JMCMAR
`
`· VOLUME 18 :
`
`JANUA,RY 1975
`'
`
`' .
`
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`,·_,
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`! ~~,.
`
`·.(j
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`·.· ... Cf
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`NUMBER 1
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`I
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`¥J ,•' I
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`_.,,
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`. . . . . ··' ..
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`. .
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`Ex. 1091-0001
`
`

`

`Journal of Medicinal Chemistry
`
`EDITOR
`PHILIP S. PORTOGHESE
`Department of Medicinal Chemistry, College of Pharmacy, University of J,finnesota, Minneapolis, Minnesota 55455
`
`Mahmoud M. Abdel-Monem
`
`Joseph G. Cannon
`Ralph E. Christoffersen
`Paul N. Craig
`John W. Daly
`
`Louis S. Harris
`
`ASSOCIATE EDITORS
`Patrick E. Hanna
`John L. Neumeyer, Book Review Editor
`
`EDITORIAL ADVISORY BOARD
`Eugene C. Jorgensen
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`Hobert A. Maxwell
`
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`
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`K. L. Rinehart, Jr.
`Andre Rosowsky
`Robert A. Scherrer
`Monroe E. Wall
`
`EX-OFFICIO: Barbara E. Roth (Secretary, Division of Medicinal Chemistry)
`
`Published by the
`AMERICAN CHEMICAL SOCIETY
`1155 16th Street, N. W.
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`© Copyright, 1974, by the American
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`This material was Hipied
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`~ubject US Copyright Laws
`
`Ex. 1091-0002
`
`

`

`Journal of Medicinal Chemistry
`
`Volume 18, Number 1
`January 1975
`
`,JMCMAR lS(i) 1-122 (1975)
`ISSN 0022-2623
`
`Benzopyranopyridine Derivatives. 1. Aminoalkyl Derivatives of the Azaxanthenes as Bronchodilating Agents
`. . . Frank J. Villani,* Thomas A. Mann, Elizabeth A. Wefer,
`,Janet Hannon, Louis L. Larca,
`Mildred ,J. Landon, William Spivak, Dhiru Vashi, Salvatore Tozzi, Gisela Danko, Mauricio del Prado,
`· and Robert Lutz
`·
`
`Carbocyclic Prostaglandin Analogs. 1. Steroid Carboxylic Acids
`. . . D. L. Venton, R. E. Counsell,* J. H. Sanner, and K. Sierra
`
`Aromatic Amino Acid Hydroxylase Inhibitors. 4. 3-Substituted a-Methyltyrosines
`. . . Ahmed H. El Masry, Souheir E. El Masry, Larry E. Hare, and Raymond E. Counsell*
`
`Effect of Acylation with Eleostearic Acids on the Monoamine Oxidase Inhibitory Potency of Some Hydrazine
`Antidepressants in Mice . . . . . . . . . . . . . . . Song Y. Hsu,* Chian L. Huang, and Irving W. Waters
`
`Comparison of the Effects of Histamine and Tolazoline on Adenylate Cyclase Activity from Guinea Pig Heart
`. . . Ira Weinryb* and Inge M. Michel
`
`Synthesis and Microbiological Activities of Some Monohalogenated Analogs of Tyrosine
`. . . Tommy J. McCord,* David R. Smith, Douglas W. Winters, -John F. Grimes, Karen L. Hulme,
`Lawrence Q. Robinson, Larry D. Gage, and Alvie L. Davis
`
`The Charge-Transfer Constant. A New Substituent Constant for Structure-Activity Relationships
`. . . Bogumil Hetnarski* and R. D. O'Brien
`
`Studies on Antianaphylactic Agents. 4. Synthesis and Structure-Activity Relationships of
`3-( 4-Oxo-4H-1-benzopyran-3)acrylic Acids, a New Series of Antiallergic Substances, and Some Related
`Compounds . . . . . · Akira Nohara,* Hisashi Kuriki, Taketoshi Saijo, Kiyoshi Ukawa, Tadakazu Murata,
`Morio Kanno, and Y asushi Sanno
`
`1
`
`9
`
`16
`
`20
`
`23
`
`26
`
`29
`
`Synthesis and Absolute Stereochemistry of 5-Alkyl-5-(3'-hydroxy-1' -methylbutyl)barbituric Acid and
`5-Alkyl-5-(3' -hydroxy-1' -methylbutyl)-2-thiobarb\turic Acids . . . . . . . . F. I. Carroll* and G. N. Mitchell
`Aminobenzoic Acid Diuretics. 7. 3-Substituted 4-Phenyl-, 4-Arylcarbonyl-, and 4-Arylmethyl-5-sulfamoylbenzoic Acids
`and Related Compounds . .
`Ole B. Tvaermose Nielsen, Herta Bruun, Claus Bretting, and Peter W. Feit*
`John B. Hill,* Richard E. Ray, H. Wagner, and Richard L. Aspinall
`Antiinflammatory Sydnones. 2. . .
`2-Aryl-5-benzoxazolealkanoic Acid Derivatives with Notable Antiinflammatory Activity
`. . . David W. Dunwell, Delme Evans,* Terence A. Hicks, Colin II. Cashin,* and Ann Kitchen
`Antitumor Agents. 11. Synthesis and Cytotoxic Activity of Epoxides of Helenalin Related Derivatives
`. . . Kuo-Hsiung Lee,* Sun-Hyuk Kim, Hiroshi Furukawa, Claude Piantadosi, and Eng-Shang Huang
`
`Preparation and Anti tumor Activity of a Rearranged Ester of Cephalotaxine
`. . . Kenneth L. Mikolajczak,* Richard G. Powell, and Cecil R. Smith, .Jr.
`
`34
`
`:37
`
`41
`
`50
`
`53
`
`59
`
`63
`
`Preparation and Antileukemic Activity of Some Alkoxybenzo[c)phenanthridinium Salts and Corresponding
`Dihydro Derivatives . . . . . . . . . . . . . . . . . . . . . Robert IL-Y. Zee-Cheng and C. C. Cheng*
`Quinuclidine Chemistry. :J. (J-cis-2-( 4' -Chlorobenzhydryl)-3-quinuclidinol, a New Central Nervous System Stimulant.
`Importance of the Benzhydryl Configuration . . . . . . . E .. J. Warawa,* N. ,J. Mueller, and Jonas A. Gylys
`Pyrido[2,3-d]pyrimidine Antibacterial Agents. 3. 8-Alkyl- and
`8-Vinyl-5,8-di_hydro-5-oxo-2-( l-piperazinyl)pyrido[2,3-d]pyrimidine-6-carboxylic Acids and Their Derivatives
`. . . Jun-ichi Matsumoto* and Shinsaku Minami
`Purinylhydantoins. Facile Conversion of the Naturally Occurring N-(Purin-6-ylcarbamoyl)-L-amino Acids into
`3-Purin-6-ylhydantoins and 3-Cyclohexyl-1-(purin-6-ylcarbamoyl) hydantoins
`. . . Chung Il Hong and Girish B. Chheda*
`
`Quantitative Structure-Activity Relationships for Dicoumarol Antivitamins Kin the Uncoupling of Mitochondrial
`Oxidative Phosphorylation . . . . . . . Rosine Labbe-Bois, Claude Laruelle, and ,Jean-,Jacques Godfroid*
`
`Amidines and Related Compounds. 6. Studies on Structure-Activity Relationships of Antihypertensive and
`Antisecretory Agents Related to Clonidine
`Timothy Jen, Helene Van Hoeven, William Groves, Richard A. McLean, and Bernard Loev*
`
`66
`
`71
`
`74
`
`79
`
`85
`
`90
`
`This material was C'f~ed
`atthe NLM and mav bE
`
`Ex. 1091-0003
`
`

`

`2A Journal of Medicinal Chemistry, 1975, Vol. 18, No. 1
`
`NOTES
`
`.
`Synthesis and Biological Actions of Fragmented Derivatives ofTetrahydroisoquinolines
`. . . Duane D. Miller,* William V. P. Merritt, Peter F. Kador, and Denms R. Feller
`
`99
`
`/3-Cocaine . . . . . . . . . . . .
`
`. . . . . . . . . . . . . . . .
`
`. Robert L. Clarke* and Sol J. Daum 102
`
`Synthesis of Chlorozotocin, the 2-Chloroethyl Analog of the Anticancer Antibiotic Streptozotocin
`. . . Thomas P. Johnston,* George S. McCaleb, and John A. Montgomery 104
`
`Synthesis and Antimicrobial Evaluation of N-n-Alanyl-1-aminoethylphosphonic Acid
`. . . Joseph W. Huber, III, W. Franklin Gilmore,* and Larry W. Robertson 106
`
`Centrally Acting Emetics. 8. Conformational Aspects of Certain Dihydrophenanthrene Congeners of Apomorphine
`. . . Joseph G. Cannon,* Robert V. Smith, Mohd. A. Aleem, and John Paul Long 108
`
`Centrally Acting Emetics. 9. Hofmann and Emde Degradation Products of Nuciferine
`. . . ,Joseph G. Cannon,* Plaston R. Khonje, and John Paul Long
`
`llO
`
`Antagonism of Slow Reacting Substance in Anaphylaxis ( SRS-A) and Other Spasmogens on the Guinea Pig Tracheal
`Chain by Hydratropic Acids and Their Effects on Anaphylaxis . . . Margaret E. Greig* and Robert L. Griffin
`Antifertility Effects of Chlorine-Substituted Dioxolanes, Dithiolanes, and Dithianes in Male Rats
`. . . Allen F. Hirsch,* Kenneth C. Kolwyck, Larry A. Kraft, Roger E. Homm, and Do Won Hahn 116
`
`ll2
`
`Triphenylmethane Dyes as Inhibitors of Reverse Transcriptase, Ribonucleic Acid Polymerase, and Protein Synthesis.
`Structure-Activity Relationships . . . . . . . . . . . Lon-Lon Liao, Susan B. Horwitz, Mou-Tuan Huang,
`Arthur P. Grollman,* David Steward, and Jack Martin 117
`Catechol O-Methyltransferase. 5. Structure-Activity Relationships for Inhibition by Flavonoids
`. . . Ronald T. Borchardt* and Joan A. Huber 120
`
`BOOK REVIEWS
`The Hydrophobic Effect: Formation of Micelles and Biological Membranes. By Charles Tanford
`. . . Review by Irwin D. Kuntz, .Jr. 122
`
`Aleem, M.A., 108
`Aspinall, R. L., 50
`
`Borchardt, R. T., 120
`Bretting, C., 41
`Bruun, H., 41
`
`Cannon, ,J. G., 108, 110
`Carroll, F. I., 37
`Cashin, C. H., 53
`Cheng, C. C., 66
`Chheda, G. B., 79
`Clarke, R. L., 102
`Counsel!, R. E., 9, 16
`
`Danko, G., 1
`Daum, S. J ., 102
`Davis, A. L., 26
`del Prado, M ., 1
`Dunwell, D. W., 53
`
`El Masry, A.H., 16
`El Masry, S. E., 16
`Evans, D., 53
`
`Feit, P. W., 41
`Feller, D.R., 99
`
`Furukawa, H., 59
`
`Gage, L. D., 26
`Gilmore, W. F., 106
`Godfroid, .J .-,J ., 85
`Greig, M. E., 112
`Griffin, R. L:, 112
`Grimes, ,J. F., 26
`Grollman, A. P., 117
`Groves, W ., 90
`Gylys, ,J. A., 71
`-Hahn, D. W., 116
`Hannon, J., 1
`Hare, L. E., 16
`Hetnarski, B., 29
`Hicks, T. A., 53
`Hill, J.B., 50
`Hirsch, A. F., 116
`Homm, R. E., 116
`Hong, C. I., 79
`Horwitz, S. B., 117
`Hsu, S. Y., 20
`Huang, C. L., 20
`Huang, E.-S., 59
`Huang, M.-T., 117
`Huber, J. A., 120
`Huber, ,J. W ., III, 106
`
`AUTHOR INDEX
`Hulme, K. L., 26
`
`Jen,T., 90
`,Johnston, T. P., 104
`
`Kador, P. F., 99
`Kanno, M ., 34
`Khonje, P.R., 110
`Kim, S.-H., 59
`Kitchen, A., 53
`Kolwyck, K. C., 116
`Kraft, L.A., 116
`Kuntz, I. D., ,Jr., 122
`Kuriki, H., 34
`
`Labbe-Bois, R., 85
`Landon, M. J ., 1
`Larca, L. L., 1
`Laurelle, C., 85
`Lee, K.-H., 59
`Liao, L.-L., 117
`Loev, B., 90
`Long, .J.P., 108, 110
`Lutz, p,., 1
`
`Mann, T. A., 1
`Martin, J ., 117
`
`Matsumoto, J., 74
`McCaleb, G. S., 104
`McCord, T. J ., 26
`McLean, R. A., 90
`Merritt, W. V. P., 99
`Michel, I. M., 23
`Mikolajczak, K. L., 63
`Miller, D. D., 99
`Minami, S., 74
`Mitchell, G. N., 37
`Montgomery, .J. A., 104
`Mueller,N.J.,71
`Murata, T., 34
`
`Nielsen, 0. B. T., 41
`Nohara, A., 34
`
`O'Brien, R. D., 29
`
`Piantadosi, C., 59
`Powell, R. G ., 63
`
`Ray, R. E., 50
`Robertson, L. W ., 106
`Robinson, L. Q., 26
`
`Sanner, J. H., 9
`Sanno, Y ., 34
`Sierra, K., 9
`Smith, C.R., ,Jr., 63
`Smith, D. IL, 26
`Smith, R. V., 108
`Spivak, W ., 1
`Steward, D., 117
`
`Tozzi, S., 1
`
`Ukawa, K., 34
`
`Van Hoeven, H., 90
`Vashi, D., 1
`Venton, D. L., 9
`Villani, F. J., 1
`
`Wagner, H., 50
`Warawa, E. J., 71
`Waters, I. W., 20
`Wefer, E. A., 1
`Weinryb, I., 23
`Winters, D. W., 26
`
`Saijo, T., 34
`
`Zee-Cheng, R. K.-Y., 66
`
`There is no supplementary material for this issue.
`
`* In papers with more than one author, the asterisk indicates the name of the author to
`whom inquiries about the paper should be addressed.
`
`Thi:s material was Hipied
`
`Ex. 1091-0004
`
`

`

`104 Journal of Medicinal Chemistry, 1975, Vol. 18, No. l
`
`;]Pvv'l· 7S
`
`Notes
`
`Cocaine hydrochloride produced a marked numbness. ti-Cocaine
`hydrochloride produced essentially no effect.
`ti-Cocaine was evaluated in parallel with cocaine in the gt:inea
`pig intradermal wheal test as described by Bulbring and Waida.6
`The average threshold anesthetic concentration (TAC5) was ob(cid:173)
`tained from the dose-effect curve (semilogarithmic plot of dura(cid:173)
`tion in minutes vs. dosage) as described by Luduena and Hoppe.8
`Experimental values are furnished in Table I. ti-Cocaine was ap(cid:173)
`proximately one-third as active as cocaine.
`
`Ackno~ledgment. We thank Dr. Eugenio F. Bogado for
`the analgesic testing done on guinea pigs.
`
`References
`
`(1) R. Willstiitter, Chern. BeJ., 29, 2216 (1896).
`(2) R. Willstiitter, Chern. Ber., 31, 1534 (1898).
`(3) A. Heusner, Z. Naturforsch. l3, 12,602 (19,57).
`(4) R. Foster and H. R. Ing, J. Chern. Soc., 938 (1956).
`(5) R. Foster, H. R. Ing and V. Varagic, Brit. J. Pharmacol. Che(cid:173)
`mother., 10,436 (1955).
`(6) E. Bulbring and I. Wajda, J. Pharrnacol. Exp. Ther., 85, 78
`(1945).
`(7) C. A. Brown, J. Arner. Chem. Soc., 95,982 (1973).
`(8) F. P. Luduena and J. 0. Hoppe,.]. Pharmacol. Exp. Ther.,
`104, 40 (1952).
`
`Synthesis of Chlorozotocin, the 2-Chloroethyl Analog of the Anticancer
`Antibiotic Streptozotocin
`
`Thomas P. Johnston,* George S. McCaleb, and ,John A. Montgomery
`
`Kettering-Meyer Laboratory, Southern Research Institute, Birmingham, Alabama 35205. Received August 12, 1974
`
`Streptozotocin ( l) is a natural glucose-substituted N(cid:173)
`methyl-N-nitrosourea,1 a broad-spectrum antibiotic,2 and
`an experimental anticancer agent3 that has shown diabe(cid:173)
`togenic activity in animals4 and clinical activity in the
`treatment of malignant insulinomas in man. 5 Prior synthe(cid:173)
`sis of congeners of l has emphasized variation of the glucose
`moiety, 6-9 which resulted in enhancement of activity against
`leukemia L1210 in some cases judged on a comparable
`basis;9 limited testing of the ethyl and butyl analogs of 1
`indicated inactivity.3 A moderate increase in the antileu(cid:173)
`kemic activity of 1 was observed when the diabetogenic
`activity was suppressed with nicotinamide. 3 •10 The struc(cid:173)
`ture and properties of l suggested some time ago the re(cid:173)
`placement of the methyl group with a 2-chloroethyl group,
`a modification demonstrated11 to enhance markedly the
`effectiveness of a number of N-methyl-N-nitrosoureas
`against leukemia L1210.
`The principal previous attempt to synthesize 2-[3-(2-
`(:3,
`chloroethyl)-3-nitrosoureido ]-2-deoxy-D-glucopyranose
`"chlorozotocin") involved deacetylation of the tetraace(cid:173)
`tate ,1,12 a method modeled after the original synthesis of
`11 which has since been superseded by other methods.13 •14
`Interest in the synthesis of :3 was recently revived in view
`of the observation that, in mice, the tetraacetate 4, like
`J,15 showed reduced bone-marrow toxicity and was, unlike
`l, nondiabetogenic.16 Myelosuppression has been reported
`to be the limiting toxicity in the clinical use of the N-(2-
`chloroethyl)-N-nitrosoureas BCNU17
`[N,N' -bis(2-chlo(cid:173)
`roethyl)-N-nitrosourea] and CCNU18 [N-(2-chloroethyl)(cid:173)
`N'-cyclohexyl-N-nitrosourea, 5a].
`
`12~:~11
`HtL(·v
`I y
`
`NHCONR
`
`l,R=Me;Y= NO
`2,R = (CH2) 2CJ; Y = H
`3, R = (CH2) 2CJ; Y = NO
`
`CH20Ac
`
`OAc
`
`AcO
`
`NHCON(CH,).,CI
`I
`--
`NO
`
`4
`
`Chemistry. The (2-chloroethyl)urea 2 required for the
`synthesis of 3 was prepared by the addition of 2-chloroeth(cid:173)
`yl isocyanate to D-glucosamine in water and later in anhy(cid:173)
`drous N,N-dimethylformamide (DMF). The latter yield-
`
`improving variation was tried as a model for similar con(cid:173)
`versions in which hydrolysis of the isocyanate would be a
`significant competitive reaction in an aqueous system.
`The liberation of D-glucosamine from its hydrochloride
`with a basic ion-exchange resin was also a convenient in(cid:173)
`novation.
`The nitrosation of 2 with dinitrogen trioxide (chosen, as
`in the preparation of 1, 14 to simplify the separation of
`water-soluble :J) failed in a number of attempts with con(cid:173)
`ventional media: (1) in formic acid which would direct
`nitrosation to the desired position,1~ unwanted formyla(cid:173)
`t~on (of presumably the primary hydroxyl group and pos(cid:173)
`sibly others) occurred; (2) in water, the preferred medium
`for the preparation of 1,14 and in dilute formic acid, the
`reaction was exceedingly slow and incomplete after sever(cid:173)
`al hours; (3) in dilute hydrochloric acid, the reaction was
`somewhat faster but gave at least two products as indicat(cid:173)
`ed by tic. In concentrated hydrochloric acid, the reaction
`was essentially complete in less than 1 hr; recrystalliza(cid:173)
`tion of the resulting precipitate produced an analytically
`pure sample of 3. The pmr spectrum showed no evidence
`of random nitrosation and indicated a predominance of
`the a anomer. In aqueous solution, however, anomers of 3.
`would be expected to equilibrate as do various lots of
`crystalline 1.1 ,6 In a typical subsequent run, dinitrogen
`trioxide was introduced at a moderately fast rate until the
`reaction solution became red and intermittently thereafter
`until an appreciable amount of :i had precipitated; precip(cid:173)
`itation was completed by the addition of a proportionately
`large volume of ether. Unrecrystallized :1 was suitable
`(melting point, ir, and tic) for biological testing.
`The enhanced rate of nitrosation in concentrated hydro(cid:173)
`chloric acid was attributed to in situ generation of nitrosyl
`chloride in analogy to the standard preparation involving
`the addition of a concentrated aqueous solution of sodium
`nitrite to concentrated hydrochloric acid.in The observe~
`selective nitrosation was surpising, however, until the ni(cid:173)
`trosation of N-(2-chloroethyl)-N' -(trans-4-methylcyclohex(cid:173)
`yl)urea (Ii) under the same conditions (except that ethanol
`was added for solubilization) was demonstrated to give a
`nitrosourea that was identical with homogeneous samples
`of MeCCNU (7). Furthermore, a 1-hr treatment of a 2:1
`mixture of 5a and isomeric 5b with ethanolic concentrated
`hydrochloric acid resulted in a high-yield transformation
`to a nitrosourea that was very nearly identical with homo(cid:173)
`geneous samples of CCNU. These results parallel the se(cid:173)
`lective nitrosations and nitroso group migrations previous-
`
`Th is material was co,j:}ied
`at the NLM and may bE
`
`Ex. 1091-0005
`
`

`

`Journal of Medicinal Chemistry, 1975, Vol. 18, No. 1 105
`
`'l.'ahle I. Activity against Leukemia L121Q<l
`
`Diluent
`
`Dosage, b
`mg/kg/dose
`
`Schedule
`
`Median
`day of
`60-day
`survivors/total death
`
`% ILS
`
`Cell kill
`
`Citrate buffer
`Citrate buffer
`Citrate buffer
`
`CMCC
`Saline
`
`50
`7.5
`2.5
`
`20
`30
`
`qd 1-9
`qd 1-9
`qd 1-9
`
`Day 1 only
`Day 1 only
`
`0/10
`1/10
`0/10
`
`4/10
`9/10
`
`11. 0
`14. 5
`15.0
`18.0
`8.5
`17.0
`
`31
`36
`63
`
`100
`
`43%/dose
`47%/dose
`67%/dose
`
`cc:5 logs
`cc:6 logs
`
`Compd
`
`Control
`1
`4
`3
`Control
`4
`3
`
`nBDF 1 mice inoculated with 1Q5 Li210 cells intraperitoneally (ip) and treate_d i?. bM_aximum nontoxic dose as judged by concurrent
`toxicity controls in normal mice. cCarboxymethylcellulose sodium salt, 0.4% solution m saline.
`
`~y observed in essentially anhydrous formic acid11 •12 and
`1llustrate for the first time these effects in an aqueous sys(cid:173)
`tem. Thus, rearrangement may occur by abstraction of ni(cid:173)
`troso groups as nitrosyl chloride in one case and nitrosyl
`formate (formyl nitrite) in the other, both a source of ni(cid:173)
`trosonium ion for renitrosation in a more stable position.
`
`0
`
`NHCON(CH2).,Cl
`
`i -
`
`o-NCON(Cff,)2Cl
`
`-
`
`I
`I
`y z
`Sa, Y = H; Z = NO
`b,Y= NO;Z=H
`
`vision of Southern Research Institute undet the direction of Dr.
`W.R. Laster, Jr.
`2-[3-(2-Chloroethyl)ureido]-2-deoxy-o-glucopyranose (2). A.
`Preparation in Water. A cold (5°), stirred solution of {:l-D-glu(cid:173)
`cosamine22 [mp 110-111° (lit.23 mp 110-111°); 4.00 g, 22.3 mmol]
`in H2O (25 ml) was treated slowly with 2-chloroethyl isocyanate
`(1.97 ml, 22.3 mmol; Eastman Kodak Co.). After the addition,
`the reaction solution was stirred at 5° for 1 hr. The white precipi(cid:173)
`tate that formed was collected, washed with two 5-ml portions of
`EtOH, and dried overnight in vacuo over P2O5: yield of 2 4.50 g
`(71%); mp 160-161° dee; ir (KBr) 3600-3000 (OH, NH), 3000-2800
`(CH), 1620 (C=O), 1590 cm- 1 (CNH, amide II); homogeneous by
`tic (silica gel, 3:1 CHCla-MeOH, charring after NH4SO4-H2SO4
`spray).Anal. (C9H11CIN2O5) C, H, N.
`B. Preparation in DMF. A stirred suspension of {:l-o-glucosa(cid:173)
`mine (300 mg, 1.67 mmol) in DMF (4 ml) was treated with
`Cl(CH2hNCO (0.144 ml, 1.67 mmol), all the suspended solid
`having dissolved after -15 min. The solution was stirred for 1 hr,
`during which time a precipitate formed. The suspension was di(cid:173)
`luted with ether (40 ml), stirred for 30 min, and chilled. The
`product was collected, washed successively with ether (two 20-ml
`portions), EtOH (two 10-ml portions), and again ether (two 20-ml
`portions), and dried overnight in vacuo over P2O5: yield 450 mg
`(94%); mp 159-160° dee; ir identical with that of the analytically
`pure sample described above.
`o-Glueosamine (by Ion Exchange). A stirred solution of D-glu(cid:173)
`cosamine hydrochloride (1.00 g, 4.64 mmol) in H2O (25 ml) was
`treated at intervals with Dowex 1-X2 (OH-) ion-exchange resin
`until the pH was 9-10. The resin was removed by filtration and
`washed with two 10-ml portions of H2O; the filtrate and washings
`were combined and evaporated to dryness under reduced pressure
`with the aid of added EtOH (20 ml) and then ether (20 ml). The
`residual D-glucosamine was further dried in vacuo over P2O5
`[yield 650 mg (78%), mp -98° dee] and converted, in H2O, to 2,
`mp 159-160° dee, in 57% yield (ir identical with that of the ana(cid:173)
`lytical sample of 2).
`2- [ 3-( 2-C h I oroe thyl )-3-nitrosoureido ]-2-deoxy-o-glucopyra(cid:173)
`nose (3). N2O3 was bubbled into a cold (0-5°), stirred solution of
`2 (4.60 g, 16.2 mmol) in concentrated HCl (46 ml) at a moderate(cid:173)
`ly fast rate until the solution became red and then intermittently
`until an appreciable amount of solid had formed ( - 1.25 hr).
`Ether (400 ml) was added and the mixture was stirred at 0° for 20
`min. The precipitate was collected on a filter, washed with two
`20-ml portions of ether, and dried overnight in vacuo over P2O5:
`yield 2.67 g (53%); mp 140-141° dee. This sample was identical
`(melting point, ir, and tlc) with the analytical sample obtained
`from a previous, 45-min nitrosation and recrystallized from
`EtOH: ir (KBr) 3600-3100 (OH, NH), 3000-2800 (CH), 1695
`(C=O), 1540 (CNH, amide II), 1490 cm- 1 (N=O); pmr (DMSO(cid:173)
`d6-TMS, chemical shifts quoted as either ranges or approximate
`centers) o 3.0-3.9 [m, CH2OH, H-5, H-4, H-3, H-2, overlapping
`upfield half of A2B 2 system due to N(CH2hCl], 4.12 [pseudo t
`from downfield half of A2B2 system due to N(CH2hCl], 3.9-5.2 (3
`OH's, apparently exchanging with small amount of H2O in sol(cid:173)
`vent), 4.67 (d, J 1 ,2 "" 9 Hz, H-1 of f:l anomer;2·1 obscured by OH's
`until after addition of D2O), 5.14 (d, J1,2 "" 2 Hz, H-1 of a anom-
`er;U coupling measured after addition of D2O, which eliminated
`coupling with OH), 6.6 (br m, OH probably at C-1), 7.8 (br d, NH
`tThe log P value of 3, as determined by Dr. W. ,J. Haggerty of Midwest
`of a anomer), 8_5 (br d, NH of f:l anomer). [Pmr indicated an a:{:l
`Research Institute, Kansas City, from the octanol-water partition coeffi-
`cient and transmitted by Dr. R. R. Engle of Drug Research and Develop-
`anomeric ratio of -10: 1, but 3 appeared homogeneous by tic on
`ment, National Cancer Institute, is - 1.02. Reported" values for other
`silica gel with 5: 1 CHC'3-MeOH and detection by uv and char-
`streptozotocin analogs range from -0.82 to -1.57 (streptozotocin -1.-15).
`Th is mate ri a I was co,pied
`atth,a NLM and may be
`~ubjed US Copyright Laws
`
`CHa
`
`6,Y=H
`7,Y= NO
`
`Biologic Evaluation. The prediction of superior activi(cid:173)
`ty for chlorozotocin (3) against leukemia L1210 has been
`corroborated by experiment (see Table I). Groups of leu(cid:173)
`kemic mice· were treated daily for 9 days with 3, its te(cid:173)
`traacetate 4, and streptozotocin ( 1), respectively, in a sin(cid:173)
`?ie experiment. On this schedule, the maximum increase
`in life span; based on the median day of death, produced
`by 3 was more than twice that produced by either of the
`other two nitrosoureas, proving the clear superiority of 3.
`The schedule used in this experiment (qd 1-9) was chosen
`because it is the optimal schedule for 1,20 but it is not op(cid:173)
`timal for 3 or 4. In another single experiment on the opti(cid:173)
`mal schedule (single dose) for 3 and 4, 3 killed 6 logs or
`greater of L1210 cells, curing nine of ten animals; 11 killed
`about 5 logs and cured four animals, increasing the life
`span of the dying animals 100%. Given in this way 1 is
`said to be inactive. 20 In addition to its high-level L1210
`activity, 3 showed reduced bone-marrow toxicity21 and
`has the advantage of water solubility over 4.t On the basis
`of activity and low bone-marrow toxicity, 3 is now under-.
`going pharmacologic evaluation preliminary to clinical
`trials.
`
`Experimental Section
`
`Melting points with a range were determined with a Mel-Temp
`apparatus and are uncorrected; those without range, with a Ko(cid:173)
`fler Heizbank. Ir spectra were determined with Perkin-Elmer 521
`and 621 spectrophotometers and pmr spectra with a Varian XL-
`100-15 spectrometer. Analytical results indicated by element
`symbols were within ±0.4% of the theoretical values. Elemental
`analysis of 3 was performed by Galbraith Laboratories, Knoxville,
`Tenn.; spectral determinations and elemental analysis of 2 were
`performed in the Molecular Spectroscopy Section of Southern Re(cid:173)
`search Institute under the direction of Dr. W. C. Coburn, Jr.; and
`biological evaluation was performed in the Cancer Screening Di-
`
`Ex. 1091-0006
`
`

`

`10(; Journal of Medicinal Chemistry, 1975, Vol. 18, No. 1
`
`Notes
`
`ring after (NH.1hSOrH2SO4 spray.] Anal. (C9H16CJN3O1) C, H,
`N.
`Nitrosation of N-(2-Chloroethyl)-N' -(trans-4-methylcyclo(cid:173)
`hexyl)urea ((;) in Ethanolic Hydrochloric Acid. N2O3 was bub(cid:173)
`bled into a cold (0-5°), stirred solution of (;12 (125 mg, 0.570
`mmol) in concentrated HCI (2 ml) and EtOH (1 ml) at a moder(cid:173)
`ate rate for 15 min. Kept at 0-5°, the reaction mixture was stirred
`for 5 min before and after dilution with cold H2O (20 ml). The
`light-yellow MeCCNU (7) that separated was collected and dried
`in vacuo over !'20 5 : yield 105 mg (74%); mp 70° (lit. 12 mp 70°);
`identity with an authentic sample established by ir (KBr) and
`pmr (CDCl3-TMS).
`Nitroso Group Migration in Ethanolic Hydrochloric Acid:
`CCNU (5a). An isomeric mixture 11 (200 mg), mp 69° dee, of 5a
`(-65%) and 5b (-35%) was stirred in suspension in a cold (0-5°)
`solution of EtOH (2 ml) in concentrated HCl (3 ml) for 1 hr. The
`mixture was diluted with cold H2O (20 ml) and stirred at 0° for
`20 min more. The light-yellow solid was collected, washed with
`cold H 2O (5 ml), and dried in vacuo over P2(h: yield 190 mg
`(95%); mp 89° (lit_ll mp of CCNU 90°); pmr (CDCla-TMS)
`showed possibly a trace of 5b.
`
`Acknowledgment. This investigation was supported by
`the Division of Cancer Treatment, National Cancer Insti(cid:173)
`tute, National Institutes of Health through Contract No.
`NOl-CM-33712. The authors are indebted to Dr. Philip S.
`Schein for encouraging discussions and to Mrs. Martha C.
`Thorpe for interpretation of pmr spectra.
`
`References
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`
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`
`Synthesis and Antimicrobial Evaluation of N-n-Alanyl-1-aminoethylphosphonic Acid
`
`,Joseph W. Huber, III, W. Franklin Gilmore,*
`
`Department of Medicinal Chemistry
`
`and Larry W. Robertson
`Department of Pharmacognosy, School of Pharmacy, University of Mississippi, University, Mississippi 38677. Received June 13, 1974
`
`0
`II
`CH,CHCNHCHCO.,H
`, I
`I
`-
`NH2
`CH3
`
`2
`
`0
`II
`CHiCH CNHCHP03H2
`I
`I
`CHi
`1
`
`NH2
`
`A number of antibiotic substances are known to exert
`antimicrobial activity by virtue of their ability to inhibit
`the synthesis of bacterial cell wall material.1 Some of the
`compounds incl