CA2184385A1 - Composition for the treatment of lung disease - Google Patents

Abstract

Disclosed are pharmaceutical compositions for the treatment of lung disease, and in particular Cystic Fibrosis, comprising a substituted azetidinone of general formula (I), which have been found to be potent elastase inhibitors and a (F)-actin shortening protein, such as gelsolin.

Description

218438~

- TITLE OF THE INVhNTION
COMPOSITION FOR THE TREATMENT OF LUNG DISEASE

BACKGROUND OF THE INVENTION
This invention is directed to ph~rm~ceutical compositions comprising an elastase inhibitor and an (F)-actin shortening protein such as gelsolin or vitamin D binding protein (DBP), for use in the treatment of Cystic Fibrosis and related diseases. In particular, this invention is directed to ph~rm~ceutical compositions comprising elastase inhibitors Of Formula I
Ra R1 o~/~

O CONHCH ~--R2 and (F)-actin shortening proteins, Gelsolin being preferred.
We have found that a group of new substituted azetidinones are potent elastase inhibitors and therefore are useful anti-infl~mm~tory and antidegenerative agents.
Proteases from granulocytes and macrophages have been reported to be responsible for the chronic tissue destruction mech~ni~m~
25 associated with infl~mm~tion, including rheumatoid al~llilis and emphysema. Accordingly, specific and selective inhibitors of these proteases are candidates for potent anti-infl~mm~tory agents useful in the treatment of infl~mm~tory conditions resulting in connective tissue destruction, e.g., rheumatoid arthritis, emphysema, bronchial 30 infl~mm~tion, chronic bronchitis, glomerulonephritis, osteoallhlilis, spondylitis, lupus, psoriasis, atherosclerosis, sepsis, septicemia, shock, myocardial infarction, re~elrusion injury, periodontitis, cystic fibrosis and acute respiratory distress syndrome.

W O 95/24207 PC~r~US95/02938 21843~5 The role of proteases from granulocytes, leukocytes or macrophages are related to a rapid series of events which occurs during the progression of an infl~mm~tory condition:-(1) There is a rapid production of prostaglandins (PG) and related compounds synthesized from arachidonic acid. This PG
synthesis has been shown to be inhibited by aspirin-related nonsteroidal anti-i~fl~mm~tory agents including indomethacin and phenylbutazone.
There is some evidence that protease inhibitors prevent PG production;

(2) There is also a change in vascular permeability which causes a leakage of fluid into the infl~med site and the resulting edema is generally used as a marker for measuring the degree of infl~mm~tion.
This process has been found to be induced by the proteolytic or peptide cleaving activity of proteases, especially those contained in the granulocyte, and thereby can be inhibited by various synthetic protease inhibitors, for example, N-acyl benzisothiazolones and the respective 1,1-dioxides. Morris Zimmerman et al., J. Biol. Chem., 255, 9848 (1980); and (3) There is an appearance and/or presence of lymphoid cells, especially macrophages and polymorphonuclear leukocytes (PMN). It has been known that a variety of proteases are released from the macrophages and PMN, further indicating that the proteases do play an important role in infl~mm~tion.
In general, proteases are an important family of enzymes within the peptide bond cleaving enzymes whose members are essential to a variety of normal biological activities, such as digestion, formation and dissolution of blood clots, the formation of active forms of hormones, the immune reaction to foreign cells and org~ni~m~, etc., and in pathological conditions such as the degradation of structural proteins 30 at the articular cartilage/pannus junction in rheumatoid arthritis etc.
Elastase is one of the proteases. It is an enzyme capable of hydrolyzing the connective tissue component elastin, a property not contained by the bulk of the proteases present in m~mm~ . It acts on a protein's nonterminal bonds which are adjacent to an aliphatic amino acid. Neutrophil elastase is of particular interest because it has the broadest spectrum of activity against natural connective tissue substrates. In particular, the elastase of the granulocyte is important because, as described above, granulocytes participate in acute infl~mm~tion and in acute exacerbation of chronic forms of infl~mm~tion which characterize many clinically important infl~mm~tory diseases.
Proteases may be inactivated by inhibitors which block the active site of the enzyme by binding tightly thereto. Naturally occurring protease inhibitors form part of the control or defense mech~ni~m~ that are crucial to the well-being of an org~ni~m. Without these control mech~ni~m~, the proteases would destroy any protein within reach. The naturally occurring enzyme inhibitors have been shown to have a~lo~riate configurations which allow them to bind tightly to the enzyme. This configuration is part of the reason that inhibitors bind to the enzyme so tightly (see Stroud, "A Family of Protein-Cutting Proteins" Sci. Am., July 1974, pp. 74-88). For example, one of the natural inhibitors, a1-Antitrypsin, is a glycoprotein contained in hllm~n serum that has a wide inhibitory spectrum covering, among other enzymes, elastase both from the pancreas and the PMN.
This inhibitor is hydrolyzed by the proteases to form a stable acyl enzyme in which the active site is no longer available. Marked reduction in serum al-antitrypsin, either genetic or due to oxidants, has been associated with pulmonary emphysema which is a disease characterized by a progressive loss of lung elasticity and resulting respiratory difficulty. It has been reported that this loss of lung elasticity is caused by the progressive, uncontrolled proteolysis or destruction of the structure of lung tissue by proteases such as elastase released from leukocytes. J. C. Powers, TIBS, 21 l (1976).
Rheumatoid arthritis is characterized by a progressive destruction of articular cartilage both on the free surface bordering the joint space and at the erosion front built up by synovial tissue toward the cartilage. This destruction process, in turn, is attributed to the protein-cutting enzyme elastase which is a neutral protease present in ~21 84385 human granulocytes. This conclusion has been supported by the following observations:
(1) Recent histochemical investigations showed the 5 accumulation of granulocytes at the cartilage/pannus junction in rheumatoid arthritis; and (2) a recent investigation of mechanical behavior of cartilage in response to attack by purified elastase demonstrated the direct participation of granulocyte enzymes, especially elastase, in 0 rheumatoid cartilage destruction. H. Menninger et al., in Biological Functions of Proteinases, H. Holzer and H. Tschesche, eds. Springer-Verlag, Berlin, Heidelberg, New York, pp. 196-206, 1979.
(F)-actin shortening proteins including gelsolin, plasma gelsolin (brevin), vitamin D binding protein (DBP), villin, fr~gmin, and 15 severin, and their use in shortening (F)-actin and treating (F)-actin mediated diseases is described in U.S. 5,260,224, issued to Stossel et al., on November 9, 1993, which patent is hereby incorporated by reference.
As discussed therein, Gelsolin has been cloned 20 (Kwiatkowski, D. J. et al., Nature, 323:455-458 (1986); Kwiatkowski, D. J. et al., J. Cell Biol., 106:375-384 (1988)) and fragments of the native protein which retain the ability to bind actin have been identified (Bryan, J., J. Cell Biol., 106:1553-1562 (1988); Yin, H. L. et al., J. Cell Biol., 107:465a (1988), abst. no. 2616); Kwiatkowski, D. J. et al., J.
25 Cell Biol., 108:1717-1726 (1989); Way, M. et al., J. Cell Biol., 109:593-605 (1989)).
Gelsolin's primary function in the plasma is to sever actin filaments. If gelsolin is present in excess of actin, only gelsolin-actin complexes result; if actin is in excess, there are free actin oligomers and 30 gelsolin-actin complexes. The actin severing occurs by way of a non-proteolytic cleavage of the noncovalent bond between adjacent actin molecules.
Efficacious levels of actin-binding compounds may be 7~11mini~tered so as to provide therapeutic benefits against the secondary toxic effects of excessive extracellular actin. By "efficacious levels" of - 21843~S

actin-binding compounds is meant levels in which the toxic effects of free extracellular actin are, at a ...i.lil.l-llll, ameliorated. By "excessive"
extracellular actin is meant an amount of extracellular actin which exceeds the ability of the plasma l~roteills to bind and clear the actin from extracellular fluids without secondary tissue (l~m~ge or toxic effects. By "secondary" tissue ~l~m~ge or toxic effects is meant the tissue damage or toxic effects which occur to otherwise healthy tissues, organs, and the cells therein, due to the presence of excessive extracellular actin in the plasma, usually as a result of a "primary"
tissue injury elsewhere in the body.
Stossel et al., recently presented a paper and abstract entitled Filamentous (F)-Actin Is Abundant In Cf Sputum, And F-Actin-Shortening Proteins Dimini.~h Sputum Viscosity In Vitro. Stossel et al., found actin in CF s~ululll by i~ oblotting, and CF s~u~um samples accelerated the nucleation of monumeric actin, consistent with the presence of F-actin. Stossel et al., speculated that F-actin therefore might contribute importantly to the mechanics of CF ~ululn. In a test model F-actin severing proteins in~ eously and substoichio-metrically shorten actin filaments, and nM concentrations of one such protein, hllm~n plasma gelsolin, rapidly reduced the viscosity of CF
sputum by up to 70% in a concentration-dependent manner. Gelsolin also ~limini~hed the elastic modulus of CF sputum by 50% in shear deformation over a frequency range of 0.001-10 Hz. Over ten times more bovine pancreatic DNAse I (which binds actin subunits in addition to its DNA hydrolyzing activity) was required to ~limini~h CF S~UlUlll viscosity by the same extent as gelsolin.
More recently, Vasconcellos et al., have reported that concentrations of 100 to 500 nM, gelsolin, purified from human plasma, rapidly tlimini~hed the viscosity of spulul-- form cystic fibrosis patients an average of 64% from 332~199 Pa-s over a 60 mimlte incubation period. On the other hand, at a concentration of 250 nM, bovine pancreatic DNase I and Gc golbulin had no effect on viscosity in this time period, however, they did appear to enhance gelsolin activity. See Science, Vol. 263, pp 969-971 (February 18, 1994).

BRIEF DESCRIPTION OF THE INVENTION
This invention relates to ph~rm~ceutical compositions and methods for the treatment of lung disease, and in particular Cystic Fibrosis comprising the a non-toxic effective amount of an elastase inhibitor such as the compounds of Formula (I), Ra R O~ ~ R
R~ ~ 4 ~N Rb O CONHCH ~ R2 ~\~

a non-toxic effective amount of a (F)-actin shortening protein, such as 5 gelsolin, and a pharmaceutically acceptable carrier.

DETAILED DESCRIPTION OF THE INVENTION
This invention relates to ph~m~.eutical compositions and methods for the treatment of lung disease, and in particular Cystic 20 Fibrosis comprising the a non-toxic effective amount of an elastase inhibitor such as the compounds of Formula (I), Ra 1 o~/

CONHCH ~~ R2 3 a non-toxic effective amount of a (F)-actin shortening protein, such as gelsolin, and a pharmaceutically acceptable carrier.
With regard to the compounds of Formula I, said compounds being further detailed in the Schemes, Fx~mples and Claims, the instant invention is more particularly directed to the compounds of the 218438~

Ra R~ ~\
~N Rb CONHCH ~ R2 ~J

o and pharmaceutically acceptable salts thereof wherein:
R is C1 6alkyl;
Rl is C1 6alkyl or C1 6alkoxy-C1 6alkyl;
Mis (1) hydrogen, (2) Cl 6alkyl, (3) hydroxy C1 6alkyl, (4) halo C1 6alkyl, (S) C2 6alkenyl, or (6) C1 -6alkoxy-cl -6alkYl;
Ra and Rb are each individually (1) hydrogen, (2) C1 6alkyl, (3) halo, (4) carboxy, (S) C1 6alkoxy, (6) phenyl, (7) C1 6alkylcarbonyl, (8) di-(C1 6alkyl)amino, (9) hydroxy;
3 0 R2 and R3 are each independently (1) hydrogen, (2) C1 6alkyl, (3) halo, (4) carboxy, (5) C1 6alkoxy, (6) phenyl, (7) C1 6alkylc~1,v.lyl, (g) ~oC2 3alkyIoxy carbonyl wherein the am~no is optd~nally mono or di sub~ti~te~
~1-6slkyl, (9) ~oC2 3al1yl~milw c~l,~lyl wll~ the amino is opnonally m~no or di ~ul~s~ te~
C1 6alkyl, (10) hydroxy, (11) ~mino~rbonyI whe~ein the amino is op~o~lly mono or d~ s~bsti~lteA uli~ Cl 6alkyl, (12) hydroxyme~yl, (13) ~m;nc,c~onyloxy Cl.3a~floxy wL.~ . ~e amino is Opei~n~l1y mono ar di substitllt~A with 1~ C~ cyl, (1~) cyano, (15) mo~ho~ o&~l~o~ phenyl~
(16) amino wherein ~c ~m;no is op~onally mono or di sub~it~lted wi~ C1 6alkyl, wi~h thc prolriso ~at R2 a~d R3 may be joined cl to fo~m a methylr~ liosy group or a furan ring, (1~) morphol;.loc~ yl;
;~ R?
R" is(a) a-~,-Y-N~ , or RE~

(b) Q-C-OR" where R,~ i5 carboxy G~ ~; alky!.

2~

WO 95t24207 2 1~ 4 3 8 5 PCT/US95/02938 benzyloxycarbonylCl 3alkyl, or t-butoxycarbonylC1 3alkyl, wherem Q is a covalent bond or wherein Rs and R6 are each individually Cl 3alkyl or hydrogen, / R12 \ ~10 Y is -N- C n--C--or / ~12 \ /R10 o I n-C--\ H /
or a covalent bond;

R12 is hydrogen or Cl 3alkyl;
R7 and R8 are each individually (a) hydrogen, (b) C1 6alkyl, (c) Cl 6alkyloxy C2 3alkyl, (d) hydroxy C2 6alkyl, (e) polyhydroxyC2 6alkyl, (f) carboxamido C1 -6alkYl, (g) polyacyloxyC2 6alkyl, (h) C1 6alkanoyl, (i) substituted phenyl or phenyl C1 6alkyl, wherein the substituent is X1 as defined immediately below, (j) C2 6alkenyl, (k) C6 10cycloalkenyl, o (l) heteroaryl C1 6alkyl wherein the hetero aryl includes pyridinyl, imidazolyl, triazolyl, benzylimidazolyl, and furyl, (m) carboxy C1 6alkyl, (n) carbo C1 6alkoxy Cl 3alkyl, (O) phenylsulfonyl, (p) Cl 6alkylsulfonyl, (q) benzyloxy, (r) morpholinyl C1 3alkylsulfonyl, (s) tetrahydropyranyl, (t) aminoC1 3alkylsulfonyl wherein the amino is optionally mono or di substituted with Cl 6alkyl, (u) aminocarbonyl wherein the amino is optionally mono or di substituted with C1 6alkyl, (V) aminocarbonyloxyC2 6alkyl wherein the amino is optionally mono or di substituted with Cl -6alkyl, (w) azabicyclo of 7 to 12 atoms, (x) di Cl 3alkylamino C2 6alkyl wherein the amino is optionally mono or di substituted with Cl 6alkyl, (y) bicycloalkyl of 7 to 12 atoms, (z) C3 10cycloalkyl optionally substituted with Cl 6alkyl, (aa) pyrazolidinyl, 21843~S

(bb) substituted piperidinyl or prrolidinyl wherein the substituent is hydrogen, C1 3alkyl, hydroxyCl 3alkylbenzyl, carboxamido or amino wherein the amino is optionally mono or di substituted with C1 6alkyl, (cc) substituted pyrrolidinyl wherein the substituent is carboxamido or amino wherein the amino is optionally mono or di substituted with C
o 6alkyl, (dd) pyrimidinyl, (ee) N-cyano-N'-phenylamidino, (ff) phosphonoCl -6alkyl, or (gg) a-C1 3alkyl benzyl or mono or di substituted benzyl or mono or di substituted pyridyl-methyl, wherein the substituents are X1 and x2 wherein Xl is (1) hydrogen, (2) halo, (3) Cl 6alkyl, (4) halo-C1 6alkyl, (5) C2 6alkenyl, (6) hydroxy-C1 6alkyl, (7) Cl 6alkylcarbonyl, (8) C1 6alkylcarbonylamino, (9) CN, (10) CF3, (11 ) CH30, (12) amino wherein the amino is optionally mono or di substituted with Cl 6alkyl, (13) carboxy, or (14) phenylsulfonylaminocarbonyl;
X2 is hydrogen, halo or C1 6alkyl;

n is 1, 2, 3, 4 or 5;

Rg is selected from hydrogen, Cl 4 alkyl, and C1 3alkoxyCl 3alkyl; orphenyl, phenyl Cl 3alkyl, pyridyl, and pyridyl C1 3alkyl;
R1o and R1 1 are each independently selected from hydrogen, C1 4alkyl, and C1 3alkoxy C1 3alkyl, or aryl as defined above, or are together O=; or wherein R7 and R8 are joined together to form mono or di substituted ring of 4, 5, 6, or 7 atoms or 7 to 12 atoms such as ( 1 ) piperidinyl or homopiperdinyl, (2) piperazinyl, (3) morpholinyl, thiomorpholinyl or 1,1-dioxo-4-thiomorpholinyl, (4) pyrroylidinyl, (S) pyrryl, (6) imidazolyl, 2 0 (7) triazolyl, (8) saturated azabicyclo of 7 to 12 atoms, (9) azaspiro having 3 to 9 carbon atoms, said ring being saturated, (10) tetrazolyl, (1 l) pyrazolidinyl, (12) dihydodimethoxyisoquinolyl, (13) azetidinyl, or (14) diazabicyclo ring of 7-12 atoms, wherein the substituents are each selected from the group consisting of hydrogen and C1 3alkyl, benzyloxycarbonyl, carboxy, phenyl C1 3alkyl amino carbonyl, pyrrolidinylmethyl, hydroxy C1 3alkyl, C1 6alkyloxy, Cl 4alkyloxy carbonyl, aminocarbonyl wherein the amino is optionally mono or di substituted with C1 6alkyl, and oxo; or -N(R7)R8 may be an amino acid residue including natural amino acids such as lysine; or WO 95/24207 PCT/US9~/02938 218 138~

R8 and Rg are joined together to form a mono or di substituted saturated monocyclic ring of 6 to 7 atoms and having two hetero atoms which are the nitrogens to which R8 and Rg are attached; said rings to include piperazinyl and homopiperazinyl; or Rg and Rlo are joined together to form a mono or di substituted monocyclic saturated ring of S to 7 atoms and having one hetero atom which is the nitrogen to which Rg is attached; or wherein Rg and R12 are joined together to form a mono or di o substituted saturated monocyclic ring of 5, 6, or 7 atoms, said ring having one hetero atom which is the nitrogen to which Rg is attached;
or wherein R1o and R12 are joined together to form a mono or di substituted saturated monocyclic ring of 5, 6, or 7 carbon atoms; or wherein R8 and R1 1 are joined together to form a mono or di substituted saturated monocyclic ring of 5, 6, or 7 atoms, said ring having one hetero atom which is the nitrogen to which R8 is attached;
and the substituents are independently selected from Hydrogen and C1 3alkyl.
As appreciated by those of skill in the art the term "alkyl"
such as in C1 6alkyl, includes, methyl, ethyl, propyl, butyl, pentyl, and hexyl, and where a~ro~riate, branched chained forms including isopropyl and tert-butyl.
As may also be appreciated by those of skill in the art, the (-CR12-)n spacer in definition Y, may, in the alternative be placed to the right of CR1oR11-~ R7 As may also be appreciated, the group -N, ,R7 may also be oxidized to the corresponding oxide -N ~ ~ O

In one Class the instant invention is directed to the compounds of the Formula (I) Ra R1 0~/~
R~ ~ R4 ~N Rb CONHCH ~ R2 ~\~

o and pharmaceutically acceptable salts thereof wherein:
R is C1 6alkyl;
R1 is C1 6alkyl or C1 6alkoxy-Cl 6alkyl;
Mis ( 1 ) hydrogen, (2) C1 6alkyl, (3) hydroxy C1 6alkyl, (4) halo C1 6alkyl, (5) C2 6alkenyl, or (6) C1 6alkoxy-C1 6alkyl;
Ra is (1) hydrogen, (2) C1 6alkyl, (3) halo, (4) carboxy, (5) C1 6alkoxy, (6) phenyl, (7) C1 6alkylcarbonyl, (8) amino wherein the amino is optionally mono or di substituted with C1 6alkyl;
3 0 Rb is hydrogen, or C1 -6alkyl, R2 and R3 are each independently (1) hydrogen, (2) C1 6alkyl, (3) halo, (4) carboxy, WO 95/24207 2 1 8 ~ ~ 8 5 PCT/US95/02938 (5) Cl_6alkoxy, (6) phenyl, (7) Cl 6alkylcarbonyl, (8) amino wherein the amino is optionally mono or di substituted with C1 6alkyl, or with the proviso that R2 and R3 may be joined together to form a methylenedioxy group or a furan ring;
o 0 7 Il ~R
R4 is (a)~Q~c~Y-N~R , or ll (b) -Q-C-ORX where Rx is carboxy-C~ 6alkyl, benzyloxycarbonylC1 3alkyl, or t-butoxycarbonylCl 3aLkyl, wherem Q is a covalent bond or I

30 wherein R5 and R6 are each individually C1 3alkyl or hydrogen Y is / ~12 \ ~(10 g \ / R "

/ ~12 \ / 10 o C ~tC--\ H / R~

or a covalent bond;

R12 is hydrogen or Cl 3aLkyl;
R7 and R8 are each individually (a) hydrogen, (b) C1 6alkyl, (c) C1 6alkyloxy C2 3alkyl, (d) hydroxy C2 6alkyl, (e) carboxamido Cl -6alkYl, (f) C1 6alkanoyl, (g) substituted phenyl or phenyl Cl -6alkyl wherein the substituents are Xl, and X2, (h) C2 6alkenyl, (i) C6 10cycloalkenyl, (j) heteroaryl C1 6aLkyl wherein the hetero aryl includes pyridinyl, imidazolyl, triazolyl, benzylimidazolyl, and furyl, (k) carboxy C1 6alkyl, (1) Cl 6alkylsulfonyl, (m) carboCl 6alkyloxyC2 3alkyl, (n) morpholinyl C1 3alkylsulfonyl, (o) aminoC1 3alkylsulfonyl wherein the amino is - 218438~

optionally mono or di substituted with Cl -6alkyl, (p) aminocarbonyl wherein the amino is optionally mono or di substituted with Cl 6alkyl, (q) aminocarbonyloxyCl 6alkyl wherein the amino is optionally mono or di substituted with C1 6alkyl, (r) di C1 3alkylamino C1 6alkyl wherein the amino is optionally mono or di substituted with Cl -6alkyl, (s) pyrazolidinyl, (t) substituted piperidinyl as defined above, (u) substituted pyrrolidinyl as defined above, (v) pyrimidinyl, (w) benzyloxy, (x) C3 10cycloalkyl, (z) a-C1 3alkyl benzyl or mono or di substituted benzyl or mono or di substituted pyridylmethyl, wherein the substituents are X
and X2, wherein Xl is (1) hydrogen, (2) halo, (3) Cl 6alkyl, (4) halo-C1 6alkyl, (5) C2 6alkenyl, (6) hydroxy-C1 6alkyl, 0 (7) C1 6alkylcarbonyl, (8) C1 6alkylcarbonylamino, (9) di-Cl 3alkylamino, or (10) carboxy;
X2 is hydrogen, halo or C1-6alkYI;

n is 1 , 2, 3, 4 or 5;

R9 is selected from hydrogen, C1 4 alkyl, and C1 3alkoxyC1 3alkyl;
Rlo and R1 1 are each independently selected from hydrogen, C1 4alkyl, and C1 3alkoxy C1 3alkyl; or wherein R7 and R8 are joined together to form mono or di substituted o ring of 4, 5, 6, or 7 atoms such as ( 1 ) plperldinyl, (2) piperazinyl, (3) morpholinyl, (4) pyrroylidinyl, (5) pyrryl, (6) imidazolyl, (7) triazolyl, (8) tetrazolyl, (9) pyrazolidinyl, (10) azetidinyl, wherein the substituents are each selected from the group consisting of hydrogen and C1 3alkyl, beniyloxycarbonyl, carboxy, phenyl C1 3alkyl amino carbonyl, pyrrolidinyl, methyl, hydroxy C1 3alkyl, C1 6alkyloxy, C1 4alkyloxy carbonyl, and oxo; or R8 and Rg are joined together to form a saturated ring of 5 to 7 atoms and having two hetero atoms; or Rg and R1o are joined together to form a saturated ring of 5 to 7 atoms and having one hetero atom; or wherein Rg and R12 are joined together to form a ring of 5, 6, or 7 atoms, said ring being saturated; or wherein R1o and R12 are joined together to form a ring of 5, 6, or 7 atoms, said ring being saturated;
or wherein R8 and R1 1 are joined together to form a ring of 5, 6, or 7 atoms, said ring being saturated and having one hetero atom.

-218 138~

In one subclass, the invention concerns compounds of Formula I
wherein R is C1 3alkyl;
Rl is Cl 3alkyl;
Mis (a) C 1 -6alkyl, or (b) C2-6alkenyl;
R2 is (a) hydrogen, (b) C1 6alkyl, or C1 6alkoxy, and R3 is hydrogen, or R2 and R3 are joined together to form a methylenedioxy 15group or a furan ring;
R5 and R6 are each individually hydrogen or C1 3alkyl;
R7 and R8 are each independently selected from (a) hydrogen, (b) C1 3alkyl, (c) C1 3alkoxy C2 3alkyl, (d) C3 7cycloalkyl, (e) hydroxyC2 3alkyl, (d) carbo C1 4alkyloxymethyl, (g) substituted benzyl wherein the substituents are X1 and X2 25wherein X1 is hydrogen and X2 is (1) hydrogen, (2) halo, or (3) C~1 3alkyl;

nis 1,2or3,and Rg, R1o and R1 1 are each independently selected from hydrogen, C1 4aIkyl, and C1 3alkoxy C1 3alkyl; or R7 and R8 are joined together to form a substituted ring selected from (a) piperidinyl, (b) piperazinyl, and (c) morpholinyl;
or R8 and Rg are joined together to form a ring of 6 to 7 atoms and having two hetero atoms;
Rg and Rlo are joined together to form a saturated ring of 5 to 7 atoms and having one hetero atom; or wherein Rg and R12 are joined together to form a ring of S, 6, or 7 atoms, said ring being saturated; or wherein Rlo and R12 are joined together to form a ring of S, 6, or 7 atoms, said ring being saturated;
or wherein R8 and Rl 1 are joined together to form a ring of S, 6, or 7 atoms, said ring being saturated and having one hetero atom.

In a narrower sub-class are the compounds wherein Q is a covalent bond;
R is methyl or ethyl;
Rl is methyl or ethyl;
- Mis (a) Cl 4alkyl, or (b) C2 3alkenyl;
R2 is (a) hydrogen, O (b) Cl 3alkyl, or Cl 3alkoxy, and R3 is hydrogen, or R2 and R3 are joined together to form a furan or dioxacyclopentane ring;
nis 1 or2;
Rg and Rlo are each independently selected from - 218~8~

(a) Cl 3alkyl, (b) Cl 3alkoxy Cl 3alkyl, (c) hydrogen, R7 and R8 are each independently selected from (a) Cl 3alkyl, (b) Cl 3alkoxy C2 3alkyl, (c) hydrogen, (d) hydroxyethyl, o (e) carboethoxymethyl, (f) cyclopropyl, or R7 and R8 are joined together to form a substituted ring selected from (a) piperidinyl, and (b) morpholinyl, or R8 and Rg are joined together to form a piperazine ring.

As is defined above, various rings are formed when R8, 20 Rg, R1o and R12 are joined. The following is a non-limiting description of some of the preferred rings that are formed when these various substituents are joined.

R~ and R9 are joined R,~ R" ~) ,R~

Rg and Rlo are joined s ~R7R~ 12~'~NR7R~ [~NR7R~

~<NR7Rs ~ R11 ~NJ 1' 12 ~N) ~N! R12 R 12 ~< R 11 1~ N R7R8 /~ N R7R8 ~N NR7R8 INJ ~NJ 12 W O 95/24207 PCTrUS95/02938 Rg and Rl~ are joined ~ ~ N R7R8 R10 R" R ~NR7R8 NR7RB ~ C~,O.RN"

Rlo R11 R10 R11 >~ NR7R8 ~ N R7R8 _ _ _ . _ _ .

2l 84385 Rlo and Rl ~ are joined PCNR7R8 5~ 1' Ç<R"

R[~R7R8 ~ ~ NRg ~ ~ NRg ~ ~ NRg ~R11 ' /~NR7R8 1~ NR7R8 ~ ~NRg ~ ~NRg WO 95/24207 2 18 4 ~ ~ ~ PCT/US95/02938 Rlo and Rl ~ are joined ;-- Rg I ~ RgN~

Rg RgN~ R7 R9!N~ "' ~) As discussed above, (F)-actin shortening proteins are effective in reducing the viscosity of sputum, particularly cystic Fibrosis SpUIulll. As shown by the Vasconcellos et al., reference cited above, liquefaction is not instantaneous. However, once a suff1cient degree of viscosity reduction is achieved, a patient can more easily expectorate or otherwise rid himself of excess sputum.
One problem that has not been appreciated is that liquefaction, considered alone, may affect the patient adversely. For example, viscosity reduction may provide the destructive proteases within the sputum greater access to lung and related tissues. Thus, the applicant has found it to be of surprising importance to inhibit these destructive proteases during the transition period from the onset of viscosity reduction until the ~l~u~unl is expectorated. Moreover, the 15 high proportion of protease destruction caused by the elastase in the sputum has also gone unappreciated.
The compounds of Formula I are surprisingly, highly active in viscous ~yulunl as well as liquified ~Ululll.
As a result of the above factors, treatment with applicants 2 composition is capable of returning a patent with lung disease to substantially normal lung function, as measured, for example, by FEV
(forced expiration volume). In particular, treatment with applicants composition is capable of retllrning a patent with lung disease to at least 60-75% of normal lung function, as measured, for example, by FEV1.
Moreover, treatment with applicants composition is capable of returning a patent with lung disease to 75-90% of normal lung function or greater, as measured, for example, by FEV1.
Accordingly, in one aspect the invention encompasses a method of treating a patient with a lung disease, comprising:
~(lmini.~tration to a patient in need of sputum viscosity reduction, a therapeutically effective non-toxic amount of an (F)-actin shortening protein and a therapeutically effective non-toxic amount of compound of Formula I as described herein.
Within this aspect the invention encompasses a method of treating a patient with a lung disease, comprising:

-- 218~8~

~tlmini~tration to a patient in need of sputum viscosity reduction, a therapeutically effective non-toxic amount of an (F)-actin shortening protein and a therapeutically effective non-toxic amount of compound 5 of Formula I as described herein, said amounts effective to return the lung function of said patients to at least 60-75% of normal as measured by FEV1.
Within this class the invention encompasses a method of treating a patient with a lung disease, comprising:
atlmini.~tration to a patient in need of sputum viscosity reduction, a therapeutically effective non-toxic amount of an (F)-actin shortening protein and a therapeutically effective non-toxic amount of compound of Formula I as described herein, said amounts effective to return the lung function of said patients to 75 to 90% of normal as measured by FEV 1 .
The compounds of the invention are prepared by known methods or are prepared among other methods by the following representative schemes. For example, methods for making such compounds are disclosed in EP O 337 549, published October 18, 1989, 20 which is hereby incorporated by reference.
This invention also relates to a method of treating infl~mm~tion in patients using a compound of Formula (I), particularly a preferred compound as the active constituent.
It has been found that the following compound are effective 2 inhibitors of the proteolytic function of human neutrophil elastase as shown below in Table 1 to 10.

WO 95/24207 PCT/US9StO2938 Et "" ~11 O A

NH

Pr No. A Kobs/rIl -cH2cH2N(cH3)2 1,566,000 2 -CH2CO2H 1,667,000 3 -CH2-C(O)N(CH2CH2OH)2 3,428,000 4 -CH2-C(O)N(CH3)CH2C(O)NH2 4,293,000 -CH2C(O)NH-C(CH2OH)3 4,448,000 6 -cH2c(o)N(cH3)2 2,997,000 7 -CH2CH2N(CH3)Ac 1,558,000 8 -CH2C(O)-Pro-OCH2Ph 12,501,000 9 -CH2C(O)-Pro-OH 1,571,000 -CH(CH3)CO2CH2Ph 2,891,000 11 -CH(CH3)CO2H 1,132,000 12 -CH(CH3)C(O)N(Et)2 2,815,000 13 -CH(CH3)CH2N(CH3)2 2,472,000 14 -CH2CH2CH2N(CH3)2 2,855,000 -CH2CH2N(O)(CH3)2 2,162,000 16 -CH2CH2N(Et)2 2,291,000 17 -CH2CH2(4-morpholinyl) 4,733,000 18 -CH2CH2CH2CH2N(CH3)2 1,934,000 19 -CH2C(O)-Pro-NHCH2Ph 4,956,000 -CH2C(CH3)2N(CH3)2 1,470,000 21 -CH2CH2N(i-Pr)2 1,671,000 22 -CH2CH2(4-carbobenzyloxy- 1 -piperazinyl) 4,115,000 218~8~

No. A Kobs/rIl 23 -CH2CH2N(n-Bu)2 992,000 24 -cH2cH2cH2cH2cH2cH2N(cH3)2 1,988,000 -CH2CH2(1 -piperazinyl) 1,709,000 26 -CH2CH2(4-methyl-1-piperazinyl) 4,685,000 27 -CH2CH2(4-acetyl- 1 -piperazinyl) 3,262,000 28 -CH2CH2N(Ph)2 188,000 29 -CH2CH2N(CH2CH=CH2)2 891,000 -cH2cH(ph)N(cH3)2 656,000 31 -CH2CH2N(CH3)CH2Ph 1,180,000 Et `~" ~CH2-C-O-A

\~ CH3 - NH

Pr No. A Kobs/lIl 32 -CH2CH2N(CH3)2 1,993,000 33 -CH2CH2CH2N(CH3)2 1,151,000 34 -CH2CH2N(Et)2 1,339,000 -CH2CH2-(4-morpholinyl) 1,725,000 36 -CH(CH3)CH2N(CH3)2 1,688,000 37 -CH2-C(CH3)2N(CH3)2 2,100,000 38 -CH2CO2H 1,008,000 39 -CH2CH2N(CH3)CH2Ph 751,000 WO 9~/24207 PCT/US95/02938 Et ` ~
o.~N O CH2-C-A

NH

o Pr No. A Kobs/rIl -N(CH2CH2OH)2 1,241,000 41 4-methyl-1-piperazinyl 974,000 42 4-morpholinyl 1,088,000 43 -NHcH2cH2N(cH3)2 1,211,000 44 -N(CH3)CH2CH2N(CH3)2 1,243,000 -NHCH2CH2CH2N(CH3)2 1,118,000 46 -NHCH2CH2-(4-pyridyl) 2,254,000 47 -NHCH2CO2H 876,000 48 -NHCH(CH3)CO2H 676,000 49 -NHcH2c(o)N(cH2cH2oH)2 1,295,000 -N(CH3)CH2CO2H 989,000 51 -NHCH(CH3)C(O)N(CH2cH2OH)2 939,000 52 -N(cH3)cH2c(o)N(cH2cH2oH)2 273,000 53 -N(CH3)CH2CH2-(4-morpholinyl) 2,511,000 54 -N(CH3)CH2CH2N(cH2cH2OcH3)2 1,388,000 SS -N(CH3)CH2CH2N(Et)2 1,316,000 56 -N(CH3)CH2CH2CH2N(CH3)2 1,047,000 57 -NHCH2CH(CH3)N(CH3)2 1,344,000 58 -N(CH3)CH2CH2N(i-Pr)2 1,634,000 59 -N(n-pr)2 1,144,000 WO 95/24207 2 1 8 4 3 8 ~ PCT/US95/02938 No. A Kobs/rIl -N(Et)2 1,079,000 61 3-chloroanilino- 733,000 62 3-methoxyanilino- l ,621,000 63 4-fluoroanilino-64 -N(CH3)CH2CH2CH2CO2H 917,000 -N(CH3)CH2CH2CH2C(O)NHSO2Ph 1,335,000 66 -N(CH3)CH2CH2CH2N(cH3)cH2Ph 1,355,000 67 -N(CH3)2 942,000 o 68 -N(CH3)CH2Ph 1,897,000 69 -N(CH3)CH2CH2N(CH3)CH2Ph 2,792,000 -NH-O-CH2Ph 2,371,000 71 -N(CH3)(4-carboxyphenyl) 1,508,000 72 -N(CH3)(4-benzenesulfonylaminocarbonyl-) phenyl) 3,284,000 Et~`~" ~C-A

\~ CH3 NH~
2s Pr No. A Kobs/rIl 73 -NHCH2CH2N(CH3)2 968,000 74 -NH-CH2CO2H 1,434,000 -N(CH3)CH2CH2N(CH3)2 1,916,000 76 -N(Et)CH2CH2N(CH3)2 1,436,000 77 -NHCH2CH2N(Et)2 1,187,000 78 -NHCH2CH2-(4-morpholinyl) 1,841,000 79 -N(CH3)CH2CH2-(4-morpholinyl) 2,118,000 21 8~385 No. A Kobs/rIl -N(CH3)CH2CH2N(CH2CH2OCH3)2 2,078,000 81 -N(CH3)CH2CH2N(Et)2 2,191,000 82 -N(Ph)CH2CH2N(CH3)2 2,504,000 83 -N(cH3)cH2cH2cH2N(cH3)2 1,797,000 84 -NHCH2CH2N(i-Pr)2 2,100,000 -N(CH3)CH2CH2N(O)(CH3)2 1,589,000 86 -N(CH3)CH2CH2N(i-Pr)2 2,449,000 87 -NH-SO2CH2CH2-(4-morpholinyl) 775,000 88 -NH-SO2CH2CH2N(CH3)2 788,000 89 -NHCH2CH2-(4-imidazolyl) 2,092,000 -NHCH2CH2-(1 -piperidinyl) 941,000 91 -N(CH3)CH2CH2-(1-piperidinyl) 892,000 92 -N(CH3)CH2CH2NHCH3 1,453,000 93 -N(CH3)CH2CH2N(CH3)Ac 1,960,000 94 -NHCH2CH2-(1-pyrrolidinyl) 1,239,000 -N(CH3)CH2CH2-(1 -pyrrolidinyl) 1,005,000 96 -NHCH2CH2-(1 H- 1,2,4-triazol- 1 -yl) 1,397,000 97 -NH-CH2CH2-(1-imidazolyl) 1,070,000 98 -NH-CH2CH2-(3-azabicyclo-[3.2.2-non-3-yl) 3,043,000 99 -NH-CH2CH2-(3-azaspiro[5.5]-undec-3-yl) 2,583,000 100 -NH-CH2CH2-(2H-tetrazol-2-yl) 2,006,000 101 -NH-CH2CH2-(lH-tetrazol-1 -yl) 2,053,000 102 -NHCH2C(O)-Pro-NHCH2Ph 2,747,000 103 -N(CH3)CH2CH2-(3-azabicyclo-[3.2.2]non-3-yl) 2,996,000 104 -N(CH3)CH2CH2-(4-imidazolyl) 2,389,000 105 -N(CH3)CH2CH2N(CH3)Ac 2,398,000 106 -N(CH3)CH2CH2N(CH3)C(O)NHCH3 2,486,000 107 -N(CH3)CH2CH2N(CH3)SO2CH3 2,530,000 108 -N(CH3)CH2CH2(3-azabicyclo-[3.2.2]non-3-yl) 2,953,000 109 -NHCH2CH2-(1,1 -dioxo-4-thiamorpholinyl)1,275,000 110 4-dimethylaminobenzylamino 5,598,000 218~

No. A Kobs/rIl 111 3-dimethylaminoanilino 2,286,000 112 -N(CH3)CH2CH2-(1,1 -dioxo-4-thia-morpholinyl) 1,596,000 113 4-dimethylaminoanilino 2,591,000 114 -NHCH2CH2-(l-benzyl-lH-imidazol-2-yl) 3,853,000 115 -N(CH3)CH2CH2(2-pyridyl) 2,272,000 116 -N(CH3)(1-azabicyclo[2.2.2]oct-3-yl 3,480,000 117 -NHCH2CH2(4-benzyloxycarbonyl- 1 -piperazinyl) 6,231,000 118 1,2-diethylpyrazolidin-4-ylamino 1,001,000 119 2-(1-S-pyrrolidinylmethyl)-1-pyrrolidinyl 2,692,000 120 -NHCH2CH2(4-hydroxy- 1 -piperidinyl) 1,728,000 121 -NHCH2CH2(1 -homopiperidinyl) 2,069,000 122 -N(CH3)CH2CH2(1-homopiperidinyl) 2,899,000 123 -NHCH2CH2(3-hydroxy- 1 -piperidinyl)1,534,000 124 -N(CH3)CH2CH2(3-hydroxy- 1 -piperidinyl) 1,963,000 125 -N(CH3)CH2CH2N(CH3)CH2Ph 2,054,000 126 -N(CH3)CH2CH2(4-benzyloxy-1-piperidinyl) 3,476,000 127 -N(n-Pr)2 128 -N(Et)2 1,454,000 129 -N(CH3)CH2CH2(4-hydroxy- 1 -piperidinyl) 1,994,000 130 -N(CH3)CH2CH2(4-oxo- 1 -piperidinyl)2,297,000 131 -NHCH2CH2(3-hydroxy-1-pyrrolidinyl) 1,111,000 132 -N(Et)CH2CH2(1-piperidinyl) 1,244,000 133 -N(CH2Ph)CH2CH2(1-piperidinyl) 1,521,000 134 4-fluoroanilino- 724,000 135 3-chloroanilino- 201,000 136 3-methoxyanilino 3 137 -N(CH2Ph)CH2CH2N(CH3)2 1,380,000 138 -N(CH3)CH2CH2(3-hydroxy-l-pyrrolidinyl)960,000 139 -N(3-picolyl)CH2CH2(1 -piperidinyl)1,189,000 140 -NHCH(CH3)CH2CH2CH2N(Et)2 1,361,000 No. A Kobs/rIl 141 -NHCH2CH2(2-S-hydroxymethyl-1-pyrrolidinyl 1,507,000 142 -N(CH3)CH2CH2(4-t-butoxycarbonyl- 1 -piperazinyl) 3,471,000 143 -N[CH2CH2N(CH3)2]2 1,878,000 144 -N[CH2CH2N(Et)2]2 1,508,000 145 -N(CH3)CH2CH2N(CH3)(3-picolyl) 2,877,000 146 3,5-dimethyl-1 -piperazinyl 1,518,000 147 -N(CH3)CH2CH2N(O)(CH3)CH2Ph 2,493,000 148 -N(CH3)CH2CH2N(CH3)(4-picolyl) 2,389,000 149 2-S-(N-benzyl-N-methylaminomethyl)-1-pyrrolidinyl 3,268,000 150 -N(CH3)CH2CH2N(CH3)(2-picolyl) 2,165,000 151 -N(CH3)CH2CH2(1-piperazinyl) 1,191,000 152 1-homopiperazinyl 1,951,000 153 -N(CH3)CH2CH2N(CH3)cH2cH2Ph 2,797,000 154 2-(1 -R-pyrrolidinylmethyl)- 1 -pyrrolidinyl 1,666,000 155 4-benzyl-1-homopiperazinyl 1,979,000 156 -N(cH3)cH2-[cH(oH)]4cH2oH 1,198,000 157 -N(CH3)CH2-[CH(OAc)]4CH2OAc 1,171,000 158 -N(CH3)CH2CH2N(CH3)(1-Naphthalenyl-methyl) 1,075,000 159 -N(CH3)CH2CH2N(CH3)(2-Naphthalenyl-methyl) 1,337,000 160 -N(CH3)CH2CH2N(CH3)CH(CH3)Ph 1,569,000 161 -N(CH3)CH2CH2N(CH2Ph)2 1,021,000 162 1-ethyl-3-piperidinylamino 949,000 163 -N(CH3)CH2CH2N(CH3)(2-furfuryl) 1,818,000 3 164 -N(CH3)CH2CH2CH2CO2H 1,064,000 165 -N(CH3)CH2CH2CH2C(O)NHSO2Ph 1,550,000 166 -N(CH3)CH2CH2N(CH3)CH2CH=CH2 1,359,000 167 -N(CH3)CH2CH2CH2N(CH3)CH2Ph 1,293,000 168 -N(cH3)-(cH2)6-N(cH3)cH2ph 2,157,000 218~3~

No. A Kobs/rll 169 -N(CH3)CH2CH2OH 1,457,000 170 -N(cH3)cH2cH2oc(o)N(cH3)2 1,518,000 171 -N(CH3)CH2CH2N(CH3)CH2CO2-t-Bu 1,831,000 172 -N(CH3)(1-ethyl-3-piperidinyl) 1,545,000 173 -N(CH3)CH2CH2N(CH3)(tetrahydro-2H-pyran-2-yl-methyl) 2,943,000 174 2,2,6,6-tetramethylpiperidin-4-ylamino869,000 175 -N(CH3)(4-carboxyphenyl) 1,055,000 176 -N(CH3)(4-benzenesulfonylaminocarbonyl-phenyl 3,231,000 177 -N(CH3)CH2CH2N(CH3)(4-cyanobenzyl)2,201,000 178 -N(CH3)CH2CH2N(CH3)(4-methylbenzyl)1,870,000 179 -N(CH3)CH2CH2N(CH3)(3-cyanobenzyl)2,448,000 180 -N(CH3)CH2CH2N(CH3)(4-trifluoromethyl-benzyl 905,000 181 -N(CH3)CH2CH2N(CH3)(3-trifluoromethyl-benzyl 564,000 182 -N(CH3)CH2CH2(CH3)(4-fluorobenzyl)2,137,000 183 -NHCH(CH3)PH(O)OH 977,000 184 L-lysine (a-N) 755,000 185 -N(CH3)CH2CH2N(CH3)(cyclopropylmethyl) 1,787,000 186 -N(CH3)CH2CH(Ph)N(CH3)2 1,053,000 187 -N(CH3)2 1,749,000 188 -N(CH3)CH2Ph 1,837,000 189 -N(CH3)(1-benzyl-3-piperidinyl) 1,879,000 190 -NH-O-CH2Ph 1,797,000 191 -N(3 -picolyl)CH2CH2N(CH3)CH2PH 2,538,000 192 -N(CH3)CH2CH2N(CH3)(4-me~oxybenzyl)1,785,000 193 -N(4-picolyl)CH2CH2N(CH3)CH2Ph 2,243,000 194 -N(2-picolyl)CH2CH2N(CH3)CH2Ph 2,473,000 195 -N(CH3)CH2CH2N(CH3)(2,4-dimethylbenzyl) 1,119,000 No. A Kobs/rIl 196 -N(CH3)CH2CH2(2,6-dimethyl-4-morpholinyl) 1,530,000 197 -NH2 1,638,000 198 -NHCH3 1,825,000 199 4-morpholinyl 2,376,000 200 cis-2,6-dimethyl-4-morpholinyl 1,837,000 201 -NH-CH2CH2CH2CH3 2,460,000 202 -N(CH3)CH2CH2N(CH3)C(=N-CN)NHPh1,763,000 203 -N(CH3)CH2CH2N(CH3)(3-fluorobenzyl) 1,262,000 204 -N(CH3)CH2CH2N(CH3)(2-chlorobenzyl) 1,591,000 205 -N(CH3)CH2CH2N(CH3)(3-methoxybenzyl) 1,911,000 206 -N(CH3)CH2CH2N(CH3)(3,5-dimethoxy-benzyl) 1,735,000 207 3,4-dihydro-6,7-dimethoxy-2-(1 H)iso-quinolinyl 2,698,000 208 -N(CH3)(1-benzyl4-piperidinyl) 1,948,000 209 L-lysine (-N) 929,000 210 -N(CH3)CH2CH2N(CH3)(2-~d~m~ntyl)2,132,000 211 -N(CH3)(4-piperidinyl) 85,000 212 S-Methyl-2,5-diazabicyclo[2.2.1]hept-2-yl 860,000 213 -N(CH3)CH2CO2H 984,000 214 -N(CH3)CH2CH2CH2N(CH3)CH2CH3 1,099,000 215 -N(CH3)(1 -methyl-4-piperidinyl.)1,283,000 216 -N(CH3)(1 -propyl-4-piperidinyl)1,312,000 217 -N(CH3)(1-ethyl-4-piperidinyl) 1,422,000 218 -N(CH3)CH2CH(CH3)N(cH3)cH2Ph 2,123,000 219 -N(CH3)CH2CH(CH3)N(CH3)2 1,588,000 220 -N(CH3)CH2CH2N(CH3)(bicyclo[2.2.1]-hept-2-yl) 1,874,000 221 -N(CH3)CH2CH2NH(2-~m~ntyl) 3,010,000 222 -N(CH3)CH2CH2N(CH3)(6,6-dimethylbicyclo-[3.1.1]hept-2-yl 2,288,000 No. A Kobs/rIl 223 -N(CH3)CH2CH2N(CH3)(bicyclo[3.2.1]-oct-2-yl) 2,584,000 224 -NH(t-Bu) 225 -N(CH3)CH2CH2N(CH3)(1-cyclohexen-1-yl) 1,839,000 226 -N(cH3)cH2cH2NHc(cH3)2cH=cH2 1,309,000 227 2-S-carboxamido- 1 -pyrrolidinyl 931,000 228 2-hydroxymethyl- 1 -piperidinyl 50,000 229 3-dimethylamino-1-pyrrolidinyl 1,336,000 230 -N(CH3)CH2CH2N(CH3)(cyclohexylmethyl) 231 -N(CH3)CH2CH2N(CH2CH=CH2)C(CH3)2-CH=CH2 925,000 232 -N(CH3)CH2CH2N(CH3)(4-ethylcyclohexyl) 2,476,000 233 -N(CH3)CH2CH2N(CH3)(2-ethylcyclohexyl) 2,030,000 234 -N(CH3)CH2CH2N(CH3)(4-methylcyclohexyl) 2,166,000 235 -N(CH3)CH2CH2N(CH3)(cyclohexyl) 1,952,000 236 -N(CH3)CH2CH2N(CH3)CH2CO2H-TFA 31,000 237 -N(CH3)CH2CH2N(CH3)CH2C(O)N(CH3)2 2,679,000 238 3-dimethylamino- 1 -azetidinyl 239 1-diphenylmethyl-3-azetidinyl 240 -N(CH)CH2CH2N(CH3)(cyclohexylmethyl) 3,003,000 241 -NHCH2CH2N(Et)CH2CH2OCH3 1,090,000 E t ""` ~3\
o~N~O ll N N-R

~CH3 NH~

o Pr No. R Kobs/rIl 242 -CH3 1,700,000 243 4-fluorophenyl 7,486,000 244 3-chlorophenyl 2,453,000 245 phenyl 5,276,000 246 benzyl 5,171,000 247 H 1,100,000 248 i-Pr 2,392,000 249 i-Bu 2,476,000 250 -CH2CO2Et 1,571,000 251 -CH2CO2H 1,947,000 252 Et 2,324,000 253 Pr 1,768,000 254 2-pyrimidinyl 2,142,000 255 -CH2CH2OC(O)NHCH3 2,548,000 256 cyclopropyl 3,587,000 256a -CH2CH2OH 2,000,000 218~3~5 Et "` ~l ~ N (cH2)n-GA
O \~

NH~ [ >

o Pr No. n A Kobs/rIl 257 1 NH2 2,342,000 258 1 4-morpholinyl 1,785,000 259 1 -N(CH3)CH2CH2N(CH3)2 2,522,000 260 0 -N(CH3)CH2CH2N(CH3)2 3,317,000 261 0 -N(Et)2 3,207,000 262 0 -N(CH3)(n-Bu) 3,125,000 263 0 4-methyl-1-piperazinyl 3,805,000 264 0 -N(CH3)CH2CH2N(CH3)CH2Ph 3,427,000 265 0 4-cyclopropyl- 1 -piperazinyl 4,500,000 265a 0 1-piperazinyl 3,250,000 265c 0 4-(2-hydroxyethyl)- 1 -piperazinyl 4,800,000 265d 0 4-morpholinyl 3,700,000 Et ""` ~3` 1l .~ N~o (C H2)n-C-A

~R4 NH~`CO N O

o Pr \ J

No. n B4 A Kobs/~Il 266 1 H 4-morpholinyl 169,000 267 1 H -N(Et)2 33 268 1 H -N(CH3)CH2CH2N(CH3)2 142,000 269 1 CH3 NH2 637,000 270 1 CH3 N(Et)2 740'000 271 1 CH3 N(n-Pr)2 826,000 272 0 Et -N(CH3)CH2CH2N(CH3)2 2,423,000 273 0 Et -N(CH3)(n-Bu) 3,258,000 WO 95/24207 PCT/I~S95/02938 E ""` ~
N (CH2)n-GA
O' \f~O
N~ ~CRON~O

o Pr No. n R3 A Kobs/~Il 274 1 H NH2 430,000 275 1 H -N(CH3)CH2CH2N(CH3)2 290,000 276 1 H -OCH3 440,000 277 0 H -N(CH3)CH2CH2N(CH3)2 548,000 278 0 H -OCH2CH2N(CH3)2 135,000 279 0 H -N(Et)2 566,000 280 0 H 4-moIpholinyl 577,000 2 s Et "`~ ~l(CH2)n-C-A

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V V V V V V V V V
Z Z Z Z Z Z Z Z Z
T ~ ~I T
V V V V V V V V V
o o o V V V V V V ~ V o o V

~_ ~V ~ ~V ~ V V ~V ~ ~o V
d- d d I I I I I I I I d- I d V
~ Z~
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V V
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~ '' " o '' '' '' o o X ~ X V V

~L~
z x ~ x :c x ~ ~ x ~ O m _ ~ o o o ~ o o ~ ~ ~ o o~ 1 ~ d- ~ ~0 r-- oo CJ~ O ~ C` ~') d-,_ oo oo oo x x x oo x oo cr~ ct~ C~ Cl~ cr~ ct~

o o o o o o o o o o o o ~ 8 o o ~ o Q
D 1~ Y 0~ ~C
O ~ O 00 ~ ~
_ _ _ _ _ _ G
5: ~ X
~ V
V ~
Z Z .~i Z Z Z Z Z
X
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I ~ ~ ~ ~ ~ X 5 V
X
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~:1 Z Z ~ Z Z Z Z Z

~ ~ ~l o ~ ~ v ~ ~

~l ~
:~ x o ~1oooooooo ol ~O 1~ x c~ O ~
~ ~ o~ o o o o 843a5 Et ~C-A

~,CH3 NH~

o Pr No. R~ B6 A Kobs/~II
304 CH3 CH3 -OCH2CH2N(CH3)2563,000 305 CH3 CH3 -OCH2CH2N(Et)2 749~000 306 CH3 CH3 -OCH2CH2N(i-Pr)2 612,000 307 CH3 CH3 -N(CH3)CH2CH2N(CH3)2 352,000 308 CH3 CH3 -N(cH3)cH2cH2N(Et)2 377,000 309 CH3 CH3 -N(Et)CH2CH2N(CH3)2 398,000 310 H OH -N(CH3)CH2CH2N(CH3)2 838,000 Enzyme Assays for the Inhibition of Human Polymorphonuclear Leukocyte Elastase Via Hydrolysis of N-t-Boc-alanyl-alanyl-prolyl~l~nine-p-nitroanilide (Boc-AAPAN) or N-t-Boc-alanyl-25 prolylvaline-p-nitro-anilide (Boc-AAPVN) Reagent:
0.05M TES (N-tris[hydroxymethyl]methyl-2-mino-ethanesulfonic acid) Buffer, pH 7.5.
0.2 mM Boc-AAPAN or Boc-AAPVN.
To prepare substrate, the solid was first dissolved in 10.0 30 ml DMSO. Buffer at pH 7.5 was then added to a final volume of 100 ml.
Crude extract of human polymorphonuclear leukocytes (PM:N) cont~inin~ elastase activity.

Inhibitors (azetidinones) to be tested dissolved in DMSO
just before use.
To 1.0 ml of 0.2 mM Boc-AAPAN in a cuvette, 0.01-0.1 5 ml of DMSO with or without inhibitor was added. After mixing, a measurement was taken at 410 m,u to detect any spontaneous hydrolysis due to presence of test compound. 0.05 ~illiliters of PMN extract was then added and the ~OD/min at 410 m~l was measured and recorded.
Beckman model 35 spectrophotometer was used.
Results were expressed to the nearest thousand Kobs/I
which is the second order rate constant in per mole per second for inactivation of the enzyme.
The elastase activity in the crude PMN extract may vary from one preparation to another. A control of each new batch is run, 15 and the volume added in the assay procedure is adjusted according to actlvity.
This invention also relates to a method of treating infl~mm~tion in patients using a compound of Formula (I), particularly a preferred compound as the active constituent.
It has been found that the compounds of Formula (I) are effective inhibitors of the proteolytic function of hllm~n neutrophil elastase.
Accordingly, the compounds of Formula (I), can be used to reduce infl~mm~tion and/or relieve pain in diseases such as emphysema, rheumatoid arthritis, osteoarthritis, gout, bronchial i~Lfl~mm~tion, chronic or acute bronchitis, cystic fibrosis, adult respiratory distress syndrome, atherosclerosis, sepsis, septicemia, shock, periodontitis, glomerular nephritis or nephosis, myocardial infarction, reperfusion injury, infectious a~ ;Lis, rheumatic fever and the like, and may - 30 reduce hemorrhage in acute promyelocytic leukemia and the like.
For each of the uses, the compounds of Formula (I) and (F)-actin shortening ~ro~ s, may be ~tlmini.stered orally, topically, parenterally, by inh~l~tion spray or rectally in dosage unit Formulations cont~ining conventional non-toxic ph~rm~ceutically acceptable carriers, adjuvants and vehicles. The term parenteral as used herein includes 2 ~ 84 385 subcutaneous injections, intravenous, intramuscular, intrasternal injection or infusion techniques. In addition to the treatment of warm-blooded ~nim~l~ such as mice, rats, horses, dogs, cats, etc., the compounds of the invention are effective in the treatment of hllm~n~.
The ph~ ceutical compositions cont~ining the active ingredient may be in a form suitable for oral use, for example, as tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or elixirs. Compositions intended for oral use may be prepared according to any method known to the art for the manufacture of pharmaceutical compositions and such compositions may contain one or more agents selected from the group consisting of sweetening agents, flavoring agents, coloring agents and preserving agents in order to provide ph~rm~ceutically elegant and palatable preparation. Tablets contain the active ingredient in admixture with non-toxic ph~ ceutically acceptable excipients which are suitable for the manufacture of tablets.
These excipients may be, for example, inert diluents, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; gran~ ting and disintegrating agents, for example, corn starch, or alginic acid; binding agents, for example starch, gelatin or acacia, and lubricating agents, for example, magnesium stearate, stearic acid or talc. The tablets may be uncoated or they may be coated by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, a time delay material such as glyceryl monostearate or glyceryl distearate may be employed.
Form~ tions for oral use may also be presented as hard gelatin capsules wherein the act*e ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules wherein the active ingredient is mixed with water or an oil medium, for example, peanut oil, liquid paraffin, or olive oil.
Aqueous suspensions contain the active materials in admixture with excipients suitable for the m~mlf~cture of aqueous WO 95/24207 PCTtUS95/02938 suspensions. Such excipients are suspending agents, for example, sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethyl-cellulose, sodium ~lgin~te, polyvinylpyrrolidone, gum tragacanth and gum acacia; dispersing or wetting agents may be a naturally-occurring phosphatide, for example, lecithin, or condensation products of an alkylene oxide with fatty acids, for example, polyoxyethylene stearate, or condensation products of ethylene oxide with long chain aliphatic alcohols, for example, heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, for example, polyoxyethylene sorbitan monooleate. The said aqueous suspensions may also contain one or more preservatives, for example, ethyl, or n-propyl, p-hydroxy-benzoate, one or more coloring agents, one or more flavoring agents, and one or more sweetening agents, such as sucrose or saccharin.
Oily suspension may be for T ~ ted by suspending the active ingredient in a vegetable oil, for example, arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin.
The oily suspensions may contain a thickening agent, for example, beeswax, hard paraffin or cetyl alcohol. Sweetening agents such as those set forth above, and flavoring agents may be added to provide a palatable oral l~r~aration. These compositions may be preserved by the addition of an antioxidant such as ascorbic acid.
Dispersible powders and granules suitable for preparation of an aqueous suspension by the addition of water provide the active ingredient in ~lmixtllre with a dispersing or wetting agent, suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are exemplified by those already mentioned above. Additional excipients, for example, sweetening, flavoring and coloring agents, may also be present.
The ph~rm~ceutical compositions of the invention may also be in the form of oil-in-water emulsions. The oily phase may be a vegetable oil, for example, olive oil or arachis oils, or a mineral oil, for 2l 84385 --example, liquid paraffin or mixtures of these. Suitable emulsifying agents may be naturally-occurring gums, for example, gum acacia or gum tragacanth, naturally-occurring phosphatides, for example, soy bean, lecithin, and esters or partial esters derived from fatty acids and hexitol anhydrides, for example, sorbitan monooleate, and condensation products of the said partial esters with ethylene oxide, for example, polyoxyethylene soll,i~l monooleate. The emulsions may also contain sweetening and flavoring agents.
Syrups and elixirs may be formulated with sweetening agents, for example, glycerol, propylene glycol, sorbitol or sucrose.
Such formulations may also contain a demulcent, a preservative and flavoring and coloring agents. The ph~rm~ceutical compositions may be in the form of a sterile injectable aqueous or oleagenous suspension.
This suspension may be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents which have been mentioned above. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally-acceptable diluent or solvent, for example, as a solution in 1,3-butane diol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution glucose in water and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil may be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables.
The ph~rm~ceutical compositions of this invention may also be ~tlmini~tered in the form of suppositories for rectal ~lmini.ctration of the drug. These compositions can be prepared by mixin~ the drug with a suitable non-irritating excipient which is solid at ordhlaly temperatures but liquid at the rectal temperature and will therefore melt in the rectum to release the drug. Such materials are cocoa butter and polyethylene glycols.
For topical use, creams, ointments, jellies, solutions or suspensions, etc., cont~inin~ the anti-infl~mm~tory agents are employed.

WO 95/24207 PCTtUS95/02938 - 21 8~385 The amount of active ingredient(s) that may be combined with the carrier materials to produce a single dosage form will vary depending upon the host treated and the particular mode of ~1mini~tration. For example, a formulation intended for the oral ~lmini~tration of hllm~n~ may contain from S mg to 2000 mg or 5000 mg of each active agent(s) compounded with an appropriate and convenient amount of carrier material which may vary from about 5 to about 95 percent of the total composition. For purposes of this o specification, this broad dosage range is specif1cally intended to include, but is not limited to, range of 5 mg to 2000 mg; 25 mg to 2000 mg; 5 mg to 1000 mg; 25 mg to 1000 mg; 5 mg to 500 mg; and 25 mg to 500 mg. Dosage unit forms will generally contain between from about 25 mg to about 500 mg of active ingredient(s).
Furthelll.ore, it is also possible that most effective treatment may warrant ~lmini~tration of an initial dosage of one range (e.g., 1-5 mg of active agent per kg of patient weight) followed by ~tlmini~tration of a second range (e.g., 0.1 to 1 mg of active agent per kg of patient weight).
It will be understood, however, that the specific dose level for any particular patient will depend upon a variety of factors including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of ~lmini~tration, route of ~lmini~tration, rate of excretion, drug combination and the severity of 25 the particular disease undergoing therapy.
With specific regard to (F)-actin shortening proteins, in particular gelsolin, the dosage can be calculated in the following manner. The normal blood gelsolin concentration is 2.4 ,uM (2.4 ~moVL), and the normal blood DBP concentration is 5 ,uM (5 ,umol/L).
30 Thus, the total blood actin-binding capacity (ABC) is approxim~tely 7.5 ~lmoVL. The blood volume if 6% of the body weight, hence a 70 Kg person has 4.2 liters of blood and thus (4.2 L x 7.5 ~mol/L) 31.5 ,umols ABC. Since DBP and gelsolin are distributed throughout the extracellular space (which is 10% of the body weight, the body contains (7.5 x 7) 52.5 ,umols ABC.

2 1 843~5 - so -It may be desired to ~lmini.~ter between 32 and 53 llmols of an actin binding (herein actin shortening) compound (or 0.46 ~mol/kg body weight) to cover total complexing or depletion of endogenous ABC. Since 0.425 mg of actin is equal to 1 ,umol, and since there is 4.86 mg actin per gram of skeletal muscle, each gram of muscle contains 11.3 ,umol actin or 4.6 grams of muscle destruction could neutralize total body ABC. However, because the toxic effects of actin are presumably local (e.g., inhibition of clot lysis), sequestered or kinetically determined (e.g., actin permeates an organ faster than binding proteins neutralize it), it is likely that a theoretically minimum dose will have to be adjusted upward in order to achieve kinetically favorable therapeutic effects. The kinetic effect can be important, for example, since hemolysis of about half of erythron, which should liberate only 4.2 ,umol of actin, reduces the pl~mi~ gelsolin concentration by half acutely (Smith, et al., Blood 72:214-2181 (1988)), suggesting slow equilibration between extravascular and blood compartments. Conversely, a therapeutically effective state, capable of breaking up local deposits of actin, may be achievable only by a transient pulse of a high concentration of actin-binding molecules.
The compounds of the invention can be ~(lmini~tered in any ayyroyliate ph~ cological carrier for a(lmini~tration. They can be ~lmini~tered in any form that effects prophylactic, palliative, preventative or curing conditions of tissue injury in h~lm~n~ and ~mm~l~
The following example illustrates the preparation of the compounds of Formula I useful in the method of treatment of the present invention, but does not limit the scope of the invention. Starting materials may be optionally prepared as disclosed in EPO 337 549 published.
October 18, 1989 which is hereby incorporated by reference. Where ayyloyliate, compounds may be produced and used in the form of ph~rm~ceutically acceptable salts. For example, the basic compounds may be used in the form of a hydrochloride or mesylate or other acceptable salt. See Preformulation in Remington's Pharmaceutical Sciences, Mack Publi~hin~, Easton PA.

F

~o R1 ~\~ R2 NH ~,~

o N~\Q-C-CI

~ R2 NH~

- M

H-Y-N~

~N O~ \O-C-Y-N~R

,~R2 NH ~,~

WO 95/24207 PCI~/US95/02938 R ~ ~Q IOI OH

/ ~ \

oxalyl Br2CH2c0 chloride base Ex 3a Ex 1 A

Ex 3B HO-CH-CO2Bnb-C-O-CH2-C02-TFA Ex 1B
anisole \Q-C-O-CH-CO2Bn ,~
7 `Q-C-O-CH2-cO2H
, R7 Ex 4 H2/cat R8 ~ CDI
~1l 1l ,R7 -C-O-CH2-C-N~ R

- SCHEME 2 CONrr'D

s 1) oxalyl chloride ,R7 2) HN\ Ex 5 s5~ 1l ICH3 1l R7 -C-O-CH-C-N

The glycolic acid derivatives described herein can be prepared according to the following scheme. The starting acid (as 2s carboxylate anion) may be alkylated (Ex lA) with a suitably protected a-halo acetic acid derivative to give the glycolate ester _ which can be deprotected (Ex lB) to the glycolic acid ester 5. Treatment of 5 with an amine lltili7.in~ a condensing agent such as dicyclohexylcarbodiimide or carbomyldiinidazole (Ex 2) affords the deserved amide 6. Alternately, 30 the starting acid 1 may be converted to its acid chloride 2 (Ex 3A) and - treated with a suitably protected a-hydroxyalkanoic acid (Ex 3B) in the presence of base to give the protected ester 7. Deprotection (Ex 4), followed by conversion to the acid chloride and treatment with the ayyroyliate amine (Ex 5) affords the desired amide 9.

21 ~4385 A. t-Butoxycarbonylmethyl [S-(R*, S*)]4-((3,3-diethyl-1-(((1-(4-methylphenyl)butyl)amino)-carbonyl)-4-oxo-2-azetidinyl)-oxy)benzoate To a solution of 0.806 gm of [S-(R*, S*)] 4-((3,3-diethyl-1-((( 1 -(4-methylphenyl)butyl)amino)-carbonyl)-4-oxo-2-azetidinyl)oxy)-benzoic acid in ~3ml DMF is added 0.23 gm. triethylamine followed by 0.50 gm of t-butyl bromoacetate and the mixture stirred overnight at room temperature. Ethyl acetate (25 ml) is then added and the resultant mixture is washed with 2 x 10 ml water, 10 ml saturated sodium bicarbonate, and 20 ml brine. The organic layer is dried through sodium sulfate and concentrated in vacuo. Chromatography on Silica gel 60 (350 ml column) and elution with 10% ethyl acetate in hexanes gave 0.67 gm of the t-Butoxycarbonylmethyl [S-(R*, S*)]4-((3,3-diethyl- 1-(((1 -(4-methylphenyl)butyl)amino)carbonyl)-4-oxo-2-azetidinyl)oxy)benzoate .
In a similar m~nner can be prepared 2-(dimethylamino)-2-oxoethyl, (S-(R* ,S*))-4-((3 ,3 -Diethyl- 1-(((1 -(4-methylphenyl)butyl)-amino)carbonyl)-4-oxo-2-azetidinyl)oxy)benzoate (Compound 6), and 2-(N-methylacetamido)ethyl. ~2-(R*,S*)}- 4-{ {3,3-Diethyl-1-{ { { 1-(4-methylphenyl)butyl } amino ) carbonyl } -4-oxo-2-azetidinyl } oxy } -benzoate, (Compound 7).

B. Carboxymethyl [S-(R*,S*)]4-((3,3-diethyl-1- (((1-(4-methyl-phenyl)butyl)amino)carbonyl)-4-oxo-2-azetidinyl)-oxy)benzoate To the above ester is added 2 ml of anisole and the resulting mixture is cooled in an ice bath and 5 ml of ice cold trifluoroacetic acid is added. The reaction mixture is stirred cold for three hours then allowed to come to room temperature. After 30 minlltes, the reaction mixture is concentrated in vacuo and the residue chromatographed on silica gel 60. Elution with 20% ethyl acetate in hexanes cont~ining 1% acetic acid gives 0.53 gm of desired carboxymethyl [S-(R* ,S *)]4-((3,3 -diethyl- 1 -(((1 -(4-methyl-phenyl)butyl)amino)carbonyl)-4-oxo-2-azetidinyl)oxy)benzoate.

Compound 2 Analysis: C28H34N2o7+o.3H2o Calc: C,65.19;H,6.75;N,5.43 Found: C, 65.30; H, 6.76; N, 5.23.

Compound 6 o Analysis: C30H39N306+0.5H20 Calc: C,65.91;H,7.38;N,7.60 Found: C, 65.71; H, 7.63; N, 7.50.

Compound 7 Analysis: C31H41N306+0.6EtOAc Calc: C, 66.35; H, 7.64; N, 6.95 Found: C, 66.52; H, 7.89; N, 6.83.

2-(bis(2-hydroxyethyl)amino)-2-oxoethyl(S-(R*,S*))-4-((3,3-diethyl- 1 -(((1 -(4-methyl-phenyl)butyl)amino)-carbonyl)-4-oxo-2-azetidinyl)oxy) benzoate To a solution of 0.125 gm of the acid from lB in 2-3 ml of methylene chloride is added 0.050 gm of carbonyldiimidazole. The mixt~lre is stirred for 30 minutes at room temperature at which time 0.060 gm of diethanol~mine is added along with 1 ml of DMF and 2 ml of methylene chloride. The resulting mixture is stirred overnight at room temperature then concentrated in vacuo. Silica gel chromatography of the residue using 2.5 to 5.0% methanol in methylene chloride gives 0.123 gm of the desired Compound 3, 2-(bis(2-hydroxy-ethyl)amino)-2-oxoethyl(S-(R*,S*))-4-((3,3-diethyl-1-(((1-(4-methyl-phenyl)butyl)amino)carbonyl)-4-oxo-2-azetidinyl)oxy)benzoate .

21 843~5 Compound 3 Analysis: C32H43N3O8~ +0.4H20 Calc: C, 63.53; H, 7.30; N, 6.95 Found: C, 63,51; H, 7.45; N, 6.95.

Similarly were prepared Compound 4 Analysis: C31H40N4O7 Calc: C, 64.12; H, 6.94; N, 9.65 Found: C, 64.12; H, 7.18; N, 9.44.

Compound 5 Analysis: C32H43N309 +0.3H20 Calc: C, 62.08; H, 7.09; N, 6.79 Found: C, 61.89; H, 7.39; N, 6.88.

Compound 8 Analysis: C40H47N3o8 Calc: C, 68.85; H, 6.79; N, 6.02 Found: C, 68.79; H, 7.06, N, 5.88.

Preparation of 1 -Methyl-2-oxo-2-(phenylmethoxy)ethyl(2S-( l (S *) ,R* ,-(R)))-4-((3,3-diethyl- 1 -(((1 -(4-methylphenyl)butyl)amino)carbonyl)-4-oxo-2-azetidinyl~oxy) benzoate. Compound 10 To a solution of 1.0 gm [S-(R*, S*)]4-((3,3-diethyl-1-(((1-(4-methylphenyl)butyl)amino)carbonyl)-4-oxo-2-azetidinyl)oxy)benzoic acid in 10 ml methylene chloride is added 2 ml of oxalyl chloride followed by a catalytic amount of DMF. The reaction is stirred 1 hour at room temperature then concentrated in vacuo to yield the acid chloride which is used as is in the next step.

A solution of the above acid chloride in 10 ml of methylene chloride is cooled in an ice bath and a solution of 1.25 gm benzyl L-lactate and 2.0 gm of triethyl~mine in 10 ml of methylene chloride is added. The mixture is stirred at room temperature overnight then concentrated in vacuo. Chromatography of the residue on silica gel using methylene chloride as the eluent yields 0.795 of the desired 1-Methyl-2-oxo-2-(phenylmethoxy)ethyl (2S-(l(S*),R*,(R)))-4-((3,3-diethyl- 1-(((1 -(4-methylphenyl)butyl)amino)carbonyl)-4-oxo-2-azetidinyl)oxy)benzoate, Compound 10.
Analysis: C36H42N207 Calc: C, 70.34; H, 6.89; N, 4.56 Found: C, 70.45; H, 7.05; N, 4.48.

Preparation of 1-carboxyethyl [S-(R*,S*)] 4-((3,3-diethyl-1-(((1-(4-methyl-phenyl)butyl)amino)carbonyl)-4-oxo-2-azetidin-yl)oxy)benzoate A mixture of 0.69 gm of the benzylester prepared in Example 3 and 0.2 gm 10% Pd/C in 10 ml of EtOAc is treated with hydrogen at 40 psi. When the reaction is complete the n~ ule is filtered and concentrated in vacuo to yield 0.56 gm of l-carboxyethyl [S-(R*,S*)]4-((3,3-diethyl-1 -(((1 -(4-methyl-phenyl)butyl)amino)-carbonyl)-4-oxo-2-azetidinyl)oxy)benzoate, Compound 11.
Analysis: C29H36N2o7 Calc: C, 66.40; H, 6.92; N, 5.34 Found: C, 66.66; H, 7.26; N, 5.05.

2-(diethylamino)- 1 -methyl-2-oxoethyl[S-(R* ,S*)] -4-((3 ,3-diethyl- 1-5 (((1-(4-methyl-phenyl)butyl)amino)-carbonyl)-4-oxo-2-azetidinyl)oxy) benzoate The acid (.250 gm) from Example 4 is treated with oxalyl chloride according to the procedure of Example 3A and the corresponding acid chloride is obtained. This material is dissolved in 5 o ml methylene chloride and 0.4 ml of diethylamine added. After 1 hour the reaction mixture is concentrated in vacuo and the residue taken in ethyl acetate and washed with saturated sodium bicarbonate solution.
The organic layer is dried through sodium sulfate, concentrated and the residue chromatographed on silica gel. Elution with 5% of ethyl acetate in methylene chloride gives Compound 12.
Analysis: C33H45N306 Calc: C, 68.37; H, 7.82, N, 7.25 Found: C, 68.40; H, 7.93, N, 7.40.

(S(R*,S*))-l -(((4-((3,3-diethyl-1-(((1 -(4-methyl-phenyl)butyl)amino)-carbonyl)-4-oxo-2-azetidinyl)oxy)benzoyl)oxy)acetyl) L-proline 2s ---When benzyl L-lactate is replaced by L-proline benzyl ester hydrochloride and triethyl~mine in the procedure of Example 3 the corresponding amide with L-proline benzyl ester, Compound 8, is obtained.
Analysis: C40H47N308 Calc: C, 68.85; H, 6.79, N, 6.02 Found: C, 68.79; H, 7.06, N, 5.88.

WO 95/24207 2 1 8 4 3 8 5 PCT/US~5 ~2g38 Reduction of the material obtained in Fx~mple 6A
according to the procedure of Example 4 affords Compound 9.
Analysis: C33H41N308+0.5H20 Calc: C, 64.27; H, 6.86; N, 6.81 Found: C, 64.49; H, 6.90; N, 6.68.

[S-(R*,S*)] 1-(((4-((3,3-diethyl-1-(((1-(4-methyl- phenyl)butyl)amino) carbonyl-4 -oxo-2-azetidinyl)oxy)benzoyl)oxy)acetyl-N -benzyl-L-prolinamide Treatment of the acid obtained in Example 6B, Compound-9~ with oxalyl chloride according to Example 3A gives the corresponding acid chloride which when treated with benzylamine gives the desired benzyl amide, Compound 19.
Analysis: C40H4gN4O7 Calc: C, 68.95; H, 6.94; N, 8.04 Found: C, 68.93, H, 7.02; N, 7.96.

To a solution of the acid chloride (~clJarcd from 0.55 gm of [S-(R*, S*)] 4-((3,3-diethyl-1-(((1-(4-methylphenyl)butyl)amino)-carbonyl)-4-oxo-2-azetidinyl)oxy)benzoic acid according to the procedure of Example 3A) in 3 ml of methylene chloride is added 0.15 gm of N,N-dimethylaminoethanol. The reaction mixture is stirred overnight at room temperature, concentrated in vacuo, then taken up in ethyl acetate (25 ml) and washed with saturated sodium bicarbonate solution. The organic layer is dried through sodium sulfate and concentrated in vacuo. Silica gel chromatography of the residue using 2.5% methanol in methylene chloride g*es 0.59 gm of Compound 1, 2-(dimethylamino)ethyl (S-(R*,S*))-4-((3,3-diethyl-1-(((1-(4-methyl-phenyl)butyl)amino)carbonyl)-4-oxo-2-azetidinyl)oxy)benzoate WO 95t24207 PCT/US95/02938 Analysis: C30H41N3O5 Calc: C, 68.81; H, 7.89; N, 8.02 Found: C, 68.85; H, 8.09; N, 7.97.

When N,N-dimethylaminoethanol~mine is replaced by the appropriate amino alcohols the corresponding esters are obtained.

Compound 13 1-Dimethylamino-2-propyl [S-(R*,S*)]-4-[[3,3-diethyl-1 -[[[1 -(4-methyl-phenyl)butyl]amino]carbonyl]-4-oxo-2-azetidin-yl]oxy]benzoate Analysis: C31H43N305 Calc: C, 69.25; H, 8.06; N, 7.82 Found: C, 68.97; H, 8.01; N, 7.80.

Compound 14 3-Dimethylamino-l-propyl [S-(R*,S*)]-4-[[3,3-diethyl-1 -[[[1 -(4-methyl-phenyl)butyl]amino]carbonyl]-4-oxo-2-azetidin-yl]oxy]benzoate Analysis: C31H43N3O5 Calc: C, 69.25; H, 8.06; N, 7.81 Found: C, 68.85; H, 8.19; N, 7.72.

Compound 16 2-Diethylaminoethyl [S-(R*,S*)]-4-[[3,3-diethyl- 1 -[[[1 -(4-methyl-phenyl)butyl]amino]carbonyl]-4-oxo-2-azetidinyl] -oxy]benzoate Analysis: C32H45N306 Calc: C, 69.66; H, 8.22; N, 7.62 Found: C, 69.37; H, 8.41; N, 7.51.

Compound 17 2-(1-[4-morpholino]e~yl) [S-(R*,S*)]-4-[[3,3-diethyl-1-[[[1 -(4-methyl-phenyl)butyl]amino]carbonyl]-4-oxo-2-azetidinyl]oxy]benzoate WO 95/24207 PCTlU~5~ '93fi - 21 a438s Analysis: C32H43N306 Calc: C, 67.94; H, 7.66; N, 7.43 Found: C, 67.67; H, 7.90; N, 7.26.

Compound 18 4-dimethylaminobutyl [S-(R*,S*)]4-[[3,3-diethyl-1-[[[1-(4-methyl-phenyl)butyl]amino]carbonyl]4-oxo-2-azetidinyl]oxy]benzoate Analysis: C32H45N305 +0.2 H20 Calc: C, 69.21; H, 8.24; N, 7.56 Found: C. 69.35; H, 8.24; N, 7.29.

Compound 20 2-dimethylamino-2-methyl-1-propyl [S-(R*,S*)]-4-[[3,3-diethyl-1 -[[[1 -(4-methyl-phenyl)butyl]amino]carbonyl]-4-oxo-2-azetidinyl]oxy]benzoate Analysis: C32H45N3O5 Calc: C, 69.66; H, 8.22; N, 7.62 Found: C, 69.52; H, 8.47; N, 7.59.

Compound 21 2-(diisopropylamino)ethyl [S-(R*,S*)]-4-[[3,3-diethyl-1-[[[1 -(4-methyl-phenyl)butyl]amino]carbonyl]4-oxo-2-azetidinyl]oxy]benzoate Analysis: C34H49N305 Calc: C, 70.44; H, 8.52; N, 7.25 Found: C, 70.28; H, 8.76; N, 7.13.

Compound 22 Benzyl [S-(R*,S*)]4-[2-[[4-[[3,3-diethyl-1-[[[1-(4-methylphenyl)butyl]amino]carbonyl]4-oxo-2-azetidinyl]oxy]-benzoyl]oxy]-ethyl]-l -Piperazine-carboxylate Analysis: C40HsoN407 Calc: C, 68.75; H, 7.21; N, 8.02 Found: C, 68.39; H, 7.30; N, 7.84.

Compound 23 2-(dibutylamino)ethyl [S-(R*,S*)]-4-[[3,3-diethyl-1-[[[1-(4-methyl-phenyl)-butyl]amino]carbonyl]-4-oxo-2-azetidin-yl]oxy]benzoate Analysis: C36Hs3N305 Calc: C, 71.14; H, 8.79; N, 6.91 Found: C, 71.00; H, 9.03; N, 6.81.

Compound 24 [S-(R*,S*)]-6-(dimethylamino)hexyl-4-[[3,3-diethyl-1-[[[1 -(4-methyl-phenyl)butyl]amino]carbonyl] -4-oxo-2-azetidin-yl]oxy]-benzoate Analysis: C34H49N305+1H20 Calc: C, 68.31; H, 8.60; N, 7.03 Found: C, 68.34; H, 8.29; N, 6.86.

Compound 26 2-(4-me~yl-1-piperazinyl)ethyl[S-(R*,S*)]-4-[[3,3-diethyl-1-[[[1-(4-methyl-phenyl)butyl]amino]carbonyl]-4-oxo-2-azetidinyl]oxy]benzoate, Analysis: C33H46N405+0.8H20 Calc: C, 66.82; H, 8.09; N, 9.44 Found: C, 67.28; H, 8.10; N, 8.96.

Compound 28 2- (diphenylamino)ethyl [S -(R * ,S *)] -4- [ [3,3 -diethyl- 1 -[[[1 -(4-methylphenyl)-butyl]amino]carbonyl] -4-oxo-2-azetidin-yl]oxy]benzoate Analysis: C40H4sN30s +1.4H20 Calc: C, 71.40; H, 7.16; N, 6.25 Found: C, 71.62; H, 6.99; N, 5.99.

Compound 29 2-(di-2-propenylamino)ethyl [S-(R*,S*)]-4-[~3,3-diethyl-1 -[[[1 -(4-methylphenyl)-butyl]amino]carbonyl]-4-oxo-2-azetidin-yl]oxy]benzoate, Analysis: C34H45N305 Calc: C, 70.93; H, 7.88; N, 7.30 Found: C, 71.18; H, 8.06; N, 7.34.

Compound 30 2-(dimethylamino)-2-phenylethyl [S-(R*,-S*)]-4-[[3,3-diethyl-l-[[[l- (4-methyl- phenyl)butyl]amino]carbonyl]-4-oxo-2-azetidinyl]oxy]benzoate Analysis: C36H45N3O5 Calc. C, 72.09; H, 7.56; N, 7.00.
Found: C, 71.75; H, 7.67; N, 6.70.

Compound 31 2-[methyl(phenylmethyl)amino]ethyl [S-(R*,S*)]-4-o [[3,3-diethyl-1-[[[1-(4-methyl-phenyl)butyl]-amino]carbonyl]-4-oxo-2-azetidinyl]oxy]benzoate When [S-(R*,S*)]-4-[[3,3-diethyl-1-[[[1-(4-methyl-phenyl) butyl]amino]carbonyl]-4-oxo-2-azetidinyl]oxy]-2,6-dimethyl benzoic acid is used in place of [S(R*,S*)]-4-[[3,3-diethyl-1-[[[1-(4-methyl-phenyl)butyl]amino]carbonyl]-4-oxo-2-azetidinyl]oxy]benzoic acid in the procedure of Example 3A and allowed to react with the ayyroyliate amino alcohols the following esters are obtained.

Compound 304 2-(dimethylamino)ethyl [S,(R*,S*)]-4-[[3,3-diethyl-1 -[[[1 -(4-methylphenyl)-butyl]amino]carbonyl]-4-oxo-2-azetidin-yl]oxy]-2,6-dimethyl-benzoate Compound 305 2-~diethylamino)ethyl [S-(R*,S*)]-4-[[3,3-diethyl-1 -2 5 [ [ [1 -(4-methylphenyl)-butyl] amino] carbonyl] -4-oxo-2-azetidin-yl]oxy]-2,6-dimethyl-benzoate Compound 306 2-[bis(l-methylethyl)amino]-ethyl [S-(R*,S*)]-4-[[3,3-diethyl-1 -[[[1 -(4-methylphenyl)butyl]amino]-carbonyl] -4-oxo-2-azetidinyl]oxy] -2,6-dimethyl-benzoate Tre~trnen~ of the acid [S-(R*,S*)]-4-[[3,3-diethyl-1-[[[1-(4-methylphenyl)butyl]amino]carbonyl]-4-oxo-2-azetidinyl]oxy]-benzene-acetic acid with oxalyl chloride according to the procedure of Example 3A affords the corresponding acid chloride which when allowed to react with the a~rol~liate amino alcohol according to the procedure of Example 8 gives the following amino esters:

Compound 32 Analysis: C31H43N3O5 Calc: C, 69.25; H, 8.06; N, 7.82 Found: C, 69.02; H, 7.86; N, 7.74.

Compound 33 Analysis: C32H45N3O5 Calc: C, 69.66; H, 8.22; N, 7.62 Found: C, 69.10; H, 8.17; N, 7.50.

Compound 34 Analysis: C33H47N305 Calc: C, 70.06; H, 8.38; N, 7.43 Found: C, 69.70; H, 8.41; N, 7.05.

Compound 35 Analysis: C33H45N306 Calc: C, 68.37; H, 7.82; N, 7.25 Found: C, 68.55; H, 8.19; N, 7.08.

Compound 36 Analysis: C32H45N305 Calc: C, 69.66; H, 8.22; N, 7.62 Found: C, 69.60; H, 8.49; N, 7.55.

To a solution of 0.247 g of Compound 1 in 2 ml of ethyl acetate is added 0.125 gm of m-chloroperoxy benzoic acid. After 30 es at room temperature the reaction mix~lre is concentrated in vacuo. Chromatography of the residue on silica gel using methylene chloride/methanol/water 85/15/1.5 gives the desired N-oxide Compound 15.
Analysis: C3oH4lN3o8+l.4H2o Calc: 63.79; H, 7.82; N, 7.44 Found: 63.89; H, 7.85; N, 7.27.

[S-(R*,S*)] 2-[4-[[[2-(dimethylamino)-ethyl]amino]-carbonyl]phenoxy]-3,3 -diethyl-N- [1 -(4-methylphenyl)-butyl] -4-oxo- 1 -azetidinecarbox -amide To a solution of 0.104 g carbonyldiimidazole in 2 ml methylene chloride is added a solution of 0.227 g of [S-(R*,S*)]~-((3,3-diethyl-1-((-1-(4-methylphenyl)butylamino)carbonyl)-4-oxo-2-azetidinyl)-oxy)benzoic acid in 3 ml methylene chloride. The mixture is stirred at ambient tempe.a~ule for 30 mim-tes at which time 0.100 g of N,N-dimethyl-ethylene~ mine is added. After stirring overnight at room temperature the reaction mixture is poured into benzene (50 ml) and washed with water. The organic layer is separated, dried through sodium sulfate and concentrated in vacuo. Silica gel chromatography using 5% methanol in methylene chloride yields 0.160 g of 2-[4-[[[2-(dimethylamino)ethyl]amino]-carbonyl]phenoxy]-3,3-diethyl-N-[1-(4-methylphenyl)butyl]-4-oxo-1-azetidinecarbox~mitle.
(Compound 73).
Analysis: C30H42N404+0.4H20 Calc: C, 68.00; H, 8.14; N, 10.57 Found: C, 68.01; H, 8.18; N, 10.62.

WO 95/24207 2 1 8 4 3 8 5 PCTtUS95/02938 A. (S-(R*,S*))-4-((3,3-diethyl-1-((1-(4-methylphenyl)butyl-amino)carbonyl)-4-oxo-2-azetidinyl)-oxy)-benzoyl chloride To a solution of 0.150 g of [S-(R*,S*)] 4-((3,3-diethyl-1-(((1 -(4-methylphenyl)butyl)amino)-carbonyl)-4-oxo-2-azetidinyl)-oxy)benzoic acid in 5 ml methylene chloride cont~ining a catalytic o amount of dimethylformamide is added 0.5 ml of oxalyl chloride.
The mixture is stirred at room temperature for 30 minutes and then concentrated in vacuo to yield (S-(R*,S*))-4-((3,3-diethyl-1-((1-(4-methylphenyl)butylamino)carbonyl)-4-oxo-2-azetidinyl)oxy)-benzoyl chloride.

B [S-(R*,S*)]-2-[4-[[[2-(dimethylamino)ethyl]methyl-amino]carbonyl]-phenoxy]-3,3-diethyl-N-[1-(4-methylphenyllbutyll-4-oxo- 1 -azetidinecarboxamide The above acid chloride is dissolved in 3 ml methylene 20 chloride and a solution of 0.20 gm of N,N,N'-trimethylethylene-min~ in 2 ml methylene chloride is added. The mixture is stirred overnight and then concentrated in vacuo. The residue is extracted between ethyl acetate and saturated sodium bicarbonate solution.
The organic layer is dried through sodium sulfate and concentrated 25 in vacuo. Silica gel chromatography of the residue (5% methanol in methylene chloride) affords 0.117 g of [S-(R*,S*)]-2-[4-[[[2-(dimethylamino)ethyl]methylamino]carbonyl]phenoxy]-3,3-diethyl-N-[1-(4-methylphenyl]butyl]-4-oxo-1-azetidinecarboxamide.
(Compound 75).
Analysis: C3 1 H44N404+0.5H20 Calc: C, 68.23; H, 8.31; N, 10.27 Found: C, 68.20; H, 8.41; N, 10.10.

When N,N,N'-trimethylethylene~ min~ of Example 1 lb is replaced by the a~prol~-iate amines, there is obtained the corresponding amides.

1) Compound 76 Analysis: C32H46N4O4 Calc: C, 69.79; H, 8.42; N, 10.17 Found: C, 69.02; H, 8.60; N, 9.54.

2) Compound 77 Analysis: C32H46N404 +0.75 H20 Calc: C, 68.12; H, 8.49; N, 9.93 Found: C, 68.22; H, 8.48; N, 10.17.

3) Compound 78 Analysis: C32H44N405 Calc. C, 68.06; H, 7.85; N, 9.92 Found: C, 67.84; H, 8,07; N, 9.62.

4) Compound 79 Analysis: C33H46N4O5 +H2O
Calc: C, 66.42; H, 8.11; N, 9.39 Found: C, 66.73; H, 8.19; N, 9.23.

S) Compound 80 Analysis: C35H52N406 Calc: C, 67.28; H, 8.39; N, 8.97 Found: C, 67.08; H, 8.77; N, 8.41.

6) Compound 81 Analysis: C33H48N404 +H2O
Calc: C, 68.01; H, 8.65; N, 9.61 Found: C, 68.42; H, 8.59; N, 9.17.

21 843~5 7) Compound 82 Analysis: C36H46N4O4 +0.5H20 Calc: C, 71.14; H, 7.79; N, 9.21 Found: C, 71.41; H, 7.68; N, 9.10.

8) Compound 83 Analysis: C32H46N404 +1.2H20 Calc: C, 67.15; H, 8.52; N, 9.79 Found: C, 67.21; H, 8.26; N, 9.47.

9) Compound 84 15 10) Compound 86 Analysis: C35H52N4O4 Calc: C, 70.91; H, 8.84; N, 9.45 Found: C, 70.37; H, 8.84; N, 8.77.

20 11) Compound 89 Analysis: C31H39N504 +0.7H20 Calc: C, 66.71; H, 7.29; N, 12.54 Found: C, 66.91; H, 7.40; N, 12.14 12) Compound 90 Analysis: C33H46N404 +0.8H20 Calc: C, 68.67; H, 8.31; N, 9.70 Found: C, 68.82; H, 8.11; N, 9.70.

30 13) Compound 91 Analysis: C34H48N404 + 0.3H20 Calc: C, 70.11; H, 8.41; N, 9.61 Found: C, 70.17; H, 8.64; N, 9.33.

-14) Compound 92 Analysis: C30H42N404 + 1.2H20 C alc: C, 66.14; H, 8.22; N, 10.29 Found: C, 66.18; H, 8.12; N, 10.31.

15) Compound 93 Analysis: C32H44N406 + 0.3H20 C alc: C, 65.57; H, 7.67; N, 9.56 Found: C, 65.72; H, 7.50; N, 9.34.

16) Compound 94 Analysis: C32H44N4O4 + 0.3H20 Calc: C, 70.04; H, 8.08; N, 10.21 Found: C, 70.34; H, 8.90; N, 8.93.

17) Compound 9S
alysis: C 33 H 46 N 4 0 4 + .S H 20 Calc: C, 69.32; H, 8.28; N, 9.80 Found: C, 69.41; H, 8.25; N, 9.58.

18) Compound 99 Analysis: C3gH54N404 + l.SH20 C alc: C, 69.37; H, 8.72; N, 8.51 Found: C, 69.48; H, 8.44; N, 8.36.

19) Compound 102 alysis: C 40 H 49 N 5 0 6 + 1.0 H 2 0 C alc: C, 67.30; H, 7.20; N, 9.81 Found: C, 67.50; H, 7.24; N, 9.53.

20) Compound 104 alysis: C 3 2 H 41 N 5 0 4 + 0.75 H 2 0 C alc: C, 67.04; H, 7.47; N, 12.21 Found: C, 67.16; H, 7.56; N,ll.9S.

21) Compound 105 ~41lalysis: C 32 H 44 N 4 0 5 + 0.5 H 2 0 C alc: C, 66.99; H, 7.91; N, 9.77 Found: C, 67,00; H, 8.25; N, 9.50.

22) Compound 106 A nalysis: C 3 2 H 45 N 5 0 5 +0.8H20 C alc: C, 64.69; H, 7.90; N, 11.78 Found: C, 64.93; H, 8.25; N, 11.12.

23) Compound 107 Analysis: C31H44N306S +0.3H20 Calc: C, 61.42; H, 7.41; N, 9.24 Found: C, 61.43; H, 7.54; N, 9.05.

24) Compound 110 Analysis: C35H44N404 C alc: C, 71.89; H, 7.58; N, 9.58 Found: C, 71.65; H, 7.55; N, 9.34.

25) Compound 111 alysis: C 34 H 42 N 4 0 4 + 0.5 H 2 0 C alc: C, 70.44; H, 7.47, N, 9.66 Found: C, 70.82, H, 7.46; N, 9.20.

26) Compound 113 ~41lalysis: C 34 H 42 N 4 0 4 + 0.3 H 2 0 Calc: C, 70.88; H, 7.45; N, 9.72 Found: C, 71.12; H, 7.44; N, 9.32.

27) Compound 115 A nalysis: C34H42N404 +0.7H20 C alc: C, 69.99; H, 7.50, N, 9.60 Found: C, 70.14, H, 7.63; N, 9.25.

WO 95124207 PCT/U$95/02938 28) Compound 116 Analysis: C34H46N404 +1.4H20 Calc: C, 68.06; H, 8.19; N, 9.33 Found: C, 68.40; H, 8.14; N, 8.87.

29) Compound 117 Analysis: C40H51N5O6 +0.6H20 Calc: C, 67.79; H, 7.42; N, 9.88 Found: C, 67.81; H, 7.58; N, 9.76.

30) Compound 118 Analysis: C33H47N504 +0.7H20 Calc: C, 67.14; H, 8.26; N, 11.86 Found: C, 67.54; H, 8.51; N, 11.28.

31)Compound119 Analysis: C35H48N404 +1.25H20 Calc: C, 68.76; H, 8.32; N, 9.16 Found: C, 69.07; H, 8.19; N, 8.75.

32) Compound 121 Analysis: C34H4gN4O4 +lH20 Calc: C, 68.66; H, 8.47; N, 9.42 Found: C, 69.02; H, 8.32; N, 9.06.
2s 33) Compound 125 Analysis: C37H48N404 +0.5H20 Calc: C, 71.47; H, 7.94; N, 9.01 Found: C, 71.65; H, 7.91; N, 8.73.

34) Compound 126 Analysis: C41H54N405 +2H20 Calc: C, 68.83; H, 8.25; N, 7.64 Found: C, 69.03; H, 7.79; N, 7.50.

35) Compound 132 alysis: C 35 H 50 N 4 0 4 C alc: C, 71.15; H, 8.53; N, 9.48 Found: C, 70.90; H, 8.74; N, 9.12.

36) Compound 133 alysis: C 4 0 H 52 N 4 0 4 + 0.9 H 2 0 C alc: C, 71.80; H, 8.10; N, 8.37 Found: C, 71.86; H, 8.17; N, 8.18.

37) Compound 137 A nalysis: C37H48N404 +0.8H20 C alc. C, 70.85; H, 7.97; H, 8.93 Found: C, 71.01; H, 7.97; N, 8.54.

38) Compound 139 A nalysis: C 39 H 51 N 5 0 4 +0.8H20 C alc: C, 70.09; H, 7.93; N, 10.48 Found: C, 70.18; H, 7.79; N, 10.42.

39) Compound 142 Analysis: C38H55N506 C alc: C, 67.33; H, 8.18; N, 10.33 Found: C, 67.02; H, 8.31; N, 9.89.

40) Compound 143 A nalysis: C34H51N504 C alc: C, 68.77; H, 8.66; N, 11.80 Found: C, 68.57; H, 8.50; N, 11.53.

41) Compound 144 ~nalysis: C 38 H 59 N 5 0 4 + 0.4 H 2 0 C alc: C, 69.45; H, 9.17; N, 10.65 Found: C, 69.69; H, 9.02; N, 10 35.

WO 95/24207 PCT/US9~/02938 42) Compound 145 ~41lalysis: C 36 H 47 N 5 0 4 + H 2 0 C alc: C, 68.44; H, 7.82; N, 11.08 Found: C, 68.69; H, 7.74; N, 10.76.

43) Compound 148 ~41lalysis: C 36 H 47 N 5 0 4 + 0.4 H 2 0 C alc: C, 69.62; H, 7.56; N, 11.27 Found: C, 69.79; H, 7.70; N, 11.12.

44) Compound 149 ~ alysis: C 39 H 50 N 4 0 4 + 0.4 H 2 0 C alc: C, 72.50; H, 7.93; N, 8.67 Found: C, 72.44; H, 7.99; N, 8.87.

45) Compound 150 ~41lalysis: C 36 H 47 N 5 0 4 + 0.3 H 2 0 C alc. C, 69.83; H, 7.74; N, 11.31 Found: C, 69.93; H, 7.65, N, 11.10.

46) Compound 151 A nalysis: C33H47N504 C alc: C, 68.60; H, 8.20; N, 12.12 2s Found: C, 68.40; H, 8.16; N, 11.90.

47) Compound 245 ~41lalysis: C 36 H 44 N 4 0 4 Calc: C, 72.46; H, 7.43; N, 9.39 Found: C, 72.49; H, 7.49; N, 9.25.

48) Compound 246 Analysis: C37H46N404 +0.25H20 C alc: C, 72.26; H, 7.61; N, 9.10 Found: C, 72.35; H, 7.83; N, 8.73.

49) Compound 154 Analysis: C35H4gN4O4 +0.8H20 Calc: C, 69.70; H, 8.29; N, 9.29 Found: C, 69.65; H, 8.27; N, 9.35.

50) Compound 158 Analysis: C35H48N404 +0.5H20 Calc: C, 73.29; H, 7.65; N, 8.33 Found: C, 73.71, H, 7.75, N, 7.75.

51) Compound 159 Analysis: C35H4gN4O4 Calc: C, 73.09; H, 7.66; N, 8.31 Found: C, 73.40; H, 7.75; N, 7.80.

52) Compound 160 Analysis: C38H48N404 +1.0H20 Calc: C, 70.78; H, 8.12; N, 8.68 Found: C, 71.00; H, 8.05; N, 8.59.

53) Compound 161 Analysis: C43H52N404 +lH20 Calc: C, 73.05; H, 7.70; N, 7.92 Found: C, 73.29; H, 7.95; N, 7.37.

54) Compound 166 Analysis: C33H46N404 +1.5H20 Calc. C, 67.20; H, 8.37; N, 9.50 Found: C, 67.38; H, 7.98; N, 9.41.

55) Compound 171 Analysis: C36H52N406 +1.6H20 Calc: C, 64.95; H, 8.36; N, 8.41 Found: C, 65.26; H, 8.15; N, 8.07.

56) Compound 177 Analysis: C3gH47N5O4 Calc: C, 71.56; H, 7.43; N, 10.98 Found: C, 71.64; H, 7.62; N, 10.93.

57) Compound 178 Analysis: C3gH50N4O4 Calc: C, 72.81; H, 8.04; N, 8.94 Found: C, 72.96; H, 8.17; N, 8.83.

58) Compound 179 Analysis: C3gH47N5O4 Calc: C, 71.56; H, 7.43; N, 10.98 Found: C, 72.00; H, 7.55; N, 10.87.

59) Compound 180 Analysis: C3gH47F3N4O4 Calc: C, 67.04; H, 6.96; N, 8.23 Found: C, 67.02; H, 7.25; N, 8.23.

60) Compound 181 Analysis: C3gH47F3N4O4 Calc: C, 67.04; H, 6.96; N, 8.23 Found: C, 66.63; H, 6.98; N, 7.94.

61)Compoundl82 Analysis: C37H47F N404 Calc: C, 70.45; H, 7.51; N, 8.88 Found: C, 70.28; H, 7.74; N, 8.82.

62) Compound 185 Analysis: C34H4gN4O4 Calc: C, 70.80; H, 8.39; N, 9.71 Found: C, 70.44; H, 8.45; N, 9.51.

63) Compound 186 Analysis: C37H4gN4O4 +0.8H20 Calc: C, 70.85; H, 7.97; N, 8.93 Found: C, 71.12; H, 8.25; N, 8.45.

64) Compound 191 A nalysis: C 42 H 5l N 5 0 4 +.5CH2C12 C alc: C, 69.78; H, 7.16; N, 9.58 Found: C, 69.75; H, 7.31; N, 9.68.

65) Compound 203 A nalysis: C37H47N404 +l.lH20 C alc: C, 68.31; H, 7.62; N, 8.61 Found: C, 68.32; H, 7.57; N, 8.53.

66) Compound 204 A nalysis: C37H47ClN404 C alc: C, 68.66; H, 7.32; N, 8.66 Found: C, 68.32, H, 7.48; N, 8.42.

67) Compound 205 alysis: C 38 H 50 N 4 0 5 + 0.7 H 2 0 C alc: C, 69.63; H, 7.90; N, 8.54 Found: C, 69.72; N, 7.91; N, 8.54.

68) Compound 206 A nalysis: C 39 H 52 N 4 0 6 +0.8H20 C alc: C, 68.15; H, 7.86; N, 8.15 Found: C, 68.01; H, 8.02; N, 8.15.

W O 95/24207 PC~rrUS95102938 A. [S-(R*,S*)]3,3-Diethyl-2-[4-[[[2-(4-hydroxy-1-piperidinyl)ethyl]amino]carbonyl]phenoxy]-N-[ l -(4-methylphenyl)butyll -4-oxo- 1 -azetidinecarboxamide When 1-(2-aminoethyl)-4-benzyloxypiperidine is used in place of N,N,N-trimethylethylene ~ mine in the procedure of Example 1 lb there is obtained [S-(R*,S*)] 3,3-diethyl-2-[4-[[[2-(4-benzyloxy-1 -piperidinyl]ethyl]amino] carbonyl]phenoxy] -N- [1 -(4-methylphenyl)-butyl]-4-oxo- 1 -azetidinecarboxamide.

B. [S-(R+,S*)]-3,3-diethyl-2-[4-[[[2-(4-hydroxy-1-piperidinyl)ethyl]amino]carbonyl]phenoxy]-N-[ l -(4-methyl-phenyl)butyll-4-oxo- 1 -azetidine-carboxamide A solution of the amide from Step A above in 10 ml of glacial acetic acid cont~ining 22 mg of 10% Pd/C is hydrogenated under 42 lb hydrogen pressure. When TLC indicate completion of the reaction, the n~i~ is filtered and concentrated in vacuo after the addition of 50 ml toluene. The residue is dissolved in ethyl acetate, washed with saturated sodium bicarbonate solution. The organic layer is dried with sodium sulfate and concentrated in vacuo. The residue is chromatographed on 15 g silica gel using 5% methanol in methylene chloride and yields 96 mg of [S-(R+,S*)]-3,3-diethyl-2-[4-[[[2-(4-hydroxy- 1 -piperidinyl)ethyl]amino]carbonyl]phenoxy]-N-[ l -(4-methyl-phenyl)butyl]-4-oxo- 1 -azetidine-carboxamide.
Analysis: C33H46N4O5 +1.3H20 Calc: C,65.83;H,8.13;N,9.30 Found: C, 66.10; H, 8.06; N, 8.91.

When 1-(2-~minoethyl)-4-benzyloxypiperidine is replaced in the procedure of F.x~mple 12 by the appropriate amines, the following compounds 123, 124, 129, 131 and 138 are obtained, for example:

1) Compound 129 Analysis: C34H4gN4O5 Calc: C, 68.89; H, 8.16; N, 9.45 Found: C, 68.68; H, 8.18; N, 8.65.

2) Compound 131 Analysis: C32H44N405 +1H20 Calc: C, 65.92; H, 7.96; N, 9.61 Found: C, 66.07; H, 7.86; N, 9.45.
3) Compound 138 Analysis: C33H46N405 +0.5H20 Calc: C, 67.43; H, 8.06; N, 9.53 Found: C, 67.61; H, 8.06; N, 9.37.

Diamine Intermediates The ~ mines used to prepare the amino amides described herein were commercially available or prepared according to the following routes R, 1.) Cl-CH2CN R,~
R/ 2.) LAH ~N-C H2 C H2-N H2 A.N-cyanomethylhomopiperazine To a solution of 1.98 g homopiperazine in 50 ml acetone is added 4.25 g of powdered anhydrous sodium carbonate and 1.3 ml of 30 chloroacetonitrite. After 24 hrs the reaction mixture is filtered and the filter cake washed with 100 ml acetone. The combined filtrates are concentrated in vacuo and the residue chromatographed on silica gel using methylene chloride as the eluent. The yield of N-cyanomethyl homopiperazine is 2.69 g.

B.N-(2-aminoethyl)homopiperazine To a suspension of 1.02 g li~hiulll alllmimlm hydride in 50 ml of ether is slowly added a solution of 2.65 gm N-cyanomethyl homopiperazine in 25 ml ether. After the addition is complete the mixture is heated at reflux for 1 hour, then cooled to room temperature and quenched carefully with 1 ml water, 1 ml of 15% sodium hydroxide solution and 3 ml water. The mixture is filtered through sodium sulfate, the filter cake washed well with ether and the combined filtrates concentrated in vacuo to yield 2.60 g. N-(2-aminoethyl)homopiper-azine.
R~
/N-CH2CH2-NH2 1.) HCO2Et, NaOEt, R2 2.) BH3 THF
R1~
~N-CH2CH2-NH-CH3 N-(2-methylaminoethyl)homopiperazine A. N-(2-forrn~midoethyl)homopiperazine To a carefully prepared solution of 0.718 g of 60% sodium hydride dispension in 75 ml of absolute ethanol which has been cooled to 0C is added 7.3 ml of ethyl formate. After 5 minutes there is added a solution of 2.55 gm of N-(2-aminoethyl)homopiperazine in 25 ml - absolute ethanol. The mixture is stirred at room temperature overni~ht.
Saturated sodium bicarbonate solution (15 ml) is then added and the reaction mixture is stirred with 150 ml ethyl acetate, filtered through MgS04 and the filtrate concentrated in vacuo. Chromatography of the residue on 150 gm silica gel using an eluent of methylene chloride/methanol/conc. ammonium hydroxide (95/5/0.5) gives 2.78 g of N-(2-fonn~midoethyl)homopiperazine.

WO 95t24207 PCT/US95/02938 B. N-(2-methylaminoethyl)homopiperazine To a solution of 2.75 gm of N-(2-form~midoethyl)-homopiperazine in 20 ml of dry THF under N2 is added carefully 60 ml of borane THF solution. After the addition is complete the reaction mixture is heated to reflux for S hours then stirred at room temperature overnight. The reaction mixture is quenched by the careful addition of 20 ml of 6 N HCl followed by refluxing for 1 hour. The reaction mixture is cooled, 50 ml of water added and solid KOH added carefully to ~lk~line pH. Extraction with ether gives the desired product N-(2-methylamlno)homopiperazlne .

~CH2 CH3NH-CH2CH2-N\ 1.) R-X, Na2CO3 CH3 2.) H2lHoAcpd(oH)2c A. N-benzyl-N,N'-dimethyl-N'-(2-phenylethyl)ethylene-diamine A mixture of 0.900 gm N-benzyl-N,N'-dimethylethyl-ene(li~mine, 1.10 gm powdered sodium carbonate and 0.75 ml of 2-25 phenylethylbromide is refluxed for 5 hours. An additional 0.25 ml ofbromide is added during this time. The reaction mixture is then cooled and filtered. The filterate is concentrated in vacuo and the residue chromatographed on silica gel using an eluent of CH2C12/CH30H/-NH40H (97/3/0.3) to yield 0.875 gm of N-benzyl-N,N'-dimethyl-N'-(2-3 phenylethyl)ethylene~ mine.
.
B. N N'-dimethyl-N-(2-phenylethyl)ethylene~ mine To a solution 0.870 gm N-benzyl-N-N'-dimethyl-N'-(2-phenylethyl)ethylene~ mine in 10 ml ethanol and 5 ml acetic acid is added 0.18 gm Pd (OH)2/C. The mixtllre is hydrogenated at 40 psi for WO 95/24207 PCT/US95~'~,Z538 3.5 hours, then filtered and concentrated in vacuo. The residue is made akaline with lN NaOH and extracted well with ethyl acetate (5 X 25 ml). The combined extracts are filtered through sodium sulfate and 5 concentratedto yield N,N'-dimethyl-N-(2-phenylethyl)ethylene~ mine.

1.) Ar-CHO

o R ~ 2.) LAH

Rl~
N-CH2CH2-NH-CH2-Ar R2~

A solution of 1.28 gm 1-(2-amino-ethyl)piperdine and 1.07 gm pyridine-3-carboxaldehyde in 40 ml of toluene is heated to reflux under a Dean Stark trap. After 10 ml toluene distilled over the NMR of an aliquot indicated no aldehyde left. The reaction mixture was concentrated and the imine used directly in the next step.

B.
To a suspension of 0.380 gm of lithium ~ minum hydride in 30 ml of dry THF which has been cooled to -10C is added dropwise a solution of the above imine in 20 ml of dry THF. After about 1 hour the cold reaction mixture is quenched by the addition of S ml of 5 N
NaOH, then diluted with 100 ml ether and 20 ml of water. The organic layer is separated, washed with brine, filtered thru sodium sulfate and concentrated to give 2.17 gm of 1-[2-(3-pyridylmethylamino)ethyl]-piperidine suitable for use in subsequent reactions.

A,CH2CI

f H2 Ar To 7.50 gm of N,N'-dimethylethylene~ mine which has been cooled in an ice-ethanol bath is added portionwise over a 30 minute period 1.40 gm of 3-picolyl chloride. After stirring cold for 1 hour after the addition is completed, the reaction mixture is concentrated in vacuo and the residue partitioned between 50 ml of 5 ether and 10 ml of 5 N NaOH solution. The organic layer is separated and the aqueous layer extracted 2 times with 50 ml of ether. The combined organic extracts are dried through sodium sulfate and concentrated in vacuo. Chromatography on 150 gm silica gel using CH2C12/CH30H/NH40H (90/10/1) as eluent gives 0.930 g of N,N'-2 dimethyl-N-(3-pyridylmethyl)ethylene~ mine.

Amino Acid ~ diamine A. To an ice cooled solution of 2.29 N-CBZ-D-Proline in 50 ml of CHCl2 is added 1.35 gm 1-hydroxybenzotriazole hydrate followed by 2.06 gm of dicyclohexylcarbodiimide. After 20 lllillU~t~S, 0.85 ml of pyrrolidine is added and the reaction mixture stirred 3 overnight after which time it is filtered and the filtrate concentrated in vacuo. The residue is partitioned between 100 ml ethyl acetate and 50 ml of 2 N hydrochloric acid. The organic layer is separated, washed with 50 ml of 1.0 N sodium hydroxide solution, dried through sodium sulfate and concentrated in vacuo. Chromatography on 150 gm of silica WO 95/24207 PCT/USÇ5/'~93~
-- 2 1 843&5 gel using ethylacetate in hexanes (30-100%) as eluent gives 2.04 gm of the desired pyrrolidine amide 5 B- To a solution of 1.519 gm of the amide (prepared in A) in 20 ml absolute ethanol is added 75 mg of 10% Pd on carbon catalyst.
The mixture is hydrogenated at 40 psi for about an hour then filtrate and the filtrate concentrated to yield D-proline pyrrolidine amide.

o C To a suspension of 0.380 gm of lithium ~hlminum hydride in 15 ml of dry tetrahydrofuran is carefully added a solution of the D-proline amide (prepared in B above) in 10 ml tetrahydrofuran. The mixture is refluxed for 2 hrs then cooled and quenched with 2 ml of 2.5 N sodium hydroxide. The mi~ul~; is filtered through a pad of sodium sulfate and the filter cake washed with 2 x 50 ml of ether. The combined filtrates are concentrated in vacuo to yield 0.80 gm of desired 2-( 1 -pyrrolidinylmethyl)pyrrolidine.

[S-(R*,S*)]-2-[4-[[(4-Methyl)piperazin-1-yl]carbonyl] phenoxy]-((3 ,3-diethyl-N-[ 1 -(methylphenyl)butyl] -4-oxo- 1 -azetidinecarbox-amide A solution of S-(R*,S*)]-4-(((3,3-diethyl-1- ((4-methyl-2 5 phenyl)butylamino)carbonyl)-4-oxo-2-azetidinyl)oxy)benzoyl chloride (3.8 mmol), prepared as in Example 1 lA, in 50 ml of methylene chloride was cooled in an ice bath and a solution of 0.70 gm of N-methylpiperazine in 10 ml of methylene chloride was added over 5 min.
The reaction was stirred for 1 hr and was then poured into a mixture of 30 ice water and 10% potassium carbonate. The product was extracted with two portions of methylene chloride and each methylene chloride layer was washed with a portion of brine. The methylene chloride layers were combined, dried over sodium sulfate and evaporated. The residue was purified with flash chromatography using ethyl acetate, then W O 95/24207 PCTrUS95/02938 21 84~85 2% triethyl~mine/10% methanoV88% ethyl acetate to afford 2.1 gm of the title compound as a white solid.
Analysis: C30H42N4O4 Calc: C, 69.64; H, 7.92; N, 10.48 Found: C, 69.62; H, 8.23; N, 10.46.

[S-(R*,S*)]-2-[4-[[(4-Methyl)piperazin-1-yl]carbonyl] phenoxy]-((3,3-diethyl-N- [1 -(3,4-methylenedioxyphenyl)butyl] -4-oxo- 1 -azetidine-carboxamide When [S-(R*,S*)]-4-(((3,3-diethyl-1-((3,4-methylene-dioxyphenyl)butylamino)carbonyl)-4-oxo-2-azetidinyl)oxy)benzoyl chloride (3.1 mmol), prepared as in Example 1 lA, was reacted with N-methylpiperazine as in Example 19, there was obtained 1.75 gm of the title compound.
Analysis: C31H40N406 Calc: C, 65.94; H, 7.14; N, 9.92 Found: C, 65.80; H, 7.31; N, 10.05.

[S-(R*,S*)]-2-[4-[[(4-Hydroxyethyl)piperazin-1-yl] carbonyl]phenoxy]-((3 3 -diethyl-N- r 1 -(methylphenyl) butyll -4-oxo- 1 -azetidinecarboxamide When [S -(R*,S *)] -4-(((3,3 -diethyl- 1 -((4-methylphenyl)-butylamino)carbonyl)-4-oxo-2-azetidinyl)oxy)benzoyl chloride (3.8 mmol), ~r~lJaled as in F.x~mple 11A, was reacted with N-(2-hydroxy-ethyl)piperazine (7.6 mmol) and diisopropylethyl~mine (3.8 mmol) as in Fx~mrle 19, there was obtained 2.1 gm of the title compound.
Analysis: C32H44N4O5 Calc: C, 68.06; H, 7.85; N, 9.92 Found: C, 67.88; H, 7.87; N, 10.17.

21 843~5 [S-(R*,S*)]-2-[4-[[(4-Hydroxyethyl)pil~efazin-l-yl] carbonyl]phenoxy]-((3 ,3 -diethyl-N- [ 1-(3 ,4-methylenedioxyphenyl)butyl] -4-oxo- 1-azetidinecarboxamide When [S-(R*,S*)]-4-(((3,3-diethyl-1-((3,4-methylenedioxy-phenyl)butylamino)carbonyl)-4-oxo-2-azetidinyl)oxy)benzoyl chloride (3.1 mmol), prepared as in Fx~mple llA, was reacted with N-(2-hydroxyethyl)piperazine (6.2 mmol) and diisopropylethyl~mine (3.1 mmol) as in Example 19, there was obtained 1.50 gm of the title compound.
Analysis: C32H42N407-1.5H20 Calc: C, 61.94; H, 6.89; N, 9.06 Found: C, 61.95; H, 6.92; N, 8.96.

[S-(R*,S*)]-2-[4-[[(4-Cyclopropyl)~i~e~dzin-l-yl] carbonyl]phenoxy]-((3 ,3-diethyl-N-[ 1 -(4-methylphenyl)butyl]-4-oxo- 1 -azetidine-carboxamlde To a solution of [S-(R*,S*)]-4-(((3,3-diethyl-1-((4-methyl-phenyl)butylamino)carbonyl)-4-oxo-2-azetidinyl)oxy)benzoyl chloride (3.8 mmol), prepared as in Fx~mple 1 lA, in 50 ml of methylene chlorine was added N-(cyclopropyl)piperazine dihydrochloride (5.7 mmol) and then a solution of diisopropylethyl~min~ (15.8 mmol) in 10 ml of methylene chloride was added over 5 min with ice-bath cooling.
The reaction was stirred for 1 hr at 0C and then poured into ice water.
The product was extracted with two portions of methylene chloride and each methylene chloride layer was washed with a portion of brine. The methylene chloride layers were combined, dried over sodium sulfate and ethyl acetate/50% hexanes, then 70% ethyl acetate/30% hexanes to afford 2.1 gm of the title compound as a white solid.

Analysis: C32H42N404 Calc: C, 70.69; H, 7.91; N, 9.99 Found: C, 70.62; H, 8.04; N, 9.95.

[S-(R*,S*)]-2-[4-[[(4-Cyclopropyl)piperazin-1-yl] carbonyl]phenoxy]-((3 ,3-diethyl-N- [ 1 -(3 ,4-methylenedioxyphenyl)butyl] -4-oxo- 1-azetidinecarboxamide o When [S-(R*,S*)]-4-(((3,3-diethyl-1 -((3,4-methylene-dioxyphenyl)butylamino)carbonyl)-4-oxo-2-azetidinyl)oxy)benzoyl chloride (3.1 mmol), ~r~ared as in Example 1 lA, was reacted with N-(cyclopropyl)piperazine (4.6 mmol) and diisopropylethylamine (9.3 mmol) as in Example 23, there was obtained 1.80 gm of the title compound.
Analysis: C32H42N407 Calc: C, 67.10; H, 7.17; N, 9.49 Found: C, 67.03; H, 7.31; N, 9.47.

[S-(R*,S*)]-2-[4-[[(4-Piperazin-1 -yl)carbonyl]phenoxy]-((3,3-diethyl-N-r 1 -(4-methylphenyl)butyll -4-oxo- 1 -azetidinecarboxamide Step A: [S-R*,S*)]-2-[4-[[(4-(t-Butoxycarbonyl))piperazin-1-yl]carbonyl]phenoxy] -((3 ,3-diethyl-N-[ 1 -(4-methyl-phenyl)butyll -4-oxo- 1 -azetidinecarboxamide [S-(R*,S *)] -4-(((3 ,3 -Diethyl- 1 -((4-methylphenyl)-30 butylamino)carbonyl)-4-oxo-2-azetidinyl)oxy)benzoyl chloride (0.4 mmol), prepared as in Fx~mple 1 lA, was reacted with N-(t-butoxy-carbonyl)piperazine (0.6 mmol) and triethyl~mine (1.2 mmol) as in Fx~mple 23. The crude, title product so obtained was used directly in the following Step B.

WO 9~/24207 PCT/US95/02938 Step B: [S-R*,S*)]-2-[4-[(Piperazin-1-yl)carbonyl]phenoxy]-((3,3-diethyl-N-[1 -(4-methylphenyl)butyl-4-oxo- 1 -azetidine-carboxamide The product from Step A was dissolved in 0.5 ml of anisole and 2 ml of cold TFA was added. The reaction was stirred at 0C for 1 hr and was then diluted with methylene chloride and evaporated. The residue was taken up in methylene chloride, washed with 10% sodium carbonate and brine, dried over sodium sulfate and concentrated. The o residue was purified by flash chromatography using 5, then 10%
methanol/methylene chloride to afford 0.212 gm of title product.
Analysis: C30H40N4os-lH2o Calc: C, 66.89; H, 7.85; N, 10.40 Found: C, 67.06; H, 7.55; N, 10.30.

[S-(R*,S *)]-2-[4-[ [(4-Piperazin- 1 -yl)carbonyl]phenoxy] -((3,3-diethyl-N-~ 1 -(3.4-methylenedioxyphenyl)butyll-4-oxo-1 -azetidinecarboxamide Step A: [S-R*,S*)]-2-[4-[[(4-Benzyloxycarbonyl) Piperazin-1-yl]carbonyl]phenoxy]-((3,3-diethyl-N-[1 -3,4-methylene-dioxyphenyl)butyll-4-oxo- 1 -azetidinecarboxamide When [S-(R*,S*)]-4-(((3,3-diethyl-1-((3,4-methylene-2 5 dioxyphenyl)butylamino)carbonyl)-4-oxo-2-azetidinyl)oxy)benzoyl chloride (0.41 mmol), prepared as in Example llA, was reacted with N-(benzyloxycarbonyl)piperazine (0.77 mmol) and diisopropyl-ethyl~mine (1.6 mmol) as in Fx~mple 23, there was obtained 290 mg of the title compound.

Step B: [S-R*,S*)]-2-[4-[(Piperazin-1-yl)carbonyl]phenoxy]-((3,3-diethyl-N-[1 -(3,4-methylenedioxyphenyl)butyl-4-oxo- 1 -azetidinecarboxamide A solution of 250 mg of material from Example 26, Step A
in 10 ml of ethanol was hydrogenated at 40 p.s.i. over 50 mg of 10%

Pd/C for 16 hrs. The reaction was filtered and evaporated. The residue was purified by preparative TLC eluting with 2% TEA/10% methanol/-88% ethyl acetate to afford 150 mg of title product.
Analysis: C30H38N4o6-3H2o Calc: C, 59.59; H, 7.33; N, 9.29 Found: C, 59.66; H, 7.65; N, 9.61.

Claims (11)

WHAT IS CLAIMED IS:

1. A pharmaceutical composition comprising a therapeutically effective, non-toxic amount of the (F)-actin shortening protein gelsolin and a therapeutically effective amount of a compound of Formula (I):

or a pharmaceutically acceptable salt thereof wherein:
R is C1-6alkyl;
R1 is C1-6alkyl or C1-6alkoxy-C1-6alkyl;
M is (1) hydrogen, (2) C1-6alkyl, (3) hydroxy C1-6alkyl, (4) halo C1-6alkyl, (5) C2-6alkenyl, or (6) C1-6alkoxy-C1-6alkyl;
Ra and Rb are each individually hydrogen;
R2 and R3 are each independently (1) hydrogen, (2) C1-6alkyl, (3) halo, or (4) C1-6alkoxy.
or R and R3 are joined together to form a methylenedioxy group or a furan ring, R4 is wherein Q is a covalent bond Y is R12 R10 -N- C n-C-R9, R10, R11 and R12 are each individually hydrogen or C1-3alkyl;
R7 and R8 are each individually (a) hydrogen, (b) C1-6alkyl, (c) C1-6alkyloxy C2-3alkyl, (d) hydroxy C2-6alkyl, or n is 1,2,3,4 or 5;

R8 and R9 are joined together to form a mono or di substituted saturated monocyclic ring of 6 to 7 atoms and having two hetero atoms which are the nitrogens to which R8 and R9 are attached; and the substituents are independently selected from hydrogen and C1-3alkyl.

2. A composition according to Claim 1 wherein R is C1-3alkyl;
R1 is C1-3alkyl or C1-3alkoxy-C1-3alkyl;
M is (1) hydrogen, (2) C1-3alkyl, (3) C2-3alkenyl, or Ra and Rb are each hydrogen;
R and R3 are each independently (1) hydrogen, (2) C1-3alkyl, (3) C1-3alkoxy, R and R3 are joined together to form a methylenedioxy group or a furan ring;

R4 is , wherein Q is a covalent bond;

Y is R9, R10, R11 and R12 are each individually hydrogen or C1-2alkyl;
R7 and R8 are each individually (a) hydrogen, (b) C1-3alkyl, (c) C1-3alkyloxy C2-3alkyl, n is 1,2,3 or 4;
R8 and R9 are joined together to form a mono or di substituted saturated monocyclic ring of 6 to 7 atoms and having two hetero atoms which are the nitrogens to which R8 and R9 are attached; said rings selected from piperazinyl and homopiperazinyl; and the substituents are independently selected from hydrogen and C1-3alkyl.

3. A composition according to Claim 2 wherein R is C1-3 alkyl;
R1 is C1-3 alkyl;
M is (a) C1-3 alkyl, or (b) C2-3 alkenyl;
R is (a) hydrogen (b) C1-3 alkyl and R3 is hydrogen, or R and R3 are joined together to form a methylenedioxy group or a furan ring;
R7 and R8 are each independently selected from (a) hydrogen.
(b) C1-2alkyl, (c) C1-2 alkoxy C2-3 alkyl, R8 and R9 are joined together to form a mono or di substituted saturated monocyclic ring of 6 to 7 atoms and having two hetero atoms which are the nitrogens to which R8 ant R9 are attached; said rings selected from piperazinyl and homopiperazinyl; and the substituents are independently selected from hydrogen and C1-3alkyl.

4. A composition according to Claim 3 wherein R is methyl or ethyl;
R1 is methyl or ethyl;
M is (a) methyl a ethyl, or (b) allyl;
R and R3 are each hydrogen, or R ant R3 are joined together to form a methylenedioxy group or a furan ring;
n is 1 or 2;
R8 and R9 are joined together to form a mono or di substituted saturated monocyclic ring of 6 to 7 atoms and having two hetero atoms which are the nitrogens to which R8 and R9 are attached; said rings selected from piperazinyl and homopiperazinyl; and the substituents are independently selected from hydrogen and C1-3alkyl.

5. A pharmaceutical composition comprising a therapeutically effective, non-toxic amount of the (F)-actin shortening protein gelsolin, a pharmaceutically acceptable carrier and a therapeutically effective amount of a compound of Formula (II):

II

wherein R is:

(1) -CH3, (2) 4-fluorophenyl, (3) 3-chlorophenyl, (4) phenyl, (5) benzyl, (6) H, (7) i-Pr, (8) i-Bu, (9) -CH2CO2Et, (10) -CH2CO2H, (11) Et, (12) Pr, (13) 2-pyrimidinyl (14) -CH2CH2OC(O)NHCH3, (15) cyclopropyl, or (16) -CH2CH2OH.

6. A composition according to Claim 5 wherein the compound of Formula I is [S-(R*,S*)]-2-[4[[(4-methyl)piperazin -1-yl]carbonyl]phenoxy]-3,3-diethyl-N-[1-(3,4-methylenedioxyphenyl)butyl]-4-oxo-1-azetidinecarboxamide.

7. A method of treating lung disease in a patient in need of such treatment comprising:
administration to a patient in need of such treatment a composition according to Claim 2.

8. A method of treating lung disease in a patient in need of such treatment comprising:
administration to a patient in need of such treatment a composition according to Claim 3.

9. A method of treating lung disease in a patient in need of such treatment comprising:
administration to a patient in need of such treatment a composition according to Claim 6.

10. A method of treating a patient with a lung disease, comprising;
administration to a patient in need of sputum viscosity reduction, a composition according to Claim 6, wherein said amounts are effective to return the lung function of said patients to at least 60-75% of normal as measured by FEV1.

11. A method of treating a patient with a lung disease, comprising;
administration to a patient in need of sputum viscosity reduction, a composition according to Claim 6, wherein said amounts are effective to return the lung function of said patients to at least 75-90% of normal as measured by FEV1.