CFAD v. Anacor, IPR2015-01776
`ANACOR EX. 2008 - 1/9
`
`

`
`Journal of Medicinal Chemistry
`
`EDITOR
`PHILIPS. PORTOGHESE
`Department of Medicinal Chemistry, College of Pharmacy, University of Minnesota, Minneapolis, Minnesota 55455
`
`Patrick E. Hanna
`
`Richard F. Borch
`Frank H. Clarke
`William T. Comer
`Edward J. Cragoe, Jr.
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`Journal of Medicinal Chemistry (ISSN
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`ANACOR EX. 2008 - 2/9
`
`

`
`J. Med. Chern. 1984, 27, 947-953
`
`947
`
`route.4 No biological activity has been reported on this
`compound (4).
`·
`Compounds 3a and 3b seem to elicit their antidepressant
`activity by inhibiting the reuptake of norepinephrine into
`terminal neuronal granules as the tricyclic antidepressants,
`amitriptyline and imipramine, do. This was demonstrated
`by the test involving 3H-NE uptake by the rat heart5
`widely employed in the evaluation of antidepressants in
`which 3a was more, and 3b less, active than imipramine.
`At the same time it was established that they do not cause
`release of norepinephrine. Also, in the amine pressor re(cid:173)
`sponse study in the dogs, 1 another test used to characterize
`antidepressants, 3a was more potent than amitriptyline
`and imipramine in potentiating the effect of nor-
`
`(4) Skvarchenko, V. R.; Koshkina, ~· P. Zh. Org. Khim. 1979, 15,
`2367.
`(5) Hertting, G.; Axelrod, J.; Whitby, L. G. J. Pharmacal. Exp.
`Ther. 1961, 134, 146.
`
`epinephrine and about as potent as the latter in anta(cid:173)
`gonizing the effect of phenethylamine.
`Compound 3a thus seems to be a potential potent an(cid:173)
`tidepressant possessing an unusual structure.
`Acknowledgment. I thank Drs. M. Cohen, D. H.
`Smith, J. M. Stump, J. Jainchill, R. Clark, and C. Smith
`for the biological data.
`
`Registry No. 1, 52003-32-4; 2, 90269-35-5; 3a, 90269-36-6;
`3a·HC1, 90269-37-7; 3b, 90269-38-8; 3b·HC1, 90269-39-9; anthra(cid:173)
`cene, 120-12-7.
`
`P. Rajagopalan
`Medicinal Chemistry Section
`Biomedical Products Department
`E. I. duPont de Nemours and Co.
`Wilmington, Delaware 19898
`Received February 13, 1984
`
`Articles
`
`Preparation and Antibacterial Activities of New 1,2,3-Diazaborine Derivatives and
`Analogues
`
`Maximilian A. Grassberger,* Friederike Turnowsky, and Johannes Hildebrandt
`
`Sandoz Forschungsinstitut Ges.m.b.H., Brunnerstrasse 59, A-1235 Wien, Austria. Received November 29, 1983
`
`1,2-Dihydro-1-hydroxy-2-(organosulfonyl)areno[d][1,2,3]diazaborines 2 (arene =benzene, naphthalene, thiophene,
`furan, pyrrole) were synthesized by reaction of (organosulfonyl)hydrazones of arene aldehydes or ketones with
`tribromoborane in the presence of ferric chloride., The activities of 2 against bacteria in vitro and in vivo (Escherichia
`coli) were determined and structure-activity relationships are discussed. Included in this study are 2,3-dihydro-
`1-hydroxy-2-{p-tolylsulfonyl)-1H-2,1-benzazaborole (3) and 1-hydroxy-1,2,3,4-tetrahydro-2-(p-tolylsulfonyl)-2,1-
`benzazaborine (4) as well as the carbacyclic benzodiazaborine analogue 4-hydroxy-3-{p-tolylsulfonyl)isoquinoline
`(7). The nature of the active species is briefly discussed.
`
`The antibacterial activities of 1,2-dihydro-1-hydroxy-
`2-(organosulfonyl)benzo-, furo-, and -thieno[d][1,2,3]dia(cid:173)
`zaborines are well-documented in the literature.1- 12 In the
`
`Scheme I
`
`H. Poszich, 6th In-
`(1) H. Mueckter, H. Huemer, H. Sous,
`ternational Congress of Chemotherapy, Vienna, June 1967,
`Proceedings I/2, 805.
`(2) H. Huemer, S. Herrling, and H. Mueckter, Ger. Offen.
`1670 346, 1970.
`(3) H. Huemer, H. Herrling, and H. Mueckter, Ger. Offen.
`1670 494, 1971.
`(4) S. Gronowitz, T. Dahlgren, J. Namtvedt, C. Roos, B. Sjoberg,
`and U. Forsgren, Acta Pharm. Suec., 8, 377 (1971).
`(5) S. Gronowitz, T. Dahlgren, J. Namtvedt, C. Roos, G. Rosen,
`B. Sjoeberg, and U. Forsgren, Acta Pharm. Suec., 8, 623
`(1971).
`(6) G. M. Davies, Brit. Pat. 1367163, 1974.
`(7) S. Herrling, 15th Interscience Conference on Antimicrobial
`Agents and Chemotherapy, Washington, DC, Sept 1975.
`(8) H. Mueckter, H. Sous, G. Poszich, and F. Lagler, 15th Inter(cid:173)
`science Conference on Antimicrobial Agents and Chemother(cid:173)
`apy, Washington, DC, Sept 1975.
`(9) D. Florentin, B. P. Roques, J. M. Metzger, and J.P. Colin,
`Bull. Soc. Chim. Fr., 2620 (1974).
`(10) H. M. A. van Wersch, S. Herrling, and H. Mueckter, Ger.
`Offen. 2 533 918, 1977.
`(11) D. Forbes and G. M. Davies, lOth International Congress of
`Chemotherapy, Zurich, Sept 1977.
`
`A= 0
`
`2a
`
`0 s
`
`2b
`
`I[J
`
`0
`
`2d
`
`0 N
`
`R'
`2e
`
`2c
`
`2,3,1-benzodiazaborine series the known structural varia(cid:173)
`tion is almost exclusively restricted to the organosulfonyl
`side chain. Apart from compounds unsubstituted on the
`benzene ring, 1- 3•10•13 only a few 5-substituted derivatives
`
`(12) G. Hoegenauer and M. Woisetschlaeger, Nature (London) 293,
`662 (1981).
`(13) B. W. Mueller, Helv. Chim. Acta, 61, 325 (1978).
`
`0022-2623/84/1827-0947$01.50/0 © 1984 American Chemical Society
`
`ANACOR EX. 2008 - 3/9
`
`

`
`948 Journal of Medicinal Chemistry, 1984, Vol. 27, No.8
`
`Grassberger, Turnowsky, Hildebrandt
`
`Table I. 2-( Organosulfonyl)-1,2-dihydro-1-hydroxy-2,3, 1-benzodiazaborines 2a
`
`7«R IH
`
`. s,
`?'. I
`7S02R2
`/N
`
`::::-..,_
`
`R
`
`6
`R
`
`R5
`
`R4
`
`2a
`H
`H
`H
`H
`H
`4-CH3C6H4
`1
`H
`H
`2
`H
`H
`CH3
`4-CH3C6H 4
`H
`H
`H
`3
`H
`CH3
`4-CH3C6H4
`H
`H
`H
`H
`CH3
`4-CH3C6H4
`4
`H
`H
`H
`5
`CH3
`CH3
`4-CH3C6H4
`H
`F
`H
`H
`H
`4-CH3C6H 4
`6
`H
`H
`H
`F
`H
`4-CH3C6H4
`7
`H
`H
`F
`H
`H
`8
`4-CH3C6H4
`H
`H
`H
`H
`Cl
`4-CH3C6H4
`9
`H
`H
`H
`H
`Cl
`4-CH3C6H4
`10
`H
`H
`H
`H
`Cl
`11
`4-CH3C6H4
`H
`H
`H
`Cl
`Cl
`12
`4-CH3C6H4
`H
`H
`H
`Cl
`Cl
`13
`4-CH3C6H4
`Br
`H
`H
`H
`H
`14
`4-CH3CsH4
`Br
`H
`H
`H
`H
`4-CH3C6H4
`15
`Br
`H
`H
`H
`H
`16
`4-CH3C6H4
`H
`H
`H
`H
`OH
`17
`4-CH3C6H4
`H
`H
`H
`H
`NH2
`18
`4-CH3CsH4
`H
`H
`H
`H
`N(CH3h
`4-CH3C6H4
`19
`H
`H
`H
`H
`4-CH3C6H4
`20
`N(CHah
`H
`H
`H
`H
`NHCOCH3
`21
`4-CH3C6H4
`H
`H
`H
`22
`H
`NCH2CH2CH2CH2
`4-CH3C6H4
`H
`H
`H
`Cl
`4-CH3C6H 4
`N(CHa)2
`23
`H
`H
`H
`H
`COOH
`4-CH3C6H4
`24
`H
`H
`H
`H
`25
`4-CH3C6H4
`CsHs
`H
`H
`H
`H
`OH
`26
`CsHs
`H
`F
`H
`H
`H
`27
`2,4,6-(CH3)aCsH2
`H
`F
`H
`H
`H
`28
`2,4,5-Cl3C6H2
`Br
`H
`H
`H
`H
`29
`2,4,5-Cl3CsH2
`F
`H
`H
`H
`H
`4-H2NCsH4a
`30
`Br
`H
`H
`H
`H
`31
`4-H2NCsHl
`H
`H
`H
`H
`CH3
`32
`4-H2NCsH4a
`H
`H
`H
`33
`H
`CH3
`2-Cl-4-H2NC6H 3b
`H
`H
`H
`H
`CH3 Br
`2-Cl-4-CH3CONHC6Ha
`34
`H
`H
`H
`H
`2-Cl-4-CH3CONHC6H 3
`35
`F
`H
`H
`H
`H
`4-02NC6H4
`36
`Br
`H
`H
`H
`H
`4-02NC6H4
`37
`H
`H
`H
`H
`H
`38
`CH3
`H
`H
`H
`H
`CH3
`CH3
`39
`H
`H
`H
`H
`CH3
`n-C3H 7
`40
`H
`H
`H
`H
`Cl
`n-CaH7
`41
`H
`H
`H
`H
`H
`(CH3hN
`42
`H
`H
`H
`H
`CH3
`43
`(CHahN
`a From the nitro derivative by reduction with FejHOAc. b From the N-acetyl derivative by hydrolysis.
`
`%yield mp, °C
`162-4
`78
`28
`179-180
`145-6
`70
`15Q-2
`78
`161-3
`83
`173
`76
`168-9
`81
`168
`63
`182-4
`26
`15Q-1
`71
`173
`52
`185
`39
`225
`71
`198-201
`35
`162-4
`82
`180
`76
`158
`69
`188-190
`25
`173-6
`37
`182
`68
`198-205
`31
`186-190
`43
`19Q-3
`30
`259-261
`6
`209-210
`50
`207-210
`48
`162-5
`18
`225-230
`74
`26Q-4
`32
`216-7
`84
`21Q-5
`67
`185-8
`71
`215-7
`60
`25Q-5
`67
`235-240
`16
`201-3
`59
`214-6
`67
`124-6
`40
`127-8
`63
`109-112
`64
`109-113
`52
`125-6
`45
`137-140
`38
`
`are described, 2 without referring to their biological activ(cid:173)
`ities.
`One major goal of our work was therefore to synthesize
`2,3,1-benzodiazaborines with various substituents on the
`benzene ring and to evaluate their influence on the a,nti(cid:173)
`bacterial activities. For that purpose a new synthetic route
`to arenodiazaborines had to be developed.
`A second point of interest was the question whether the
`arenodiazaborines themselves are the active species or if
`hydrolytic cleavage at the BN bond to give the corre(cid:173)
`sponding (dihydroxyboryl)arenes is necessary for biological
`activities.
`Chemi$try. At the beginning of our study, the only
`method described in the literature for the preparation of
`2,3,1-benzodiazaborines was the reaction of o-formyl(cid:173)
`benzeneboronic acids with hydrazine or hydrazine deriv(cid:173)
`atives.14 Since substituted o-formylbenzeneboronic acids
`are difficult to obtain, an alternative approach that would
`
`be more suitable for our purpose was investigated. We and
`B. W. Mueller independently found that (organo(cid:173)
`sulfonyl)hydrazones of many aromatic and heteroaromatic
`aldehydes and ketones can easily be converted to the
`corresponding diazaborines with trihaloborane in a Frie(cid:173)
`del-Crafts type reaction (Scheme 1).13•15 Although the
`reaction could also be carried out without catalysts, the
`addition of Lewis acids like A1Cl3 was preferable. It led
`not only to substantially shorter reaction times but also
`to higher yields. In our hands FeCl3 in boiling 1,2-di(cid:173)
`chloroethane gave the best results. AlC13, ZnCl2, and SnC14
`could be used as well, whereas no effect was observed with
`TiC14.
`As can be seen from the Tables J..,..VJ, alkyl, halogen (F,
`Cl, Br), amino, alkylamino, and acylamino are tolerated
`as substituents R on the aromatic ring. With R = alkoxy
`
`(14) M. J. S. Dewar, Adv. Chem. Ser., 42, 227 (1964).
`
`ANACOR EX. 2008 - 4/9
`
`

`
`New 1,2,3-Diazaborine Derivatives and Analogues
`
`Journal of Medicinal Chemistry, 1984, Vol. 27, No.8 949
`
`Table II. 2-( Organosulfonyl)-1,2-dihyro-1-hydroxythieno-
`[3,2-d] [1,2,3]diazaborines 2b
`
`Table IV. 2-( Organosulfonyl)-1,2-dihydro-1-hydroxyfuro(cid:173)
`[3,2-d] [1,2,3]diazaborine 2d
`
`2d
`1
`2
`.3
`
`4-CH3C6H 4
`4-CH3C6H 4
`2,4,5-Cl3C6H 2
`
`CH3
`Br
`Br
`
`%yield
`66
`25
`84
`
`mp, °C
`169
`168-170
`164-5
`
`Table V. 2-( Organosulfonyl)-1 ,2-dihydro-1-hydroxypyrrolo(cid:173)
`[3,2-d] [1,2,3]diazaborines 2e
`
`OH
`I
`
`2
`~S02R
`N
`
`OCB,
`
`I
`R5
`
`7
`R
`
`OH
`I
`«NSOR
`I 2
`N
`
`2
`
`R6
`
`Rs
`R2
`R7 % yield mp, °C
`2b
`4-CH3C6H4
`1
`17Q-2
`Br
`H
`90
`4-CH3C6H4
`2
`Br
`H
`87
`153-5
`3 CsHs
`19Q-4
`Br
`H
`66
`4
`2-CH3C6H4
`H
`Br
`47
`162-4
`5
`2-CH3C6H4
`H
`CH3
`78
`171-4
`2-ClC6H4
`6
`H
`Br
`59
`194-8
`2-ClC6H4
`7
`H
`20Q-3
`CH3
`69
`8
`2-ClC6H4
`C2H5 H
`76
`173-5
`2-Cl-4-CH3C6H3
`9
`Br
`H
`68
`196-8
`CH3 ' H
`10
`2-Cl-4-CH3C6H3
`74
`203-4
`11
`4-CH3C6H4
`Cl
`H
`81
`178-180
`12
`2,4,6-(CH3)aC6H2
`Br
`H
`64
`183-4
`13
`4-CH3CONHC6H4
`Br
`H
`.....,217 dec
`60
`14
`2-Cl-4-CH3CONHC6H3 Br
`H
`.....,256 dec
`44
`15
`CH3
`H
`H
`39
`132-4
`16
`Br
`H
`7
`C2Hs
`95
`17
`n-C3H7
`H
`H
`58
`133-7
`18
`n-C3H7
`CH3
`H
`51
`85-6
`19
`(CH3hCHCH2
`Br
`H
`51
`113-5
`
`2e
`1
`2
`3
`
`CaH7
`4-CH3C6H 4
`4-CH3C6H 4
`
`%yield
`35
`45
`12
`
`mp, °C
`125
`155-8
`147-9
`
`Table III. 2-(0rganosulfonyl)-1,2-dihydro-1-hydroxythieno-
`[2,3-d] [1,2,3]diazaborines 2c
`
`Scheme II
`
`OH
`I
`B
`~NSO,R2
`R6
`I
`N
`
`R2
`Rs
`%yield
`2c
`mp, °C
`14Q-6
`1
`Br
`4-CH3C6H 4
`66
`36a
`2
`4-CH3C6H 4
`110
`C2Hs
`3
`Br
`2-ClC6H 4
`197-202
`79
`35a
`2-ClC6H 4
`4
`177-8
`C2Hs
`73a
`185-6
`CH3
`2-Cl-4-CH3C6H 3
`5
`40a
`6
`75-6
`CH3
`n-C3H7
`35a
`n-C3H7
`7
`60
`C2Hs
`75-6.
`67a
`CH3
`8
`(CHahCHCH2
`a From the corresponding 3-formylthiophene-2-boronic acid
`with (organosulfonyl)hydrazine.
`·
`
`the corresponding hydroxy derivatives were obtained as
`a consequence of concomitant ether cleavage (e.g., 2a-26).
`Electron-withdrawing substituents (R = CN or COOH)
`led to low yields in the cyclization step (e.g., 2a-,.24).
`Likewise no cyclization was observed with tosylhydrazones
`of aldehydes such as pyridine-2-carboxaldehyde, 1-
`methylimidazole-2-carboxaldehyde, or 3-methyliso(cid:173)
`thiazole-4-carboxaldehyde. With derivatives of
`"electron-rich" heterocycles, such as thiophene or furan,
`good yields of diazaborines were obtained.
`Generally, (arylsulfonyl)- and (alkylsulfonyl)hydrazones
`are equally good substrates for the cyclization reaction.
`Only the reaction with (alkylsulfonyl)hydrazones of
`thiophene-3-carboxaldehydes failed, probably due to de(cid:173)
`composition of the formed diazaborine under the reaction
`conditions. 2-(Alkylsulfonyl) -1,2-dihydrothieno [ 2,3-d](cid:173)
`[1,2,3]diazaborines (2c) were therefore prepared from 3-
`formylthiopheneboronic acids with (alkylsulfonyl)(cid:173)
`hydrazines as described in the literature. 5
`2,3-Dihydro-1-hydroxy-2-(p-tolylsulfonyl)-1H-2,1-benz(cid:173)
`azaborole (3) and 1-hydroxy-1,2,3,4-tetrahydro-2-(p(cid:173)
`tolylsulfonyl)-2,1-benzazaborine (4) are close analogues of
`
`(15) M. Grassberger, Ger. Offen. 2750878, 1978.
`
`1. Et3NBH3 , 130°,210°
`2. H20
`
`n = 1 1
`33 %
`n = 2 ~ 48 %
`
`the corresponding 2,3,1-benzodiazaborine derivative 2a-l.
`They were prepared from N-tosylbenzylamine and N-to(cid:173)
`syl-2-phenylethylamine, respectively, with triethylamine(cid:173)
`borane via pyrolytic ring closure19 (Scheme II).
`For biological comparison with the 2,3,1-benzo(cid:173)
`diazaborine 2a-l, the boron-free analogue 4-hydroxy-3-
`(p-tolylsulfonyl)isoquinoline (7) was prepared in four steps
`from phthalic acid anhydride (Scheme III).
`
`Biological Results and Discussion
`As observed earlier with other diazaborine deriva(cid:173)
`tives, 1•4•5•11 the antibacterial activity is almost exclusively
`confined to Gram-negative bacteria, including Neisseria
`gonorrhoea. This specificity has been explained on the
`basis of the mode of action of these derivatives which have
`·been shown to inhibit the biosynthesis of the lipopoly(cid:173)
`saccharide of Gram-negative bacteria.12
`Particularly good activity is shown against Proteus,
`Klebsiella, and Salmonella and a somewhat lower activity
`against Escherichia coli and Enterobacter (compare Tables
`
`(16) M. J. S. Dewar in "Progress in Boron Chemistry", H. Steinberg
`and A. L. McCloskey, Ed., Macmillan, New York, 1964, Vol.
`1.
`(17) M. J. Cook, A. R. Katritzky, and P. Linda, Adv. Heterocycl.
`Chem., 17, 255 (1974).
`(18) S. Gronowitz, J. Heterocycl. Chem., Suppl. 3, S-17 (1976).
`(19) Compare: R. Koester, K. Iwasaki, S. Hattori, andY. Morita,
`Justus Liebigs Ann. ()hem., 720, 23 (1968).
`
`ANACOR EX. 2008 - 5/9
`
`

`
`950 Journal of Medicinal Chemistry, 1984, Vol. 27, No.8
`
`Grassberger, Turnowsky, Hildebrandt
`
`Table VI. Antibacterial Activity of 2aa
`
`MIC values, J,LgjmL
`
`E. aerogenes
`~220
`>50
`
`K. pneumoniae
`~217
`3.12
`
`P. mirabilis
`~89
`12.5
`
`N. gonorrhoeae
`AS 7720 W2
`2
`8
`
`10
`1.56
`>50
`25
`6.25
`
`6.25
`12.5
`
`6925
`6.25
`
`50
`
`5
`3.12
`>50
`3.12
`6.25
`
`3.12
`25
`
`1.56
`12.5
`
`50
`
`2
`1
`
`2
`2
`
`1
`1
`
`1
`2
`
`8
`8
`
`8
`8
`
`2
`>8
`
`2
`8
`
`>50
`
`>50
`
`50
`50
`>50
`50
`10
`1.56
`1.56
`0.78
`1.25
`3.12
`
`0.78
`
`50
`10
`25
`6.25
`6.25
`0.78
`0.39
`0.39
`1.25
`1.56
`
`3.12
`
`>8
`
`>8
`
`1
`8
`1
`2
`1
`0.5
`1
`
`1
`
`8
`8
`2
`8
`1
`1
`2
`
`8
`
`>50
`50
`>50
`>50
`>50
`
`>50
`>50
`
`>50
`>50
`
`>50
`
`>50
`
`>50
`>50
`>50
`>50
`50
`25
`25
`10
`25
`50
`
`>50
`
`>300
`"'100
`>100
`>300
`>300
`65
`16
`5
`14
`16
`83
`"'100
`"'25
`
`42
`73
`14
`57
`
`S. typhimurium
`E. coli
`~120
`no. ED50, mgjkg
`~119
`6.25
`25
`1
`"'25
`>50
`2
`>50
`"'100
`50
`>50
`3
`>100
`3.12
`12.5
`4
`"'15
`5
`>50
`>50
`>600
`25
`>50
`6
`"'5
`6.25
`25
`7
`"'25
`8
`>50
`>50
`"'250
`>50
`>50
`9
`"'100
`3.12
`25
`10
`"'20
`12.5
`11
`>50
`"'50
`>50
`>50
`12
`>100
`13
`>50
`>50
`>100
`>50
`14
`>50
`>100
`6.25
`25
`15
`"'40
`6.25
`16
`50
`"'30
`17
`>50
`>50
`"'300
`>50
`>50
`18
`>300
`>50
`>50
`19
`>300
`>50
`20
`>50
`"'100
`>50
`>50
`21
`>300
`>300b
`>50
`>50
`22
`>50
`23
`>50
`>300
`24
`>50
`>50
`>50
`>50
`25
`12.5
`50
`26
`27
`>50
`>50
`>50
`>50
`28
`>50
`>50
`29
`25
`10
`30
`3.12
`10
`31
`3.12
`6.25
`32
`3.12
`10
`33
`6.25
`25
`34
`12.5
`50
`35
`50
`>50
`36
`3.12
`6.25
`37
`38
`>50
`>50
`12.5
`25
`25
`>50
`39
`6.25
`>8
`>8
`2.5
`10
`40
`6.25
`2
`2
`1.56
`1.25
`41
`2
`2
`2.5
`3.12
`25
`10
`1.56
`50
`42
`50
`12.5
`>50
`>50
`>8
`>8
`12.5
`43
`>50
`>50
`8
`8
`6.25
`6.25
`a The minimum inhibitory concentrations (MICs) were determined by serial broth dilutions in trypticase soy broth after incubation for 16
`h at 37 °C. The inocula were 10c 105 colony forming units. The MIC values for Neisseria gonorrhoeae were determined by the agar dilution
`test (Kellogg agar) with 105 colony forming units after incubation at 36 °C in 5% C02 for 20 h. The MIC is defined as the lowest concen-
`tration that inhibited visible growth. The ED50 values were determined in NMRI mice infected intraperitoneally with E. coli 120 and
`treated po immediately and 5 h after the infection. b Subcutaneous application.
`
`Scheme Ill
`
`~0 TosMIC a: S02 -Q-cH3
`
`OA::-N
`
`COOH
`
`0
`
`0
`
`Et3NBH/Et
`
`0BF
`2
`3
`
`ct:-o-
`
`so2
`CH20H
`
`_
`
`CH3
`
`5
`
`VI-IX). Pseudomonas aeruginosa was not found to be
`susceptible to diazaborine compounds.
`As can be seen from Table VI (entries 1-16), substitution
`
`by methyl or halogen (F, Cl, Br) on the benzene ring of
`2a (R2 = p-tolylsulfonyl, R4 =H) has no marked influence
`on the antibacterial activities in vitro. Generally, deriva-
`
`ANACOR EX. 2008 - 6/9
`
`

`
`New 1,2,3-Diazaborine Derivatives and Analogues
`
`Journal of .Medicinal Chemistry, 1984, Vol. 27, No. 8 951
`
`Table VII. Antibacterial Activity of 2ba
`
`E. coli
`no. EDs0, mgjkg
`~120
`1
`,....,15
`6.25
`>50
`2
`>300
`3
`,....,20
`3.12
`4
`25
`57
`25
`5
`50
`25
`39
`6
`12.5
`7
`49
`3.12
`49
`8
`50
`9
`39
`50
`10
`32
`25
`11
`28
`>50
`12
`>600
`55
`25
`13
`14
`50
`208
`>50
`15
`113
`12.5
`16
`6.25
`28
`17
`1.56
`18
`4.5
`12.5
`73
`19
`a See footnote a of Table VI.
`
`E. aerogenes
`~220
`25
`>50
`12.5
`50
`50
`25
`12.5
`12.5
`50
`50
`50
`>50
`25
`25
`>50
`12.5
`25
`3.12
`12.5
`
`Table VIII. Antibacterial Activity of 2ca
`
`E. coli
`no. ED50, mgjkg
`~120
`>50
`>300
`1
`6.25
`2
`9
`>50
`>300
`3
`10
`24
`4
`25
`50
`5
`1.25.
`2
`6
`2.5
`7
`4
`6.25
`5
`8
`a See footnote a of Table VI.
`
`E. aerogenes
`~220
`>50
`25
`>50
`25
`50
`1.56
`5
`6.25
`
`Table IX. Antibacterial Activity of 2d-ha
`
`E. coli
`no. ED50, mgjkg
`~120
`,....,10
`12.5
`2d-1
`2d-2
`12.5
`"'30
`2d-3
`>50
`>300
`2e-1
`>50
`>300
`2e-2
`>50
`>300
`2e-3
`>50
`>600
`>50
`>100
`2f
`2g
`>50
`>300
`2h
`>50
`>600
`a See footnote a of Table VI.
`
`E .. aerogenes
`~220
`25
`25
`>50
`>50
`nt
`>50
`>50
`>50
`>50
`
`MIC values, JLg/mL
`
`S. typhimurium
`~119
`3.12
`>50
`0.78
`6.25
`6.25
`3.12
`1.56
`0.78
`12.5
`12.5
`5
`>50
`3.12
`3.12
`>50
`3.12
`3.12
`0.78
`1.56
`
`K. pneumoniae
`~217
`1.56
`>50
`0.78
`3.12
`0.78
`1.56
`1.56
`0.31
`6.25
`1.56
`2.5
`25
`3.12
`3.12
`>50
`3.12
`1.56
`0.39
`1.56
`
`P. mirabilis
`~89
`1.56
`50
`1.56
`1.56
`1.56
`0.78
`1.56
`0.39
`3.12
`1.56
`1.56
`2.5
`3.12
`6.25
`>50
`6.25
`25
`0.78
`6.25
`
`N. gonorrhoeae
`AS 7720
`W 2
`0.5
`2
`
`1
`0.5
`0.5
`0.5
`0.5
`0.5
`0.25
`0.5
`1
`1
`1
`1
`>8
`2
`8
`1
`2
`
`2
`0.5
`0.5
`1
`0.5
`1
`1
`1
`1
`2
`1
`1
`>8
`2
`8
`1
`2
`
`MIC values, JLg/mL
`
`S. typhimurium
`~119
`>50
`1.56
`>50
`2.5
`6.25
`0.19
`0.31
`1.56
`
`K. pneumoniae
`~217
`>50
`1.25
`>50
`1.25
`3.12
`0.31
`0.31
`0.78
`
`P. mirabilis
`~89
`25
`1.25
`>50
`1.56
`1.56
`1.56
`1.25
`1.56
`
`N. gonorrhoeae
`AS 7720
`W 2
`>8
`>8
`0.25
`0.5
`>8
`>8
`0.5
`1
`1
`1
`0.5
`1
`0.5
`0.5
`1
`1
`
`MIC values, JLg/mL
`
`S. typhimurium
`~119
`3.12
`3.12
`50
`>50
`>50
`>50
`>50
`>50
`>50
`
`K. pneumoniae
`~217
`1.56
`3.12
`50
`25
`nt
`50
`25
`>50
`>50
`
`P. mirabilis
`~89
`3.12
`3.12
`6.25
`>50
`nt
`50
`3.12
`50
`50
`
`N. gonorrhoeae
`AS 7720 W 2
`1
`8
`2
`8
`0.5
`2
`>8
`>8
`nt
`nt
`nt
`nt
`2
`8
`>8
`>8
`2
`8
`
`tives with methyl or halogen in position 5 or 7 are less
`active than the unsubstituted parent compound 2a-l. The
`7-bromo derivative 2a-16 is the most active compound out
`of this group and shows the same activity as 2a-l. Sub(cid:173)
`stitution (methyl, halogen) in position 6 has practically no
`influence on the antibacterial activities.
`The in vivo data (E. coli septicaemia) follow the same
`trend. The only exception is the 5-fluoro derivative 2a-6,
`for which a surprisingly low ED50 value has been deter(cid:173)
`mined. We have no explanation so far for this phenome(cid:173)
`non. In metabolic studies20 its main metabolite was iso(cid:173)
`lated from the urine, which, however, exhibited only a
`
`(20) F. Battig, unpublished results.
`
`slightly better antibacterial activity as compared to the
`parent compound.
`Substitution of 2a with polar groups (OH, NH2, NR2,
`NHCOCH3, COOH) in position 6 or 7 (Table VI, entries
`17-24) generally leads to complete loss of activity. Only
`the 7-hydroxy derivative 2a-27 is slightly active in vitro.
`Regarding the organosulfonyl side chain, replacement
`of p-tolylsulfonyl by (4-aminophenyl)sulfonyl enhances the
`antibacterial activities (Table VI, entries 3Q-33). The
`5-fluoro derivative 2a-30 however, does not show the ex(cid:173)
`ceptional increase in the in vivo activity that was observed
`in the p-tolylsulfonyl series (2a-6). Against E. coli 120 the
`order of activity in vitro and in vivo is 5-F < 6-Br < 6-Me.
`Derivatives of 2a with alkylsulfonyl in position 2 and
`
`ANACOR EX. 2008 - 7/9
`
`

`
`952 Journal of Medicinal Chemistry, 1984, Vol. 27, No.8
`
`Grassberger, Turnowsky, Hildebrandt
`
`chlorine or methyl in position 6 (Tables VI, entries 39-41)
`showed surprisingly low MIC and ED50 values.
`In the thienodiazaborine series, the thieno[2,3-d]diaza(cid:173)
`borines 2b are generally slightly more active than their
`thieno[3,2-d] counterparts 2c (Tables VII and VIII) with
`the exception of the 6-bromo derivatives of 2c (Table VIII,
`entries 1, 2), which are inactive. Remarkably good activ(cid:173)
`ities in vitro and in vivo were observed with the 2-alkyl(cid:173)
`sulfonyl derivatives of 2b and 2c. Compound 2b-18 has
`been selected for further evaluation.
`As in the thienodiazaborines (2b), substitution of bro(cid:173)
`mine instead of methyl in position 6 of the furo(cid:173)
`diazaborines 2d also brings no advantage in biological
`activity (Table IX). With the oligocyclic diazaborine
`derivatives 2f-2h, only very low activities in vitro were
`observed (Table IX). The pyrrolo compounds 2e are to(cid:173)
`tally inactive (Table IX).
`
`OH
`I
`
`N-Tos
`I
`N
`
`+
`
`1:1 2h
`
`41 %, mp 191-3°C
`
`OH
`I
`
`N:n:-..:: 'N-Tos
`I
`I
`I
`Tos"'N'B ~ ""N 2g
`I
`"-'
`OH
`24 %, mp
`
`> 250°C
`
`Variation of the diazaborine ring itself led to negative
`results: the benzazaborole 3 is only slightly active against
`Klebsiella and Proteus in vitro. No antibacterial activity
`was found with the tetrahydrobenzazaborine 4.
`One major problem with the diazaborines is their in(cid:173)
`herent toxic potential, which is probably due to the
`
`Tos NHN"-<B (0H) 2
`ILJ s
`
`OH
`I
`~~-Tos
`\~N
`
`OH
`I
`
`N- Tos
`I
`N
`
`10
`
`11
`
`12
`
`r(YB(OHh
`~NN-Tos
`I
`CH 3
`
`13
`""'
`
`structural element of areneboronic acid amide. When
`designing boron-free analogues, a very important question
`has to be considered; either the bicyclic arenodiazaborines
`themselves are the active species or the BN bond has to
`be cleaved during or prior to the interaction of the mole(cid:173)
`cule with the biological target. Gronowitz et al.4 reported
`that the tosylhydrazone 8 of 4-(dihydroxyboryl)(cid:173)
`thiophene-3-carboxaldehyde, which does not readily cyclize
`to the corresponding diazaborine, has about the same weak
`
`activity against Gram-negative bacteria as the cyclic ana(cid:173)
`logues 9 and 10, although no detailed biological data were
`given for 8. Studies11 with the highly active furo(cid:173)
`diazaborine 11 showed that the product of hydrolytic
`cleavage at the BN bond 12, which is the main metabolite
`isolated from animals, has only slight antibacterial activ(cid:173)
`ities. However, it cannot be excluded that at least partial
`recyclization might occur under in vivo conditions.
`To avoid the problem of potential ring opening and
`recyclization, we decided to study the biological activities
`of analogues where such reactions are principally not
`possible. The isoquinoline 7 was selected as a stable
`analogue of benzodiazaborine 2a-l, since various stud(cid:173)
`ies14·16-17 have shown that replacement of a C-C unit in
`aromatic compounds by the isoelectronic BN group leads
`to heterocycles with similar chemical and physical prop(cid:173)
`erties. If 2a-l in its bicyclic form represents the active
`species, one should expect to find at least some biological
`activity with the carbacyclic analogue. However, we found
`that in all test systems 7 was completely inactive.
`TheN-methyl derivative 13 was prepared as a stable
`ring-opened analogue to 2a-l This compound inhibited
`the growth of the E. coli strain PL 2 but was at least 10
`times less active than the corresponding bicyclic diaza(cid:173)
`borine 2a-l (data not shown). Benzeneboronic acid had
`a much lower effect on the growth of the E. coli strain.
`Since neither of our analogues (7, 13) showed activities
`similar to 2a-l, no conclusion can be drawn from our ex(cid:173)
`periments, although they do support to some extent the
`hypothesis that ring opening is a necessary event for an(cid:173)
`tibacterial action. The lower activities of 13 as compared
`to 2a-l might be caused either by a reduced transport of
`the ring-opened compound to the target or by the addi(cid:173)
`tional methyl group in 13, causing a less favorable inter(cid:173)
`action between the compound and the target. More de(cid:173)
`tailed studies on the mode of action of these compounds
`are presently being carried out.
`
`Experimental Section
`
`1,2-Dihydro-5-fluoro-1-hydroxy-2-(p-tolylsulfonyl)-2,3,1-
`benzodiazaborine (2a-6). To 0.4 g (2.5 mmol) of FeC13 in 200
`mL of dry ClCH2CH2Cl were added simultaneously from two
`dropping funnels with intensive stirring under argon atmosphere
`10 mL (26 g, 104 mmol) of BBr3 in 30 mL of ClCH2CH2Cl and
`10 g (34 mmol) of o-fluorobenzaldehyde tosylhydrazone in 500
`mL of ClCH2CH2Cl within 5 min. The mixture was heated under
`reflux for 20 min, cooled to 5 °C, and poured into 300 mL of ice
`water. The organic phase was separated, washed with water (2
`X 50 mL), and extracted with 1 N NaOH (3 X 200 mL). After
`acidification of the water phase (pH 2-3), the product was
`reextracted with CH2Cl2• On evaporation of the dried (MgSO 4)
`CH2Cl2 solution, 8.3 g (76%) of 2a-6 was obtained: white crystals,
`mp 173 oc. Anal. (C14H 12BFN20 3S) C, H, N, S.
`2-[(4-Aminophenyl)sulfonyl]-6-bromo-1,2-dihydro-1-
`hydroxy-2,3,1-benzodiazaborine (2a-31). To 1.7 g (4.1 mmol)
`of 6-bromo-1 ,2-dihydro-1-hydroxy-2-[ ( 4-nitrophenyl)sulfonyl]-
`2,3,1-benzodiazaborine in 15 mL of acetic acid was added 0.94
`g of iron powder at 5o-60 °C with intensive stirring. The mixture
`was heated to 7Q-80 °C for 45 min. After addition of water and
`filtration, the crude crystalline product separated from the filtrate
`on cooling to room temperature. Recrystallization from DMF(cid:173)
`water gave 1.08 g (68% ): light yellow crystals, mp 21Q-215 °C.
`Anal. Calcd for C13H 11BBrN30 3S: C, 41.09; H, 2.92; N, 11.06.
`Found: C, 40.65; H, 3.00; N, 11.22.
`2-[ 4-Amino-2-chlorophenyl )s ulfony l]-1,2-dihydro-6-
`methyl-2,3,1-benzodiazaborine (2a-33). 2-[(4-Acetamido-2-
`chlorophenyl)sulfonyl]-1,2-dihydro-1-hydroxy-6-methyl-2,3,1-
`benzodiazaborine (2.6 g, 6.6 mmol) in 160 mL of THF and 20 mL
`of concentrated aqueous HCl were stirred for 7 days at room
`temperature. The solution was then concentrated in vacuo to a
`volume of 4