throbber
WO 90/05719
`
`oo
`
`PCT/GB89/01399
`
`50
`
`(4-Hydroxy-2R-isobuty1-38-(2,4-dimethylphenylthio-
`1
`2 methyl) succinyl ]~—L-phenylalanine-N-methylamide (1.89,
`3
`3.7 mmol) and HOBT (0.67g, 12 mmol) were dissolved in
`4
`1:1 DCM/DMF and the mixture cooled to 0°C before adding
`S WSDCI (0.86q, 4.5mmol) and NMM (0.45g, 4.5mmol). The
`6 mixture was stirred at o°c for ih to ensure complete
`7
`formation of
`the activated ester. Hydroxylamine
`8
`hydrochloride (0.39g, 5.6mmol) and NMM (0.56g, 5.6mmol)
`9 were dissolved in DMF
`then this mixture was added
`10
`dropwise to the cooled solution of the activated ester.
`4 After Ih the reaction was poured into ether/water (1:1)
`12 whereupon the desired product precipitated as white
`13° crystals. These were collected by filtration, further
`14 washed with ether and water,
`then dried under vacuum at
`15
`50°C. This material was repeatedly recrystallised from
`16 methanol/water
`(1:1)
`to remove a trace of the minor
`17 @iastereomer (1.08g, 2.2mmol, 58%).
`18
`.
`
`19° mip. 226°C (dec.)
`20
`
`21 Analysis calculated for Co7H37N30,5
`22 Requires: C64.90 H7.46 N8.41
`23°
`Found:
`C65.15 H7.48 N8.40
`24
`.
`25 delta, (250MHz, Dg-DMSO) 8.83 (1H, s, NHOH), 8.32 (1H,
`26
`d,
`Jd = 8Hz, CONH), 7.85 (1H,
`dad, J = 6Hz, CONHMe), 7.30
`27
`- 6.71 (9H, m, aromatic H), 4.56 (iH, mn, CHCH5Ph), 2.91
`28
`(4H, dd,
`J = 14,4Hz, CHCH,Ph),
`2.76 (1H,
`dd,
`J =
`29
`14,10Hz, CHCH)Ph), 2.57 (3H, d, J = 4.5Hz, NHCH), 2.53
`30,
`~ 2.38 (2H, m), 2.23 (3H,
`5s, CoH5(CH3)2), 2.13 (3H, s,
`* CgHs(CH3), 1-30 (2H, m), 0.89 (1H, m, CH2CH(CH3)),
`0.81 (3H, d, J = 6Hz, CH(CH3)5), and 0.74 (3H, d, J =
`-
`6Hz, CH(CH3)5)- ——
`-
`
`33
`
`*
`a.
`
`AQUESTIVE EXHIBIT 1004
`
`AQUESTIVE EXHIBIT 1004 page 0701
`
`page 0701
`
`

`

`WO 90/05719
`
`_ PCT/GB89/01399
`
`51
`
`
`
`CONHOH
`
`O
`
`Example 15
`
`7
`
`2 3 4 5 6 7 8
`
`A
`
`O
`
`NHMe
`
`H
`
`9
`10
`11
`Ph
`12
`[4 (N-Hydroxyamino—2R-isobuty1l-3S8- (acetylthiomethyl)
`13
`succinylj}-L-phenylalanine-N-methylamide (1.0g, 2.4
`14
`15 mmol) was dissolved in 750ml methanol and 350ml pH 7
`16
`buffer added. Left
`to stand overnight and solvent
`7 removed in vacuo to 2/3 volume,left to crystallise for
`18
`3 further two hours. Filtered and dried to give 0.87g
`19
`off-white crystals
`20
`
`HN
`
`H
`
`21 Analysis calculated for C3gHs6Ne0gSn-1.9H20
`22 Requires: C55.34 H6.93 N9.88
`23
`Found:
`C55.44 H7.32 N10.21
`24
`
`25
`26
`
`27
`28
`29
`
`30
`31
`
`32
`
`33
`
`Example 16
`
`[4=(N-Hydroxyamino) -2R-isobuty1-38~- (3-bromophenyl-
`thiomethyl) succinyl ]-L-phenylalanine-N-methylamide
`Ph
`
`CONHOH Br
`
`AY
`
`.
`
`NHMe
`
`AQUESTIVE EXHIBIT 1004 page 0702
`
`AQUESTIVE EXHIBIT 1004 page 0702
`
`

`

`WO 90/05719
`
`PCT/GB89/01399
`
`onyaDthmWwWh&
`
`‘oO
`
`17
`
`12
`
`13
`
`714
`
`15
`
`16
`
`17
`
`18
`
`19
`
`20
`
`21
`
`22
`
`23
`
`24
`
`25
`26
`
`27
`
`28
`
`29
`
`30
`
`31
`
`32
`
`33
`
`52
`
`Prepared by the method described in example 1g to give
`material with the following characteristics.
`
`m.p. 225 -229°¢
`
`[alpha]p = -164.8°
`
`Analysis calculated for Cz H35BrN30,8
`Requires: C54.40 H5.89 N7.40
`Found:
`C54.54 H5.86 N7.63_
`
`(250MHz, Dg-DMSO) 8.83 (1H, s, NHOH), 8.35 (1H,
`delta,
`d, J = 8Hz, CONH), 7.90 (1H, q, J = 6Hz, CONHMe), 7.35
`-- 6.87 (9H, m, aromatic H), 4.64 (1H, m, CHCH5Ph), 2.94
`(1H, dd, J = 14,4Hz, CHCH,Ph), 2.76 (1H, t, J = 13Hz,
`‘CHCH,Ph) 2.60 (3H, d, J = 5Hz, NHCH3), 2.55 - 2.35 (2H,
`m, CHoS), 2.15 (1H, t, J = 10Hz, CHCO), 2.01 (1H, a, J
`-= 11.5Hz,
`CHCO),
`1.37 (2H, m),
`0.88 (1H,
`n,
`CH>CH(CH3)5), 0-81 (3H, d, J = 6Hz, CH(CH3)5), and 0.74
`(3H, G,Jd = 6HZ,CH(CH3)>5)..
`
`(63.9MHz, D,-DMSO) 173.0, 171.0, 168.8, 139.8,
`delta,
`138.0, 130.5, 129.0, 128.5, 127.5, 125.8, 125.5, 54.2,
`46.0, 45.5, 38.0, 31.5, 25.5, 25.2, 24.7, and 21.0.
`
`Example 17
`
`[4-(N-Hydroxyamino) -2R-isobuty1~-3S- (3-chlorophenylthio-
`methyl) succinylj-L-phenylalanine-N-methylamide
`Bh
`
`.
`
`AQUESTIVE EXHIBIT 1004=page 0703
`
`AQUESTIVE EXHIBIT 1004 page 0703
`
`

`

`WO 90/05719
`
`.
`
`PCT/GB89/01399
`
`53
`
`Prepared by the method described in example 1g to give
`1
`2 material with the following characteristics.
`
`m.p. 231-234°C
`
`[alpha], = -96.5°
`
`3 4
`
`5 6
`
`7 8
`
`Analysis calculated for C5°H32C1N30,5
`9 Requires: C59.34 H6.37 N8.30
`10
`Found:
`C59.51 H6.43 N8.24
`
`11
`
`(250MHZ, Dg-DMSO) 8.85 (1H, s, NHOH), 8.37 (1H,
`12 delta,
`13.
`da, J = 8.5Hz, CONH), 7.90 (1H, m, CONHMe), 7.30 -— 6.88
`14
`(9H, m, aromatic H), 4-66 (1H, m, CHCH,Ph), 2.96 (1H,
`15
`bd,
`J = 14Hz,
`CHCH»Ph),
`2.76 (1H, bt,
`J = 13Hz,
`16
`CHCH,Ph) 2.60 (3H, d, J = 5Hz, NHCH3), 2.55 - 2.40 (2H,
`17
`m, CH5S), 2-16 (1H, m, CHCO), 2-01 ( 1H, d, J = 14Hz,
`18
`CHCO), 1.37 (2H, m), 0.91 (1H, m, CH5ZCH(CH3)5), 0.81.
`19
`(3H, d,
`J = 6Hz, CH(CH3)5),
`and 0.74 (3H,
`da,
`J =
`20
`6Hz,CH(CH;)>)-
`21
`
`(63.9MHz, Dg-DMSO) 172.7, 171.6, 168.1, 139.2,
`deltag
`138.1, 130.3, 129.2, 127.9, 126.2, 125.9, 125.5, 125.0,
`54.1, 46.3, 45.8, 37.8, 32.0, 25.7,
`25.2,
`24.2,
`and
`21.7.
`
`22
`23
`24
`25
`
`26
`
`27
`
`28
`
`29
`
`30
`
`31
`
`32
`
`33
`
`AQUESTIVE EXHIBIT 1004
`
`AQUESTIVE EXHIBIT 1004 page 0704
`
`page 0704
`
`

`

`WO 90/05719.
`
`PCT/GB89/01399
`
`54
`
`, Example18
`
`[4-(N-Hydroxyamino)-2R-isobuty1l-3S-(3-
`3
`4 methylphenylthiomethyl) succinyl]-L-phenylalanine-N-
`5 methylamide
`6
`.
`:
`
`Ph
`NHMe
`
`
`
`9
`10
`
`11
`
`12
`
`13
`
`Me.
`
`CONHOH
`
`14
`15 Prepared by the method described in example ig to give
`16 Material with the following characteristics.
`17
`18 Analysis calculated for Co 6H35N3045
`49 Requires: C64.30 H7.26 N8.65
`29
`Found:
`C63.81 H7.21 N8.48
`21
`.
`22- delta,
`(250MHz, D,g-DMSO) 8.83 (1H, s, NHOH), 8.35 (1H,
`23 d, J = 8.5Hz, CONH), 7.86 (1H, m, CONHMe), 7.28 - 6.77
`24
`(9H, m, aromatic H), 4.66 (1H, m, CHCH5Ph), 2.96 (1H,
`25
`ad, J = 14,4Hz, CHCH5Ph), 2.80 (1H, bt, J = 13Hz,
`26
`CHCH>5Ph ) 2.59 (3H, d, J = 5Hz, NHCH3), 2.55 - 2.37 (2H,
`27 ™, CHS), 2.16 (2H, m, 2xCHCO), 1.38 (2H, m), 0.91 (1H,
`2g ™, CH)CH(CH3)5), 0.81 (3H, d, J = 6Hz, CH(CH3)5), and
`29
`0.74 (3H, d, J = 6Hz, CH(CH3)>).
`
`30 |
`31
`32
`33
`
`.
`
`:
`oo
`
`.
`
`.
`
`.
`
`*
`
`.
`
`AQUESTIVE EXHIBIT 1004=page 0705
`
`AQUESTIVE EXHIBIT 1004 page 0705
`
`

`

`WO 90/05719
`
`PCT/GB89/01399
`
`onanaUFWHY
`
`
`
`WWWWSnYNYNNNNNNNN@-BBBB|amwrorWNOOWNDUFwWNYAOWDWBATHNBwWHNY|OC
`
`55
`
`Example 19
`
`[4-(N-Hydroxyamino)-2R-isobuty1l-3S-(4-(N-acetyl)-
`aminophenylthiomethyl)succinyl]-L-phenylalanine-N-
`methylamide.
`-
`
`Ph
`
`wt NHMe
`
`CONHOH
`
`ACNH
`
`A)
`
`[2R-isobuty1-3S-(4-aminophenylthiomethyl )succinyl}-
`L-phenylalanine -N-methylamide.
`
`Prepared by the method described in example 1f to give
`material with the following characteristics.
`
`(250MHz, Dg-DMSO) 8.27 (1H, d, J = 8.5Hz, CONH),
`deltay,
`7.81
`(1H, m, CONHMe), 7.30 - 7.00 (5H, m, phenyl H),
`6.86 (2H, d, J = 8.5Hz, aromatic H), 6.45 (2H, d, J =
`8.5Hz, aromatic H), 5.25 (1H, bs, COzH), 4.48 (1H, m,
`CHCH5Ph), 2.91 (1H, dd, J = 14,4Hz, CHCH>Ph), 2.88 (1H,
`dd, J = 14,10Hz, CHCH,Ph) 2.56 (3H, d, J = 5Hz, NHCH3),
`2.43 - 2.24 (3H, m, CHjS and CHCO), 2.03 (1H, d, @ =
`10Hz, CHCO), 1.41 (1H, t, J = 11Hz, CHyCH(CH3)5), 1.26
`
`J = 6Hz, CH(CH3)5),
`(3H, d,
`CH(CH3))-
`
`and 0.74 (3H, d,
`
`J=6Hz,
`
`AQUESTIVE EXHIBIT 1004 page 0706
`
`AQUESTIVE EXHIBIT 1004 page 0706
`
`

`

`~ WO 90/05719
`
`oo
`
`56
`
`PCT/GB89/01399
`
`t
`
`.
`
`.
`
`4
`Q
`
`[2R-isobutyl-38-(4-(N-acetyl)aminophenyl-thio-
`8B)
`=Mmethyl)- succinyl ]-Lphenylalanine-N-methylamide.
`
`3 4
`
`The product from above (350mg, 0.74 mmol) was dissolved
`in DCM (5 ml) cooled in an ice bath then triethylamine
`-s5
`(75mg, 0.74 mmol), DMAP
`(91mg, 7.4 mmol) and finally
`6
`7 acetic anhydride (83mg, 8.2 mmol) were added and the
`8
`solution stirred at RT for 90 minutes.
`The mixture was.
`9 partitioned between ethyl acetate and citric acid then
`10
`the organic layer washed with water and finally dried
`11.
`over Magnesium sulphate. Solvent removal gave the crude
`12 product as pale yellow crystals (160mg, 0.31 mmol,
`13 42%).
`C
`
`;
`.
`14
`15 deltay (250MHz, Dg-DMSO) 9.94 (1H, s, CO,H), 8.34 (1H,
`16
`Gr J = 8.5Hz, CONH), 7.90 (1H, m, CONHMe), 7.46 (2H, d,
`17
`9 = 8.5Hz, aromatic H) 7.30 ~ 7.00 (5H, m, phenyl H),
`4g
`6-96 (2H, a, J = 8.5Hz, aromatic H), 4.57 (1H, m,
`19
`CHCH2Ph), 2.91 (1H, dd, J = 14,4Hz, CHCH,Ph), 2.88 (1H,
`290 bt, J = 13Hz, CHCH)Ph), 2.58(3H, d, J = 5Hz, NHCH3),
`21
`2-43 - 2.16 (3H, m, CH5S and CHCO), 2.10 (1H, d, 3 =
`22
`14Hz, CHCO), 1.35 (1H, t, J = 14Hz, CHyCH(CH3)5), 1.26
`23
`(1H,
`4m, CHoCH(CH3)5), 0.86 (1H, m, CHyCH(CH3)5), 0.81
`gq
`(3H, G, J = 6Hz,° CH(CH3)2),
`and 0.74 (3H, 4, J =
`55 6Hz,CH(CH3)>).
`mo
`a
`26
`SO
`27 _C)
`[ 4-(N-Hydroxyamino) -2R-isobuty1-3S-(4- (N-acetyl) -
`28
`aminophenylthiomethy1 ) succinyl ]-L—phenylalanine-Nn-
`29
`methylamide.
`
`30
`.
`31 Prepared by the method described in example 1g to give
`32 material with the following characteristics.
`
`33
`
`~
`
`AQUESTIVE EXHIBIT 1004 page 0707
`AQUESTIVE EXHIBIT 1004
`page 0707
`
`

`

`WO 90/05719
`
`PCT/GB89/01399
`
`57
`
`m.p. 201 -202°C (dec.)
`
`[alpha]p = -7.5° (c=1.0, methanol)
`
`(250MHz, Dg-DMSO) 9.90 (1H, s, NHOH), 8.82 (1H,
`delta,
`s, NHOH), 8.30 (1H, d, J = 8.5Hz, CONH), 7.85 (1H, m,
`CONHMe), 7.45 (2H, d, J = 8.5Hz, aromatic H), 7.28 -
`6.94 (5H, m, phenyl H), 6.90 (2H,
`da,
`3 = 8.5Hz,
`aromatic H), 4.66 (1H, m, CHCH,Ph), 2.90 (1H, dd, J =
`14,4Hz, CHCH>Ph), 2.76 (1H, bt,
`J = 13Hz, CHCH5Ph),
`2.50 (3H, d, J = 5Hz, NHCH,), 2.49 - 2.35 (2H, m,
`CH>S), 2.14 (1H, m, CHCO), 2.03 (4H,
`s + m, COCH3 and
`CHCO), 1.35 (2H, m), 0.86 (1H, m, CHCH(CH3)5), 0.81
`(3H, d, J-= 6Hz, CH(CH3)5), and 0.74 (3H, d, J = 6Hz,
`CH(CH3)>).
`
`Example 20
`
`[ 4-(N-Hydroxyamino) -2R-isobutyl-3S-phenylsulfinyl-
`methylsuccinyl]-L-phenylalanine-N-methylamide,
`
`
`
`[4-(N-Hydroxyamino) -2R-isobuty1-35-phenylthiomethyl-
`succinyl]-L-phenylalanine-N-methylamide (250mg,
`0.53mmol) was dissolved in methanol
`(50 ml) and meta-
`
`oannmPFWNY
`
`Ne}
`
`41
`
`12
`
`13
`
`14
`
`15
`
`16
`
`17
`
`18
`
`19
`
`20
`
`21
`
`22
`
`23
`
`24
`
`25
`
`26
`
`27
`
`28
`
`29
`
`30
`
`31
`
`32
`
`33
`
`AQUESTIVE EXHIBIT 1004 page 0708
`AQUESTIVE EXHIBIT 1004=page 0708
`
`

`

`WO 90/05719
`
`PCT/GB89/01399
`
`58
`
`0.58 mmol) was added.
`chloroperbenzoic acid (100mg,
`After stirring for th at
`room temperature ether was
`added and the mixture filtered.
`Solvent removal gave
`the crude white solid which was
`recrystallised from
`methanol / water then slurried in ether to remove final
`traces of meta-chlorobenzoic acid to give the desired
`material
`(70 mg, 0.014 mmol, 27%).
`
`+
`
`x
`
`m.p. 186 -1889C
`
`[alphalp = -13.6° (c=0.5, methanol)
`
`Analysis caiculated for C95H33N305S.0.5H50
`Requires: C60.46 H6.90 N8.46
`
`Found:.
`
`C60.58 H6.69 N8.29
`
`delta, (250MHz, Dg-DMSO, mixture of diastereomers) 9.04
`+ 8.93 (1H, 2xs, NHOH), 8.29 + 8.16 (1H, 2xd, J = 8.5
`Hz, CONH), 7.79 (1H, m, CONHMe), 7.90 - 7.40 (8H, m,
`aromatic H), 7.06 + 6.82 (2H,
`2xm, SO-Aromatic), 4.37
`(1H, m, CHCHPh), 2.93 - 2.58 (3H, m, containing
`CHCH>Ph), 2.52 (3H, m, NHCH3), 2.49 + 2.37 (1H, 2xm),
`1.49 - 1.25 (2H, m, CHjCH(CH3)5 and CH2CH(CH3)5), 0.95
`(1H, m, CHCH(CH3) ), 0.81 (3H, d, J = 6Hz, CH(CH3)5),
`and 0.74 (3H, d, J=6Hz, CH(CH3)5).
`
`(63.9MHz, De-DMSO, mixture of diastereomers)
`delta,
`172.2, 171.4, 171.3, 167.7,144.5, 138.0, 137.9, 131.3,
`130.9, 129.6, 129.3, 129.1, 128.8, 128.3, 127.8, 126.5,
`126.2, 124.3, 123.6, 59.8, 58.1, 54.3, 54.0, 46.2,
`45.8, 41.6, 40.9, 37.6, 37.4, 25.6, 25.0, 24.3, 24.2,
`21.7, and 21.6.
`~
`
`onam&®WN
`
`
`
`WnF-oOUODANTHONBWNYHPTOWANAUPWNOCWO
`
`AQUESTIVE EXHIBIT 1004
`
`AQUESTIVE EXHIBIT 1004 page 0709
`
`page 0709
`
`

`

`WO 90/05719 |
`
`~
`
` PCT/GB89/01399
`
`59
`
`Example 21
`
`[4-(N-Hydroxyamino) -2R-isobuty1-3S-phenylsulfonyl-
`methylsuccinyl ]-L-phenylalanine-N-methylamide.
`
`
`
`[4-(N-Hydroxyamino)-2R-isobutyl1-3S8-phenylthiomethyl-
`
`(50mg,
`succinyl]-L-phenylalanine-N-~methylamide
`0.11mmol) was dissolved in methanol
`(12 ml) and meta-
`chloroperbenzoic acid (40mg, 0.23 mmol) was added.
`After stirring for 3h at
`room temperature ether was
`
`added and the mixture filtered.
`
`Solvent
`
`removal gave
`
`the crude white solid which was slurried in ether to
`
`remove final traces of meta-chlorobenzoic acid to give
`
`the desired material.
`
`mop. 228 - 231°C
`
`[alpha]lp = 16.8° (c=0.5, methanol)
`
`Analysis calculated for Cy5H33N30,¢S.0.3H»50
`Requires: C58.99 H6.65 N&.25
`
`Found:
`
`C58.92 H6.51 N8.05
`
`deltay (250MHz, Dg-DMSO) 8.66 (1H, s, NHOH), 8.25 (1H,
`Gd, J = 8.5 Hz, CONH), 7.83 (1H, m, CONHMe), 7.75 - 7.50
`(5H, m, aromatic H), 7.30 7.05 (5H, m,
`aromatic H),
`
`onyDOkWdh+
`
`11
`
`12
`
`13
`
`14
`
`15
`
`16
`
`17
`
`18
`
`19
`
`20
`
`21
`
`22
`
`23
`
`24
`
`25
`
`26
`
`27
`
`28
`
`29
`
`30
`
`31
`
`32
`
`33
`
`AQUESTIVE EXHIBIT 1004 page 0710
`AQUESTIVE EXHIBIT 1004 page 0710
`
`

`

`WO90/03719
`
`PCT/GB89/01399
`
`60
`
`oITHOeAWDdA
`
`WWWWNNNNNNNNNNHRPBSBBABBPBeeAreaWnsFHOUODAKHUNBRwWNYNHWOHMMNDUbtWN|OO
`
`4.36 (1H, m, CHCH,Ph), 2.86 (1H, dd, J = 14,5 Hz,
`CHCH»Ph), 2.75 (1H, dd, J = 14,10 Hz, CHCH5Ph), 2.54
`(3H,
`ad, J = 4.5 Hz, NHCH3), 2.54 (2H, m), 1.30 (2H, m,
`CH»CH(CH3)5 and. CH5CH(CH3)5),
`0.86 (1H,
`Om,
`CH5CH(CH3)5), 0.75 (3H, d, J = 6Hz, CH(CH3)5), and 0.71
`(3H, d, J = 6Hz,CH(CH3)5).
`,
`
`Example 22
`
`[4-( N-Hydroxyamino)-2 R-is obutyl-3 S-
`thiophenylsulphinylmethyl-succinyl] -L-~phenylalanine-N-
`methylamide
`
`
`
`[4-(N-Hydroxyamino) -2R-isobuty1-3S-thiophenylthio-
`methyl-succinyl]-L-phenylalanine-N-methylamide (50mg,
`O.11mmol) was
`treated as described in example 21
`to
`yield the title compound (16mg, 0.03 mmol,
`29%) as a
`mixture of diastereomer with the following
`characteristics:
`
`m.p. 195 -197°C (dec.)
`
`Analysis calculated for C73H34N30.S5.0.5H50
`Requires: C54.96 H6.42 N8.36
`Found:
`.C54.91 H6.23 N8.23
`
`AQUESTIVE EXHIBIT 1004 page 0711
`. AQUESTIVE EXHIBIT 1004
`
`page 0711
`
`

`

`WO 90/05719
`
`PCT/GB89/01399
`
`61
`
`deltay (250MHz, Dg-DMSO, mixture of diastereomers) 9.04
`+ 8.96 (1H, 2xs, NHOH), 8.34 + 8.29 (1H, 2xd, J = 8.5
`3 Hz, CONH), 8.02 + 7.98 (1H, 2xm, CONHMe), 7.81 (1H, bs,
`4
`thiophene-H), 7.42 (1H, s, thiophene-H), 7.25 - 7.15
`5
`(5H, m, phenyl), 7.03 (1H, bs,
`thiophene-H), 4.43 (1H,
`6 m, CHCHsPh), 3.0 - 2.6 (4H, m,
`containing CHCH>Ph),
`7
`2.52 (7H, m, containing NHCH3), 2.05 (1H, m), 1.6 ~ 1.2
`8
`(2H, m, CH5CH(CH3)2 and CHaCH(CH3)9), 0.87 (1H,
`m™,
`9 CH5CH(CHz)5), and 0.85 - 0.71 (6H, m, CH(CH3)>).
`10
`11 Example23
`
`1 2
`
`12
`[4-(N-Hydroxyamino)-2R-isobuty1-3S-
`13
`thiophenylsulphonylmethyl-succinyl]
`-L-phenylalanine-N-
`44
`145 methylamide.
`
`16
`
`17
`
`18
`
`19
`
`20
`
`21
`
`22
`
`23
`
`
`
`24
`[4-(N-Hydroxyamino) -2R-isobutyl-3S-thiophenylthio-
`25
`26 methyl-succinyl]-L-phenylalanine-N-methylamide (75mg,
`27
` 0.16mmol) was
`treated as Gescribed in example
`22
`to
`2g yield the title compound (40mg, 0.08 mmol,
`49%) with
`a9
`the following characteristics:
`
`30
`31. Mp. 215 - 216°C
`32
`33 Analysis calculated for Cy3H31N30¢S5
`
`AQUESTIVE EXHIBIT 1004=page 0712
`
`AQUESTIVE EXHIBIT 1004 page 0712
`
`

`

`WO 90/05719
`
`|
`
`|
`
`|
`
`62
`
`_ PCT/GB89/01399
`
`Requires: C54.21 H6.13 N8.24
`Found:
`C54.07 H6.19 N8.04
`
`(250MHz, D,g-DMSO) 887 (1H, s, NHOH), 8.25 (1H,
`delta,
`d, J = 8.5 Hz, CONH), 8.09 (1H, d,
`J = 4.7 Haz,
`thiophene-H), 7.83 (1H, m, CONHMe), 7.53 (1H, d, J = 3.
`Hz,
`thiophene H), 7.25 - 7.12 (6H, m, phenyl and
`thiophene-H), 4.36 (1H,-m, CHCH5Ph), 3.38 (1H, dd, 7 =
`14,11 Hz, SCH»), 2.87 (1H, dd, J = 14,5 Hz, CHCHPh),
`“2.75 (1H, dd, J = 14,10 Hz, CHCHaPh), 2.70 - 2.36 (6H,
`m, containing NHCH3;), 1.20 (2H, m, CHaCH(CH3)5 and
`CHaCH(CH3)), 0.89 (1H,m, CH5CH(CH3)>5
`), and 0.75 (6H,
`m, CH(CH3)5).
`
`(63.9MHz, Dg-DMSO) 172.0, 171.2, 166.5, 140.0,
`deltag
`138.0, 135.4, 134.6, 129.0, 128.4, 128.2, 126.6, 54.3,
`45.6, 37.5, 25.6, 25.0,.24.2, and 21.7.
`a
`
`Example 24
`
`[4-(N-Hydroxyamino)-~2R-isobutyl-3S-phenylsulfonyl-
`methylsuccinyl]-L-phenylalanine-N-methylamide sodium
`salt.
`
`
`
`onamPpwWh—
`
`WWwWwNnNNNNNNNNNDN|BABABAAwmmeotNy-OMYMONDUBwWNBADOHMHANIHUPWN~~CO
`
`33
`
`[4-(N-Hydroxyamino)-2R-isobuty1-3s-phenylsulfonyl-
`
`AQUESTIVE EXHIBIT 1004=page 0713
`
`AQUESTIVE EXHIBIT 1004 page 0713
`
`

`

`WO90/05719
`
`PCT/GB89/01399
`
`63
`
`orDm&FWHY-
`
`WwWWWwnNNdNSNYNYNNNNNNDSF&BFABABABASs|AWNAOODAKDHBWNYADOHDAIHinbhWNH-OOLO
`
`methylsuccinyl]-L-phenylalanine-N-methylamide (50mg,
`O0.1mmol) was dissolved in methanol
`(100ml)
`and sodium
`hydroxide solution (0.1M,
`1.0ml)
`added to give a
`homogeneous solution.
`The methanol was
`removed under
`reduced pressure then the residual
`aqueous
`solution
`freeze dried to give the title compound (40mg).
`
`(250MHz, De-DMSO) 8.66 (1H, s, NHOH), 8.25 (1H,
`delta,
`d, J = 8.5 Hz, CONH), 7.83 (1H, m, CONHMe), 7.75 - 7.50
`(5H, m, aromatic H), 7.30 7.05 (5H, m, aromatic H),
`4.36 (1H, m, CHCH5Ph), 2.86 (1H, dd,
`J = 14,5 Hz,
`CHCHoPh), 2.75 (1H, dd,
`J = 14,10 Hz, CHCHsPh), 2.54
`(3H,
`Gd,
`J=4.5 Hz, NHCH3), 2.54 (2H, m), 1.30 (2H, m,
`CH,CH(CH3)5
`and CHyCH(CH3)9)-
`0.86
`(1H,
`m,
`CH5CH(CH3)9), 0.75 (3H, d, J = 6Hz, CH(CH3)9), and 0.71
`(3H, d, J = 6Hz, CH(CH3)>5)-
`
`Example 25
`
`[4-(N-Hydroxyamino)-2R-isobuty1-3S-(4-(isobutyloxy-
`carbonylamino) phenyl)thiomethyl-succinyl]-L-phenyl-
`alanine-N-methylamide
`
`
`
`oO
`
`a)
`
`[4-Hydroxy-2R-isobuty1~-3S-(4-aminophenyl )thio-
`
`AQUESTIVE EXHIBIT 1004
`
`AQUESTIVE EXHIBIT 1004 page 0714
`
`page 0714
`
`

`

`=
`
`¥
`
`WO 90/05719
`
`a
`
`PCT/GB89/01399
`
`64
`
`4 methylsuccinyl]-L-phenylalanine-N-methylamide was
`2 prepared by the method described in example if to give
`3 a compound with the following characteristics.
`4
`|
`-
`|
`|
`5 delta,
`(250MHz, De-DMSO) 8.26 (1H, d,
`JF = 8.5 Hz,
`CONH), 7.81 (1H, m, CONHMe), 7.27 - 7.15 (5H, m, phenyl
`H), 6.85 (2H, d, J = 8.5Hz, aromatic H), 6.46 (2H, d, J
`= 8.5Hz, aromatic H), 5.2 (1H, bs, CO,H), 4.48 (1H, m,~
`9 CHCH,Ph), 2.90 (1H, dd, J = 13.5,4.3 Hz, CHCH5Ph), 2.75
`10
`(1H, dd, J = 13.6, 10 Hz, CHCH5Ph), 2.56 (3H, d,
`gc =
`11 4.5 Hz, NHCH3), 2.50-+ 2.25 (3H, m), 2.03 (1H, d, J =
`12
`10 Hz),
`1.41 (1H, m, CH,CH(CH3)5),
`1.26 (1H, n,
`13. CH)CH(CH3)9), 0.86 (1H, m, CH5CH(CH3)>), 0.75 (3H, a, J
`14
`= 6Hz, CH(CH3)5), and 0.71 (3H, d, J = 6Hz,CH(CH3)>5).
`15 OB
`-
`.
`16 Db) N,N-Dimethyliglycine (100mg, 0.97 mmol). was stirred
`17.
`in dry THF
`(50m1) and triethylamine (108mg,
`1.1mmol)
`18
`and isobutylchloroformate (146mg, 1.1mmol) were added.
`19 After th the product from example 26a (500mg, 1.1mmol)
`20 was addedand the mixture stirred for a further th. The
`21
`reaction was worked up by partitioning between citric
`22
`acid and ethyl acetate, drying the organic layer and
`23
`solvent removal
`to give the crude product
`(1g).
`24 Solution of the crude solid in ethyl acetate then
`25 precipitation with ether résulted in white crystals of
`26
`the isobutylchloroformate derivative,
`,
`27
`.
`.
`28
`[4-(N-Hydroxyamino) -2R-isobutyl-3S-(4-(isobutyloxy-
`©)
`29
`carbonylamino) phenyl ) thiomethy1-succinyl ]-L-phenyl-_
`30 alanine-N~methylamide
`31
`_
`from example 26b was converted to the
`32 The product
`33
`hydroxamic acid as described in example 1g.
`to give a
`compound with the following characteristics.
`
`6 7 8
`
`AQUESTIVE EXHIBIT 1004 page 0715
`AQUESTIVE EXHIBIT 1004
`page 0715
`
`

`

`WO 90/05719
`
`PCT/GB89/01399
`
`65
`
`1 m.p. 198 - 200°C
`
`[alpha]) = -8.5° (c=1, methanol)
`
`2 3
`
`.
`
`4 5
`
`Analysis calculated for CayHyoN 405
`6 Requires: C61.41 H7.22 N9.55
`7
`Found:
`C62.04 H7.32 N9.67
`
`8 g
`
`(250MHz, Dg-DMSO) 9.60 (1H, s, NHOH), 8.83 (1H,
`delta,
`8S, NHOH), 8.31 (1H, 4, J = 8.5 Hz, CONH), 7.85 (1H, m,
`10
`CONHMe), 7.36 - 7.25 (4H, m, aromatic H), 7.14 - 7.05
`11
`(3H, m, aromatic H), 6.91 (2H, d, J = 8.5Hz, aromatic
`42
`13 H), 4.56 (1H, m, CHCH5Ph), 3.87 (2H, d,
`J = THz,
`44 OCH,CH(CH3)5), 2.92 (1H, dd, J = 13.7,4.0 Hz, CHCHjPh),
`15
`2.76 (1H, dd, J = 13.6,10 Hz, CHCH5Ph), 2.58 (3H, d, J
`16
`= 4-5 Hz, NHCH3), 2.50 - 2.34 (2H, m), 2.16 - 1.87 (3H,
`17. ™),
`1-35 (2H, m, CH9CH(CH3)2 and CH5CH(CH3)5),
`0.93
`ja
`(6H,
`Gd,
`J
`= 6.6Hz,
`OCH ZCH(CH3)9),
`0.87
`(1H,m,
`19
`CH5CH(CH3)5
`), 0.75 (3H,
`4d,
`J = 6Hz, CH(CH3)5), and
`20
`0-71 (3H, d, J = 6Hz, CH(CH3)5)-
`21
`
`22
`23 Example26
`
`24
`25
`26
`27
`28
`
`29
`
`30
`
`31
`
`32
`
`33
`
`[4-(N-Hydroxyamino)-2R-isobuty1-3S-(4-(N-methyl-N-
`(tertbutoxycarbonyl)-glycylamino) phenyl)thiomethyl-
`succinyl J-Lphenylalanine-N-methylamide.
`
`BOC.
`
`AQUESTIVE EXHIBIT 1004
`
`AQUESTIVE EXHIBIT 1004 page 0716
`
`page 0716
`
`

`

`WO90/05719
`
`.
`
`PCT/GB89/01399
`
`66
`
`ontnawbBWDNY
`
`WWWwwrndd)nSENYNNYNYNYNYHBBABABABeSBwBeaWNOoWOwOsIHDUUFWY|COWOMOYHUBwNYBAO'W
`
`[4-Hydroxy-2R-isobuty1-3S-(4-(N-methyl-N-(tert-
`a)
`butoxycarbonyl)gqlycylamino) phenyl)thiomethyl-
`succinyl ]-L-phenylalanine-N-methylamide was prepared as
`described in example 26b by substitution of N-BOC
`sarcosine for the acid component.
`
`deltay (250MHz, Dg-DMSO) 9.97 (1H, s, CO>H), 8.36 (1H,
`d, J = 8.5 Hz, CONH), 7.91 (1H, m, CONHMe), 7.48 (2H,
`/d, J = 8.5Hz, aromatic H), 7.40 - 7.05 (5H, m, aromatic
`H), 6.97 (2H,
`qd, J = 8.5Hz, aromatic H), 4.58 (1H,
`nm,
`CHCHjPh), 3.95 (2H, d, J = 9Hz, NCH5CO), 2.92 (4H, mid,
`CHCH2Ph and BOCNCH;), 2.76 (1H, dd, J = 13,10 Hz,
`CHCH)Ph), 2.58 (3H, d, J = 4.5 Hz, NHCH3), 2.50 - 2.09
`(4H, mj,
`1.46 - 1.33
`(11H,
`m + 2xs,
`(CH3)3C,
`CHoCH(CH3)-5 and CHZCH(CH3)o),
`0.87 (1H,
`Mm,
`CH2CH(CH3)2 ), 0.75 (3H, d, J = 6Hz, CH(CH3)5), and
`0.71 (3H, d, J = 6Hz, CH(CH3)5).
`
`[4—-(N-Hydroxyamino) -2R-isobutyl-3S-(4-(N-methyl- N-
`b)
`(tertbutoxycarbonyl )-glycylamino) phenyl) - thiomethyl-
`succinyl ]-Lphenylalanine-N-methylamide was prepared
`from the material produced in example 27a as described
`in example 1g.
`
`8.83
`s, CONHOH),
`deltay (250MHz, Dg-DMSO) 9.97 (1H,
`(1H, s, NHOH), 8.32 (1H, d, J = 8.5 Hz, CONH), 7.86
`(1H, m, CONHMe), 7.46 (2H,
`a4, J = 8.5Hz, aromatic H),
`7.28 - 7.00 (5H, m, aromatic H), 6.97 (2H, d, J=
`8.5Hz, aromatic H), 4.56 (1H, m, CHCH»Ph), 3.94 (2H, a,
`J = 9Hz, NCH5CO), 2.87 (4H, m+d, CHCH>Ph and BOCNCH3),
`2.76 (1H, m, CHCH7Ph), 2.57 (3H, d, J = 4.5 Hz, NHCH3),
`2.25 - 1.91 (2H, m), 1.42 - 1.30 (11H, m + 2xs,
`(CH3)3C,
`CH>CH(CH3) and CHCH(CH3)5), 0.92 (1H,
`nm,
`CHjCH(CH3)> ), 0.80 (3H, d, J = 6Hz, CH(CH3)5),
`and
`0.73 (3H, d, J=6Hz, CH(CH3)5).
`
`AQUESTIVE EXHIBIT 1004=page 0717
`
`AQUESTIVE EXHIBIT 1004 page 0717
`
`

`

`WO 90/05719
`
`PCT/GB89/01399
`
`67
`
`Example 27
`
`Collagenase inhibition activity
`
`of
`The potency
`as inhibitors
`
`compounds of general formula I to act
`of
`collagenase (a metalloproteas
`
`involved in tissue degradation) was determined by the
`
`1 2
`
`3 4
`
`5 6
`
`7
`
`8
`
`(Anal. Biochem., 99,
`procedure of Cawston and Barrett,
`9
`340-345, 1979), hereby incorporated by reference,
`10
`11. whereby a ImM solution of the inhibitor being tested or
`12 dilutions thereof was
`incubated
`at 37° for 16 hours
`
`13 with collagen and collagenase (buffered with 25mM
`14 Hepes,
`pH 7.5 containing 5mM CaCly5,
`0.05% Brij
`35 and
`15
`0.02% NaN3).
`The collagen was acetylated 146
`collagen
`16
`prepared by the method of Cawston and Murphy
`(Methods
`
`17 in_Enzymology, 80, 711, 1981), hereby incorporated by
`18
`reference,
`The
`samples’ were centrifuged to sediment
`
`19
`
`undigested collagen and an aliquot of
`
`the radioactive
`
`a scintillation
`removed for assay on
`20 Supernatant
`The collagenase
`a measure
`21
`.counter as
`of hydrolysis.
`22 activity in the presence of
`1
`mM inhibitor,
`or
`a
`23 dilution thereof, was compared to activity in a control
`24
`devoid of
`inhibitor and the results reported below as
`25
`that
`inhibitor concentration effecting 50% inhibition
`26
`of the collagenase (ICsq)-
`27
`28
`29
`30
`31
`
`Compound of Example No.
`1
`2
`7
`
`ICso
`20 nM
`8 nM
`.3 nM
`(50% @
`1 mcM)
`
`32
`
`33
`
`AQUESTIVE EXHIBIT 1004 page 0718
`AQUESTIVE EXHIBIT 1004
`page 0718
`
`

`

`WO 90/05719
`
`PCT/GB89/01399
`
`68
`
`. Example 28°
`
`Stromelysininhibition activity
`
`The potency of compounds of general formula I to act as
`inhibitors of stromelysin was determined using the
`procedure of Cawston et al (Biochem. J., 195, 159-165
`1981), hereby incorporated by reference, whereby a 1mM
`solution of the inhibitor being tested or dilutions
`thereof was
`incubated at 37°C for
`16 hours with
`stromelysin and 14, acetylate casein (buffered with
`25mM Hepes, pH 7.5 containing 5mM CaCl5, 0.05% Brij 35
`and
`0.02% NaN3.
`The casein was
`14c acetylated
`according to the method described in Cawston et al
`_(Biochem. J., 195,.159-165, 1981), hereby incorporated
`by reference. The stromelysin activity in the presence
`of 1mM, or a dilution thereof, was composed to activity
`in a control devoid of
`inhibitor and the results
`reported below as that inhibitor concentration
`effecting 50% inhibition of the stromelysin (ICcg).
`
`Compound of Example No.
`1
`2
`
`— iCsg
`10 nM
`20 nM
`
`Examples of unit dosage compositions are as follows:
`
`onyno®WYNY+
`
`
`
`WnA=OUBIHWNADHONIHUNawNY|OCw
`
`AQUESTIVE EXHIBIT 1004
`
`AQUESTIVE EXHIBIT 1004 page 0719
`
`page 0719
`
`

`

`WO 90/05719
`
`PCT/GB89/01399
`
`69
`
`1 2 3 4
`
`Example29
`
`Capsules:
`
`5 6
`
`7
`Per 10,000
`
`
`8 ingredientsPerCapsule Capsules_
`
`9
`
`10
`
`11
`
`12
`13
`14
`15
`
`1.
`
`2.
`3.
`
`Active ingredient
`
`Cpd. of Form. I
`
`40.0 mg
`
`Lactose
`Magnesium
`stearate
`
`150.0 mg
`
`4.0 mg
`194.0 mg
`
`400 g
`
`1500 g
`
`40 g
`1940 g
`
`16
`17 Procedure for capsules:
`
`18
`
`19
`
`20
`
`Step 1.
`
`Blend ingredients No.
`
`1 and No.
`
`2 ina
`
`suitable blender.
`
`Step 2.
`
`21
`22
`23 Step 3.
`24
`25
`26 Step 4.
`27
`
`through a No. 30 mesh
`
`Pass blend from Step 1
`(0.59 mm) screen.
`Place screened blend from Step 2 ina
`3 and
`suitable blender with ingredient No.
`blend until the mixture is lubricated,
`Fill into No.
`1 hard gelatin capsule shells
`on a capsule machine.
`
`28
`
`29
`
`30
`
`31
`
`32
`
`33
`
`AQUESTIVE EXHIBIT 1004=page 0720
`
`AQUESTIVE EXHIBIT 1004 page 0720
`
`

`

`WO 90/05719
`
`PCT/GB89/01399
`
`70
`
`boornvnwo&WYDNBH
`
`WWwWwWnNNNNNNNNNNPPPPEBPBPPppeBpWNROWDADAUFwWNHPHOHDAAUMBFWNHHO
`
`Example30
`
`Tablets:
`
`Ingredients
`
`Per Tablet
`
`Per 10,000
`Tablets
`
`1.
`
`Active ingredient
`Cpd. of Form. I
`Corn Starch
`2.
`3. Alginic acid
`4.
`Sodium alginate
`5. Magnesium
`stearate
`7
`
`40.0 mg
`. 20.0 mg
`. 20.0 mg
`20.0 mg
`
`1.3 mg
`101.3 mg
`
`.
`
`400 g
`200 g
`200 g
`200 g
`
`13 q
`1013 g
`
`.
`
`-
`
`Procedure for tablets:
`
`Step 1.
`.
`Step 2.
`
`Step 3.
`
`Step 4.
`
`Step 5.
`
`Step6.
`
`Blend ingredients No. 1, No. 2, No. 3 and No.
`4 in a suitable mixer/blender.
`Add sufficient water portionwise to the blend
`_ from Step 1 with careful mixing after each
`addition. Such additions of water and mixing
`until the mass is of a consistency. to permit
`its conversion to wet granules.
`The wet mass is. converted to granules by
`- passing it through an oscillating granulator
`: usingaNo. 8 mesh (2.38) Screen.
`|
`The wet granules are then dried in an oven at
`(140°F (60°C) until dry.
`The dry granules are lubricated with
`ingredient No. 5.
`The lubricated granules are compressed on a
`suitable tablet press.
`
`AQUESTIVE EXHIBIT 1004
`
`AQUESTIVE EXHIBIT 1004 page 0721
`
`page 0721
`
`

`

`WO 90/05719
`
`PCT/GB89/01399
`
`oOornnmkhWwNY
`
`
`
`
`
`WWWWNNDN)DNNNNNNNPPPPPPBRPPOnPoveadTaAUbhODSAaTAARBDNHPOUNSER
`
`71
`
`Example 31
`
`Intramuscular Injection:
`Ingredient
`Compound of Formula I
`Active ingredient
`
`1.
`
`2.
`
`Istonic buffer
`
`Permil.
`
`Per liter
`
`10.0 mg
`
`10 g
`
`q.s.
`
`solution pH 4.0.
`
`q.s.
`
`Procedure:
`Step 1.
`
`Dissolve the active ingredient in the buffer
`solution.
`Step 2. Aseptically filter the solution from Step 1.
`Step 3.
`The sterile solution is now aseptically
`filled into sterile ampoules.
`The ampoules are sealed under aspetic
`conditions.
`
`Step 4.
`
`Example 32
`
`Suppositories:
`
`1.
`
`2.
`
`3.
`
`Ingredients
`Compound of Form.
`Active ingredient
`Polyethylene Glycol
`1000
`
`I
`
`Polyethylene Glycol
`4000
`
`Per
`
`Per Supp.
`
`1,000 Supp
`
`40.0 mg
`
`40 g
`
`1350.0 mg
`
`1,350 g
`
`450.0 mq
`
`1840.0 mg
`
`450 ¢g
`
`1,840 g
`
`AQUESTIVE EXHIBIT 1004
`
`AQUESTIVE EXHIBIT 1004 page 0722
`
`page 0722
`
`

`

`WO 90/05719
`
`PCT/GB89/01399
`
`72
`
`owonnanuFWN
`
`wwWWwWNHNNNNNNNNNPPPPBBBPbpopWNnHFEOUAT
`RARUFWNPOWANTAUHRWNHO
`
`2 and No.
`
`3 together and
`
`Procedure:
`Step 1.
`“Melt ingredient No.
`stir until uniform.
`Dissolve ingredient No. 1 in the molten mass
`from Step 1 and stir until uniform.
`- Pour the molten mass from Step 2 into
`“suppository moulds and chill.
`Remove the suppositories from moulds and
`wrap.
`
`Step 2.
`_
`Step3.
`
`Step 4.
`
`Example 33 —
`
`Eye Ointment
`
`An appropriate amount of a compound of general formula
`I is formulated into an eye
`ointment base having the
`following composition:
`
`Liquid paraffin
`_ Wool fat
`Yellow soft paraffin
`
`Example 34
`
`Topical skin ointment.
`
`An appropriate amount of a compound of general formula
`I
`is formulated into a topical skin ointment base
`having the following composition:
`
`Emulsifying wax
`White soft paraffin
`Liquid paraffin.
`
`oP
`
`ao
`
`(at
`
`*
`
`AQUESTIVE EXHIBIT 1004=page 0723
`
`AQUESTIVE EXHIBIT 1004 page 0723
`
`

`

`WO 90/05719
`
`PCT/GB89/01399
`
`73
`
`wowwonanawmF&FWNPB
`
`WWWWNNYNYBHNYYHNYNNNYFPFPRPPFRPRPFPFPPFWNFPoOUOWANHDYWFfWHYFPOYWONHOOTFWNYKFOO
`
`CLAIMS
`
`1.
`
`A compound of general formula I:
`
`Oo
`
`RA
`
`ri
`R'SO,
`
`wherein:
`
`RS
`RY
`N—, ps
`N
`CONHOH
`
`H
`
`O
`
`(I)
`
`RI
`
`thiophenyl,
`phenyl,
`represents a C,-C,g alkyl,
`substituted phenyl, phenyl(C,-C,)alkyl,
`heterocyclyl,
`(Cy-C,) alkylcarbonyl or phenacyl or
`substituted phenacyl group; or when n = QO, Rt
`represents SR*, wherein R* represents a group:
`
`A
`|
`
`CONHOH
`
`R?
`
`R?
`
`represents a hydrogen atom or a C,-C¢ alkyl, C,-C¢
`alkenyl, phenyl(Cy-Cg)alkyl,
`cycloalkyl (C,-C,) alkyl or cycloalkenyl (C,-C,) alkyl
`group;
`
`represents an amino acid side chain or a C4-C¢
`alkyl,
`benzyl,
`(C,-Cg alkoxy)benzyl or
`benzyloxy (C,-C¢ alkyl) or benzyloxy benzyl group;
`
`AQUESTIVE EXHIBIT 1004 page 0724
`AQUESTIVE EXHIBIT 1004 page 0724
`
`

`

`WO 90/05719
`
`PCT/GB89/01399
`
`74
`
`wowamnannauFFWNPH
`
`10
`
`12
`12
`
`represents a hydrogen atom or a C,-C¢g alkyl group;
`
`R?
`
`represents a hydrogen atom or a methyl group;
`
`3
`
`A
`
`is an integer having the value 0, 1 or 2; and
`
`represents a C,-Cg hydrocarbon chain, optionaly
`substituted with one or more C1-C6 alkyl, phenyl
`or substituted phenyl groups;
`
`or a salt thereof.
`
`in which the
`2. A compound as claimed in Claim 1,
`chiral centre adjacent
`the substituent R> has
`S
`stereochemistry.
`
`A compound as claimed in Claim 1 or 2, wherein the
`3.
`chiral centre adjacent
`the substituent R? has R
`stereochemistry.
`.
`
`13
`14
`15
`16
`17
`18
`19
`20
`in which
`2 or 3,
`A compound as claimed in Claim 1,
`4.
`21
`Rt represents a hydrogen atom or a C,-C, alkyl, phenyl,
`22
`thiophenyl, benzyl, acetyl or phenacyl group.
`23
`.
`24
`A compoundas claimed in any one of Claims 1 to 4,
`5.
`25
`26 wherein R* represents a C,-C, alkyl group.
`270
`a |
`28
`6.
`A compound as claimed in any one of Claims 1 to 5,
`29 wherein R?
`represents
`a benzyl or
`30
`4—(C,-C,) alkoxyphenylmethyl or benzyloxybenzyl group.
`31
`OO
`|
`.
`32
`A compound as claimed in any one of Claims 1 to 6,
`7.
`33
`wherein R4 represents a C,-C, alkyl group.
`
`.
`
`*
`
`AQUESTIVE EXHIBIT 1004 page 0725
`AQUESTIVE EXHIBIT 1004
`page 0725
`
`

`

`WO 90/05719
`
`PCT/GB89/01399
`
`75
`
`
`
`wowonanonrebkWN
`
`WWwWwWNNNNNNNNNNPPPFPPPPBPFPRYONPOWODNAUBONPOWANAMB®wWHNhb5
`
`A compound as claimed in any one of Claims 1 to 7,
`g.
`wherein R° represents a hydrogen aton.
`
`[4-(N-Hydroxyamino) ~2R-isobuty1l-3S-(phenylthio-
`9.
`methyl) -succinyl ]-L-phenylalanine-N-methylamide,
`
`[4-(N-Hydroxyamino) -2R-isobutyl-3S-(thiophenylthio-
`methyl) succinyl ]-L-phenylalanine-N-methylamide,
`
`(4- (N-Hydroxyamino) -2R-isobuty1-3S-(benzylthiomethyl1)
`succinyl ]-L-phenylalanine-N-methylamide,
`
`[4-(N-Hydroxyamino) -2R-isobuty1-3S-(acetylthiomethy1)
`succinyl ]-L-phenylalanine-N-methylamide or
`
`[4-(N-Hydroxyamino) -2R-isobuty1-38-(thiolmethyl)
`succinyl ]~L-phenylalanine-N-methylamide
`
`[4-(N-Hydroxyamino) ~2R-isobuty1-3S-(pivaloylthiomethy1)
`succinyl ]-L-phenylalanine-N-methylamide
`
`(4-(N-Hydroxyamino) -2R~isobuty1-3S-~-(phenylthiomethyl)
`succinyl ]-L-phenylalanine-N-methylamide sodium salt
`
`[4-(N-Hydroxyamino) -2R-isobuty1-3S~ (4-methoxyphenyl-
`thiomethyl) succinyl ]-L-phenylalanine-N-methylamide
`
`(4-(N-Hydroxyamino) ~2R-isobuty1-3S~ (4-hydroxypheny1-
`thiomethyl) succinyl ]-L-phenylalanine-N-methylamide
`
`[4-(N-Hydroxyamino) -2R-isobuty1~-38~(2-thiophenethio-
`methyl) succinyl]-L-phenylalanine-N-methylamide sodium
`salt
`
`AQUESTIVE EXHIBIT 1004=page 0726
`
`AQUESTIVE EXHIBIT 1004 page 0726
`
`

`

`WO 90/05719
`
`:
`
`PCT/GB89/01399
`
`76
`
`[4-(N-Hydroxyamino) -2R-isobuty1-3S8- (4-methoxyphenyl-
`thiomethyl) succinyl]-L-phenylalanine-N-methylamide
`sodium salt
`-
`
`[4-(N-Hydroxyamino) -2R-isobutyl-3S-(4-tertbutylphenyl-
`thiomethyl) succinyl ]-L-phenylalanine-N-methylamide
`
`[4-(N~Hydroxyamino) -2R-isobuty1-3S- (2, 4-dimethylphenyl-
`thiomethyl) succinyl]-L-phenylalanine-N-methylamide
`
`.
`
`bis-S,S'~{ [4 (N-Hydroxyamino-2R-isobuty1-3S-(thiomethy1)
`succinyl ]-L-phenylalanine-N-methylamide} disulphide
`
`' [4- (N-Hydroxyamino)—2R-isobuty1-35S-(3-bromophenylthio- .
`methyl) succinyl]—-L-phenylalanine-N-methylamide
`
`[4-(N-Hydroxyamino) -2R-isobuty1-3S-(3-chlorophenylthio-
`methyl) succiny1]-L-phenylalanine-N-methylamide
`
`_ [4 (N-Hydroxyamino) -2R-isobuty1-3S- (3-methylphenylthio-
`methyl) succinyl ]-L-phenylalanine-N-methylamide
`
`[4-(N-Hydroxyamino) -2R-isobuty1-3S- (4-(N-acetyl) -amino-
`phenylthiomethyl) succinyl ]—-L-phenylalanine-N-methyl-
`amide
`
`[4-(N-Hydroxyamino) -2R-isobutyl-3S-phenylsulphinyl-
`_ methylsuccinyl]-L-phenylaianine-N-methylamide
`
`[4-(N-Hydroxyamino) -2R-isobuty1-3S-phenylsulphonyl-
`methylsuccinyl ]-L-phenylalanine-N-methylamide
`
`wordnunFwnPp
`
`WwWwWNNnNnNNNNHNNNNPPPPPepPpPpOnPow}
`OaDTAUekeONPOHANDURWHEOC
`
`AQUESTIVE EXHIBIT 1004 page 0727
`AQUESTIVE EXHIBIT 1004
`page 0727
`
`

`

`oe
`
`WO 90/05719
`
`PCT/GB89/01399
`
`77
`
`[4-(N-Hydroxyamino) -~2R-isobuty1-3S-thiophenylsulphinyl-
`methyl-succinyl]-L-phenylalanine-N-methylamide
`
`[4-(N-Hydroxyamino) -2R~isobuty1-3S-thiophenylsulphonyl-
`methyl-succinyl ]-L-phenylalanine-N-methylamide
`
`[4-(N-Hydroxyamino) -2R-isobuty1-3S-phenylsulphonyl-
`methyl-succinyl]-L-phenylalanine-N-methylamide sodium
`salt
`,
`
`(4-(N-Hydroxyamino) -2R-isobuty1-3S-(4-(isobutyloxy-
`carbonylamino) phenyl) thiomethyl-succinyl]-L-phenyl-
`alanine-N-methylamide
`
`[4-(N-Hydroxyamino) -2R-isobuty1-

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