase-3) in preference to TNF convertase inhibition and inhibition of TNF release from whole cells.
In particular, a preferred group of compounds of the present invention include those of the formula (II)'
Figure imgf000015_0001
(II) where R, is
Figure imgf000015_0002
where A) is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, or a direct bond;
A2 is
C(0)NR7, NR7C(0), SO2NR7, NR7SO2 NR7, S, SO, S02, O, or a direct bond, where R7 is as defined below;
A3 is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, or a direct bond;
At is alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocyclyl, heteroaryl, aryl, NR8R9, OR8 or hydrogen, where R8 and R9 are as defined below;
R, ιs
Figure imgf000016_0001
where
alkylene, alkenylene, alkynylene, NR10(O)C, NR10, S, SO, S02, O, or a direct bond, where Rio is as defined below;
D2 is
S, SO, S02, O, C(0)NR, ι, NR, ,C(0), NR„, or a direct bond, where R, , is as defined below;
D3 ιs alkylene, alkenylene, alkynylene, cycloalkylene, arylene, heteroarylene, S, SO, S02, O, C(0)NR,2, NR,2C(0), S02NR12, NR,2S02, NR,2, or a direct bond, where R,2 is as defined below;
D4 is aryl, aryloxy, heteroaryl, or heteroaryloxy,
D5 and D6 are, independently, lower alkylene, O, S, SO, or S02;
R3 is hydrogen or lower alkyl;
E , 2' E1
^ ι β
E,
where Ei is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, C(0)NRι3, or a direct bond, where Rπ is as defined below;
E2 is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, NR,4, S, SO, S02, O, C(O), or a direct bond, where R is as defined below;
E3 is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, NR15, S, SO, S02, O, C(O),
Figure imgf000017_0001
or a direct bond, where R^ and Rι6 are as defined below; E4 is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, NR,8, S, SO, S02, O, N(R,8)C(0), C(0)N(R18), C(O),
Figure imgf000018_0001
or a direct bond, where R and R]8 are as defined below;
E5 is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, NR19, S, SO, S02, O, C(O),
Figure imgf000018_0002
or a direct bond, where R,9 and R20 are as defined below;
E6 is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, NR2], S, SO, S0 , O, C(O), or a direct bond, where R2ι is as defined below;
E7 is hydrogen, NR22R23, OR22, SR22, SOR22, S02R22, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocyclyl, aryl, or heteroaryl, where R22 and R23 are as defined below;
R5 is hydrogen or lower alkyl; . where
Z] is heteroarylene;
lower alkylene, lower alkenylene, lower alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, C(0)NR24, NR24C(0), S02NR24, NR24S02, NR24, S, SO, S02,
O, C(O), OC(O), C(0)0, or a direct bond, where R24 is as defined below;
Z3 is alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocyclyl, heteroaryl, aryl, NR25R26,
OR25, or hydrogen, where R25 and R26 are as defined below;
R , R8, R9, Rio, R11, R12, Ri3> Rι4> R15, Ri6, Rπ> Rιs> R19, R2o, R21; R22, 23> R24> R25> R26 are, independently, hydrogen, alkyl, alkynyl, alkenyl, cycloalkyl, aryl, heterocyclyl, or heteroaryl;
or a pharmaceutically acceptable salt, solvate, biohydrolyzable ester, biohydrolyzable amide, affinity reagent, or prodrug thereof.
Compounds of the present invention which are currently preferred for their high biological activity are listed below in Tables 1 A and IB; variables below are with reference to the generic structure (I). Table IA
Figure imgf000020_0001
(I)
Figure imgf000020_0002
Figure imgf000021_0001
Figure imgf000022_0001
Figure imgf000023_0001
Figure imgf000024_0001
Figure imgf000025_0001
Figure imgf000026_0001
Figure imgf000027_0001
Figure imgf000028_0001
Figure imgf000029_0001
Figure imgf000030_0001
Figure imgf000031_0001
Figure imgf000032_0001
Figure imgf000033_0001
Figure imgf000034_0001
Figure imgf000035_0001
Figure imgf000036_0001
Figure imgf000037_0001
Figure imgf000038_0001
Figure imgf000039_0001
Figure imgf000040_0001
Figure imgf000041_0001
Figure imgf000042_0001
Figure imgf000043_0001
Figure imgf000044_0001
Figure imgf000045_0001
Figure imgf000046_0001
Figure imgf000047_0001
Figure imgf000048_0001
Figure imgf000049_0001
Figure imgf000050_0001
Figure imgf000051_0001
Figure imgf000052_0003
Table IB
Figure imgf000052_0001
(I)
Figure imgf000052_0002
Figure imgf000053_0001
Figure imgf000054_0001
Compounds of the present invention which are currently preferred for their high biological activity are listed by name below in Tables 2A and 2B
Table 2A
Figure imgf000054_0002
Figure imgf000055_0001
Figure imgf000056_0001
Figure imgf000057_0001
Figure imgf000058_0001
Figure imgf000059_0001
Figure imgf000060_0001
Figure imgf000061_0001
Figure imgf000062_0001
Figure imgf000063_0001
Figure imgf000064_0001
Table 2B
Figure imgf000064_0002
Figure imgf000065_0001
Preferred embodiments of the invention include compounds of general formula (II) where
Ri is methyl, tπfluoromethyl, ethyl, isopropyl, n-propyl, tert-butyl, 3- methoxycyclopentyl, furan-2-ethynyl, 4-methyl-l -pentyl, 2-thιophenesulfanylmethyl, 4- trifluoromethylcyclohexyl, 3-amιnophenoxymethyl, 3-(4-mo holme)-l -propyl, 2-(3- tetrazolyl)-l -ethyl, 2-(3-pyrιdyl)-l -ethyl, 2-(3-furyl)-l -ethyl, 2-(2-thιazolyl)-l -ethyl, 3,3,3- trιfluoro-1 -propyl, 4,4,4-trιfluoro- 1 -butyl, 2-(4-trιfluorophenyl)-l -ethyl, thιophene-3 -ethynyl, 2-mtrophenoxymethyl, 3-nιtrophenoxymethyl, ethynyl, 2-propynyl, 2-butynyl, phenylethynyl, or vinyl;
R2 is 5-methylthιophene-2-methyl, 2-furanmethyl, thιophene-2-methyl, benzothιophene-2-methyl, benzofuran-2-methyl, 4-fluorobenzyl, 3-phenyl-l -propyl, 3- phenyl-2-methyl-l -propyl, 3-(2-pyrιdyl)-l -propyl, 3-(thιophene-2-yl)-l -propyl, 4-phenyl-l- butyl, 3-phenyl-2-propene-l-yl, 3-(benzofuran-3-yl)-l-propyl, 3-(benzothιophene-3-yl)-l- propyl, 3-(furan-2-yl)-l -propyl, 3 -(2-thιazolyl)-l -propyl, 3-(pyπmιdm-2-yl)-l -propyl, 3- phenyl-2-ethyl-l -propyl, benzyloxymethyl, 2-benzyloxy- 1 -ethyl, 3-(3-pyrιdyl)-l -propyl, 2- phenyl-1 -ethyl, 3-benzyloxy-l -propyl. 4-phenylcyclohexylmethyl, 3-(furan-3-yl)-l-propyl, l,2,3,4-tetrahydronaphthalene-2-methyl, 4-bιphenylpropyl, 3-phenyl-l -butyl, 2,3- dιhydrobenzo[l,4]dιoxιne-2-methyl, 2-naphthylmethyl, chroman-2-methyl, 3-phenyl-2- methyl-2-propene-l-yl, 3-bιphenyl, 4-phenyl-3-methyl-2-butyl, 4-(3-thιophenyl)-2-butyl, benzothιophene-3-methyl, benzoxazole-2-methyl, 4-(3-furyl)-2-butyl, 3-(4-chlorophenyl)-l- propyl, 3-phenoxyphenyl-l -propyl, or benzyl;
R3 is hydrogen, isobutyl, or methyl;
R is tert-butyl, 1-propoxy-l -ethyl, 4-(benzyloxycarbonylamιno)-l -butyl, 2-(2- (benzyloxycarbonylammo)- 1 -ethylsulfanyl)-2-propyl, 4-(2-pyrιdylcarbonylammo)-2-methyl- 2-butyl, 4-ιsobutoxycarbonylamιno-l -butyl, 3-pyπdylmethyl, 3-(2-thιophenecarbonylamιno)- 2-methyl-2-propyl, 4-propoxycarbonylammo-2-butyl, 4-(2-naphthylacetylammo)-l -butyl, 1- ethoxycarbonylammo- 1 -ethyl, 4-(2-pyrιdylcarbonylamιno)-2-methyl-2-butyl, 3- (benzyloxycarbonylamιno)-l -propyl, 1 -methanesulfanyl- 1 -ethyl, 3-(2-pyπdylcarbonylamιno)- 2-methyl-2-propyl, 3-(2-thιophenecarbonylammo)-2-propyl, 3-carbamoylammo-l -propyl, 4- (4-pyrιdylcarbonylammo)-2-methyl-2-butyl, 2-ethoxycarbonylamιno-l -propyl, 4- hydroxybenzyl, 4-chlorobenzyl, l-(tetrahydrofuran-3-yloxy)-l-ethyl, 1 -methanesultenyl- 1 - ethyl, 4-fluorobenzyl, 3-(ιmιno-benzenesulfonylamιno)-methylammo-l -propyl, 4- propoxycarbonylammo-1 -butyl, 3-(ιmιno-(2,3,6-tπmethyl-4- methoxybenzenesulfonylamιno))-methylammo-l -propyl, 2-(2-(l,3,4- thιadιazol)ylammosulfonyl)- 1 -ethyl, 3-methylcarbamoylammo-l -propyl, 4- benzyloxycarbonylammobenzyl, isopropyl, 2-(3-pyrιdylcarbonylammo)-l -ethyl, 1,1- dimethyl- 1 -propyl, 2-(2-thιophene)-l -ethyl, cyclohexyl, 3-phenylcarbamoylammo-l -propyl, 4-cyclopentylacetylammo- 1 -butyl, 4-(3-methoxybenzoylammo)- 1 -butyl, 4- ethoxycarbonylammo- 1 -butyl, 4-ethoxycarbonylamιno- 1 -butyl, 2-(2-(ethoxycarbonylammo)- l-ethylsulfanyl)-2-propyl, 2-butyl, 1-methoxy-l -ethyl, 1-hydroxy-l -ethyl, 2-(methoxy methylammocarbonyl)- 1 -ethyl, 2-(4-ethoxycarbonyl-l-pιperazιnecarbonyl)-l -ethyl, 2- guanιdιnesulfonyl-1 -ethyl, 2-methyl-4-(2-pyrιdylcarbonylamιno)-2-butyl, 2-(methyl benzylammocarbonyl)- 1 -ethyl, 2-(4-morpholmecarbonyl)- 1 -ethyl, 2- pyridylcarbonylammomethyl, acetylaminomethyl, 1 -isobutoxy- 1 -ethyl, carbamoylammomethyl, dimethylammocarbonylmethyl, 2-dιmethylammosulfonyl-l -ethyl, 2- methanesulfanyl-2-propyl, 2-hydroxy-2-propyl, 4-(2-pyrιdylcarbonylammo- 1 -butyl, 2- (dimethylammocarbonyl)- 1 -ethyl, 2-methanesulfonyl- 1 -ethyl, 1 -(2-pyπdylmethoxy)- 1 -ethyl, 1-benzyloxy-l -ethyl, phenyl, 2 -methyl- 1 -propyl, or 2 -phenyl- 1 -ethyl, R5 is hydrogen, methyl, ethyl, or propyl; and
Re is 2-thιazolyl, 2-pyπdyl, 2-pyπmιdιnyl, 2-(l,3,4-thιadιazolyl), 3-(5- methyhsoxazolyl), 3-pyrιdyl, 3-mdolyl, 2-(5-methylthιazolyl), 3-hydroxy-2-pyrιdyl, 2-(5- ethanesulfanyl-l,3,4-thιadιazolyl), 2-benzothιazolyl, 6-methoxy-2-benzothιazolyl, 4-pyrιdyl, 2-pyrazmyl, 3-quιnolιnyl, 2-(5-methyl-l,3,4-thιadιazolyl), 6-ammo-3-pyrιdyl, 6-ammo-2- pyπdyl, 4-ammo-3-pyrιdyl, or 3-ammo-2-pyrιdyl.
Other preferred embodiments of the invention include compounds of general formula (II) where where Ri , R3, R4, R5 and R are as defined above and R2 is 5-phenyl-l -pentyl, 3-(4-
trifluorophenyl)- 1 -propyl, 3-(4-methylphenyl)-l -propyl, 3-(4-phenoxyphenyl)-l -propyl, 3-(4- tert-butylphenyl)-l -propyl, 3 -(4-methoxyphenyl)- 1 -propyl, 3-(4-trιfluorophenyl)-l -propyl, or 3-(3-phenoxyphenyl)-l -propyl;
Particularly preferred embodiments of the invention include compounds of general formula (II) where R, is methyl, ethyl, n-propyl, isopropyl, 4-methyl- 1 -pentyl, 2- thiophenesulfanylmethyl, 3-amιnophenoxymethyl, 2-(3-tetrazolyl)-l -ethyl, 2-(3-pyrιdyl)-l- ethyl, 2-(3-furyl)-l -ethyl, 2-(2-thιazolyl)-l -ethyl, 3,3,3-trιfluoro-l-propyl, 2-(4- trifluorophenyl)- 1 -ethyl, thιophene-3-ethynyl, 2-mtrophenoxymethyl, 3-mtrophenoxymethyl, or vinyl; R2 is 5-methylthιophene-2-methyl, 2-furanmethyl, thιophene-2-methyl, benzothιophene-2-methyl, benzofuran-2-methyl, 4-fluorobenzyl, 3-phenyl-l -propyl, 3- phenyl-2-methyl-l -propyl, 3-(2 -pyπdyl)- 1 -propyl, 3-(thιophene-2-yl)-l -propyl, 4-phenyl-l- butyl, 3-phenyl-2-propene-l-yl, 3-(benzofuran-3-yl)-l -propyl, 3-(benzothιophene-3-yl)-l- propyl, 3-(furan-2-yl)-l-propyl, 3-(2-thιazolyl)-l -propyl, 3-(pyrιmιdιn-2-yl)-l -propyl, 3- phenyl-2-ethyl-l -propyl, benzyloxymefhyl, 2-benzyloxy- 1 -ethyl, 3-(3-pyrιdyl)-l -propyl, 2- phenyl-1 -ethyl, 3-benzyloxy-l -propyl. 4-phenylcyclohexylmethyl, 3-(furan-3-yl)-l -propyl, l,2,3,4-tetrahydronaphthalene-2-methyl, 3-phenyl-l -butyl, 2,3-dιhydrobenzo[l,4]dιoxme-2- methyl, 2-naphthylmethyl, chroman-2-methyl, 3-phenyl-2-methyl-2-propene-l-yl, 4-phenyl- 3-methyl-2-butyl, 4-(3-thιophenyl)-2-butyl, benzothιophene-3-methyl, benzoxazole-2- methyl, 4-(3-furyl)-2-butyl, 3-(4-chlorophenyl)-l -propyl, or benzyl;
R3 is hydrogen;
R4 is tert-butyl, 4-(benzyloxycarbonylammo)- 1 -butyl, 2-(2- (benzyloxycarbonylamino)- 1 -ethylsulfanyl)-2-propyl, 3-pyrιdylmethyl, 4-(2- naphthylacetylamino)- 1 -butyl, 3 -(benzyloxycarbonylamino)- 1 -propyl, 3-carbamoylamιno- 1 - propyl, 4-hydroxybenzyl, 4-chlorobenzyl, 4-fluorobenzyl, 3-(ιmιno-(2,3,6-tτιmethyl-4- methoxybenzenesulfonylammo))-methylamιno- 1 -propyl, 4-benzyloxycarbonylammobenzyl, isopropyl, cyclohexyl, 4-cyclopentylacetylamιno-l -butyl, 4-(3-methoxybenzoylammo)-l- butyl, 4-ethoxycarbonylammo- 1 -butyl, 2-(2-(ethoxycarbonylammo)- 1 -ethylsulfanyl)-2- propyl, 2-butyl, 1-methoxy-l -ethyl, 1-hydroxy-l -ethyl, 2-(methoxy methylaminocarbonyl)- 1 - ethyl, 2-(4-ethoxycarbonyl-l-pιperazmecarbonyl)-l -ethyl, 2-guanιdmesulfonyl- 1 -ethyl, 2- methyl-4-(2-pyπdylcarbonylammo)-2-butyl, 2-(methyl benzylammocarbonyl)-l -ethyl, 2-(4- morpholmecarbonyl)- 1 -ethyl, 2-pyrιdylcarbonylamιnomethyl, acetylaminomethyl, 1 - ιsobutoxy-1 -ethyl, carbamoylammomethyl, dimethylammocarbonylmethyl, 2- dιmethylamιnosulfonyl-1 -ethyl, 2-methanesulfanyl-2-propyl, 2-hydroxy-2-propyl, 4-(2- pyridylcarbonylammo- 1 -butyl, 2-(dιmethylammocarbonyl)- 1 -ethyl, 2-methanesulfonyl- 1 - ethyl, 1 -(2-pyπdylmethoxy)- 1 -ethyl, 1-benzyloxy-l -ethyl, phenyl, 2-methyl-l -propyl, or 2- phenyl-1 -ethyl;
R5 is hydrogen; and Ri is 2-thιazolyl, 2-pyrιdyl, 2-pyrιmιdmyl, 2-(l,3,4-thιadιazolyl), 3-(5- methyhsoxazolyl), 3-pyrιdyl, 2-(5-methylthιazolyl), 3-hydroxy-2-pyrιdyl, 2-(5- ethanesulfanyl-l,3,4-thιadιazolyl), 4-pyrιdyl, 2-pyrazιnyl, 2-(5-methyl-l,3,4-thιadιazolyl), 6- amιno-3-pyrιdyl, 6-ammo-2-pyrιdyl, 4-ammo-3-pyrιdyl, or 3-ammo-2-pyπdyl.
Other particularly preferred embodiments of the invention include compounds of general formula (II) where where Ri , R3, R4, R5 and Rg are as defined above and R2 is 5-phenyl-l-
pentyl, 3-(4-trιfluorophenyl)-l -propyl, 3-(4-methylphenyl)-l -propyl, or 3-(4-phenoxyphenyl)- 1 -propyl;
More particularly preferred embodiments of the invention include compounds of general formula (II) where
Ri is methyl, n-propyl, isopropyl, ethyl, or 3,3,3-trιfluoro-l-propyl; R2 is 5-methylthιophene-2-methyl, 3-phenyl- 1 -propyl, 4-phenyl- 1 -butyl, 2-phenyl- 1 - ethyl, 4-phenylcyclohexylmethyl, 3-(4-chlorophenyl)-l -propyl, or benzyl;
R3 is hydrogen;
R) is tert-butyl, isopropyl, 2-butyl, 1-methoxy-l -ethyl, 1-hydroxy-l -ethyl, 2- methanesulfanyl-2 -propyl, or 2-hydroxy-2-propyl; R5 is hydrogen; and R6 is 2-thιazolyl, 2-pyπdyl, 3-pyrιdyl, or 4-pyrιdyl.;
Other more particularly preferred embodiments of the invention include compounds of general formula (II) where where Rj, R3, R4 and R5 are as defined above ; R2 is 5-phenyl-
1 -pentyl, 3-(4-trιfluorophenyl)-l -propyl, 3-(4-methylphenyl)-l -propyl, or 3-(4- phenoxyphenyl)- 1 -propyl; and R*, is 2-(l,3,4-thιadιazolyl); The compounds of the present invention are inhibitors of matrix metalloproteases, TNF converting enzyme, and TNF activity from whole cells. The compounds of the present invention may also inhibit shedding of pathologically significant cell surface protein ectodomains. The compounds of the present invention may also inhibit CD23 proteolysis The invention described herein is additionally directed to pharmaceutical compositions and methods of inhibiting matrix metalloprotease and/or TNF activity and/or CD23 proteolytic fragment activity in a mammal, which methods comprise administering to a mammal in need of a therapeutically defined amount of a compound of formula (I) or (II), defined above, as a single or polymorphic crystalline form or forms, an amorphous form, a single enantiomer, a racemic mixture, a single stereoisomer, a mixture of stereoisomers, a single diastereoisomer, a mixture of diastereomers, a solvate, a pharmaceutically acceptable salt, a solvate, a prodrug, a biohydrolyzable ester, or a biohydrolyzable amide thereof.
According to a further aspect of the present invention there is provided a compound of formula (II) as defined above or a pharmaceutically acceptable salt, solvate, biohydrolyzable ester, biohydrolyzable amide, affinity reagent or prodrug thereof.
Thus, the present invention provides a method of inhibiting a matrix metalloprotease, comprising the step of administering to a mammal m need thereof a pharmacologically effective amount of a compound of the present invention. The invention further provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a pharmacologically effective amount of a compound of the present invention sufficient to inhibit a matrix metalloprotease. A matrix metalloprotease- inhibiting amount can be an amount that reduces or inhibits a matrix metalloprotease activity in the subject
According to a further aspect of the invention there is provided the use of a compound of the present invention in the preparation of a medicament to inhibit a matrix metalloprotease The present invention further provides a method of inhibiting the lntracellular release of tumor necrosis factor alpha, comprising the step of administering to a mammal in need thereof a pharmacologically effective amount of a compound of the present invention. The invention further provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a pharmacologically effective amount of a compound of the present invention sufficient to inhibit cellular release of mature tumor necrosis factor. An amount sufficient to inhibit cellular release of mature tumor necrosis factor can be an amount that reduces or inhibits cellular release of mature tumor necrosis factor in the subject.
According to a further aspect of the invention there is provided the use of a compound of the present invention m the preparation of a medicament to inhibit the cellular release of mature tumor necrosis factor alpha.
Also provided is a method of inhibition of shedding of cell surface protein ectodomains, comprising the step of administering to a mammal m need thereof a pharmacologically effective amount of a compound of the present invention. The invention further provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a pharmacologically effective amount of a compound of the present invention sufficient to inhibit shedding of cell surface protein ectodomains. An amount sufficient to inhibit shedding of cell surface protein ectodomains can be an amount that reduces or inhibits shedding of one or more cell surface protein ectodomains, such as L-selectin, fibronectin, thyrotropm stimulating hormone receptor, transforming growth factor alpha precursor, low density lipoprotein receptor, beta amyloid precursor protein, mterleukm-6 receptor alpha subunit, Fas ligand, CD40 ligand, epidermal growth factor receptor, macrophage colony stimulating factor, interleukin- 1 receptor type II, CD30, and tumor necrosis factor receptors type I and II, in the subject. According to a further aspect of the invention there is provided the use of a compound of the present invention in the preparation of a medicament to inhibit the shedding of cell surface protein ectodomains.
A method of inhibiting CD23 proteolysis, comprising the step of administering to a mammal in need thereof a pharmacologically effective amount of a compound of the present invention. The invention further provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a pharmacologically effective amount of a compound of the present invention sufficient to inhibit CD23 proteolysis. An amount sufficient to inhibit CD23 proteolysis can be an amount that reduces or inhibits CD23 proteolysis in the subject.
According to a further aspect of the invention there is provided the use of a compound of the present invention m the preparation of a medicament to inhibit CD23 proteolysis.
Additionally provided is a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a pharmacologically effective amount of a compound of the present invention sufficient to decrease, or inhibit, a malignant growth.
According to a further aspect of the invention there is provided the use of a compound of the present invention in the preparation of a medicament to decrease or inhibit a malignant growth.
Further, the present invention provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a pharmacologically effective amount of a compound of the present invention sufficient to treat arthritis. Such an amount can be an amount that relieves, i.e., reduces or eliminates, one or more physiologic characteristic of arthritis. According to a further aspect of the invention there is provided the use of a compound of the present invention in the preparation of a medicament to treat arthritis.
Also provided is a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a pharmacologically effective amount of a compound of the present invention sufficient to treat diabetes. Such an amount can be an amount that reduces or eliminates one or more of the complications associated with diabetes. Additionally, the present invention contemplates treating any of these diseases/conditions m a subject by administering to the subject the recited pharmaceutical composition.
According to a further aspect of the invention there is provided the use of a compound of the present invention m the preparation of a medicament to treat diabetes.
The compounds of the present invention can be administered to any mammal in need of inhibition of matrix metalloprotease activity, CD23 proteolysis, shedding of cell surface protein ectodomains and/or TNF activity. Such mammals can include, for example, horses, cows, sheep, pigs, mice, dogs, cats, primates such as chimpanzees, gorillas, rhesus monkeys, and, most preferably humans.
Certain examples of the invention also are orally bioavailable in animals and possess oral activity in animal models of disease.
Salts encompassed within the term "pharmaceutically acceptable salts" refer to non-toxic salts of the compounds of this invention which are generally prepared by reacting the free base with a suitable organic or inorganic acid or by reacting the acid with a suitable organic or inorganic base. Representative salts include the following salts: Acetate,
Benzenesulfonate, Benzoate, Bicarbonate, Bisulfate, Bitartrate, Borate, Bromide, Calcium Edetate, Camsylate, Carbonate, Chloride, Clavulanate, Citrate, Dihydrochloπde, Edetate,
Edisylate, Estolate, Esylate, Fumarate, Gluceptate, Gluconate, Glutamate, Glycollylarsamlate, Hexylresorcinate, Hydrabamme, Hydrobromide, Hydrocloπde, Hydroxynaphthoate, Iodide, Isethionate, Lactate, Lactobionate, Laurate, Malate, Maleate, Mandelate, Mesylate, Methylbromide, Methylnitrate, Methylsulfate, Monopotassium Maleate, Mucate, Napsylate, Nitrate, N-mefhylglucamine, Oxalate, Pamoate (Embonate), Palmitate, Pantothenate, Phosphate/diphosphate, Polygalacturonate, Potassium, Sahcylate, Sodium, Stearate, Subacetate, Succmate, Tannate, Tartrate, Teoclate, Tosylate, Tπethiodide, Tπmethylammonium and Valerate.
Other salts which are not pharmaceutically acceptable may be useful in the preparation of compounds of formula (I) or (II) and these form a further aspect of the invention.
Also included within the scope of the invention are the individual enantiomers of the compounds represented by formula (I) or (II) above as well as any wholly or partially racemic mixtures thereof. The present invention also covers the individual enantiomers of the compounds represented by formula above as mixtures with diastereoisomer s thereof which one or more of the three stereocenters are inverted.
As used herein, the term "lower" refers to a group having between one and six carbons.
As used herein, the term "alkyl" refers to a straight or branched chain hydrocarbon having from one to ten carbon atoms, optionally substituted with substituents selected from the group consisting of lower alkyl, lower alkoxy, lower alkylsulfanyl, lower alkylsulfenyl, lower alkylsulfonyl, oxo, hydroxy, mercapto, amino optionally substituted by alkyl, carboxy, carbamoyl optionally substituted by alkyl, ammosulfonyl optionally substituted by alkyl, nitro, cyano, halogen, or lower perfluoroalkyl, multiple degrees of substitution being allowed Examples of "alkyl" as used herein include , but are not limited to, n-butyl, n-pentyl, isobutyl, and isopropyl, and the like.
As used herein, the term "alkylene" refers to a straight or branched chain divalent hydrocarbon radical having from one to ten carbon atoms, optionally substituted with substituents selected from the group consisting of lower alkyl, lower alkoxy, lower alkylsulfanyl, lower alkylsulfenyl, lower alkylsulfonyl, oxo, hydroxy, mercapto, amino optionally substituted by alkyl, carboxy, carbamoyl optionally substituted by alkyl, ammosulfonyl optionally substituted by alkyl, nitro, cyano, halogen, or lower perfluoroalkyl, multiple degrees of substitution being allowed. Examples of "alkylene" as used herein include, but are not limited to, methylene, ethylene, and the like.
As used herein, the term "alkenyl" refers to a hydrocarbon radical having from two to ten carbons and at least one carbon - carbon double bond, optionally substituted with substituents selected from the group consisting of lower alkyl, lower alkoxy, lower alkylsulfanyl, lower alkylsulfenyl, lower alkylsulfonyl, oxo, hydroxy, mercapto, ammo optionally substituted by alkyl, carboxy, carbamoyl optionally substituted by alkyl, ammosulfonyl optionally substituted by alkyl, nitro, cyano, halogen, or lower perfluoroalkyl, multiple degrees of substitution being allowed.
As used herein, the term "alkenylene" refers to an straight or branched chain divalent hydrocarbon radical having from two to ten carbon atoms and one or more carbon - carbon double bonds, optionally substituted with substituents selected from the group consisting of lower alkyl, lower alkoxy, lower alkylsulfanyl, lower alkylsulfenyl, lower alkylsulfonyl, oxo, hydroxy, mercapto, amino optionally substituted by alkyl, carboxy, carbamoyl optionally substituted by alkyl, ammosulfonyl optionally substituted by alkyl, nitro, cyano, halogen, or lower perfluoroalkyl, multiple degrees of substitution being allowed. Examples of "alkenylene" as used herein include, but are not limited to, ethene-l,2-dιyl, propene-l,3-dιyl, methylene- 1,1-dιyl, and the like.
As used herein, the term "alkynyl" refers to a hydrocarbon radical having from two to ten carbons and at least one carbon - carbon triple bond, optionally substituted with substituents selected from the group consisting of lower alkyl, lower alkoxy, lower alkylsulfanyl, lower alkylsulfenyl, lower alkylsulfonyl, oxo, hydroxy, mercapto, ammo optionally substituted by alkyl, carboxy, carbamoyl optionally substituted by alkyl, ammosulfonyl optionally substituted by alkyl, nitro, cyano, halogen, or lower perfluoroalkyl, multiple degrees of substitution being allowed.
As used herein, the term "alkynylene" refers to a straight or branched chain divalent hydrocarbon radical having from two to ten carbon atoms and one or more carbon - carbon triple bonds, optionally substituted with substituents selected from the group consisting of lower alkyl, lower alkoxy, lower alkylsulfanyl, lower alkylsulfenyl, lower alkylsulfonyl, oxo, hydroxy, mercapto, ammo optionally substituted by alkyl, carboxy, carbamoyl optionally substituted by alkyl, ammosulfonyl optionally substituted by alkyl, nitro, cyano, halogen, or lower perfluoroalkyl, multiple degrees of substitution being allowed. Examples of "alkynylene" as used herein include, but are not limited to, ethyne-l,2-dιyl, propyne-l,3-dιyl, and the like.
As used herein, "cycloalkyl" refers to a ahcyclic hydrocarbon group with one or more degrees of unsaturation, having from three to twelve carton atoms, optionally substituted with substituents selected from the group consisting of lower alkyl, lower alkoxy, lower alkylsulfanyl, lower alkylsulfenyl, lower alkylsulfonyl, oxo, hydroxy, mercapto, ammo optionally substituted by alkyl, carboxy, carbamoyl optionally substituted by alkyl, ammosulfonyl optionally substituted by alkyl, nitro, cyano, halogen, or lower perfluoroalkyl, multiple degrees of substitution being allowed. "Cycloalkyl" includes by way of example cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, or cyclooctyl, and the like.
As used herein, the term "cycloalkylene" refers to an non-aromatic ahcyclic divalent hydrocarbon radical having from three to twelve carbon atoms, optionally substituted with substituents selected from the group consisting of lower alkyl, lower alkoxy, lower alkylsulfanyl, lower alkylsulfenyl, lower alkylsulfonyl, oxo, hydroxy, mercapto, ammo optionally substituted by alkyl, carboxy, carbamoyl optionally substituted by alkyl, ammosulfonyl optionally substituted by alkyl, nitro, cyano, halogen, or lower perfluoroalkyl, multiple degrees of substitution being allowed. Examples of "cycloalkylene" as used herein include, but are not limited to, cyclopropyl- 1,1-dιyl, cyclopropyl- 1,2-dιyl, cyclobutyl- 1,2- diyl, cyclopentyl- 1,3-dιyl, cyclohexyl- 1,4-dιyl, cycloheptyl- 1,4-dιyl, or cyclooctyl- 1,5-dιyl, and the like.
As used herein, the term "cycloalkenyl" refers to a substituted ahcyclic hydrocarbon radical having from three to twelve carbon atoms and at least one carbon-carbon double bond in the ring system, optionally substituted with substituents selected from the group consisting of lower alkyl, lower alkoxy, lower alkylsulfanyl, lower alkylsulfenyl, lower alkylsulfonyl, oxo, hydroxy, mercapto, amino optionally substituted by alkyl, carboxy, carbamoyl optionally substituted by alkyl, ammosulfonyl optionally substituted by alkyl, nitro, cyano, halogen, or lower perfluoroalkyl, multiple degrees of substitution being allowed. Examples of "cycloalkenylene" as used herein include, but are not limited to, 1 - cyclopentene-3-yl, l-cyclohexene-3-yl, l-cycloheptene-4-yl, and the like.
As used herein, the term "cycloalkenylene" refers to a substituted ahcyclic divalent hydrocarbon radical having from three to twelve carbon atoms and at least one carbon-carbon double bond in the ring system, optionally substituted with substituents selected from the group consisting of lower alkyl, lower alkoxy, lower alkylsulfanyl, lower alkylsulfenyl, lower alkylsulfonyl, oxo, hydroxy, mercapto, ammo optionally substituted by alkyl, carboxy, carbamoyl optionally substituted by alkyl, ammosulfonyl optionally substituted by alkyl, nitro, cyano, halogen, or lower perfluoroalkyl, multiple degrees of substitution being allowed Examples of "cycloalkenylene" as used herein include, but are not limited to, 4,5-cyclopentene-l,3-dιyl, 3,4-cyclohexene-l,l-dιyl, and the like
As used herein, the term "heterocyclic" or the term "heterocyclyl" refers to a three to twelve-membered heterocyclic ring having one or more degrees of unsaturation containing one or more heteroatomic substitutions selected from S, SO, S02, O, or N, optionally substituted with substituents selected from the group consisting of lower alkyl, lower alkoxy, lower alkylsulfanyl, lower alkylsulfenyl, lower alkylsulfonyl, oxo, hydroxy, mercapto, ammo optionally substituted by alkyl, carboxy, carbamoyl optionally substituted by alkyl, ammosulfonyl optionally substituted by alkyl, nitro, cyano, halogen, or lower perfluoroalkyl, multiple degrees of substitution being allowed Such a ring may be optionally fused to one or more of another "heterocyclic" πng(s) or cycloalkyl πng(s) Examples of "heterocyclic" include, but are not limited to, tetrahydrofuran, pyran, 1,4-dιoxane, 1,3- dioxane, pipeπdme, pyrrohdme, morphohne, tetrahydrothiopyran, tetrahydrothiophene, and
As used herein, the term "heterocyclylene" refers to a three to twelve- membered heterocyclic ring diradical having one or more degrees of unsaturation containing one or more heteroatoms selected from S, SO, S02, O, or N, optionally substituted with substituents selected from the group consisting of lower alkyl, lower alkoxy, lower alkylsulfanyl, lower alkylsulfenyl, lower alkylsulfonyl, oxo, hydroxy, mercapto, ammo optionally substituted by alkyl, carboxy, carbamoyl optionally substituted by alkyl, ammosulfonyl optionally substituted by alkyl, nitro, cyano, halogen, or lower perfluoroalkyl, multiple degrees of substitution being allowed Such a ring may be optionally fused to one or more benzene πngs or to one or more of another "heterocyclic" rings or cycloalkyl πngs Examples of "heterocyclylene" include, but are not limited to, tetrahydrofuran-2,5-dιyl, morpholme-2,3-dιyl, pyran-2,4-dιyl, l ,4-dιoxane-2,3-dιyl, l,3-dιoxane-2,4-dιyl, prpeπdme- 2,4-dιyl, pιpeπdιne-l ,4-dιyl, pyrrolιdιne-l,3-dιyl, morpholιne-2,4-dιyl, and the like
As used herein, the term "aryl" refers to a benzene ring or to an optionally substituted benzene ring system fused to one or more optionally substituted benzene πngs, optionally substituted with substituents selected from the group consisting of lower alkyl, lower alkoxy, lower alkylsulfanyl, lower alkylsulfenyl, lower alkylsulfonyl, oxo, hydroxy, mercapto, amino optionally substituted by alkyl, carboxy, tetrazolyl, carbamoyl optionally substituted by alkyl, ammosulfonyl optionally substituted by alkyl, acyl, aroyl, heteroaroyl, acyloxy, aroyloxy, heteroaroyloxy, alkoxycarbonyl, nitro, cyano, halogen, or lower perfluoroalkyl, multiple degrees of substitution being allowed. Examples of aryl include, but are not limited to, phenyl, 2-naphthyl, 1-naphthyl, and the like.
As used herein, the term "arylene" refers to a benzene ring diradical or to a benzene πng system diradical fused to one or more optionally substituted benzene πngs, optionally substituted with substituents selected from the group consisting of lower alkyl, lower alkoxy, lower alkylsulfanyl, lower alkylsulfenyl, lower alkylsulfonyl, oxo, hydroxy, mercapto, ammo optionally substituted by alkyl, carboxy, tetrazolyl, carbamoyl optionally substituted by alkyl, ammosulfonyl optionally substituted by alkyl, acyl, aroyl, heteroaroyl, acyloxy, aroyloxy, heteroaroyloxy, alkoxycarbonyl, nitro, cyano, halogen, or lower perfluoroalkyl, multiple degrees of substitution being allowed. Examples of "arylene" include, but are not limited to, benzene- 1,4-dιyl, naphthalene- 1,8-dιyl, anthracene- 1 ,4-dιyl, and the like.
As used herein, the term "heteroaryl" refers to a five - to seven - membered aromatic ring, or to a polycyclic heterocyclic aromatic πng, containing one or more nitrogen, oxygen, or sulfur heteroatoms, where N-oxides and sulfur monoxides and sulfur dioxides are permissible heteroaromatic substitutions, optionally substituted with substituents selected from the group consisting of lower alkyl, lower alkoxy, lower alkylsulfanyl, lower alkylsulfenyl, lower alkylsulfonyl, oxo, hydroxy, mercapto, amino optionally substituted by alkyl, carboxy, tetrazolyl, carbamoyl optionally substituted by alkyl, ammosulfonyl optionally substituted by alkyl, acyl, aroyl, heteroaroyl, acyloxy, aroyloxy, heteroaroyloxy, alkoxycarbonyl, nitro, cyano, halogen, or lower perfluoroalkyl, multiple degrees of substitution being allowed For polycyclic aromatic πng systems, one or more of the rings may contain one or more heteroatoms Examples of "heteroaryl" used herein are furan, thiophene, pyrrole, imidazole, pyrazole, tπazole, tetrazole, thiazole, oxazole, isoxazole, oxadiazole, thiadiazole, isothiazole, pyπdine, pyridazme, pyrazme, pyπmidine, quinoline, lsoquinolme, benzofuran, benzothiophene, mdole, and mdazole, and the like
As used herein, the term "heteroarylene" refers to a five - to seven - membered aromatic ring diradical, or to a polycyclic heterocyclic aromatic ring diradical, containing one or more nitrogen, oxygen, or sulfur heteroatoms, where N-oxides and sulfur monoxides and sulfur dioxides are permissible heteroaromatic substitutions, optionally substituted with substituents selected from the group consisting of lower alkyl, lower alkoxy, lower alkylsulfanyl, lower alkylsulfenyl, lower alkylsulfonyl, oxo, hydroxy, mercapto, ammo optionally substituted by alkyl, carboxy, tetrazolyl, carbamoyl optionally substituted by alkyl, ammosulfonyl optionally substituted by alkyl, acyl, aroyl, heteroaroyl, acyloxy, aroyloxy, heteroaroyloxy, alkoxycarbonyl, nitro, cyano, halogen, or lower perfluoroalkyl, multiple degrees of substitution being allowed For polycyclic aromatic ring system diradicals, one or more of the rings may contain one or more heteroatoms Examples of "heteroarylene" used herein are furan-2,5-dιyl, thιophene-2,4-dιyl, l,3,4-oxadιazole-2,5-dιyl, l,3,4-thιadιazole-2,5- diyl, l,3-thιazole-2,4-dιyl, l,3-thιazole-2,5-dιyl, pyπdιne-2,4-dιyl, pyπdιne-2.3-dιyl, pyπdιne-2,5-dιyl, pyπmιdιne-2,4-dιyl, quιnohne-2,3-dιyl, and the like As used herein, the term "direct bond", where part of a structural vaπable specification, refers to the direct joining of the substituents flanking (preceding and succeeding) the variable taken as a "direct bond". Where two or more consecutive vaπables are specified each as a "direct bond", those substituents flanking (preceding and succeeding) those two or more consecutive specified "direct bonds" are directly joined.
As used herein, the term "alkoxy" refers to the group RaO-, where Ra is alkyl.
As used herein, the term "alkenyloxy" refers to the group RaO-, where Ra is alkenyl.
As used herein, the term "alkynyloxy" refers to the group RaO-, where Ra is alkynyl.
As used herein, the term "aryloxy" refers to the group RaO-, where Ra is aryl.
As used herein, the term "heteroaryloxy" refers to the group RaO-, where Ra is heteroaryl.
As used herein, the term "alkylsulfanyl" refers to the group RaS-, where Ra is alkyl.
As used herein, the term "alkenylsulfanyl" refers to the group RaS-, where Ra is alkenyl.
As used herein, the term "alkynylsulfanyl" refers to the group RaS-, where Ra is alkynyl. As used herein, the term "alkylsulfenyl" refers to the group RaS(0)-, where Ra is alkyl.
As used herein, the term "alkenylsulfenyl" refers to the group RaS(0)-, where Ra is alkenyl.
As used herein, the term "alkynylsulfenyl" refers to the group RaS(0)-, where Ra is alkynyl.
As used herein, the term "alkylsulfonyl" refers to the group RaS02-, where Ra is alkyl.
As used herein, the term "alkenylsulfonyl" refers to the group RaS0 -, where Ra is alkenyl.
As used herein, the term "alkynylsulfonyl" refers to the group RaS02-, where Ra is alkynyl.
As used herein, the term "acyl" refers to the group RaC(0)- , where Ra is alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, or heterocyclyl.
As used herein, the term "aroyl" refers to the group RaC(0)- , where Ra is aryl.
As used herein, the term "heteroaroyl" refers to the group RaC(0)- , where Ra is heteroaryl. As used herein, the term "alkoxycarbonyl" refers to the group RaOC(0)-, where Ra is alkyl.
As used herein, the term "acyloxy" refers to the group RaC(0)0- , where Ra is alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, or heterocyclyl.
As used herein, the term "aroyloxy" refers to the group RaC(0)0- , where Ra is aryl.
As used herein, the term "heteroaroyloxy" refers to the group RaC(0)0- , where Ra is heteroaryl.
As used herein, the term "optionally" means that the subsequently described event(s) may or may not occur, and includes both event(s) which occur and events that do not occur.
As used herein, the term "substituted" refers to substitution with the named substituent or substituents, multiple degrees of substitution being allowed unless otherwise stated.
As used herein, the terms "contain" or "containing" can refer to in-line substitutions at any position along the above-defined alkyl, alkenyl, alkynyl or cycloalkyl substituents with one or more of any of O, S, SO, S02, N, or N-alkyl, including, for example, -CH2-0-CH2-, -CH2-S02-CH2-, -CH2-NH-CH3 and so forth.
As used herein, the term "solvate" is a complex of variable stoichiometry formed by a solute ( this invention, a compound of formula (I) or (II)) and a solvent. Such solvents for the purpose of the invention may not interfere with the biological activity of the solute. Solvents may be, by way of example, water, ethanol, or acetic acid.
As used herein, the term "biohydrolyzable ester" is an ester of a drug substance (in this invention, a compound of general formula (I) or (II) which either a) does not interfere with the biological activity of the parent substance but confers on that substance advantageous properties in vivo such as duration of action, onset of action, and the like, or b) is biologically inactive but is readily converted in vivo by the subject to the biologically active principle. The advantage is that, for example, the biohydrolyzable ester is orally absorbed from the gut and is transformed to (I) or (II) in plasma. Many examples of such are known in the art and include by way of example lower alkyl esters, lower acyloxy-alkyl esters, lower alkoxyacyloxyalkyl esters, alkoxyacyloxy esters, alkyl acylammo alkyl esters, and cholme esters. An example of such a biohydrolyzable ester applied to the general formula (II) is illustrated below in general formula (III).
Figure imgf000084_0001
(ill)
As used herein, the term "biohydrolyzable amide" is an amide of a drug substance (in this invention, a compound of general formula (I) or (II) which either a) does not interfere with the biological activity of the parent substance but confers on that substance advantageous properties m vivo such as duration of action, onset of action, and the like, or b) is biologically inactive but is readily converted in vivo by the subject to the biologically active principle. The advantage is that, for example, the biohydrolyzable amide is orally absorbed from the gut and is transformed to (I) or (II) in plasma. Many examples of such are known m the art and include by way of example lower alkyl amides, α-amino acid amides, alkoxyacyl amides, and alkylaminoalkylcarbonyl amides
As used herein, the term "prodrug" includes biohydrolyzable amides and biohydrolyzable esters and also encompasses a) compounds in which the biohydrolyzable functionality in such a prodrug is encompassed in the compound of formula (I) or (II): for example, the lactam formed by a carboxylic group in R2 and an amme in R4, and b) compounds which may be oxidized or reduced biologically at a given functional group to yield drug substances of formula (I) or (II) Examples of these functional groups are, but are not limited to, 1 ,4-dιhydropyπdιne, N-alkylcarbonyl-l,4-dιhydropyπdme, 1,4- cyclohexadiene, tert-butyl, and the like.
As used herein, the term "affinity reagent" is a group attached to the compound of formula (I) or (II) which does not affect its in vitro biological activity, allowing the compound to bind to a target, yet such a group binds strongly to a third component allowing a) characterization of the target as to localization within a cell or other organism component, perhaps by visualization by fluorescence or radiography, or b) facile separation of the target from an unknown mixture of targets, whether proteinaceous or not protemaceous An example of an affinity reagent according to b) would be biotin either directly attached to (I) or (II) or linked with a spacer of one to 50 atoms selected from the group consisting of C, H, O, N, S, or P in any combination. An example of an affinity reagent according to a) above would be fluorescein, either directly attached to (I) or (II) or linked with a spacer of one to 50 atoms selected from the group consisting of C, H, O, N, S, or P m any combination
The term "pharmacologically effective amount" shall mean 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 by a researcher or clinician. This amount can be a therapeutically effective amount Whenever the terms "alkyl" or "aryl" or either of their prefix roots appear in a name of a substituent (e.g. arylalkoxyaryloxy) they shall be interpreted as including those limitations given above for "alkyl" and "aryl". Alkyl or cycloalkyl substituents shall be recognized as being functionally equivalent to those having one or more degrees of unsaturation. Designated numbers of carbon atoms (e.g Cι_ιo) shall refer independently to the number of carbon atoms in an alkyl, alkenyl or alkynyl or cyclic alkyl moiety or to the alkyl portion of a larger substituent in which the term "alkyl" appears as its prefix root
As used herein, the term "oxo" shall refer to the substituent =0.
As used herein, the term "halogen" or "halo" shall include iodine, bromine, chlorine and fluorine.
As used herein, the term "mercapto" shall refer to the substituent -SH.
As used herein, the term "carboxy" shall refer to the substituent -COOH.
As used herein, the term "cyano" shall refer to the substituent -CN.
As used herein, the term "ammosulfonyl" shall refer to the substituent
S02NH2
As used herein, the term "carbamoyl" shall refer to the substituent -C(0)NH2
As used herein, the term "sulfanyl" shall refer to the substituent -S-
As used herein, the term "sulfenyl" shall refer to the substituent -S(O)- As used herein, the term "sulfonyl" shall refer to the substituent -S(0)2-
The compounds of formula (I) and (II) can be prepared readily according to the following reaction Schemes (in which all variables are as defined before) and Examples or modifications thereof using readily available starting materials, reagents and conventional synthesis procedures. In these reactions, it is also possible to make use of variants which are themselves known to those of ordinary skill in this art, but are not mentioned in greater detail.
The most preferred compounds of the invention are any or all of those specifically set forth m these examples. These compounds are not, however, to be construed as forming the only genus that is considered as the invention, and any combination of the compounds or their moieties may itself form a genus. The following examples further illustrate details for the preparation of the compounds of the present invention. Those skilled in the art will readily understand that known variations of the conditions and processes of the following preparative procedures can be used to prepare these compounds. All temperatures are degrees Celsius unless noted otherwise.
Abbreviations used m the Examples are as follows g = grams mg = milligrams
L = liters mL = millihters psi = pounds per square inch
M = molar
N = normal mM = milhmolar l.V. = intravenous p.o. = per oral s.c. = subcutaneous
Hz = hertz mol = moles mmol = milhmoles mbar = millibar rt = room temperature mm = minutes h = hours mp = melting point TLC = thin layer chromatography
Rf = relative TLC mobility
MS = mass spectrometry NMR = nuclear magnetic resonance spectroscopy APCI = atmospheric pressure chemical lonization ESI = electrospray lonization m/z = mass to charge ratio tr = retention time ether = diethyl ether MeOH = methanol EtOAc = ethyl acetate TEA = tπethylamine DIEA = dnsopropylethylamine BOP = ( 1 -benzotπazolyloxy)tπs(dιmethylamιno)phosphonιum hexafluorophosphate
THF = tetrahydrofuran
DMF = N, N-dimethylformamidε
DBU =l,8-dιazabιcyclo[5.4.0]undec-7-ene DMSO = dimethylsulfoxide
LAH = lithium aluminum hydπde
TFA = tπfluoroacetic acid
EDC =l-ethyl-3-(3-dιmethylammopropyl)-carbodnmιde hydrochloπde HOBt = 1 -hydroxybenzotπazole
LDA = lithium dnsopropylamide
THP = tetrahydropyranyl
NMM = N-methylmorphohne, 4-mefhylmorpholme
HMPA = hexamethylphosphoπc tπamide DMPU = 1 ,3-dιmethypropylene urea d = days mm = minutes ppm = parts per million kD = kiloDalton LPS = lipopolysacchaπde
PMA = phorbol myπ state acetate
SPA = scintillation proximity assay
EDTA = efhylenediamine tetraacetic acid
FBS = fetal bovme serum PBS = phosphate buffered saline solution
ELISA = enzyme - linked immunosorbent assay
Several of the following examples represent pairs of stereoisomers which were separated as diastereoisomers but were not identified therein. Determination and/or preparation of the R and S isomers can advantageously be approached by stereoselective chemical methods, see "Advanced Organic Chemistry", Carey and Sundberg, 3rd edition.
Plenum Press, 1990, 596, by analytical methods such as X-ray crystallography, or by determination of biological activity and subsequent correlation to biologically active compounds of known stereochemistry.
GENERAL REACTION SCHEMES
Compounds of the invention may be prepared by methods known in the art, where such a method is shown m reaction Scheme 1.
Reaction Scheme 1
Figure imgf000090_0001
IV V
Figure imgf000090_0002
Vll " Rj, R2, R3, R,, R5, and R are as defined as for formula (II).
RPG] is selected from the group consisting of benzyl or 2-tetrahydropyranyl. R27 is chosen from the group consisting of hydroxyl, 0-C6F5, or halogen.
When R27 is hydroxyl, the conversion of (V) and (VI) to (VII) involves methods known in peptide chemistry; for example, the reaction may be conducted using HOBt in combination with a dehydrating agent such as dicyclohexylcarbodiimide in a suitable solvent, such as DMF. When R27 is 0-C6F5, the conversion of (IV) to (V) is conducted by treating (IV) m a suitable solvent such as dichloromethane with pentafluorophenyl tπfluoroacetate in the presence of pyπdine, or with EDC and pentafluorophenol in a suitable solvent such as dichloromethane. The displacement reaction to produce (VII) employing (VI) is carried out in the presence of a suitable solvent such as dioxane, THF, dichloromethane, or DMF, at a temperature of 0 °C to 140 °C. The reaction is effected in the presence of an organic base such as NMM or TEA. It is understood that if Ri in (VII) contains a monosubstituted alkenyl group, then addition of an aryl or heteroaryl bromide to (VII) m the presence of tetrakιs(tπphenylphosphme)palladιum and TEA m DMF or THF affords the alkenylaryl or alkynylheteroaryl intermediate. If R] in (VII) contains a terminal alkynyl group, then addition of an aryl or heteroaryl bromide to (VII) in the presence of tetrakιs(tπphenylphosphme)palladιum, Cul, and TEA in DMF or THF affords the alkynylaryl or alkynylheteroaryl intermediate. These Ri substituents may be reduced with palladium on carbon and hydrogen to the saturated species. The removal of the RPG, group where RPG] is benzyl may be achieved by hydrogenation of (VII) with palladium on barium sulfate in a suitable solvent such as ethanol or THF, or, where RPG] is 2-tetrahydropyranyl, by hydrolysis with aqueous acetic acid at a temperature of 20 °C to 100 °C.
Reaction Scheme 2 depicts the synthesis of a compound of formula (IV).
Reaction Scherre 2
Figure imgf000092_0001
VIII IX
Figure imgf000092_0002
Figure imgf000092_0003
XII IV
R, and R2 are as defined for formula (II).
R280 is a nucleofugal group such as methanesulfonate or p-toluenesulfonate.
RPG] is defined as for reaction scheme 1. The acid of formula (VIII) may be converted to the alcohol of formula (IX) by treatment with HOBt, O-benzylhydroxylamme hydrochloπde or 2- tetrahydropyranyloxyamine, NMM, and a carbodnmide reagent such as EDC in a suitable solvent such as DMF. The alcohol of formula (IX) may be converted to (X) by treatment with methanesulfonyl chloride and pyπdme in a suitable solvent such as dichloromethane. The conversion of (X) to (XI) may be conducted by treatment with potassium carbonate in a suitable solvent such as acetone or 2-butanone, at temperature of 20 °C to 90 °C. Alternatively, (IX) may be converted directly to (XI) by treatment with tπphenylphosphine and diethyl azodicarboxylate or another azodicarbonyl diester or diamide in a suitable solvent such as THF at a temperature of -78 °C to 50 °C. (XI) where R, contains a primary or secondary hydroxyl group may be treated with a thiol, phenol, or heteroaryl species containing an NH group m the presence of tπphenylphosphine and diethyl azodicarboxylate or another azodicarbonyl diester or diamide in a suitable solvent such as THF to afford the respective product of hydroxy displacement in (XI). The compound of formula (XI) may be converted to (XII) by treatment with an inorganic base such as sodium hydroxide in water or water in combination with a water - soluble organic cosolvent such as methanol or THF, followed by acidification with an acidic solution such as aqueous citric acid or aqueous sodium bisulfate. The compound of formula (XII) may be converted to (IV) by treatment with acetic anhydride and formic acid or by treatment with formic acetic anhydride in pyπdme in the presence or absence of a suitable cosolvent such as dichloromethane. An alternative route of preparation of compounds of formula (IX) is depicted in reaction Scheme 3.
Reaction Scheme 3
Figure imgf000093_0001
VIII
Figure imgf000093_0002
IX
R] and R2 are as defined as for formula (II). RPG, is defined as for reaction scheme 1.
R29 is chosen from the group consisting of lower alkoxy or oxazolιdιnon-1-yl, where the 4 and 5 positions of an oxazolιdιnon-1-yl group may be substituted with a lower alkyl, aryl, or lower alkylaryl group and where such an oxazohdinon- 1 -yl substituent may exist as a single stereoisomer or as a mixture of stereoisomers.
A carbonyl compound of formula (XIII), where R29 is an alkoxy group such as methoxy or tert-butoxy, may be treated with a strong base such as LDA m a solvent such as THF at a temperature of from -78 °C to 0 °C, followed by treatment with the aldehyde (XIV) to provide (XV). Where R29 is a oxazohdinon- 1-yl substituent, treatment of (XIII) with a Lewis acid such as dι(n-butyl)boron tπfluoromethanesulfonate in the presence of DIEA in a suitable solvent such as dichloromethane at a temperature of 0 °C, followed by addition of the aldehyde (XIV) provides (XV) Treatment of (XV) with aqueous base in the presence or absence of hydrogen peroxide affords (VIII) upon acidification. The acid (VIII) may be converted directly to (IX) as in reaction Scheme 2, or may be treated with a dehydrating agent such a p-toluenesulfonyl chloride m pyπdine or with tπphenylphosphine and diethyl azodicarboxylate m a suitable solvent such as THF, to afford the lactone (XVI). Treatment of the lactone (XVI) with H2NO-RPGι in the presence of a Lewis acid such as tπmethylalummum in a suitable solvent such as toluene affords the alcohol (LX).
A preparation of compounds of general formula (IV) is depicted m reaction Scheme 4.
Reaction Scheme 4
Figure imgf000095_0001
Ri and R2 are as defined as for formula (II). R30, R3 ! , R32, and R33 may be, independently, alkyl, alkenyl, alkynyl, or hydrogen. R34 and R35 are selected from the group consisting of aryl, heteroaryl, alkynyl, or alkenyl, with the proviso that the unsaturated carbons of alkenyl and alkynyl groups are directly bonded to Lj
L] is selected from the group consisting of bromide, iodide, or tπfluoromethanesulfonate. L2 is tπ(lower alkyl)stannyl or -B(OH)2. RPG] is defined as for reaction scheme 1.
The lactam of general formula (XVII) may be treated with a palladium catalyst such as tetrakιs(tπphenylphosphιne)palladιum and R34-L] m a solvent such as acetonitπle in the presence of a tertiary amine base such as NMM at a temperature of from 20 °C to 200 °C to afford (XVIII). Reduction of the olefinic group in (XVIII) with hydrogen and a metal catalyst such as palladium on carbon and conversion of the lactam (XI) to the acid (IV) proceeds as outlined in reaction Scheme 2. Alternately, the olefin in compounds of general formula (XVIII) may be left in place and manipulation of the lactam (XVIII) is carried out as described in reaction Scheme 2 to afford (XIX). (XIX) may be converted to (IV) as descπbed in reaction Scheme 2 with or without reduction of the olefin in (XIX), as appropπate. The alkyne (XX) may be treated with tπ(butyltm) hydride in the presence of a radical initiator such as azobιs(ιsobutyronιtπle) to afford an alkyl tin intermediate (XXI), which may be freated with R34-L] and a catalyst such as Pd(PPh3)4 m a solvent such as DMF in the presence or LiCl to afford (XXII). Alternately, (XXI) may be treated with iodine in an organic solvent such as ether to afford the destannylated vmyl iodide. The iodide may be treated with R34-L2 in the presence of a palladium catalyst such as Pd(PPh3)4 in a solvent such as DMF or THF at a temperature of 25 °C to 140 °C to afford (XXII). The alkyne (XXIII ) may be treated with R35-L1, CuCl or Cul and an alkylamme such as TEA in the presence of a catalyst such as Pd(PPh3)4 to afford (XXIV). (XXIV and (XXII) may be manipulated to the intermediate (IV) via the operations described in reaction scheme 2
Reaction Scheme 5 depicts the preparation of compounds of general formula (VIII) Fraction Scherre 5
Figure imgf000097_0001
XXXJ XXXIII
Figure imgf000097_0002
XXXIV VIII
Ri and R2 are as defined for formula (II). R36 is lower alkyl.
RPGi is as defined for reaction Scheme 1. L3 is bromide, iodide, or tπfluoromethanesulfonyloxy
R30, R ι, and R32 are as defined for reaction scheme 4
R37 and R38, may be, independently, alkyl, alkenyl, alkynyl, or hydrogen.
The ketoester of general formula (XXVII), if not commercially available, may be prepared by reaction of ester (XXV) with a strong base such as LDA followed by treatment with the aldehyde (XIV). The resulting hydroxyester (XXVI) may be used directly or converted to the ketoester (XXVII) by oxidation with, for example, pyπdmium dichromate in a solvent such as dichloromethane Alternately the acid chloride (XXVIII) may be condensed with 2,2-dιmethyl-4,6-dιoxo-l,3-dιoxane in the presence of pyπdine to afford (XXIX), which may be freated with excess R35-OH at a temterature of from 25 °C to 150 °C to provide
(XXVII). The ketoester of general formula (XXVII) may be reduced with a reducing agent such as sodium borohydπde to afford the hydroxyester (XXVI). Alternately, a chiral catalyst or chiral ligand in the presence of a reducing agent such as hydrogen or a metal hydride such as borane or LAH may be employed to afford (XXVI) with chiral induction at the newly formed asymmetric center. The alcohol (XXVI) may be converted to (XXX) by treatment with a strong base such as LDA in a suitable solvent such as THF, followed by the addition of R2-L3 in the presence or absence of a cosolvent such as DMPU. Where R2 contains an unsaturated group, (XXX) may be reduced under appropriate conditions. Removal of the ester group by hydrolysis with aqueous hydroxide or, in the case where R36 is tert-butyl, by treatment with a strong acid such as TFA, affords (VIII) Hydroxy acid (XXXI) is obtained by hydrolysis of the ester group of (XXVI) with aqueous alkali. (XXXI) may be obtained by freatment of (XXVI) with TFA, where R36 is tert-butyl Coupling of the hydroxy acid (XXXI) with an allylic alcohol (XXXII) m the presence of a dehydrating agent such as EDC and a catalyst such as 4-dιmethylammopyπdme provides the ester (XXXIII). Alternately, protection of the alcohol functionality of ester (XXVI) with, for example, a tert- butyldimefhylsilyl group may be required before processing of (XXVI) to the acid.
Hydrolysis of the ester grouping of the protected (XXVI) as before with aqueous base followed by activation of the acid functionality as its acid chloride with oxalyl chloride and addition of the alcohol (XXXII) m the presence of an organic base such as TEA provides the ester (XXXIII) with the hydroxyl group protected. Deprotection of the hydroxyl group, if so protected, and treatment of the resulting ester (XXXIII) with a sfrong base such as LDA in a solvent such as 1 ,2-dιmethoxyethane at a temperature of -78 °C, followed by warming of the mixture to a temperature of between 0 °C and 90 °C, followed by acidification of the mixture provides the acid (XXXIV). Reduction of the olefmic group in (XXXIV) with hydrogen and a metal catalyst such as palladium on carbon provides the acid (VIII). Alternately, the olefin in compounds of general formula (XXXIV) may be left in place until a later stage and then saturated with, for example, hydrogen gas in the presence of palladium on carbon.
The preparation of compounds of general formula (VI) is shown in reaction Scheme
6.
Reaction Scheme 6
Figure imgf000099_0001
XXXVIII VI
R3, R4, R5, and R6 are as defined for general formula (II).
RPG2 is selected from the group consisting of tert-butoxycarbonyl, allyloxycarbonyl, or benzyloxycarbonyl .
R39 is selected from the group consisting of 1 -benzotπazolyloxy, or bromine. The acid of formula (XXXV) may be converted in situ to (XXXVI), where R39 is bromine, by treatment with bromo-tπs(pyrrolιdιno)phosphonιum hexafluorophosphate m a suitable solvent such as DMF in the presence of an organic base such as DIEA. The acid of formula (XXXV) may be converted in situ to (XXXVI), where R39 is benzotπazolyloxy, by treatment with BOP in a suitable solvent such as DMF in the presence of an organic base such as NMM. Addition of the amine (XXXVII) in the displacement step in the presence of a suitable solvent such as DMF and an organic base such as DIEA affords the amide (XXXVIII) Alternatively, the intermediate of formula (XXXV) may be treated with carbonyldπmidazole in a solvent such as dichloromethane, followed by freatment with the amme (XXXVII) to afford (XXXVIII) Alternatively, the intermediate of formula (XXXV) may be treated with HOBt, the amme (XXXVII), an organic base such as NMM, and a carbodπmide reagent such as EDC in a suitable solvent such as DMF, at a temperature of 0 °C to 80 °C to provide (XXXVIII) The compound of formula (XXXVIII) may be converted to (VI) by deprotection, conditions being particular to the nature of RPG2. For example, where RPG is tert-butoxycarbonyl, conversion of (XXXVIII) to (VI) may be accomplished by freatment of (XXXVIII) with TFA in the presence or absence of a suitable solvent such as dichloromethane, at a temperature of 0 °C to 50 °C.
Reaction scheme 7 depicts an alternate preparation of an intermediate of general formula (XXXV).
Fraction Scherre 7
Figure imgf000101_0001
R3 and R» are as defined for general formula (II).
RPG2 is as defined for reaction scheme 6.
R40 is lower alkyl or hydrogen. R41 is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, heterocyclylene, arylene, or heteroarylene, where alkylene, alkenylene, alkynylene, cycloalkylene, and cycloalkenylene substituents may contain one or more O, S, SO, or S02 substituents.
R 2 is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, heterocyclylene, arylene, O, NH, N-alkyl, or heteroarylene, where alkylene, alkenylene, alkynylene, cycloalkylene, and cycloalkenylene substituents may contain one or more O, S, SO, or S02 substituents.
R 3 is alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocyclyl, aryl, heteroaryl, or hydrogen, where alkyl, alkenyl, alkynyl, cycloalkyl, and cycloalkenyl substituents may contain one or more O, S, SO, or S02 substituents. R»4 is alkylene or heteroarylene. R45 is alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocyclyl, aryl, heteroaryl, or hydrogen, where alkyl, alkenyl, alkynyl, cycloalkyl, and cycloalkenyl substituents may contain one or more O, S, SO, or S02 substituents.
The hydroxy compound (XXXIX) may be treated with the reagent R43-R42-COCI in a solvent such as dichloromethane in the presence of tertiary base such as TEA to afford (XL). Alternately, (XXXIX) may be freated with R^-R^-COOH (where R,2 is not O, NH, or N- alkyl) and a dehydrating agent such as EDC and a catalyst such as DMAP in a solvent such as DMF or dichloromethane to afford (XL). The compound (XL) where R47 IS NH may be prepared by freating (XXXIX) with R43-NCO in a solvent such as dichloromethane. The ether (XLI) may be prepared by treating (XXXLX) with R^R-nBr or R45R44I in the presence of a base such as potassium carbonate or sodium hydride in a solvent such as DMF. Removal of the alkyl group R40 by saponification with aqueous base (or, if appropriate and where Rto is tert-butyl, by treatment with tπfluoroacetic acid) provides the acid (XXXV).
Reaction scheme 8 depicts an alternate preparation of an intermediate of general formula (XXXV).
Reaction Scheme 8
Figure imgf000103_0001
R3 and R4 are as defined for general formula (II). RPG2 is as defined for reaction scheme 6 R40, R4ι, ^4 and R45 are as defined for reaction scheme 7. n is 1 to 2.
The thioether (XLIII) may be prepared by treating (XLII) with tsR^Br or ^R^I and a base such as potassium carbonate or sodium hydride m a solvent such as DMF. The sulfur atom may be oxidized with a reagent such as m-chloroperoxybenzoic acid. Use of one molar equivalent of oxidant may be employed to provide (XLIV) where n is i . Use of two molar equivalents of oxidant may be employed to provide (XLIV) where n is 2. Removal of the alkyl group R,0 m either (XLIII) or (XLIV) by saponification with aqueous base (or, if appropriate and where R40 is tert-butyl, by treatment with tπfluoroacetic acid) provides the
Figure imgf000103_0002
Reaction scheme 9 depicts an alternate preparation of an intermediate of general formula (XXXV).
Reaction Scheme 9
Figure imgf000104_0001
R3 and R4 are as defined for general formula (II). RPG2 is as defined for reaction scheme 6.
Rto and R41 are as defined for reaction scheme 7.
Rtβ and R47 are, independently, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocyclyl, aryl, heteroaryl, carboxamidine, or hydrogen, where alkyl, alkenyl, alkynyl, cycloalkyl, and cycloalkenyl substituents may contain one or more O, S, SO, or S0 substituents. R46 and R47 may be taken together to constitute a three- to ten-membered πng.
The thiol (XLII) may be oxidized to the disulfide (XLV) by freatment with a mild base such as TEA and oxygen or air. Either the thiol (XLII) or the disulfide (XLV) may be converted to the sulfonyl chloride (XLVI) by treatment with chlorine gas in tetrachloromethane. Treatment of the sulfonyl chloride (XLVI) with an amme R46R47NH in the presence of a tertiary amme base such as TEA or NMM affords (XL VII). Removal of the alkyl group R40 in (XL VII) by sapomfication with aqueous base (or, if appropπate and where
R4o is tert-butyl, by treatment with frifluoroacetic acid) provides the acid (XXXV)
Reaction scheme 10 depicts an alternate preparation of an intermediate of general formula (XXXV).
Reaction Scheme 10
Figure imgf000105_0001
R3 and R are as defined for general formula (II).
RPG2 is as defined for reaction scheme 6.
R40 and R41 are as defined for reaction scheme 7. t6 and R47 are as defined for reaction scheme 9.
R^ and R47 may be taken together to constitute a three- to ten-membered πng.
The acid (XLVIII) may be converted to the amide (XLIX) by freatment of (XLVIII) and the a ine l^ ^NH with a dehydrating agent such as EDC or BOP in the presence of
HOBt. Removal of the alkyl group R40 in (XLIX) by sapomfication with aqueous base (or. if appropriate and where R 0 is tert-butyl, by treatment with frifluoroacetic acid) provides the acid (XXXV).
Reaction Scheme 11 depicts an alternate preparation of an intermediate of general formula
(XXXV). Reaction Scheme 11
Figure imgf000106_0001
R3 and R4 are as defined for general formula (II).
RPG2 is defined as for reaction scheme 6.
R48 is hydrogen, lower alkyl, or aryl.
R,9, R50, and R5] are, independently, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocyclyl, aryl, or heteroaryl. R50 and R5] may be taken together to constitute a five- to ten-membered πng.
R49 and R51 may be taken together to constitute a five- to ten-membered ring. R48 and R50 may be taken together to constitute a five- to ten-membered ring R52 is lower alkyl or benzyl. R53 is O or NH. L4 is Br or I.
The acid (L) is treated with frifluoroacetic anhydride and TFA to afford the tπfluoroacetamide, which is then treated with the bromide (LI) and a base such as potassium carbonate or DBU in a solvent such as DMF to provide the ester (LII). (LU) is treated with LDA and aluminum friisopropoxide in the presence of quinidme or quinine in a solvent such as THF at a temperature of from -78 °C to 25 °C to afford (LIII) with a high degree of asymmetric induction. (LIII) is subjected to hydrolysis with aqueous base and the resulting amme is protected with RPG2-C1 (where RPG2 is benzyloxycarbonyl) or (RPG2)20 (where RPG2 is tert-butoxycarbonyl) and aqueous base. (LIV) is then esteπfied with Rs2-L4 (where R52 is methyl or ethyl) and potassium carbonate in DMF or by treatment with dimethylformamide di-tert-butyl acetal (where R52 is tert-butyl). (LV) may be freated with ozone in dichloromethane or dichloromefhane/MeOH, followed by reduction with, for example, dimethyl sulfide to afford the carbonyl compound (LVI). (LVI) may be reduced with sodium borohydπde to afford the alcohol (LVIII) (where R53 is O), which may be freated in with methanesulfonyl chloride in pyridine to afford the mesylate. The methanesulfonate may be then freated with sodium azide in a solvent such as DMF at a temperature of from 25 °C to 120 °C to afford the azide, which may be reduced with, for example, palladium on barium sulfate and hydrogen gas to provide (LVIII) where R53 is NH. (LV) may be hydroborated with a borane reagent such as diborane followed by treatment with aqueous alkaline hydrogen peroxide to afford (LVII) where R53 is O (LVII) may be manipulated at
R53 m the manner described for (LVIII), and R53 may be acylated alkylated, or sulfonylated as desired. The R52 group may be removed by treatment with sodium hydroxide m aqueous THF
(where R52 is ethyl or methyl), or by freatment with TFA or anhydrous HC1 (where R52 is tert- butyl). Selection of R52 as tert-butyl and RPG2 as benzyloxycarbonyl is optimal for the preparation of (XXXV) according to reaction scheme 1 1.
Reaction scheme 12 depicts an alternate preparation of an intermediate of general formula (XXXV).
Reaction Scherre 12
Figure imgf000108_0001
R3 and R4 are as defined for general formula (II). RPG2 is as defined for reaction scheme 6.
R40 and R41 are as defined for reaction scheme 7.
R 6 and R47 are as defined for reaction scheme 9.
R^ and R47 may be taken together to constitute a three- to ten-membered ring.
R54 and R55 may be, independently, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocyclyl, aryl, heteroaryl, or hydrogen, where alkyl, alkenyl, alkynyl, cycloalkyl, and cycloalkenyl substituents may contain one or more O, S, SO, or S02 substituents. The amine compound (LLX) is treated with (LX) in the presence of a tertiary base such as TEA or NMM to afford (LXI). Treatment of (LXI) with silver nitrate and an amme HNR 6R47 provides (LXII). Removal of the alkyl group R 0 by sapomfication with aqueous base (or, if appropriate and where R40 is tert-butyl, by freatment with frifluoroacetic acid) provides (XXXV).
Reaction scheme 13 describes an alternate preparation of an intermediate of formula
(XXXV)
Reaction Scheme 13
Figure imgf000109_0001
LXI 11 LXIV
Figure imgf000109_0002
R3 and R4 are as defined for formula (II). RPG2 is as defined for reaction scheme 6. R40 is as defined for reaction scheme 7. R50 and R5] are as defined for reaction scheme 12.
The intermediate of formula (LXIII) may be treated with osmium tetraoxide or similar derivative m the presence of an oxidizing agent such as an iron (III) salt in aqueous tert-butanol to afford (LXIV). Asymmetric induction may be achieved in the reaction via use of a chiral ligand, such as hydroqumine 1 ,4-phthalazιnedιyl diether. (LXIV) may be treated with thionyl chloride and a tertiary amme base and the resulting sulfite may be oxidized with Ru04 or RuCl3 and NaI04 m aqueous acetonitπle/CCL, to afford the sulfate (LXV). Treatment of the sulfate with sodium azide m aqueous acetone affords (LXVI). Reduction of the azide with palladium on carbon and hydrogen gives the amine (LXVII). The amme may be alkylated with R3 if desired, protected with RPG2 (as described in previous schemes), and the R40 ester group removed to provide (XXXV). Selection of R40 as benzyl and RPG2 as tert-butoxycarbonyl are desirable for the sequence of reactions m reaction scheme 13.
Reaction scheme 14 describes an alternate preparation of an intermediate of formula (XXXV).
Reaction Scheme 14
Figure imgf000110_0001
R3 and R are as defined for general formula (II). RPG2 is as defined for reaction scheme 6.
R40, - and R 2 are as defined for reaction scheme 7.
R54 is as defined for reaction scheme 12.
R56 is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, heterocyclylene, arylene, O, NH, N-alkyl, or heteroarylene, where alkylene, alkenylene, alkynylene, cycloalkylene, and cycloalkenylene substituents may contain one or more O, S, SO, or S02 substituents.
R57 is alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocyclyl, aryl, heteroaryl, or hydrogen, where alkyl, alkenyl, alkynyl, cycloalkyl, and cycloalkenyl substituents may contain one or more O, S, SO, or S02 substituents.
The amme compound (LIX) may be freated with the reagent R57-R56-R 2-COCl m a solvent such as dichloromethane in the presence of tertiary base such as TEA to afford (LXVIII). Alternately, (LIX) may be freated with R57-R56- K-COOH (where R,2 is not O, NH, or N-alkyl) and a dehydrating agent such as EDC in a solvent such as DMF or dichloromethane to afford (LXVIII). The compound (LXVIII) where R42 is NH may be prepared by treating (LIX) with R57-R56-NCO m a solvent such as dichloromethane. Removal of the alkyl group R 0 by sapomfication with aqueous base (or, if appropriate and where R40 is tert-butyl, by treatment with frifluoroacetic acid) provides the acid (XXXV).
PHARMACEUTICAL FORMULATION AND DOSES
The compounds of the present invention can be administered m such oral (including buccal and sublmgual) dosage forms as tablets, capsules (each including timed release and sustained release formulations), pills, powders, granules, elixirs, tinctures, suspensions, syrups and emulsions. Likewise, they may also be administered m nasal, ophthalmic, otic, rectal, topical, intravenous (both bolus and infusion), mfrapeπtoneal, mtraarticular, subcutaneous or intramuscular inhalation or insufflation form, all using forms well known to those of ordinary skill m the pharmaceutical arts.
The dosage regimen utilizing the compounds of the present invention is selected accordance with a variety of factors including type, species, age, weight, sex and medical condition of the patient; the severity of the condition to be treated; the route of administration: the renal and hepatic function of the patient; and the particular compound or salt thereof employed. An ordinarily skilled physician or veterinarian can readily determine and prescribe the effective amount of the drug required to prevent, counter or arrest the progress of the condition.
Oral dosages of the present invention, when used for the indicated effects, will range between about 0.1 to 2000 mg kg of body weight per day, and particularly 1 to 100 mg/kg of body weight per day. Oral dosage units will generally be administered in the range of from 1 to about 250 mg and more preferably from about 25 to 250 mg The daily dosage for a 70 kg mammal will generally be in the range of about 10 mg to 5 grams of a compound of formula (I) or (II).
While the dosage to be administered is based on the usual conditions such as the physical condition of the patient, age, body weight, past medical history, route of administrations, severity of the conditions and the like, it is generally preferred for oral administration to administer to a human. In some cases, a lower dose is sufficient and, in some cases, a higher dose or more doses may be necessary. Topical application similarly may be once or more than once per day depending upon the usual medical considerations. Advantageously, compounds of the present invention may be administered in a single daily dose, or the total daily dosage may be administered in divided doses of two, three or four times daily. Furthermore, preferred compounds for the present invention can be administered in infranasal form via topical use of suitable mtranasal vehicles, or via transdermal routes, using those forms of transdermal skin patches well known to those of ordinary skill in that art. To be administered in the form of a transdermal delivery system, the dosage administration will, of course, be continuous rather than intermittent throughout the dosage regimen.
In the methods of the present invention, the compounds herein described m detail can form the active ingredient, and are typically administered in admixture with suitable pharmaceutical diluents, excipients or carriers (collectively referred to herein as "earner" materials) suitably selected with respect to the intended form of administration, that is, oral tablets, capsules, elixirs, syrups and the like, and consistent with conventional pharmaceutical practices.
For instance, for oral administration in the form of a tablet or capsule, the active drug component can be combined with an oral, non-toxic pharmaceutically acceptable inert earner such as ethanol, glycerol, water and the like Powders are prepared by comminuting the compound to a suitable fine size and mixing with a similarly comminuted pharmaceutical carrier such as an edible carbohydrate, as, for example, starch or manmtol. Flavoring, preservative, dispersing and coloring agent can also be present.
Capsules are made by preparing a powder mixture as described above, and filling formed gelatin sheaths. Ghdants and lubricants such as colloidal silica, talc, magnesium stearate, calcium stearate or solid polyethylene glycol can be added to the powder mixture before the filling operation. A disintegrating or solubihzing agent such as agar-agar, calcium carbonate or sodium carbonate can also be added to improve the availability of the medicament when the capsule is ingested.
Moreover, when desired or necessary, suitable binders, lubricants, disintegrating agents and coloring agents can also be incorporated into the mixture. Suitable binders include starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, fragacanth or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes and the like. Lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like. Disintegrators include, without limitation, starch, methyl cellulose, agar, bentonite, xanthan gum and the like. Tablets are formulated, for example, by preparing a powder mixture, granulating or slugging, adding a lubricant and dis tegrant and pressing into tablets. A powder mixture is prepared by mixing the compound, suitably comminuted, with a diluent or base as described above, and optionally, with a binder such as carboxymethylcellulose, an ahginate, gelatin, or polyvmyl pyrrolidone, a solution retardant such as paraffin, a resorption accelerator such as a quaternary salt and/or an absorption agent such as bentonite, kaolm or dicalcium phosphate The powder mixture can be granulated by wetting with a binder such as syrup, starch paste, acadia mucilage or solutions of cellulosic or polymeric materials and forcing through a screen. As an alternative to granulating, the powder mixture can be run through the tablet machine and the result is imperfectly formed slugs broken into granules. The granules can be lubricated to prevent sticking to the tablet forming dies by means of the addition of stearic acid, a stearate salt, talc or mineral oil. The lubricated mixture is then compressed into tablets. The compounds of the present invention can also be combined with free flowing inert carrier and compressed into tablets directly without going through the granulating or slugging steps A clear or opaque protective coating consisting of a sealing coat of shellac, a coating of sugar or polymeric material and a polish coating of wax can be provided. Dyestuffs can be added to these coatings to distinguish different unit dosages.
Oral fluids such as solution, syrups and elixirs can be prepared in dosage unit form so that a given quantity contains a predetermined amount of the compound Syrups can be prepared by dissolving the compound in a suitably flavored aqueous solution, while elixirs are prepared through the use of a non-toxic alcoholic vehicle. Suspensions can be formulated by dispersing the compound in a non-toxic vehicle. Solubihzers and emulsifiers such as ethoxylated isostearyl alcohols and polyoxy ethylene sorbitol ethers, preservatives, flavor additive such as peppermint oil or saccharin, and the like can also be added.
Where appropπate, dosage unit formulations for oral administration can be microencapsulated. The formulation can also be prepared to prolong or sustain the release as for example by coating or embedding particulate material polymers, wax or the like. The compounds of the present invention can also be administered in the form of l posome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles. Liposomes can be formed from a variety of phosphohpids, such as cholesterol, stearylamine or phosphatidylcholmes.
Compounds of the present invention may also be delivered by the use of monoclonal antibodies as individual carriers to which the compound molecules are coupled. The compounds of the present invention may also be coupled with soluble polymers as targetable drug carriers. Such polymers can include polyvinylpyrrohdone, pyran copolymer, polyhydroxypropylmethacrylamide-phenol, polyhydroxyethylaspartamidephenol, or polyethyleneoxidepolylysme substituted with palmitoyl residues. Furthermore, the compounds of the present invention may be coupled to a class of biodegradable polymers useful m achieving controlled release of a drug, for example, polylactic acid, polepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates and cross-linked or amphipafhic block copolymers of hydrogels.
The present invention includes pharmaceutical compositions containing 0.1 to 99.5%, more particularly, 0.5 to 90% of a compound of the formula (II) in combination with a pharmaceutically acceptable carrier.
Parenteral administration can be effected by utilizing liquid dosage unit forms such as sterile solutions and suspensions intended for subcutaneous, intramuscular, lntrathecal, mfraarteπal or intravenous injection. These are prepared by suspending or dissolving a measured amount of the compound in a non-toxic liquid vehicle suitable for injection such as aqueous oleaginous medium and sterilizing the suspension or solution. Alternatively, a measured amount of the compound is placed m a vial and the vial and its contents are sterilized and sealed. An accompanying vial or vehicle can be provided for mixing prior to administration. Non-toxic salts and salt solutions can be added to render the injection lsotonic. Stabilizers, preservations and emulsifiers can also be added.
Rectal administration can be effected utilizing suppositories which the compound is admixed with low-meltmg water-soluble or insoluble solids such as polyethylene glycol, cocoa butter, higher ester as for example flavored aqueous solution, while elixirs are prepared through myπstyl palmitate or mixtures thereof.
Topical formulations of the present invention may be presented as, for instance, ointments, creams or lotions, eye ointments and eye or ear drops, impregnated dressings and aerosols, and may contain appropriate conventional additives such as preservatives, solvents to assist drug penetration and emollients in ointments and creams. The formulations may also contain compatible conventional carriers, such as cream or ointment bases and ethanol or oleyl alcohol for lotions. Such carriers may be present as from about 1% up to about 98% of the formulation More usually they will form up to about 80% of the formulation.
For administration by inhalation the compounds according to the invention are conveniently delivered m the form of an aerosol spray presentation from pressurized packs or a nebulizer, with the use of a suitable propellant, e.g. dichlorodifluoromethane, tπchlorofluoromefhane, dichlorotetrafluoroethane, tetrafluoroethane, heptafluoropropane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol the dosage unit may be determined by providing a valve to deliver a metered amount. Capsules and cartridges of e.g. gelatin for use in an inhaler or insufflator may be formulated containing a powder mix of a compound of the invention and a suitable powder base such as lactose or starch. The prefeπed pharmaceutical compositions are those in a form suitable for oral administration, such as tablets and liquids and the like and topical formulations.
EXAMPLES
The following examples are illustrative embodiments of the invention, not limiting the scope of the invention in any way. Reagents are commercially available or are prepared according to procedures m the literature.
Example 1; (2R,3S)-3-(Formyl-hydroxyammo)-2-(3-phenyl-l-propyl)butanoιc Acid (lS)-2,2- Dimethyl- 1 -(3-pyπdylcarbamoyl)- 1 -propyl] amide
Figure imgf000117_0001
Example la; Methyl (2R,3R)-2-[(2£)-3-Phenyl-2-propen-l-yl]-3-hydroxybutanoate
To a solution of dnsopropylamme (47.1 g, 466.1 mmol) in THF (500 mL) cooled to -50 °C is added n-butylhfhium (466.1 mmol, 2.5M in hexanes) and the resulting solution is stirred at -50 °C for 0.5 h. The reaction mixture is cooled to -78 °C followed by slow addition of methyl (3R)-3-hydroxybutanoate (25 g, 211.9 mmol). After 0.5 h a solution of cmnamyl bromide (45.9 g, 233.0 mmol) in HMPA (10 mL) is added and the reaction mixture is allowed to warm to 0 °C and stirred for 16 h. The reaction mixture is quenched by addition 30 mL of saturated aqueous ammonium chloride solution, is poured into 400 mL of 1 N hydrochloric acid, and is extracted with two 500 - mL portions of EtOAc. The combined organic layers are dried over magnesium sulfate, concentrated in vacuo, and purified by silica gel chromatography (elution with 25% EtOAc - hexanes) to afford methyl (2R,3R)-2-[(2£)-3-phenyl-2-propen-l-yl]-3-hydroxybutanoate as a yellow oil (42 g, 85% yield). Η NMR (400 MHz, CDCI3) δ 7.32-7.18 (m, 5H), 6.44 (d, 1H), 6 13 (m, 1H), 3.98 (m, 1H),
3.69 (s, 3H), 2.56 (m, 4H), 1.25 (t, 3H) ppm. ESI-MS m/z 257 (M+Na)+.
Example lb; Methyl (2R,3R)-2-(3-Phenyl-l-propyl)-3-hydroxybutanoate
A solution of methyl (2R,3R)-2-[(2£')-3-phenyl-2-propen-l-yl]-3- hydroxybutanoate (42.0 g, 179.5 mmol) in 400 mL of methanol is treated with 400 mg of 10% palladium on carbon. The resulting suspension is repeatedly evacuated and purged with a hydrogen balloon, then stirred under 1 atmosphere pressure of hydrogen gas for 16 h. The catalyst is filtered and the filtrate is concenfrated in vacuo to provide methyl (2R,3R)-2-(3- phenyl-l-propyl)-3-hydroxybutanoate as an oil (42.2 g, 100% yield). Η NMR (400 MHz, CDCI3) δ 7.25 (m, 2H), 7.17 ( , 3H), 3.88 (m, 1H), 3.69 (s, 3H), 2.61
(m, 2H), 2.42 (m, 2H), 1.72 (m, 1H), 1.62 (m, 3H), 1 19 (d, 3H) ppm. ESI-MS m/z 259 (M+Na)+.
Example lc; (2R,3R)-2-(3-Phenyl-l-propyl)-3-hydroxybutanoιc Acid
To a solution of methyl (2R,3R)-2-(3-phenyl-l-propyl)-3-hydroxybutanoate (42.2 g, 179.5 mmol) in THF - methanol (3.1, 535 mL) is added 2 N aqueous sodium hydroxide solution (135 mL, 269.3 mmol). The solution is stirred at 23 °C for 20 h, then concentrated and extracted with hexanes (100 mL). The aqueous layer is acidified to pH 3 with saturated aqueous sodium bisulfate and is extracted with two 500 - mL portions of EtOAc. The combined organic layers are dried over magnesium sulfate and concentrated under reduced pressure to provide (2R,3R)-2-(3-phenyl-l-propyl)-3-hydroxybutanoιc acid as an oil (33.0 g, 83% yield).
Η NMR (400 MHz, CDCI3) δ 7.28 (m, 2H), 7.16 (m, 3H), 3.93(m, IH), 2.63 (m, 2H), 2.43
(m, IH), 1.69 (m, 4H), 1.26 (d, 3H) ppm. ESI-MS m/z 221 (M-l)\
Example Id; (2R,3R)-2-(3-Phenyl-l-propyl)-3-hydroxybutanoιc Acid 2- Tefrahydropyranyloxyamide
To a solution of (2R,3R)-2-(3-phenyl-l-propyl)-3-hydroxybutanoιc acid (33.0 g, 148.7 mmol) in dichloromethane (300 mL) is added 2-tetrahydropyranyloxyamιne (18.3 g, 156.1 mmol) and EDC (31.2 g, 163.5 mmol). The resulting solution is stirred at 23 °C for 16 h, then diluted with dichloromethane (500 mL) and washed sequentially with 1 N hydrochloric acid, saturated aqueous sodium bicarbonate solution, and saturated aqueous sodium chloride solution. The reaction mixture is dried over magnesium sulfate and concentrated under reduced pressure to provide (2R,3R)-2-(3-phenyl-l-propyl)-3- hydroxybutanoic acid 2-tefrahydropyranyloxyamιde as a foam (47.8 g, 100% yield). Η NMR (400 MHz, CDCI3) δ 8.66 (bs, IH), 7.25 (m, 2H), 7.17 (m, 3H), 4.94 (m, IH), 3.89
(m, 2H), 3.61 (m, IH), 2.62 (t, 2H), 1.93 (m, IH), 1.78 (m, 4H), 1.66 (m, 6H), 1.23 (d, 3H) ppm. ESI-MS m/z 320 (M-l)\
Example 1 e; (3R,4S)- 1 -(2-Tetrahydropyranyloxy)-3 -(3 -phenyl- 1 -propyl)-4-methylazetιdm-2- one To a solution of (2R,3R)-2-(3-phenyl-l-propyl)-3-hydroxybutanoιc acid 2- tetrahydropyranyloxyamide (47.8 g, 148.7 mmol) in 150 mL of dichloromethane at 0 °C is added pyridine (64 mL) and methanesulfonyl chloride (20.4 g, 178 4 mmol). The resulting solution is allowed to warm to 23 °C and is stirred at 23 °C for 14 h, concentrated in vacuo. and diluted with dichloromethane (500 mL). The organic layer is washed with 1 N hydrochloric acid, saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, and concenfrated to provide the methanesulfonate intermediate.
A suspension of potassium carbonate (61.5 g) in acetone (500 mL) is heated to reflux for 1 h. A solution of the above methanesulfonate m acetone (1 L) is added and the resulting suspension is heated at reflux for 28 h. The mixture is allowed to cool to 25 °C and is filtered, concentrated under reduced pressure, and purified by silica gel chromatograph) (elution with 3:1 hexanes - EtOAc) to provide (3R,4S)-l-(2-tetrahydropyranyloxy)-3-(3- phenyl-l-propyl)-4-methylazetιdm-2-one as an oil (34.0 g, 75% yield). Η NMR (400 MHz, CDCI3) δ 7.25 (m, 2H), 7.17 (m, 3H), 5.13 and 4.99 (two m, IH), 4.15-
3.98 (m, 2H), 3.64 (m, IH), 2.93 (m, IH), 2.67 (m, 2H), 1.89-1.51 (m, 10H), 1.28 and 1.26 (two d, 3H) ppm. ESI-MS m/z 326 (M+Na)+.
Example If; (2R,3S)-2-(3-Phenyl-l-propyl)-3-(2-tetrahydropyranyloxyammo)butanoιc Acid
To a solution of (3R,4S)-l-(2-tetrahydropyranyloxy)-3-(3-phenyl-l-propyl)-4- mefhylazetιdιn-2-one (20.5 g, 67.6 mmol) in dioxane (220 mL) is added 1 N aqueous sodium hydroxide (102 mL). The solution is stirred at 23 °C for 20 h, then extracted with hexanes (200 mL). The aqueous layer is acidified to pFI 3 with saturated aqueous sodium bisulfate solution, and is extracted with two 300 - mL portions of EtOAc. The combined organics are washed with saturated aqueous sodium chloride, dried over anhydrous magnesium sulfate. and concentrated under reduced pressure to provide (2R,3S)-2-(3-phenyl-l-propyl)-3-(2- tetrahydropyranyloxyammo)butanoιc acid as an oil (21.5 g, 99% yield)
Η NMR (400 MHz, CDCI3) δ 7.28 (m, 2H), 7.18 (m, 3H), 4 84 and 4.70 (two m, IH), 3.96
and 3.89 (two m, IH), 3.56 (m, IH), 3.34 and 3.24 (two m, IH), 2.97 and 2.81 (two m, IH), 2.65 (m, 2H), 1.96-1.45 (m, 10H), 1.31 (m, IH), 1.06 and 0.99 (two d, 3H) ppm. ESI-MS m/z 344 (M+H)+.
Example lg; (2R,3S)-2-(3-Phenyl-l-propyl)-3-(formyl-2- tetrahydropyranyloxyamιno)butanoιc Acid
To a solution of (2R,3S)-2-(3-phenyl-l-propyl)-3-(2- tefrahydropyranyloxyamιno)butanoιc acid (21.4 g, 66.7 mmol) in pyπdine (100 mL) at 0 °C is added formic acetic anhydride ( 30 mL). The resulting solution is allowed to warm to 25 °C, stirred for 6 h, and then concentrated to dryness under reduced pressure. The resulting gum is dissolved m EtOAc (300 mL) and washed sequentially with 1 N hydrochloric acid (200 mL) and saturated aqueous sodium chloride solution. The organic layer is dried over anhydrous magnesium sulfate and concentrated under reduced pressure providing (2R,3S)-2-(3-phenyl-l- propyl)-3-(formyl-2-tetrahydropyranyloxyamιno)butanoιc acid as an oil (18.9 g, 81% yield). ESI-MS m z 372 (M+Na)+ 348 (M-l)".
Example lh; (2S)-2-tert-Butoxycarbonylamιno-3,3-dιmethylbutanoιc Acid 3-Pyπdylamιde
To a solution of (2S)-2-tert-butoxycarbonylammo-3,3-dιmethylbutanoιc acid (3 g, 12.99 mmol) in DMF (25 mL) is added EDC (2.73 g, 14.29 mmol) and HOBT (1.93 g, 14.29 mmol). The resulting solution is stirred at 25 °C for 0.5 h and 3-amιnopyπdιne (1.83 g, 19.48 mmol) is added and the reaction is heated to 50 °C for 18 h. The mixture is concentrated, diluted with EtOAc (50 mL), and washed with saturated aqueous sodium bicarbonate. The organic layer is dried over anhydrous magnesium sulfate and concenfrated in vacuo to provide (2S)-2-tert-butoxycarbonylamιno-3,3-dιmethylbutanoιc acid 3- pyπdylamide as a foam (1.6 g, 40% yield).
Η NMR (400 MHz, CDCI3) δ 8.56 (s, IH), 8.24 (bs, IH), 7.99 (bs, IH), 7.18 (bs, IH), 5.16
(m, IH), 4.06 (m, IH), 1.43 (s, 9H), 1.02 (s, 9H) ppm. ESI-MS m/z 308 (M+H)+.
Example ; (2S)-2-Amιno-3,3-dιmethylbutanoιc Acid 3-pyπdylamιde Dihydrochloπde
To a solution of (2S -2-tert-butoxycarbonylammo-3,3-dιmethylbutanoιc acid 3- pyπdylamide (1.6 g, 5.21 mmol) in dichloromethane (10 mL) cooled at 0 °C is added 4 M HCl m dioxane (10 mL). The resulting solution is allowed to warm to 25 °C and is stirred for 3 h. The reaction mixture is concentrated and the solid is filtered to provide (2S)-2-amιno- 3,3-dιmethylbutanoιc acid 3-pyπdylamιde dihydrochloπde (1.40 g, 94% yield) as a white solid.
Η NMR (400 MHz, CD3OD) δ 9.44 (s, IH), 8.69 (d, IH), 8.64 (d, IH), 8.09 (dd, IH), 3.96 (s, IH), 1.17 (s, 9H) ppm. ESI-MS m/z 208 (M+H)+.
Example lj; (2R,3S)-3-(Formyl-2-tetrahydropyranyloxyamιno)-2-(3-phenyl-l- propyl)butanoιc Acid [(lS)-2,2-Dιmethyl- 1 -(3-pyπdylcarbamoyl)- 1 -propyl]amιde
To a solution of (2R,3S)-2-(3-phenyl-l-propyl)-3-(formyl-2- tefrahydropyranyloxyamιno)butanoιc acid (120 mg, 0.344 mmol) in DMF (1 mL) is added BOP reagent (167 mg, 0.378 mmol), HOBt ( 51 mg, 0.378 mmol), and NMM (174 mg, 1.72 mmol). After 30 min, (2S)-2-amιno-3,3-dιmethylbutanoιc acid 3-pyπdylamιde dihydrochloπde (118 mg, 0.410) is added and the resulting solution is stirred at 25 °C for 18 h. The reaction mixture is concentrated, diluted with EtOAc (20 mL) and washed sequentially with 1 M aqueous sodium carbonate and saturated aqueous sodium chloride. The organic layer is dried over magnesium sulfate, concentrated in vacuo, and purified by silica gel chromatography (elution with 2.1 EtOAc - hexane) to provide (2R,3S)-3-(formyl-2- tetrahydropyranyloxyammo)-2-(3-phenyl-l-propyl)butanoιc acid [(lS)-2,2-dιmethyl-l-(3- pyπdylcarbamoyl)-l-propyl]amιde as a solid (115 mg, 62% yield) ESI-MS m/z 539 (M+H)+.
Example 1 , (2R,3S)-3-(Formyl-hydroxyamιno)-2-(3-phenyl-l-propyl)butanoιc Acid [(lS)-2,2- Dimethyl- l-(3-pyπdylcarbamoyl)-l-propyl]amιde
A solution of (2R,3S)-3-(formyl-2-tetrahydropyranyloxyamιno)-2-(3-phenyl-l- propyl)butanoιc acid [(lS)-2,2-dιmethyl-l-(3-pyπdylcarbarnoyl)-l-propyl]amιde (115 mg, 0.214 mmol) in acetic acid - water (4: 1, 1 mL) is heated to 50 °C for 18 h. The reaction mixture is concentrated, then dissolved in toluene and concentrated in vacuo. The procedure is repeated once again to afford the crude product which is triturated from hot dichloromethane - ether to provide (2R,3S)-3-(formyl-hydroxyamιno)-2-(3-phenyl-l- propyl)butanoιc acid [(lS)-2,2-dιmethyl-l-(3-pyndylcarbamoyl)-l-propyl]amιde as an off- white solid (70 mg, 72% yield). Η NMR (400 MHz, CD3OD) δ 8.76 (s, IH), 8.25 and 7.99 (two s, IH), 8.22 (d, IH), 8,02 (t, IH), 7.36 (dd, IH), 7.00 (m, 5H), 4.48 and 3.82 (two dq, IH), 4.44 (s, IH), 2.94 and 2.82 (two m, IH), 2.58 (m, IH), 2.42 (m, IH), 1.50 (m, 4H), 1.24 and 1.18 (two d, 3H), 1.02 (s, 9H) ppm. ESI-MS m z 455 (M+H)+. Anal. Calcd. for C25H34N4θ4 0.5 CH3C02H: C, 64 44; H, 7.49, N, 11.56. Found. C, 64.52; H, 7.49; N, 1 1.48. Example 2; (2R,3S)-3-(Formyl-hydroxyammo)-2-(4-phenylcyclohexylmethyl)butanoιc Acid
[(lS)-2,2-Dιmethyl-l-(l,3-thιazol-2-ylcarbamoyl)-l-propyl]amιde
Figure imgf000124_0001
Example 2a; 4-Phenyl- 1 -methylenecyclohexane
μ-Chloro-μ-methylene[bιs(cyclopentadιenyl)tιtanιum]dιmethylalummum (172 mL, 0.086 mol, 0.5 M in toluene) is added dropwise to a solution of 4-phenylcyclohexanone (15.0 g, 0.086 mol) in THF (100 mL) at 0 °C. The mixture is allowed to come to 25 °C. After 15 min., ether (100 mL) is added followed by dropwise addition of 0.1 M sodium hydroxide (120 mL). The mixture is stirred an 25 °C for 18 h, filtered, and the filtrate is dπed over sodium sulfate and concentrated in vacuo . The residue is purified by column chromatography on silica gel using dichloromethane - hexanes (1 :1) as eluent. The resulting yellow oil is rechromatographed using hexanes as eluent to give 8.9 g (61%) of 4-phenyl-l- mefhylenecyclohexane as an oil. 'H NMR (300 MHz, CDC13) δ 7.36-7.23 (m, 5H), 4.72 (s, 2H), 2.71 (m, IH), 2.47 (m, 2H), 2.22 (m, 2H), 2.03 (m, 2H), 1.54 (m, 2H) ppm. ESI-MS m/z 173 (M+H)+.
Example 2b; 4-Phenylcyclohexylmethanol.
A solution of 4-phenyl-l -methylenecyclohexane (8.9 g, 0.051 mol) in THF (10 mL) is added dropwise to a solution of borane (15 mL, 0.015 mol, 1 M m THF) at 0 °C under argon. The mixture is stirred at 25 °C for 2.5 h. A solution of aqueous sodium hydroxide (5 mL, 0.015 mol, 3 M) is added dropwise, the mixture is chilled to 0 °C and aqueous hydrogen peroxide (6 mL, 0.06 mol, 30%) is added dropwise. After stirring at 25 °C for 1 h the mixture is poured into water and extracted with ether. The organic phase is dried over sodium sulfate and concentrated in vacuo to give 9.9 g of a colorless oil. The oil is purified by column chromatography on silica gel using hexanes - EtOAc (2: 1) as eluent to give 7.0 g (72%) of 4- phenylcyclohexylmethanol as an oil.
Η NMR (300 MHz, CDC13) δ 7.36-7.20 (m, 5H), 3.75 (m, IH), 3.56 (m, IH), 2.59 (m, IH), 2.08-1.15 (m, 10H) ppm
Example 2c, 4-Phenylcyclohexylmethyl Iodide
Imidazole (4.4 g, 0.065 mol) is added in one portion to a solution of tπphenylphosphme (17.0 g, 0.065 mol) m dichloromethane (300 mL) at 25 °C . After all of the solids dissolved, iodine (16.5 g, 0.065 mol) is added in four portions. The mixture is stirred for 10 min, then a solution of 4-phenylcyclohexylmethanol (10.5 g, 0.055 mol) is added dropwise and the mixture is stirred at 25 °C for 18 h. The reaction mixture is poured into pentane and the resulting solids filtered. The filtrate is concentrated in vacuo and the residue is dissolved m dichloromethane. This solution is poured into hexanes, the solids were filtered, and the filtrate is concentrated in vacuo. The residue is passed through a pad of silica gel using hexanes as eluent to afford 14.6 g (89%) of 4-phenylcyclohexylmethyl iodide
Η NMR (400 MHz, CDC13) δ 7.32-7.17 (m, 5H), 3.35 (dd, IH), 3.17 (dd, IH), 2.60 and 2.47 (two m, IH), 2.05 (m, IH), 1.95-1.84 (m, 2H), 1.77-1.64 (m, 3H), 1.56-1.47 (m, 2H), 1.18 (m, IH) ppm.
Example 2d; Methyl (2R,3R)-3-Hydroxy-2-(4-phenylcyclohexylmethyl)butanoate A solution of methyl (3R)-3-hydroxybutyrate (5.6 g, 0.047 mol) in THF (20 mL) is added dropwise to a solution of LDA prepared by dropwise addition of n-butylhthium (49.5 mL, 0.099 mol, 2 M in cyclohexane) to dnsopropylamme (10 0 g, 0.099 mol) in THF (20 mL) at -78 °C. After stirring at -50 °C to -40 °C for 0.5 h, a solution of 4- phenylcyclohexylmethyl iodide (14 6 g, 0.049 mol) m a mixture of HMPA (10 mL) and THF (15 mL) is added dropwise. The mixture is kept at -20 °C for 48 h, then allowed to warm to 4 °C and kept at that temperature for 48 h. The reaction mixture is chilled to 0 °C and quenched by dropwise addition of saturated aqueous ammonium chloride solution. The aqueous phase is extracted with ether and the combined organic phase is washed with ice-cold 0.1 M HCl, then saturated sodium chloride solution and dried over sodium sulfate. Concentration of the organic phase gives 14.6 g of crude product Column chromatography on silica gel with hexanes - EtOAc (2: 1) as eluent affords 4.18 g (31%) of methyl (2R,3R)-3-hydroxy-2-(4- phenylcyclohexylmethyl)butanoate as a pale yellow oil which solidified on standing. Η NMR (300 MHz, CDC13) δ 7.35-7.22 (m, 5H), 3.93 (m, IH), 3.78 (s, 3H), 3.52 (m, 4H), 2.09-1.64 (m, 5H), 1.55-1.30 (m, 3H), 1.28 (d, 3H), 1.21-1.00 (m, 2H) ppm. Anal. Calcd. for C,8H2603: C, 74 45; H, 9.03. Found. C, 74.52; H, 8.96.
Example 2e; (2R,3R)-3-Hydroxy-2-(4-phenylcyclohexylmethyl)butanoιc Acid
A mixture of methyl (2R,3R)-3-hydroxy-2-(4-phenylcyclohexylmethyl)butanoate
(4.17 g, 0.014 mol) and lithium hydroxide monohydrate (0.59 g, 0.014 mol) is stirred at 25 °C m a mixture of THF (30 mL), methanol (15 mL) and water (15 mL) for 72 h. The reaction is not complete and an additional 0.88 g (0.021 mol) of lithium hydroxide monohydrate m water (10 mL) is added. After 6 h an additional 0.59 g (0.014 mol) of solid lithium hydroxide monohydrate and the mixture is stirred at 25 °C for 18 h. The mixture is concentrated in vacuo and the residue is taken up in water, chilled in an ice-bath and the pH is adjusted to 2 with sodium bisulfate. The mixture is extracted with dichloromethane. The combined extracts were washed with saturated sodium chloride, dried over sodium sulfate and concentrated in vacuo to give 3.43g (89%) of (2R,3R)-3-hydroxy-2-(4-phenylcyclohexylmethyl)butanoιc acid as a yellow solid.
Η NMR (300 MHz, DMSO-d6) δ 11.9 (br, IH), 7.31-7.17 (m, 5H), 4.80 (br, IH), 3.75 (m, IH), 2.53-2.30 (m, 2H), 1.98-0.90 ( , 1 IH), 1 08 (d, 3H) ppm.
Example 2f; (2R,3R)-3-Hydroxy-2-(4-phenylcyclohexylmethyl)butanoιc Acid 2- Tefrahydropyranyloxyamide
A mixture of (2R,3R)-3-hydroxy-2-(4-phenylcyclohexylmethyl)butanoιc acid (3.43 g, 0.012 mol), 2-tetrahydropyranyloxyamιne (1 64 g, 0.014 mol), BOP (5.75 g, 0.013 mol), HOBt (1.76 g, 0.13 mol) and NMM (2.63 g, 0.026 mol) m DMF (30 mL) is stirred at 25 °C for 18 h. The mixture is concenfrated in vacuo and the residue is taken up in EtOAc. The mixture is washed with ice-cold 0.1 M HCl, saturated aqueous sodium bicarbonate, and saturated aquesous sodium chloride. The organic phase is dried over sodium sulfate and concentrated in vacuo to give 6.02 g of a white solid. Chromatography on silica gel using EtOAc as eluent gives 4.05 g (90%) of (2R,3R)-3-hydroxy-2-(4- phenylcyclohexylmethyl)butanoιc acid 2-tetrahydropyranyloxyamιde as a white solid. Η NMR (400 MHz, DMSO-d6) δ 10.90 (m, IH), 7.25-7.10 ( m, 5H), 4.81 (m, IH), 4.53 (t, IH), 3.90 (m, IH), 3.61 (m, IH), 3.45 (m, IH), 2.39 (br t,lH), 2.09 (m, IH), 1.95 (m, IH), 1.73-0.85 (m, 16H), 1.01 (d, 3H) ppm.
Example 2g; (3R,4S)-l-(2-Tetrahydropyranyloxy)-3-(4-phenylcyclohexylmethyl)-4- methylazetιdιn-2-one
Methanesulfonyl chloride (1.4 g, 0.012 mol) is added dropwise to a solution of
(2R,3R)-3-hydroxy-2-(4-phenylcyclohexylmethyl)butanoιc acid 2-tetrahydropyranyloxyamιde (4.02 g, 0.01 1 mol) in pyridine (30 mL) at 0 °C. After 3 h at 25 °C the mixture is concenfrated in vacuo and the residue is taken up in a mixture of EtOAc and cold 1M HCl. The organic phase is washed with cold 1 M HCl and saturated sodium chloride solution. The organic phase is dried over sodium sulfate and concentrated in vacuo to give the crude methanesulfonate as a white foam Η NMR (400 MHz, DMSO-d6) δ 11.25 (d, IH), 7.25-7.12 (m, 5H), 4.86 (br s, IH). 4.63 (m,
IH), 4.10 (m, 2H), 3.98 (m,lH), 3.02 (d, 3H), 2.41 (m, 2H), 2.00 (m, IH), 1.74-0.80 (m,
15H), 1.33 (dd, 3H) ppm.
A solution of the above mesylate (5.0 g, 0.011 mol) m acetone (30 mL) is added dropwise to a mixture of potassium carbonate (4.7 g, 0.034 mol) in refluxing acetone (100 mL). After refluxing for 18 h, the mixture is filtered and the filtrate is concentrated in vacuo. The residue is purified by column chromatography on silica gel with hexanes -
EtOAc(l : l) as eluent to give 3.85 g (98%) of (3R,4S)-l-(2-tetrahydropyranyloxy)-3-(4- phenylcyclohexylmethyl)-4-methylazetιdm-2-one as an oil.
Η NMR (300 MHz, DMSO-d6) δ 7.32-7.18 (m, 5H), 5.05 (d, IH), 4.06 (m, 2H), 3.57 (br t, IH), 3.10 (m, IH), 2.52 (m, IH), 1.95-1.39 (m, 15H), 1.24 (dd, 3H), 1.09 (m, 2H) ppm.
ESI-MS m/z 358 (M+H)+
Anal. Calcd. for C22H3,N03: C, 73.91; H, 8.74; N, 3.92. Found: C, 73.66; H, 8.77; N, 3.89.
Example 2h; (2R,3S)-2-(4-phenylcyclohexylmethyl)-3-(2- tetrahydropyranyloxyammo)butanoιc acid
A solution of 3 N sodium hydroxide (18.7 mL) is added dropwise to a solution of (3R,4S)-1- (2-tefrahydropyranyloxy)-3-(4-phenylcyclohexylmethyl)-4-methylazetιdm-2-one (3.6 g, 0.010 mol) m dioxane (50 mL) at 25 °C and the mixture is stirred for 18 h. Water (10 mL) is added to the reaction mixture and an additional 5 mL of 3 N sodium hydroxide is added dropwise and the mixture is stirred an additional 8 h. The reaction mixture is mixed with water (200 mL), 1 M sodium hydrogen sulfate (60 mL) and EtOAc (100 mL). The aqueous phase is extracted with EtOAc. The combined organic phase is washed with water and saturated sodium chloride solution, dried (sodium sulfate) and concentrated in vacuo to give 3.44 g (92%) of (2R,3S)-2-(4-phenylcyclohexylmethyl)-3 -(2-tetrahydropyranyloxyammo)butanoιc acid as a white solid. The product is used without further purification: 'H NMR (300 MHz, DMSO-d6) δ 12.07 (br, IH), 7.28-7.17 (m, 5H), 6.52 (br, IH), 4.70 (br d, IH), 3.81 (m, IH), 3.45 (m, IH), 3.05 (m, IH), 2.55 (m, 2H), 1.93-1.12 (m, 17H), 1.03 (dd, 3H) ppm. ESI-MS m/z 374 (M-H) .
Example 2ι; (2R,3S)-3-(Formyl-2-tetrahydropyranyloxyammo)-2-(4- phenylcyclohexylmethyl)butanoιc acid
Formic acetic anhydride (2.03 g, 0.023 mol) is added via syringe to a mixture of (2R,3S)-2-(4-phenylcyclohexylmethyl)-3-(2-tefrahydropyranyloxyammo)butanoιc acid (3.44 g, 0.009 mol) in pyridme (35 mL) at 0 °C. The mixture is allowed to come to 25 °C. Additional formic acetic anhydride (0.97g, 0.011 mol) is added. The mixture is stirred at 25 °C for 8 h and concentrated in vacuo . The residue is taken up in diethyl ether and washed with saturated aqueous copper sulfate solution, water, saturated sodium chloride solution and dried over sodium sulfate. Concentration in vacuo gives 3.65 g of (2R,3S)-3-(formyl-2- tetrahydropyranyloxyamιno)-2-(4-phenylcyclohexylmethyl)butanoιc acid as a foam. The product is used without further purification.
Η NMR (300 MHz, DMSO-d6) δ 12.55 (br, IH), 8.43 and 8.20 (two m, IH), 7.31-7.15 (m, 5H), 4.92 (m, IH), 4.29 (m, IH), 3.95 (m, IH), 3.49 (m, IH), 2.66 (m, IH), 2.47 (m, 2H), 2.05-0.90 (m, 19H) ppm ESI-MS m z 402 (M-H) Example 2j; Pentafluorophenyl 3(2R,3S)-3-(Formyl-2-tetrahydropyranyloxyammo)-2-(4- phenylcyclohexylmethyl)butanoate
To a solution of (2R,3S)-3-(formyl-2-tefrahydropyranyloxyamιno)-2-(4- phenylcyclohexylmefhyl)butanoιc acid (3.65 g, 0.009 mol) in dichloromethane (20 mL) at 25 °C is added via syringe pyridme (0.87 g, 0.01 1 mol) followed by pentafluorophenyl tπfluoroacetate (3.1 g, 0.011 mol). The mixture is stirred for 6 h, diluted with dichloromethane and washed with 0.1 M aqueous HCl, 1 M aqueous sodium carbonate, saturated aqueous sodium chloride and dried over sodium sulfate. Concentration in vacuo followed by chromatography on silica gel with hexanes - EtOAc (2: 1) as eluent gives 3.05 g (60%o) of pentafluorophenyl (2R,3S)-3-(formyl-2-tefrahydropyranyloxyamιno)-2-(4- phenylcyclohexylmethyl)butanoate as an oil.
Η NMR (300 MHz, DMSO-d6) δ 8.49 and 8.30 (two m, IH), 7.32-7.16 (m, 5H), 4.97 (br s, IH), 4.48 and 4.22 (two m, IH), 3.97 (m, IH), 3.64 (m, IH), 3.32 (m, IH), 2.46 (m, IH), 2.12-0.99 (m, 20H) ppm.
Anal. Calcd. for C29H32F5N05: C, 61.15; H, 5.66; N, 2.46. Found: C, 61.23; H, 5.71; N, 2.40.
Example 2k; (2S)-2-tert-Butoxycarbonylammo-3,3-dιmethylbutanoιc Acid l,3-Thιazol-2- ylamide
A solution of 10.0 g (76.2 mmol) of (2S)-2-ammo-3,3-dιmethylbutanoιc acid in 100 mL of THF and 50 mL of water is freated at 25 °C with 20 mL (100 mmol) of 5 N aqueous sodium hydroxide followed by 20 g (91.6 mmol) of di-tert-butyl dicarbonate. The mixture is stirred vigorously at 25 °C for 24 h. The mixture is chilled to 0 °C and is treated dropwise with saturated aqueous sodium bisulfate solution to adjust the reaction mixture to pH 2. The mixture is extracted with two 200 - mL portions of EtOAc. The combined organic phases are dried over magnesium sulfate and concenfrated in vacuo to afford 18.5 g of crude (2S -2-tert- butoxycarbonylamιno-3,3-dιmethylbutanoιc acid. The crude acid is stirred in 200 mL of DMF at 0 °C as 12.3 g (91.1 mmol) of HOBt, 30 mL (273 mmol) of NMM, and 17.5 g (91.2 mmol) of EDC are added in turn After 30 mm at 0 °C 9.1 g (90.1 mmol) of 2-ammo-l,3- thiazole is added. The mixture is stirred at 0 °C for 30 min and at 50 °C for 2 h The mixture is then concentrated in vacuo and the residue is diluted with 250 mL of EtOAc. The organic phase is washed with water, saturated aqueous sodium chloride, is dried over magnesium sulfate, and concenfrated in vacuo. Chromatography on silica gel (elution with 30% EtOAc - hexanes followed by 70% EtOAc - hexanes) affords 20.1 g (84%) of (2S)-2-tert- butoxycarbonylammo-3,3-dιmethylbutanoιc acid l,3-fhιazol-2-ylamιde as an oil. Η NMR (300 MHz, CDC13) δ 7.52 (d, IH), 7.03 (d, IH), 5.42 (bd, IH), 3.41 (s, IH), 1.43 (s, 9H), 1.12 (s, 9H) ppm.
Example 21; (2S)-2-Ammo-3, 3 -dimethylbutanoic Acid l,3-Thιazol-2-ylamιde
A solution of 18.3 g (58.4 mmol) of (2S)-2-tert-butoxycarbonylamιno-3,3- dimethylbutanoic acid l,3-thιazol-2-ylamιde in 50 mL of dichloromethane is treated dropwise at 25 °C with 50 mL of TFA. After 4 h at 25 °C the mixture is concentrated in vacuo and the residue is diluted with 20 mL of dichloromethane. The mixture is stirred at 0 °C as saturated aqueous sodium carbonate is added dropwise to bring the mixture to pH 8. The mixture is diluted with water to a volume of 200 mL and the solid product is collected and dried in vacuo affording 11.6 g (93%) of (2S)-2-amιno-3,3-dιmethylbutanoιc acid 1 ,3-thιazol-2- ylamide.
Η NMR (300 MHz, CDC13) δ 7.50 (d, IH), 7.01 (d, IH), 3.42 (s, IH), 1.62 (bs, 2H), 1.11 (s, 9H) ppm.
Example 2m; (2R,3S)-3-(Formyl-tetrahydropyranyloxyamιno)-2-(4- phenylcyclohexylmethyl)butanoιc Acid [(lS)-2,2-Dιmethyl-l-(l,3-thιazol-2-ylcarbamoyl)-l- propyl] amide A mixture of pentafluorophenyl (2R,3S)-3-(formyl-2-tetrahydropyranyloxyammo)-2-(4- phenylcyclohexylmethyl)butanoate (0.25 g, 0.44 mmol), (2S)-2-ammo-3, 3 -dimethylbutanoic acid l ,3-thιazol-2-ylamιde (0.10 g, 0.49 mmol), HOBt (6.1 mg, 0.040 mmol) and TEA (0.07 g, 0.67 mmol) in DMF (3 mL) is heated at 41°C for 18 h. The reaction mixture is poured into a mixture of hexanes (100 mL) and EtOAc (100 mL) and the resulting mixture is washed with water, 1 M aqueous sodium carbonate and water. The organic phase is dried (sodium sulfate) and concentrated in vacuo and the residue is purified by column chromatography on silica gel with EtOAc - hexanes (1 : 1) as eluent to give 0.12 g (46%) of (2R,3S)-3-(formyl- tefrahydropyranyloxyamιno)-2-(4-phenylcyclohexylmethyl)butanoιc acid [(lS)-2,2-dιmethyl- l-(l,3-thιazol-2-ylcarbamoyl)-l-propyl]amιde as a white solid.
'H NMR (400 MHz, DMSO-d6) δ 12.36 (m, IH), 8.40 (m, IH), 8.32 and 8.09 (two s, IH), 7.50 (m, IH), 7.23-7.06 (m, 4H), 6.80 (m, 2H), 4.91 and 4.79 (two m, IH), 4.27 and 4.12 (two m, IH), 3.89 and 3.78 (two m, IH), 3.53 (m, IH), 3.02 (m, IH), 2.27 (m, IH), 2.05 (m, IH), 1.86-1.22 (m, 10H), 1.00 (m, 6H), 0.95 (s, 9H), 0.91 (m, IH), 0.74 (m, 2H) ppm. ESI-MS m z 597 (M-H)" .
Example 2; (2R,3S)-3-(Formyl-hydroxyammo)-2-(4-phenylcyclohexylmethyl)butanoιc Acid [(lS)-2,2-Dιmethyl-l-(l,3-thιazol-2-ylcarbamoyl)-l-propyl]amιde
A mixture of (2R,3S)-3-(formyl-tetrahydropyranyloxyamιno)-2-(4- phenylcyclohexylmethyl)butanoιc acid [( lS)-2,2-dιmethyl- 1 -( 1 ,3-thιazol-2-ylcarbamoyl)- 1 - propyl]amιde (0.12 g, 0.20 mmol) in 80% acetic acid (10 mL) is heated at 40 °C for 18 h. The mixture is concentrated in vauco, the residue is taken up in ethanol and concentrated in vacuo. Repeating the ethanol treatment several times and addition of water is followed b\ stirring at 25 °C for 18 h. The resulting solid is filtered, washed with water, and dried to give
0.083 g (80%) of (2R,3S)-3-(formyl-hydroxyamιno)-2-(4-phenylcyclohexylmethyl)butanoιc acid [(lS)-2,2-dιmethyl-l-(l,3-thιazol-2-ylcarbamoyl)-l-propyl]amιde as a white solid, m.p.
213-214 °C.
Η NMR (300 MHz, DMSO-d6) δ 12.33 (s, IH), 9.82 and 9.45 (two s, IH), 8.35 and 8.02 (two s, IH), 7.54 (d, IH), 7.19 (m, 4H), 6.92 (d, 2H), 4.76 (d, IH), 4.34 and 3.76 (two m, IH), 2.98 (m, IH), 2.32 (m, IH), 2.06 (m, IH), 1.72-0.76 (m, 22H) ppm. ESI-MS m/z 513 (M-H)".
Anal. Calcd. for C^N S. 0.3 C2H402: C, 62.23; H, 7 42; N, 10.52. Found: C, 62.44; H, 7.46; N, 10.32.
Example 3; (2R,3S)-3-(Formyl-hydroxyamιno)-2-(3-phenyl-l-propyl)hexanoιc Acid [(lS)-5- Benzyloxycarbonylamιno-l-(l,3,4-thιadιazol-2-yl-carbamoyl)-l-pentyllamιde
Figure imgf000133_0001
Example 3a; Ethyl (3R)-3-Hydroxyhexanoate and Methyl (3R)-3-Hydroxyhexanoate
Ethyl butyrylacetate (50.0 g, 316 mmol) is stirred m 75 mL of absolute ethanol as
[RuCl2(BINAP)]2 «NEt3 (0.139 g, 0.158 mmol) is added along with 2 N hydrochloric acid (0.158 mL, 0.316 mmol). The mixture is placed on a pressure hydrogenation apparatus and degassed by evacuating and filling with nitrogen several times. The vessel is then pressurized with hydrogen to 65 psi. The reaction is heated to 70 °C for 36 h and then is allowed to cool to 25 °C. The resulting reddish brown solution is concentrated under reduced pressure and the product distilled (40-50 °C, 200 milhtorr) to give a clear oil (50.0 g, 99% yield, >99% enantiomeπc excess determined by chiral analytical HPLC). Η NMR (400 MHz, CDC13) δ 4.17 (q, 2H), 4.01 (m, IH), 2.95 (d, IH), 2.47 (dd, IH), 2.40
(dd, IH), 1.58-1.38 (m, 4H), 1.38 (t, 3H), 0.94 (t, 3H) ppm.
Methyl (3R)-3-hydroxyhexanoate is prepared m the same manner described above in methanol employing methyl butyrylacetate as the starting ketoester. The enantiomenc excess is 99% as determined by chiral analytical HPLC methods.
Η NMR (300 MHz, CDC13) δ 4.04 (m, IH), 3.72 (s, 3H), 2.87 (d, IH), 2.50 (dd, IH), 2.46 (dd, IH), 1.58-1.38 (m, 4H), 0.94 (t, 3H) ppm.
Example 3b; Ethyl (2R,3R)-2-[(2£')-3-Phenyl-2-propen-l-yl]-3-hydroxyhexanoate
To a solution of dnsopropylamme (2.28 g, 20.63 mmol) in THF (20 mL) cooled to 0 °C is added n-butylhthium (20.63 mmol, 2.5 M in hexanes) and the resulting solution is stirred at 0 °C for 0.5 h. The reaction mixture is cooled to -50 °C followed by slow addition of ethyl (3R)-3-hydroxyhexanoate (1.5 g, 9.38 mmol). After 0.5 h a solution of cmnamyl bromide (2.67 g, 14.06 mmol) in HMPA (2 mL) is added and the reaction mixture is allowed to warm to -20 °C and stirred for 16 h. The reaction mixture is quenched by addition 3 mL of saturated aqueous ammonium chloride solution, is poured into 20 mL of 1 N hydrochloric acid, and is extracted with two 50 - mL portions of EtOAc. The combined organic layers are dried over magnesium sulfate, concentrated in vacuo, and purified by silica gel chromatography (elution with 25% EtOAc - hexanes) to afford ethyl (2R,3R)-2-[(2E)-3- phenyl-2-propen-l-yl]-3-hydroxyhexanoate as a yellow oil (1.82 g, 70% yield).
Η NMR (300 MHz, CDCI3) δ 7.42-7.22 (m, 5H), 6.52 (d, IH), 6.20 (dt, IH), 4.22 (q, 2H),
3.78 (m, IH), 2.64 (m, 4H), 1.54 (m, 2H), 1.26 (t, 3H), 0.96 (t, 3H) ppm. APCI-MS m/z 299 (M+Na)+.
Example 3c; Ethyl (2R,3R)-2-(3-Phenyl-l-propyl)-3-hydroxyhexanoate A solution of ethyl (2R,3R)-2-[(2£)-3-phenyl-2-propen-l-yl]-3- hydroxyhexanoate (1.76 g, 6.38 mmol) in 30 mL of EtOAc is freated with 200 mg of 10% palladium on carbon. The resulting suspension is repeatedly evacuated and purged with a hydrogen balloon, then stirred under 1 atmosphere pressure of hydrogen gas for 6 h The catalyst is filtered and the filtrate is concentrated in vacuo to provide ethyl (2R,3R)-2-(3- phenyl-l-propyl)-3-hydroxyhexanoate as an oil (1.70 g, 96% yield). 'H NMR (300 MHz, CDCI3) δ 7.42-7.20 (m, 5H), 4.22 (q, 2H), 3 70 (m, IH), 2.64 (m, 2H),
2.52 (m. 2H), 1.82 (m, IH), 1 70-1 40 (m, 6H), 1.26 (t, 3H), 0.96 (t, 3H) ppm. APCI-MS m/z 301 (M+Na)+.
Example 3d; (2R,3R)-2-(3-Phenyl-l-propyl)-3-hydroxyhexanoιc Acid
To a solution of ethyl (2R,3R)-2-(3-phenyl-l-propyl)-3-hydroxyhexanoate
(1.70 g, 6.12 mmol) in THF - ethanol (2:1, 18 mL) is added 1 N aqueous sodium hydroxide solution (6.7 mL, 6.7 mmol). The solution is stirred at 23 °C for 20 h and is acidified to pH 3 with saturated aqueous sodium bisulfate and is extracted with two 50 - mL portions of EtOAc. The combined organic layers are dried over anhydrous magnesium sulfate and concenfrated under reduced pressure to provide (2R,3R)-2-(3 -phenyl- l-propyl)-3-hydroxyhexanoιc acid as an oil (1.53 g, 100% yield). Η NMR (300 MHz, CDCI3) δ 7.40-7.20 (m, 5H), 3.78 (m, IH), 2.74 (m, 2H), 2.56 (m, IH),
1.94-1.64 (m, 4H), 1.58-1.40 (m, 4H), 0.96 (t, 3H) ppm. APCI-MS m/z 273 (M+Na)+.
Example 3e; (2R,3R)-2-(3-Phenyl-l-propyl)-3-hydroxyhexanoιc Acid 2- Tetrahydropyranyloxyamide To a solution of (2R,3R)-2-(3-phenyl-l-propyl)-3-hydroxyhexanoιc acid (1.53 g, 6.12 mmol) in dichloromethane (6 mL) is added 2-tetrahydropyranyloxyamme (0.93 g, 7.96 mmol) and EDC (1.29 g, 6.73 mmol). The resulting solution is stirred at 23 °C for 3 h, concenfrated in vacuo, and diluted with 50 mL of EtOAc. The organic layer is washed sequentially with 1 N hydrochloric acid, saturated aqueous sodium bicarbonate solution, saturated aqueous sodium chloride solution, and is dried over anhydrous magnesium sulfate. Concentration under reduced pressure and purification by silica gel chromatography (elution with 2: 1 EtOAc - hexanes) provides (2R,3R)-2-(3 -phenyl- l-propyl)-3- hydroxyhexanoic acid 2-tetrahydropyranyloxyamιde as a solid (1.62 g, 76% yield).
Η NMR (400 MHz, CDCI3) δ 7.25 (m, 2H), 7.18 (m, 3H), 4.95 (m, IH), 3.93 (m, IH), 3.62
(m, 2H), 2.80 (d, IH), 2.62 (t, 2H), 2.04 (m, IH), 1.84 (m, 4H), 1.66-1.54 (m, 6H), 1.46-1.30 (m, 4H), 0.96 (t, 3H) ppm. APCI-MS m z 372 (M+Na)+.
Example 3 f; (3R,4S)- 1 -(2-Tetrahydropyranyloxy)-3-(3 -phenyl- 1 -propyl)-4-propylazetιdm-2- one
To a solution of (2R,3R)-2-(3-phenyl-l-propyl)-3-hydroxyhexanoιc acid 2- tefrahydropyranyloxyamide (1.62 g, 4.64 mmol) in 7 mL of dichloromethane at 0 °C is added pyridme (1 mL) and methanesulfonyl chloride (0.58 g, 5.10 mmol). The resulting solution is allowed to warm to 23 °C and is stirred at 23 °C for 24 h, concentrated in vacuo, and diluted with EtOAc (40 mL). The organic layer is washed with 1 N hydrochloric acid, saturated aqueous cupric sulfate solution, dried over anhydrous magnesium sulfate, and concenfrated to provide the desired methanesulfonate intermediate.
A suspension of potassium carbonate (1.92 g) in acetone (90 mL) is heated to reflux for 1 h. A solution of the above methanesulfonate in acetone (10 mL) is added and the resulting suspension is heated at reflux for 1 h. The mixture is allowed to cool to 25 °C and is filtered, concentrated under reduced pressure, and purified by silica gel chromatography (elution with 2: 1 hexanes - EtOAc) to provide (3R,4S)-l-(2-tetrahydropyranyloxy)-3-(3- phenyl-l-propyl)-4-propylazetιdm-2-one as an oil (1.22 g, 79% yield). 'H NMR (300 MHz, CDCI3) δ 7.25 (m, 2H), 7.18 (m, 3H), 5.20 and 5.02 (two m, IH), 4.30-
4.08 (m, IH), 3.92 (m, IH), 3.62 (m, IH), 2.98 (m, IH), 2.68 (m, 2H), 1.96 (m, IH), 1.84- 1.32 (m, 13H), 0.96 (m, 3H) ppm. APCI-MS m/z 354 (M+Na)+.
Example 3g; (2R,3S)-2-(3-Phenyl-l-propyl)-3-(2-tetrahydropyranyloxyamιno)hexanoιc Acid
To a solution of (3R,4S)-l-(2-tetrahydropyranyloxy)-3-(3-phenyl-l-propyl)-4- propylazetιdm-2-one (1.22 g, 3.69 mmol) in THF - methanol (2: 1, 15 mL) is added 1 N aqueous sodium hydroxide (5.5 mL). The solution is stirred at 23 °C for 36 h, is acidified to pH 3 with saturated aqueous sodium bisulfate solution, and is extracted with two 100 - mL portions of EtOAc. The combined organics are dried over anhydrous sodium sulfate and concentrated under reduced pressure to provide (2R,3S)-2-(3-phenyl-l-propyl)-3-(2- tetrahydropyranyloxyamιno)hexanoιc acid as an oil (0.93 g, 72% yield). Η NMR (300 MHz, CDCI3) δ 7.34 (m, 2H), 7.21 (m, 3H), 4.84 and 4.75 (two m, IH), 4.06
and 3.94 (two m, IH), 3.62 (m, IH), 3.20 and 3.10 (two m, IH), 3.01 and 2.92 (two m, IH), 2.62 (m, IH), 1.96-1.21 (m, 14H), 0.96 (m, 3H) ppm. APCI-MS m z 350 (M+H)+.
Example 3h; Pentafluorophenyl (2R,3S)-2-(3-Phenyl-l-propyl)-3-(formyl-2- tefrahydropyranyloxyamιno)hexanoate To a solution of (2R,3S)-2-(3-phenyl-l-propyl)-3-(2- tefrahydropyranyloxyamιno)hexanoιc acid (0.93 g, 2.66 mmol) m pyridine (2 mL) at 0 °C is added formic acetic anhydride ( 0 4 mL) The resulting solution is stirred at 0 °C for 1 h, concenfrated in vacuo, and diluted with 1 1 mL of EtOAc. To the solution of crude acid is added pentafluorophenol (0.51 g, 2.79 mmol), NMM (0.28 g, 2.79 mmol) and dicyclohexylcarbodiimide (0.58 g, 2.79 mmol). The resulting solution is stirred at 23 °C for 20 h and is filtered. The filtrate is washed with 1 N hydrochloric acid, 1 M aqueous sodium carbonate solution, and saturated aqueous sodium chloride solution. The organic layer is dried over magnesium sulfate, concenfrated, and purified by silica gel chromatography (elution with 9: 1 hexanes - EtOAc) to provide pentafluorophenyl (2R,3S)-2-(3-phenyl-l- propyl)-3-(formyl-2-tetrahydropyranyloxyamιno)hexanoate as an oil (0.84 g, 58% yield). Η NMR (400 MHz, CDCI3) δ 8.56 and 8.02 (two s, IH), 7.28-7.14 (m, 5H), 4.76 and 4.72
(two m, IH), 4.56 and 3.68 (m, IH), 3.96 (m, IH), 3.56 (m, IH), 3.18 and 3.02 (two m, IH), 2.64 (m, 2H), 2.02-1.38 (m, 14H), 0.90 (m, 3H) ppm APCI-MS m z 566 (M+Na)+.
Example 3ι; (2S)-6-Benzyloxycarbonylamιno-2-tert-butoxycarbonylammohexanoιc Acid 1 ,3,4-Thιadιazol-2-ylamιde
To a solution of (2S)-6-benzyloxycarbonylamιno-2-tert- butoxycarbonylammohexanoic acid (1.09 g, 2.87 mmol) in dichloromethane (5 mL) is added 1 , 1 -carbonyldiimidazole (0.47 g, 2.87 mmol). The resulting solution is stirred at 25 °C for 1 h and 2-ammo- 1,3, 4- thiadiazole (0.29 g, 2 87 mmol) is added and the reaction is stirred for an additional 18 h. The mixture is diluted with dichloromethane (60 mL) and washed with 1 M aqueous sodium carbonate solution. The organic layer is dπed over anhydrous magnesium sulfate, concentrated in vacuo, and purified by silica gel chromatography (elution with 1:1 EtOAc - hexanes) to provide (2S)-6-benzyloxycarbonylammo-2-tert- butoxycarbonylaminohexanoic acid l,3,4-thιadιazol-2-ylamιde as a foam (0.81 g, 61% yield).
Η NMR (300 MHz, CDCI3) δ 13.35 (bs, IH), 8.78 (bs, IH), 7.32 (m, 5H), 6.56 (m, IH), 5.09
(m, 3H), 4.46 (m, IH), 3.21 (m, 2H), 2.91 (m, 2H), 1.95-1.56 (m, 4H), 1.26 (s, 9H) ppm. APCI-MS m/z 464 (M+H)+.
Example 3j; (2S)-6-Benzyloxycarbonylammo-2-amιnohexanoιc Acid l,3,4-Thιadιazol-2- ylamide
To a solution of (2S)-6-benzyloxycarbonylammo-2-tert- butoxycarbonylammohexanoic acid l,3,4-thιadιazol-2-ylamιde (0.81 g, 1.75 mmol) in dichloromethane (8 mL) is added frifluoroacetic acid (2 mL) The resulting solution is stirred for 4 h at 25 °C, is concenfrated, diluted with 50 mL of EtOAc, and washed with 1 N aqueous sodium hydroxide solution. The organic layer is dried over anhydrous sodium sulfate and concentrated under reduced pressure to provide (2S)-6-benzyloxycarbonylammo- 2-ammohexanoιc acid l,3,4-thιadιazol-2-ylamιde as a solid (0.62 g, 98% yield) Η NMR (300 MHz, CDCI3) δ 8.82 (s, IH), 7.31 (m, 5H), 5 42 (bs, IH), 5.04 (s, 2H), 3.64
(m, IH), 3.16 (m, 4H), 1.85 (m, IH), 1.62 (m, IH), 1.51 (m, 4H) ppm. APCI-MS m/z 364 (M+H)+.
Example 3k; (2R,3S)-3-(Formyl-2-tetrahydropyranyloxyamιno)-2-(3-phenyl-l- propyl)hexanoιc Acid [( 1 S)-5-Benzyloxycarbonylamιno- 1 -( 1 ,3 ,4-fhιadιazol-2-ylcarbamoyl)- 1 -pentyl] amide To a solution of pentafluorophenyl (2R,3S)-2-(3-phenyl-l-propyl)-3-(formyl-2- tetrahydropyranyloxyamιno)hexanoate (90 mg, 0.166 mmol) in DMF (0.5 mL) is added (2S)- 6-benzyloxycarbonylamιno-2-ammohexanoιc acid l,3,4-thιadιazol-2-ylamιde (78 mg, 0.215 mmol) and HOBt ( 2.2 mg, 0.017 mmol). The resulting solution is heated to 50 °C and is stirred for 18 h. The reaction mixture is cooled to 25 °C, concentrated in vacuo, diluted with 20 mL of EtOAc, and washed with 1 M aqueous sodium carbonate solution. The organic layer is dried over magnesium sulfate, concentrated in vacuo, and purified by silica gel chromatography (elution with 2: 1 EtOAc - hexane) to provide (2R,3S)-3-(formyl-2- tefrahydropyranyloxyamιno)-2-(3-phenyl-l-propyl)hexanoιc acid [(lS)-5- benzyloxycarbonylammo-l-(l,3,4-thιadιazol-2-ylcarbamoyl)-l-pentyl]amιde as an oil (24.1 mg, 25% yield). APCI-MS m/z 745 (M+Na)+.
Example 3; (2R,3S)-3-(Foπnyl-hydroxyammo)-2-(3-phenyl-l-propyl)hexanoιc Acid [(lS)-5- Benzyloxycarbonylamino- 1 -( 1 ,3 ,4-thιadιazol-2-ylcarbamoyl)- 1 -pentyljamide
A solution of (2R,3S)-3-(formyl-2-tetrahydropyranyloxyammo)-2-(3-phenyl-l- propyl)hexanoιc acid [( 1 S)-5-benzyloxycarbonylammo- 1 -( 1 ,3 ,4-thιadιazol-2-ylcarbamoyl)- 1 - pentyl]amιde (23 mg, 0.032 mmol) in acetic acid - water (4: 1, 1 mL) is heated to 50 °C for 16 h. The reaction mixture is concenfrated, then dissolved in toluene and concentrated in vacuo. The procedure is repeated once again to afford the crude product which is recrystallized from dichloromethane - methanol - diethyl ether to provide (2R,3S)-3-(formyl-hydroxyamιno)-2- (3 -phenyl- 1 -propyl)hexanoιc acid [( 1 S)-5-benzyloxycarbonylammo- 1 -( 1 ,3 ,4-thιadιazol-2- ylcarbamoyl)-l-pentyl]amιde as a solid (14 mg, 69% yield). Η NMR (400 MHz, CD3OD) δ 9.04 (s, IH), 8.35 and 7.95 (two s, IH), 7.30 (m, 5H), 7.12
(m, 5H), 5.03 (m, 2H), 4.57 (m, IH), 4.39 and 3.61 (two dt, IH), 3.11 (t, 2H), 2.78-2.46 (m,
2H), 1.78-1.14 (m, 14H), 0.88 (m, 3H) ppm.
APCI-MS m z 585 (M+Na)+.
Anal. Calcd. for C26H38N606S 0.5 H20: C, 54.62; H, 6.87; N, 14.70. Found: C, 54.70; H,
6.72; N, 14.41.
Example 195; (2R,3S)-3-(Formylhydroxyamιno)-2-(3-(4-chlorophenyl)-l-propyl)butanoιc Acid ["( 1 S)-2,2-Dιmethyl-( 1 -(3 -pyπdyl)carbamoyl)- 1 -propyl]amιde
Figure imgf000141_0001
Example 195a; Methyl (2R,3R)-2-(3-tπmethylsιlyl-2-propyne-l-yl)-3-hydroxybutanoate
To a solution of dnsopropylamine (9.42 g, 93.22 mmol) in THF (100 mL) cooled to -
50 °C is added n-butylhthium (93.22 mmol, 2.5M in hexanes) and the resulting solution is stirred at -50 °C for 0.5 h. The reaction mixture is cooled to -78 °C followed by slow addition of methyl (3R)-3-hydroxybutanoate (5 g, 42.37 mmol). After 0.5 h a solution of 3- tπmethylsilylpropargyl bromide (9.76 g, 50.85 mmol) in HMPA (1 mL) is added and the reaction mixture is allowed to warm to 0 °C and stirred for 16 h. The reaction mixture is quenched by addition 10 mL of saturated aqueous ammonium chloride solution, is poured into 100 mL of 1 M hydrochloric acid, and is extracted with two 100 mL portions of EtOAc. The combined organic layers are dried over magnesium sulfate, concentrated in vacuo, and purified by silica gel chromatography (elution with 20% EtOAc - hexanes) to afford methyl
(2R,3R)-2-(3-tπmethylsιlyl-2-propyne-l-yl)-3-hydroxybutanoate as a yellow oil (5.6 g, 58% yield).
Η NMR (400 MHz, CDCI3) δ 4.04 (m, IH), 3.72 (s, 3H), 2.62 (m, 3H), 1.23 (t, 3H), 0.09 (s,
9H) ppm.
ESI-MS m z 251.2 (M+Na)+.
Example 195b; (2R,3R)-2-(2-propyne-l-yl)-3-hydroxybutanoιc Acid
To a solution of methyl (2R,3R)-2-(3-tπmethylsιlyl-2-propyne-l-yl)-3- hydroxybutanoate (5.6 g, 24.56 mmol) in THF - methanol (3.1, 160 mL) is added 2 M aqueous sodium hydroxide solution (40 mL, 36.8 mmol). The solution is stirred at 23 °C for 20 h, then concentrated and exfracted with hexanes (100 mL). The aqueous layer is acidified to pH 3 with 1 M HCl and is extracted with two 100 mL portions of EtOAc. The combined organic layers are dried over anhydrous magnesium sulfate and concenfrated under reduced pressure to provide (2R,3R)-2-(2-propyne-l-yl)-3-hydroxybutanoιc acid as an oil (3.0 g, 86% yield). Η NMR (400 MHz, CDCI3) δ 7.28 (m, 2H), 7.16 (m, 3H), 3.93(m, IH), 2.63 (m, 2H), 2.43
(m, IH), 1.69 (m, 4H), 1.26 (d, 3H) ppm. ESI-MS m z 221.3 (M-H)".
Example 195c; (2R,3R)-2-(2-propyne-l-yl)-3-hydroxybutanoιc Acid 2- Tetrahydropyranyloxyamide
To a solution of (2R,3R)-2-(2-propyne-l-yl)-3-hydroxybutanoιc acid (3.0 g, 21.13 mmol) in dichloromethane (50 mL) is added 2-tetrahydropyranyloxyamιne (3.0 g, 25.35 mmol) and EDC (4.5 g, 23.24 mmol). The resulting solution is stirred at 23 °C for 4 h, then diluted with dichloromethane (100 mL) and washed with 1 M hydrochloric acid. The combined organic layers were dried over anhydrous magnesium sulfate and concentrated under reduced pressure to provide (2R,3R)-2-(2-propyne-l-yl)-3-hydroxybutanoic acid 2- tetrahydropyranyloxyamide as a foam (1.9 g, 37% yield). 'H NMR (400 MHz, CDCI3) δ
8.82 (bs, IH), 4.96 (m, IH), 3.94 (m, 2H), 3.62 (m, IH), 2.64 (m, IH), 2.56 (m, IH), 2.23 (m, IH), 2.04 (m, IH), 1.82 (m, 2H), 1.61 (m, 4H), 1.23 (d, 3H) ppm. ESI-MS m/z 240.4 (M-H)".
Example 195d; (3R,4S)- 1 -(2-Tefrahydropyranyloxy)-3-(2-propyne- 1 -yl)-4-methylazetidin-2- one
To a solution of (2R,3R)-2-(2-propyne-l-yl)-3-hydroxybutanoic acid 2- tetrahydropyranyloxyamide (1.9 g, 7.88 mmol) in 20 mL of dichloromethane at 0 °C is added pyridine (5 mL) and methanesulfonyl chloride (0.99 g, 8.67 mmol). The resulting solution is allowed to warm to 23 °C and is stirred at 23 °C for 16 h, concentrated in vacuo, and diluted with dichloromethane (100 mL). The organic layer is washed with 1 M hydrochloric acid, saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, and concentrated to provide the desired methanesulfonate intermediate.
A suspension of potassium carbonate (3.26 g) in acetone (10 mL) is heated to reflux for 1 h. A solution of the above methanesulfonate in acetone (100 mL) is added and the resulting suspension is heated at reflux for 6 h. The mixture is allowed to cool to 25 °C and is filtered, concentrated under reduced pressure, and purified by silica gel chromatography (elution with 4: 1 hexanes - EtOAc) to provide (3R,4S)-l-(2-tetrahydropyranyloxy)-3-(2- propyne-l-yl)-4-methylazetidin-2-one as an oil (1.3 g, 74% yield). Η NMR (400 MHz, CDCI3) δ 5.15 (m, 0.5H), 4.99 (m, 0.5H), 4.20-4.01 (m, 2H), 3.62 (m,
IH), 3.18 (m, IH), 2.63 (m, IH), 2.40 (m, IH), 1.99 (m, IH), 1.89-1.51 (m, 6H), 1.42 (d, 1.5H), 1.38 (d, 1.5H) ppm. ESI-MS m/z 246.3 (M+Na)+
Example 195e; (3R,4S)-l-(2-Tetrahydropyranyloxy)-3-((2E)-3-tπbutylstannyl-2-propene-l- yl)-4-methylazetιdιn-2-one
To a solution of (3R,4S)-l-(2-tetrahydropyranyloxy)-3-(2-propyne-l-yl)-4- methylazetιdιn-2-one (1.32 g, 5 83 mmol) m 20 mL of toluene is added tπbutyltm hydride (1 70 g, 5.83 mmol) and AJBN (30 mg). The resulting solution is heated at reflux for 4 h then concenfrated in vacuo. The reaction mixture is purified by silica gel chromatography (elution with 8: 1 hexanes - EtOAc) to provide (3R,4S)-l-(2-tefrahydropyranyloxy)-3-((2E)-3- tπbutylstannyl-2-propene-l-yl)-4-methylazetιdιn-2-one as an oil (2.6 g, 87% yield) Η NMR (400 MHz, CDCI3) δ 5.94 (m, 2H), 5.16 (m, 0.5H), 4.99 (m, 0.5H), 4.20-4.01 (m,
2H), 3.62 (m, IH), 3.12 (m, IH), 2.61 (m, IH), 2.36 (m, IH), 1.76 (m, 2H), 1.58 (m, 4H), 1.44 (m, 6H), 1.26 (m, 9H), 0.86 (m, 15H) ppm. ESI-MS m z 538.2 (M+Na)+.
Example 195f; (3R,4S)-l-(2-Tetrahydropyranyloxy)-3-((2E)-3-(4-chlorophenyl)-2-propene-l- yl)-4-methylazetιdιn-2-one
To a solution of (3R,4S)-l-(2-tetrahydropyranyloxy)-3-((2E)-3-tπbutylstannyl-2- propene-l-yl)-4-methylazetιdm-2-one (0.92 g, 1.79 mmol) in 3 mL of dimethylformamide is added 4-chloroιodobenzene (470 mg, 1.97 mmol) and tπphenyphosphme palladium (II) dichloride (63 mg, 0.09 mmol). The resulting solution is heated at 80°C for 16 h, then 0.5 mL ammonium hydroxide is added. The reaction mixture is poured into saturated sodium chloride solution (20 mL) and extracted with 1 : 1 EtOAc - hexane (50 mL). The combined organic layers were dπed over anhydrous magnesium sulfate, concentrated under reduced pressure, and purified by silica gel chromatography (elution with 3: 1 hexanes - EtOAc) to provide (3R,4S)- 1 -(2-tetrahydropyranyloxy)-3-((2E)-3-(4-chlorophenyl)-2-propene- 1 -yl)-4- methylazetιdιn-2-one as an oil (370 mg, 61% yield).
Η NMR (400 MHz, CDCI3) δ 7.30 (d, 2H), 7.16 (d, 2H), 6.38 (m, IH), 6.18 (m, IH), 5.16
(m, 0.5H), 4.99 (m, 0.5H), 4.20-4.01 (m, 2H), 3.62 (m, IH), 3.12 (m, IH), 2.62 (m, IH), 2.42 (m, IH), 1.76 (m, 2H), 1.58 (m, 4H), 1.36 (m, 3H) ppm. ESI-MS m/z 358.2 (M+Na)+.
Example 195g; (3R,4S)-l-(2-Tetrahydropyranyloxy)-3-( 3-(4-chlorophenyl)-l-propyl)-4- methylazetιdιn-2-one
To a solution of (3R,4S)-l-(2-tefrahydropyranyloxy)-3-((2E)-3-(4-chlorophenyl)-2- propene-l-yl)-4-methylazetιdιn-2-one (0.37 g, 1.10 mmol) in 5 mL of methanol is treated with 30 mg of 5% palladium on barium sulfate. The resulting suspension is repeatedly evacuated and purged with a hydrogen balloon, then stirred under 1 atmosphere pressure of
« hydrogen gas for 30 h. The catalyst is filtered and the filfrate is concentrated in vacuo to provide (3R,4S)-l-(2-tefrahydropyranyloxy)-3-(3-(4-chlorophenyl)-l-propyl)-4- methylazetιdιn-2-one as an oil (360 mg, 97% yield).
Η NMR (400 MHz, CDCI3) δ 7.22 (d, 2H), 7.08 (d, 2H), 5.16 (m, 0.5H), 4.96 (m, 0.5H),
4.16-3.96 (m, 2H), 3.62 (m, IH), 2.88 (m, IH), 2.62 ( , 2H), 1.82-1.44 (m, 10H), 1.22 (m, 3H) ppm.
ESI-MS m/z 360.3 (M+Na)+.
Example 195h; (2R,3S)-2-(3-(4-chlorophenyl)-l-propyl)-3-(2- tefrahydropyranyloxyamino)butanoic Acid
To a solution of (3R,4S)-l-(2-tefrahydropyranyloxy)-3-(3-(4-chlorophenyl)-l- propyl)-4-mefhylazetιdιn-2-one (360 mg, 1.07 mmol) in dioxane (3.2 mL) is added 1 M aqueous sodium hydroxide (1.6 mL). The solution is stirred at 23 °C for 72 h, then extracted with hexanes (20 mL). The aqueous layer is acidified to pH 3 with saturated aqueous sodium bisulfate solution, and is extracted with two 30 mL portions of EtOAc. The combined organics are washed with saturated aqueous sodium chloride, dπed over anhydrous magnesium sulfate, and concenfrated under reduced pressure to provide (2R,3S)-2-(3-(4- chlorophenyl)-l-propyl)-3-(2-tefrahydropyranyloxyammo)butanoιc acid as an oil (380 mg, 99% yield). Η NMR (400 MHz, CDCI3) δ 7.22 (d, 2H), 7.06 (d, 2H), 4.92 (m, 0.5H), 4.78 (m, 0.5H),
3.95 (m. 0.5H), 3.86 (m, 0.5H), 3.57 (m, IH), 3.36 (m, 0.5H), 3.24 (m, 0.5H), 2.94 (m, 0.5H), 2.81 (m, 0.5H), 2.62 (m, 2H), 1.94-1.66 (m, 4H), 1.62-1.44 (m, 6H), 1.24 (m, IH), 1.08 (d, 1.5H), 1.02 (d, 1.5H) ppm. ESI-MS m/z 354.2 (M-H)".
Example 195ι; (2R,3S)-2-(3-(4-chlorophenyl)-l-propyl)-3-(formyl-2- tefrahydropyranyloxyamιno)butanoιc Acid
To a solution of (2R,3S)-2-(3-(4-chlorophenyl)-l-propyl)-3-(2- tetrahydropyranyloxyammo)butanoιc acid (380 mg, 1.07 mmol) in pyridme (4 mL) at 0 °C is added formic acetic anhydride ( 0.9 mL). The resulting solution is allowed to warm to 25 °C, stirred for 3 h, and then concentrated to dryness under reduced pressure. The resulting gum is dissolved in EtOAc (30 mL) and washed sequentially with 1 M hydrochloric acid (20 mL) and saturated aqueous sodium chloride solution. The organic layer is dried over anhydrous magnesium sulfate and concenfrated under reduced pressure to provide (2R,3S)-2-(3-(4- chlorophenyl)-l-propyl)-3-(formyl-2-tetrahydropyranyloxyamιno)butanoιc acid as an oil (385 mg, 94% yield).
ESI-MS m/z 406.2 (TVI+Naf, 382.3 (M-H)". Example 195j; (2R,3S)-3-(Formyl-2-tetrahydropyranyloxyamιno)-2-(3-(4-chlorophenyl)-l- propyl)butanoιc Acid [( 1 S)-2,2-Dιmethyl-( 1 -(3 -pyπdyl)carbamoyl)- 1 -propy ljamide
To a solution of (2R,3S)-2-(3-(4-chlorophenyl)-l-propyl)-3-(formyl-2- tefrahydropyranyloxyamιno)butanoιc acid (400 mg, 1.04 mmol) in DMF (4 mL) is added BOP reagent (507 mg, 1.15 mmol), HOBt ( 155 mg, 1.15 mmol), and NMM (527 mg, 5.22 mmol). After 30 mm, addition of (2S)-2-Ammo-3,3-dιmethylbutanoιc acid 3-pyπdylamιde hydrochloπde (447 mg, 1.56 mmol) occurs and the resulting solution is stirred at 25 °C for 72 h. The reaction mixture is poured into EtOAc - hexanes (1 : 1, 200 mL) and washed sequentially with 1 M aqueous sodium carbonate and saturated aqueous sodium chloride. The organic layer is dried over magnesium sulfate, concenfrated in vacuo, and purified by silica gel chromatography (elution with 3: 1 EtOAc - hexane) to provide (2R,3S)-3-(Formyl-2- tetrahydropyranyloxyamιno)-2-(3-(4-chlorophenyl)- 1 -propyl)butanoιc acid [( lS)-2,2- dιmethyl-(l-(3-pyπdyl)carbamoyl)-l-propyl]amιde as a white solid (441 mg, 74% yield). ESI-MS m/z 595.1 (M+Na)\
Example 195; (2R,3S)-3-(Formylhydroxyamιno)-2-(3-(4-chlorophenyl)-l-propyl)butanoιc acid [(lS)-2,2-Dιmethyl- 1 -(3-pyπdyl)carbamoyl)- 1 -propyljamide
A solution of (2R,3S)-3-(Formyl-2-tetrahydropyranyloxyammo)-2-(3-(4- chlorophenyl)- 1 -propyl)butanoιc acid [( 1 S)-2,2-dιmethyl-( 1 -(3-pyπdyl)carbamoyl)- 1 - propyljamide (440 mg, 0.77 mmol) in acetic acid - water (4: 1, 2 mL) is heated to 50 °C for 20 h. The reaction mixture is concenfrated, then dissolved in toluene and concenfrated in vacuo. The procedure is repeated once again to afford the crude product which is triturated from hot dichloromethane - diethyl ether to provide (2R,3S)-3-(formylhydroxyammo)-2-(3-(4- chlorophenyl)- 1 -propyl)butanoιc acid [( lS)-2,2-dιmethyl-( 1 -(3-pyπdyl)carbamoyl)- 1 - propyljamide as a white solid (297 mg, 79% yield). 'H NMR (400 MHz, CD3OD) δ 8.75 (s, IH), 8.27 (m, IH), 8.25 and 7.98 (s, IH), 8 00 (m,
IH), 7.38 (m, IH), 6.97 (m, 4H), 4.48 and 3.82 (m, IH), 4.45 (m, IH), 2.94 and 2 84 (m, IH), 2.58 (m, lH), 2.39 (m, IH), 1.48 (m, 4H), 1.24 and 1.18 (d, 3H), 1.03 (s, 9H) ppm. ESI-MS m/z 511.3 (M+Na)+.
PHARMACOLOGY
The efficacy of compounds of the present invention as inhibitors of matnx metalloproteases, TNF converting enzyme and TNFα cellular release can be evaluated and measured using pharmacological methods known in the art or as described in detail below based on similarly established methodologies.
Pharmacological Example 1
A. Matrix Metalloprotease Inhibition Protocol
The potency of compounds of the invention as inhibitors of 19 kD truncated collagenase- 1 (MMP-1), 20 kD truncated collagenase-3 (MMP-13), stromelysm- 1 (MMP-3), and 50 kD truncated gelatinase B (MMP-9) is determined according to the general procedure of Bickett et. al. (Anal. Biochem. 1993, 212, 58-64) using the fluorogenic substrate, DNP-Pro- Cha-Gly-Cys(Me)-Hιs-Ala-Lys(NMA)-NH2 (DNP = 2,4-dmιtrophenyl, NMA = N-
methylanthranihc acid). Assays are conducted m a total volume of 0.180 mL assay buffer (200 mM NaCl, 50 mM Tπs, 5 mM CaCl2, 10 μM ZnS0 , 0.005% Bπj 35, pH 7.6) in each
well of a black 96 well microtiter plate. 19 kD collagenase- 1, 20 kD collagenase-3, sfromelysm-l, and 50 kD gelatinase B concenfrations are adjusted to 500 pM, 30 pM, 5 nM, and 100 pM, respectively. A dose response is generated using an eleven - point, 3 - fold seπal dilution with initial starting test compound concenfrations of 100, 10, or 1 μM. Inhibitor and enzyme reactions are incubated for 30 minutes at ambient temperature and then initiated with
10 μM fluorogenic substrate (above). The product formation is measured at Excιtatιon343/Emιssιon45o nm after 45-180 minutes using a Fluostar SLT fluorescence
analyzer. Percent inhibition is calculated at each inhibitor concentration and the data were plotted using standard curve fitting programs. IC50 values were determined from these curves. Assays were run at low substrate concentration ([S]«Km) such that the calculated IC5o values are equivalent to K, within experimental error.
B. TNFα Converting Enzyme Inhibition Protocol
The potency of compounds of the invention as inhibitors of cell - free tumor necrosis factor α converting enzyme is determined as follows; Membrane preparation from MonoMac 6 cells (subfractionated extract from equivalent of 6x106 cells per 60 μl assay) is incubated for 1 h with 200 nM radiolabeled substrate (Bιotm-SPLAQAVRSSSRT-(3H)P-S-NH2 , 4.1 Ci/mmol, ref # 0935 from Zeneca) in 10 mM hepes buffer, 250 mM sucrose, pH 7.5. The reaction is quenched by addition of streptavidin SPA beads (Amersham RPNQ0006), with excess binding capacity relative to substrate, suspended in 250 mM EDTA, pH 8.0. Binding is complete within 15 minutes and plates are counted m a Wallac 1450 Microbeta liquid scintillation counter. Percent inhibition is calculated at each inhibitor concentration and the data were plotted using standard curve fitting programs. IC50 values were determined from these curves. Assays were run at low substrate concentration ([S]«Km) such that the calculated IC50 values are equivalent to K, within experimental error.
C. Cell - Based TNFα Release Inhibition Protocol
The potency of compounds of the invention as inhibitors of release of soluble tumor necrosis factor α from stimulated monocytes in vitro is determined as follows; LPS/PMA solution for assay consisting of a) 4 μL of 5 mg/mL LPS stock and b) 6 μL of 10 mg/mL
PMA stock are added to 500 μL of medium (RPMI 1640 (Gibco) + 10% FBS + penicillin/streptomycin + 1-glutamιne). This solution is then diluted 1.1000 (40 ng/mL and 120 ng/mL) for use later in the assay. Compounds (10 mM) are serially diluted 1 :3 in DMSO. Compound dilutions (20 μL) are added to a sterile round bottom 96 well plate (20 μL:200 μL total volume = 1 : 10 for final concenfrations of 50 μM for test compounds). MonoMac 6 cell suspension (130 μL, 1.5 xlO6 cells/mL) is then added to each well resulting m 2 x 105 cells/well. LPS/PMA (50 μL) solution is then added to each well to begin stimulation (final concentrations of 10 ng/mL and 30 ng/mL respectively) The plate is incubated at 37 °C for 2 hours then spun at 1,500 rpm for 3 minutes to pellet cells. The supernatant (120 μL/well) is removed to a new round bottom 96 well plate and diluted 1 : 10 PBS. Then, 20 μL of the supernatant is transferred to a Cistron TNFα ELISA plate and processed according to the manufacturer's instructions to quantitate levels of TNFα. Percent inhibition of TNFα release is calculated at each inhibitor concentration and the data were plotted using standard curve fitting programs. IC50 values were determined from these curves.
Results are listed in Table 3.
Table 3
Figure imgf000151_0001
Pharmacological Example 2
Murine LPS - Stimulated Serum TNF Inhibition Protocol
The potency of compounds of the invention as inhibitors of serum TNFα elevation in mice treated with lipopolysaccharide (LPS) is determined as follows; a) for subcutaneous (s.c.) administration, test compound is dissolved in DMSO and added to a mixture of 0.9% sodium chloride solution and 30% Trappsol HPB-20 (Cyclodextrin Technology Development Inc., Gainesville, Florida USA) for a final DMSO concentration of 1%. The dosmg solution is sonicated briefly and 0.2 mL is injected subcutaneously 10 mm prior to LPS injection, b) for per oral (p.o.) administration, test compounds are formulated in 0.2 mL of PBS and 0.1% Tween 80 and given orally via gavage 10 mm prior to LPS administration. C3/hen female mice are injected mfrapeπtoneally with 200 μg/kg LPS (Escherichia coli, Serotype 0111 :B4, Sigma Chemical Co, St. Louis, MO) in PBS and sacrificed 90 mm later by C02 asphyxiation. Blood is immediately taken from the caudal vena cava and plasma prepared and frozen at -80 °C. Plasma concenfrations of TNF are measured by ELISA (Genzyme Co., Cambridge MA). Results are listed in Table 4.
Table 4
Figure imgf000153_0001
Throughout this application, various publications are may be recited. Such publications are hereby incorporated by reference m their entirety
While the invention has been described and illustrated with reference to certain preferred embodiments thereof, those skilled in the art will appreciate that various changes, modifications and substitutions can be made therein without departing from the spirit and scope of the invention. For example, effective dosages other than the preferred dosages as set forth herein above may be applicable as a consequence of variations in the responsiveness of the mammal being treated for inflammatory conditions, or for other indications for the compounds of the invention indicated above. Likewise, the specific pharmacological responses observed may vary according to and depending upon the particular active compound selected or whether there are present pharmaceutical carriers, as well as the type of formulation and mode of administration employed, and such expected variations or differences in the results are contemplated m accordance with the objects and practices of the present invention. It is intended, therefore, that the invention be limited only by the scope of the claims which follow and that such claims be interpreted as broadly as is reasonable.

Claims

WHAT IS CLAIMED IS:
1. A compound of the formula
Figure imgf000154_0001
(ID where
?2 '"3
where A, is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, or a direct bond;
A2 is C(0)NR7, NR7C(0), S02NR7, NR7S02, NR7, S, SO, S02, O, or a direct bond, where R7 is as defined below;
A3 is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, or a direct bond;
A4 is alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocyclyl, heteroaryl, aryl, NRgRα, OR8, or hydrogen, where R8 and R9 are as defined below;
Figure imgf000155_0001
where D, is alkylene, alkenylene, alkynylene, NR]0(O)C, NR]0, S, SO, S02, O, or a direct bond, where Rio is as defined below;
D2 is
S, SO, S02, O, C(0)NR„, NR,]C(0), NRπ, or a direct bond, where R,, is as defined below;
D3 is alkylene, alkenylene, alkynylene, arylene, heteroarylene, S, SO, S02, O, C(0)NR]2, NR]2C(0), S02NR,2, NR]2S02, NR,2, or a direct bond, where Rn is as defined below;
D4 is aryl, aryloxy, heteroaryl, or heteroaryloxy;
D5 and D6 are, independently, lower alkylene, O, S, SO, or S02;
R3 is hydrogen or lower alkyl; P is Λ\W>
T2 E^3
I 4
I 6 E7
where Ei is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, C(0)NRι3, or a direct bond, where Rn is as defined below;
E2 is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, NR] , S, SO, S02, O, C(O), or a direct bond, where R,4 is as defined below;
E3 is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, NR,5, S, SO, S02, O, C(O),
Figure imgf000156_0001
or a direct bond, where R]5 and Rι6 are as defined below;
E4 is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, NR,8, S, SO, SO,, O, N(Rl 8)C(0), C(0)N(R,8), C(O),
Figure imgf000156_0002
or a direct bond, where R and R]8 are as defined below;
E5 is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, NR,9, S, SO, S02, O, C(O),
Figure imgf000157_0001
or a direct bond, where Rj9 and R20 are as defined below;
E6 is alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, NR2ι, S, SO, S02, O, C(O), or a direct bond, where R2, is as defined below;
E7 is hydrogen, NR22R23, OR22, SR22, SOR22, S02R22, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocyclyl, aryl, or heteroaryl, where R22 and R23 are as defined below;
R5 is hydrogen or lower alkyl;
Re is
^ Z
Figure imgf000157_0002
where
heteroarylene; Z2 is lower alkylene, lower alkenylene, lower alkynylene, cycloalkylene, cycloalkenylene, arylene, heterocyclylene, heteroarylene, C(0)NR24, NR24C(0), S02NR24, NR24S02, NR24, S, SO, S02, O, C(O), C(0)0, OC(O), or a direct bond, where R24 is as defined below;
Z3 is alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocyclyl, heteroaryl, aryl, NR25R26, OR25, or hydrogen, where R25 and R26 are as defined below; and
R7, R8, R9, Rio, Rn, Ri2> RB, Rι4, Ris, Ri6, i7, Ri8 Ri9, R2o, 21, R22, R23, R24> R25) and R 6 are, independently, hydrogen, alkyl, alkynyl, alkenyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, or heteroaryl;
or a pharmaceutically acceptable salt, solvate, biohydrolyzable ester, biohydrolyzable amide, affinity reagent, or prodrug thereof.
2. A compound of the formula:
Figure imgf000158_0001
CD where
Ri is methyl, trifluoromethyl, ethyl, isopropyl, n-propyl, tert-butyl, 3-methoxycyclopentyl, furan-2-ethynyl, 4-methyl-l -pentyl, 2-thiophenesulfanylmethyl, 4-trifluoromethylcyclohexyl,
3-aminophenoxymethyl, 3-(4-mo holine)-l -propyl, 2-(3-tefrazolyl)-l -ethyl, 2-(3-pyridyl)-l- ethyl, 2-(3-furyl)-l -ethyl, 2-(2-thiazolyl)-l -ethyl, 3,3,3-trιfluoro-l -propyl, 4,4,4-frifluoro- 1 - butyl, 2-(4-tπfluorophenyl)-l -ethyl, thιophene-3 -ethynyl, 2-mtrophenoxymethyl, 3- nitrophenoxymethyl, ethynyl, 2-propynyl, 2-butynyl, phenylethynyl, or vinyl;
R2 ιs 5-methylthιophene-2-methyl, 2-furanmethyl, thιophene-2-methyl, benzothιophene-2-methyl, benzofuran-2-methyl, 4-fluorobenzyl, 3-phenyl-l -propyl, 3-phenyl-2-methyl-l -propyl, 3-(2- pyπdyl)-l -propyl, 3-(thιophene-2-yl)-l-propyl, 4-phenyl-l -butyl, 3-phenyl-2-propene-l-yl, 3-(benzofuran-3-yl)-l -propyl, 3-(benzothιophene-3-yl)-l -propyl, 3-(furan-2-yl)-l -propyl, 3- (2-thιazolyl)-l -propyl, 3 -(pyπmιdm-2-yl)-l -propyl, 3-phenyl-2-ethyl-l -propyl, benzyloxymethyl, 2-benzyloxy- 1 -ethyl, 3-(3-pyπdyl)-l -propyl, 2-phenyl-l -ethyl, 3- benzyloxy-1 -propyl. 4-phenylcyclohexylmethyl, 3-(furan-3-yl)-l -propyl, 1,2,3,4- tefrahydronaphthalene-2-methyl, 4-bιphenylpropyl, 3-phenyl-l -butyl, 2,3- dιhydrobenzo[l,4]dιoxιne-2-methyl, 2-naphthylmethyl, chroman-2-methyl, 3-phenyl-2- methyl-2-propene-l-yl, 3-bιphenyl, 4-phenyl-3-methyl-2-butyl, 4-(3-thιophenyl)-2-butyl, benzothιophene-3-methyl, benzoxazole-2-methyl, 4-(3-furyl)-2-butyl, 3-(4-chlorophenyl)-l- propyl, 3-phenoxyphenyl-l -propyl, or benzyl;
R3 ιs hydrogen, isobutyl, or methyl;
R4 is tert-butyl, 1-propoxy-l -ethyl, 4-(benzyloxycarbonylammo)-l -butyl, 2-(2- (benzyloxycarbonylamιno)-l-ethylsulfanyl)-2-propyl, 4-(2-pyπdylcarbonylamιno)-2-methyl- 2-butyl, 4-ιsobutoxycarbonylammo-l -butyl, 3-pyπdylmethyl, 3-(2-thιophenecarbonylamιno)- 2-methyl-2-propyl, 4-propoxycarbonylammo-2-butyl, 4-(2-naphthylacetylammo)-l -butyl, 1- ethoxycarbonylammo-1 -ethyl, 4-(2-pyπdylcarbonylamιno)-2-methyl-2-butyl, 3- (benzyloxycarbonylammo)- 1 -propyl, 1 -methanesulfanyl- 1 -ethyl, 3-(2-pyπdylcarbonylammo)-
2-mefhyl-2-propyl, 3-(2-thιophenecarbonylamιno)-2-propyl, 3-carbamoylammo-l -propyl, 4- (4-pyπdylcarbonylammo)-2-methyl-2-butyl, 2-ethoxycarbonylamιno-l -propyl, 4- hydroxybenzyl, 4-chlorobenzyl, l-(tetrahydrofuran-3-yloxy)-l -ethyl, 1 -methanesulfenyl- 1 - ethyl, 4-fluorobenzyl, 3-(ιmιno-benzenesulfonylamιno)-methylamιno-l -propyl, 4- propoxycarbonylammo-1 -butyl, 3-(ιmιno-(2,3,6-tπmefhyl-4- methoxybenzenesulfonylammo))-methylammo- 1 -propyl, 2-(2-( 1 ,3 ,4- thιadιazol)ylammosulfonyl)- 1 -ethyl, 3-methylcarbamoylamιno- 1 -propyl, 4- benzyloxycarbonylaminobenzyl, isopropyl, 2-(3-pyπdylcarbonylammo)-l -ethyl, 1,1- dιmethyl-1 -propyl, 2-(2-fhιophene)-l -ethyl, cyclohexyl, 3-phenylcarbamoylammo-l -propyl, 4-cyclopentylacetylammo-l -butyl, 4-(3-methoxybenzoylamιno)-l -butyl, 4- ethoxycarbonylamino- 1 -butyl, 4-ethoxycarbonylammo- 1 -butyl, 2-(2-(ethoxycarbonylammo)- l-ethylsulfanyl)-2-propyl, 2-butyl, 1-methoxy-l -ethyl, 1 -hydroxy- 1 -ethyl, 2-(methoxy methylaminocarbonyl)- 1 -ethyl, 2-(4-ethoxycarbonyl- 1 -piperazmecarbonyl)- 1 -ethyl, 2- guanιdιnesulfonyl-1 -ethyl, 2-methyl-4-(2-pyπdylcarbonylamιno)-2-butyl, 2-(methyl benzylaminocarbonyl)- 1 -ethyl, 2-(4-moφhohnecarbonyl)- 1 -ethyl, 2- pyπdylcarbonylammomethyl, acetylammomethyl, 1 -isobutoxy- 1 -ethyl, carbamoylammomethyl, dimethylammocarbonylmethyl, 2-dιmethylamιnosulfonyl-l -ethyl, 2- methanesulfanyl-2 -propyl, 2-hydroxy-2-propyl, 4-(2-pyπdylcarbonylamιno- 1 -butyl, 2- (dimethylaminocarbonyl)- 1 -ethyl, 2-methanesulfonyl- 1 -ethyl, 1 -(2-pyπdylmethoxy)- 1 -ethyl, 1-benzyloxy-l -ethyl, phenyl, 2-methyl-l -propyl, or 2-phenyl-l -ethyl;
R5 ιs hydrogen, methyl, ethyl, or propyl; and
R& is 2-fhιazolyl, 2-pyπdyl, 2-pyπmιdιnyl, 2-(l,3,4-thιadιazolyl), 3-(5-mefhylιsoxazolyl), 3- pyπdyl, 3-mdolyl, 2-(5-methylthιazolyl), 3-hydroxy-2-pyπdyl, 2-(5-ethanesulfanyl- 1,3,4- thiadiazolyl), 2-benzothιazolyl, 6-methoxy-2-benzothιazolyl, 4-pyπdyl, 2-pyrazmyl, 3- quinohnyl, 2-(5-methyl-l,3,4-thιadιazolyl), 6-amιno-3-pyndyl, 6-amιno-2-pyπdyl, 4-ammo-
3-pyπdyl, or 3-amιno-2-pyπdyl; or a pharmaceutically acceptable salt, solvate, biohydrolyzable ester, biohydrolyzable amide, affinity reagent, or prodrug thereof.
A compound of the formula:
Figure imgf000161_0001
(II) where Ri , R3, R4, R5 and R are as defined above in claim 2 and where R2 is 5-phenyl-l-
pentyl, 3-(4-tπfluorophenyl)-l-propyl, 3-(4-methylphenyl)-l -propyl, 3-(4-phenoxyphenyl)-l- propyl, 3 -(4-tert-butylphenyl)-l -propyl, 3-(4-methoxyphenyl)-l -propyl, 3-(4-tπfluorophenyl)- 1 -propyl, or 3-(3-phenoxyphenyl)-l -propyl; or a pharmaceutically acceptable salt, solvate, biohydrolyzable ester, biohydrolyzable amide, affinity reagent, or prodrug thereof.
4. A compound of the formula:
Figure imgf000161_0002
(II) where R, is methyl, ethyl, n-propyl, 4-methyl-l -pentyl, 2-thiophenesulfanylmethyl, 3- ammophenoxymethyl, 2-(3 -tetrazolyl)- 1 -ethyl, 2-(3-pyπdyl)-l -ethyl, 2-(3-furyl)-l -ethyl, 2- (2-thιazolyl)-l -ethyl, 3,3,3-tπfluoro-l-propyl, 2-(4-tπfluorophenyl)-l -ethyl, thιophene-3- ethynyl, 2-mtrophenoxymethyl, 3-nιtrophenoxymethyl, or vinyl;
R2 ιs
5-methylthιophene-2-methyl, 2-furanmethyl, thιophene-2-methyl, benzothιophene-2-methyl, benzofuran-2-methyl, 4-fluorobenzyl, 3-phenyl-l-propyl, 3-phenyl-2-methyl-l -propyl, 3-(2- pyndyl)-l -propyl, 3-(thιophene-2-yl)-l -propyl, 4-phenyl-l -butyl, 3-phenyl-2-propene-l-yl, 3-(benzofuran-3-yl)-l -propyl, 3-(benzothιophene-3-yl)-l -propyl, 3-(furan-2-yl)-l -propyl, 3- (2-thιazolyl)- 1 -propyl, 3-(pyπmιdιn-2-yl)- 1 -propyl, 3-phenyl-2-ethyl- 1 -propyl, benzyloxymethyl, 2-benzyloxy- 1 -ethyl, 3-(3-pyπdyl)-l -propyl, 2-phenyl-l -ethyl, 3- benzyloxy-1 -propyl. 4-phenylcyclohexylmethyl, 3-(furan-3-yl)-l-propyl, 1,2,3,4- tetrahydronaphthalene-2-methyl, 3-phenyl- 1 -butyl, 2,3-dιhydrobenzo[ 1 ,4]dιoxιne-2-methyl, 2-naphthylmethyl, chroman-2-methyl, 3-phenyl-2-methyl-2-propene-l-yl, 4-phenyl-3- methyl-2-butyl, 4-(3-thιophenyl)-2-butyl, benzothιophene-3-methyl, benzoxazole-2-methyl, 4-(3-furyl)-2-butyl, 3-(4-chlorophenyl)-l -propyl, or benzyl;
R3 ιs hydrogen;
R» is tert-butyl, 4-(benzyloxycarbonylammo)- 1 -butyl, 2-(2-(benzyloxycarbonylamιno)- 1 - ethylsulfanyl)-2 -propyl, 3-pyπdylmethyl, 4-(2-naphthylacetylamιno)- 1 -butyl, 3- (benzyloxycarbonylamino)- 1 -propyl, 3-carbamoylammo-l -propyl, 4-hydroxybenzyl, 4- chlorobenzyl, 4-fluorobenzyl, 3-(ιmιno-(2,3,6-frιmethyl-4-methoxybenzenesulfonylammo))- methylammo-1 -propyl, 4-benzyloxycarbonylamιnobenzyl, isopropyl, cyclohexyl, 4- cyclopentylacetylammo- 1 -butyl, 4-(3-methoxybenzoylamιno)- 1 -butyl, 4- efhoxycarbonylamino- 1 -butyl, 2-(2-(efhoxycarbonylammo)- 1 -ethylsulfanyl)-2-propyl, 2- butyl, 1-methoxy-l -ethyl, 1 -hydroxy- 1 -ethyl, isopropyl, 2-(methoxy methylaminocarbonyl)- 1 -ethyl, 2-(4-ethoxycarbonyl-l-pιperazιnecarbonyl)-l -ethyl, 2-guanιdmesulfonyl-l -ethyl, 2- methyl-4-(2-pyπdylcarbonylamιno)-2-butyl, 2-(methyl benzylammocarbonyl)- 1 -ethyl, 2-(4- moφholmecarbonyl)- 1 -ethyl, 2-pyπdylcarbonylamιnomethyl, acetylaminomethyl, 1- lsobutoxy- 1 -ethyl, carbamoylammomethyl, dimethylammocarbonylmethyl, 2- dimethylaminosulfonyl- 1 -ethyl, 2-methanesulfanyl-2-propyl, 2-hydroxy-2-propyl, 4-(2- pyπdylcarbonylamino- 1 -butyl, 2-(dιmethylammocarbonyl)- 1 -ethyl, 2-methanesulfonyl- 1 - ethyl, l-(2-pyπdylmethoxy)-l -ethyl, 1-benzyloxy-l -ethyl, phenyl, 2-methyl-l -propyl, or 2- phenyl- 1 -ethyl;
R5 ιs hydrogen; and
Re is
2-thιazolyl, 2-pyπdyl, 2-pyrιmιdmyl, 2-(l,3,4-thιadιazolyl), 3-(5-methylιsoxazolyl), 3- pyπdyl, 2-(5-methylthιazolyl), 3-hydroxy-2-pyπdyl, 2-(5-ethanesulfanyl-l,3,4-thιadιazolyl), 4-pyπdyl, 2-pyrazιnyl, 2-(5-methyl-l,3,4-thιadιazolyl), 6-ammo-3-pyπdyl, 6-amιno-2- pyπdyl, 4-ammo-3-pyπdyl, or 3-ammo-2-pyπdyl;
or a pharmaceutically acceptable salt, solvate, biohydrolyzable ester, biohydrolyzable amide, affinity reagent, or prodrug thereof.
5. A compound of the formula:
Figure imgf000164_0001
(II) where Ri , R3, R4, R5 and Rg are as defined above in claim 4 and where R2 is 5-phenyl-l-
pentyl, 3-(4-trifluorophenyl)-l -propyl, 3 -(4-methylphenyl)-l -propyl or 3-(4-phenoxyphenyl)- 1 -propyl; or a pharmaceutically acceptable salt, solvate, biohydrolyzable ester, biohydrolyzable amide, affinity reagent, or prodrug thereof.
6. A compound of the formula:
Figure imgf000164_0002
(I") where
R, is methyl, n-propyl, isopropyl, ethyl, or 3,3,3-trifluoro-l-propyl;
R2 is
5 -methyl thiophene-2-methyl, 3-phenyl-l -propyl, 4-phenyl-l -butyl, 2-phenyl-l -ethyl,. 4- phenylcyclohexylmethyl, 3-(4-chlorophenyl)-l -propyl, or benzyl;
R3 is hydrogen; P IS tert-butyl, isopropyl, 2-butyl, 1-methoxy-l -ethyl, 1 -hydroxy- 1 -ethyl, 2-mefhanesulfanyl-2- propyl, or 2-hydroxy-2-propyl;
R5 is hydrogen; and
Re is 2-thiazolyl, 2-pyridyl, 3-pyridyl, 4-pyridyl or 2-(l,3,4-thiadiazolyl); or a pharmaceutically acceptable salt, solvate, biohydrolyzable ester, biohydrolyzable amide, affinity reagent, or prodrug thereof.
7. A compound of the formula:
Figure imgf000165_0001
(ID where Ri , R3, R4, R5 and Rg are as defined above in claim 6 and where R2 is 5-phenyl-l-
pentyl, 3-(4-trifluorophenyl)-l -propyl, 3-(4-methylphenyl)-l -propyl or 3-(4-phenoxyphenyl)- 1 -propyl; or a pharmaceutically acceptable salt, solvate, biohydrolyzable ester, biohydrolyzable amide, affinity reagent, or prodrug thereof.
8. A compound of claim 1, wherein the compound is selected from the examples listed in
Table 2A.
9. A compound of claim 1, wherein the compound is selected from the examples listed in
Table 2B.
10. A compound of claim 1, wherein the compound is selected from the examples listed in Table 3.
11. A compound of claim 1 , wherein the compound is selected from the examples listed in Table 4.
12. A compound of formula (II) as claimed in any one of claims 1 to 11 for use in therapy.
13. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a pharmacologically effective amount of a compound as claimed in any one of claims 1 to 11.
14. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a pharmacologically effective amount of the compound as claimed in any one of claims 1 to 1 1 sufficient to inhibit the cellular release of mature tumor necrosis factor alpha.
15. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a pharmacologically effective amount of the compound as claimed in any one of claims 1 to 11 sufficient to inhibit a matrix metalloprotease.
16. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a pharmacologically effective amount of the compound as claimed in any one of claims 1 to
1 1 sufficient to inhibit the shedding of cell surface protein ectodomains.
17. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a pharmacologically effective amount of the compound as claimed in any one of claims 1 to 11 sufficient to inhibit CD23 proteolysis.
18. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a pharmacologically effective amount of the compound as claimed in any one of claims 1 to 11 sufficient to inhibit the growth of tumor metastases.
19. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a pharmacologically effective amount of the compound as claimed in any one of claims 1 to 11 sufficient to treat diabetes.
20. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a pharmacologically effective amount of the compound as claimed in any one of claims 1 to 11 sufficient to treat arthritis.
21. A method of inhibiting the intracellular release of tumor necrosis factor alpha, comprising the step of administering to a mammal in need thereof a pharmacologically effective amount of a compound as claimed in any one of claims 1 to l l.
22. A method of inhibiting a matrix metalloprotease, comprising the step of administering to a mammal in need thereof a pharmacologically effective amount of a compound as claimed in any one of claims 1 to 11.
23. A method of inhibition of shedding of cell surface protein ectodomains, comprising the step of administering to a mammal in need thereof a pharmacologically effective amount of a compound as claimed in any one of claims 1 to 11.
24. A method of inhibition of CD23 proteolysis, comprising the step of administering to a mammal in need thereof a pharmacologically effective amount of a compound as claimed in any one of claims 1 to 11.
25. The use of a compound as claimed in any one of claims 1 to 11 in the preparation of a medicament to inhibit the cellular release of mature tumor necrosis factor alpha.
26. The use of a compound as claimed in any one of claims 1 to 11 in the preparation of a medicament to inhibit a matrix metalloprotease.
27. The use of a compound as claimed in any one of claims 1 to 11 in the preparation of a medicament to inhibit the shedding of cell surface protein ectodommains.
28. The use of a compound as claimed in any one of claims 1 to 11 in the preparation of a medicament to inhibit CD23 proteolysis.
29. The use of a compound as claimed in any one of claims 1 to 11 in the preparation of a medicament to inhibit the growth of tumor metastases.
30. The use of a compound as claimed in any one of claims 1 to 11 in the preparation of a medicament to treat diabetes.
1. The use of a compound as claimed in any one of claims 1 to 11 in the preparation of a medicament to freat arthritis.
PCT/US1999/018960 1998-08-26 1999-08-25 Formamide compounds as therapeutic agents WO2000012082A1 (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1121118A1 (en) * 1998-08-26 2001-08-08 Glaxo Group Limited Formamides as therapeutic agents

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1997003783A1 (en) * 1995-07-20 1997-02-06 British Biotech Pharmaceuticals Limited Metalloproteinase inhibitors

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1997003783A1 (en) * 1995-07-20 1997-02-06 British Biotech Pharmaceuticals Limited Metalloproteinase inhibitors

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1121118A1 (en) * 1998-08-26 2001-08-08 Glaxo Group Limited Formamides as therapeutic agents
EP1121118A4 (en) * 1998-08-26 2002-09-11 Glaxo Group Ltd Formamides as therapeutic agents

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