CA2248598C - Hydroxyamidino derivatives useful as nitric oxide synthase inhibitors - Google Patents

Abstract

The current invention discloses hydroxyamidino derivatives useful as nitric oxide synthase inhibitors as defined by Formula I:
(see formula I) Substituents A, X and R1 to R8 are as defined in the application.

Description

gYD$OYYAMIDI1Q0 ~IVA'1'IVE6 11R8F~LlL 11R
~1ITR~I~ ~ IS~1IHITORB
$ack~ncl ~~ the Invention yield of the zRventian The present iav$atian relates to hydxoxyamidino derivatives and their use in therapy, 3n particular t3leir 1p use as nitric oasde synthaae inhibitors and is a continuation.-in-part of n.5. patent No. 5,945,408 issued August 31.. 1999.

Related 1A~
=t has been kncnan since the early 1980~s that the yascnlar re~.aration bxouqht about by acetylarioliae a.s 20 dependent on the preseaas of the endotheliam sad this activity wt~ss ascribed to a labile humoral factor termed e~,dat~lium-derived relaxing factor (EDRF). The activity er nitric oxide (po) as a vasodilator has bass known far we~.i aver 100 y8ars and No i~a the active component of 25 amyl nitrite, glyaeryltarinitrate and other nitravagadilataX;a. The recent ident9.ficataon of EDIiF as g0 has ec~a.ncaded With the discovery of a biaehemical pathwe~y Dy h No ~.s synthesized Pram the amino acrd 1,...aacgini.ne by the enzyme NO syuthase.

~O is the endageaous stimulator of the soluble guanxlate cyclase and is involved in a number of bj.,ological actions 3n addition to endatheliuu~dependent relaxation including cytotoxicsty of phagocytic cells and 35 cell-to-cell communisation in. the centxal nervous system (see l~oncada et a1. siochemxaal pharnvacolosrv~ 38, 1749-115 ( 1989 ) dvr~d Mo coda et a1. Pharmacolo ical Reviews .
113, 1b9-142 (1991). It is now thought that excess Po production may be involved in a number of conditions, particularly conditions which involve systemic hypotension such as toxic shock and therapy with certain cytokines.
The synthesis of NO from L-arginine can be inhibited by the L-arginine analog, L-N-monomethyl-arginine (L-N1~IA) and the therapeutic use of L-NMMA for the treatment of toxic shock and other types of systemic hypotension has been proposed (WO 91/04024 and GB-A-2240041). The therapeutic use of certain other NO synthase inhibitors apart from L-NMMA for the same purpose has also been proposed in WO 91/04024 and in EP-A-0446699.
It has recently become apparent ~Ehat there are at least three types of NO synthase as follows:
(i) a constitutive, Ca++/calmodulin dependent enzyme, located in the endothelium, that releases NO in response to receptor or physical stimulation.
(ii) a constitutive, Ca++/calmodulin dependent enzyme, located in the brain, that releases NO in response to receptor or physical stimulation.
(iii) a Ca++ independent enzyme which is induced after activation of vascular smooth muscle, macrophages, endothelial cells, and a number of other cells by endotoxin and cytokines. Once expressed this inducible NO synthase synthesizes NO for long periods.
The NO released by the constitutive enzymes acts as a transduction mechanism underlying several physiological responses. The NO produced by the inducible enzyme is a cytotoxic molecule for tumor cells and invading microorganisms. It also appears that the adverse effects of excess NO production, in particular pathological vasodilation and tissue damage, may result largely from the effects of NO synthesized by the inducible NO
synthase.
.. SUBSTITUTE SHEET (RULE 26) There is also a growing body of evidence that NO may be involved in the degeneration of cartilage which takes place in certain conditions such as arthritis and it is also known that NO synthesis is increased in rheumatoid arthritis. Accordingly, further conditions in which there is an advantage in inhibiting NO production from L-arginine include auto immune and/or inflammatory conditions affecting the joints, for example arthritis, inflammatory bowel disease, cardiovascular ischemia, diabetes, congestive heart failure, myocarditis, atherosclerosis, migraine, reflux esophagitis, diarrhea, irritable bowel syndrome, cystic fibrosis, emphysema, hyperalgesia (allodynia), cerebral ischemia (both focal ischemia, thrombotic stroke and global-ischemia, secondary to cardiac arrest), other central nervous system disorders mediated by NO and other disorders mediated by NO including opiate tolerance in patients needing protracted opiate analgesics, and benzodiazepine tolerance in patients taking benzodiazepines, and other addictive behavior, for example, nicotine and eating disorders.
Further conditions in which there is an advantage in inhibiting NO production from L-arginine include systemic hypotension associated with septic and/or toxic shock induced by a wide variety of agents; therapy with cytokines such as TNF, IL-1 and IL-2; and as an adjuvant to short term immunosuppression in transplant therapy.
Some of the NO synthase inhibitors proposed for therapeutic use so far, and in particular L-NMMA, are non-selective in that they inhibit both the constitutive and the inducible NO synthase. Use of such a non-selective NO synthase inhibitor requires that great care be taken in order to avoid the potentially serious consequences of over-inhibition of the constitutive NO-SUBSTITUTE SHEET (RULE 26) synthase ~.ncludang hyperr~nsion and possib7.e thro~mboais and tissue damage. In particular, a.n the case of the therapeutic nee of L-Nt~lA fox the treatment of toxic shocx it has been reco~dended that the patient raust be subject to continuous bland pressure monitoring throughout the treatment. Thus, while nvn-selective NO
syathaae inhibitors Have therapQUtic utility provided that appropriate precautions are taken, Na aynthase inhibitors whj.eh are selsct~.ve in the sense that they inhabit thg inducible ND.synthase to a oonaiderably greater extent,than the constitutive isotorms of No synthaae would be of even greater therapeutic benefit and easier to use.
DPb34/12155, W094114780, W093/1305b, ~P04~6G99A1 and U.B. Patent No. 5,19Z,~453 disclose compounds that inhibit nitric aside synthesis and preferentially ~.nhibit the induaible isofora~ of nitric oxide spnthase.
Summary of the Inyention In a broad asgeet, the present imrentian is directed to inhybiting or modulating nitric oxide synthesis in a subject in need of euah inhibition or ~m~odulation by administering a compound which preferentially :i.nhibits or modulates the inducible isoform of nitric oxide synthase over the constitutive isofaxms of n~.tric oxide spnthase.
It is also another object of th~ present imreation tn lower nitric ox.~i.de levels in a sub jest in need of such laweri.ng .
Compounds of the present invention are represented by the following chemical fcFmula:

~N O
X
R2 ~ N~ ~A-R3 Rs Rl \NR~Rg (I) and pharmaceutically acceptable salts, wherein:
A is selected from 0 (oxygen) or S and may be taken together with R4 to form a heterocyclic ring; or i5 A is N when R3 and R~ are taken together to form a heterocyclic ring; or R5 and A-R3 are taken together to form a covalent bond; or A is N-R3 provided R3 is not a heterocyclic radical;
R1 is not present or is selected from the group consisting of hydrogen, hydroxyalkyls, alkoxyalkyls, lower alkyls and haloalkyls;

R2 is selected from the group consisting of straight and branched lower alkyls, lower alkenyls, and lower alkynyls, cycloalkyls, cycloalkenyls, haloalkyls;
R3 is selected from the group consisting of aryl, heteroaryl, alkylaryl, alkylheteroaryl, all optionally substituted by one or more of halogen, nitrile, carboxy, carboxyalkyl, carboxyalkylaryl; or R3 is selected from the group consisting of H, alkyl, alkenyl, CH20C(=O)YR6, alkylhydroxy, alkylpolyhydroxy, alkyl(poly)oxyacyl, alkylcarboxy, optionally substituted .. SUBSTITUTE SHEET (RULE 26) by one or more of alkyl, hydroxy, amino, carboxy, carboxyalkyl, alkylcarbonyl;
R4 is selected from H, OH, SH, OR6, SR6, OC(=O)R6, SC(=O)R6, CH20C(=O)YR6, OC(=O)YR6, SC(=0)YR6;
Y is independently selected from O, S, CH2, CHR6, C(R6)2, NH, NR6;
R5 is selected from H, OH, SH, OR6, SR6, OC(=O)R6, SC(=O)R6, CH20C(=O)YR6, OC(=O)YR6, SC(=0)YR6;
provided that R4 and R5 are not simultaneously hydrogen or CH20C(=O)YR6.
R6 is selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, heteroaryl all optionally substituted by one or more alkyl, alkoxy, hydroxy, halogen, trifluoromethyl, nitro, cyano, or amino groups;
R~ is selected from H, S(O)R9, S02R9, CH20C(O)-R9~ C(0)-R9 where C(O)-R9 can represent natural and synthetic amino acids or R9 can be defined as below, or R~ and R3 taken together comprise a 5- or 6- membered heterocyclic ring containing two or more heteroatoms, optionally substituted with alkyl and/or oxygen functions including carbonyl, or taken together comprise a metal complex containing a divalent cation, or a boron complex;
R8 is selected from H, acyl;
R9 is selected from substituted dihydropyridyl, alkyl, thioalkoxy, alkoxy, amino, cycloalkoxy, optionally substituted with one or more of amino, alkyl, alkylaryl, heteroaryl, alkylheteroaryl, alkylmercaptoalkyl, which may optionally be substituted with one or more of hydroxy, amino, guanidino, iminoalkyl;
SUBSTITUTE SHEET (RULE 26) X is selected from the group consisting of lower alkylenes, lower alkenylenes and lower alkynylenes and which may optionally be substituted by one or more alkyl, alkoxy, hydroxy, halogen, trifluoromethyl, nitro, cyano, or amino groups; or X is selected from the group consisting of the formula -(CH2)kQ(CH2)t- where k is 1, 2 or 3, t is 1, 2 or 3 and Q is O (oxygen), Se, SiE2 where E is lower alkyl, aryl, S(O)g where g is 0, 1 or 2, or NR where R is H or lower alkyl which may be optionally substituted with lower alkyl, lower alkoxy, hydroxy, halogen, trifluoromethyl, nitro, cyano, amino; or X is selected from the group consisting of the formula -(CH2)mT(CH2)n- where m is 0, 1 or 2, n is 0, 1 or 2, T
is a 3 to 6 membered carbocyclic or heterocyclic ring, aromatic ring or heteroaromatic ring which may optionally be substituted by one or more substituents selected from the group consisting of lower alkyl, lower alkoxy, hydroxy, halogen, nitro, cyano, trifluoroalkyl and amino.
It is an object of the present invention to provide compounds that have usefulness as inhibitors of nitric oxide synthase. These compounds also preferentially inhibit the inducible form over the constitutive forms.
DETAILED DESCRIPTION OF THE INVENTION

~N O
X
R2 ~ N~ ~A-R3 3o R5 Rl NR~Rg (I) SUBSTITUTE SHEET (RULE 26) and pharmaceutically acceptable salts, wherein:
A is selected from 0 (oxygen) or S and may be taken together with R4 to form a heterocyclic ring; or A is N when R3 and R~ are taken together to form a heterocyclic ring; or R5 and A-R3 are taken together to form a covalent bond; or A is N-R3 provided R3 is not a heterocyclic radical;
R1 is not present or is selected from the group consisting of hydrogen, hydroxyalkyls, alkoxyalkyls, lower alkyls and haloalkyls;
R2 is selected from the group consisting of straight and branched lower alkyls, lower alkenyls, and lower alkynyls, cycloalkyls, cycloalkenyls, haloalkyls;
R3 is selected from the group consisting of aryl, heteroaryl, alkylaryl, alkylheteroaryl, all optionally substituted by one or more of halogen, nitrite, carboxy, carboxyalkyl, carboxyalkylaryl; or R3 is selected from the group consisting of H, alkyl, alkenyl, CH20C(=0)YR6, alkylhydroxy, alkylpolyhydroxy, alkyl(poly)oxyacyl, alkylcarboxy, optionally substituted by one or more of alkyl, hydroxy, amino, carboxy, carboxyalkyl, alkylcarbonyl;
R4 is selected from H, OH, SH, OR6, SR6, OC(=0)R6, SC(=O)R6, CH20C(=O)YR6, OC(=0)YR6, SC(=O)YR6;
Y is independently selected from O, S, CH2, CHR6, C(R6)2.
NH, NR6;
SUBSTITUTE SHEET (RULE 26) R5 is selected from H, OH, SH, OR6, SR6, OC(=0)R6, SC(=O)R6, CH20C(=O)YR6, OC(=O)YR6, SC(=O)YR6;
provided that R4 and R5 are not simultaneously hydrogen or CH20C(=O)YR6.
R6 is selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, heteroaryl all optionally substituted by one or more alkyl, alkoxy, hydroxy, halogen, trifluoromethyl, nitro, cyano, or amino groups;
R~ is selected from H, S(0)R9, S02R9, CH20C(O)-R9~ C(O)-R9 where C(O)-R9 can represent natural and synthetic amino acids or R9 can be defined as below, or R~ and R3 taken together comprise a 5- or 6- membered heterocyclic ring containing two or more heteroatoms, optionally substituted with alkyl and/or oxygen functions including carbonyl, or taken together comprise a metal complex containing a divalent cation, or a boron complex;
R8 is selected from'H, acyl;
R9 is selected from substituted dihydropyridyl, alkyl, thioalkoxy, alkoxy, amino, cycloalkoxy, optionally substituted with one or more of amino, alkyl, alkylaryl, heteroaryl, alkylheteroaryl, alkylmercaptoalkyl, which may optionally be substituted with one or more of hydroxy, amino, guanidino, iminoalkyl;
X is selected from the group consisting of lower alkylenes, lower alkenylenes and lower alkynylenes and which may optionally be substituted by one or more alkyl, alkoxy, hydroxy, halogen, trifluoromethyl, vitro, cyano, or amino groups; or X is selected from the group consisting of the formula -(CH2)kQ(CH2)t- where k is 1, 2 or 3, t is 1, 2 or 3 and SUBSTITUTE SHEET (RULE 26) Q is O (oxygen), Se, SiE2 where E is lower alkyl, aryl, S(O)g where g is 0, 1 or 2, or NR where R is H or lower alkyl which may be optionally substituted with lower alkyl, lower alkoxy, hydroxy, halogen, trifluoromethyl, 5 nitro, cyano, amino; or X is selected from the group consisting of the formula -(CH2)mT(CH2)n- where m is 0, 1 or 2, n is 0, 1 or 2, T
is a 3 to 6 membered carbocyclic or heterocyclic ring, 10 aromatic ring or heteroaromatic ring which may optionally be substituted by one or more substituents selected from the group consisting of lower alkyl, lower alkoxy, hydroxy, halogen, nitro, cyano, trifluoroalkyl and amino.
A preferred embodiment of the present invention is a compound of the formula (I):

~N O
X
R2 ~ N~ ~A-R3 Rs Rl \NR~Rg (I) and pharmaceutically acceptable salts, wherein:
A is selected from O (oxygen) or S and may be taken together with R4 to form a heterocyclic ring; or A is N when R3 and R~ are taken together to form a heterocyclic ring; or R5 and A-R3 are taken together to form a covalent bond; or A is N-R3 provided R3 is not a heterocyclic radical;
SUBSTITUTE SHEET (RULE 26) WO 97/32844 ~ PCT/US97/03282 R1 is selected from the group consisting of hydrogen, hydroxyalkyls of from 1 to 4 carbon atoms, alkoxyalkyls of from 1 to 4 carbon atoms in each position, lower alkyls of from 1 to 8 carbon atoms and haloalkyls of from 1 to 4 carbon atoms;
RZ is selected from the group consisting of straight and branched lower alkyls of from 1 to 4 carbon atoms, lower alkenyls and lower alkynyls of from 2 to 4 carbon atoms, cycloalkyls of from 1 to 4 carbon atoms, cycloalkenyls of from 3 to 8 carbon atoms, and haloalkyls of from 1 to 4 carbon atoms;
R3 is selected from the group consisting of aryl, heteroaryl, alkylaryl, alkylheteroaryl, all optionally substituted by one or more of halogen, nitrile, carboxy, carboxyalkyl, carboxyalkylaryl; or R3 is also selected from the group consisting of H, alkyl of from 1 to 4 carbon atoms, alkenyl of from 2 to 4 carbon atoms, CH20C(=O)YR6, alkylhydroxy, alkylpolyhydroxy, alkyl(poly)oxyacyl, alkylcarboxy, optionally substituted by one or more of alkyl of from 1 to 4 carbon atoms, hydroxy, amino, carboxy, carboxyalkyl, alkylcarbonyl;
R4 is selected from H, OH, SH, OR6, SR6, OC(=0)R6, SC(=O)R6, CH20C(=O)YR6, OC{=O)YR6, SC(=O)YR6;
Y is independently selected from O, S, CH2, CHR6, C(R6)2.
NH, NR6;
R5 is selected from H, OH, SH, OR6, SR6, OC(=O)R6, SC(=O)R6, CH20C(=0)YR6, OC(=O)YR6, SC(=O)YR6;
SUBSTITUTE SHEET (RULE 26) provided that R4 and R5 are not simultaneously hydrogen or CH20C(=0)YR6.
R6 is selected from hydrogen, alkyl of from 1 to 4 carbon atoms, alkenyl and alkynyl of from 2 to 4 carbon atoms, cycloalkyl of from 3 to 8 carbon atoms, heterocyclic of from 5 to 8 members, aryl, heteroaryl all optionally substituted by one or more alkyl of from 1 to 4 carbon atoms, alkoxy, hydroxy, halogen, trifluoromethyl, nitro, cyano, or amino groups;
R~ is selected from H, S(O)R9, S02R9, CH20C(O)-R9~ C(O)-R9 where C(O)-R9 can represent natural and synthetic amino acids or R9 can be defined as below, or R~ and R3 taken together comprise a 5- or 6- mem~ered heterocyclic ring containing two or more heteroatoms, optionally substituted with alkyl of from 1 to 4 carbon atoms and/or oxygen functions including carbonyl, or taken together comprise a metal complex containing a divalent cation, or a boron complex;
R8 is selected from H, acyl;
R9 is selected from substituted dihydropyridyl, alkyl of from 1 to 4 carbon atoms, thioalkoxy, alkoxy, amino, cycloalkoxy, optionally substituted with one or more of amino, alkyl of from 2 to 4 carbon atoms, alkylaryl, heteroaryl, alkylheteroaryl, alkylmercaptoalkyl, which may optionally be substituted with one or more of hydroxy, amino, guanidino, iminoalkyl;
X is selected from the group consisting of alkylenes, alkenylenes and alkynylenes having 2 to 6 carbon atoms and which may optionally be substituted by one or more alkyl groups; or SUBSTITUTE SHEET (RULE 26) WO 97!32844 PCTIITS97I03282 X is selected from the group consisting of the formula -(CH2)kQ(CH2)t- where k is 1, 2 or 3, t is 1, 2 or 3 and Q is 0 (oxygen), Se, SiE2 where E is lower alkyl, aryl, S(0)g where g is 0, 1 or 2, or NR where R is H or lower alkyl which may be optionally substituted with lower alkyl, lower alkoxy, hydroxy, halogen, trifluoromethyl, nitro, cyano, amino; or X is selected from the group consisting of the formula -(CH2)mT(CH2)n- where m is 0, 1 or 2, n is 0, 1 or 2, T
is a 3 to 6 membered carbocyclic or heterocyclic ring, aromatic ring or heteroaromatic ring which may optionally be substituted by one or more substituents selected from the group consisting of lower alkyl, lower alkoxy, hydroxy, halogen, nitro, cyano, triflfzoroalkyl and amino.
A preferred embodiment of the present invention is a compound of the formula (I):

~N O
X
R2 ~ N~ ~A-R3 2o Rs Rl \NR7Rg (I) and pharmaceutically acceptable salts, wherein:
A is selected from O (oxygen) or S and may be taken together with R4 to form a heterocyclic ring; or A is N when R3 and R~ are taken together to form a heterocyclic ring; or R5 and A-R3 are taken together to form a covalent bond; or A is N-R3 provided R3 is not a heterocyclic radical;
SUBSTITUTE SHEET (RULE 26) R1 is selected from the group consisting of hydrogen, hydroxyalkyls of from 1 to 4 carbon atoms, alkoxyalkyls of from 1 to 4 carbon atoms in each position, lower alkyls of from 1 to 8 carbon atoms and haioalkyls of from 1 to 4 carbon atoms;
R2 is selected from the group consisting of straight and branched lower alkyls of from 1 to 4 carbon atoms, lower alkenyls and lower alkynyls of from 2 to 4 carbon atoms, cycloalkyls of from 1 to 4 carbon atoms, cycloalkenyls of from 3 to 8 carbon atoms, and haloalkyls of from 1 to 4 carbon atoms;
R3 is selected from the group consisting of aryl, heteroaryl, alkylaryl, alkylheteroaryl, all optionally substituted by one or more of halogen, nitrile, carboxy, carboxyalkyl, carboxyalkylaryl; or R3 is also selected from the group consisting of H, alkyl of from 1 to 4 carbon atoms, alkenyl of from 2 to 4 carbon atoms, CH20C(=0)YR6, alkylhydroxy, alkylpolyhydroxy, alkyl(poly)oxyacyl, alkylcarboxy;
R4 is selected from H, OH, SH, OR6, SR6, OC(=O)R6, SC(=0)R6, CH20C(=O}YR6;
Y is independently selected from O, S, CH2, CHR6, C(R6)2.
NH, NR6;
R5 is selected from H, OH, SH, OR6, SR6, OC(=0)R6, SC(=O)R6, CH20C(=O)YR6;
provided that R4 and R5 are not simultaneously hydrogen.
R6 is selected from hydrogen, alkyl of from 1 to 4 carbon atoms, alkenyl and alkynyl of from 2 to 4 carbon atoms, cycloalkyl of from 3 to 8 carbon atoms, heterocyclic of SU9STITUTE SHEET (RULE 26) from 5 to 8 members, aryl, heteroaryl all optionally substituted by one or more alkyl of from 1 to 4 carbon atoms or hydroxy groups;
5 R~ is selected from H, S(0)R9, S02R9, CH20C(O)-R9~ C(O)-R9 where C(O)-R9 can represent natural and synthetic amino acids or R9 can be defined as below, or R~ and R3 taken together comprise a 5- or 6- membered heterocyclic ring containing two or more heteroatoms;
R$ is selected from H, acyl;
R9 is selected from substituted dihydropyridyl, alkyl of from 1 to 4 carbon atoms, thioalkoxy, alkoxy, amino, cycloalkoxy;
X is selected from the group consisting of alkylenes, alkenylenes and alkynylenes having 2 to 6 carbon atoms and which may optionally be substituted by one or more alkyl groups; or X is selected from the group consisting of the formula -(CH2)kQ(CH2)t- where k is 2 or 3, t is 1 or 2 and Q is O
(oxygen), Se, SiE2 where E is lower alkyl, aryl, S(O)g where g is 0, 1 or 2, or NR where R is H or lower alkyl which may be optionally substituted with lower alkyl; or X is selected from the group consisting of the formula -(CH2)mT(CH2)n- where m is 0, 1 or 2, n is 0, 1 or 2, T
is a 3 to 6 membered carbocyclic or heterocyclic ring, aromatic ring or heteroaromatic ring which may optionally be substituted by one or more substituents selected from the group consisting of lower alkyl, lower alkoxy, hydroxy, halogen, vitro, cyano, trifluoroalkyl and amino.
SUBSTITUTE SHEET (RULE 26) Another preferred embodiment of the present invention is a compound of the formula (I) and pharmaceutically acceptable salts; wherein:
A is selected from O (oxygen) and may be taken together with R4 to form a heterocyclic ring; or A when R5 and A-R3 are taken together forms a covalent bond;
A is N-R3 where the R3 radicals are selected from hydrogen, alkyl of from 1 to 4 carbon atoms or aryl;
R1 is selected from the group consisting of hydrogen, hydroxyalkyls of from 1 to 4 carbon atoms and lower alkyls of from 1 to 8 carbon atoms;
R2 is selected from the group consisting of straight and branched lower alkyls of from 1 to 4 carbon atoms and haloalkyls of from 1 to 4 carbon atoms;
R3 is selected from the group consisting of aryl, heteroaryl, alkylaryl, alkylheteroaryl, all optionally substituted by one or more of halogen, nitrile, carboxy, carboxyalkyl, carboxyalkylaryl; or R3 is also selected from the group consisting of H, alkyl of from 1 to 4 carbon atoms and alkenyl of from 2 to 4 carbon atoms;
R4 is selected from H, OH, SH, OR6, SR6;
R5 is selected from H, OH, SH, OR6, SR6;
provided that R4 and R5 are not simultaneously hydrogen.
SUBSTITUTE SHEET (RULE 26) R6 is selected from hydrogen, alkyl of from 1 to 4 carbon atoms, alkenyl and alkynyl of from 2 to 4 carbon atoms and cycloalkyl of from 3 to 8 carbon atoms;
R~ is selected from H and where C(O)-R9 can represent natural and synthetic amino acids;
R$ is selected from H and acyl;
X is selected from the group consisting of alkylenes, alkenylenes and alkynylenes having 2 to 6 carbon atoms and which may optionally be substituted by one or more alkyl groups; or X is selected from the group consisting of the formula -(CH2)kQ(CH2)t- where k is 2 or 3, t is 1 or 2 and Q is 0 (oxygen), S(O)g where g is 0, 1 or 2, or NR where R is H
or lower alkyl or X is selected from the group consisting of the formula -(CH2)mT(CH2)n- where m is 0, 1 or 2, n is 0, 1 or 2, T
is a 3 to 6 membered carbocyclic or heterocyclic ring, aromatic ring or heteroaromatic ring.
Another preferred embodiment of the present invention is a compound of the formula (I) and pharmaceutically acceptable salts; wherein:
A is 0 (oxygen);
Rl is hydrogen;
RZ is methyl;
R3 is selected from the group consisting of hydrogen, and lower alkyls of 1 to about 4 carbon atoms;
R4 is hydroxy;
SUBSTITUTE SHEET (RULE 26) R5 is hydrogen or hydroxy;
R~ is hydrogen;
R8 is hydrogen;
X is an alkylene having 3 to 5 carbon atoms.
The present invention includes compounds of formula (I} in the form of salts, in particular acid addition salts. Suitable salts include those formed with both organic and inorganic acids. Such acid addition salts will normally be pharmaceutically acceptable although salts of non-pharmaceutically acceptable salts may be of utility in the preparation and purification of the compound in question. Thus, preferred salts include those formed from hydrochloric, hydrobromic, sulphuric, citric, tartaric, phosphoric, lactic, pyruvic, acetic, succinic, fumaric, malefic, oxaloacetic, methanesulphonic, ethanesulphonic, p-toluenesulphonic, benzenesulphonic and isethionic acids. Salts of the compounds of formula (I) can be made by reacting the appropriate compound in the form of the free base with the appropriate acid.
While it may be possible for the compounds of formula (I) to be administered as the raw chemical, it is preferable to present them as a pharmaceutical composition. According to a further aspect, the present invention provides a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof, together with one or more pharmaceutically acceptable carriers thereof and optionally one or more other therapeutic ingredients. The carriers) must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.
SUBSTITUTE SHEET (RULE 26) WO 97/32844 ~ PCT/US97/03282 The formulations include those suitable for oral, parenteral (including subcutaneous, intradermal, intramuscular, intravenous and intraarticular), rectal and topical (including dermal, buccal, sublingual and intraocular) administration although the most suitable route may depend upon for example the condition and disorder of the recipient. The formulations may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy. All methods include the step of bringing into association a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof ("active ingredient") with the carrier which constitutes one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association the active ingredient with liquid carriers or ffinely divided solid carriers or both and then, if necessary, shaping the product into the desired formulation.
Formulations of the present invention suitable for oral administration may be presented as discrete units such as capsules, cachets or tablets each containing a predetermined amount of the active ingredient; as a powder or granules; as a solution or a suspension in an aqueous liquid or a non-aqueous liquid; or as an oil-in-water liquid emulsion or a water-in-oil liquid emulsion.
The active ingredient may also be presented as a bolus, electuary or paste.
A tablet may be made by compression or moulding, optionally with one or more accessory ingredients.
Compressed tablets may be prepared by compressing in a suitable machine the active ingredient in a free-flowing form such as a powder or granules, optionally mixed with a binder, lubricant, inert diluent, lubricating, surface active or dispersing agent. Moulded tablets may be made ,. SUBSTITUTE SHEET (RULE 26) by moulding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent.
The tablets may optionally be coated or scored and may be formulated so as to provide slow or controlled release of 5 the active ingredient therein.
Formulations for parenteral administration include aqueous and non-aqueous sterile injection solutions which may contain anti-oxidants, buffers, bacteriostats and 10 solutes which render the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents. The formulations may be presented in unit-dose or multi-dose containers, for 15 example sealed ampoules and vials, and may be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example, saline, water-for-injection, immediately prior to use.
Extemporaneous injection solutions and suspensions may be 20 prepared from sterile powders, granules and tablets of the kind previously described.
Formulations for rectal administration may be presented as a suppository with the usual carriers such as cocoa butter or polyethylene glycol.
Formulations for topical administration in the mouth, for example buccally or sublingually, include lozenges comprising the active ingredient in a flavoured basis such as sucrose and acacia or tragacanth, and pastilles comprising the active ingredient in a basis such as gelatin and glycerin or sucrose and acacia.
Preferred unit dosage formulations are those containing an effective dose, as herein below recited, or an appropriate fraction thereof, of the active ingredient.
SUBSTITUTE SHEET (RULE 26) It should be understood that in addition to the ingredients particularly mentioned above, the formulations of this invention may include other agents conventional in the art having regard to the type of formulation in question, for example those suitable for oral administration may include flavouring agents.
The compounds of the invention may be administered orally or via injection at a dose of from 0.001 to 2500 mg/kg per day. The dose range for adult humans is generally from 0.005 mg to 10 g/day. Tablets or other forms of presentation provided in discrete units may conveniently contain an amount of compound of the invention which is effective at such Hosage or as a multiple of the same, for instance, units containing 5 mg to 500 mg, usually around 10 mg to 200 mg.
The compounds of formula (I) are preferably administered orally or by injection (intravenous or subcutaneous). The precise amount of compound administered to a patient will be the responsibility of the attendant physician. However, the dose employed will depend on a number of factors, including the age and sex of the patient, the precise disorder being treated, and its severity. Also, the route of administration may vary depending on the condition and its severity.
As utilized herein, the term "lower alkyl", alone or in combination, means an acyclic alkyl radical containing from 1 to about 10, preferably from 1 to about 8 carbon atoms and more preferably 1 to about 6 carbon atoms.
Examples of such radicals include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, iso-amyl, hexyl, octyl and the like.
SUBSTITUTE SHEET (RULE 26) The term "lower alkenyl" refers to an unsaturated acyclic hydrocarbon radical in so much as it contains at least one double bond. Such radicals containing from about 2 to about 10 carbon atoms, preferably from about 2 to about 8 carbon atoms and more preferably 2 to about 6 carbon atoms. Examples of suitable alkenyl radicals include propylenyl, buten-1-yl, isobutenyl, pentenylen-1-yl, 2-2-methylbuten-1-yl, 3-methylbuten-1-yl, hexen-1-yl, hepten-1-yl, and octen-1-yl, and the like.
The term "lower alkynyl" refers to an unsaturated acyclic hydrocarbon radical in so much as it contains one or more triple bonds, such radicals containing about 2 to about 10 carbon atoms, preferably having from about 2 to about 8 carbon atoms and more preferably having 2 to about 6 carbon atoms. Examples of suitable alkynyl radicals include ethynyl, propynyl, butyn-1-yl, butyn-2-yl, pentyn-1-yl, pentyn-2-yl, 3-methylbutyn-1-yl, hexyn-1-yl, hexyn-2-yl, hexyn-3-yl, 3,3-dimethylbutyn-1-yl radicals and the like.
The term "heterocyclic radical" means an unsaturated cyclic hydrocarbon radical with 3 to about 6 carbon atoms, wherein 1 to about 4 carbon atoms are replaced by nitrogen, oxygen or sulfur. The ~~heterocyclic radical's may be fused to an aromatic hydrocarbon radical.
Suitable examples include pyrrolyl, pyridinyl, pyrazolyl, triazolyl, pyrimidinyl, pyridazinyl, oxazolyl, thiazolyl, imidazolyl, indolyl, thiophenyl, furanyl, tetrazolyl, 2-pyrrolinyl, 3-pyrrolinyl, pyrrolindinyl, 1,3-dioxolanyl, 2-imidazolinyl, imidazolidinyl, 2-pyrazolinyl, pyrazolidinyl, isoxazolyl, isothiazolyl, 1,2,3-oxadiazolyl, 1,2,3-triazolyl, 1,3,4-thiadiazolyl, 2H-pyranyl, 4H-pyranyl, piperidinyl, 1,4-dioxanyl, morpholinyl, 1,4-dithianyl, thiomorpholinyl, pyrazinyl, piperazinyl, 1,3,5-triazinyl, 1,3,5-trithianyl, SUBSTITUTE SHEET (RULE 26) benzo(b)thiophenyl, benzimidazonyl, quinolinyl, and the like.
The term "aryl" means an aromatic hydrocarbon radical of 6 to about 14 carbon atoms, preferably 6 to about 10 carbon atoms. Examples of suitable aromatic hydrocarbon radicals include phenyl, naphthyl, and the like.
The terms "cycloalkyl" or "cycloalkenyl" means an "alicyclic radical in a ring with 3 to about 10 carbon atoms, and preferably from 3 to about 6 carbon atoms.
Examples of suitable alicyclic radicals include cyclopropyl, cyclopropylenyl, cyclobutyl, cyclopentyl, cyclohexyl, 2-cyclohexen-1-ylenyl, cydlohexenyl and the like.
The term "alkoxy", alone or in combination, means an alkyl ether radical wherein the term alkyl is as defined above and most preferably containing 1 to about 4 carbon atoms. Examples of suitable alkyl ether radicals include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, iso-butoxy, sec-butoxy, tert-butoxy and the like.
The terms "lower alkylene", "lower alkenylenes" and "lower alkynylene" refers to hydrocarbons containing 2 to 10 carbon atoms, preferably 2 to 8 carbon atoms, and more preferably 2 to 6 carbon atoms.
The term "halogen" means fluorine, chlorine, bromine or iodine.
The term "prodrug" refers to a compound that is made more active in vivo.
As used herein, reference to "treatment" of a patient is intended to include prophylaxis.
SUBSTITUTE SHEET (RULE 26) The follo~riag qen~ral synthe~~.c ses~uences are useful in avalrinq the Qresent ~nvent~.on. Abbreviat3,oas used in the schemes ate as foll.a~ws: "Hoc' represents tert-butylogycarbonyl, "z" or ~baz~ represents 10 baazylaxycarbonyl "F'moc° zepresents 9-flnorenyl.meth~ycaxhoayl, "DIBEA" represents diisopropylethylamine, "pllF" represents da.methylicrmaraide, and "TBTtJ" represents 2-(~.8-benza~tfozole-1-yi)-1,1,3,3-tetrametityl uranium 15 tetrafluoroborate.

5chmmve 1 Ra R~H
a ~ Aa' Ra 1~1~ O
O R~ X
R
H2N~-X ARC .~.~.-~- x BocH 1 ~oc c a ~ 2HC1 N~ O
R~ NrX ,pg3 ~~2 1 a. N-Chlorosucc9.ni~nide, DMs h. 820 pa=9.5-10 c. HCl O
~iX ~3 Scheme 2 a,b Ni R R4 I~ O

H2NiX AR3 c RAH Rl R~HN Rl d Y
R4 ~ 2HC1 I~ O
2/ \ -X 3 R NH ~AR
RAH Rl a. acid chloride or chloroformate or isocyanate or sulfonyl chloride or sulfinyl chloride b. hydrogenation c. H20 pH=9.5-10 d. HC1 SUBSTITUTE SHEET (RULE 26) WO 97/32844 PCT/US97l03282 Scheme 3 I~ O

.X
R2 NH ~OEt a N 2 ~Rl R9 l~

RZ NH OOH
N ~Rl ~O

a. R9C02H
TBTU/DIPEA/DMF
b. hydrolysis SUBSTITUTE SHEET (RULE 26) Scheme 4 O
HO~ N~X OCH3 H ~ Ri HN~
z a NH
NH O
R2 N~ 3 .-> R2 N~ X OH
OH I ~ Rl s-C
r NH
R2 ~ N' O
b r R2 N~ X OH
Rl R6 O ~2 O
a. R2C(OEt)=NH
b. HBr/AcOH or catalytic hydrogenation followed by hydrolysis c. acid chloride SUBSTITUTE SHEET (RULE 26) Scheme 5 O
H C' 'N' H3 ~ RI
~x R
a ~r ~"OH
R~
~z R
b,c~d NH
RN~~ H
~ Ri ~z R
a. hydro7.yeis b, CuCO~
c. R~C(4~t)aNH
d. ~owe~'" 50 Scheme 6 O
HO~ N~ X OCH 3 H ~ Rl HN~
Z
i HO~ NH ~ O
HO
R2 N~ X OCH 3 ~ H O

OH HN R b R2 N' X OH
Rl \Z OH ~2 c ~ NH
R2 w N~
O
O

b O
X
R2 N ~ ~OH

~R

a. R2C(C1)=NOH c. acid chloride b. HBr/AcOH or hydrogenation followed by hydrolysis SUBSTITUTE SHEET (RULE 28) Scheme 7 I~O~ N''X H3 H ~,Ri a NH O
l~Ii R~rr'x 3 ~
Ri ~ R~N",X H
~H y ~ R1 ~ ~2 c n o Rr°Y p b ~ii ~z a. RZG(~~t~=NH
b. 8Br/AcOH or hydrogenation ~olaowed by hydrolysis c. acid chloride Or ohlorp~ormate or isocyanate WO 97!32844 PCT/US97/03282 Scheme 8 O
HO~ N~ X OCH 3 H ~ Ri HN~
Z
a HO~
NH
HO~
X IH O
RZ N' OH b R2 N~ X OH
i OH NH R
a c R6 O ~ c. acid chloride w NH
O ~ ,X
R2 ~ N
O
~6 b IH O
O R2 N~X OH
~ Rl p NHZ

a. R2C(C1)=NOH
b. HBr/AcOH or hydrogenation followed by hydrolysis c. acid chloride or chloroformate or isocyanate SUBSTITUTE SHEET (RULE 26) Scheme 9 Q O
H N'~ H3 b .~~ .
6~
a OH

NH R
R
b,C,d p4 a. hxaroiysa.s CrrC03 c. RZC(C1)~NRø
d . ~owex"' S 0 3~1 Scheme 10 H~~~C ~3 Rl Hrl.
Z
O a R6'Y ~ N
R N~ -~ ~~x.:ri ~
A

Rd'Y ~N
X
R N' N
x H ~R
a. R2c~a~,t~=~z~x6 b. ~r/~lc0s or hydrogeXtaticri Zall,owec3 by hydroly~ie Scheme 11 Q
Z~
a,hl0 d d,e,f HO~
RfY
a. formaldehyde b. acid ehlarida or chldroformate or ~.~acyanete a. HZ, Pd/C
d. R~C(Cl)'NOH
a . diethyl amiaa f. hyrdroi~rsis Scheme 12 CI
~I >
N'OH N'OH
Me Me Me-~-Me Me-~-~Me O
O
O b~ ~ HON ~NH
O
HCl ~NH O
Me N
H2N Ow/ H O
O
Me Me--~-Me O
HON O~NH d > HON NH2 ~I O
Me"N W/ Me~NW/
H ~ H
O
a. N-chlorosuccinimide b. chloroacetaldoxime, pH 8.5 c. cation exchange resin d. HC1/EtOH
SUBSTITUTE SHEET (RULE 26) Scheme 13 O~O a O
Me O NH ~ O ~NH
Me~O~ O_ t-Bu0"N~ O
Me/' N r H H
H O O O
O
b Me ~ O~N_H c Me~, Me O H O NH O M
Et~ON~N~OkMe O H
i I
O
O~NH d' a > N.OH O~f~H
II
H2N O NH Me~ N NH Me ~ _ _ H O
Et'ON~ NH2 Et'O~r 1~ ~N~OH
O O H
~ 3HC1 N.OH NH2 Me'~ N NH Me H ~ pN~ N ~N,OH
O H
a. isobutylchloroformate/N-methylmorpholine (NMM)/DMF/dichloromethane (DCM) b. ~-Boc-L-Lys-OEt HCl/NMM/DMF
c. HC1/EtOH
d. chloroacetaldoxime, pH 8.5 e. cation exchange resin f. Pd/C/EtOH/HC1 SUBSTITUTE SHEET (RULE 26) WO 97!32844 PCT/US97I03282 Scheme 14 H
H
O~O a, b N' ~ ~ O>
NH2 ~ ~ I H ~~O ~ M~N~H I-~~O
M~N~
O
O HCl O
Et0 TFA Et0 HO
H ~H
Ilr M~~H NHZ
M
f~ fi --~ --r V~O -O

~H
M~~H ~H2 ~ 3HC1 I _ " ~O
OOH
M
H
O
a. methyl acetimidate/N,N-diisopropylethylamine/DMF
b. reverse phase column chromatography c. hydrolysis, chloride anion exchange column if necessary d. ar-Boc-L-Lys-OEt/1-hydroxybenzotriazole hydrate/DMF/[(N,N-dimethylamino)propyl]ethylcarbodiimide hydrochloride, then reverse phase column chromatography e. 4N HC1/EtOH
f. chloroacetaldoxime, pH 8.5, then cation exchange resin g. Pd/C/EtOH/HC1 SUBSTITUTE SHEET (RULE 26) Scheme 15 M Me wiuo~n M2 m a nr~r.
b HIS
a c a. DL-isopropylideneglycerol/DMF/1-hydroxybenzotriazole hydrate/[(N,N-dimethylamino)propyl]ethylcarbodiimide hydrochloride -b. 5% Pd/C/ethanol/hydrogen c. chloroacetaldoxime, pH 8.5, then cation exchange resin d. HC1/EtOH
SUBSTITUTE SHEET (RULE 26) Scheme 16 NHZ dHBOC munn~
M Me O ~ Me 1~I HI~I~O a > O' b c o IH >
O o nu a TFA
a pIHBOC
nu a. 1,3-benzylideneglycerol/DMF/1-hydroxybenzotriazole hydrate/[(N,N-dimethylamino)propyl]ethylcarbodiimide hydrochloride b. 5~ Pd/C/ethanol/hydrogen 10 c. chloroacetaldoxime, pH 8.5 d. H3B03/(Et0)3B, reverse phase column chromatography e. HC1 SUBSTITUTE SHEET (RULE 26) Scheme 17 ~IHZ ~IHZ
BOC~ a..~ a BOC~ ~ O
N > H NEt2 O
NHZ
N.~ ~IHZ
0~ ~ O
H2 NEt2 M~N NEt2 HCI 0 H HCl O
d OOH 2HC1 .NH2 O
M~N~ v _NEt2 H
O
a. 2-chloro-N,N-diethylacetamide/triethylamine/NaI
b. HCl/HOAC
c. chloroacetaldoxime, pH 8.5, then cation exchange resin d. 5~ Pd/C/ethanol/hydrogen/HC1 ' SUBSTITUTE SHEET (RULE 26) Scheme 18 O NHBoc dH2 H~[ b, c, d -i O ~N
H

NCH HfY
Me' 'N
H
..
a. N-Boc-Y-aminobutyric acid/1-hydroxybenzotriazole hydrate/DMF)/[(N,N- dimethylamino)propyl]-ethylcarbodiimide hydrochloride b. 5~ Pd/C/ethanol/hydrogen c. chloroacetaldoxime, pH 8.5, then cation exchange resin d. HC1/EtOH
SUBSTITUTE SHEET (RULE 26) Scheme 19 N
Hd~O
dH2 a Z\ O b Z~ N O ~ ~ N ~/ --H H
NHBoc NH 2 ~Nj ~ f Hpl H c, d; e~ N,pH 2HC1 Hd Z~N~~~ ~ M~N
H O H O
a. carbonyldiimidazole/imidazole/THF
b. t-butyl N-(2-aminoethyl)carbamate/THF
c. 5~ Pd/C/ethanol/hydrogen d. chloroacetaldoxime, pH 8.5, then cation exchange resin 1Q e. HC1/EtOH
SUBSTITUTE SHEET (RULE 26) Scheme 20 O O
~N'X OH a ~ ~N'X Act H Rl NH-Boc H Rl NH-Boc O O
H2N~X S_R3 ~N~X S_R3 Rl NH-Boc b H Rl NH-Boc c ~N
2"C1 R
R4w N O R4\ .2HY
N O
R2~ N' X S-R3 ~ ( X
d R2~ N~ S-R3 H Rl NH-Boc H
Rl NHZ
Act = amino acid activating group; Y = pharmaceutically acceptable counter ion; a) reagents) required to activate carbonyl to displacement by a nucleophile; b) H2 and catalyst; c) base; d) HC1 in solvent such as dioxane.
SUBSTITUTE SHEET (RULE 26) Scheme 21 O
O
OH - ~ .X
a w Rl NH-Boc H Act RI NH-Boc HO-CH 2Ph o r Z~H~X O-CH2Ph ~' b ' Z~, X
Rl ~ H~ ~O-CH2Ph Rl NH-BoC
c r O O
Z~N~X O-CH2Ph ~s H2N'X OH
H Rl NH~O d Rl NH~O
O O

~N
a ~N
O
.X
R N ~OH
H Rl NH~O
O
Act = amino acid activating group; Y = pharmaceutically 5 acceptable counter ion; a) reagents) required to activate carbonyl to displacement by a nucleophile; b) HC1 in solvent such as dioxane; c) formaldehyde, Cs2C03, acetic anhydride; d) H2 and catalyst.
SUBSTITUTE SHEET (RULE 26) Scheme 22 O O
O-CH2Ph --~. ~N-X O-CHZPh Rl y/° a H Rl NH~O
Ac O
O
b R4w R4w N

I °
RZ N, X OH R Cl X
H HZI~Y ~OH
Rl NH~O ' Rl NH~O
A~ ~ A
O O
Act = amino acid activating group; Y = pharmaceutically acceptable counter ion; a) Cs2C03, acetic anhydride; b) H2 and catalyst.
Scheme 23 R4~ N R4w O ~ O
R2~ ~X O " 2~ ,X
H a R N ~ NH
Rl NHZ H Rl N
H O
a) potassium cyanate, HC1 (aq) [see Bull. Soc. Chim. Fr.
1954, 812, 815].
SUBSTITUTE SHEET (RULE 26) Seh~me 24 NHSoc NHBoc H
a It N ~x H~'s"~x D
D NN
a ~ 2HC1 H Ri R ~S H2 Rt R~ N~ A~ g~ h~l~ A~ 3 X R~ ~;_ X 1~
NH ' H~ p C NH
HBx ~ H zHCI NH2 R
IICl N R tA R2 N
~X 'R O
O N RB
N GIs ~S/
~S/
a) R2G'~~(pEir)'gC~.
b) R~SC~ , c) HCl / EtOx Scheme 25 /OH
N O
R2 ~ ~ 3 R
F

R2 N'~-O
O
R5~ NIX i ~R
R~ Rg a} 0 SUBSTITUTE SHEET (RULE 26) Scheme 26 ~z ~Boc b,c,d r' ~ 2HC1 ~2 a. (Hoc)20 b. RZC(C1)FNR4 C. DOWSX"' S0 d. is HCl Without further elaboration, it is believed that one skilled in the art can, using the preceding descriptions, utilize the present invention to its fullest extent.
Therefore the following preferred specific embodiments 5 are to be construed as merely illustrative and not limitative of the remainder of the disclosure in any way whatsoever. Compounds containing multiple variations of the structural modifications illustrated in the preceding schemes or the following Examples are also contemplated.
All experiments were performed under either dry nitrogen or argon. All solvents and reagents were used without further purification unless otherwise noted. The routine work-up of the reactions involved the addition of the reaction mixture to a mixture of dither neutral, or acidic, or basic aqueous solutions and organic solvent.
The aqueous layer was extracted n times (x) with the indicated organic solvent. The combined organic extracts were washed n times (x) with the indicated aqueous solutions, dried over anhydrous Na2S04, filtered, concentrated in vacuo, and purified as indicated.
Separations by column chromatography were achieved with conditions described by Still. (Still, W. C.; Kahn, M.;
Mitra, A. Rapid Chromatograhic Technique for Preparative Separation with Moderate Resolution. .T. Org. Chem., 1978, 43, 2923-2925.) The hydrochloride salts were made from 1N HC1, HC1 in ethanol (EtOH), 2 N in MeOH, or 6 N HCI in dioxane. Thin layer chromatograms were run on 0.25 mm EM
precoated plates of silica gel 60 F254. High performance liquid chromatograms (HPLC) were obtained from C-8 or C-18 reverse phase columns which were obtained from several vendors. Analytical samples were dried in an Abderhalden apparatus at either 56°C or 78°C. 1H NMR
spectra were obtained from either General Electric QE-300 or Varian VXR 400 MHz spectrometer. 13C NMR spectra were obtained from a Varian spectrometer at 125.8 MHz.
SUBSTITUTE SHEET (RULE 26) a~a ~.
N~-(1-(hydrogyimino)ethyl]~L-lysine, d3.hydrochloxide ~ 2~IC1 NHz H C-" C H
Oz ac.-171) To a 125 mL flask was added 3 c~ ( 4 .812 meal ) of a-~oc~T.-lysi.na and 70m~ of water. This solution was ad~u~sted to p8 ~ 9.5 by addition of 2.5 N Naoe. xo this 10 solution was added pr~rtion wise, 2.3 g of Gh~GIDaCe'~a1db87.~U6 which was prepared immadsately prior to nsa by the reaction of 3.55 g (0.0~ mo1) of acetaldoxima with 10 .4 g ( 4 . 78 nral ) of ~1-chloroauacanimide in b5 aa~ of N,1~-d3.methylformaa~ide at 0 15 °C. The chloroacetaldoxime was isolated after three hours by extracting into da.ethyl ether and washing With aqueous NaCI. Drying with Mgso~, filtration and coaGentration under 3D °G afforded the chloroacetaldoxi~s as a pale yellow oil. During the chlomacetaldoxime addition, the ZO pH Haas kept st 9.5 via concomitant addition of 2.5 N
NaDH. After the addition was compl~te, the solution was allowed to stand at 25 'C for 25 minutes. Thl~ solution was then adjusted to pH = v .5 with 1P xci and poured onto a Dower'" 50 Catioa exchange oolusm. The aoluntn wa8 washed Z5 with water. The Hoc-protected product was then e~.uted with 10% aqueous pyridine.
S3-1) After corleentraW .ng, the product was dapratected by allowing it to stand in 2N HCl at 25 °C
for two hours. concentxatitlg i11 vaGUO afforded 2.9 q 30 (78%) of h-N6-(aximi.naethyl)lysine dihydrochloxide as a viSCaue yellannr Qil. lii-HI~(D~O) 1.25-1.45 (m, 28), 1.5-1.8 (m, 2H), 1.75-1.9 (m, 2x), 2.05 (s, 38), 3.Z5 (t, 28), 3.95 (t, 18); Mass Spectra, H+g~ 2o~.

EXhMFX.B Z
2-amp.na-5-[[1-(hydroxyimino)ethyl.]amz.no]-2_ 5 methylpentanoic acid, dihydrochiorida HO~
~ 2HCt OzH
H v 3 ~
HgC~N~ii oc-Methyl-n,L~ornithi~ne hydroahlaride is protected as a 10 popper eo~sp7.ea via reaction with cugrf.a carboaats in Water at ~eeflnx. This protected amiuo~.ac~.d is then reacted with ah~.aroacataldoxime as in Bxample.l. The pxoduct is eluted from l~owe~r~' with 7.bZ a~ariium hydroxide, Concentration in vacno, fallowad by acidification raith 15 hydrochloric acid affords the title oo~pound.
~ 3 20 1~i6~-[ 1-( hydxoxy~.nino )ethyl ]-2~~~me~thyl-lysine, d~.hydrxhaoride H
~N ~ ZHCI gZ CHs .
HgC' ' CO H
25 83c-3A~ A suspension of Zyeine ethy~.ester dihydrochloride (33 g; 0.14 mole) and Hgs44 (3~ g; 0.28 moles) in a solution of 4-whlora-benzaldehyde (39 g; 0.28 moles) and acatonitril~ (500 mx,) was stirred whale ~1,~1-diisopropylethylamine (36 g; 0.28 moles) was added.irr 30 portions over 1/2 h. xhe mixture was stirred nor 12 h, filtered, concentrated to a s~a3.l. volume, and diluted , with 500 ~t~I. of diethyl ether. The ether s4lution v~ras washed with D.1% aqueous Ha8C0~, aqueous 2 N NaoB
contaia.7.rig 2 g/100 ml of NHZQ8.HC1, again with 0.1%
5 aqueous NaSCO3 and-saturated aqueous Na0l. After drying with Mgsog and removal of the solvent in vacno, ethyl H, N'-..di(4-chloro~pheriylmethylene)-h-lysine was obtained as a vl8ar laguid.
~-3g) The liqua.d was tritnrated with hezanes and the 10 resulting solid was washed with hezanes. This partially purified intermediate was dissolved in 204 mL of THF and stirred is an acetane/dry ice bath. Sodiwn bis-(tximet,hylsilyl)amide in THE' (1I mL, 1 H solution) was added dropwise over 3D min. After ane hour, methyl iod3.de 15 ( 0. 8 g; 13 um~oles ) in THF teas added dtpp~riec~. The reaction mature was slowly warmed up to room temperature and Stirred overnight. the mixture was diluted with water, sad extracted with ethyl ether. The ether estxact was washed with D.1% aqueous Na8CO3 and saturated aqueous ZO NaCl sad conceatrated to yield crude ethyl N.N'-di(4-chlvxa-ghertylmethylene)-a-methyl-D,L-lysine (H + ii= 434).
E~-3C) . This material (4 g) was dissolved in ethyl ether (10O ml) was stirred vigorously with 1 N sCl (50 a4i~) for 2 h, the layer was separated and the a9ueous 25 phase was washed with ethyl ether. The aqueous solutioa was further acidified by the addition of conaentratsd ~iG~.
to 6 D1 and was heated td reflex for 16 h. The aolutaon was cooled to room temperature, and rotary evaporated to dryness. The Residue was dissolved in water and applied 30 to a Dowes:'"~ 50 x 4 (hydxogea form) . mhe colu~pa was washed with warxr, and then 1D% pyridine. a-.t~tethpl-D,Ir-lysine, (3'i + 8 a 161 ) was Cluted from the coluuin With 1 M NH~oB.
8a-3D) Ths dc-methyl-D,l,-lysine 1S protected as a eoppeac cau~lex via reaction with cupri.o aarbc~nate in 35 refluzing water.
E7t-3) Thss protected amino aoid is then reacted with ahloroacetaldoxime as described in Rxample 1. The product is eluted from Dowea"" v~rith 1Di aramoaium hydraxid~.
Concentration in vacuo, fol7.aw~ed by acidification With hydrochloric acid affords the title Compound.

2-L LZ-L L i-(lzYd~Ry~o)ethyl)a~oino) ethylseleno)methyl.]-xTalanir~e, ds.hydrochloride 1o x~N ~ 2I~C~ g z g ~ ''~~~Cp I~
3~ ,~,I z ~-~,p) p,h-Seleriocystiae ( 117 ~tg: 0.5 mmoles, purchased frog Sigma) was snspendad 3..n 15 ml. of nitrogen 15 (N2) gas-pnrged water. sodium boxohydride (38 mg; 1 mole) was added. The reaction mixture beca~one clear in a few minutea_ ~rfter 2 h at room temperature, 2-bromoethylamine HC1 (1.2 g: 6 mmoleS) was added and the reaction mixture was stirred fot 1~ h. The reaction was applied vn to a power" 50 x ~ (hydrogen, forst) aoW ~,. The ooiun~ vrss gashed with. water and 10% ppridine and 2-a~naethyl-selenoeyateine vas eluted faith 1 M NH40H.
E7t-d~s) The 2-amiaoethyl-aelonocysteine is protected and subsequently methylated as described in Example 3 to 25 afford the a-methyl-(2-aminoetriyl)selenocys~eine.
Eu-4C) The a-methyl-(2-aminoethyl) selenocyateine is protected as a copper cot~lea via reactifln with aupr,ic carbonate iri retluaing grater.
~t-4) This protected amino acid is than reacted with 30 chloroaaetaldoxime as de&aribed in Hxample 1. The product is eluted ~zom Dov~fex'" with 1N ammouissm hydroxide.
Cnncentrat.3on in vacua, followed by acidification with hydrochlor~.c acid affordwthe title cor~pound.

~a5 BXA~p~LE 5 ~5.. l 1- ( hydxoaryamino ) ethyl ) -2- ( hydroxymethyl ) -lys i.Re, dil~ydroahlos~ida H
~ 2HCl ~z gZt~~
coZ~
Ex-5A) To an ice-cold starred mi~tture of 1~-~bx L-lyaine (14 g; b.05 moles, purchased from Sigma) in 2.5 Ia 10 Naoe (24 raX,), bepzoyi chloride (10 g) vas added gradually. The pH of the solution eras maintained at 10.5-10.9 by addition of 2 N NaoB. Tho rnix~t~re was stirred at roam~temgeratuxe for 1 h sad filtered. Tha filtrate was eatract~ed with a small amdnat of ethyl acetate arid the 15 organic layer was dried over sodium su7.iate. Th~a solid was removed bx filtration and the filtrate was'evaporated ~ro dryness .
~5B) The crude oia.y N~-Cbz-Na-be7c~zoy7.-lysa.ne residue (6 g) wss heated at 90-100 °C 3.n acetic anhydride (100 20 ml.) for 30 min. Tha aW ttnre was tl~n evaporated. The residue was di.9solved in pyridine and treated with aqueous~fos~maldehyde (35% solution, Fisher). The mixture was starred for 8 hr and then diluted. The reactio~r mixture was kept at 10 °C overnight. T'he preGSp~.tatad 25 erode material mss hydrolyzed by boallnq in 5 N HC1 for 5 h. T'he reavtion mixture was cooled and filtered before being evaporated. The solid residue wag daasoLvBd in water and passed through DosvexT" 50 x .4 (hydxogea tor'm) coiman. 0~--Hydroxyroethyl-D,x~-~ly9ine (!~ + H ~ 17i) was 30 eluted with 1 N NH~QH.
sx-5c) mhe cx--hydroaymethyl-D,z-Lysine as protected as a copper eomplsx via reaetion with cupric carbonate in water at refiux.

gx-5) This protected a~i.no aciQ is then reacted with ohlosoacetalc3oa.ime a~ described in 8xample 1. The product is eiutad from Dower" w].th ~.N ammoaium hydroxide.
Co~ntra~tion in vacuo, followed toy acidification with 5 hydrochloric said affords tha title compound.
s~~ s 10 ~ N6-(1~-(hydraxy~o)-2,2,2-txifluoroethy~.]-D-lysine, dihydrochloarids H~ ~ 2HC1 s F3~~ OZH
15 oc-8oc-L-lysine is reacted with chloxotrifluaroacetaldoxime (,7. Org. Chew. 49, (1984) 919-922) as desara.bed in 8sampl.e 1 to afford the title campoand.

~~ 7 N~-(1-(hydrcxyimino)ethyl]-L-lps~.ri9, ethyl ester, dahydrachlaride H~~~ ~ 2HC1 ~1H2 H C' ' F~
Rf-7A) To a 125 ~G flask is added 0_012 mol of a-BaC-L-lysine ethxl ester hydrxhlarxde and 70 n~ of water.
30 This solution is adjusted to pa = B.5 by addition of 2.5 N Naog. To this solution is added portion wise, 2.3 g of chlaroacetaldoriua which ie prepared immediately prior to nee by the. xeaetion of 3.55 g (O. a6 mbl) of acetaldoxime ~rith 10 .4 g ( 0.98 viol ) of N-chlorasuccinimide in 65 mI. of 5 1~,N-d3methylfornna~.i.de at 0 °C. The ohloroacetaldoxime i.s isolated after three hours by eatraeting into diethyl ether and washing with aqueous paCl. Drying with Mg50,~, filtering and cancentratinc~ under 30 °C affords the' chlox~oacetaldox~ra se a pale yellow oil. paring the 10 ahloroacetaldoaime addition, the pS is kept at 8.5 via conco~aitant addition of 2.5 i~ NaOa. Attar t'he additi.oa .i.s aomple~C.e, the solution is allowed to stand at 25 °c for 25 minutes. The 6G7~ution ie then adiu9ted to p8 =
7.5 with 1N SCl and poured onto a Dowex'~ 50 Catiot~
15 exchange coltma. The column is waaheLi r~rith water. The Doc-prpteated product ~.s then eluted with 10% aqueous pyridine.
Me D
Hp~N ~~~M
zo ~a 8Z-9) Aftex aonceatrating, the product is degrotected by allowing ~.t to stead is 4N aCl in ethanol at 25 °C for twelve hours. Concentrating in vaauo at 30 °C affords L-25 N6-(hydroximi.noethyl)lysa.ne ethyl. eater dihydr'vohlori.de.
NzAIU~.E 8 90 1~~-[1~(hydroxyimir~o)ethxl]-Y,-.lysine, 2,3-dihydxoxypropyl aster, di.hydrochlaXide ~ 2HC1 N,OH NH2 H
Me/ _N O H
H O
EX-8A) ~-Z-a-Boc-L-Lysine is reacted with DL-isopropylideneglycerol (Aldrich Chemical Co., Milwaukee, WI, USA) in DMF in the presence of 1-hydroxybenzotriazole hydrate and [(N,N-dimethylamino)propyl]ethylcarbodiimide hydrochloride as described in 23c to give 8a.
8a EX-8B) The benzyloxycarbonyl (Z) group of 8a is removed with hydrogenolysis using 5~ Pd/C in ethanol and the resulting deprotected amine is reacted with chloroacetaldoxime as described in the preparation of 7a to give 8b.
8b SUBSTITUTE SHEET (RULE 26) EX-8) 8b is treated with 6M HC1 in anhydrous ethanol overnight, evaporated to a solid, and triturated with ether to give the title compound 8.

N6-[1-(hydroxyimino)ethyl]-L-lysine, 2-hydroxy-1-(hydroxymethyl)ethyl ester, dihydrochloride N~OH ~ 2HC1 ~g2 H
O
Me O OH
4, EX-9A) The process of Example 8 is repeated, with 1,3-benzylideneglycerol (H. Hibbert, N.M. Carter, J. Am.
Chem. Soc., 1929, 51, 1601) replacing DL-isopropylideneglycerol.
EX-9B) After the chloroacetaldoxime step, the benzylidene group is removed with triethyl borate/boric acid as described in J. Med. Chem. 1980, ,23, 9-12 (G. Y.
Paris et a1). The reaction product is purified by reverse phase column chromatography.
EX-9) The title compound is procured after HC1/EtOH
treatment as described in Example 8.

N6-[1-(hydroxyimino)ethyl]-L-lysine, 2-(diethylamino)-2-oxoethyl ester, dihydrochloride SUBSTITUTE SHEET (RULE 26) N,OH ~ 2HC1 ~2 O
O
Me H NEt 2 O
EX-10A) E-Boc-a-Z-L-Lysine is reacted with 2-chloro-5 N,N-diethylacetamide (Aldrich Chemical Co.) in the presence of triethylamine and sodium iodide by the method described in Internat. J. Pharmaceutics, 1990, 62, 193-205 (A. H. Kahns and H. Bundgaard) to give 10a.
L~HZ
BOC~ O
NEt2 H

10a EX-10) 10a is treated sequentially with HC1 4M in glacial acetic acid for three hours and then stripped to 15 remove the Boc group, then chloroacetaldoxime as described in Example 7, and then hydrogenolyzed with 5~
Pd/C in ethanol containing excess HC1 and filtered and stripped to give the title compound.

S-ethyl 2S-amino-6-[[1(hydroxyimino)ethyl]amino]hexanethioate, dihydrochloride HO~ . 2HC1 N
SEt H3C H v v O
SUBSTITUTE SHEET (RULE 26) EX-lla) To a flask containing N-a-Boc-N-E-Z-L-lysine in a suitable solvent is added an amino acid activating agent. This transient intermediate is then reacted with ethanethiol to produce the protected thioester.
NH-Boc ~ N S~
H
O
ila EX-11B) The product of EXAMPLE lla dissolved in an appropriate solvent is combined with a hydrogenation catalyst such as palladium on carbon end hydrogen. This reaction is shaken under pressure for an extended period of time in a standard Parr hydrogenation apparatus to remove the Z-function generating the amino product illustrated below.
NH-Boc S~

O
lib EX-11C) The product of EXAMPLE llb dissolved in an appropriate solvent is reacted with the chloroacetaldoxime whose synthesis is described in EXAMPLE 1 to yield the hydroxyimino material illustrated below.
HO~ N NH-Boc S
H C~ N

O
11c SUBSTITUTE SHEET (RULE 26) EX-11) The product of EXAMPLE llc dissolved in an appropriate solvent is reacted with HC1 to remove the Boc-protecting group and provide the title compound.

N2-[(acetyloxy)methyl]-N6-[1-(hydroxyimino)ethyl]-L-lysine, dihydrochloride O
HO~ ~2HC1 '= OH

O
EX-12A) To a flask containing N-a-Boc-N-e-Z-L-lysine in a suitable solvent is added an amino acid activating agent. This transient intermediate is then reacted with benzylalcohol to produce the protected benzylester product illustrated below:
NH-Boc ~N O
H
O
12a EX-128) The product of EXAMPLE 12a dissolved in an appropriate solvent is reacted with HC1 to remove the Boc-protecting group and provide the product illustrated below.
SUBSTITUTE SHEET (RULE 26) Z~ O
O
12b EX-12C) The product of EXAMPLE 12b dissolved in an appropriate solvent is reacted with formaldehyde, Cs2C03, and acetic anhydride to provide the product illustrated below.
O
~O /~ NH /
z~ o N
H
O
l0 12c EX-12D) The product of example 12c is dissolved in an appropriate solvent and combined with a hydrogenation catalyst such as palladium on carbon and hydrogen. This reaction is shaken under pressure for an extended period of time in a standard Parr hydrogenation apparatus to remove the Z-function generating the amino product illustrated below.
O
O~ NH
OH
HZ
O

EX-12) The product of EXAMPLE 12d dissolved in an appropriate solvent is reacted with the chloroacetaldoxime whose synthesis is described in EXAMPLE 1 to yield the hydroxyimino title material.
SUBSTITUTE SHEET (RULE 26) N6-[1-(hydroxyimino)ethyl]-N2-[[[(methylamino)carbonyl]oxy]methyl]-L-lysine, dihydrochloride O
~ 2HC1 HO~

OH
H3C H v v O

N6-[1-(hydroxyimino)ethyl]-N2-[[(methoxycarbonyl)oxy]methyl]-L-lysine, dihydrochloride O
HO~ N ~ 2HC1 HN ~O"OCH
OH
H C~ N
H
O

N6-[1-(hydroxyimino)ethyl]-N2-[[[(methylthio)carbonyl]oxy]methyl]-L-lysine, dihydrochloride SUBSTITUTE SHEET (RULE 26) O
HO~ ~2HC1 /~.

OH

O

5 N6-[1-(hydroxyimino)ethyl]-N2-[[(phenylcarbonyl)oxy]methyl]-L-lysine, dihydrochloride ~ zHCi HO~ N HN~

O
to N2-acetyl-N2-((acetyloxy)methyl]-N6-[1 15 (hydroxyimino)ethyl]-L-lysine, hydrochloride O O
HO~ ~ HCl OH

O
20 EX-17A) The product of EXAMPLE 12b dissolved in an appropriate solvent is reacted with Cs2CO3, and acetic anhydride to provide the product illustrated below.
SUBSTITUTE SHEET (RULE 26) O O
~O~]V~ /
z~ o y N
H
O
17a EX-17B) The product of EXAMPLE 17a dissolved in an appropriate solvent is combined with a hydrogenation catalyst such as palladium on carbon and hydrogen. This reaction is shaken under pressure for an extended period of time in a standard Parr hydrogenation apparatus to remove the Z-function generating the amino product illustrated below.
O O
~O~ N
OH

O
17b EX-17) The product of EXAMPLE 17b dissolved in an appropriate solvent is reacted with the chloroacetaldoxime whose synthesis is described in EXAMPLE 1 to yield the hydroxyimino title material.

N6-[1-(hydroxyi.mino)ethyl]-N2-[(methylthio)carbonyl]-L-lysine, hydrochloride f SUBSTITUTE SHEET (RULE 26) O
~ HC) ~~~3 OH
II3 ~ ~. v ~dp'L~ 19 N2-[(1,1-dim~ethy7.ethoxyycarboayla-N6_~Z_ (hydroxy~mino)ethyll-L-lxeine, hydrochloride HO' ~ HC1 N'I ~T (CH3)g H~. OH
O

To a i25 mT~ flask wag added 3 q (0.012 mat) of a~soc-L-lysix~e and 70m'L. of water. This solution wan adausted to pH ~~ 9.5 by addita.oa of 2.5 N NaoH. To this ealuxzon was added portion w~.se, 2.3 g of ahloroacetaldoaime which wan 15 prepared ianmediately prior to use by the reactiaa of 3.55 g (4.06 mol) of acetaidosime with 1U,4 q (0.78 moi) of ~i-chlprosuacinitaide in 65 m'G of N,h1-dimethy~.formamide at 0 °C. The Ch3oroaCetaldoXime wss isolated after three hours by eztracting into diethyl ether and washing w~.th aqueous 20 Nacl. ~s-~ring with I~gSO4, filtration arid concentration undex 30 °C af~ordad the chloroaceta~.dosime as a pale yellow oil. wring ~Grie ahZoroacetaldosime addition, the pH was kept at 9.5 via conca~itant addition of 2.5 N
NaO~. After the addition wah complete, the solatiori was 25 a~.lowed to stand at ~5 °C for 25 minutes. The solution yras thers adjusted to pH = 7.5 with 1N ae1 and poured onto a Dvwex~°' 50 eatio~ exchange aoluma. The aoluaua wan washed with water. The Boc-protected product was then eluted with 10% aqueous pyridine. 1H-NMR(D20) 1.25 (s, 9H);
1.4-1.65 (m, 6H), 2.05 (s, 3H), 3.22 (t, 2H), 3.75 (m, 1H); Mass Spectra, M + H = 304.

N6-[1-(hydroxyimino)ethyl]-N2-[(methylamino)carbonyl]-L-lysine, hydrochloride O
HO~ ~ HC1 OH

O
EX-20A) Epsilon-Boc-lysine is allowed to react with methyl isocyanate to afford the urea.
O

Boc ~ N OH
H
O
20a EX-20B) Deprotection with HC1 removes the Boc to afford the amine.
. SUBSTITUTE SHEET (RULE 26) WO 97!32844 PCT/US97103282 O

OH
O
20b EX-20) Reaction with acetaldoximinoyl chloride as in example 1 affords the title product.

N6-[1-(hydroxyimino)ethyl]-N2-[N6-(1--{hydroxyimino)ethyl]-L-lysyl]-L-lysine, ethyl ester, trihydrochloride N OH ~ 3HC1 ~ H
Me~N~ NH2 O
N~OH HN
O~
Me O
EX-21A) e-Boc-L-Lys(Z)-OH (3.8 g, 10 mmol) in 25 mL DMF
and 25 mL dichloromethane (DCM) is reacted with isobutyl chloroformate (1.4 mL, (10 mmol) in the presence of NMM
(1.1 mL, 10 mmol). The resulting mixed anhydride is reacted with 10 mmol 8-Boc-L-Lys-OEt HC1 salt suspended in 25 mL DMF containing 10 mmol NMM. After mixing for 16 hr, the reaction mixture is filtered, the residue washed with DCM (25 mL), and the combined filtrate and wash is extracted by 0.5 M KHS04 solution, followed by water SUBSTITUTE SHEET (RULE 26) extraction, and then brine extraction. The organic phase is dried (MgSOq), filtered, and stripped to an oil. The oil is purified by silica gel chromatography if necessary, giving 21a.

O O
Me Me ~ ~NH
Me~~ N NH p M
~ ~ ~ Bile H O ~~N~~ Me O H
21a EX-21B) 21a is treated with 4 M HCl in ethanol 10 overnight at room temperature. The reaction mixture is stripped to a solid, giving 21b.
O
O~NH

Et'O~~ N H

21b EX-21C) Zlb is dissolved in water, and the pH is adjusted to 8.5 with 2.5 N NaOH. This solution~is treated with a fivefold excess of chloroacetaldoxime as described in example 7, through the elution with aqueous pyridine. The product is stripped to an oil, giving 21c.
SUBSTITUTE SHEET (RULE 26) HO
O~NH
OH
Me H O NH
Et~ tr~~ N Me O H
21c EX-21) 21c is hydrogenolyzed (H2 10 psi) with 5% Pd/C
in ethanol containing an excess of HC1. The title compound 21 is isolated by stripping the solution to a solid, and triturating this solid with ether.

N6-[1-(hydroxyimino)ethyl]-N2-[N6-[1-(hydroxyimino)ethyl]-L-lysyl]-L-lysine, trihydrochloride HON . 3HC1 ~I H
Me~N~ NH2 O
HON HN
O
Me N ~H
H
O
The procedure of Example 21 is repeated, with the HC1 salt of ~-Boc-L-Lys benzyl ester (E-Boc-L-Lys-OBz HCl) replacing the ~-Boc-L-Lys-OEt HC1 salt. The final hydrogenolysis then gives the title compound.
SUBSTITUTE SHEET (RULE 26) N6-[1-(hydroxyimino)ethyl]-N2-[N6-(1-iminoethyl)-L-lysyl]-L-lysine, ethyl ester, trihydrochloride ~H
~I
Me~N~H NH2 3HCl O
N~OH HN
O
Me O
EX-23A) a-Z-L-Lysine ethyl ester in 10 mL DMF is treated with methyl acetimidate (0.692 g, 6 mmol) and N,N-diisopropylethylamine (1.05 mL, 6 mmol) overnight.
Solvent is removed in vacuo and the residue is purified by reverse phase column chromatography, giving 23a.
23a EX-23B) The product N6-[1-iminoethyl]-L-Lys(Z)-OEt trifluoroacetate (TFA) salt (23a) is treated with refluxing aqueous 6M HC1 overnight and stripped to give N6-[1-iminoethyl]-L-Lys(Z)-OH HC1 salt (BP4b). Passage through an anion exchange column in the chloride form can be used if necessary to fully replace the TFA.
SUBSTITUTE SHEET (RULE 26) ~N-H HL~J~O
HCl O
HO
23b EX-23C) To a stirring solution of N6-[1-iminoethyl]-L-Lys(Z)-OH HC1 salt (23b, 13.8 mmol), e-Boc-L-Lys-OEt (39.5 mmol), and 1-hydroxybenzotriazole hydrate (2 g, 14.5 mmol) ~
75 mL of dimethylformamide (DMF) cooled in an ice bath is added [(N,N-dimethylamino)propyl]ethylcarbodiimide hydrochloride (2.8 g, 14.5 mmol). After stirring 55 h at 4_ ambient temperature, the reaction mixture is concentrated it vacuum. The resulting material is dissolved in aqueous acetonitrile and passed through a reverse phase chromatographic column, giving a-(Z)-E-N-iminoethyl-L-Lysyl-Boc-L-Lysine ethyl ester TFA salt (23c).
~H
I
TFA M~ N~H NHZ
O
O H1y O~
O
23c EX-23) This material is treated with ethanolic HC1 as described for 21b, dissolved in water and treated with chloroacetaldoxime, followed by hydrogenolysis, both as described in Example 21, to give the title compound 23.
SUBSTITUTE SHEET (RULE 26) N6-[1-(hydroxyimino)ethyl]-N2-[N6-(1-iminoethyl)-L-lysyl]-L-lysine, trihydrochloride ~H
Me~ NCH NH2 ~ 3HC1 O
N~OH HN
O~
Me H ~ H
O
The procedure of Example 23 is carried out, with E-Boc-L-Lys-OBz replacing E-Boc-L-Lys-OEt, to give the title compound.

N2-(N-acetyl-L-methionyl)-N6-[1-(hydroxyimino)ethyl]-L-lysine, ethyl ester, hydrochloride O
Me' \NH
~ HCl Mew S O
N~OH HN
O
Me N
H
O
The procedure of Example 21 is run through the elution with aqueous pyridine, with N-acetyl-L-methionine replacing e-Boc-L-Lys(Z)-OH. The aqueous pyridine SUBSTITUTE SHEET (RULE 26) solution is stripped to a solid, dissolved in dilute HC1, and shelled and lyophilized to give title compound.

N2-(N-acetylmethionyl)-N6-[1-(hydroxyimino)ethyl]-L-lysine, hydrochloride O
~HC1 Me~NH
Me~S O
N~OH HN
O~
Me N H
H
O
l0 The title compound of Example 25 is treated with refluxing aqueous 6N HCl for 12 hr and then stripped to a solid to give the title compound.

N2-(L-alanyl)-N6-[1-(hydroxyimino)ethyl]-L-lysine, dihydrochloride O
HO~
N

EX-27A) The product of example 1 is dissolved in ethanol, cooled down in ice bath and HC1 gas is bubbled into the solution . This solution is stirred at room temperature followed by removal of the solvent in vacuo to obtain the ethyl ester.
SUBSTITUTE SHEET (RULE 26) EX-27B) This ester is coupled with Na-Boc-L-alanine in dimethylformamide (DMF) in the presence of [2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate] (TBTU) and diisopropylethylamine (DIPEA). The solvent is removed in vacuo and the product is isolated on preparative HPLC using acetonitrile/H20 gradient.
EX-27) Protecting groups are removed in 2N HC1 at reflux. The title compound is isolated on preparative HPLC using acetonitrile/H20 gradient.

N6-[1-(((methoxycarbonyl)oxy]imino]ethyl]-L-lysine, dihydrochloride ~ 2HC1 w N NH2 o i off O

N6-[1-[[[(methylamino)carbonyl]oxy]imino]ethyl]-L-lysine, dihydrochloride ~O~N ~ 2HC1 'I 2 O I OH
H3 H v v O
. SUBSTITUTE SHEET (RULE 26) ~CA~PI~ 30 ~6-fi-III(~tnylthio]carbopyl]oxy)imsrio]ethyl]-1~-lysine, dihydrochloride H3 0~ ~ ZHC~
N ~HZ
~ ~H
H3C'' r:~auryr.n! $1 N6-[1-[(acetyloay)imina)ethyl)-hlysi~e, dihydroch7.oride ~ 2HC1 ~~ N L~1H2 ~~ iDH
U
15 sz-3la) 2ao rr-alpha~boc~L-lyss.ne (Aldrich) in grater at p8 9, is added 0-acetyl-2-chlo~oacetaldehyde ox~na, ad~ust~.ag to p8 9 ~hxaughQUt the zeact~.an with 2.5N
sodium hydxo~ide. Contents are purified on a nowex'"-SO
catian exchange resin, elut~.ng the bnc-protocted product 20 with 1N ammonia hydxox3de.
~--31) Acidic hydrolysis axtd pnri.fication by C18 roverse phase chromatography affoxds the de9ired pXoduct.

N6-f 1~ I I (P~uYlcarbony~. )oxy)~.mir~o]ethyl ]-.L-lysi~, di.hydrochlorid~a WO 97!32844 PCT/US97/03282 Ph O\ N ~ 2HC1 Z
O OH

O

N6-[1-[(methoxyimino)ethyl]-L-lysine, dihydrochloride ~ 2HC1 H3C0~ N NH2 OH

O
EX-33A) To N-alpha-boc-L-lysine (Aldrich) in water at pH 9, is added O-methyl-2-chloroacetaldehyde oxime, adjusting to pH 9 throughout the reaction with 2.5N
sodium hydroxide. Contents are purified on a Dowex-50 cation exchange resin, eluting the boc-protected product with 1N ammonium hydroxide.
EX-33) Acidic hydrolysis and purification by C18 reverse phase chromatography affords the desired product.

N6-[1-[(phenoxyimino)ethyl]-L-lysine, dihydrochloride PhO~ N ~ 2HC1 NH
OH

O
SUBSTITUTE SHEET (RULE 26) N6-hydroxy-N6-[1-(hydroxyimino)ethyl]-L-lysine, dihydrochloride HON ~ 2HC1 OH
H3 N v v OH O

N6-[1-(hydroxyimino)ethyl]-N6-[[(methoxycarbonyl)oxy)methyl]-L-lysine, dihydrochloride HO~ N ~ 2HC1 OH

O
O
H CCY 'O

N6-[1-(hydroxyimino)ethyl]-N6-[[[(methylamino)carbonyl)oxy)methyl)-L-lysine, dihydrochloride SUBSTITUTE SHEET (RULE 26) HO~ N . 2HC1 ]VH

OH
H3C N ~ v O
O

N6-[1-(hydroxyimino)ethyl]-N6-[[[(methylthio)carbonyl]oxy]methyl]-L-lysine, dihydrochloride HON .2HC1 NHZ
OH

O
O
H3CS~0 to 15 N6-[1-(hydroxyimino)ethyl]-N6-[[(phenylcarbonyl)oxy]methyl]-L-lysine, dihydrochloride HO~ N ~ 2HC1 OH

O
O
Ph/ 'O
SUBSTITUTE SHEET (RULE 26) N6-(acetyloxy)-N6-(1-iminoethyl)-L-lysine, dihydrochloride NH ~ 2HC1 NH

OH

OAc O
EX-40A) a-Cbz-protected hydroxylysine methyl ester is prepared as described in J. Org. Chem. 59, 4858-4861 (1994). This material is then allowed to react with ethyl acetimidate to afford the hydroxamidine.
OH

NH HN~
Z
40a EX-40B) Reaction with acetic anhydride affords the acetylhydroxamidine.
O
O

NH HN
~cbz 40b EX-40) Deprotection with HBr in acetic acid affords the title compound.
SUBSTITUTE SHEET (RULE 26) S~uIPL~." 41 r76-t1-f(phenylthio)imino]ethyl-~,-7.ysine, dihydrochloride PhS~N ~ 2HC1 OH
~ ~a ~o N~- ( X~imi.nosthy~l ~ -r16- ( PhenY7.~2tbo~r ) .~L-Lysine, dihydrochiori.de ~ 21~C1 ~z~
m gt..q,~) pmega benzylQxy acetyllysine methyl ester ie prepared as described iri 3'et.~Tret. 25(9), 927-.93~. (1994).
~hxs material is hydrolyzed to afford benzylaxylysine.
Ph'~O~ d2I~
2 Q ~2 42a ~:a-42 ) prvtect~.on of the amino acid with ~ Cu2+ allows reaction with ethy3, aaetimidaxe 3h baei.c water.
25 Purification via Dola~~T~ 50 ion exchange resin affords the title compotind_ WO 97/32844 PCT/IJS97l03282 N6-hydroxy-N6-(1-iminoethyl)-L-lysine, dihydrochloride NH ' 2HC1 NH

OH

OH O

N6-acetyloxy-N6-[1-[(acetyloxy)imino~ethyl]-L-lysine, dihydrochloride AcO~ ~ 2HC1 OH

OAc O
EX-44A) a-Cbz-protected hydroxylysine methyl ester is prepared as described in J. Org. Chem. 59, 4858-4861 (1994). This material is then allowed to react with acetaldoximinoyl chloride to afford the N, N' dihydroxyamidine.
OH

N HN
HO~ ~Z
44a EX-44B) Reaction with acetic anhydride affords the bis-acetoxyamidine.
SUBSTITUTE SHEET (RULE 26) O

O
N ~ HN
~O ~Z

44b EX-44) Deprotection with HBr in acetic acid affords the title compound.

N6-[1-[[(methoxycarbonyl)oxy]imino]ethyl]-N6-[(methoxycarbonyl)oxy]-L-lysine, dihydrochloride H3c o~ ~2HCi I
O ~ OH

O

N6-[1-[[[(methylamino)carbonyl]oxyJimino]ethyl]-N6-[[(methylamino)carbonyl]oxy]-L-lysine, dihydrochloride SUBSTITUTE SHEET (RULE 26) H3CH O~ ~ 2HC1 O ~ OH

O

N6-[1-[(((methylthio)carbonyl]oxy]imino]ethyl]-N6-[[(methylthio)carbonyl]oxy]-L-lysine, dihydrochloride ~ 2HC1 ~_ H3CS O~

O ~ OH

O
to N6-[1-[[(phenylcarbonyl)oxy]imino]ethyl]-N6-15 [(phenylcarbonyl)oxy]-L-lysine, dihydrochloride Ph O ~ 2HC1 ~ N NH2 O OH
H
Ph O
SUBSTITUTE SHEET (RULE 2fi) EuA~~PLB 49 P6-[ 1- ( phenoxyimino ) ethyl -N6.-phenoxy-L-lzrsine, 5 di~ydrochlox3de imo~i ~ zHC~
off C' 'a~
N6- [ 1- imetuoxyimine ) et3ryl ] -.N6- ( phenyhw~y 1-~"lYsiue, dihydxochlor3de H3CO~N ~ 2HC1 ~ OH
H~~ N
I
QCH ZPh. O

sx-50A) One=ga beri2ylosy acetyhysine methyl ester is g~Cepared. as descx~.hed in Tet. yet. 25(9), 927-930 (1994).
This material 3.s hydrolyzed to afford hanzyloxylysixte.
CozH

50a SX-50) P~ateet~.C~n of the amino acid with Cu2t al7.ows 25 reaeta.on with o-methyl acetoxim~.noyl chloride in basic water. Purification via nowex"" 50 ioa exchange resi,a a~fards the title compound.

N6-[1-[[(acetyloxy)methyl]imino]ethyl]-N6-hydroxy-L-lysine, dihydrochloride ~ 2HC1 Ac0~ N NH2 OH

OH O
EX-51A) a-Cbz-protected hydroxylysine methyl ester is prepared as described in J. Org. Chem. 59, 4858-4861 (1994). This material is then allowed to react with CH3C(OEt)=NCH2oAc to afford the hydroxamidine.
OH

Ac~O~ N HN~Z
51a EX-51) Deprotection with HBr in acetic acid affords the title compound.

N6-hydroxy-N6-[1-[[[(methoxycarbonyl)oxy]methyl]imino]ethyl]-L-lysine, dihydrochloride O
~ 2HC1 H3C O~ N NH2 OH

OH O
SUBSTITUTE SHEET (RULE 26) N6-hydroxy-N6-[1-[[[[(methylamino)carbonyl]oxy]methyl]imino]ethyl]-L-lysine, dihydrochloride O
~ 2HC1 H3CH O~ N NH2 OH

OH O

N6-hydroxy-N6-[1-[[[[(methylthio)carbonyl]oxy]methyl]imino]ethyl]-L-lysine, dihydrochloride O
~ 2HC1 H3CS O~ N NH2 OH

OH O

N6-hydroxy-N6-[1-[[[(phenylcarbonyl)oxy]methyl]imino]ethyl]-L-lysine, dihydrochloride O
~ 2HC1 Ph O~ N NH 2 OH
H3C N~
OH O
SUBSTITUTE SHEET (RULE 26) N6-[(acetyloxy)methyl]-N6-[1-(hydroxyimino)ethyl]-L-lysine, dihydrochloride HON ~ 2HC1 NH

OH

O
OAc EX-56A) a-Fmoc, E-Cbz-lysine is allowed to react with formaldehyde followed by acetic anhydride.
Ac~O

Z
NH-fmoc 56a EX-56B) Deprotection of the Cbz via catalytic hydrogenation affords the acetoxymethyl lysine.
Ac~O

NH-fmoc 56b EX-56) Reaction of this material with acetaldoximinoyl chloride followed by Fmoc deprotection affords the title compound.
SUBSTITUTE SHEET (RULE 26) N6-(1-iminoethyl)-N6-[(methoxycarbonyl)oxy]-L-lysine, dihydrochloride ~ 2HC1 OH

O

N6-(1-iminoethyl)-N6-[[(methylamino)carbonyl]oxy]-L-lysine, dihydrochloride ~ 2HC1 NHZ
OH
H3 N v v O

N6-(1-iminoethyl)-N6-[[(methylthio)carbonyl]oxy]-L-lysine, dihydrochloride NH ~ 2HC1 NH

OH

I

O
SUBSTITUTE SHEET (RULE 26) N6-(1-iminoethyl)-N6-[(phenylcarbonyl)oxy]-L-lysine, dihydrochloride ~ 2HC1 NH NHZ
OH

Ph O

N6-(1-iminoethyl)-N6-phenoxy-L-lysine, dihydrochloride ~ 2HC1 OH

OPh O

N6-[1-[(methylthio)imino]ethyl]-L-lysine, dihydrochloride H3CS~ ~ 2HC1 ~2 OH
H3C H v v O
EX-62A) To a flask is added N-a-Boc-L-lysine methyl ester hydrochloride and water. This solution is adjusted to pH = 8.5 by addition of 2.5 N NaOH. To this solution is added portion wise, methylacetimidate hydochloride.
During the methylacetimidate hydochloride addition, the SUBSTITUTE SHEET (RULE 26) pH is kept at 8.5 to lif via concomitant addition o~ Z.5 N
Ha4&. Afte~c the addition is compl~e, the solution is allowed to stand at Z5 °C for 25 minutes. The solution is then adjusted to pH ~ 9.5 with 1N &C1 and poured onto 5 ' a Dowex'" 50 Cax~.oa exahaage oolu~nn. Tha ooluma is washed with water- The Hoc-~pxotected prodact is then elated with 10$ Bqueone pyridine.
Ex-62H) TQ a flask is added the amidine free base in 10 chloznform at -70'c fallo~ed by tha addition of ~than~esulfenylchlaride. The solvent is aoaceatrated under vacuun~arid the resulting material is triturated with ether. The resulting product is oo~.lected by filtration and recrys~tallized fxaaa aloohol.

~a-G2) The product is then deptotected by allowa.ng it to stapd in 2r1 8C1 iri ethanol at 25 °C. The pradact ~.e aLaolated by reverse phase H~C chromatography.
24 EIi~P~ 63 N6-(1-iminoethyl)-N6-(msthylthn.o)-L-lysine, dihydfochloride ~ 2HC1 ,l ~2 O~I
-I
25 ~H3 EX-6311) To a flask is added N-a-aoe-D-lysine methyl ester hydroahlaride ana water. Thin solution is adjusters t0 ps = 8.5 by addi.tiou of 2.5 N NaOH. TP this solution 30 is added pat~tion wise, methylacetimidate hydrochloride.
l7uring the mathylacetiud.date hydrochior~.de addition, the pa 3a kept at 8.5 to 1Q via concomitant addition of 2.5 N
Naoa. After the addition is complete, the solution is al7.owed to stand at 25 °C fox 23 minutes . The solution is then adjusted to pa ~ 7.5 with 1N aCl aat~ gouxed onto a bowex'" 5o Canon axahange aolumu. The column is washed with u~atar. The Hoc-protected pxoduct is then eluted w:~tp 5 10% aqueous pyridine.
E~~.63a) To a flask is added the amiaine free i~aae ~.n chlo~xnfvrm at -~o°c followed by the addition of ~thaneaulfenylchloride. The solvent is conceatxated to nndax vacuum and the resulting material is triturated with other. The resulting product is collected by filtration and recxystal.lized from alcohol.
gx~6~) the product is then deprotected by allowing it 15 to stand in 2N aCl in ethanol at 25 aC. The product is isolated by reverse phase PLC chromatography.
15SAlihLE 6~
2D N6-(1-3~i~loetnyl)-N6-(phenylthio)-L-lysine, d~.hydrochloride ~ 2HC1 ~z ~ OH
H3C"
SPh ~5 Ex111dPhE 65 N5-[1--[tph~Ylthiv)amino]ethyl)~Z-1y83.ne, dihydroGhlorida Ph-SAN ~ 2HC1 ~z QH

5-[4-[[1-(hydroxyimino)ethyl]amino]butyl]imidazolidine-2,4-dione, dihydrochloride HO~ N ~ 2HC1 O
HN--NH
H3C H v v O
The title product of EXAMPLE 1 dissolved in water is reacted potassium cyanate in the presence of HC1 as described in Bull. Soc. Chim. Fr. 1954, 812, 815 to provide the title material N-[N6-[1-(hydroxyimino)ethyl]-L-lysyl]-L-alanine, dihydrochloride HO~ N ~ 2HC]

2S-amino-6-[[1-(hydroxyimino)ethyl]amino]-4-hexenoic acid, dihydrochloride HO~ ~ 2HC1 ~2 OH
H3 H ~ v SUBSTITUTE SHEET (RULE 26) N-[2S-amino-2-[3-[[[1-(hydroxyimino)ethyl]amino]methyl]cyclopentyl]acetyl]-L-5 alanine, dihydrochloride HON ~ 2HC1 O COZH

N-[2S-amino-2-[3-[[[1-(hydroxyimino)ethyl]amino]methyl]isoxazol-5-yl]acetyl]-L-alanine, dihydrochloride HO~ N ~ 2HC1 NH

]~~ O C02H

N2-(4-amino-1-oxobutyl)-N6-[1-(hydroxyimino)ethyl]-L-lysine, ethyl ester, dihydrochloride ~2 ~ 2HC1 O
N~OH HST
OEt Me N
H
O
SUBSTITUTE SHEET (RULE 26) EX-71A) To a stirring solution of N-Boc-y-aminobutyric acid (Sigma Chemical Co., St. Louis, M0, USA), e-Z-L-Lys-oEt, and 1-hydroxybenzotriazole hydrate in (DMF) cooled in an ice bath is added [(N,N-dimethylamino)propyl]ethylcarbodiimide hydrochloride as described in Example 23c. After stirring 55 h at ambient temperature, the reaction mixture is concentrated in vacuum to a semisolid, and partitioned between ethyl acetate and water. The organic phase is washed with water and then brine, dried (MgS04), filtered and stripped to give 71a.
NHBOC

Z~N~ O

7 la EX-71) 71a is treated sequentially with 5~ Pd/C in ethanol and hydrogen at 10 psi, then chloroacetaldoxime as described in Example 7, and then HC1 4M in ethanol for thirteen hours and then stripped and triturated with ether to give the title compound.

N2-[[(2-aminoethyl)amino]carbonyl]-N6-[1-(hydroxyimino)ethyl]-L-lysine, ethyl ester, dihydrochloride ~ 2HC1 NH2 N,OH gL~J
Me' _N Et H O
SUBSTITUTE SHEET (RULE 26) EX-72A) E-Z-L-Lys-OEt is treated with stochiometric amounts of carbonyldiimidazole and imidazole, both from Aldrich Chemical Co., in THF, to give 72a.
~n HL~~O
Z,~~O~
O
72a EX-72B) After 12 hours at room temperature the product 72a is treated with an excess of t-butyl N-(2-aminoethyl)carbamate (N-Boc ethylene diamine, from Aldrich Chemical Co.) to give 72b.
NH-BOC
72b EX-72) 72b is treated sequentially with 5~ Pd/C in ethanol and hydrogen at 10 psi, then chloroacetaldoxime as described in Example 7, and then HC1 4M in ethanol for thirteen hours and then stripped and triturated with ether to give the title compound N2-[(2-aminoethoxy)carbonyl]-N6-[1-(hydroxyimino)ethyl]-L-lysine, ethyl ester, dihydrochloride SUBSTITUTE SHEET (RULE 26) NHZ
~ 2 HCl NJ~H HIy Et Me N
H O
The process described in Example 72 is repeated, except that N-Boc-glycinol (Aldrich Chemical Co.) replaces t-butyl N-(2-aminoethyl)carbamate.

9S-amino-5,6,7,8,9,10-hexahydro-3-methyl-4H-1,2,4-oxadiazecin-10-one, dihydrochloride ~ 2HC1 . ~;H2 The product of example 7 is heated in a high boiling solvent to effect ethanol evolution. Removal of the solvent in vacuo followed by chromatographic purification affords the title compound.

3S-aminohexahydro-1-[1-(hydroxyimino)ethyl]-2H-azepin-2-one, dihydrochloride SUBSTITUTE SHEET (RULE 26) OH

N
~ 2HC1 EX-75A) 3-aminocaprolactam is protected with Boc anhydride to give the bis-protected aminocaprolactam after chromatographic isolation.
Boc N
BoC
75a EX-75B) Formation of the amide anion with a lithium amide base followed by reaction with acetaldoximinoyl chloride affords the hydroxamidine.
OH

N
Boc ~N
Boc 7 5b EX-75) Deprotection with HC1 affords the title product.
SUBSTITUTE SHEET (RULE 26) BioloqiCal Data 5 The subject rompounds of formula (I) have been ar are expected to he found to inhibit nitric oxide synthase sad gasses useful pharmacological properties as demonstrated in one or mole of the following assayss 10 Citrullina Assayr for Nitric g~'de 5vnthase Nos activity was measured by Monitoring the conversion of L-[2,3-3H]-arginine to L-[2,3-aH]_ eitruZline. Mouse indurible t~lo5 (miNOS) was prepared from 15 an extract of ips-treated mouse 3~w,26~.7 cells and rat brain constitutive z~OS (rnN08) was prepared frog an extract of r8t cerebei.lnm. Moth preparations were partially purified by n87~E-Sephaarose'" Chrosatographx.
Sn~yme ( 10 p. L) was added to 40 ~1 h of 50 mM Tris ( pa 7 .6 ) 20 arxd the xeaetian initiated by the add3tioa of 50 [1z of a solution containing 50 ~nM Tris (pg 7 . 6 ) , 2 . 0 mg/ad. bovine serum albumin, 2.0 mM D~1"x, 4.0 mM caClz, 20 p!S ~'AD, 100 &M tetrahydrobiopterin, Z.0 mM LzADPH sad 60 w M h-arginine containing 0.9 ~lCi of 7~-[2,3-3Fi)-argiriine.
25 For constitutive N06, calmodulin was included at a final conrentratioa of 40 nli. Folloraiag incubation at 37 °C
for i5 minutes, the reaction was terminated by addition of 300 ~ L cold buffer aontaini,r~g 10 mM EGTA, 100 mM
HEpES (p8 5.5) and 1.0 mM L-citruls~.ne. The 30 citrulline was separated by chromatography on Dotaes'" Sow X~8 ration exchange resin arid radiaaotf,vity c)uantified with a liquid scintillation counter.
Raw Ceh Nitrite Assay lt~W 26.7 cells are plated to confluency on a 96-well tissue culture plate grown overnight (Z7h) 3.n the presence of LPS to induce NOS. A row of 3-6 wells were left untreated and served as controls for subtraction of nonspecific background. The media was removed from each well and the cells are washed twice with Kreb-Ringers-Hepes (25mM, pH 7.4) with 2 mg/ml glucose. The cells are then placed on ice and incubated with 50mL of buffer containing L-arginine (30mM) +/- inhibitors for 1h. The assay is initiated by warming the plate to 37~C in a water bath for 1h. Production of nitrite by intracellular iNOS is linear with time. To terminate the cellular assay, the plate of cells is placed on ice and the nitrite-containing buffer removed and analyzed for nitrite using a previously published fluorescent determination for nitrite. T. P. Misko et al, Analytical Biochemistry, 214, 11-16 (1993). AlIW alues are the average of triplicate wells and are compared to a background-subtracted induced set of cells (100 value).
In Vivo Assay Rats were treated with an intraperitoneal injection of lOmg/kg of endotoxin (LPS) with or without oral administration of the nitric oxide synthase inhibitors.
Plasma nitrites were measured 5 hours post-treatment.
The results show that the administration of the nitric oxide synthase inhibitor decreases the rise in plasma nitrites, a reliable indicator of the production of nitric oxide, induced be endotoxin.

- SUBSTITUTE SHEET (RULE 2fi) TABLE I
Rodent {Cell Data and in vitro Enzyme Data) Compound miNOS* rnNOS* Raw Cell*
IC50 [~t.M] IC50 [~.1.M]
Example 1 77 1470 28 * miNOS refers to mouse inducible NOS
rnNOS refers to rat brain constitutive NOS
Raw Cell refers to cultured RAW 264.7 cells TABLE II
Human (in vitro Enzyme Data) Compound hiNOS* hecNOS* hncNOS*
IC50 [E.tM]
Example 1 154 1474 907 * hiNOS refers to recombinant human inducible NOS
hecNOS refers to recombinant human endothelial constitutive NOS
hncNOS refers to recombinant human neuronal constitutive NOS
. SUBSTITUTE SHEET (RULE 26) TABLE III
Low Dose LPS*
Compound in vivo Effective Dose (p. o., mg/kg/day) 0.1 1 10 Example 1 0~ inh. 54~ inh. 97o inh.
* Low Dose LPS refers to the in vivo low-endotoxin assay carried out on rats as described above.
From the foregoing description,~-one skilled in the art can easily ascertain the essential characteristics of this invention, and without departing from the spirit and scope thereof, can make various changes and modifications of the invention to adapt it to various usage's and conditions.
SUBSTITUTE SHEET (RULE 26)

Claims (10)

WHAT IS CLAIMED IS:

1. A compound having the formula:
and pharmaceutically acceptable salts, wherein:
A is selected from O or S and may be taken together with R4 to form a heterocyclic ring of 3 to 6 carbon atoms; or A is N when R3 and R7 are taken together to form a heterocyclic ring of 3 to 6 carbon atoms; or R5 and A-R3 are taken together to form a covalent bond; or A is N provided R3 is not a heterocyclic radical of 3 to 6 carbon atoms;
R1 is not present ar is selected from the group consisting of hydrogen, hydroxyalkyls of 1 to 10 carbon atoms, alkoxyalkyls of 1 to 10 carbon atoms, lower alkyls of 1 to carbon atoms and haloalkyls of 1 to 20 carbon atoms;
R2 is selected from the group consisting of straight and branched lower alkyls of 1 to 10 carbon atoms, lower alkenyls of 2 to 10 carbon atoms, and lower alkynyls of 2 to 10 carbon atoms, cycloalkyls of 3 to 10 carbon atoms, cycloalkenyls of 3 to 10 carbon atoms, and haloalkyls of 1 to 10 carbon atoms;

R3 is selected from the group consisting of aryl of 6 to 14 carbon atoms, heteroaryl of 6 to 14 carbon atoms, alkylaryl of 6 to 14 carbon atoms, alkylheteroaryl of 6 to 14 carbon atoms, all optionally substituted by one or more of halogen, nitrile, carboxy, carboxyalkyl of 1 to 10 carbon atoms, carboxyalkylaryl of 6 to 14 carbon atoms; or R3 is selected from the group consisting of H, alkyl of 1 to carbon atoms, alkenyl of 2 to 10 carbon atoms, CH2OC(=O)YR6, alkylhydroxy of 1 to 10 carbon atoms, alkylpolyhydroxy of 1 to 10 carbon atoms, alkyl(poly)oxyacyl of 1 to 10 carbon atoms, alkylcarboxy of 2 to 10 carbon atoms, optionally substituted by one ar more of alkyl of 1 to 10 carbon atoms, hydroxy, amino, carboxy, carboxyalkyl of 1 to 10 carbon atoms, alkylcarbonyl of 1 to 20 carbon atoms;
R4 is selected from H, OH, SH, OR6, SR6, OC(=O)R6, SC(=O)R6, CH2OC(=O)YR6, OC(=O)YR6, SC(=O)YR6;
Y is independently selected from O, S, CH2, CHR6, C(R6)2, NH, NR6;
R5 is selected from H, OH, SH, OR6, SR6, OC(=O)R6, SC(=O)R6, CH2OC(=O)YR6, 4C(=O)YR6, SC(=O)YR6;
provided that R4 and R5 are not simultaneously hydrogen or CH2OC(=O)YR6;
R6 is selected from hydrogen, alkyl of 1 to 10 carbon atoms, alkenyl of 2 to 10 carbon atoms, alkynyl of 2 to 10 carbon atoms, cycloalkyl of 3 to 10 carbon atoms, heterocyclic radical of 3 to 6 carbon atoms, aryl of 6 to 14 carbon atoms, heteroaryl of 6 to 14 carbon atoms all optionally substituted by one or more alkyl of 1 to 10 carbon atoms, alkoxy of 1 to 10 carbon atoms, hydroxy, halogen, trifluoromethyl, nitro, cyano, or amino groups;

R7 is selected from H, S(O)R9, SO2R9, CH2OC(O)-R9, C(O)-R9 where C(O)-R9 can represent natural and synthetic amino acids or R9 can be defined as below, or R7 and R3 taken together comprise a 5- or 6-membered heterocyclic ring containing two or more heteroatoms, optionally substituted with alkyl of 1 to 10 carbon atoms and/or oxygen functions, or taken together comprise a metal complex containing a divalent cation, or a boron complex;
R8 is selected from H, aryl;
R9 is selected from substituted dihydropyridyl, alkyl of 1 to 10 carbon atoms, thioalkoxy of 1 to 10 carbon atoms, alkoxy of 1 to 10 carbon atoms, amino, cycloalkoxy of 3 to 4 carbon atoms, optionally substituted with one or more of amino, alkyl of 1 to 10 carbon atoms, alkylaryl of 1 to 10 carbon atoms, heteroaryl of 6 to 14 carbon atoms, alkylheteroaryl of 6 to 14 carbon atoms, alkylmercaptoalkyl of 1 to 10 carbon atoms, which may optionally be substituted with one or more of hydroxy, amino, guanidine, iminoalkyl of 1 to 10 carbon atoms;
X is selected from the group consisting of lower alkylenes of 2 to 10 carbon atoms, lower alkenylenes of 2 to 10 carbon atoms and lower alkynylenes of 2 to 10 carbon atoms and which may optionally be substituted by one or more alkyl of 1 to 10 carbon atoms, alkoxy of 1 to 4 carbon atoms, hydroxy, halogen, trifluoromethyl, nitro, cyano, or amino groups; or X is selected from the group consisting of the formula -(CH2)k Q(CH2)t- where k is 1, 2 or 3, t is 1, 2 or 3 and Q is O, Se, SiE2 where E is lower alkyl of 1 to 10 carbon atoms, aryl of 6 to 14 carbon atoms, S(O)g where g is 0, 1 or 2, or NR where R is H or lower alkyl of 1 to 10 carbon atoms which may be optionally substituted with lower alkyl of 1 to 10 carbon atoms, lower alkoxy of 1 to 4 carbon atoms, hydroxy, halogen, trifluoromethyl, nitro, cyano, amino; or X is selected from the group consisting of the formula -(CH2)m T(CH3)n- where m is 0, 1 or 2, n is 0, 1 or 2, T is a 3- to 6-membered carbocyclic or heterocyclic ring, aromatic ring or heteroaromatic ring which may optionally be substituted by one or more substituents selected from the group consisting of lower alkyl of 1 to 10 carbon atoms, lower alkoxy of 1 to 4 carbon atoms, hydroxy, halogen, nitro, cyano, trifluoroalkyl and amino.

2. A compound having the formula:
and pharmaceutically acceptable salts, wherein:
A is selected from O or S and may he taken together with R4 to form a heterocyclic ring of 3 to 6 carbon atoms; or A is N when R3 and R7 are taken together to form a heterocyclic ring of 3 to 6 carbon atoms; or R5 and A-R3 are taken together to form a covalent bond; or A is N-R3 provided R3 is not a heterocyclic radical of 3 to 6 carbon atoms;
R1 is selected from the group consisting of hydrogen, hydroxyalkyls of from 1 to 4 carbon atoms, alkoxyalkyls of from 1 to 4 carbon atoms in each position, lower alkyls of from 1 to 8 carbon atoms and haloalkyls of from 1 to 4 carbon atoms;
R2 is selected from the group consisting of straight and branched lower alkyls of from 1 to 4 carbon atoms, lower alkenyls of 2 to 10 carbon atoms and lower alkynyls of from 2 to 4 carbon atoms, cycloalkyls, cycloalkenyls of from 3 to 8 carbon atoms, and haloalkyls of from 1 to 4 carbon atoms;
is selected from the group consisting of aryl of 6 to 14 carbon atoms, heteroaryl, alkylaryl of 6 to 14 carbon atoms, alkylheteroaryl of 6 to 14 carbon atoms, all optionally substituted by one or more of halogen, nitrile, carboxy, carboxyalkyl of 1 to 10 carbon atoms, carboxyalkylaryl of 6 to 14 carbon atoms; or R3 is also selected from the group consisting of H, alkyl of from 1 to 4 carbon atoms, alkenyl of from 2 to 4 carbon atoms, CH2OC(=O)YR6, alkylhydroxy of 1 to 10 carbon atoms, alkylpolyhydroxy of 1 to 10 carbon atoms, alkyl(poly)oxyacyl of 1 to 10 carbon atoms, alkylcarboxy of 1 to 10 carbon atoms, optionally substituted by one or more of alkyl of from 1 to 4 carbon atoms, hydroxy, amino, carboxy, carboxyalkyl of 1 to 10 carbon atoms, alkylcarbonyl of 1 to carbon atoms;
R4 is selected from H. OH, SH, OR6, SR6, OC(=O)R6, SC(=O)R6, CH2OC(=O)YR6, OC(=O)YR6, SC(=O)YR6;
Y is independently selected from O, S, CH2, CHR6, C(R6)2, NH, NR6;
R5 is selected from H, OH, SH, OR6, SR6, OC(=O)R6, SC(=O)R6, CH2OC(=O)YR6, OC(=O)YR6, SC(=O)YR6;

provided that R4 and R5 are not simultaneously hydrogen or CH2OC(=O)YR6;
R6 is selected from hydrogen, alkyl of from 1 to 4 carbon atoms, alkenyl of 2 to 10 carbon atoms and alkynyl of from 2 to 4 carbon atoms, cycloalkyl of from 3 to 8 carbon atoms, heterocyclic radical of 3 to 6 carbon atoms, aryl, heteroaryl all optionally substituted by one or more alkyl of from 1 to 4 carbons atoms, alkoxy of 1 to 4 carbon atoms, hydroxy, halogen, trifluoromethyl, nitro, cyano, or amino groups;
R7 is selected from H, S(O)R9, SO2R9, CH2OC(O)-R9, C(O)-R9 where C(O)-R9 can represent natural and synthetic amino acids or R9 can be defined as below, or R7 and R3 taken together comprise a 5- or 6-membered heterocyclic ring containing two or more heteroatoms, optionally substituted with alkyl of from 1 to 4 carbon atoms and/or oxygen functions, or taken together comprise a metal complex containing a divalent cation, or a boron complex;
R8 is selected from H, acyl;
R9 is selected from substituted dihydropyridyl, alkyl of from 1 to 4 carbon atoms, thioalkoxy of 1 to 4 carbon atoms, alkoxy of 1 to 4 carbon atoms, amino, cycloalkoxy of 1 to 4 carbon atoms, optionally substituted with one or more of amino, alkyl of from 2 to 4 carbon atoms, alkylaryl of 5 to 14 carbon atoms, heteroaryl of 6 to 14 carbon atoms, alkylheteroaryl of 6 to 14 carbon atoms, alkylmercaptoalkyl of 1 to 10 carbon atoms, which may optionally be substituted with one or more of hydroxy, amino, guanidine, iminoalkyl of 1 to 10 carbon atoms;
x is selected from the group consisting of alkylenes, alkenylenes and alkynylenes having 2 to 6 carbon atoms and which may optionally be substituted by one or more alkyl groups having 1 to 10 carbon atoms; or X is selected from the group consisting of the formula -(CH2)k Q(CH2)t- where k is 1, 2 or 3, t is 1, 2 or 3 and Q is O, Se, SiE2 where E is lower alkyl of 1 to 10 carbon atoms, aryl of 6 to 14 carbon atoms, S(O)g where g is 0, 1 or 2, or NR where R is H or lower alkyl of 1 to 10 carbon atoms which may be optionally substituted with lower alkyl of 1 to 10 carbon atoms, lower alkoxy of 1 to 4 carbon atoms, hydroxy, halogen, trifluoromethyl, nitro, cyano, amino; or X is selected from the group consisting of the formula -(CH2)m T(CH2)n- where m is 0, 1 or 2, n is 0, 1 or 2, T is a 3- to 6-membered carbocyclic or heterocyclic ring, aromatic ring or heteroaromatic ring which may optionally be substituted by one or more substituents selected from the group consisting of lower alkyl of 1 to 10 carbon atoms, lower alkoxy of 1 to 4 carbon atoms, hydroxy, halogen, nitro, cyano, trifluoroalkyl and amino.

3. The compound as recited in Claim 2 wherein A is selected from O or S and may be taken together with R4 to form a heterocyclic ring; or A is N when R3 and R7 are taken together to form a heterocyclic ring; or R5 and A-R3 are taken together to form a covalent bond; or A is N-R3 provided R3 is not a heterocyclic radical;
R1 is selected from the group consisting of hydrogen, hydroxyalkyls of from 1 to 4 carbon atoms, alkoxyalkyls of from 1 to 4 carbon atoms in each position, lower alkyls of from 1 to 8 carbon atoms and haloalkyls of from 1 to 4 carbon atoms;
R2 is selected from the group consisting of straight and branched lower alkyls of from 1 to 4 carbon atoms, lower alkenyls and lower alkynyls of from 2 to 4 carbon atoms, cycloalkyls, cycloalkenyls of from 3 to 8 carbon atoms, and haloalkyls of from 1 to 4 carbon atoms;
R3 is selected from the group consisting of aryl, heteroaryl, alkylaryl, alkylheteroaryl, all optionally substituted by one or more of halogen, nitrile, carboxy, carboxyalkyl, and carboxyalkylaryl; or R3 is also selected from the group consisting of H, alkyl of from 1 to 4 carbon, atoms, alkenyl of from 2 to 4 carbon atoms, CH2OC(=O)YR6, alkylhydroxy, alkylpolyhydroxy, alkyl(poly)oxyacyl, alkylcarboxy;
R4 is selected from H, OH, SH, OR6, SR6, OC(=O)R4, SC(=O)R6, CH2OC(=O)YR6;
Y is independently selected from O, S, CH2, CHR6, C(R6)2, NH, NR6;
R5 is selected from H, OH, SH, OR6, SR6, OC(=O)R6, SC(=O)R6, CH2OC(=O)YR6;
provided that R4 and R5 are not simultaneously hydrogen;
R6 is selected from hydrogen, alkyl of from 1 to 4 carbon atoms, alkenyl and alkynyl of from 2 to 4 carbon atoms, cycloalkyl of from 3 to 8 carbon atoms, heterocyclic of from 5- to 8-members, aryl, heteroaryl all optionally substituted by one or more alkyl of from 1 to 4 carbon atoms or hydroxy groups;

R7 is selected from H, S(O)R9, SO2R9, CH2OC(O)-R9, C(O)-R9 where C(O)-R9 can represent natural and synthetic amino acids or R9 can be defined as below, or R7 and R3 taken together comprise a 5- or 6-membered heterocyclic ring containing two or more heteroatoms;
R8 is selected from H, acyl;
R9 is selected from substituted dihydropyridyl, alkyl of from 1 to 4 carbon atoms, thioalkoxy, alkoxy, amino, cycloalkoxy;
X is selected from the group consisting of alkylenes, alkenylenes and alkynylenes having 2 to 6 carbon atoms and which may optionally be substituted by one or more alkyl groups; or X is selected from the group consisting of the formula -(CH2)k Q(CH2)t- where k is 2 or 3, t is 1 or 2 and Q is O, Se, SiE2 where E is lower alkyl, aryl, S(O)g where g is 0, 1 or 2, or NR where R is H or lower alkyl which may be optionally substituted with lower alkyl as defined in claim 1; or X is selected from the group consisting of the formula -(CH2)m T(CH2)n- where m is 0, 1 or 2, n is 0, 1 or 2, T is a 3- to 6-membered carbocyclic or heterocyclic ring, aromatic ring or heteroaromatic ring which may optionally be substituted by one or more substituents selected from the group consisting of lower alkyl, lower alkoxy, hydroxy, halogen, nitro, cyano, trifluoroalkyl and amino.

4. The compound as recited in Claim 3 wherein A is selected from O and may be taken together with R4 to form a heterocyclic ring; or A is N when R5 and A-R3 are taken together to form a covalent bond;~

A is N-R3 where the R3 radicals are selected from hydrogen, alkyl of from 1. to 4 carbon atoms or aryl;

R1 is selected from the group consisting of hydrogen, hydroxyalkyls of from 1 to 4 carbon atoms and lower alkyls of from 1 to 8 carbon atoms;

R2 is selected from the group consisting of straight and branched lower alkyls of from 1 to 4 carbon atoms and haloalkyls of from 1 to 4 carbon atoms;

R3 is selected from the group consisting of aryl, heteroaryl, alkylaryl, alkylheteroaryl, all optionally substituted by one or more of halogen, nitrile, carboxy, carboxyalkyl, carboxyalkylaryl; or R3 is also selected from the group consisting of H, alkyl of from 1 to 4 carbon atoms and alkenyl of from 2 to 4 carbon atoms;

R4 is selected from H, OH, SH, OR4, SR6;

R5 is selected from H, OH, SH, OR6, SR6;

provided that R4 and R5 are not simultaneously hydrogen;

R6 as selected from hydrogen, alkyl of from 1 to 4 carbon atoms, alkenyl and alkynyl o~ from 2 to 4 carbon atoms and cycloalkyl of from 3 to a carbon atoms;

R7 is selected from H and where C(O)-R9 can represent natural and synthetic amino acids;

R8 is selected from H and acyl;

X is selected from the group consisting of alkylenes, alkenylenes and alkynylenes having 2 to 6 carbon atoms and which may optionally be substituted by one or more alkyl groups; or X is selected from the group consisting of the formula -(CH2)k Q(CH2)t- where k is 2 or 3, t is 1 or 2 and Q is O, S(O)g where g is 0, 1 or 2, or NR where R is H or lower alkyl; or X is selected from the group consisting of the formula -(CH2)m T(CH2)n- where m is 0, 1. or 2, n is 0, 1 or 2, T
is a 3- to 6-membered carbocyclic or heterocyclic ring, aromatic ring or heteroaromatic ring.

5. The compound as recited in Claim 4 wherein A is O;
R1 is hydrogen;
R2 is methyl;
R3 is selected from the group consisting of hydrogen, and lower alkyls of 1 to about 4 carbon atoms;
R4 is hydroxy;
R5 is hydrogen or hydroxy;
R7 is hydrogen;
R8 is hydrogen;
X is an alkylene having 3 to 5 carbon atoms.

6. ~The pharmaceutical composition as recited in claim 3 wherein said compound is selected from the group consisting of:

N6-[1-(hydroxyimino)ethyl]-L-lysine, dihydrochloride 2-amino-5-[[1-{hydroxyimino)ethyl]amino]-2-methylpentanoic acid, dihydrochloride N6-[1-(hydroxyimino)ethyl]-2-methyl-L-lysine, dihydrochloride 2-[[2-[[1-{hydxoxyimino)ethyl]amino]ethylseleno]methyl]-L-alanine, dihydrochloride N6-[1-{hydroxyimino)ethyl]-2-(hydroxymethyl)-L-lysine, dihydrochloride N6-[1-(hydroxyimino)-2,2,2-trifluoroethyl]-L-lysine, dihydrochloride N6-[1-(hydroxyimino)ethyl]-L-lysine, ethyl ester, dihydrochloride N6-[1-(hydroxyimino)ethyl]-L-lysine, 2,3-dihydroxypropyl ester, dihydrochloride N6-[1-(hydroxyimino)ethyl]-L-lysine, 2-hydroxy-1-(hydroxymethyl)ethyl ester, dihydrochloride N6-[1-(hydroxyimino)ethyl]-L-lysine, 2-(diethylamino)-2-oxoethyl ester, dihydrochloride S-ethyl 2S-amino-6-[[1{hydroxyimino)ethyl]amino]hexanethioate, dihydrochloride N2-[(acetyloxy)methyl]-N6-[1-(hydroxyimino)ethyl]-L-lysine, dihydrochloride N6-[1-(hydroxyimino)ethyl]-N2-[[[(methylamino)carbonyl]oxy]methyl]-L-lysine, dihydrochloride N6-[1-(hydroxyimino)ethyl]-N2-[[(methoxycarbonyl)oxy]methyl]-L-lysine, dihydrochloride N6-(1-(hydroxyimino)ethyl]-N2-[[[(methylthio)carbonyl]oxy]methyl]-L-lysine, dihydrochloride N6-[1-(hydroxyimino)ethyl]-N2-([(phenylcarbonyl)oxy]methyl]-L-lysine, dihydrochloride N2-acetyl-N2-[(acetyloxy)methyl]-N6-[1-(hydroxyimino)ethyl]-L-lysine, hydrochloride N6-[1-(hydroxyimino)ethyl]-N2-[(methylthio)carbonyl]-L-lysine, hydrochloride N6-[(1,1-dimethylethoxy)carbonyl]-N6-[1-(hydroxyimino)ethyl]-L-lysine, hydrochloride N6-[1-(hydroxyimino)ethyl]-N2-[(methylamino)carbonyl]-L-lysine, hydrochloride N6-[1-(hydroxyimino)ethyl]-N2-[N6-[1-(hydroxyimino)ethyl]-L-lysyl]-L-lysine, ethyl ester, trihydrochloride N6-[2-(hydroxyimino)ethyl]-N2-[N6-[2-(hydroxyimino)ethyl]-L-lysyl]-L-lysine, trihydrochloride N6-[1-(hydroxyimino)ethyl]-N2-[N6-(1-iminoethyl)-L-lysyl]-L-lysine, ethyl ester, trihydrochloride N6-[1-(hydroxyimino) ethyl]-N2-[N6-(1-iminoethyl)-L-lysyl]-L-lysine, trihydrochloride N2-(N-acetyl-L-methionyl)-N6-[1-(hydroxyimino)ethyl]-L-lysine, ethyl ester, hydrochloride N2-(N-acetylmethionyl)-N6-[1-(hydroxyimino)ethyl)-L-lysine, hydrochloride N2-(L-alanyl}-N6-[1-(hydroxyimino)ethyl]-L-lysine, dihydrochloride N6-[1-[((methoxycarbonyl)oxy]imino]ethyl]-L-lysine, dihydrochloride N6-[1-[([(methylamino)carbonyl]oxy]imino]ethyl]-L-lysine, dihydrochloride N6-[1-[[[(methylthio)carbonyl]oxy]amino]ethyl]-L-lysine, dihydrochloride N6-[(1-[(acetyloxy)imino]ethyl]-L-lysine, dihydrochloride N6-[1-[[(phenylcarbonyl)oxy]imino]ethyl]-L-lysine, dihydrochloride N6-[1-[(methoxyimino)ethyl]-L-lysine, dihydrochloride N6-[1-[(phenoxyimino)ethyl]-L-lysine, dihydrochloride N6-hydroxy-N6-[1-(hydroxyimino)ethyl]-L-lysine, dihydrochloride N6-(1-(hydroxyimino)ethyl]-N6-([(methoxycarbonyl)oxy]methyl]-L-lysine, dihydrochloride N6-(1-(hydroxyimino)ethyl]-N6-([[(methylamino)carbonyl]oxy]methyl]-L-lysine, dihydrochloride N6-[1-(hydroxyimino)ethyl]-N6-[[[(methylthio)carbonyl]oxy]methyl]-L-lysine dihydrochloride N6-[1-(hydroxyimino)ethyl]-N6-[[(phenylcarbonyl)oxy]methyl]-L-lysine, dihydrochloride N6-(acetyloxy)-N6-(1-iminoethyl)-L-lysine, dihydrochloride N6-[1-((phenylthio)imino]ethyl]-L-lysine, dihydrochloride N6-(1-iminoethyl)-N6-(phenylmethoxy)-L-lysine, dihydrochloride N6-hydroxy-N6-(1-iminoethyl)-L-lysine, dihydrochloride N6-acetyloxy-N6-[1-[(acetyloxy)imino]ethyl]-L-lysine,~
dihydrochloride N6[1-[[1(methoxycarbonyl)oxy]imino]ethyl]-N6-[(methoxycarbonyl)oxy]-L-lysine, dihydrochloride N6-[1-[[[(methylamino)carbonyl]oxy]imino)ethyl]-N6-[[(methylamino)carbonyl]oxy)-L-lysine, dihydrochloride N6-[1-[[[(methylthio)carbonyl]oxy)imino]ethyl]-N6-[[(methylthio)carbonyl]oxy]-L-lysine, dihydrochloride N6-[1-[[(phenylcarbonyl)oxy]imino]ethyl]-N6-[(phenylcarbonyl)oxy]-L-lysine, dihydrochloride N6-[1-(phenoxyimino)ethyl]-N6-phenoxy-L-lysine, dihydrochloride N6-[1-(methoxyimino)ethyl]-N6-(phenylmethoxy)-L-lysine, dihydrochloride N6-[1-[[(acetyloxy)methyl]imino]ethyl]-N6-hydroxy-L-lysine, dihydrochloride N6-hydroxy-N6-[1-[[[(methoxycarbonyl)oxy]methyl]imino]ethyl]-L-lysine, dihydrochloride N6-hydroxy-N6-[1-[[[[(methylamino)carbonyl]oxy]methyl]imino]ethyl]-L-lysine, dihydrochloride N6-hydroxy-N6-[1-[[[((methylthio)carbonyl]oxy]methyl]imino]ethyl]-L-lysine, dihydrochloride N6-hydroxy-N6-[1-[[[(phenylcarbonyl)oxy]methyl)imino]ethyl]-L-lysine, dihydrochloride N6-[(acetyloxy)methyl]-N6-[1-(hydroxyimino)ethyl]-L-lysine, dihydrochloride N6-(1-iminoethyl)-N6-((methoxycarbonyl)oxy]-L-lysine, dihydrochloride N6-(1-iminoethyl)-N6-[[(methylamino)carbonyl]oxy]-L-lysine, dihydrochloride N6-(1-iminoethyl)-N6-[[(methylthio)carbonyl]oxy]-L-lysine, dihydrochloride N6-(1-iminoethyl)-N6-[(phenylcarbonyl)oxyl-L-lysine, dihydrochloride N6-(1-iminoethyl)-N6-phenoxy-L-lysine, dihydrochloride N6-[1-[(methylthio)imino]ethyl]-L-lysine, dihydrochloride N6-(1-iminoethyl)-N6-(methylthio)-L-lysine, dihydrochloride N6-[1-iminoethyl)-N6-(phenylthio) -L-lysine, dihydrochloride N6-(1-[(phenylthio)imino]ethyl]-L-lysine, dihydrochloride 5-[4-[[1-(hydroxyimino)ethyl)amino]butyl]imidazolidine-2,4-dione, dihydrochloride N-[N6-[1-(hydroxyimino)ethyl]-L-lysyl]-L-alanine, dihydrochloride 2S-amino-6-[[1-(hydroxyimino)ethyl]amino]-4-hexenoic acid, dihydrochloride N-[2S-amino-2-[3-[[[1-(hydroxyimino)ethyl]amino]methyl]cyclopentyl]acetyl]-L-alanine, dihydrochloride N-[2S-amino-2-[3-[[[1-(hydroxyimino)ethyl]amino]methyl]isoxazol-5-yl]acetyl]-L-alanine, dihydrochloride N2-(4-amino-1-oxobutyl)-N6-[1-(hydroxyimino)ethyl]-L-lysine, ethyl ester, dihydrochloride N2-[[(2-aminoethyl)amino]carbonyl]-N6-[1-(hydroxyimino)ethyl]-L-lysine, ethyl ester, dihydrochloride N2-[(2-aminoethoxy)carbonyl]-N6-[1-(hydroxyimino)ethyl]-L-lysine, ethyl ester, dihydrochloride 9S-amino-5,6,7.8,9,10-hexahydro-3-methyl-4H-1,2,4-oxadiazecin-10-one, dihydrochloride 3S-aminohexahydro-1-[1-(hydroxyimino)ethyl]-2H-azepin-2-one, dihydrochloride

7. Use of a therapeutically effective amount of a compound according to any one of claims 1, 2, 3, 4, 5 or 6 for inhibiting nitric oxide synthesis in a subject in need of such inhibition.

8. Use of a therapeutically effective amount of a compound of claims 1, 2, 3, 4, 5, or 6 for selectively inhibiting citric oxide synthesis produced by inducible NO
synthase over nitric oxide produced by the constitutive forms of NO synthase in a subject in need of such selective inhibition.

9. Use of a therapeutically effective amount of a compound of claims 1, 2, 3, 4, 5, or 6 for lowering nitric oxide levels in a subject in need of such treatment.

10. A pharmaceutical composition comprising a compound of claims 1, 2, 3, 4, 5, or 6 together with one or more pharmaceutically acceptable carriers.