CA2184691C - L-n6-(1-iminoethyl)lysine derivatives useful as nitric oxide synthase inhibitors - Google Patents

Abstract

There is disclosed a novel amino glycol derivatives of L-N6-(1-iminoethyl)lysine, pharmaceutical compositions containing these novel compounds, and to their use in therapy, in particular their use as nitric oxide synthase inhibitors.

Description

ZiiN~-ll-IMINOETHYL LYSINE DERIVATIVES USEFUL AS
NITRIC OXIDE SYNTHASE INHIBITORS
~ck~ronnd of the Invention 1~ F,'_e1_d of the Invention The present invention relates to novel amino glycol-derivatives of Z-N6-(1-iminoethyl)lysine, pharmaceutical compositions. containing these novel compounds, and to their 15 use in therapy, in particular their use as nitric oxide synthase inhibitors.

2 ~ It has been known since the early 1980's that the vascular relaxation brought about by acetycholine is dependent on the presence of the endothelium and this activity was ascribed to a labile humoral factor termed endothelium-derived relaxing factor (EDRF). The activity of 2 5 nitric oxide (NO) as a vasodilator has been known for well over 100 years and NO is the active component of amylnitrite, glyceryltrinitrite and other nitrovasodilators.
The recent identification of EDRF as NO has coincided with the discovery of a biochemical pathway by which NO is 3 0 synthesized from the amino acid Z-arginine by the enzyme NO synthase.
NO is the endogenous stimulator of the soluble guanylate cyclase and is involved in a number of biological 3 5 actions in addition to endothelium-dependent relaxation including cytotoxicity of phagocytic cells and cell-to-cell communication in the central nervous system (see Moncada et WO .95/24382 PCT/US95/02669 21~4b91 a1 j Biochemical Pharmacology,- 38, 1709-1715 (1989) and Monc-a_da et ale, Pharmacological Reviews. 43, 109-142 (1991).
It is now thought that excess NO 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 analogue, L-N-monomethyl-arginine (L-NMMA) and the therapeutic use of L-NN~IA 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-0446599.
It has recently become apparent that there are at least three types of NO synthase as follows:
(i) a constitutive, Ca++/calmodulin dependent enzyme, 2 0 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.
2 5 (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 3 5 microorganisms. It also appears that the adverse effects of WO 95/24382 218 4 b 91 PCT/US95/02669 excess NO production, in particular pathological vasodilation and tissue damage, may result largely from the effects of NO synthesized by the inducible NO synthase.
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 autoimmune and/or inflammatory conditions affecting the joints, for example arthritis, inflammatory bowel disease, cardiovascular ischemia, diabetes, hyperalgesia (allodynia), cerebral ischemia (both focal ischemia, thrombotic stroke and global ischemia, secondary to cardiac arrest), and other CNS disorders mediated by NO.
Futher conditions in which there is an advantage in inhibiting NO production from L-arginine include systemic 2 0 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.
2 5 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 3 0 order to avoid the potentially serious consequences of over-inhibition of the constitutive NO-synthase including hypertension and possible thrombosis and tissue damage. In particular, in the case of the therapeutic use of L-NMMA for the treatment of toxic shock it has been recommended that 3 5 the patient must be subject to continuous blood pressure a CA 02184691 2004-05-26 monitoring throughout the treatment. Thus, while non-selective NO synthase inhibitors have therapeutic utility provided that appropriate precautions are taken, NO synthase inhibitors which are selective in the sense that they S inhibit the inducible NO synthase to a considerably greater extent than the constitutive isoforms of NO synthase would be of even greater therapeutic benefit and easier to use.
W094/12165, W094/14780, W093/13055, EP0446699A1 and U.S. Patent No. 5,132,453 disclose compounds that inhibit nitric oxide synthesis and preferentially inhibit the inducible isoform of nitric oxide synthase.
In accordance with the present invention novel amino glycol derivatives of L-N6-(1-iminoethyl)lysine derivatives 2 0 are provided. These novel inhibitor compounds can be represented by the following chemical formula. A compound or a pharmaceutically acceptable salt, prodrug or ester therof having the formula:
Ra N B
A
Y 'N
~3 R

Y is a hydrogen, lower alkyl radical, lower alkeryl radical, lower alkynyl radical, aromatic hydrocarbon radical, alicyclic hydrocarbon.radical, amino, heterocyclyl radical WO 95/24382 218 4 6 91 PCT~S95/02669 in which 1 to about 4 heteroatoms are independently selected from oxygen, nitrogen and sulfur, wherein all said radicals may optionally be substituted with hydrogen, cyano, lower alkyl, nitro, amino, alicyclic hydrocarbon radicals, or 5 aromatic hydrocarbon radicals which may be optionally substituted with lower alkyl;
X is lower alkyl radical, lower alkenyl radical, lower alkynyl radical, aromatic hydrocarbon radical, 1 0 (CH2) mQ (CH2) n, where m= 1-3, n = 1-3, and Q is sulfur, sulfinyl, sulfonyl or oxygen, C=0, lower alkynyl radical, aromatic hydrocarbon radical, alicyclic hydrocarbon radical or heterocyclyl radicals in which 1 to about 4 heteroatoms are independently selected from oxygen, nitrogen and sulfur, wherein all said radicals are optionally substituted with hydrogen, halogen and lower alkyl;
Rl, R2, R3 and R4 are independently selected from the group consisting of hydrogen and lower alkyl;
A is a lower alkyl radical, lower alkenyl radical, lower alkynyl radical, alicyclic hydrocarbon radical, C=O, aromatic hydrocarbon radical or heterocyclyl radical in which 1 to about 4 heteroatoms are independently 2 5 selected from oxygen, nitrogen and sulfur, wherein all said radicals are optionally substituted with hydrogen, lower alkyl, hydroxyl, lower alkoxy, alkoxycarbonyl, alkylaryloxy, thiol, lower thioalkoxy, thioalkylaryloxy, thioaryloxy, sulfinylalkyl, sulfinylalkylaryl, sulfinylaryl, sulfonylalkyl, sulfonylalkylaryl, sulfonylaryl, halogen, aromatic hydrocarbon radicals, or alicyclic hydrocarbon radicals;
B can be hydrogen, lower alkyl radical, lower alkenyl radical, lower alkynyl radical, lower alkoxy radical, WO 95124382 218 4 6 91 pCT/US95/02669 hydroxy, alkoxycarbonyl, alkylaryloxy, thiol, lower thioalkoxy, lower thioalkylaryloxy, thioaryloxy, sulfinylalkyl, sulfinylalkylaryl, sulfinylaryl, sulfonylalkyl, sulfonylalkylaryl, sulfonylaryl, aromatic hydrocarbon radical, alicyclic hydrocarbon radical, or heterocyclyl radical in which 1 to about 4 heteroatoms are independently selected from oxygen, nitrogen and sulfur wherein all said radicals are optionally substituted with hydrogen, lower alkyl, hydroxyl, lower alkoxy, halogen, aromatic hydrocarbon radicals, or alicyclic hydrocarbon radical, or B can be C (=0) ORS, C (=O) NR5R6, P (=0) (0R5) (0R6) , NHOH, N (OH) C (=O) NR5R6, NRSC (=O) NR6R~, NRSC (=O) N (OH) R6, C (=0) NHOH, where R5 is hydrogen, lower alkyl radical, aromatic hydrocarbon radical, or alicyclic hydrocarbon radical wherein all said radicals are optional substituted with lower alkyl, lower 2 0 alkenyl;
R6 is hydrogen, lower alkyl radical, aromatic hydrocarbon radical, or alicyclic hydrocarbon radical wherein all said radicals are optional substituted with lower alkyl, lower 2 5 alkenyl; and R~ is hydrogen, lower alkyl radical, aromatic hydrocarbon radical, or alicyclic hydrocarbon radical wherein all said radicals are optional substituted with lower alkyl, lower 3 0 alkenyl;
with the proviso that when A is C=0, B may not be hydroxy or alkoxy.
(I) WO 95/24382 218 4 6 91 pCT~S95/02669 A is preferably lower alkyl which is substituted as indicated above.
In another broad aspect, the present invention is directed to inhibiting nitric oxide synthesis in a subject in need of such inhibition or treatment by 2.dministering a compound of Formula (I) which preferentially inhibits the inducible isoform of nitric oxide synthase over the constitutive isoform of nitric oxide synthase, in a nitric oxide synthesis inhibiting amount to such subject.
The invention further relates to a pharmaceutical composition comprising a compound from Formula (I).
Compounds and compositions defined above have usefulness as inhibitors of nitric oxide synthase. These compounds also preferentially inhibit the inducible form over the constitutive form by at least 3 fold.
Conditions in which there is an advantage in inhibiting 2 0 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. Further conditions in which there is 2 5 an advantage in inhibiting NO production from L-arginine include autoimmune diseases and/or inflammatory conditions such as those affecting the joints, for example arthritis or inflammatory bowel disease, cardiovascular ischemia, diabetes, cerebral ischemia and other CNS disorders mediated 3 0 by NO .
A preferred embodiment of the present invention is a compound of the formula (I) wherein Y is hydrogen or lower alkylene 3 S X is lower alkylene from 3-5 carbon R1, R2, R3, and R4 are independently selected from the group consisting of hydrogen or lower alkyl A is lower alkylene from 2-4 carbons sustituted with hydroxyl B is hydroxyl.
It is preferred that Y is methyl, X is preferably butylene, R1, R2, R3, and R4 are preferably hydrogen, A is preferably ethylene or isopropylene substituted with hydroxyl and B is preferably hydroxyl (OH).
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.
2 0 Thus, preferred salts include those formed from hydrochloric, hydrobromic, sulfuric, citric, tartaric, phosphoric, lactic, acetic, succinic, fumaric, malefic, methanesulfonic, ethanesulfonic,p-toluenesulfonic, benzenesulfonic and the like. (See, for example, S. M.
2 5 Berge et al., Pharmaceutical Salts, J. Pharm. Sci., 1977, 66, 1-19.) 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.
3 0 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 formulation. According to a further aspect, the present invention provides a pharmaceutical formulation comprising a compound of formula 35 (I) or a pharmaceutically acceptable salt or solvate WO 95/24382 218 4 6 91 pCT/US95/02669 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.
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 2 0 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 finely divided solid carriers or both and then, if necessary, shaping the product into the desired 2 S 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 3 0 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.

WO 95/24382 218 4 6 91 P~'~595/02669 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 5 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 by moulding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent. The 10 tablets may optionally be coated or scored and may be formulated so as to provide slow or controlled release of the active ingredient therein.
Formulations for parenteral administration include aqueous and non-aqueous sterile injection solutions which may contain antioxidants, buffers, bacteriostats and 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 2 0 thickening agents. The formulations may be presented in unit-dose or multi-dose containers, for 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-2 S injection, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets of the kind previously described.
3~ 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, 3 5 for example buccally or sublingually, include lozenges WO 95/24382 218 4 6 91 pCT~S95/02669 comprising the active ingredient in a flavored 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 hereinbelow recited, or an appropriate fraction thereof, of the active ingredient.
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 flavoring 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 2 0 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 dosage or as a multiple of the same, for instance, units containing 5 mg to 500 mg, usually around 10 2 5 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 3 0 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 o~ administration may vary depending on the 3 5 condition and its severity.

WO 95/24382 PCT/US95l02669 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.
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 2 0 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 2 5 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 "alicyclic hydrocarbon" or "cycloalkyl" means a aliphatic radical in a ring with 3 to about 10 carbon 3 0 atoms, and preferably from 3 to about 6 carbon atoms.
Examples of suitable alicyclic radicals include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl and the like.
The term "aromatic hydrocarbon radical" means 4 to 3 5 about 16 carbon atoms, preferably 6 to about 12 carbon atoms, more preferably 6 to about 10 carbon atoms. Examples of suitable aromatic hydrocarbon radicals include phenyl,naphthyl, and the like.
The term "aryl" as used herein means 5- and 6-membered single-aromatic radicals which may include from zero to four heteroatoms. Representative aryls include phenyl, thienyl, furanyl, pyridinyl, (is)oxazoyl and the like.
The term DCM means dichioromethane.
The term DEAD means diethyl azodicarboxylate.
The term DIBAL-H means diisobutylaluminum hydride.
The term DMAP means dimethylaminopyridine.
The term DMSO means dimethylsulfoxide.
The term EDC means 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride.
The term "heterocyclyl radical" means a saturated or unsaturated cyclic hydrocarbon radical including aromatic systems with 4 to about 10 carbon atoms, preferably about 5 to about 6; wherein 1 to about 4 carbon atoms are replaced by nitrogen, oxygen or sulfur. The "heterocyclic radical"
2 0 may be fused to an aromatic hydrocarbon radical. Suitable examples. include pyrrolyl, pyridinyl, pyrazolyl, triazolyl, pyrimidinyl, pyridazinyl, oxazolyl, isoxazolyl, thiazolyl, imidazolyl, indolyl, thienyl, furanyl, tetrazolyl, 2-pyrrolinyl, 3-pyrrolinyl, pyrrolindinyl, 1,3-dioxolanyl, 2-2 5 imidazonlinyl, imidazolidinyl, 2-pyrazolinyl, pyrazolidinyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiadiazolyl, 2H-pyranyl, 4H-pyranyl, piperidinyl, 1,4-dioxanyl, morpholinyl, 1,4-dithianyl, thiomorpholinyl, pyrazinyl, piperazinyl, triazinyl, 1,3,5-trithianyl, 30 benzo(b)thiophenyl, benzimidazolyl, quinolinyl, and the like.
The term HOBT means N-hydroxybenzotriazole.
The term "lower alkoxy", alone or in combination, means an alkyl ether radical wherein the term alkyl is as defined 3 S above and most preferably containing 1 to about 4 carbon o ~ CA 02184691 2004-05-26 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 term "lower thioalkoxy", alone or in combination, means an alkyl thioether radical wherein the term alkyl is as defined above and most preferably containing 1 to about 4 carbon atoms. Examples of suitable alkyl thioether radicals include thiomethoxy, thioethoxy, thio-n-propoxy, thio-i-propoxy, thio-n-butoxy, thio-iso-butoxy, thio-sec-butoxy;
thio-tert-butoxy and the like.
The term alkoxycarbonyl as used herein means an alkoxy group, as defined above, having a carbonyl (C=O) group attached.
The term "halogen" means fluorine, chlorine, bromine or 1 5 iodine .
The term mcpba means m-chloroperbenzoic acid.
The term NMM means N-methylmorpholine.
The term NMMO means 4-methylmorpholine N-oxide.
The term "prodrug" refers to a compound that is made 2 0 more active in vivo.
The term sulfinyl means S0.
The term sulfonyl means 502, The term TEA means triethylamine.
The term TMSN3means azidotrimethylsilane.
2 5 As used herein, reference to "treatment" of a patient ' is intended to include prophylaxis.
3 0 Compounds of the present invention can exist in geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis- and trans-geometric isomers, E- and Z-geometric isomers, R- and S-enantiomers, diastereomers, d-isomers, 1-isomers, the WO 95/24382 218 4 6 91 pCT~S95/02669 racemic mixtures thereof and other mixtures thereof, as falling within the scope of the invention.
Disclosed are twenty eight general synthetic processes 5 useful in the preparation of the compounds of the present invention.
Scheme 1 BocHN-(CH2)m-X-(CH2) COOH a ~ gocHN-(CH2)m-X-(CH2) COCHN2 ZNH ZNH

b BocHN-(CH2)m-X-(CH2) CH COOR

(a) (i) i-butyl chloroformate, NMM, THF; (ii) CH2N2; (b) Ag benzoate, R50H, TEA.

Scheme 2 BocHN-(CH2)m-X-(CH2) COOH a CON(OMe)Me _~ BocHN-(CH2)m-X-(CH2)n~
ZNR~ ~ Z~(N'R~
BocHN-(CH2)m-X-(CH2) COB
ZNR
(a) N,O-Dimethylhydroxylamine HCI, EDC, HOBT, TEA, DMF; (b) B'Li, THF.

Scheme 3 ~\ O OH
H -.~. I / b,~.

\ O O R d ~\ O ORs NHBoc NHBoc (a) ammonium acetate, malonic acid, acetic acid; (b) di-t-butyl Bicarbonate, NaOH/dioxane;
(c) NaHC03, DMF, R51; (d) H2 PdIC.
Scheme 4 \ -.~ ~\ O OH b c, ~\ O OR5 I / H I / a ' I /
T
O NH2 NHBoc (a) ammonium acetate, malonic acid, acetic acid; (b) di-t-butyl Bicarbonate, NaOHldioxane;
(c) NaHC03, DMF, R$1; (d) H2 Pd/C.
Scheme 5 ....~1 H H
N~ a, b~ ,N A c~ d° H.N A
H.-.~O Boc ~ e~' O NHCbz (a) di-t-butyl Bicarbonate, DMAP, THF; (b) A*Li; (c) hydroxylamine hydrochloride (d) H2 PdIC ~e) CbzCl (f) HClldioxane.

Scheme 6 A H H
N JP a,~ N NH - d=~ Boc'N NH2 H O Boc' A O A
(a) di-t-butyl Bicarbonate, DMAP, THF; (b) LiOH; (c) i-butyl chloroformate, ammonia (d) trifluoroacetic anhydride, Et3N (e) H2/Pd.
Scheme 7 Br O(OR5)2 BocHN-(CH2)m-O-(CH2) a BocHN-(CH2)m-O-(CH2) NR~Z NR~Z
(a) P(OR$)3.

WO 95124382 218 4 6 91 pCT~S95/02669 Scheme 8 OMe OH
BocHN-(CH2)m-O-(CH2) O a BocHN-(CH2)m-O-(CH2) NR~Z NR~Z
Br _ b BocHN-(CH2)m-O-(CH2) c NR~Z
NPhth d BocHN-(CH2)m-Q-(CH2) BocHN-(CH2)m-Q-(CH2) NR~Z
NR~Z
(a) DIBAL-H, toluene, -70°C, 1 h; (b) Ph3PBr2, DMAc, 16 h; (c) K+NPhth, THF;
(d) NH2NH2, EtOH.
Scheme 9 NHRs ~O

BocHN-(CH2)m-Q-(CH2) a BocHN-(CH2)m-Q-(CH2) NR~Z NR~Z
(a) O=C=NR6, DCM.

Scheme 10 NOBzIRs ~0 BocHN-(CH2)m-Q-(CH2) a BocHN-(CH2)m-O-(CH2) NR~Z NR~Z
(a) carbonyldiimidiazole, NHOBzIRs, DCM.
Scheme 11 B B

A CH A
~(CHZ)~ a BocHN-(CH2)rrr~( NR~Z NR~Z
(a) n-BuLi, THF, BocNH(CH2)mBr.
Scheme 12 HS-(CH2) A a I BocHN- CH m-S- CH A
( z) ( z) NR~Z NR~Z
(a) NaOMe, THF, BocNH(CH2)mBr.

WO 95/24382 21$ 4 b 9 l PCT/US95/02669 Scheme 13 SerOMe BocHN-(CH2)m-O-(CH2) O a NHZ
O ~ O
BocHN-(CH2)m-O-(CH2) \N
NHZ OEt Ph3P, CBr4, (ii) Cu Scheme 14 B B
ZHN-(CH2)m-O-(CH2) A ~HOAc~H2N-(CH2)m-O-(CH2) A
BocNR BocNR~
NuH B
H2N N (CH2)m-O-(CH2)r~A ~-..~ H2N~N-(CH2)m-O-(CH2)n~A
HCI NR~ BoC HCI NR~ H
HCI
(a) H2, Pd black, EtOH/HOAc; (b) (i) 3,5-dimethylpyrazole-1-carboxamidine nitrate, NaOH;
(ii) 1 N HCI; (c) HCUdioxane, HOAc.

WO 95/24382 ~ ~ 8 4 6 91 PCT/US95/02669 Scheme 15 B
BocHN-(CH2)m-Q-(CH2) A a A
---~ HCI~H2N-(CH2)m-Q-(CH2) ZNR
b NH g NH g NH(CH2)m-Q-(CH2) A c ~ NH(CH2)m-O-(CH2) A
HCI ' HCI
ZNR~ HNR1 HCI
(a) HCUdioxane, HOAc; (b) (i) methyl acetimidate, NaOH; (ii) 1 N HCI;
(c) H2, Pd black, EtOH, HOAc.
Scheme 16 NH
NH(CH2)m-Q-(CHI CH2COOMe a HCI
HNR~
NH
NH(CH2)m-O-(CH~j CH2COOH
HCI
HNR~
HCI
(a) 2 N HCI, ~.

WO 95/24382 218 4 6 91 PCT~S95/02669 Scheme 17 BocHN BocHN
a Z~N / ~ Z. / S
H j-I I /
BocHN OH BocHN OH
~Z.N S _~ H.
i H OH ~ H OH I /
NH BocHN OH NH NH2 OH
~ ~N S --i~N S

H OH ~ H 2 HCl OH I /
(a) Pd(OAc)2ltri-o-tolylphosphine/2-bromothiopheneRriethylamine; (b) Os04, NMMO, acetone-H20; (c) H2/Pd/AcOH; (d) ethyl acetimidate HCI/EtOH; (e) HCI/Dioxane/AcOH; (f) HCIM20.

Scheme 18 BocHN BocHN
/ a H
---~ Z.N / N
H H I
N
BocHN OH N c H' BocHN OH N
Z~ H --_ _ OH ~ ~ H _ -- OH I
N
NH BocHN OH H e, f NH NH2 OH H
N .--t ~ N
- _ OH ~ ~ H _ -- OH I
3 HCl N
(a) Pd(OAc)2/tri-o-tolylphosphine/4-bromoimidazole/triethylamine; (b) Os04, NMMO;
(c) Hz/Pd/AcOH; (d) ethyl acetimidate HCVEtOH; (e) HCVDioxane/AcOH; (f) HCVH20.

Scheme 19 a N OH ---~ ~ N O
H OH
H 2 HC1 O~ Me b Me N O
I
H O H
2 TsOH
(a) 2,2-dimethoxypropane/DMF/HCI/Dioxane; (b) 1,1,1-trichloroethane/TsOH/benzaldehyde.

Scheme 20 NHBOC NHBOC Me ZwN a Z.N O OMe I I
H ~ H pH O
NHBOC Me HwN O~OMe c~
I
H OH O
NH NHBOC Me ~N O OMe I
H OH O
NH NH2 Me ~N O OMe ---I
H 2 HCl OH O
NHp N O
I
H 2 HCl O Me O
(a) NaH/DMF2nC12/THF/methyl lactate; (b) H2/Pd/AcOH; (c) ethyl acetimidate HCUEtOH;
(d) HCUDioxane/AcOH; (e) HCUH20.

WO 95/24382 218 4 6 91 pCT/US95/02669 Scheme 21 NHBOC OTHP NHBOC pTHP
Z \ Br a Z
+ --'N O I / 'N I \
H H OH /
NHBOC OTHP NH NHBOC OTHP
H ~ N \ ---' ~N \
I
H OH ~ H OH I

~N \

(a) n-BuLi/THF/-78 °C; (b) H~/Pd/AcOH; (c) ethyl acetimidate HCI/EtOH;
(d) HCNDioxane/AcOH.

WO 95/24382 218 4 6 91 P~~1S95/02669 Scheme 22 BocHN-(CH2)m-X-(CH2)~ ON(OMe)Me ~ BocHN-(CH2)m-X-(CHZ) CHO
~ZNH ZNH
R
b BocHN-(CH2)m-X-(CH2) c R = H, alkyl, aryl ZNH
O
HO R O~O
BocHN-(CH2)m-X-(CH2) OH ---~ BocHN-(CH2)m-X-(CH2) R
ZNH ZNH
(a) LiAIH4, THF, 0 'C - 20 °C; (b) RCH=PPh3, THF; (c) Os04, NMMO, acetone-H20;
(d) CI2C=O, pyridine.
Scheme 23 R O
BocHN-(CH2)m-X-(CH2) ~ BocHN-(CH2)m-X-(CH2) R
ZNH ZNH
R = H, alkyl, aryl HO R
b BocHN-(CH2)m-X-(CH2) XR"
ZNH
X = OorS
(a) mcpba, DCM, 20 °C; (b) NaSR" or NaOR", toluene-THF, Lewis acid.

WO 95/24382 21 ~ 4 6 91 pCT/US95/02669 Scheme 24 HO R F R
BocHN-(CH2)m-X-(CH2) OH a-=b~ BocHN-(CH2)m-X-(CH2) OTr ZNH ZNH
ne 20 °C; (b) Et2NSF3, dioxane.
Scheme 25 BocHN-(CH2)m-X-(CH2) COR a BocHN-(CH2)m-X-(CH2) OH
ZNH ZNH
R = H, alkyl, aryl (a) CF31, Zn, DMF, -20'C.
Scheme 26 HO
BocHN-(CH2)m-X-(CH2) CHO gocHN-(CH2)m-X-(CH2) CN
ZNR~ ZNR~
HO
b H2N-(CH2)m-X-(CH2) C02R
R = H, alkyl ZNR~
(a)KCN, NaHS04; (b) ROH, HCI or H20, H*.

W0.95/24382 218 4 6 9 i PCT/L1S95/02669 A
Scheme 27 NH HO
NH(CH2)m-X-(CH2) C02R a NHR~
NH O
NH(CH2)m-X-(CH2) C02R

(a) Mn02, H20.
Scheme 28 OH a.b ~ ~ OMe c,d i ~ ~ --.~ 02N i -02N ; / ~ O
OMe OMe ---f H2N
02N ! / NH O / ~NH O
Boc BoC
(a) (COCI)2, TEA, DMSO, DCM; (b) (carbomethoxymethyl)triphenylphosphonium bromide; (c) ammonium chloride; (d) di-t-butyl dicarbonate; (e) H2, Pd/C.

Without further elaboration, it is believed that one skilled in the art can, using the preceding description, utilize the present invention to its fullest extent. Therefore the following preferred specific embodiments are to be construed as merely illustrative and not limitative of the remainder of the disclosure in any way whatsoever.
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 either neutral, or acidic, or basic aqueous solutions and organic solvent. The aqueous layer was extracted n times (x) with the indicated organic solvent.
1 5 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 et al: (Still, W.C.;
2 0 Kahn, M.; Mitra, A. Rapid Chromatograhic Technique for Preparative Separation with Moderate Resolution. J. 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
HCl in dioxane. Thin layer chromatograms were run on 0.25 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
3 0 spectra were obtained from either General Electric QE-300 or Varian VXR 400 MHz spectrometer with tetramethylsilane as an internal standard. 13C NMR were obtained from a Varian spectrometer at 125.8 MHz with tetramethylsilane as an internal standard.
3 5 - Example 1 WO 95/24382 2 ~ g 4 6 91 PCT/US95/02669 3S-amino-7-[(1-iminoethyl)amino]heptanoic acid NH
~N

H

O
ZHN NHBoc la 1 0 la . Boc-L-Lys ( Z ) -OH in 50 mL THF ( 3 . 8 g, 10 mmol ) was reacted with isobutyl chloroformate (1.4 mL, (10 mmol) in the presence of NMM (1.1 mL, 10 mmol). The salt was filtered and the mixed anhydride was reacted with 25 mmol diazomethane in 100 mL Et20 for 12 h. Solvent was evaporated to give an oil. This structure and subsequent structures were characterized by 1H NMR.
ZHN ~~riBoc 1b 2 0 1b. la dissolved in 50 mL EtOH was treated with Ag benzoate (0.5 g) in the presence of TEA (5 mL) for 2 h.
After filtration, the (3-amino acid ester was purified by column chromatography to give 0.84 g of product.

WO 95/24382 218 4 6 91 pCT/US95/02669 H2N mriBoc lc lc. 1b (0.84 g 12 mmol) dissolved in 30 mL MeOH was S reduced in the presence of 1 g ammonium formate and 0.2 g Pd black for 60 min. After filtration and evaporation, product was recovered.
Et NH
' 'N

H
1d Ethyl 3S-amino-7-[(1-iminoethyl)amino]heptanoate 1d. lc in 10 mL DMF was treated with methyl acetimidate (0.692 g, 6 mmol) and N,N-diisopropylethylamine (1.05 mL, 6 mmol) overnight. Solvent was removed in vacvo and the residue treated with TFA (10 mL) for 30 min. The reaction was diluted with H20 and purified by HPLC to yield 0.16 g (35.1%) of an oil. FAB MS: MH+=230.2 1. 1d (0.12 g, 0.52 mmol) dissolved in 20 mL 2N HC1 was refluxed for 60 min. The reaction was diluted with H20 and lyophilized to yield 0.107 g (1000) of an oil. FAB MS:
2 5 MH+=202.3 Example 2 WO 95/24382 218 4 b 91 PCT/US95/02669 3S-amino-6-[(1-iminoethyl)amino]hexanoic acid O
NH O
_N v H

H
2. Example 2 was prepared in the same manner as described for example 1 starting with Boc-Orn(Z)-OH (3.6 g, mmol) to yield 0.123 g (330) of an oil. FAB MS:
1 0 MH+=188.0 Example 3 N-(5S-amino-6,7-dihydroxyheptyl)ethanimidamide, dihydrochloride OH OH
NH ~ ~~~~~OH NH OH

The absolute stereochemistry of the hydroxyl group at position C-6 has not been determined. The diastereomers have been separated as described below A difference in the biological activity between the diastereomers is seen.

N-OC.-Boc-N-E-Z-L-Lys-OMe (3a) WO 95/24382 218 4 b 91 p~/US95/02669 3a. To a stirring solution of cesium carbonate (32.6 g, 0.10 mol) in 150 mL DMF was added N-oc-Boc-N-E-Z-Lys (68.3 g, 0.18 mol). After 10 min, iodomethane (51.1g, 0.36 mol) was added. After 18 h, solvent was removed in vacuo. The 5 resultant gum was washed with hexane and the hexane was decanted. The product was dissolved in 100 mL of DCM and filtered through a 100 x 70,mm pad of EM silica gel. The silica was washed with 900 mL DCM and 300 mL EtOAc which were combined. The solvent was removed in vacuo to yield 10 66.4 g (94 0) of product.
-O O H
ZHN NHBoC ZHN NHBoC
3b 3c 3b, c. To a stirring solution of 3a (7.9 g, 20 mmol) in 100 mL dry toluene cooled to -70'C was added dropwise over 10 15 min 1M DIBAL-H in toluene (40 mL, 40 mmol). After stirring an additional 20 min, the reaction was quenched with 4 mL
MeOH. ~lpon removal of the ice bath, 150 mL of saturated solution of Rochelle salt was added to the reaction. After stirring for 1 h, the layers were separated. The aqueous 2 0 layer was extracted with 2x 150 mL EtOAc. The combined organic layers were washed with 2x 200 mL H20, dried, filtered, and concentrated in vacuo. The residue was purified by flash chromatography according to Still et al.
to yield 5 . 37g (74 0 ) of 3b and 0 . 70 g ( 10 0 ) of 3c . Both 2 5 3b and 3c were white solids.
ZHN NHBoc 3d 3d. To a stirring suspension of methyltriphenylphosphonium bromide (2.18 g, 6.1 mmol) in 50 mL of Et20 was added dropwise 0.5 M potassium hexamethyldisilazide in toluene (12.2 mL, 6.1 mmol). After stirring for 1.5 h, 3b (2.22 g, 6.1 mmol) in 50 mL of Et20 was added. After 16 h, a white solid was filtered from the reaction. The filtrate was concentrated in vacuo. The residue was purified by flash chromatography to yield 1.21 g (50~) of 3d, a clear colorless gum. Anal calcd for 1 0 C2pH30N204'0.2 H20: C, 65.62; H, 8.37; N, 7.65. Found: C, 65.65; H, 8.07; N, 7.59.
HO
~O H
ZHN NHBoc 3e 3e. To a stirring solution of 3d (1.20 g, 3.3 mmol) in 80 mL of acetone:H20 (3:1) was added 4-methylmorpholine N-oxide ( 0 . 64 g, 4 . 8 mmol ) and 2 . 5 ~ Os04 in t-BuOH (3 . 4 niL, 3.4 mmol) . After 18 h, 120 mL of H20, 8 g of celiteTM, and 2 0 1.6 g Na2S204 were added to the reaction. The reaction was filtered through a pad of wet celiteTM. To the filtrate was added 200 mL of 1M KHS04.. The filtrate was extracted with 3x 200 mL EtOAc. The combined organic layers were dried, filtered, and stripped. The residue was purified by flash 2 5 chromatography to yield 0.93 g (71 ~) of 3e. Anal calcd fox C2pH32N206'0.25 H20: C, 59.91; H, 8.1?; N, 6.99. Found:
C, 59.75; H, 8.42; N, 6.77.
H
~O H
H2N NHBoc 3 0 (3f) 3f. Benzyloxycarbonyl protecting group was removed from 3e (1.38 g, 3.5 mmol) by catalytic hydrogenation using Pd black as the catalyst yielding 3f quantitatively.

3A, 3B. To a stirring solution of 3f (3.90 g, 14.9 mmol) and TEA (3.3 mL, 24 mmol) in 10 mL of DMF was added 1 0 methyl acetimidate (2.44 g, 22.2 mmol) . After 16 h, TEA~HC1 was filtered from the reaction and washed with a minimum of DMF . The filtrate was adjusted to pH 3 with 1N HC1 . The filtrate was concentrated under high vacuum. The residue was applied to a reverse phase column (YMC AQ-363-lOP, ODS) using a gradient of 20 o CH3CN/0.025 % HOAc to 50 0 CH3CN/0.025 o HOAc. The two diastereomers were separated.
The first eluting isomer was treated with 1N HC1 for 1 h at ambient temperature. The aqueous solution was lyophilized.
The yield was 0.51 g of 3A. The second eluting isomer was 2 0 treated in the same fashion to yield 0 . 40 g of 3B . Anal calcd for (3B) CgH21N302~1.75HC1~0.75 H20: C, 38.52; H, 8.71; N,14.97. Found: C, 38.60; H, 8.73; N, 13.34 .
Example 4 N1-(1-iminoethyl)-1,4-pentanediamine, dihydrochloride H NH
H~ N N
H

WO 95/24382 Z 1 g 4 6 91 PCT/US95/02669 ~O
Boc 4a 4a. A solution of 5-methyl-2-pyrrolidinone (50 g, 0.5 mol) , di-t-butyl dicarbonate ( 165 g, 0 . 7 6 mol) , DMAP ( 62 g, 0.5 mol) and Et3N (250 mL) in CH2C12 (250 mL) was stirred at room temperature for 24 h. The solvent was concentrated in vacuo and the resulting oily red solid suspended in Et20 and filtered. The Et20 solution was passed through a pad of silica gel. The solvent was removed to yield an orange liquid. The product was chromatographed to yield 82 g (83%) of a yellow liquid.
H O
Boc' N O H
4 b 4b. Sodium hydroxide (2.24 g, 56 mmol) was added to a stirring solution of 4a (4.0 g, 20 mmol) in THF:H20 (175 mL:75 mL). The resulting solution was stirred for 2 h. The 2 0 solvent was concentrated in vacuo to 75 mL. The solution was acidified with citric acid (1 M, 75 mL), extracted with EtOAc (200 mL) , dried, and concentrated in vacuo to yield 5.36 g of an oil. The product was crystallized from Et20/hexane to yield 4.24 g (98o)of a white solid.
H O
Boc' N NH2 4c 4c. To a stirred solution of 4b (4.0 g, 18 mmol) and TEA
3 0 (2.6 mL, 18 mmol) in THF (50 mL) at -10°C was added isobutyl WO 95/24382 218 4 6 91 PCT~S95102669 chloroformate (2.39 mL, 18 mmol) dropwise and the solution stirred for 20 min . Ammonium hydroxide (3 . 9 mL, 28 0 ) was added and the resulting solution was stirred for 18 h allowing to warm to room temperature. The solution was concentrated in vacuo and the residue suspended in boiling EtOAc (80 mL) and filtered. This was repeated. The filtrate was concentrated to 30 mL and the solid collected to yield 3.82 g.
H
Boc'N ~N
4d 4d. To a stirred solution of 4c (3.5 g, 16 mmol) in THF
(20 mL) at 0°C was added TFAA (2.5 mL, 17.5 mmol) dropwise and the solution stirred for 20 min. The solution was poured onto Et20 (125 mL) and NaHC03 (satd, 25 mL), the layers separated and the organic solution extracted with NaHC03 ~ (satd) and brine (satd. ) , dried, filtered, and 2 0 concentrated in vacuo to yield 2.88 g of an oil. The product was vacuum distilled (bp 130°C @ 0.6 mmHg) to yield 2.2 g (690) of a yellow liquid.
H
Boc' N NH2 4e 4e. A solution of 4d (1.7 g, 8.6 mmol) in EtOH was treated with H2 (300 psi) over Raney nickel at 50°C for 8 h.
3 0 The reaction mixture was filtered and concentrated in vacuo to yield 1.25 g (730) of a colorless oil.

H NH
Boc'N N' H
4f 5 4f. A solution of 4e (1.0 g, 4.9 mmol) and ethyl acetimidate hydrochloride (0.62 g, 5 mmol) in anhyd. EtOH
(25 mL) was stirred for 18 h. The reaction solution was concentrated in vacuo to yield 1.46 g of a white foam. This material was used in the next step without further 10 purification.
H NH
H~ N N
i H

15 4. A solution of 4f (1.46 g, 4.9 mmol) in acetone (25 mL) was treated with HCl (10 mL, 2 M in MeOH) , and stirred for 10 min. The reaction mixture was concentrated in vacuo and triturated with ethanol and THF to yield an oil.
Crystallization of the oil was attempted from i-propanol.
2 0 The solution was concentrated in vacuo to obtain a foam 0.42 g (390) which was dried. Anal. Calcd for C~H1~N3 - 2 HC1 -0.15 H20 - 0.15 i-PrOH: C, 39.27; H, 9.07; N, 18.44; C1, 31.12. Found: C, 39.25; H, 9.53; N, 18.04; C1, 31.52.
2 5 Example 5 N1-(1-iminoethyl)-1,5-heptanediamine H H
H, N N
jN[ H

WO 95/24382 ~ ~ 8 4 6 91 PCT/US95/02669 H H
BoC' N N~H
5a 5a. A solution of 3d (500 mg, 1.38 mmol) in AcOH/EtOH
was treated with H2 (5 psi) over Pd black for 21 h. The reaction mixture was filtered and concentrated in vacuo .
The residue was dissolved in CH2C12(125 mL) and extracted with NaOH (1 M), dried over Na2SOq, filtered, and concentrated in vacuo to yield 0.32 g of a white gum.
H H
Boc' N N
NH
5b 5b. A solution of 5a (0.29 g, 1.26 mmol) and ethyl acetimidate hydrochloride (0.156 g, 1.3 mmol) in EtOH (15 mL) was stirred for 18 h. The reaction solution was 2 0 concentrated in vacuo to yield 0.40 g of a white gum. This material was used in the next step without further purification.

5. To a stirred solution of 5b (0.40 g, 1.26 mmol) in AcOH (glacial, 10 mL) was added HC1 ( 6.95 M in dioxane, 14 mmol). The resulting solution was stirred for 2 h. The solution was concentrated in vacuo to yield 0.41 g of a gum.
3 0 This material was purified by reversed phase HPLC on a C-18 support (7:3 CH3CN:H20) to yield 95 mg of clean product as a glass. HRMS calcd for CgH22N3: 172.1814. Found: 172.1809.
Example 6 N1-(1-iminoethyl)-5-phenyl-1,5-pentanediamine H
~N

N O
Boc 6a 6a. The reaction for example 4a was repeated on a 0.4 mol scale with valerolactam. The yield of the reaction was quantitative.
H
Boc' N
O
6b 6b. A solution of 6a (5 g, 25 mmol) in THF (125 mL) at -78°C was treated with phenylmagnesium bromide (9.5 mL, 3.0 M). The resulting solution was stirred at -72°C for 35 min 2 5 then poured onto brine (satd) and extracted with Et20. The organic solution was dried, filtered, and concentrated in va cuo to yield 6.42 g of an oil. The product was chromatographed and recrystallized from hexane to yield 3.78 W0.95/24382 218 4 6 91 PCT~S95/02669 g (550) of a white solid. Anal. Calcd for C16H23N03: C, 69.28; H, 8.36; N, 5.05. Found: C, 69.30; H, 8.84; N, 4.95.
H /
Boc' N
$ NOH
6c 6c. A stirred suspension of 6b (0.50 g, 1.8 mmol) in EtOH (3 mL) was treated with a solution of hydroxylamine hydrochloride (0.25 g, 3.6 mmol), sodium acetate (0.25 g, 3.8 mmol) in H20 (3 mL) . The solution was re fluxed for 4.5 h, during which time a solution formed. After cooling to room temperature H20 (50 mL) was added and the mixture extracted with CHC13 (3 x 30 mL). The CHC13 extracts were combined, dried, filtered, and concentrated in vacuo to yield 420 mg (80$) of a white solid. Anal. Calcd for C16H24N203~ C, 65.73; H, 8.27; N, 9.50. Found: C, 65.79; H, 8.79; N, 9.53.
H /
Boc' N
2 0 ~ NH2 6d 6d. A solution of 6c (3.54 g, 24 mmol) in EtOH was treated with H2 (5 psi) over loo Pd/C for 24 h. The reaction 2 5 mixture was filtered and concentrated in vacvo to yield 2.61 g (77 0) of a colorless oil. Anal. Calcd for C16H26N202 : ' 0.4 EtOH: C, 67.98; H, 9.64; N, 9.44. Found: C, 68.19; H, 9.38; N, 9.11.

WO 95/24382 21$ 4 6 91 PCT/US95/02669 H
Boc' N
H'N~Cbz 6e 6e. To a solution of 6d (2.1 g, 7.54 mmol) in EtOAc (100 mL) in a separatory funnel was added NaOH (1 M, 60 mL) and benzylchloroformate (1.93 g, 11.31 mmol). The mixture was shaken for several minutes and the layers separated, The EtOAc solution was extracted with brine, dried, filtered, and concentrated in vacuo to yield an oil. The oil was chromatographed to yield 2.34 g (750) of a white solid.
H
H~N ~ I
H'N'Cbz 6f 6f. A solution of 6e (2.0 g, 4.85 mmol) in CH2C12 (25 mL) at 0'C was treated with TFA (20 mL) and allowed to warm to room temperature over 1.5 h. The solution was concentrated in vacuo to yield a yellow oil. The oil was dissolved in CHC13 and extracted with NaOH (1 M) and brine (satd) , dried over Na2S04, filtered, and concentrated in vacuo to yield 1.35 g (890) of a gum.
H
1f N w I
N H H~N'Cbz 6g 6g. A solution of 6f (1.3 g, 4.16 mmol) and ethyl acetimidate hydrochloride (0.533 g, 4.2 mmol) in EtOH (20 WO 95/24382 218 4 6 91 pCT~S95/02669 mL) was stirred for 18 h. The reaction solution was concentrated in vacuo to yield 1.65 g of a white foam. This material was purified by reversed phase HPLC to yield 1.09 g of a foam. Anal. Calcd for C21H2~N3O2'1HC1 ~ 0.75 H20: C, 5 62.52; H, 7.37; N,10.42. Found: C, 62.82; H, 7.05; N, 10.12.

10 6. A solution of 6g (0.94 g, 2.41 mmol) in AcOH was treated with H2 (5 psi) over Pd black for 20 h. The reaction mixture was filtered and concentrated in vacuo . The residue was dissolved in EtOH (10 mL) and HC1/Dioxane (1 mL, 5.8 M) added and concentrated in vacuo to yield 0.52 g 15 (740) of a white powder. Anal. Calcd for C13H21N202'2 HC1~0.75 H20 . 0.2 EtOH: C, 51.10; H, 8.22; N,13.34; Cl, 22.51. Found: C, 50.98; H, 7.82; N, 13.66; C1, 22.20.
Example 7 2 0 N-[5-amino-5-(2-hydroxyphenyl)pentyl]ethanimidamide H
'' N
~NH NH2 OH

H
Boc' N
O OTHP
7a 7a. The reaction for example 6b was repeated on a 25 3 0 mmol scale with 2-(tetrahydropyran-2-yloxy)phenyllithium.
The 2-(tetrahydropyran-2-yloxy)phenyllithium was prepared from 2-(tetrahydropyran-2-yloxy)phenyl bromide and n-BuLi in THF at -78°C. The crude product was chromatographed to yield 4.07 g (43~) of a yellow oil.
7. Example 7 is prepared in the same manner as described for example 6.
Example 8 N-[5-amino-5-(4-hydroxyphenyl)pentyl]ethanimidamide H / OH
''N
~NH NH2 8. Example 8 is prepared in the same manner as described for example 6 starting with 4-(tetrahydropyran-2-yloxy)phenyllithium.
2 0 Example 9 N-(5-aminononyl)ethanimidamide H
'' N
~NH NH2 9. Example 9 is prepared in the same manner as described for example 6 starting with n-butyllithium.
Example 10 ~3-amino-4-[(1-iminoethyl)amino]benzenepropanoic acid, dihydrochloride hydrate H
~N
NH I / OH
H.N,H O

O
- O' N + ( ~
/ OH

10a 10 10a. A mixture of 4-nitrobenzaldehyde (39 g, 0.26 mol), malonic acid (30.5 g, 0.29 mol) and ammonium acetate (49 g, 0.64 mol) in AcOH was heated at 100°C for 5 h, followed by the addition of HCl (25~, 200 mL) and continued heating at 100°C for 5 additional hours. The reaction mixture was cooled to room temperature and H20 (300 mL) added, the resulting precipitate was filtered and washed with H20 (100 mL) . The filtrate and wash were combined and concentrated in vacuo, followed by the addition of H20 (300 mL) . The resulting mixture was heated on a steam bath, decolorized 2 0 with carbon and filtered through celiteTM° The pH of the solution was adjusted to 7 with NH40H (cone.) and the resulting precipitate collected. The solid was washed with H20 (100 mL), methanol:H20 (1:1, 100 mL), methanol:Et20 (1:1, 100 mL) and Et20 (200 mL). The solid was dried in 2 S vacuo to yield 29.8 g of a yellow solid:

O
"+
_ 0.N I ~
/ OH
H, N. B00 lOb 10b. A solution of 10a (5.0 g, 24 mmol), di-t-butyl dicarbonate (5.7 g, 26 mmol) in NaOH (1 M, 50 mL) and dioxane (50 mL) was stirred for 4 h. The solvent was concentrated to 50 mL to which was added EtOAc (400 mL) and KHS04 (1M, 75 mL). The layers were separated and the organic layer was washed with brine (satd.), dried, filtered, and concentrated in vacuo to yield 8.5 g of a yellow foam.
O
"+
_ O.N I

H'N' BocO
lOc 10c. Example lOc was prepared in the same manner as 3a.
H
H'N

H, N. B00 lOd 10d. A solution of lOc ( 8 mmol) in EtOH was treated with H2 (5 psi) over loo Pd/C for 18 h. The reaction mixture was filtered and concentrated in vacuo to yield the product.

H
N
N H ( ~ OCH3 H' N' Boc~
10e 10e. The reaction for example 6g was repeated using lOd on a 5 mmol scale.
H
''N
NCH ~ ~ OCH3 H.N.H O
lOf 10f. A solution of 10e (890 mg, 2.4 mmol) in CH2C12 . TFA
(1:1, 50 mL) was stirred at 0°C for 15 min. The solvent was removed in vacuo and the residue was dissolved in water (100 mL) and the extracted with EA. The pH of the aqueous solution was adjusted to 11 with K2C03 and extracted with CH2C12. The CH2C12 extracts were dried (Na2S04) and concentrated. The residue was chromatographed on silica gel (9:1:1; ACN:H20:AcOH) to to yield lOf 50 mg (5 0) as a foam. Anal. Calcd for C12H1~N302~3 AcOH~1 H20: C, 49.87; H, 7.21; N,9.69. Found: C, 49.61; H, 6.96; N, 9.78. HRMS
2 0 calcd.: 235.1320. Found: 235.1320.
10. A solution of lOf (20 mg, .5 mmol) in HC1 (2N, 5 mL) 2 $ was refluxed for 1 h. The solvent was removed via lyophilization to yield 10 24 mg as a foam. Anal. Calcd for C11H15N302'2.3 HC1~0.6 H20: C, 41.82; H, 5.90; N,13.30.

Example 11 0c- [1-amino-5- [ (1 iminoethyl)amino]pentyl]benzenemethanol hydrochloride dehydrate NH NH2 _ ~N
I "'' H OH

1 O N-OC-Z-N-~-Boc-L-Lys-N (OMe) Me (11a) 11a. To a stirring solution of N-OC-Z-~-Boc-L-Lys (5 g, 13.8 mmol), N,O-dimethylhydroxylamine HCl (3.9 g, 39.5 mmol),1-hydroxybenzotriazole hydrate (2 g, 14.5 mmol), and 15 triethylamine (13.2 g, 17 mL, 130 mmol) in 75 mL of dimethylformamide (DMF) cooled in an ice bath was added EDC
(2.8 g, 14.5 mmol). After stirring 55 h at ambient temperature, triethylamine hydrochloride was filtered from the reaction mixture and the filtrate was concentrated in 2 0 vacuum. The residue was distributed between 150 mL of ethyl acetate (EtOAc) and 75 mL of 1M KHS04 solution. The layers were separated. The organic layer was washed with 1x 75 mL of saturated KHC03 solution and 1x 75 mL of brine and was worked up in~the usual manner giving 5.3 g of lla 25 (950) .
11b. To a stirring solution of lla (1.8 g, 4.26 mmol) and N,N,N,N-tetramethylethylendiamine (1.63 g, 2.12 mL, 14.06 mmol) in 50 mL of dry THF at -72~C was added phenyllithium, 3 0 1.8 M solution in cyclohexane, (1.18 g,7.8 mL, 14.06 mmol).
After stirring at the same temperature for 2.5 h, the reaction mixture was added to 50 mL of 1M KHSOq solution and WO 95124382 218 4 6 91 P~'~S95/02669 50 mL of EtOAc. The layers were separated, the organic layer was washed with 1x 30 mL of brine and worked up in the usual manner giving 2.8 g of crude product which was purified using column chromatography. The yield of llb was S 1.3 g (69.5%) .
11c. To example llb (1.3 g, 2.95 mmol) dissolved in 30 mL
of acetic acid was added 3 mL of 5N HC1/dioxane. The reaction was stirred for 20 min. at ambient temperature concentrated under vacuum. The residue was dried, treated with Et20, washed with hexane, and dried yielding 1.0 g (90.90) of 11c. Anal.calcd. for C2pH24N203-HC1-0.4 H20: C, 62.54; H, 6.77 N, 7.29 CL, 9.23. Found: C, 62.88 H, 6.80; N, 7.22; C1, 9.18.
11d. 0.5 g of llc dissolved in 10 mL of water was neutralized with Na2C03 (pH-9-10), the oil was extracted with 3x 15 mL of EtOAc and the organic solution was worked up in the usual manner giving 0.45 g of 11d.
11e. A solution of lld (0.45 g, 1.32 mmol) and ethyl acetimidate hydrochloride (0.2 g, 1.455 mmol) in 15 mL of ethanol was adjusted to pH 9-10 using a NaOH/ethanol solution. After stirring for 1 h at ambient temperature, 2 S the reaction was acidified to pH 2 with 5N HC1/dioxane. The reaction mixture was filtered from NaCl and concentrated in vacuum. The crude product (0.5 g) was purified using reverse phase separation, giving 0.225 g of lle (40.910).
Anal. calcd. for C22H2~N303-HCl-0.5 H20: C, 61.89; H, 6.85;
3 0 N, 9.84; C1, 8.30. Found: C, 61.68; H, 6.50; N, 9.88; C1, 8.18.
11A, 118. Example lle ( .36 g, 0. 86 mmol) was reduced under catalytic hydrogenation conditions using Pd black at 60 psi 35 H2 in 50°s EtOH/AcOH solution for 24 h. The yield of the WO 95/24382 ~ ~ 8 4 6 91 PCT/US95/02669 crude product was 0.35 g. After reverse phase separation two products were isolated: the faster running isomer (11A) 0.085 g and the slower running isomer 0.1 g (11B). Faster running isomer analysis: calcd. for C14H23N30. 1.5HC1, S 0.4AcOH, 2H20: C, 48.82; H, 8.33; N, 11.54. Found: C, 48.56; H, 7.79; N, 11.95.
Example 12 N-[SS-amino-6-oxo-6-(2-thienyl)hexyl]ethanimidamide, 1 0 hydrate ~N y H O

1 5 12a . 12a was prepared on a 2 . 84 mmol scale in the same manner as described for llb using lla and 2-thiophenelithium to yield 0.6 g (47.20) of 12a after chromatography. Anal calcd. for: C23H3pN2O5S: C, 61.86; H, 6.77; N, 6.27. Found: C, 61.53; H, 6.91; N, 6.12.
12b. 12b was prepared from 12a (0.6 g, 1.34 mmol) in the same manner as for llc yielding 0.4 g (85.10).
12c. 12c was prepared from 12b (0.4 g, 1.15 mmol) in 2 5 the same manner as for lle to yield 0.44 g of crude product.
12. To a solution of 12 c (0.44 g, 1.14 mmol) and thioanisole (0.51 g, 0.44 mL, 2.28 mmol) in 10 mL of TFA at 3 0 0'C trimethylsilyl trifluoromethanesulfonate (TMSOTf) (0.28 g, 0.27 mL, 2.28 mmol) is added. After mixing at same W0.95124382 218 4 6 91 PCT/US95/02669 temperature for 1 h, Et20 is added. The crude 12 is filtered and is washed with Et20.
Example 13 N-[5S-amino-6-hydroxy-6-(tetrahydrofuran-2-yl)hexyl]ethanimidamide ~N
I _ H OH

1 3. Example 13 was prepared in the same manner as example 11 on a 9.2 mmol scale starting with 2-bromofuran.
Example 14 N-(5S-amino-6-oxoheptyl)ethanimidamide, dihydrochloride ~N
I
H O

14a. To a stirring solution of lla (1.0 g, 2.4 mmol) and N,N,N,N-tetramethylethylendiamine (0.96 g, 1.25 mL, 8.3 mmol) in 30 mL of dry THF at the -72°C was added methyllithium, 1.4 M solution in Et20, (5.9 mL, 8.3 mmol) .
2 5 After stirring at same temperature for 3 h, the reaction mixture was added to 50 mL of 1M KHS04 solution and 50 mL of EtOAc at 0 °C. The layers were separated, the organic layer was washed with lx 30 mL of brine and worked up in the usual manner giving 2.8 g of crude product which was purified WO 95/24382 218 4 6 91 P~'~595/02669 using column chromatography. The yield of 14a was 0.9 g (550) .
14b. To example 14a (0.5 g, 1.2 mmol) in 10 mL of acetic acid was added 2 mL of 6N HC1/dioxane. The.reaction was stirred for 20 min at ambient temperature then concentrated under vacuum. The residue was dissolved in H20 and lyophilized yielding 0.4 g (1050) of 14b.
14c. To a solution of 14b (0.4 g, 1.2 mmol) and TEA
(0.56 mL, 3.9 mmol) in 10 mL of DMF was added methyl acetimidate hydrochloride (0.43 g, 3.9 mmol). After stirring for 16 h at ambient temperature, the reaction was filtered. The filtrate was concentrated under vacuum. The reaction mixture was partitioned between 15 mL 1N HC1 and 20 mL DCM. The crude product from the aqueous HC1 after stripping was purified using reverse phase separation, giving 0.26 g of 14c (60.50).
2 0 14. 14c (0.26 g, 0.73 mmol) was reduced under catalytic hydrogenation conditions using Pd/C at 5 psi H2 in 500 MeOH/HC1 solution for 3 h. The yield of product was 0.18 g (94.70). Analysis calcd. for CgH1gN30-2 HC1-H20: C, 39.14;
H, 8.39; N, 15.21. Found: C, 39.24; H, 8.32; N, 14.99.
Example 15 N-(5S-amino-6,7-dihydroxy-6 methylheptyl)ethanimidamide, hydrochloride dihydrate ~N OH ~N OH
~OH H HO ..I'~.

The absolute stereochemistry of the hydroxyl group at position C-6 has not been determined. The diastereomers have been separated as described below A difference in the biological activity between the diastereomers is seen.
15a. To a stirring suspension of methyltriphenylphosphonium bromide (6.21 g, 17.4 mmol) in 5 150 mL of toluene was added dropwise 0.5 M potassium hexamethyldisilazide in toluene (35.6 mL, 17.4 mmol). After stirring for 1.5 h, 14a (6.85 g, 17.4 mmol) in 50 mL of toluene was added to the stirring suspension cooled to -20 °C. After 5 h, the reaction was warmed to 0 'C, washed 2x 10 100 mL of 1M KHS04, lx 100 mL of brine, dried, filtered, and concentrated in vacuo.. The residue was purified by flash chromatography to yield 5.3g (80~) of 15a, a white solid.
15b. To a stirring solution of 15a (3.3 g, 8.8 mmolj I5 in 150 mL of acetone:H20 (3:1) was added N-methylmorpholine N-oxide (2.05 g, 17.5 mmol) and 2.5 $ OsOg in t-BuOH (9.5 mL, 0.9 mmol). After 18 h, 100 mL of H20, 25 g of celiteTM, and 6 g NazS204 were added to the reaction. The reaction was filtered through a pad of wet celiteTM. To the filtrate 2 0 was added 180 mL of 1M KHS04 . The filtrate was extracted with 3x 250 mL EtOAc. The combined organic layers were dried, filtered, and stripped. The residue was purified by flash chromatography to yield 3.5 g (97 ~) of 15b.
2 5 15c. To a stirring solution of 15b (1.6 g, 3.9 mmol) in mL of HOAc was added 2.5 mL of 4N HC1/dioxane. After 30 min, solvent was removed under vacuum to quantitatively recover 15e.

WO 95/24382 218 4 b 91 PCT/US95/02669 15d. Example 15d was prepared in the same manner as described in example 14c starting with 15c (1.36 g, 3.9 mmol) . The residue was applied to a reverse phase column (YMC AQ-363-10P, ODS) using a gradient of CH3CN/0.025 0 HOAc. The first eluting isomer,l5d-1, weighed 0.11 g; the second eluting isomer, 15d-2 weighed 0.28 g; a mixture of the two weighed 0.18 g.
15A. Example 15A was prepared in the same manner as described for example 14 starting with 15d-1 (1.1 g, 2.6 mmol). After lyophilization, 0.82 g of 15A was recovered.
Analysis calcd. for C1pH23N3O2-2 HCl-1.75 H20: C, 37.33; H, 8.93; N, 13.06. Found: C, 37.25; H, 8.70; N, 12.95.
158. Example 15B was prepared in the same manner as described for example 14 starting with 15d-2 (0.28 g, mmol). After lyophilization, 0.21 g of 15B was recovered.
Analysis calcd. for C1pH23N302-2 HC1-2.2 H20: C, 36.41; H, 8.98; N, 12.74. Found: C, 36.31; H, 8.97; N, 12.34.
Example 16 N-(5S-amino-6,7-dihydroxyoctyl)ethanimidamide, dihydrochloride hydrate ~N ' _N
I I
H OH H OH
16A, 16B
16A,16B. Example 16A and 16B were prepared in the same manner as described for examples 3A and 3B starting with 3b and ethyltriphenylphosphonium bromide. 16B: Analysis calcd. for C1pH23N302-2 HC1-1.8 H20: C, 37.22; H, 8.93; N, 3 0 13.02. Found: C, 37.47; H, 9.05; N, 12.93.

. CA 02184691 2004-05-26 Example 17 4S-amino-2,3-dihydroxy-8-[(1-iminoethyl)amino]octanoic acid S

'' 'N COOH
i 17a. Example 17a is prepared starting with 3b and (carbomethoxymethyl)triphenylphosphonium bromide.
17b. To a stirring solution of 17a (3.3 mmol) in 80 mL of acetone:H20 (3:1) is added N-methylmorpholine N-oxide (0.64 g, 4.8 mmol) and 2.5 a OsOq in t-BuOH (3.4 mL, 0.34 mmol) .
After 18 h, 120 mL of H20, 8 g of celiteTM, and 1.6 g Na2S204 are added to the reaction. The reaction is filtered through a pad of wet celiteTM. To the filtrate is added 200 mL of 1M
KHS04. The filtrate is extracted with 3x 200 mL EtOAc. The combined organic layers are dried, filtered, and stripped.
2 0 17c. Benzyloxycarbonyl protecting group is removed from 17b by catalytic hydrogenation using Pd black as the catalyst yielding 17c quantitatively.
17d. To a stirring solution of 17c (14.9 mmol) and TEA
2 5 (3.3 mL, 24 mmol) in 10 mL of DMF is added methyl acetimidate (2.44 g, 22.2 mmol). After 16 h, TEA.HCl is filtered from the reaction and is washed with a minimum of DMF. The filtrate is adjusted to pH 3 with 1N HCl. The filtrate is concentrated under high vacuum. The residue is 3 0 applied to a reverse phase column (YMC AQ-363-10P, ODS) using a gradient of 20 ~ CH3CN/0.025 ~ HOAc to 50 CH3CN/0.025 ~ HOAc.

WO 95/24382 Z ~ g ~ 6 91 PCT/US95/02669 17e Example 17d is treated with 1N HC1 for 1 h at ambient temperature. The aqueous solution is lyophilized to give 17e .
S
17. 17d dissolved in 20 mL 2N HC1 is refluxed for 60 min. The reaction is diluted with H20 and lyophilized.
Example 18 N-(6,7-diacetyloxy-5S-aminoheptyl)ethanimidamide, hydrochloride monohydrate N OAc I
H OAc 1 S 18a To a stirring solution of 3e (0. 90 g, 2.3 mmol) and DMAP (0.61 g, 5.0 mmol) in DCM was added acetic anhydride (2.lmL, 23 mmol). After 18 h, solvent was removed under vacuum., The residue was taken up in 50 mL EtOAc which was washed with 3x 50 mL satd KHC03 solution, 1x 50 mL 1M KHS04, 2 0 and lx 50 mL H20. The organic layer was dried over Na2S04 anhydrous, filtered, and stxipped to yield 0.99 g (89a) of 18a, a pale yellow glass. Anal calcd for C24H36N208~0.2 H20: C, 59.54; H, 7.58; N, 5.79. Found: C, 59.75; H, 8.42; N, 6.77.

18b Example 18b (0.90 g, 1.9 mmol) was prepared in same manner as 3f to yield 0.64 g (1.8 mmol) of 18b .
18c. To a stirring solution of 18b (0.64 g, 1.8 mmol) in 3 0 10 mL of DMF was added a 2 mL solution of methyl acetimidate (0.10 g, 0.9 mmol) which had been neutralized with TEA (0.12 mL, 0.9 mmol) and filtered through glass wool to remove WO 95/24382 21 ~3 4 6 91 pCT~s95/02669 TEA~HC1. This was repeated 4x over t:.o hours. After stirring an additional 2 h, the reaction was adjusted to pH
3 with 1N HC1. After solvent was removed under vacuum, the crude product was purified by reverse phase chromatography.
Not only was desired product 18c (0.38 g, 510) obtained but also the monoacetoxy compound, 19a (0.11 g).
18. Example 18 was prepared from 18c (0.38 g, 0.9 mmol) dissolved in 2 mL HOAc to which was added 1 mL 4N
HC1/dioxane. The solvent was removed under vacuum. The residue was dissolved in H20 and lyophilized to give 18 (0.26 g, 810). Analysis calcd. for C13H25N304'1.75 HC1~1 H20: C,42.29; H, 7.85; N, 11.38. Found: C, 42.41; H, 7.57;
N, 10.68.
Example 19 N-(5S-amino-6-hydroxy-7 acetoxyheptyl)ethanimidamide, hydrochloride monohydrate N OAC
OH
19. Example 19a (0.11 g, 0.29 mmol) was dissolved in 1 mL of HOAc. (see example 18c for isolation of 19a) To the 2 5 above solution was added 1 mL of 4N HC1/dioxane . After 5 min, the solvent was removed under vacuum and the residue taken up in H20 and lyophilized. Analysis calcd. for C11H23N303'2 HC1~1.2 H20: C,38.87; H, 8.13; N, 12.36. Found:
C, 38.81; H, 8.01; N, 12.07.
Example 20 N-(5S-amino-6,7,8-trihydroxyoctyl)ethanimidamide /\N OH
I
H OH
20a. 17b dissolved in THF is treated with LiBH4 to remove 5 the benzoxycarbonyl group and reduce the ester to the alcohol.
20b To a stirring solution of 20a (14.9 mmol) and TEA
(3.3 mL, 24 mmol) in 10 mL of DMF is added methyl 10 acetimidate (2.44 g, 22.2 mmol). After 16 h, TEA-HC1 is filtered from the reaction and is washed with a minimum of DMF. The filtrate is adjusted to pH 3 with 1N HC1. The filtrate is concentrated under high vacuum. The crude product is purified by reverse phase chromatography 20 Example 20b is treated with 1N HC1 for 1 h at ambient temperature. The aqueous solution is lyophilized to give 20..
2 0 Example 21 N-(5S-amino-7,8-dihydroxyoctyl)ethanimidamide NH NHp pH
~N OH
I
H
2 5 21. Example 21 is prepared in the same manner as 3A, 3B
starting with 1b.
Example 22 N-[5S-amino-5-(4-methyl-2-oxo-1,3-dioxolan-4 30 yl)pentyl]ethanimidamide WO 95/24382 218 4 6 91 pCT/US95/02669 NH NHp ~N O
~ O' J
~H
O
22. 15b is treated with phosgene to generate cyclic carbonate. Example 22 is synthesized by methods described in example 15.
Example 23 N-(5S-amino-6-hydroxy-7-methoxyheptyl)ethanimidamide ~N OMe OH
H
23a. To a stirring solution of 3d (3.62 g, 10 mmol) in 25 mL of DCM was added m-chloroperbenzoic acid (2.59 g, 15 1 5 mmol) . . After 16 h, solvent was removed under vacuum. The resulting residue was taken up in 100 mL of EtOAc and washed with 3x 100 mL satd KHC03 solution. The organic layer was dried, filtered, and stripped. The crude product was purified by flash column chromatography to give 2.89 g (760) 2 0 of 23a .
23b. Example 23b is prepared in the manner described in Tetrahedron Lett, 1994, 35, 8977-80. To a stirring suspension of NaOMe in 15 mL of toluene-THF (2:1) cooled to 2 5 -78 °C is added Et2A1C1 (3.6 mL, 3.6 mmol [1M solution]).
After 30 min, 10 mL toluene solution of 23a (0.63 g, 1.7 mmol) is added dropwise to NaOMe suspension. The reaction is quenched after 1.5 h with Na2SOq-10 H20 (5 g) and Na2C03 (0.3g). After removing the ice bath, the suspension is WO 95/24382 2 ~ 8 4 6 91 p~~s95/02669 s2 stirred for 1 h. The salts are filtered from reaction and the filtrate is concentrated to yield 23b.
23. Example 23 is prepared from 23b in the same manner S as described in examples 3f and 3.
Example 24 N-[5S-amino-6-hydroxy-7 (ethylthio)heptyl]ethanimidamide ~N S~
OH
H
24a. To stirring ethanethiol (0.19 mL, 2.5 mmol) was added tetra-n-butylammonium fluoride. After 15 min, 23a 1 S ( 0 . 7 9 g, 2 . 1 mmol ) in 15 mL CH3CN was added . After 16 h, the solvent was removed. The crude product is purified by flash chromatography.
24b. Example 24a is treated with LiAlHq to remove the benzyloxycarbonyl protecting group.
2 0 24. Example 24 is prepared from 24b using conditions described in example 3.
Example 25 N-[5S-amino-6-hydroxy-7 2S (methylsulfinyl)heptyl]ethanimidamide ~N S/
OH p H
25. Example 25 is prepared from 24 by treatment with 3 0 30~ H202 and acetic acid at room temperature for 1 h.

WO 95/24382 2 l 8 4 6 91 PCT/US95/02669 Example 26 N-[5S-amino-6-hydroxy-7 (methylsulfonyl)heptyl]ethanimidamide ~N S/
H OH O
26. Example 26 is prepared from 24 by treatment with 30% H202 and acetic acid at 60 °C for 4 h.
Example 27 N-[5S-amino-6-hydroxy-7 [(phenylmethyl)thioJheptyl]ethanimidamide / 'N S w OH
27. Example 27 is prepared from 23a and benzyl mercaptan in the same manner as 24.
Example 28 2 0 N-[5S-amino-6-hydroxy-7-[(phenylmethyl)sulfinyl]heptyl]ethanimidamide / \N S w H OH O
2 5 28. Example 28 is prepared in the same manner as 25 starting with 27.
Example 29 WO 95/24382 2 ~ g ~ 6 9 l pCT~s95/02669 N-[5S-amino-6-hydroxy-7 [(phenylmethyl)sulfonyl]heptyl]ethanimidamide NH NHZ
/ 'N O w I OH O
H
29. Example 29 is prepared in the same manner as 26 starting with 27.
Example 30 4S-amino-2,2-difluoro-3-hydroxy-8-[(1-iminoethyl)amino]-3-methyloctanoic acid ~N COOH
I OH
H
1 S 30a . To a refluxing suspension of Zn (2 mg-atm) and ethyl bromodifluoroacetate (2 mmol) in 10 mL is added dropwise a solution of 14a (1 mmol) in 2 mL of THF. After 1 h, the reaction is cooled to room temperature. To the reaction is added 20 mL of EtOAc and 20 mL 1M KHSOq. The layers are 2 0 separated and the organic layer is treated in the normal manner to yield 30s.
30b. Conditions described in example 14 are used to prepare 30b from 30a.
30. To remove the ethyl ester from 30b, conditions 2 5 described in example 1 are used.
Example 31 N-(5S-amino-6-fluoro-7-hydroxy-6 methylheptyl)ethanimidamide ~N OH
I
H F
31a . To a stirring solution of 15b ( 1 . 5 mmol) in 10 mL
of pyridine is added Ph3CC1 (1.5 mmol) . After 16 h, the 5 reaction is concentrated under vacuum. The residue is taken up in 20 mL of EtOAc and is washed with 3x 20 mL 1M KHS04, 2x 20 mL saturated KHC03, and lx 20 mL brine. The organic layer is treated in the normal manner to obtain 31a.
31b . To a stirring solution of 31a in dioxane is added 10 Et2NSF3. After 40 h, the reaction is concentrated under vacuum and chromatographed to obtain 31b.
31. Using methodology described for example 15, example 31 is synthesized from 31b.
15 Example 32 N-[5S-amino-6,7-dihydroxy-7-(2-thienyl)heptyl]ethanimidamide, dihydrochloride ~N S
2 HC1 pH

H~NBOC
Z~ ~ S
N v v v H
32a 32. A mixture of palladium acetate (Johnson Matthey, 0.29 mmol), tri-o-tolylphosphine (0.6 mmol), 2-bromothiophene (16.0 mmol), and triethylamine (16 mmol) is refluxed under nitrogen for 30 min. The mixture is cooled to room temperature, and 3d (14.4 mmol) in 6 mL of acetonitrile is added. The reaction is refluxed for 24 h, cooled to room temperature, and stripped of all solvent under reduced pressure. The residue is partitioned between sat. NaHC03 and EtOAc and the organic phase is dried (MgS04), filtered, and stripped. The residue is chromatographed on silica gel to give 32a.
H~ NBOC OH
Z wN S
H OH
32b 32b. Osmium tetroxide is reacted with 32a by the method used in the preparation of 3e, to yield 32b.
H~ NBOC pH
H wN S
H OH
32c 32c . A solution of 32b in AcOH is treated with H2 (5 psi) over Pd black for 20 h. The reaction mixture is assessed by thin layer chromatography to find the extent of 2 5 reaction. If necessary, fresh Pd black is added and the reaction continued. This process is repeated until the reaction is completed. The reaction mixture is filtered and concentrated in vacuo to yield 32c.

WO 95/24382 218 4 6 91 pCT/US95/02669 NH H~NBOC OH
~N S
H OH
32d 32d. An equimolar solution of 32c and ethyl acetimidate hydrochloride in EtOH is stirred for 18 h.
The reaction solution is concentrated in vacuo to yield a white foam. This material is purified by reversed phase HPZC to yield 32d.
32. To a stirred solution of 32d in AcOH (glacial) is added HC1 ( 6. 95 M in dioxane) . The resulting solution is stirred for 2 h. The solution is concentrated in vacuo and triturated with diethyl ether to yield 32.
1$
Example 33 N-[5S-amino-6,7-dihydroxy-7-(1H-imidazol-5-yl)heptyl]ethanimidamide, trihydrochloride N
N v v 3 HCl ~ ~ /
N

H~NBOC
ZwN ~ N
H v v /
N
33a 33a. 4-Bromoimidazole (K&K Laboratories) is treated as described in the preparation of 32a, replacing the 2-bromothiophene in that preparation. The product is 33a.
H ~ NBOC OH
ZEN N
H v OH
N
33b 33b. By the method of Example 3e, osmium tetroxide is reacted with 33a to yield 33b.
H~ NBOC OH
H
HwN N
I v v H OH
N
33c 1 $ 33c . A solution of 33b in AcOH is treated with H2 (5 psi) over Pd black for 20 h. The reaction mixture is filtered and concentrated in vacuo to yield 33c.
NH H~ NBOC OH
N
N
I
H OH
N
33d 33d. An equimolar solution of 3 3 c and ethyl acetimidate hydrochloride in EtOH is stirred for 18 h.
The reaction solution is concentrated in vacuo to yield a WO 95/24382 218 4 b 91 PCT/US95/02669 white foam. This material is purified by reversed phase HPLC to yield 33d.
33. To a stirred solution of 33d in AcOH (glacial) is added HCl (6.95 M in dioxane). The resulting solution is stirred for 2 h. The solution is concentrated in vacuo and triturated with diethyl ether to yield 33.
Example 34 N-[5S-amino-5-(2,2-dimethyl-1,3-dioxolan-4-yl)pentyl]ethanimidamide, dihydrochloride ~N

H O
Me Me 34. 3B (10 mmol) is dissolved in DMF and 2,2-dimethoxypropane (20 mmol) is added, as is 6N HC1 in dioxane (2 mmol). The mixture is protected from moisture and 2 0 stirred overnight . It is then stripped to a residue in a rotary evaporator using an oil pump as a vacuum source. The residue is suspended in dry acetone, stirred for 30 min, and stripped again. The resulting residue is dissolved in cold water, shelled, and lyophilized to give 34.
Example 35 N-[5S-amino-5-(2-phenyl-1,3-dioxolan-4 yl)pentyl]ethanimidamide, di(4 methylbenzenesulfonate) WO 95124382 218 4 b 91 PCT/US95/02669 ~N
I 2 TsOH O
H O
H
35. 3B ( 27.7 mmol), 1,1,1-trichloroethane (350 mL), 5 benzaldehyde (55.4 mmol), and toluensulfonic acid monohydrate (55.4 mmol) are placed in a 500 mL round bottom single neck flask fitted with a Soxhlet extractor whose thimble is filled with 5A molecular sieves (8-12 mesh beads). The flask is immersed in an oil bath (bath 10 temperature 120 °C) and the mixture is refluxed with vigorous stirring for 16 h. The reaction is then cooled and the mixture is stripped to a residue in a rotary evaporator using an oil pump as a vacuum source. The residue is dissolved in cold water, shelled, and lyophilized to give 1 5 35 .
Example 36 methyl 2-[[3S-amino-2-hydroxy-7-[(1-iminoethyl)amino]heptyl]oxy]propanoate, 2 0 dihydrochloride NH NH2 Me Me"N O OMe I
H 2 HCl OH O

NHBOC Me Z~ home N O
H OH O

WO 95/24382 218 4 6 91 pCT~S95/02669 36a 36a . Sodium hydride (50 o in mineral oil, 10. 5 mmol) is washed twice with hexane and suspended in DMF. Methyl lactate (10 mmol) is dissolved in DMF and added carefully to the NaH suspension with stirring. The mixture is stirred for 30 min, and a solution of 23a (9 mmol) and anhydrous zinc chloride (9 mmol) in THF is added. This mixture is immersed in a 60 °C oil bath and stirred overnight. It is then worked up to give 36a.
NHBOC Me HEN O OMe I
H OH O
36b 1 5 3 6b . A solution of 36a in AcOH is treated with H2 ( 5 psi) over Pd black for 20 h. The reaction mixture is filtered and concentrated in vacuo to yield 36b.
NH NHBOC Me H2N~N O OMe I
H ' OH O
36c 36c. An equimolar solution of 36b and ethyl acetimidate hydrochloride in EtOH is stirred for 18 h.
The reaction solution is concentrated in vacuo to yield a 2 5 white foam. This material is purified by reversed phase HPLC to yield 36c.
36. To a stirred solution of 36c in AcOH (glacial) is added HC1 (6.95 M in dioxane) . The resulting solution is stirred for 2 h. The solution is concentrated in vacuo and triturated with diethyl ether to yield 36.
Example 37 N-[5S-amino-5-(2-methyl-3-oxo-1,4-dioxan-5-yl)pentyl]ethanimidamide, dihydrochloride Me"N O
I
H O
2 HCl Me O

37. A solution of 36 in 1 M aqueous HC1 is refluxed for two hours. The reaction is stripped to small volume, shelled, and lyophilized to give the title compound.
Example 38 N-[5S-amino-6-hydroxy-7-(2-hydroxyphenyl)heptyl]ethanimidamide, dihydrochloride ~N \
I

NHBOC OTHP
Z
~N ~ \
H OH
38a 38a. 2-(Tetrahydropyran-2-yloxy)phenyllithium is prepared from 2-(tetrahydropyran-2-yloxy)phenyl bromide and n-BuLi in THF at -78 °C. It is then reacted with 23a in THF at -78 °C, allowing the temperature to rise to ambient temperature. The reaction mixture is worked up in the usual way to yield 38a.
NHBOC OTHP
H
~N I
I
H OH
38b 38b. A solution of 38a in AcOH is treated with H2 (5 psi) over Pd black for 20 h. The reaction mixture is filtered and concentrated ~n vacuo to yield 38b.
NH NHSOC OTHP

N v \
v ~

I I
OH

38c 38c. An equimolar solution of 38b and ethyl acetimidate hydrochloride in EtOH is stirred for 18 h.
The reaction solution is concentrated 3n vacuo to yield a white foam. This material is purified by reversed phase 2 0 HPLC to yield 38c.
38. To a stirred solution of 38c in AcOH (glacial) is added HCl ( 6. 95 M in dioxane) . The resulting solution is stirred fox 2 h. The solution is concentrated jn vacuo and 2 5 triturated with diethyl ether to yield 3$.
example 39 N-[5S-amino-7,7,7-trifluoro-6-hydroxy-6 methylheptyljethanimidamide WO 95/24382 218 4 6 91 PCT~S95/02669 ~N CF3 OH
H
39a. To a stirring solution of CF3I (10 mmol) in 5 mL
of DMF at -4 0 ° C is added Zn ( 10 mg-atm) and 14a ( 0 . 5 mmol ) in 10 mL of DMF. After stirring for 1 h at -20 °C, the reaction is warmed to room temperature and partitioned between H20 and EtOAc. The organic layer is worked up in the usual manner to obtain desired trifluoromethyl alcohol.
3 9. Using conditions described in example 3, the desired compound is obtained.
Example 40 N-(SS-amino-6-hydroxyheptyl)ethanimidamide, dihydrochloride dihydrate NH OH

40A, 40B
4 Oa . To a 50 mL solution of 14a ( 1 . 5 g, 4 . 0 mmol) in 2 0 EtOH was added NaBHq. After 2h, the reaction was concentrated under vacuum. The residue was taken up in 50 mL of EtOAc and 30 mL of H20. The organic was treated in the usual manner to obtain 1.5 g of 40a.
2 5 40A,40B. Examples 40A, 40B were prepared in same manner as described in example 15. The first eluting fractions from the final purification by reverse phase chromatography were a single isomer (40A). The second eluting fractions were a mixture of two isomers (40B). 40A Analysis calcd.

for C9H21N30 ~ 2 HC1 ~ 2 H20 : C, 3 6 . 4 9; H, 9 . 19; N, 14 . 18 .
Found: C, 36.73; H, 8.93; N, 14.13. 40B Analysis calcd.
for CgH21N30~3 HC1~2 H20: C, 32.49; H, 8.48; N, 12.63.
Found: C, 32.37; H, 8.08; N, 12.04.
S
Example 41 N-[5S-amino-5-(1H-tetrazol-5 yl)pentyl]ethanimidamide, dihydrochloride N/ ~ N
NH
N
H
N v r ~NH2 41a . To a stirring solution of N-oc-Boc-N-E-Z-L-Lys . (3 . 8 g, 10 mmol), 2-aminopropionitrile fumarate (1.9 g, 10 mmol), 1-hydroxybenzotriazole hydrate (4.4 g, 10 mmol), and NMM
1 S (3 .3 mL, 30 mmol) in 50 mL of DMF cooled in an ice bath was added (1H-1,2,3-benzotriazol-1-yloxy)-tris(dimethylamino)phosphonium hexafluorophosphate (BOP) (4.4 g, lOmmol). After stirring 18 h at ambient temperature, the filtrate was concentrated under vacuum.
2 0 The residue was distributed between 100 mL of EtOAc and 50 mL of 1M KHS04 solution. The layers were separated. The organic layer was washed with 1x 50 mL of saturated KHC03 solution and 1x 50 mL of brine and was worked up in the usual manner giving 3.9 g of 41a.
41b . To a stirring solution of 41a (3 . 9 g, 9 mmol) in 90 mL of THF was added PPh3, DEAD, TMSN3. After 24 h of stirring at ambient temperature, the reaction was cooled to 0 'C to which was added slowly 300 mL of 6o Ce(NHq)2(N03)6.
~ 3 0 Additional Ce (NH4) 2 (N03) 6 was added until evolution of N2 ceased. The layers were separated and the aqueous layer was W0.95/24382 218 4 6 91 PCT~S95/02669 extracted 2x 250 mL of DCM. The combined organic layers were treated in the usual manner to yield 3.1 g of 41b.
41c . To a stirring solution of 41b (2 . 7 g, 5 . 9 mmol) in 60 mL of THF was added 7 mL of 1N NaOH. After 18 h, the reaction was concentrated under vacuum. The residue was taken up in 50 mL of EtOAC and 50 mL of 0.5N NaOH. The layers were separated and the aqueous layer was washed 2x 50 mL of EtOAc . The aqueous layer was acidified to pH 3 and extracted 3x 40 mL EtOAc. The second organic extractions were worked up in the usual manner to obtain 0.3 g of 41c.
The original organic extracts were worked up in the usual manner and they also contained product (2 g).
41. To obtain example 41, conditions described in example 3 were used.
Example 42 (A) methyl 3S-amino-2S-hydroxy-7-[(1-iminoethyl)amino]heptanoate .(B) methyl 3S-amino-2R-hydroxy-7-[(1 iminoethyl)amino]heptanoate N H NH2 O , ~ NH2 O
~N O~ N Oi H OH H OH
2 5 42A, 42B
NH Z
Boc NH H
O
42a 3 0 42a . To a stirring solution of N-oc-Z-N-E-Boc-L-Lys-N(OMe)Me lla in ether at 0°C is added LiAlHq (1.2 equiv.) in portions. The resulting solution is stirred for 1h at WO 95/24382 218 4 b 91 PCT/US95/02669 0°C, then carefully quenched with KHS04 (1 M). The layers are separated and the aqueous extracted with ether. The combined organic solutions are extracted with KHSOq (1 M) and NaHC03 (satd.) dried (Na2S04) and evaporated to yield the aldehyde 42a, which is used directly in the next step.
NH Z
CN
Boc NH
OH
42b 42b. A stirred mixture of the aldehyde 42a in EtOAc and KCN (1 equiv.) in water, at 0°C, is treated with an aqueous solution of NaHS03 (satd.). The solution is stirred for 1 h, and the layers separated. The organic solution is dried over Na2SOq anhydrous and concentrated to yield the resulting cyanohydrin 42b.
ZNH O

OH
42c 42c. The cyanohydrin 42b is treated with methanolic/HCl to yield the methyl ester 42c.
42A,42B. The amine 42c is treated with methyl acetimdate 2 5 according to the procedure for 11e. The product is then treated with Pd black according to the procedure for 11, and the isomers separated on reversed phase HPLC to yield 42A and 428.
Example 43 WO 95/24382 ~ ~ ~. 4 b 91 PCT/LTS95/02669 (A) 3S-amino-2S-hydroxy-7-[(1-iminoethyl)amino]heptanoic acid (B) 3S-amino-2R-hydroxy-7-[(I-iminoethyl)amino]heptanoic acid NH NHp O ~ NH2 O
~N OH N OH
H H OH
OH
43A, 43B
ZNH O

OH
43a 43a . The cyanohydrin 42b is treated with concentrated HC1 at 0°C for 12 h. The solution is concentrated under vacuum and the ammonium chloride is removed by filtration.
The residue is then dried to yield the hydroxy acid 43a.
43A,438. The amine 43a is treated with methyl acetimidate 2 0 according to the procedure for 11e. The product is then treated with Pd black according to the procedure for 11, and the isomers separated on reversed phase HPLC to yield 43A and 43B.
2 5 Example 44 methyl 3S-amino-7-[(1-iminoethyl)amino]-2 oxoheptanoate N Oi H O

WO 95/24382 218 4 6 91 pCT~S95/02669 44. A solution of 42A and 42B in water is treated with Mn02. The solution is filtered and concentrated to yield 44.
Example 45 methyl 3-amino-4-[3 [(aminoiminomethyl)amino]phenyl]butanoate H2N N ~
H

O
N02 ~ O~
45a 45a. To a stirred solution of oxalyl chloride (1.1 2 0 equiv.) in dry CH2C12 is slowly added at -60°C a solution of dry DMSO in CH2C12. After the solution is stirred for 5 min a solution of 3-nitrophenethyl alcohol (1 equiv.) in dry CH2C12 is added. The solution is stirred for an additional 15 min and subsequently TEA is added. After stirring for 5 2 5 min the cooling bath is removed and the solution is allowed to reach room temperature. The reaction is quenched with water. The organic layer is removed and the aqueous layer extracted with additional CH2C12. The combined organic extracts are washed with brine and water, dried( Na2S04) and 3 0 evaporated to yield the aldehyde. The aldehyde is treated with carbomethoxymethyl)triphenylphosphonium bromide by the method of 17a, to yield the ester 45a.
NHBoc N02 ~ C02CH3 45b 45b. A solution of 45a and ammonium chloride (3 equiv.) in glacial acetic acid is refluxed for 3 h. The 10 solvent is removed in vacuo and the residue is partitioned between EtOAc and aqueous Na2C03. The layers are separated and the organic phase is dried (Na2SOq) and evaporated to yield the amine. The residue is taken up in THF and treated with di-t-butyl dicarbonate (1.5 equiv.) and triethylamine 15 (1.1 equiv.) . The resulting solution is refluxed for 2 h, concentrated in vacuo and purified by flash column chromatography to yield 45b.
NHBoc NH ~ I C02CH3 20 45c 45c . A solution of 45b in methanol is hydrogenated in the presence of 10~ Pd/C to yield 45c.
2 5 45. Aminoiminomethanesulphonic acid (1.1 equiv.) is added to a solution of 45c in methanol. The solution is stirred for 24 h. The solvent is removed and the residue is dissolved in water. The pH is adjusted to greater than 7 with NaOH. The mixture is extracted with EtOAc, dried 30 (Na2SOq) and concentrated in vacuo. The residue is the treated with methanol/HC1 to yield 45.

W0,95124382 218 4 6 91 PCT/US95/02669 Example 46 3-amino-4-[3 [(aminoiminomethyl)amino]phenyl]butanoic acid ~ I NH2 O
H2N N ~ O H
S H

46. 45 is dissolved in 2N HC1 and refluxed for 1 h. The reaction is diluted with water and lyophilized to yield 46.
Example 47 N-[5S-amino-6-hydroxy-6-(2 hydroxyphenyl)hexyl]ethanimidamide N H NH2 ~
/ 'N
H OH OH

NHBoc Z ~N
O H OTHP
H
47a 47a. 2-(Tetrahydropyran-2-yloxy)phenyllithium is prepared from 2-(tetrahydropyran-2-yloxy)phenyl bromide and n-BuLi in THF at -78 °C. It is then reacted with 3b in THF
2 5 at -78 °C, allowing the temperature to rise to ambient temperature. The reaction mixture is worked up in the usual way to yield 47a.

WO 95/24382 218 4 b 91 PCT/LTS95/02669 NHBoc ~
HEN
OTHP
H OH
47b 47b. A solution of 47a in AcOH is treated with H2 (5 psi) over Pd black for 20 h. The reaction mixture is filtered and concentrated in vacuo to yield 47b.
NH NHBoc ~
~N
OTHP
H OH
47c 47c. An equimolar solution of 47b and ethyl acetimidate hydrochloride in EtOH is stirred for 18 h.
The reaction solution is concentrated in vacuo to yield a white foam. This material is purified by reversed phase HPLC to yield 47c.
47. To a stirred solution of 47c in AcOH (glacial) is added HC1 (6.95 M in dioxane). The resulting solution is 2 0 stirred for 2 h. The solution is concentrated in vacuo and triturated with diethyl ether to yield 47.
Example 48 N-(5S-amino-6-hydroxyhexyl)ethanimidamide NH OH

H

2184b91 48. Example 48 was prepared using methods described in example 3 starting with 3c. h.r.m.s. CgH1gN30: 174.16.
Example 49 N-[5-amino-5-(5-methyloxazol-2-yl)pentyl]ethanimidamide, hydrochloride hydrate / 'N O
I
H NI

NH Boc Z~ N
N
I
H O
49a To a cooled (0°C) solution of N-oc-Boc-N-~-Z-Lys (3.8 g, 10 mmol) , propargylamine (550 mg, 10 mmol) and EtgN (1 g, 10 mmol) in DMF was added HOBT (1.35 g, 10 mmol) and EDC (1.92 g, 10 mmol). The solution was allowed to gradually warm to 2 0 RT over 16 h. EtOAc (500 mL) was added to the reaction solution followed by extraction with brine ( 4 x 100 mL), dried (Na2S04) and concentrated to yield an oil. The product was crystallized from ether/hexane to yield 49a (4.3 g) as a white solid. Anal. Calcd for C22H31N302~ C.
2 5 63.29; H, 7.48; N,10.06. Found: C, 63.04; H, 7.41; N. 9.94.
NH Boc ZwN O
H
I N
49b A solution of 49a (1.8 g, 4.3 mmol) and mercuric acetate (240 mg, .8 mmol) in AcOH (80 mL) was refluxed for 2 h. The solution was stirred at RT for 2h. The solvent was removed S and the residue taken up in CHC13 (250 mL) and washed with NaOH (1M, 1 x 100 mL) and brine (100 mL) , dried (Na2SOq) and evaporated to yield an oil. The product was purified by flash chromatography to yield 49b. Anal. Calcd for C22H31N302 -2 H20: C, 62.75; H, 7.52; N,9.98. Found: C, 62.40; H, 7.40; N. 9.61.
NH Boc HEN O
I
H NI
49c A solution of 49b (2.0 g, 4.75 mmol) in ethanol was treated 1S with H2 (5 psi) over Pd/C (100) for 3 h. The reaction mixture was filtered and concentrated in vacuo to yield 49c.
N H NH Boc / _N O
I
H NI
49d An equimolar solution of 49c (1.0 g, 3.5 mmol)and ethyl acetimidate hydrochloride in EtOH was stirred for 18 h.
2 S The reaction solution is concentrated in vacuo to yield a white foam (1.2 g).
49. To a stirred solution of 49d (1.2 g, 3.3 mmol) in AcOH
(glacial, 25 mL) was added HC1 ( 5.8 M in dioxane, 3 mL).
3 0 The resulting solution was stirred for 1 h. The solution is WO 95/24382 218 4 6 91 pCT~S95/02669 concentrated in vacuo and purified by reversed phase HPLC
to yield 49. Anal. Calcd for C11H20N40 '2.1 HC1 -1.6 H20: C:
40.07; H: 7.74; N:16.99; C1:22.58. Found: C: 40.33; H:
8.00; N: 16.68, C1: 22.75.

Example 50 2S-amino-N-hydroxy-6-[(1 iminoethyl)amino]hexanamide, hydrochloride NOH
N

50a. Starting with N-oc-Z-N-E-Boc-Lys and O-benzylhydroxylamine hydrochloride example 50 was synthesized using conditions described in examples 11a, 15 llc-e, 11. Anal. Calcd for CgHlgNq02~1.5 HC1-0.25 HOAc -H20: C: 35.21; H: 7.82; N:19.32. Found: C: 35.32; H: 7.81;
N: 19.75.
Biological Data The activity of the above listed compounds as NO
synthase inhibitors has been determined in the following assays:
2 5 Citrulline Assay for Nitric Oxide Synthase Nitric oxide synthase activity was measured by monitoring the conversion of L-[2,3-3H]-arginine to L-[2,3-3H]-citrulline (1,2). Human inducible NOS (hiNOS), human endothelial constitutive NOS (hecNOS) and human neuronal constitutive NOS (hncNOS) were each cloned from RNA
extracted from human tissue. The recombinant enzymes were expressed in insect cells using a baculovirus vector.
Enzyme activity was isolated from cell extracts and partially purified by DEAE-SepharoseTM chromatography (2).
Mouse inducible NOS (miNOS) was prepared from an extract of S LPS-treated mouse RAW 264.7 cells and rat brain constitutive NOS (rcNOS) was prepared from an extract of rat cerebellum.
Both preparations were partially purified by DEAF-SepharoseTM
chromatography (2). Enzyme and inhibitors were added to give a volume of 50 ~,L in 50 mM Tris (pH 7.6) and the reaction initiated by the addition of 50 ~.L of a solution containing 50mM Tris (pH 7.6), 2.0 mg/mL bovine serum albumin, 2.0 mM DTT, 4.0 mM CaCl2, 20 ~,M FAD, 100 ~.lM
tetrahydrobiopterin, 0.4- 2.0 mM NADPH and 60 E1M L-arginine containing 0.9 ~.Ci of L-[2,3-3H]-arginine. For 1S constitutive NOS, calmodulin was included at a final concentration of 40-100 nM. Following incubation at 37'C
for 15 minutes, the reaction was terminated by addition of 300 ~,L cold buffer containing 10 mM EGTA, 100 mM HEPES
(pH5.5) and 1.0 mM L-citrulline. The [3H]-citrulline was 2 0 separated by chromatography on DowexTM 50W X-8 cation exchange resin and radioactivity quantified with a liquid scintillation counter.
1. Bredt, D. S. and Snyder, S. H. (1990) Proc. Natl.
2 S Acad . Sci . U. S. A. 87, 682-685 .
2. Misko, T. P., Moore, W. M., Kasten, T. P., Nickols, G.
A., Corbett, J. A., Tilton, R. G., McDaniel, M. L., Williamson, J. R. and Currie, M. G. (1993) Eur. J. Pharm.
233, 119-125.
Raw Cell Ni~~~ sax RAW 264.7 cells are plated to confluency an a 96-well tissue culture plate grown overnight (17h) in the presence 3 S of LPS to induce NOS. A row of 3-6 wells are left untreated WO 95/24382 218 4 6 91 p~~s95/02669 and served as controls for subtraction of nonspecific background. The media is removed from each well and the cells are washed twice with Krebs-Ringers-Hepes (25mM, pH
7.4) with 2 mg/ml glucose. The cells are then placed on ice and incubated with 50 ~,L of buffer containing L-arginine (30 ~.l.M) +/- 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. All values are the average of triplicate wells and are compared to a background-subtracted induced set of cells (1000 value).
The following examples were assayed with the following results.

WO 95/24382 218 4 b 91 PCT/US95/02669 ExamplehiNOS hecNOS hncNOS RAw o inhibiton number IC50 IC50 IC50 cell iNOS at 100 (~1.M) ~N~M) ~IIM) 1-NOS ~,lM

1 d 35 2d 5 3 B 12.3 8420 150 80 15A 9.3 2350 99.6 57.3 16B 76.8 5430 670 >100 3A 53.4 48 40.6 40A 32.6 ' 41 28.5 49 19.4 40B 14.5 11B 4.6 lOf 7.7 ~ 3.4 WO 95/24382 218 4 b 91 PCT~S95/02669 50 40. 6 *At 100 ~,1M dose, response was not seen.
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 5 thereof, can make various changes and modifications of the invention to adapt it to various usages and conditions.

Claims (28)

What is claimed:

1. A compound or a pharmaceutically acceptable salt, prodrug or ester thereof having the formula:
Y is hydrogen, lower alkyl radical, lower alkenyl radical, lower alkynyl radical, aromatic hydrocarbon radical, or alicyclic hydrocarbon radical, wherein all said radicals may optionally be substituted with hydrogen, lower alkyl, nitro, amino, alicyclic hydrocarbon radicals, or aromatic hydrocarbon radicals which may be,optionally substituted with lower alkyl;
X is lower alkylene radical, lower alkenylene radical, lower alkynylene radical, aromatic hydrocarbon radical, (CH2)m Q(CH2)n, where m=1-3, n =1-3, and Q is sulfur, sulfinyl, sulfonyl, oxygen, carbonyl, lower alkynyl radical, aromatic hydrocarbon radical, or alicyclic hydrocarbon radical wherein all said radicals are optionally substituted with hydrogen, halogen and lower alkyl;
R1, R2, R3 and R4 are independently selected from the group consisting of hydrogen and lower alkyl;
A is lower alkylene radical, lower alkenylene radical, lower alkynylene radical, alicyclic hydrocarbon radical, carbonyl, or aromatic hydrocarbon radical, wherein all said radicals are optionally substituted with hydrogen, lower alkyl, hydroxyl, lower alkoxy, alkoxycarbonyl, alkylaryloxy, thiol, lower thioalkoxy, thioalkylaryloxy, thioaryloxy, sulfinylalkyl, sulfinylalkylaryl, sulfinylaryl, sulfonylalkyl, sulfonylalkylaryl, sulfonylaryl, halogen, aromatic hydrocarbon radicals, or alicyclic hydrocarbon radicals;
B is hydrogen, lower alkyl radical, lower alkenyl radical, lower alkynyl radical, lower alkoxy radical, hydroxy, alkylaryloxy, thiol, lower thioalkoxy, lower thioalkylaryloxy, thioaryloxy, sulfinylalkyl, sulfinylalkylaryl, sulfinylaryl, sulfonylalkyl, sulfonylalkylaryl, sulfonylaryl, aromatic hydrocarbon radical, alicyclic hydrocarbon radical, or NHOH, wherein all said radicals are optionally substituted with hydrogen, lower alkyl, hydroxyl, lower alkoxy, halogen, aromatic hydrocarbon radicals, or alicyclic hydrocarbon radical;
with the proviso that when A is CHOR, B cannot.be OR and Y cannot be amino wherein R is alkyl;
with the proviso that when A is carbonyl, B is not hydroxy, aryloxy, thioalkoxy, alkoxy; or NHOH.

2. The compound as recited in claim 1 wherein;
Y is hydrogen, lower alkyl radical, lower alkenyl radical, lower alkynyl radical, aromatic hydrocarbon radical, or alicyclic hydrocarbon radical, wherein all said radicals may optionally be substituted with hydrogen, cyano, lower alkyl, nitro, amino, alicyclic hydrocarbon radicals, or aromatic hydrocarbon radicals which may be optionally substituted with lower alkyl;
X is lower alkylene radical, lower alkenylene radical, lower alkynylene radical, aromatic hydrocarbon radical, (CH2)m Q(CH2)n, where m=1-3 , n =1-3, and Q is sulfur, sulfinyl, sulfonyl, oxygen, carbonyl, lower alkynyl radical, aromatic hydrocarbon radical, or alicyclic hydrocarbon radical, wherein all said radicals are optionally substituted with hydrogen, halogen and lower alkyl;
R1, R2, R3 and R4 are independently selected from the group consisting of hydrogen and lower alkyl;
A is lower alkylene radical, lower alkenylene radical, lower alkynylene radical, alicyclic hydrocarbon radical, or aromatic hydrocarbon radical, wherein all said radicals are optionally substituted with hydrogen, lower alkyl, hydroxyl, lower alkoxy, alkylaryloxy, thiol, lower thioalkoxy, thioalkylaryloxy, thioaryloxy, sulfinylalkyl, sulfinylalkylaryl, sulfinylaryl, sulfonylalkyl, sulfonylalkylaryl, sulfonylaryl, halogen, aromatic hydrocarbon radicals; or alicyclic hydrocarbon radicals;
B is hydrogen, lower alkoxy radical, hydroxy, alkylaryloxy, thiol, lower thioalkoxy, lower thioalkylaryloxy, thioaryloxy, sulfinylalkyl, sulfinylalkylaryl, sulfinylaryl, sulfonylalkyl, sulfonylalkylaryl, sulfonylaryl, or NHOH.

3. A compound or a pharmaceutically acceptable salt, prodrug or ester thereof having the formula:
wherein; Y is hydrogen or lower alkyl which may optionally be substituted with hydrogen, lower alkyl, nitro, amino, alicyclic hydrocarbon radicals, or aromatic hydrocarbon radicals which may be optionally substituted with lower alkyl;
X is lower alkylene from 3-5 carbons which may optionally be substituted with hydrogen, halogen and lower alkyl;
R1, R2, R3 and R4 are independently selected from the group consisting of hydrogen and lower alkyl;
A is lower alkylene from 2-4 carbons optionally substituted with hydroxyl; and B is hydroxyl.

4. The compound as recited in claim 3 wherein;
Y is methyl;
X is butylene;
R1, R2, R3 and R4 are hydrogen;

A is ethylene or isopropylene optionally substituted with hydroxyl; and B is hydroxyl.

5. The compound as recited in claim 1 wherein;
A is lower alkyl substituted with hydrogen, lower alkyl, hydroxyl, lower alkoxy, halogen, aromatic hydrocarbon radicals, or alicyclic hydrocarbon radicals.

6. The compound as recited in claim 1 wherein the compound is selected from the group consisting of;
N1-(1-iminoethyl)-1,4-pentanediamine;
N1-(1-iminoethyl)-1,5-heptanediamine;
N1-(1-iminoethyl)-5-phenyl-1,5-pentanediamine;
N-[5-amino-5-(2-hydroxyphenyl)pentyl]ethanimidamide;
N-[5-amino-5-(4-hydroxyphenyl)pentyl]ethanimidamide;
N-(5-aminononyl)ethanimidamide;
N-(5S-amino-6-oxoheptyl)ethanimidamide, dihydrochloride;
N-(5S-amino-6,7-dihydroxy-6-methylheptyl)ethanimidamide, hydrochloride dehydrate;
N-(5S-amino-6,7-dihydroxyoctyl)ethanimidamide, dihydrochloride hydrate;
N-(5S-amino-6,7,8-trihydroxyoctyl)ethanimidamide;
N-(5S-amino-7,8-dihydroxyoctyl)ethanimidamide;
N-(5S-amino-6-hydroxy-7-methoxyheptyl)ethanimidamide;
N-[5S-amino-6-hydroxy-7-(ethylthio)heptyl]ethanimidamide;
N-[5S-amino-6-hydroxy-7-methylsulfinyl)heptyl]
ethanimidamide;
N-[5S-amino-6-hydroxy-7-methylsulfonyl)heptyl]
ethanimidamide;
N-[5S-amino-6-hydroxy-7-(phenylmethyl)thio]heptyl]
ethanimidamide;
N-[5S-amino-6-hydroxy-7-[(phenylmethyl) sulfinyl]heptyl]ethanimidamide;
N-[5S-amino-6-hydroxy-7-[(phenylmethyl)sulfonyl]
heptyl]ethanimidamide;
N-(5S-amino-6-fluoro-7-hydroxy-6-methylheptyl) ethanimidamide;
N-[5S-amino-6-hydroxy-7-(2-hydroxyphenyl)heptyl]
ethanimidamide, dihydrochloride;
N-[5S-amino-7,7,7-trifluoro-6-hydroxy-6-methylheptyl) ethanimidamide;

N-(5S-amino-6-hydroxyheptyl)ethanimidamide, dihydrochloride dehydrate;
N-[5S-amino-6-hydroxy-6-(2-hydroxyphenyl)hexyl]
ethanimidamide;
N-(5S-amino-6-hydroxyhexyl)ethanimidamide; and .alpha. -[1-amino-5-[(1-iminoethyl)amino]pentyl]benzenemethanol hydrochloride dehydrate.

7. The compound as recited in claim 3 wherein the compound is selected from the group consisting of N-(5S-amino-6,7-dihydroxyheptyl)ethanimidamide, dihydrochloride; and N-(5S-amino-6,7-dihydroxy-6-methylheptyl)ethanimidamide, hydrochloride dehydrate.

8. A pharmaceutical composition comprising a compound as recited in claim 1 together with a pharmaceutically acceptable carrier.

9. A pharmaceutical composition comprising a compound as recited in claim 2 together with a pharmaceutically acceptable carrier.

10. A pharmaceutical composition comprising a compound as recited in claim 3, 4, 5, 6, or 7 together with a pharmaceutically acceptable carrier.

11. Use of a therapeutically effective amount of a compound having the formula:

wherein: Y is hydrogen, lower alkyl radical, lower alkenyl radical, lower alkynyl radical, aromatic hydrocarbon radical, or alicyclic hydrocarbon radical, wherein all said radicals may optionally be substituted with hydrogen, lower alkyl, nitro, amino, alicyclic hydrocarbon radicals, or aromatic hydrocarbon radicals which may be. optionally substituted with lower alkyl;
X is lower alkylene radical, lower alkenylene radical;
lower alkynylene radical, aromatic hydrocarbon radical, (CH2)m Q(CH2)n, where m=1-3 , n=1-3 , and Q is sulfur, sulfinyl, sulfonyl, oxygen, carbonyl, lower alkynyl radical, aromatic hydrocarbon radical, or alicyclic hydrocarbon radical wherein all said radicals are optionally substituted with hydrogen, halogen and lower alkyl;
R1, R2, R3 and R4 are independently selected from the group consisting of hydrogen and lower alkyl;
A is lower alkylene radical, lower alkenyle radical, lower alkynylene radical, alicyclic hydrocarbon radical, carbonyl, or aromatic hydrocarbon radical, wherein all said radicals are optionally substituted with hydrogen, lower alkyl, hydroxyl, lower alkoxy, alkoxycarbonyl, alkylaryloxy, thiol, lower thioalkoxy, thioalkylaryloxy, thioaryloxy, sulfinylalkyl, sulfinylalkylaryl, sulfinylaryl, sulfonylalkyl, sulfonylalkylaryl, sulfonylaryl, halogen, aromatic hydrocarbon radicals, or alicyclic hydrocarbon radicals;
B is hydrogen, lower alkyl radical, lower alkenyl radical, lower alkynyl radical, lower alkoxy radical, hydroxy, alkylaryloxy, thiol, lower thioalkoxy, lower thioalkylaryloxy, thioaryloxy, sulfinylalkyl, sulfinylalkylaryl, sulfinylaryl, sulfonylalkyl, sulfonylalkylaryl, sulfonylaryl, aromatic hydrocarbon radical, alicyclic hydrocarbon radical, or NHOH, wherein all said radicals are optionally substituted with hydrogen, lower alkyl, hydroxyl, lower alkoxy, halogen, aromatic hydrocarbon radicals, or alicyclic hydrocarbon radical;
with the proviso that when A is CHOR, B cannot be OR and Y cannot be amino wherein R is alkyl;
with the proviso that when A is carbonyl, B is not hydroxy, aryloxy, thioalkoxy, alkoxy, or NHOH;
for inhibiting nitric oxide synthesis in a subject in need of such inhibition.

12. The use according to claim 11 wherein:
Y is hydrogen, lower alkyl radical, lower alkenyl radical, lower alkynyl radical, aromatic hydrocarbon radical, or alicyclic hydrocarbon radical, wherein all said radicals may optionally be substituted with hydrogen, cyano, lower alkyl, nitro, amino, alicyclic hydrocarbon radicals, or aromatic hydrocarbon radicals which may be optionally substituted with lower alkyl;
X is lower alkylene radical, lower alkenylene radical, lower alkynylene radical, aromatic hydrocarbon radical, (CH2)m Q(CH2)n, where m=1-3, n=1-3, and Q is sulfur, sulfinyl, sulfonyl, oxygen, carbonyl, lower alkynyl radical, aromatic hydrocarbon radical, or alicyclic hydrocarbon radical, wherein all said radicals are optionally substituted with hydrogen, halogen and lower alkyl;
R1, R2, R3 and R4 are independently selected from the group consisting of hydrogen and lower alkyl;
A is lower alkylene radical, lower alkenylene radical, lower alkynylene radical, alicyclic hydrocarbon radical, or aromatic hydrocarbon radical, wherein all said radicals are optionally substituted with hydrogen, lower alkyl, hydroxyl, lower alkoxy, alkylaryloxy, thiol, lower thioalkoxy, thioalkylaryloxy, thioaryloxy, sulfinylalkyl, sulfinylalkylaryl, sulfinylaryl, sulfonylalkyl, sulfonylalkylaryl, sulfonylaryl, halogen, aromatic hydrocarbon radicals, or alicyclic hydrocarbon radicals;
B is hydrogen, -lower alkoxy radical, hydroxy, alkylaryloxy, thiol, lower thioalkoxy, lower thioalkylaryloxy, thioaryloxy, sulfinylalkyl, sulfinylalkylaryl, sulfinylaryl, sulfonylalkyl, sulfonylalkylaryl, sulfonylaryl, or NHOH.

13. Use of a therapeutically effective amount of a compound having the formula:

wherein: Y is hydrogen or lower alkyl which may optionally be substituted with hydrogen, lower alkyl, nitro, amino, alicyclic hydrocarbon radicals, or aromatic hydrocarbon radicals which may be optionally substituted with lower alkyl;
X is lower alkylene from 3-5 carbons which may optionally substituted with hydrogen, halogen and lower alkyl;
R1, R2, R3 and R4 are independently selected from the group consisting of hydrogen and lower alkyl;
A is lower alkylene from 2-4 carbons optionally substituted with hydroxyl; and B is hydroxyl;
for inhibiting nitric oxide synthesis in a subject in need of such inhibition.

14. The use according to claim 13 wherein:
Y is methyl;
X is butylene;

R1, R2, R3 and R4 are hydrogen;
A is ethylene or isopropylene optionally substituted with hydroxyl; and B is hydroxyl.

15. The use according to claim 11 wherein:
A is lower alkylene substituted with hydrogen, lower alkyl, hydroxyl, lower alkoxy, halogen, aromatic hydrocarbon radicals, or alicyclic hydrocarbon radicals.

16. The use according to claim 11 wherein the compound is selected from the group consisting of:
N1-(1-iminoethyl)-1,4-pentanediamine;
N1-(1-iminoethyl)-1,5-heptanediamine;
N1-(1-iminoethyl)-5-phenyl-1,5-pentanediamine;
N-[5-amino-5-(2-hydroxyphenyl)pentyl]ethanimidamide;
N-[5-amino-5-(4-hydroxyphenyl)pentyl]ethanimidamide;
N-(5-aminononyl)ethanimidamide;
N-(5S-amino-6-oxoheptyl)ethanimidamide, dihydrochloride;
N-(5S-amino-6,7-dihydroxy-6-methylheptyl)ethanimidamide, hydrochloride dihydrate;

N-(5S-amino-6,7-dihydroxyoctyl)ethanimidamide, dihydrochloride hydrate;
N-(5S-amino-6,7,8-trihydroxyoctyl)ethanimidamide;
N-(5S-amino-7,8-dihydroxyoctyl)ethanimidamide;
N-(5S-amino-6-hydroxy-7-methoxyheptyl)ethanimidamide;
N-[5S-amino-6-hydroxy-7-(ethylthio)heptyl]ethanimidamide;
N-[5S-amino-6-hydroxy-7-methylsulfinyl)heptyl]
ethanimidamide;
N-[5S-amino-6-hydroxy-7-methylsulfonyl)heptyl]
ethanimidamide;
N-[5S-amino-6-hydroxy-7-(phenylmethyl)thio]heptyl]
ethanimidamide;
N-[5S-amino-6-hydroxy-7-[(phenylmethyl) sulfinyl]heptyl]ethanimidamide;
N-[5S-amino-6-hydroxy-7-[(phenylmethyl)sulfonyl]
heptyl]ethanimidamide;
N-(5S-amino-6-fluoro-7-hydroxy-6-methylheptyl) ethanimidamide;
N-[5S-amino-6-hydroxy-7-(2-hydroxyphenyl)heptyl]
ethanimidamide, dihydrochloride;
N-[5S-amino-7,7,7-trifluoro-6-hydroxy-6-methylheptyl) ethanimidamide;
N-(5S-amino-6-hydroxyheptyl)ethanimidamide, dihydrochloride dehydrate;
N-[5S-amino-6-hydroxy-6-(2-hydroxyphenyl)hexyl]
ethanimidamide;
N-(5S-amino-6-hydroxyhexyl)ethanimidamide; and .alpha. -[1-amino-5-[(1-iminoethyl)amino]pentyl]benzenemethanol hydrochloride dehydrate.

17. Use of a therapeutically effective amount of a compound having the formula:
wherein: Y is hydrogen, lower alkyl radical, lower alkenyl radical, lower alkynyl radical, aromatic hydrocarbon radical, or alicyclic hydrocarbon radical, wherein all said radicals may optionally be substituted with hydrogen, lower alkyl, nitro, amino, alicyclic hydrocarbon radicals, or aromatic hydrocarbon radicals which may be optionally substituted with lower alkyl;
X is lower alkylene radical, lower alkenylene radical, lower alkynylene radical, aromatic hydrocarbon radical, (CH2)m Q(CH2)n, where m=1-3, n=1-3, and Q is sulfur, sulfinyl, sulfonyl, oxygen, carbonyl, lower alkynyl radical, aromatic hydrocarbon radical, or alicyclic hydrocarbon radical wherein all said radicals are optionally substituted with hydrogen, halogen and lower alkyl;
R1, R2, R3 and R4 are independently selected from the group consisting of hydrogen and lower alkyl;
A is lower alkylene radical, lower alkenylene radical, lower alkynylene radical, alicyclic hydrocarbon radical, carbonyl, or aromatic hydrocarbon radical, wherein all said radicals are optionally substituted with hydrogen, lower alkyl, hydroxyl, lower alkoxy, alkoxycarbonyl, alkylaryloxy, thiol, lower thioalkoxy, thioalkylaryloxy, thioaryloxy, sulfinylalkyl, sulfinylalkylaryl, sulfinylaryl, sulfonylalkyl, sulfonylalkylaryl, sulfonylaryl, halogen, aromatic hydrocarbon radicals, or alicyclic hydrocarbon radicals;
B is hydrogen, lower alkyl radical, lower alkenyl radical, lower alkynyl radical, lower alkoxy radical, hydroxy, alkylaryloxy, thiol, lower thioalkoxy, lower thioalkylaryloxy, thioaryloxy, sulfinylalkyl, sulfinylalkylaryl, sulfinylaryl, sulfonylalkyl, sulfonylalkylaryl, sulfonylaryl, aromatic hydrocarbon radical, alicyclic hydrocarbon radical, or NHOH, wherein all said radicals are optionally substituted with hydrogen, lower alkyl, hydroxyl, lower alkoxy, halogen, aromatic hydrocarbon radicals,or alicyclic hydrocarbon radical;
with the proviso that when A is CHOR, B cannot be OR and Y cannot be amino wherein R is alkyl;
with the proviso that when A is carbonyl, B is not hydroxy, aryloxy, thioalkoxy, alkoxy, or NHOH;
for selectively inhibiting nitric 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.

18. The use according to claim 17 wherein:
Y is hydrogen, lower alkyl radical, lower alkenyl radical, lower alkynyl radical, aromatic hydrocarbon radical, or alicyclic hydrocarbon radical, wherein all said radicals may optionally be substituted with hydrogen, cyano, lower alkyl, nitro, amino, alicyclic hydrocarbon radicals, or aromatic hydrocarbon radicals which may be optionally substituted with lower alkyl;
X is lower alkylene radical, lower alkenylene radical, lower alkynylene radical, aromatic hydrocarbon radical, (CH2)m Q(CH2)n, where m=1-3, n=1-3, and Q is sulfur, sulfinyl, sulfonyl, oxygen, carbonyl, lower alkynyl radical, aromatic hydrocarbon radical, or alicyclic hydrocarbon radical, wherein all said radicals are optionally substituted with hydrogen, halogen and lower alkyl;
R1, R2, R3 and R4 are independently selected from the group consisting of hydrogen and lower alkyl;
A is lower alkylene radical, lower alkenylene radical, lower alkynylene radical, alicyclic hydrocarbon radical; or aromatic hydrocarbon radical, wherein all said radicals are optionally substituted with hydrogen, lower alkyl, hydroxyl, lower alkoxy, alkylaryloxy, thiol, lower thioalkoxy, thioalkylaryloxy, thioaryloxy, sulfinylalkyl, sulfinylalkylaryl, sulfinylaryl, sulfonylalkyl, sulfonylalkylaryl, sulfonylaryl, halogen, aromatic hydrocarbon radicals, or alicyclic hydrocarbon radicals;

B is hydrogen, lower alkoxy radical, hydroxy, alkylaryloxy, thiol, lower thioalkoxy, lower thioalkylaryloxy, thioaryloxy, sulfinylalkyl, sulfinylalkylaryl, sulfinylaryl, sulfonylalkyl, sulfonylalkylaryl, sulfonylaryl, or NHOH.

19. Use of a therapeutically effective amount of a compound having the formula:

wherein: Y is hydrogen or lower alkyl which may optionally be substituted with hydrogen, lower alkyl, nitro; amino, alicyclic hydrocarbon radicals, or aromatic hydrocarbon radicals which may be optionally substituted with lower alkyl;

X is lower alkylene from 3-5 carbons which may optionally substituted with hydrogen, halogen and lower alkyl;

R1, R2, R3 and R4 are independently selected from the group consisting of hydrogen and lower alkyl;

A is lower alkylene from 2-4 carbons optionally substituted with hydroxyl; and B is hydroxyl;

for selectively inhibiting nitric 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.

20. The use according to claim 19 wherein:

X is methyl;

X is butylene;

R1, R2, R3 and R4 are hydrogen;

A is ethylene or isopropylene optionally substituted with hydroxyl; and B is hydroxyl.

21. The use according to claim 17 wherein:

A is lower alkylene substituted with hydrogen, lower alkyl, hydroxyl, lower alkoxy, halogen, aromatic hydrocarbon radicals, or alicyclic hydrocarbon radicals.

22. The use according to claim 17 wherein the compound is selected from the group consisting of:
N1-(1-iminoethyl)-1,4-pentanediamine;
N1-(1-iminoethyl)-1,5-heptanediamine;
N1-(1-iminoethyl)-5-phenyl-1,5-pentanediamine;
N-[5-amino-5-(2-hydroxyphenyl)pentyl]ethanimidamide;
N-[5-amino-5-(4-hydroxyphenyl)pentyl]ethanimidamide;
N-(5-aminononyl)ethanimidamide;
N-(5S-amino-6-oxoheptyl)ethanimidamide, dihydrochloride;
N-(5S-amino-6,7-dihydroxy-6-methylheptyl)ethanimidamide, hydrochloride dehydrate;
N-(5S-amino-6,7-dihydroxyoctyl)ethanimidamide, dihydrochloride hydrate;
N-(5S -amino-6,7,8-trihydroxyoctyl)ethanimidamide;
N-(5S-amino-7,8-dihydroxyoctyl)ethanimidamide;
N-(5S-amino-6-hydroxy-7-methoxyheptyl)ethanimidamide;
N-[5S-amino-6-hydroxy-7-(ethylthio)heptyl]ethanimidamide;
N-[5S-amino-6-hydroxy-7-methylsulfinyl)heptyl]
ethanimidamide;

N-[5S-amino-6-hydroxy-7-methylsulfonyl)heptyl]
ethanimidamide;
N-[5S-amino-6-hydroxy-7-(phenylmethyl)thio]heptyl]
ethanimidamide;
N-[5S-amino-6-hydroxy-7-[(phenylmethyl) sulfinyl]heptyl]ethanimidamide;
N-[5S-amino-6-hydroxy-7-[(phenylmethyl)sulfonyl]
heptyl]ethanimidamide;
N-(5S-amino-6-fluoro-7-hydroxy-6-methylheptyl) ethanimidamide;
N-[5S-amino-6-hydroxy-7-(2-hydroxyphenyl)heptyl]
ethanimidamide, dihydrochloride;
N-[5S-amino-7,7,7-trifluoro-6-hydroxy-6-methylheptyl) ethanimidamide;
N-(5S-amino-6-hydroxyheptyl)ethanimidamide, dihydrochloride dihydrate;
N-[5S-amino-6-hydroxy-6-(2-hydroxyphenyl)hexyl]
ethanimidamide;
N-(5S-amino-6-hydroxyhexyl)ethanimidamide; and .alpha. - [1-amino-5-[(1-iminoethyl)amino]pentyl]benzenemethanol hydrochloride dehydrate.

23. Use of a therapeutically effective amount of a compound having the formula:

wherein: Y is hydrogen, lower alkyl radical, lower alkenyl radical, lower alkynyl radical, aromatic hydrocarbon radical, or alicyclic hydrocarbon radical, wherein all said radicals may optionally be substituted with hydrogen, lower alkyl, nitro, amino, alicyclic hydrocarbon radicals, or aromatic hydrocarbon radicals which may be optionally substituted with lower alkyl;

X is lower alkylene radical, lower alkenylene radical, lower alkynylene radical, aromatic hydrocarbon radical, (CH2)m Q(CH2)n, where m=1-3, n=1-3, and Q is sulfur;
sulfinyl, sulfonyl, oxygen, carbonyl, lower alkynyl radical, aromatic hydrocarbon radical, or alicyclic hydrocarbon radical wherein all said radicals are optionally substituted with hydrogen, halogen and lower alkyl;

R1, R2, R3 and R4 are independently selected from the group consisting of hydrogen and lower alkyl;

A is lower alkylene radical, lower alkenylene radical, lower alkynylene radical, alicyclic hydrocarbon radical, carbonyl, or aromatic hydrocarbon radical, wherein all said radicals are optionally substituted with hydrogen, lower alkyl, hydroxyl, lower alkoxy, alkoxycarbonyl, alkylaryloxy, thiol, lower thioalkoxy, thioalkylaryloxy, thioaryloxy, sulfinylalkyl, sulfinylalkylaryl, sulfinylaryl, sulfonylalkyl, sulfonylalkylaryl, sulfonylaryl, halogen, aromatic hydrocarbon radicals, or alicyclic hydrocarbon radicals;

B is hydrogen, lower alkyl radical, lower alkenyl radical, lower alkynyl radical, lower alkoxy radical, hydroxy, alkylaryloxy, thiol, lower thioalkoxy, lower thioalkylaryloxy, thioaryloxy, sulfinylalkyl, sulfinylalkylaryl, sulfinylaryl, sulfonylalkyl, sulfonylalkylaryl, sulfonylaryl, aromatic hydrocarbon radical, alicyclic hydrocarbon radical, or NHOH, wherein all said radicals are optionally substituted with hydrogen, lower alkyl, hydroxyl, lower alkoxy, halogen, aromatic hydrocarbon radicals, or alicyclic hydrocarbon radical;
with the proviso that when A is CHOR, B cannot be OR and Y cannot be amino wherein R is alkyl;
with the proviso that when A is carbonyl, B is not hydroxy, aryloxy, thioalkoxy, alkoxy, or NHOH;
for inhibiting nitric oxide synthesis in a subject in need of such inhibition.

24. The use according to claim 23 wherein:
Y is hydrogen, lower alkyl radical, lower alkenyl radical, lower alkynyl radical, aromatic hydrocarbon radical, or alicyclic hydrocarbon radical, wherein all said radicals may optionally be substituted with hydrogen, cyano, lower alkyl, nitro, amino, alicyclic hydrocarbon radicals, or aromatic hydrocarbon radicals which may be optionally substituted with lower alkyl;
X is lower alkylene radical, lower alkenylene radical, lower alkynylene radical, aromatic hydrocarbon radical, (CH2)m Q(CH2)n, where m=1-3, n=1-3, and Q is sulfur, sulfinyl, sulfonyl, oxygen, carbonyl, lower alkynyl radical, aromatic hydrocarbon radical, or alicyclic hydrocarbon radical, wherein all said radicals are optionally substituted with hydrogen, halogen and lower alkyl;
R1, R2, R3 and R4 are independently selected from the group consisting of hydrogen. and lower alkyl;
A is lower alkylene radical, lower alkenylene radical, lower alkynylene radical, alicyclic hydrocarbon radical, or aromatic hydrocarbon radical, wherein all said radicals are optionally substituted with hydrogen, lower alkyl, hydroxyl, lower alkoxy, alkylaryloxy, thiol, lower thioalkoxy, thioalkylaryloxy, thioaryloxy, sulfinylalkyl, sulfinylalkylaryl, sulfinylaryl, sulfonylalkyl, sulfonylalkylaryl, sulfonylaryl, halogen, aromatic hydrocarbon radicals, or alicyclic hydrocarbon radicals;
B is hydrogen, lower alkoxy radical, hydroxy, alkylaryloxy, thiol, lower thioalkoxy, lower thioalkylaryloxy, thioaryloxy, sulfinylalkyl, sulfinylalkylaryl, sulfinylaryl, sulfonylalkyl, sulfonylalkylaryl, sulfonylaryl, or NHOH.

25. Use of a therapeutically effective amount of a compound having the formula:

wherein: Y is hydrogen or lower alkyl which may optionally be substituted with hydrogen, lower alkyl, nitro, amino, alicyclic hydrocarbon radicals, or aromatic hydrocarbon radicals which may be optionally substituted with lower alkyl;
X is lower alkylene from 3-5 carbons which may optionally substituted with hydrogen, halogen and lower alkyl;
R1, R2, R3 and R4 are independently selected from the group consisting of hydrogen and lower alkyl;
A is lower alkylene from 2-4 carbons optionally substituted with hydroxyl; and B is hydroxyl;
for inhibiting nitric oxide synthesis in a subject in need of such inhibition.

26. The use according to claim 23 wherein:
Y is methyl;

X is butylene;
R1, R2, R3 and R4 are hydrogen;
A is ethylene or isopropylene optionally substituted with hydroxyl and B is hydroxyl.

27. The use according to claim 23 wherein:
A is lower alkylene substituted with hydrogen, lower alkyl, hydroxyl, lower alkoxy, halogen, aromatic hydrocarbon radicals, or alicyclic hydrocarbon radicals.

28. The use according to claim 23 wherein the compound is selected from the group consisting of:
N1-(1-iminoethyl)-1,4-pentanediamine;
N1-(1-iminoethyl)-1,5-heptanediamine;
N1-(1-iminoethyl)-5-phenyl-1,5-pentanediamine;
N-[5-amino-5-(2-hydroxyphenyl)pentyl]ethanimidamide;
N-[5-amino-5-(4-hydroxyphenyl)pentyl]ethanimidamide;
N-(5-aminononyl)ethanimidamide;
N-(5S-amino-6-oxoheptyl)ethanimidamide, dihydrochloride;
N-(5S-amino-6,7-dihydroxy-6-methylheptyl)ethanimidamide, hydrochloride dehydrate;
N-(5S-amino-6,7-dihydroxyoctyl)ethanimidamide, dihydrochloride hydrate;
N-(5S-amino-6,7,8-trihydroxyoctyl)ethanimidamide;
N-(5S-amino-7,8-dihydroxyoctyl)ethanimidamide;
N-(5S-amino-6-hydroxy-7-methoxyheptyl)ethanimidamide;
N-[5S-amino-6-hydroxy-7-(ethylthio)heptyl]ethanimidamide;
N-(5S-amino-6-hydroxy-7-methylsulfinyl)heptyl]
ethanimidamide;
N-(5S-amino-6-hydroxy-7-methylsulfonyl)heptyl]
ethanimidamide;
N-[5S-amino-6-hydroxy-7-(phenylmethyl)thio]heptyl]
ethanimidamide;
N-[5S-amino-6-hydroxy-7-[(phenylmethyl) sulfinyl]heptyl]ethanimidamide;
N-[5S-amino-6-hydroxy-7-[(phenylmethyl)sulfonyl]
heptyl]ethanimidamide;
N-(5S-amino-6-fluoro-7-hydroxy-6-methylheptyl) ethanimidamide;
N-[5S-amino-6-hydroxy-7-(2-hydroxyphenyl)heptyl]
ethanimidamide, dihydrochloride;

N-[5S-amino-7,7,7-trifluoro-6-hydroxy-6-methylheptyl]
ethanimidamide;
N-(5S-amino-6-hydroxyheptyl)ethanimidamide, dihydrochloride dehydrate;
N-[5S-amino-6-hydroxy-6-(2-hydroxyphenyl)hexyl]
ethanimidamide;
N-(5S-amino-6-hydroxyhexyl)ethanimidamide; and .alpha.- [1-amino-5- [ (1-iminoethyl) amino] pentyl] benzenemethanol hydrochloride dehydrate.