WO 99/52910
`
`PCT/1IB99/00503
`
`-9.
`
`leukemias and lymphomas), tissue ulceration, restenosis, periodontal disease, epidermolysis
`
`bullosa,
`
`osteoporosis,
`
`loosening of
`
`artificial
`
`joint
`
`implants,
`
`atherosclerosis
`
`(including
`
`atherosclerotic plaque rupture), aortic aneurysm (including abdominal aortic aneurysm and brain
`
`aortic aneurysm), congestive heart failure, myocardial infarction, stroke, cerebral ischemia, head
`trauma, spinal cord injury, neuro-degenerative disorders (acute and chronic), autoimmune
`disorders, Huntington’s disease, Parkinson’s disease, migraine, depression, periphéral
`
`10
`
`neuropathy,
`
`pain,
`
`cerebral
`
`amyloid
`
`angiopathy,
`
`nootropic or
`
`cognition
`
`enhancement,
`
`amyotrophic fateral sclerosis, muitiple sclerosis, ocular angiogenesis, corneal injury, macular
`
`degeneration, abnormal wound healing, burns, diabetes, tumor invasion, tumor growth, tumor
`
`metastasis, corneal
`
`scarring,
`
`scleritis, AIDS,
`
`sepsis,
`
`septic shock and other diseases
`
`15
`
`characterized by metalloproteinase activity and other diseases characterized by mammalian
`
`reprolysin activity in a mammal,
`
`including a human, comprising an amount of a compound of
`
`formula | or a pharmaceutically acceptable salt thereof effective in such treatments and a
`
`pharmaceutically acceptable carrier.
`
`The present invention also relates to a pharmaceutical composition for the inhibition of
`
`20
`
`(a) matrix metalloproteinases or other metalloproteinases involved in matrix degradation, or (b) a
`
`mammalian reprolysin (such as aggrecanase or ADAM’s TS-1, 10, 12, 15 and 17, most
`
`preferably ADAM-17) in a mammal,
`
`including a human, comprising an effective amount of a
`
`compoundof formula I or a pharmaceutically acceptable sait thereof.
`
`The presentinvention also relates to a method for treating a condition selected from the
`
`25
`
`group consisting of arthritis (including osteoarthritis and rheumatoid arthritis), inflammatory bowel
`
`disease, Crohn's disease, emphysema, chronic obstructive pulmonary disease, Alzheimer’s
`
`disease, organ transplant toxicity, cachexia, allergic reactions, allergic contact hypersensitivity,
`
`cancer, tissue ulceration, restenosis, periodontal disease, epidermolysis bullosa, osteoporosis,
`
`loosening of artificial joint implants, atherosclerosis (including atherosclerotic plaque rupture),
`
`30
`
`aortic aneurysm (including abdominai aortic aneurysm and brain aortic aneurysm), congestive
`
`heart failure, myocardial infarction, stroke, cerebral ischemia, head trauma, spinal cord injury,
`
`neuro-degenerative disorders (acute and chronic), autoimmune disorders, Huntington's disease,
`
`Parkinson's disease, migraine, depression, peripheral neuropathy, pain, cerebral amyloid
`angiopathy, nootropic or cognition enhancement, amyotrophic lateral
`sclerosis, multiple
`sclerosis, ocular angiogenesis, corneal injury, macular degeneration, abnormal wound healing,
`
`35
`
`burns, diabetes,
`
`tumor invasion,
`
`tumor growth,
`
`tumor metastasis, comeal scarring, scleritis,
`
`AIDS, sepsis, septic shock and other diseases characterized by metalloproteinase activity and
`
`other diseases characterized by mammalian reprolysin activity in a mammal, including a human,
`
`comprising administering to said mammal an amount of a compound of formula | or a
`
`40
`
`pharmaceutically acceptable salt thereof effective in treating such a condition.
`
`AQUESTIVE EXHIBIT 1007
`
`AQUESTIVE EXHIBIT 1007 page 2001
`
`page 2001
`
`

`

`WO 99/52910
`
`PCT/1B99/00503
`
`-10-
`
`The present
`
`invention also relates to a method for
`
`the inhibition of
`
`(a) matrix
`
`metalloproteinases or other metalloproteinases involved in matrix degradation, or
`
`(b) a
`
`mammalian reprolysin (such as aggrecanase or ADAM's TS-1, 10, 12, 15 and 17, preferably
`
`including a human, comprising administering to said mammal an
`ADAM-17) in a mammal,
`effective amount of a compoundof formula | or a pharmaceutically acceptable salt thereof.
`This invention also encompasses pharmaceutical compositions containing prodrugsof
`compoundsofthe formula |. This invention also encompasses methodsoftreating or preventing
`disorders that can be treated or prevented by the inhibition of matrix metalloproteinases or the
`inhibition of mammalian reprolysin comprising administering prodrugs of compounds of the
`formula |. Compoundsof formula | having free amino, amido, hydroxy or carboxylic groups can
`be converted into prodrugs. Prodrugs include compounds wherein an amino acid residue, or a
`
`polypeptide chain of two or more (e.g.,
`
`two, three or four) amino acid residues which are
`
`covalently joined through peptide bonds to free amino, hydroxy or carboxylic acid groups of
`compoundsofformula |. The amino acid residues include the 20 naturally occurring amino acids
`commonly designated by three letter symbols and also include, 4-hydroxyproline, hydroxylysine,
`demosine,
`isodemosine, 3-methylhistidine, norvalin, beta-alanine, gamma-aminobutyric acid,
`
`citrulline, homocysteine, homoserine, ornithine and methionine sulfone. Prodrugs aiso include
`
`compounds wherein carbonates, carbamates, amides and alkyl esters which are covalently
`
`bondedto the above substituents of formula ! through the carbony! carbon prodrug sidechain.
`
`One of ordinary skill in the art will appreciate that the compoundsof the invention are
`
`10
`
`15
`
`20
`
`25
`
`useful
`
`in treating a diverse array of diseases. One of ordinary skill
`
`in the art will also
`
`appreciate that when using the compounds of the invention in the treatment of a specific
`
`disease that
`
`the compounds of the invention may be combined with various existing
`
`therapeutic agents used for that disease.
`
`For the treatment of rheumatoid arthritis, the compounds of the invention may be
`
`30
`
`combined with agents such as TNF-a inhibitors such as anti-TNF monoclonal antibodies and
`
`TNF receptor
`
`immunoglobulin molecules (such as Enbrel®),
`
`low dose methotrexate,
`
`lefunimide, hydroxychloroquine, d-penicilamine, auranofin or parenteral or oral gold.
`
`The compounds of the invention can also be used in combination with existing
`
`therapeutic agents for
`
`the treatment of osteoarthritis.
`
`Suitable agents to be used in
`
`35
`
`combination include standard non-steroidal anti-inflammatory agents (hereinafter NSAID's)
`
`such as piroxicam, diclofenac, propionic acids such as naproxen, flubiprofen, fenoprofen,
`
`ketoprofen and ibuprofen, fenamates such as mefenamic acid,
`
`indomethacin, sulindac,
`
`apazone, pyrazolones such as phenylbutazone, salicylates such as aspirin, COX-2 inhibitors
`
`such as
`
`celecoxib and rofecoxib,
`
`analgesics and intraarticular
`
`therapies
`
`such as
`
`40
`
`corticosteroids and hyaluronic acids such as hyalgan and synvisc.
`
`AQUESTIVE EXHIBIT 1007
`
`AQUESTIVE EXHIBIT 1007 page 2002
`
`page 2002
`
`

`

`WO 99/52910
`
`PCT/1B99/00503
`
`-1l1-
`
`The compounds of the present invention may also be used in combination with
`
`anticancer agents such as endostatin and angiostatin or cytotoxic drugs such as adriamycin,
`
`daunomycin, cis-platinum, etoposide, taxoi, taxotere and alkaloids, such as vincristine, and
`
`antimetabolites such as methotrexate.
`
`The compounds of the present invention may also be used in combination with
`cardiovascular agents such as calcium channel blockers,
`lipid lowering agents such as
`statins, fibrates, beta-blockers, Ace inhibitors, Angiotensin-2 receptor antagonists and platelet
`
`aggregation inhibitors.
`
`The compounds of the present invention may also be used in combination with CNS
`
`agents such as antidepressants (such as sertraline), anti-Parkinsonian drugs (such as
`deprenyl, L-dopa, requip, miratex, MAOBinhibitors such as selegine and rasagiline, comP
`inhibitors such as Tasmar, A-2 inhibitors, dopamine reuptake inhibitors, NMDA antagonists,
`
`Nicotine agonists, Dopamine agonists and inhibitors of neuronal nitric oxide synthase), and
`anti-Alzheimer's drugs
`such as Aricept,
`tacrine, COX-2 inhibitors, propentofylline or
`
`metryfonate.
`
`The compounds of the present invention may also be used in combination
`
`with osteoporosis agents such as droloxifene or fosomax and immunosuppressant agents
`
`such as FK-506 and rapamycin.
`
`Detailed Description of the Invention
`
`10
`
`15
`
`20
`
`The following reaction Schemesillustrate the preparation of the compounds of the
`present invention. Unless otherwise indicated n, R', R*, Q and Z in the reaction Schemes and
`
`25
`
`the discussion that follow are defined as above.
`
`AQUESTIVE EXHIBIT 1007
`
`AQUESTIVE EXHIBIT 1007 page 2003
`
`page 2003
`
`

`

`WO 99/52910
`
`PCT/1IB99/00503
`
`-12-
`
`SCHEME1
`
`O
`
`LooYo
`
`IX
`
`O
`
`HO
`
`OH
`
`Vi
`
`°
`
`oO
`
`V
`
`Oo
`
`Oo
`
`1
`
`PG O
`
`1
`
`OPG
`
`Vii
`
`oO~e1"
`
`VII
`
`QO
`
`HO
`
`S—Q
`
`IV
`
`AQUESTIVE EXHIBIT 1007
`
`AQUESTIVE EXHIBIT 1007 page 2004
`
`page 2004
`
`

`

`WO 99/52910
`
`PCT/1B99/00503
`
`-13-
`
`SCHEME 1 CONTINUED
`
`S-Q
`
`~
`
`o
`$-Q
`
`O
`
`Oo
`Zz.||
`S—aQ
`
`|O
`
`C1 i
`
`Or
`
`N
`H
`
`HHI
`
`CL l
`
`Oo.
`
`N
`
`i
`
`O
`
`HO.
`
`N
`H
`
`AQUESTIVE EXHIBIT 1007
`
`AQUESTIVE EXHIBIT 1007 page 2005
`
`page 2005
`
`

`

`WO 99/52910
`
`PCT/1B99/00503
`
`-14-
`
`SCHEME2
`
`O
`
`PG’O
`
`cS
`
`XIV
`
`O
`
`PG’0
`
`NH,
`
`Xill
`
`foalH
`>i
`
`P60
`
`Xil
`
`AQUESTIVE EXHIBIT 1007
`
`AQUESTIVE EXHIBIT 1007 page 2006
`
`page 2006
`
`

`

`WO 99/52910
`
`PCT/IB99/00503
`
`-15-
`
`SCHEME 2 CONTINUED
`
`|
`
`O
`
`HO
`
`zit
`
`xl
`
`Cl
`
`oO.
`
`HO
`
`O
`ao
`N-S-Q
`
`N
`
`Xx
`
`I
`N-$
`res
`:
`
`x
`
`1
`on
`
`AQUESTIVE EXHIBIT 1007
`
`AQUESTIVE EXHIBIT 1007 page 2007
`
`page 2007
`
`

`

`WO 99/52910
`
`PCT/1IB99/00503
`
`-16-
`
`SCHEME3
`
`9
`
`PG’0
`
`N—so,a
`
`Xl
`
`5
`
`N—SO,Q
`
`XVII
`
`Oo
`
`R'
`N—SO,Q
`
`HO
`
`XVI
`
`O
`
`R'
`
`or
`
`N—SO,Q
`
`XV
`
`>
`
`|
`
`oy
`
`Z—SO,Q
`
`AQUESTIVE EXHIBIT 1007
`
`AQUESTIVE EXHIBIT 1007 page 2008
`
`page 2008
`
`

`

`WO 99/52910
`
`PCT/IB99/00503
`
`SCHEME 4
`
`Oo
`
`PG2o
`
`N—so,a
`
`Xll
`
`XIX
`
`XVIII
`
`CO,R?
`
`a N
`
`—SO,Q
`
`°
`
`|
`
`O
`
`cr°
`
`CO,R”
`
`O
`
`HO
`
`N—SO,Q
`
`
`
`oO
`
`ON
`
`Z—SO,Q
`
`AQUESTIVE EXHIBIT 1007
`
`AQUESTIVE EXHIBIT 1007 page 2009
`
`page 2009
`
`

`

`WO 99/52910
`
`PCT/IB99/00503
`
`-18-
`
`Scheme 1 refers to the preparation of compounds of formula |, wherein Z is CH,.
`
`Referring to Scheme |, a compound of the formula |
`
`is prepared from a compound of the
`
`formula Il by hydrogenolysis under an atmosphere of hydrogen in the presence of a catalyst in
`
`a reaction inert solvent. Suitable catalysts include 5% palladium on barium sulfate or 5%
`
`palladium on carbon, preferably 5% palladium on barium sulfate. Suitable solvents include an
`
`10
`
`alcohol such as ethanol, methanol or isopropanol, preferably methanol. The aforéSaid
`
`reaction may be performed at a pressure from about 1
`
`to about 5 atmospheres, preferably
`
`about 3 atmospheres. Suitable temperatures for the aforesaid reaction range from about
`
`20°C (room temperature) to about 60°C, preferably the temperature may range from about
`
`20°C to about 25°C (i.e. room temperature). The reaction is complete within about 0.5 hours
`
`15
`
`to about 5 hours, preferably about 3 hours.
`
`Compoundsof the formula Il can be prepared from compoundsof the formula HI by
`
`reaction with an oxidant
`
`in a reaction inert solvent.
`
`Suitable oxidants include meta-
`
`chloroperbenzoic
`
`acid,
`
`hydrogen
`
`peroxide
`
`or
`
`sodium perborate,
`
`preferably meta-
`
`chloroperbenzoic acid. Suitable solvents include halogenated solvents such as methylene
`
`20
`
`chloride or chloroform, preferably methyiene chloride. Suitable temperatures for the aforesaid
`
`reaction range from about 0°C to about 60°C, preferably the temperature may range from
`
`about 20°C to about 25°C (i.e. room temperature). The reaction is complete within about 0.5
`
`hours to about 24 hours, preferably about 16 hours.
`
`The compound of formula Ill is prepared from a compound of formula IV by reaction
`
`25
`
`with O-benzylhydroxyamine hydrochloride, an activating agent, and a base in a reaction inert
`
`solvent.
`
`Suitable
`
`activating
`
`agents
`
`include
`
`(benzotriazol-1-yloxy)tris(dimethylamino)
`
`phosphonium hexafluorophosphate
`or
` 1-(3-(dimethylaminopropyl)-3-ethylcarbodiimide
`hydrochloride,
`preferably
`(benzotriazol-1-yloxy)tris(dimethylamino)
`phosphonium
`
`hexafluorophosphate. such as_triethylamine,Suitable bases include tertiary amines
`
`30
`
`diisopropylethylamine or 4-N,N-dimethylaminopyridine, preferably diisopropylethylamine. The
`
`temperature of the aforesaid reaction may range from about 0°C to about 60°C, preferably
`
`about 50°C. Suitable solvents include N,N-dimethyiformamide, halogenated solvents such as
`
`methylene chloride or chloroform, or ethers such as THF or diethyl ether; preferably the
`
`solvent is N,N-dimethylformamide. The reaction is complete in about 4 hours to about 48
`
`35
`
`hours, preferably about 16 hours.
`
`Compounds of the formula IV, can be prepared from compounds of the formula V, by
`
`reaction with a compound of the formula QSH, wherein Q is as defined above, in the presence
`
`of a strong base in an aprotic polar solvent. Suitable bases include sodium hydride,lithium
`
`diisopropylamide, potassium t-butoxide, sodium amide or potassium hydride, preferably
`
`40
`
`sodium hydride.
`
`Suitable solvents include ethers (such as THF, diethyl ether or 1,2-
`
`AQUESTIVE EXHIBIT 1007
`
`AQUESTIVE EXHIBIT 1007 page 2010
`
`page 2010
`
`

`

`WO 99/52910
`
`PCT/1IB99/00503
`
`-19-
`
`dimethoxyethane), or N, N-dimethyiformamide, preferably the solvent is THF. The aforesaid
`
`reaction is conducted at about -78°C to about 0°C, preferably at about 22°C (i.e.,
`
`room
`
`temperature) for a period of 30 minutes to about 24 hours, preferably about 2 hours.
`
`Compounds of the formula V are prepared from compounds of the formula VI by
`
`dehydration in the presence of a tertiary amine base, preferably triethylamine, optionally in the
`presence of 4-dimethylaminopyridine, and a dehydrating agent in an inert solvent. Suitable
`
`10
`
`dehydrating agents include trifluoromethanesulfonic anhydride, methanesulfonic anhydride,
`
`methanesulfonyl chloride, p-toluenesulfonyl chioride or benzenesulfonyl chloride, preferably
`
`benzenesulfony! chloride. Suitable solvents include diethyl ether or dichloromethane. The
`
`reaction is performed at a temperature from about -80°C to about O°C, preferably about O°C.
`
`15
`
`The reaction is carried out for about 10 minutes to 4 hours, preferably about 1 hour.
`
`The compounds of the formula VI are prepared from a compound of formula VII,
`wherein PG' is methyl! or ethyl, by saponification with a base, such aslithium hydroxide, in a
`solvent mixture. Suitable soivent mixtures include water and methanol or water, methanol
`
`and THF. The reaction is performed at a temperature from about 60°C to about 120°C,
`
`20
`
`preferably at about the reflux temperature of the solvent mixture used. The reaction is carried
`
`out for about 30 minutes to 24 hours, preferably about 16 hours.
`
`The exo-hydroxymethy! isomer of the compound of the formula Vil
`
`is prepared from a
`
`compoundof formula VIII.
`
`In general, a solution of a compound of formula VIII is dissolved in
`
`an inert aromatic solvent, preferably benzeneor toluene, and cooled at about -40° C to -20°C,
`
`25
`
`preferably about -40°C. To the coid solution is added a suitable hindered reducing agent,
`
`preferably disobutylatuminum hydride,
`
`in an inert aromatic
`
`solvent, maintaining the
`
`temperature below -25°C. After the addition is complete, the reaction is maintained below 0°C
`
`for about 3 hours. At about -15°C, a protic solvent, preferably ethanol, is added. Afterstirring
`
`at about -15°C for about 1 hour, sodium borohydride is added and the reaction is allowed to
`
`30
`
`warm to about room temperature while stirring for a period of 2 to 24 hours, preferably about
`16 hours.
`
`The endo-hydroxymethyl isomer of the compound of the formula VII can be prepared
`
`from the exo-hydroxymethyl compound of the formula VI by a series of steps which can invert
`
`the sterochemistry about the carbon atom bearing the hydroxymethyl and carboxylic acid
`
`35
`
`groups.
`
`Specifically,
`
`the exo-hydroxymethyi
`
`isomer of formula VI
`
`is first converted to the
`
`corresponding benzyl! ester. Subsequent Jones oxidation of the alcohol to the carboxylic acid
`
`and alky! ester formation (methyl or ethyl) provides an intermediate mixed benzyl alkyl ester(i.e.
`
`the exo ester is methyl or ethyl and the endo ester is benzy!). The benzy! ester is then removed
`
`by hydrogenolysis and the resulting carboxylic acid is reduced to the alcohol by diborane
`
`40
`
`reduction, providing the endo-hydroxymethyl isomer of the compoundof the formula Vil.
`
`AQUESTIVE EXHIBIT 1007
`
`AQUESTIVE EXHIBIT 1007 page 2011
`
`page 2011
`
`

`

`WO 99/52910
`
`PCT/1B99/00503
`
`-20-
`
`10
`
`15
`
`20
`
`25
`
`is ethyl or methyl, are prepared from
`The compounds formula Vill, wherein PG'
`compounds of the formula IX, wherein L is methanesulfony!, benzenesulfonyl or tosyl, by
`reaction with dimethyl! or diethyl malonate in the presence of a strong base, such as sodium
`hydride, in a polar solvent, such as N,N-dimethyiformamide, for a time period between about 4
`hours to about 24 hours, preferably about 16 hours. The aforesaid reaction temperature is
`between about 70°C to about 150°C, preferably about 140° C.
`~
`Compoundsof the formula IX are known or can be made by methods well knownto
`thoseof ordinary skill in the art.
`
`Compoundsof the formula QSH can be prepared by reaction of an alkyl or ary! halide
`with sodium sulfhydride as described in Jerry March, Advanced Organic Chemistry , 360 and 589
`(3rd ed., 1985). Alternatively, compounds of the formula QSH canalso be prepared by reaction
`of an ary! diazonium salt with sodium sulfhydride as described in March id. at 601. Alternatively,
`compounds ofthe formula QSH can also be prepared by reaction of a Grignard reagent with
`sulfur as described in Marchid. at 550. Alternatively, compoundsof the formula QSH canalso
`be prepared by reduction of a sulfonyi chloride, sulfonic acid or disulfide as described in March
`
`id. at 1107 and 1110.
`Scheme2 refers to the preparation of compoundsof the formula |, wherein Z is >NR‘,
`is hydrogen. Referring to Scheme 2, compounds of formula | can be prepared from
`and R'
`compounds of the formula X by hydrogenolysis under an atmosphere of hydrogen in the
`
`presenceof a catalyst in a reaction inert solvent. Suitable catalysts include 5% palladium on
`barium sulfate or 5% palladium on carbon, preferably 5% palladium on barium sulfate.
`Suitable solvents include an alcohol such as ethanol, methanol or isopropanol, preferably
`methanol. The aforesaid reaction may be performed at a pressure from about 1
`to about 5
`
`atmospheres, preferably about 3 atmospheres.
`
`Suitable temperatures for the aforesaid
`
`reaction range from about 20°C (room temperature) to about 60°C, preferably the temperature
`
`may range from about 20°C to about 25°C (i.e. room temperature). The reaction is complete
`within about 0.5 hours to about 5 hours, preferably about 3 hours.
`
`The compound of formula X is prepared from a compound of the formula XI by
`reaction with O-benzylhydroxylamine hydrochioride in the presence of a catalyst and a base
`
`
`
`
`in inert—solvent.a reaction Suitable catalysts include (benzotriazol-1-
`
`
`
`yloxy)tris(dimethylamino)phosphonium hexafluorophosphate or 1-(3-(dimethylaminopropyl)-3-
`ethylcarbodiimide
`hydrochloride,
`preferably
`(benzotriazol-1-yloxy)tris(dimethylamino)
`phosphonium hexafluorophosphate.
`Suitable bases include tertiary amines such as
`triethylamine,
`diisopropylethylamine
`or
`4-N,N-dimethylaminopyridine,
`preferably
`diisopropylethylamine. The aforesaid reaction temperature is from about 0° C to about 60°C,
`preferably about 50° C. Suitable solvents include N,N-dimethylformamide or halogenated
`
`40
`
`AQUESTIVE EXHIBIT 1007
`
`AQUESTIVE EXHIBIT 1007 page 2012
`
`page 2012
`
`

`

`WO 99/52910
`
`PCT/IB99/00503
`
`-21-
`
`solvents
`
`such as methylene chloride or chioroform; preferably,
`
`the solvent
`
`is N,N-
`
`dimethylformamide. The reaction is conducted over a period of about 4 hours to about 48
`
`hours, preferably about 16 hours.
`
`Compounds of the formula X! are prepared from compounds of the formula Xil,
`wherein PG? is methyl or ethyl, by saponification with a base such as sodium hydroxidein a
`solvent mixture such as water and ethanol. The reaction is performed at a temperaturefrom
`about 60°C to about 100°C, preferably at about the reflux temperature of the solvent mixture
`
`used. The reaction is carried out for about 1 day to 10 days, preferably about 6 days.
`The compoundsofthe formula XII, wherein PG?is methyl or ethyl, are prepared from
`compoundsof the formula XIII, wherein PG? is methyl!or ethyl, by reaction with a compound of
`the formula QSO,CI in the presence of a base, such as triethylamine, and a polar solvent.
`Suitable solvents include N,N-dimethylformamide,
`tetrahydrofuran, 1,2-dimethoxyethane,
`dioxane, water or acetonitrile, preferably N,N-dimethylformamide. The reaction mixture is
`stirred at room temperature for a time period between about
`1 hour to about 24 hours,
`
`preferably about 16 hours.
`Compoundsof the formula XI, wherein PG? is methyl or ethyl, are prepared from
`compoundsofthe formula XIV, wherein PG?is methyl or ethyl, by hydrolysis in the presence of
`aqueous mineral! acid and a solvent such as diethyl ether.
`Suitable mineral acids include
`
`hydrochioric and sulfuric acid, preferably hydrochloric acid. The reaction is carried out at a
`
`10
`
`15
`
`20
`
`temperature ranging from about 0°C to 50°C; preferably the temperature may range from about
`20°C to about 25°C (i.e. room temperature). The reaction is conducted over a period of about
`
`25
`
`2 hours to about 48 hours, preferably about 16 hours.
`Compoundsof the formula XIV, wherein PG?is methyl, ethy! or benzyl, are prepared
`from compoundsof the formula IX, wherein L is methanesulfonyl, benzenesulfony! or tosyl, by
`reaction with N-diphenylmethylene glycine, methyl, ethy! or benzyl ester, in the presence of a
`
`30
`
`strong base, such as sodium hydride, in a polar solvent, such as N,N-dimethylformamide, for a
`
`time period between about 4 hours to about 24 hours, preferably about 16 hours. The aforesaid
`
`reaction temperature is between about 70°C to about 140°C, preferably about 100° C.
`Compoundsof the formula XIV, wherein PG? is methyl, ethy! or benzyl, are obtained as
`mixtures of diastereomers which can be separated by chromatographic means.
`
`35
`
`Compoundsof the formula QSO,Ci and formula 1X are known or commercially available
`
`or can be made by methods well knownto those of ordinary skill in the art.
`Scheme 3 refers to the preparation of compoundsofthe formula |, wherein Z is NR' and
`is (C,-Ce)alky!,
`(Cg-Cig)aryl(C,-Cg)alkyl,
`(C.-C,)heteroaryl(C,-C.)alkyl or a group of the
`R'
`formula -(CH,),CO,R”, whereinnis 1, 3, 4, 5, or 6 and R?is (C4-Ceg)alkyl.
`
`AQUESTIVE EXHIBIT 1007
`
`AQUESTIVE EXHIBIT 1007 page 2013
`
`page 2013
`
`

`

`WO 99/52910
`
`PCT/1IB99/00503
`
`-22-
`
`Referring to Scheme 3, compounds of the formula |, wherein Z is NR’ and R'is (C,-
`(C.™-Cyo)aryi(C,-C,)alkyt,
`(C2-C,)heteroaryl(C,-C,)alkyl or a group of
`the formula
`C,)alkyl,
`-(CH,),CO2R?, wherein n is 1, 3, 4, 5, or 6 and R? is (C,-Ce)alkyl, are prepared from compounds
`of the formula XV by hydrogenolysis under an atmosphere of hydrogen in the presence of a
`catalyst in a reaction inert solvent. Suitable catalysts include 5% palladium on barium sulfate
`or 5% palladium on carbon, preferably 5% palladium on barium sulfate. Suitable solvents
`include an alcohol such as ethano!, methanol or isopropanol, preferably methanol. The
`
`to about 5 atmospheres,
`aforesaid reaction may be performed at a pressure from about 1
`preferably about 3 atmospheres. Suitable temperatures for the aforesaid reaction range from
`about 20°C (room temperature) to about 60°C, preferably the temperature may range from
`about 20°C to about 25°C (i.e. room temperature). The reaction is complete within about 0.5
`
`10
`
`15
`
`hours to about 5 hours, preferably about 3 hours.
`
`The compound of formula XV is prepared from a compound of the formula XVI by
`
`reaction with O-benzylhydroxylamine hydrochloride in the presence of a catalyst and a base
`
`in
`
`a
`
`reaction
`
`inert
`
`solvent.
`
`Suitable
`
`catalysts
`
`include
`
` (benzotriazol-1-
`
`20
`
`25
`
`yloxy)tris(dimethylamino)phosphonium hexafiuorophosphate or 1-(3-(dimethylaminopropyl)-3-
`
`ethylcarbodiimide—hydrochloride, preferably (benzotriazol-1-yloxy)tris(dimethylamino)
`
`phosphonium hexafiuorophosphate.
`Suitable bases include tertiary amines such as
`triethylamine,
`diisopropylethylamine
`or
`4-N,N-dimethylaminopyridine,
`preferably
`diisopropylethylamine. The aforesaid reaction temperature is from about 0° C to about 60°C,
`preferably about 50° C. Suitable solvents include N,N-dimethylformamide or halogenated
`solvents
`such as methylene chloride or chloroform, preferably the solvent
`is N,N-
`dimethylformamide. The reaction is conducted over a period of about 4 hours to about 48
`hours, preferably about 16 hours.
`is prepared from a compound of the formula XVII by
`The compound of formula XVI
`removal of the benzyl protecting group. Specifically, the benzyl protecting group is removed by
`hydrogenolysis using palladium or palladium on carbon in a solvent such as methanol or
`ethanol, for a period from about 30 minutes to about 48 hours, preferably 16 hours, at a
`temperature of about 20°C to about 25°C (i.e., room temperature).
`The compoundof formula XVI! is prepared from a compoundofthe formula Xil, wherein
`PG?is benzyl, by reaction with a reactive derivative of an alcohol of the formula R'OH such as
`the methanesulfonate, tosylate, chloro, bromo or iodo derivative, preferably the iodo derivative,
`in the presence of a base such as potassium carbonate or sodium hydride, preferably sodium
`hydride, and a polarsolvent, such as N,N-dimethyiformamide. The reaction mixture is stirred at
`room temperature for a time period between about 60 minutes to about 48 hours, preferably
`about 16 hours.
`
`30
`
`35
`
`40
`
`AQUESTIVE EXHIBIT 1007
`
`AQUESTIVE EXHIBIT 1007 page 2014
`
`page 2014
`
`

`

`WO 99/52910
`
`PCT/1B99/00503
`
`-23-
`
`The compounds of formula XH, wherein PG? is benzyl, are prepared according to the
`methods of Scheme 2.
`,
`Scheme4 refers to the preparation of compounds of formula I, wherein Z is >NR’, R'is
`a groupof the formula -(CH2),CO,R’(i.e. n is 2) and R? is (C,-Cg)alkyl.
`Referring to Scheme 4, compoundsof said formula | are prepared from compounds of
`the formula XVIII, wherein R? is (C,-C,)alkyl, by reaction with oxalyl chloride or thionyl chloride,
`
`10
`
`preferably oxalyl chloride, and a catalyst, preferably about 2% of N,N-dimethylformamide, in an
`
`inert solvent, such as methylene chioride, to form an in situ acid chloride that is subsequently
`
`reacted with O-trimethylsilylhydroxytamine in the presence of a base, such as pyridine, 4-N,N-
`
`dimethylaminopyridine or triethylamine, preferably pyridine. The reaction is performed at a
`
`15
`
`temperature of about 22°C (i.e., room temperature) for about 1
`
`to about 12 hours, preferably
`
`about 1 hour.
`
`Compounds of the formula XVIII, wherein R?is (C,-Ce,)alky!, can be prepared from
`compounds of the formula XIX, wherein R? is (C,-C,)alkyl, by reduction in a polar solvent.
`
`Suitable reducing agents include hydrogen over palladium and hydrogen over palladium on
`
`20
`
`carbon, preferably hydrogen over palladium on carbon. Suitable solvents include methanol,
`
`ethanol and isopropanol, preferably ethanol.
`
`The aforesaid reaction is performed at a
`
`temperature of about 22°C (i.e., room temperature) for a period of 1 to 7 days, preferably about 2
`
`days.
`
`25
`
`Compounds of the formula XIX, wherein R? is (C,-Ce)alkyl, can be prepared from
`compoundsof the formula Xil, wherein PG?is benzyl, by Michael addition of a propiolate ester
`and a basein a polar solvent. Suitable propiolates are of the formula H-C=C-CO,R?, wherein R?
`
`is (C,-C,)alkyl. Suitable bases include tetrabutylammonium fluoride, potassium carbonate, and
`
`cesium carbonate,
`
`preferably tetrabutylammonium _fiuoride.
`
`Suitable
`
`solvents
`
`include
`
`tetrahydrofuran, acetonitrile, tert-butanol and N,N-dimethylformamide, preferably tetrahydrofuran.
`
`30
`
`The aforesaid reaction is performed at a temperature of about -10°C to about 60°C, preferably
`
`35
`
`ranging between 0°C and about 22°C (i.e., room temperature). The compounds of formula XIX
`
`are obtained as mixtures of geometric isomers about the olefinic double bond; separation of the
`
`isomers is not necessary.
`Compoundsofthe formula XII, wherein PG? is benzyl, can be prepared according to the
`methods of Scheme 2.
`is a group of the formula
`Compounds of said formula |, wherein Z is >NR', R'
`-(CH,),CO,R? , n is 1
`to 6 and R® is hydrogen are prepared from compoundsof formula |,
`wherein Z is >NR’, R'is a groupof the formula -(CH,),CO,R?, n is 1 to 6 and R?is (Cy-C,)alkyl,
`
`by saponification using a base such as sodium hydroxide in a protic solvent such as ethanol,
`
`40
`
`methanol!or water or a mixture such as water and ethanol, water and toluene, or water and THF.
`
`AQUESTIVE EXHIBIT 1007
`
`AQUESTIVE EXHIBIT 1007 page 2015
`
`page 2015
`
`

`

`WO 99/52910
`
`PCT/IB99/00503
`
`-24-
`
`The preferred solvent system is water and ethanol. The reaction is conducted for a period of 30
`minutes to 24 hours, preferably about 2 hours.
`The compounds of the formula | which are basic in nature are capable of forming a
`wide variety of different salts with various inorganic and organic acids. Although such salts
`must be pharmaceutically acceptable for administration to animals,
`it
`is often desirable in
`practice to initially isolate a compound of the formula | from the reaction mixture as a
`
`pharmaceutically unacceptable salt and then simply convert the latter back to the free base
`compoundbytreatmentwith an alkaline reagent, and subsequently convert the free base to a
`pharmaceutically acceptable acid addition salt.
`The acid addition salts of
`the base
`
`compounds of this invention are readily prepared by treating the base compound with a
`substantially equivalent amount of the chosen mineral or organic acid in an aqueous solvent
`medium or
`in a suitable organic solvent such as methanol or ethanol. Upon careful
`evaporation of the solvent, the desired solid salt is obtained.
`
`The acids which are used to prepare the pharmaceutically acceptable acid addition
`salts of the base compoundsof this invention are those which form non-toxic acid addition
`Salts,
`i.e., salts containing pharmacologically acceptable anions, such as hydrochloride,
`hydrobromide, hydroiodide,nitrate, sulfate or bisulfate, phosphate or acid phosphate, acetate,
`lactate,citrate or acid citrate, tartrate or bitartrate, succinate, maleate, fumarate, gluconate,
`saccharate, benzoate, methanesulfonate and pamoate [i.e., 1,1'-methylene-bis-(2-hydroxy-3-
`naphthoate)] salts.
`
`Those compounds of the formula | which are also acidic in nature, are capable of
`forming base salts with various pharmacologically acceptable cations. Examples of such salts
`include the alkali metal or alkaline-earth meta! salts and particularly,
`the sodium and
`potassium salts. These salts are all prepared by conventional techniques. The chemical
`bases which are used as reagents to prepare the pharmaceutically acceptable base salts of
`this invention are those which form non-toxic base salts with the herein described acidic
`
`compounds of formula |. These non-toxic base salts include those derived from such
`
`pharmacologically acceptable cations as sodium, potassium, calcium and magnesium,etc.
`These salts can easily be prepared by treating the corresponding acidic compounds with an
`aqueous solution containing the desired pharmacologically acceptable cations, and then
`evaporating the resulting solution to dryness, preferably under
`reduced pressure.
`Alternatively, they may also be prepared by mixing lower alkanolic solutions of the acidic
`compoundsand the desired alkali metal alkoxide together, and then evaporating the resulting
`solution to dryness in the same manneras before.
`In either case, stoichiometric quantities of
`reagents are preferably employed in order to ensure completeness of reaction and maximum
`productyields.
`
`10
`
`15
`
`20
`
`25
`
`30
`
`35
`
`40
`
`AQUESTIVE EXHIBIT 1007
`
`AQUESTIVE EXHIBIT 1007 page 2016
`
`page 2016
`
`

`

`WO 99/52910
`
`PCT/1B99/00503
`
`-25-
`
`The ability of the compounds of formula ! or their pharmaceutically acceptable salts
`
`(hereinafter also referred to as the compounds of
`
`the present
`
`invention)
`
`to inhibit
`
`metalloproteinases or mammalian reprolysin and, consequently, demonstrate their effectiveness
`
`for treating diseases characterized by metalloproteinase or the production of tumor necrosis
`
`factor is shown by the following in vitro assay tests.
`
`10
`
`Biological Assay
`inhibition of Human Collagenase (MMP-1)
`
`Human recombinant collagenase is activated with trypsin. The amount of trypsin is
`
`optimized for each lot of collagenase-1 but a typical reaction uses the following ratio: 5 1g trypsin
`
`per 100 ng of collagenase. The trypsin and collagenase are incubated at room temperature for
`
`15
`
`10 minutes then a five fold excess (50 mg/10 mg trypsin) of soybean trypsin inhibitor is added.
`
`Stock solutions (10 mM)of inhibitors are made up in dimethytsulfoxide and then diluted
`
`using the following scheme:
`
`
`
`10 mM ---—> 120 pM ----—-> 12 p