Sep 10, 1997 - ethanesulfonate, fumarate, glucoheptanoate, glycerophosphate ... salts, alkali metal salts such as sodium and potassium salts, alkaline earth metal salts such ... chain halides such as decyl, lauryl, myristyl and stearyl chlorides ...
US005872138A
United States Patent [19]
[11]
Patent Number:
Naylor-Olsen et al.
[45]
Date of Patent:
[54]
THROMBIN INHIBITORS
[75]
Inventors: Adel M. Naylor-Olsen; Gerald S. P0nticell0, both of Lansdale; Joseph P.
1990.
Archibald J1 and Benke GA. J Med. Chem. 17 (7), pp.
Lansdale; Craig Coburn, Skippack;
736—739, 1974.
Brian T. Phillips, Telford; S. D. Lewis, Lansdale; Mark E. Fraley, North
Mack et al., J. Enzyme Inhibition, “Design, Synthesis and
Wales, all of Pa.
Biological Activity of Novel Rigid Amidino—Phenylala nine
[73] Assignee: Merck & Co., Inc., Rahway, NJ.
[51] [52]
Int. Cl.6 ....................... .. A61K 31/44; C07D 213/72 US. Cl. ........................ .. 514/352; 514/255; 514/637;
[58]
Field Of Search .......................... .. 546/312; 544/383;
544/383; 546/312; 564/245 564/245; 514/352, 255, 637
.
. ”, vol. 9, pp. 73—86 (1965).
Bernstein, et al., J. Med. Chem., 37, 3313—3326 “Nonpep
Primary Examiner—John Kight Assistant Examiner—Evelyn Huang Attorney, Agent, or Firm—Richard S. Parr; Melvin Winokur
[57]
ABSTRACT
A compound Which inhibits human thrombin and Which has the general structure
References Cited US. PATENT DOCUMENTS
J —A
1/1992 Alig ...................................... .. 514/353
5,260,307 11/1993 Ackermann et al. 5,405,854 4/1995 Ackermann et al.
514/323 514/315
5,459,142 5,510,369 5,518,735 5,612,363
514/252 514/422 424/449 514/392
10/1995 4/1996 5/1996 3/1997
Tonee et a1. ....... .. Lumma et a1. ........ .. SturZebecher et al. ............... .. Mohan et al. ........................ ..
FOREIGN PATENT DOCUMENTS 0 262 096
.
tidic Inhibitors of Human Leukocyte Elastase . . . ”, 1994.
[21] Appl. No.: 926,606 [22] Filed: Sep. 10, 1997
5,084,466
Feb. 16, 1999
Efange MN et al. J. Med. Chem. 33 (12), pp. 3133—3138,
Vacca, Telford; Randall W. Hungate,
[56]
5,872,138
3/1988
European Pat. Off. .
0 509 769 A2 10/1992
European Pat. Off. .
94/25051 11/1994 96/11697 4/1996 96/31504 10/1996 96/32110 10/1996 97/01338 1/1997
WIPO . WIPO . WIPO . WIPO . WIPO .
X
R1\(
R2
R3 I
such as
:
CH3
O@/
NH
N
OTHER PUBLICATIONS
/
Johnson RA et al. J. Med Chem. 29(8), pp. 1461—1468, 1986. Cohen T and Deets GL. J. Am. Chem. Soc. 94 (3), pp. 932—937, Feb. 1972.
| \
13 Claims, N0 Drawings
5,872,138 1
2
THROMBIN INHIBITORS
blood, blood products, or mammalian organs in order to effect the desired inhibitions.
This application claims bene?t of the provisional appli cation 60/026,033, ?led on Sep. 13, 1996. BACKGROUND OF THE INVENTION
Thrombin is a serine protease present in blood plasma in the form of a precursor, prothrombin. Thrombin plays a
The invention also includes a composition for preventing
5 or treating unstable angina, refractory angina, myocardial infarction, transient ischemic attacks, atrial ?brillation, thrombotic stroke, embolic stroke, deep vein thrombosis, disseminated intravascular coagulation, ocular build up of ?brin, and reocclusion or restenosis of recanaliZed vessels, in a mammal, comprising a compound of the invention in a
central role in the mechanism of blood coagulation by
converting the solution plasma protein, ?brinogen, into
pharmaceutically acceptable carrier. These compositions may optionally include anticoagulants, antiplatelet agents, and thrombolytic agents.
insoluble ?brin.
EdWards et al., J. Amer. Chem. Soc., (1992) vol. 114, pp. 1854—63, describes peptidyl a-ketobenZoXaZoles Which are reversible inhibitors of the serine proteases human leukocyte
The invention also includes the use of a compound of the invention in the manufacture of a medicament for preventing
elastase and porcine pancreatic elastase.
or treating unstable angina, refractory angina, myocardial infarction, transient ischemic attacks, atrial ?brillation, thrombotic stroke, embolic stroke, deep vein thrombosis,
European Publication 363 284 describes analogs of pep tidase substrates in Which the nitrogen atom of the scissile
amide group of the substrate peptide has been replaced by hydrogen or a substituted carbonyl moiety. Australian Publication 86245677 also describes peptidase inhibitors having an activated electrophilic ketone moiety such as ?uoromethylene ketone or ot-keto carboXyl deriva tives. Thrombin inhibitors described in prior publications con tain sidechains of arginine and lysine. These structures shoW loW selectivity for thrombin over other trypsin-like enZymes. Some of them shoW toXicity of hypotension and
disseminated intravascular coagulation, ocular build up of ?brin, and reocclusion or restenosis of recanaliZed vessels, in a mammal.
The invention also includes a method for reducing the thrombogenicity of a surface in a mammal by attaching to the surface, either covalently or noncovalently, a compound of the invention. DETAILED DESCRIPTION OF THE
liver toXicity. European Publication 601 459 describes sulfonamido 30 heterocyclic thrombin inhibitors, such as N-[4
[(aminoiminomethyl)amino]butyl]-1 —[N-(2 naphthalenylsulfonyl)-L-phenylalanyl]-L-prolinamide.
INVENTION
Compounds of the invention have the folloWing structure: J —A
WO 94/29336 describes compounds Which are useful as thrombin inhibitors.
R1\(
SUMMARY OF THE INVENTION
R2
A compound Which inhibits human thrombin and Which has the general structure
40
I
R3
and pharmaceutically acceptable salts thereof Wherein X is
I —A
—N(R4 —,
X
R1\(
R2
45
R3 I
such as
:
CH3
O@/
NH
| \ N /
The invention includes a composition for inhibiting loss 60
of blood platelets, inhibiting formation of blood platelet aggregates, inhibiting formation of ?brin, inhibiting throm bus formation, and inhibiting embolus formation in a mammal, comprising a compound of the invention in a
pharmaceutically acceptable carrier. These compositions may optionally include anticoagulants, antiplatelet agents, and thrombolytic agents. The compositions can be added to
R1, R2, and R4 are independently selected from
hydrogen, aryl C1_4 alkyl, diaryl C1_4 alkyl, dicyclo C3_8 alkyl C1_4 alkyl, cyclo C3_8 alkyl C1_4 alkyl,
5,872,138 4 R1 and R2 are independently selected from R5 —CON
hydrogen, aryl, or cyclo C3_7 alkyl unsubstituted or substituted With one
,
or more of C1_4 alkyl, C1_4 alkoXy, benZyl, cyclo heXylrnethyl or aryl;
wherein R5 and R6 are independently hydrogen or
CL4 alkyl,
R3 is
substituted aryl With one or tWo substituents selected
from
1O
C1-4 alkyl, C1_4 alkoXy,
C1_4 alkyl, or
C1_4 alkenyl;
R5 —CON
hydrogen,
A is selected from one of the folloWing fragments ,
15
Wherein R5 and R6 are independently hydrogen or C1_ 4 alkyl,
aryloXy, cyclo C3_8 alkoXy, rnethylenedioXy,
HZN
halogen, or
hydroXy,
6 \ ,
N— ,
or
EN
—
5
25
heteroaryl With one or tWo heteroatorns selected from
N, O, and S, cyclo C3_7 alkyl unsubstituted or
One class of this group of compounds has the folloWing
substituted With one or more of CL4 alkyl, CL4
structure:
alkoXy, benZyl, cycloheXylrnethyl or aryl, a C4_1O carbocyclic or bicyclic ring, or R1 and R2 along With the carbon to Which they attach form
J-A
a cyclo C3_7 alkyl ring; R3 is
hydrogen, C1-4 alkyl,
O
RLY 35
C1_ 4 alkenyl,
C1_4alkoXy, —NHR7 Wherein R7 is hydrogen or CL4 alkyl, or
and pharrnaceutically acceptable salts thereof Wherein
—NHSO2CH2aryl; A is selected from one of the folloWing fragments
J is CH2, CHZNH, or S02; R1 and R2 are independently selected from 45
hydrogen, aryl, or
cycloheXyl; R3 is
hydrogen, —CH=CH2or —CH2CH3; and A is selected from one of the folloWing fragrnents 55
One group of compounds of the invention have the
folloWing structure:
HN
HZN
and pharrnaceutically acceptable salts thereof Wherein J is (CH2)m, (CH2)mNH, or S02, Where In is 1 or 2;
Speci?c eXernpli?cations of this class are
5,872,138 6 -continued
Designation D—3,3—dicha
D-3,3-Dicyclohexylalanine
Pro
Proline
Arg Gly D—3,3,—diphe
Arginine Glycine D—3,3—Diphenylalanine
The compounds of the present invention may have chiral centers and occur as racemates, racemic mixtures and as
individual diastereomers, or enantiomers With all isomeric
forms being included in the present invention. 15
When any variable occurs more than one time in any constituent or in formula I, its de?nition on each occurrence
is independent of its de?nition at every other occurrence. Also, combinations of substituents and/or variables are permissible only if such combinations result in stable com
pounds. As used herein, except Where noted, “alkyl” is intended to include both branched- and straight-chain saturated aliphatic hydrocarbon groups having 1—8 carbon atoms (Me is methyl, Et is ethyl, Pr is propyl, Bu is butyl). “Alkenyl” is 25
intended to include both branched- and straight-chain unsat
urated aliphatic hydrocarbon groups having 1—8 carbon atoms, e.g. ethenyl, propenyl, etc. “Cycloalkyl” includes cyclic saturated aliphatic hydrocarbon groups having 3—8 carbon atoms. “Aryl” means a 6-membered organic radical derived from an aromatic hydrocarbon by removal of one hydrogen atom. “Heteroaryl” means a 5- or 6-membered
organic radical having 1 or 2 heteroatoms selected from N,
O, and S. “Alkoxy” represents an alkyl group having 1—8 35
carbon atoms attached through an oxygen bridge. “Halo”, as used herein, means ?uoro, chloro, bromo and iodo. “Coun
terion” is used to represent a small, single negatively
charged species, such as chloride, bromide, hydroxide,
acetate, tri?uroacetate, perchlorate, nitrate, benZoate, maleate, tartrate, hemitartrate, benZene sulfonate, and the like. Under standard nonmenclature used throughout this dis
45
closure unless speci?ed otherWise, the terminal portion of the designated side chain is described ?rst folloWed by the adjacent functionality toWard the point of attachment. For example, an ethyl substituent substituted With “methylcar
bonylamino” is equivalent to
and pharmaceutically acceptable salts thereof. ABBREVIATIONS
Designation BOC (Boc) HBT(HOBT or HOBt) BBC reagent
t-butyloxycarbonyl 1-hydroxybenzotriazole hydrate benzotriazolyloxy-bis(pyrrolidino)— carbonium hexa?uorophosphate
PyCIU
1,1,3,3—bis(tetramethylene)—
EDC
chlorouronium hexa?uorophosphate 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride
The term heterocycle or heterocyclic, as used herein except Where noted, represents a stable 5- to 7-membered mono- or bicyclic or stable 7- to 10-membered bicyclic 55
heterocyclic ring system any ring of Which may be saturated or unsaturated, and Which consists of carbon atoms and from one to three heteroatoms selected from the group consisting
(BOC)2O
di-t-butyl dicarbonate
DMF
dimethylformamide
Et3N or TEA EtOAc
triethylamine ethyl acetate
of N, O and S, and Wherein the nitrogen and sulfur heteroa toms may optionally be oxidized, and the nitrogen heteroa tom may optionally be quatemiZed, and including any bicy clic group in Which any of the above-de?ned heterocyclic rings is fused to a benZene ring. The heterocyclic ring may
TFA
tri?uoroacetic acid
DMAP DME
dimethylaminopyridine dimethoxyethane
be attached at any heteroatom or carbon atom Which results
BH3-THF
Borane-tetrahydrofuran complex
D—Phe(3,4—Cl2)
D-3,4-Dichlorophenylalanine
in the creation of a stable structure. Examples of such 65
heterocyclic elements include piperidinyl, piperaZinyl,
2-oxopiperaZinyl, 2-oxopiperidinyl, 2-oxopyrrolodinyl, 2-oxoaZepinyl, aZepinyl, pyrrolyl, 4-piperidonyl,
5,872,138 7 pyrrolidinyl, pyraZolyl, pyraZolidinyl, imidaZolyl, imidaZolinyl, imidaZolidinyl, pyridyl, pyraZinyl, pyrimidinyl, pyridaZinyl, oxaZolyl, oxaZolidinyl, isoxaZolyl, isoxaZolidinyl, morpholinyl, thiaZolyl, thiaZolidinyl, isothiaZolyl, quinuclidinyl, isothiaZolidinyl, indolyl, quinolinyl, isoquinolinyl, benZimidaZolyl, thiadiaZoyl, benZopyranyl, benZothiaZolyl, benZoxaZolyl, furyl, tetrahydrofuryl, tetrahydropyranyl, thienyl, benZothienyl, thiamorpholinyl, thiamorpholinyl sulfoxide, thiamorpholi nyl sulfone, and oxadiaZolyl. Morpholino is the same as
8 SCHEME 1 OH
Cs2CO3/DMF CH Br
CO2CH2CH3 10
R1
M
1-2
morpholinyl. The pharmaceutically-acceptable salts of the compounds of Formula I (in the form of Water- or oil-soluble or
dispersible products) include the conventional non-toxic
15
salts or the quaternary ammonium salts Which are formed,
R1 0
e.g., from inorganic or organic acids or bases. Examples of
such acid addition salts include acetate, adipate, alginate,
LAH
THF>
aspartate, benZoate, benZenesulfonate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate,
cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptanoate, glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, oxalate, pamoate, pectinate, persulfate, 3-phenylpropionate, picrate, pivalate, propionate, succinate, tartrate, thiocyanate,
CO2CH2CH3 1-3
25
R1
tosylate, and undecanoate. Base salts include ammonium salts, alkali metal salts such as sodium and potassium salts, alkaline earth metal salts such as calcium and magnesium salts, salts With organic bases such as dicyclohexylamine
0
THF/CBr4
(C5H5)3P E
salts, N-methyl-D-glucamine, and salts With amino acids such as arginine, lysine, and so forth. Also, the basic
nitrogen-containing groups may be quaterniZed With such agents as loWer alkyl halides, such as methyl, ethyl, propyl,
35
CHZOH
and butyl chloride, bromides and iodides; dialkyl sulfates
1-4
like dimethyl, diethyl, dibutyl and diamyl sulfates, long chain halides such as decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides, aralkyl halides like benZyl
2 R1
and phenethyl bromides and others. Amide couplings used to form the compounds of this invention are typically performed by the carbodiimide method With reagents such as dicyclohexylcarbodiimide, or
45
O
4-aminopyridine 5
1-ethyl-3-(3-dimethylaminopropyl) carbodiimide. Other methods of forming the amide or peptide bond include, but are not limited to the synthetic routes via an acid chloride,
aZide, mixed anhydride or activated ester. Typically, solution
phase amide coupling are performed, but solid-phase syn thesis by classical Merri?eld techniques may be employed Br
instead. The addition and removal of one or more protecting I 5
groups is also typical practice. Compounds of the invention can be prepared according to the general procedures outlined in Schemes I—VI.
55
Compounds found in table 1 Were synthesiZed by the general method outlined in Scheme I and are exempli?ed by, but not limited to the examples 1—9. A phenolic ester such as ethyl 4-hydroxybenZoic acid (1) is alkylated With an alkyl
R1 0
bromide such as benZyl bromide by using a base such as cesium carbonate to afford the product 1-3. The ester group is then reduced With a hydride reducing agent such as
lithium aluminum hydride to afford the benZylic alcohol 1-4. The benZylic alcohol is converted into a leaving group (such
I-6
65
A general method for synthesiZing compounds found in
as a bromide or tri?ate) and the benZylic leaving group is
tables 2 and 3 is outlined Scheme II. AbenZoic acid such as
reacted With 4-aminopyridine to afford the ?nal product.
II-1 (obtained in standard fashion) is coupled to 4-amino
5,872,138 9
10
pyridine under standard amide bond coupling conditions. The resultant arnide 11-2 is then reduced With lithium alu
minum anhydride to afford the desired product II-3. SCHEME II
< > \
O
EDC/HOBT
COZH
EtgN/DMF 4-AP
11-1
1) (COCl)2—CH2Cl2 Et3N—DMF 2) 4-AP
O O NH II-2
/ I \
N
LiAlH4/1‘HF 60° H
_
N
N
o
\
/
11-3
-
-
-
-
-
-
-
-
A general method for synthesizing cornpounds found in table 2 is outlined Schernes II and III. AbenZoate ester such as III-4 (obtained in standard fashion) is reacted With an
With an aqueous base and is coupled to 4-arn1nopyr1d1ne under Standard amide bond Coupling conditions_ The result ant amide 111-7 is then reduced With lithium aluminum
alkylating reagent (such as brornornethyl cycloheXane) to afford O-alkylated product III-5. The ester is hydrolyzed
hydride to afford the desired product IH_8_
SCHEME III
HO
/\/ o
/\/4 Br 3 Cs2CO3/DMF
/
230° c
COOCH3
>
COOCH3
III-1
III-2
Br HO
HO
H2/Pd(C) EtOAc /
Cs2CO3/DMF
COOCH3
COOCH3
III-3
III-4
o
W
NaOH/MeOH ; COOCH3 III-5
>
5,872,138 11
12 -continued SCHEME III 0
1. (C1CO)Z/DCE 2.
_
>
COOH
HZN
III-6
N
\
/
WO 0
%
III-7
III-8
SCHEME IV Br HO
0
BrZ/Fe/HCOOH
Cs2CO3/DMF
>
100° c.
coocH3
>
coocH3
IV-1
W2 0
o
W
Pd(PPh3)4/dioXane > W Br
coocH3
% SnBu3
H2/Pd(C) EtOA I
C
coocH3
N3
IV-4
0
o
LiOH/THF
1. (C1CO)Z/DCM
H20 9
%
COOCH3
COOH 2_ HZN
IV-5
\
N
IV-6
O
/ o
H
o
H
I
|
N
N
LiA1H4/1‘HF
W
\
|
IV-?
I
/
N
A method for preparing compounds found in table 4 is 55 protecting group is removed With a strong acid to afford the outlined in Scheme V. Substituted benZene compound such deprotected piperaZine. The free nitrogen is converted to the as 4-benZy1oXy-benZene (V-l) is reacted With sulfuryl chlo-
_
_
_
_
_
_
_
ride at 80o_120o C‘ to give a sulfonyl Chloride_ The sulfonyl
am1d1ne With a guanylatmg reagent such as amrdinosulfomc
chloride (V-Z) is reacted With Boc-piperaZine and the Boc
acid.
5,872,138 SCHEME V
0
100° 0/2 h >
O
V-1
EtgN/CHgClZ
>
v-2 0
o
NH
A general method for synthesizing compounds found in table 5 is outlined in Scheme VI. The ketone of a cyano benZophenone such as 1 is reduced With a metal hydride reagent such as sodium borohydride and the resultant alco
-Continued 40
SCHEME VI
hol is removed by treatment With dimethyldichlorosilane to
afford the diphenylmethane VI-3. The methoXy group is
1%
removed With borontribromide and the phenol is alkylated With an alkylating reagent such as benZyl bromide. The cyano group is then treated With sodium heXamethyldisila Zane and the silylimidate is hydrolyzed With an acidic
VI-3
Workup to afford the ?nal product. SCHEME VI 0 HO
NaBH4
%
CN
VI-4 55
H3CO
CN VI-1 H
OH
m (Me)2SiC12 H3CO
CN VI-2
6O
cszcmé
5,872,138 15 -continued
-continued
SCHEME VI
0
THF/CBr4
(C5H5)3P E NaN TMS 2 B 10
VI-5
CHZOH 1-4
15
20
0
CH3CN
4-AP >
25 . HCl
Br
VI-6
H2
1-5
NH 30
Unless otherwise shown, NMR data for the compounds identi?ed below was obtained using a ?eld strength of 400 MHZ.
Preparation of Compounds Found in Table I
; Bre
35
0
EXAMPLE 1
Preparation of 4-amino-1-(4-(1,1-diphenylmethoxy) benZylpyridinium bromide (1-6) @N
OH
NH;
\ /
CSZCO3/DMF 45 1-6
COZCHZCH3
Q CHBr
Step A:
Preparation of ethyl 4-(1,1-diphenylmethoxy)benZoate 50 1-1
1-2
55
0
Under N2, a mixture of ethyl 4-hydroxybenZoic acid (2.5 g, 15 mmol) 1-1, and CsZCO3 (5.0 g, 15 mmol) in DMF (50
ml) was stirred at room temperature. After 15 min, diphe nylmethyl bromide 1-2 (4.0 g, 16 mmol) was added and then heated at 60° C. After 1 h, saturated NaZCO3 was added and the mixture extracted with EtOAc (3x). The combined extracts were washed with H2O, dried, ?ltered and concen
::
trated to dryness to yield 1-3.
1H NMR (CDCl3) 6 1.3 (3H, t), 4.3 (2H, q), 6.95 (2H, s), 7.3
(10H, m) and 7.95 (2H, d). 60
Step B:
Preparation of 4-(1,1-diphenylmethoxy)benZyl alcohol
LAH; THF
(1-4) To a suspension of LAH (3.2 g, 0.084 mmol) in THF (400 ml) under N2 was added dropwise a solution of 1-3 (21 g,
COZCHZCH3 1-3
(1-3)
65
0.063 mol) in THF (100 ml). After stirring overnight at room temperature, saturated NaZSO4 was added to the reaction until a white suspension was observed. The mixture was
5,872,138 17
18
?ltered and the ?ltrate Was combined With H20 and CHCl3,
EXAMPLE 4
separated, and extracted further With CHCl3 (2>
5-2
HO
40
COZEt
100%
5'3
2-2 Was prepared as described for the preparation of 1-6
except 2-phenyl-1-bromoethane (2-1) Was used in place of 1-2 in Step A and synthesiZed. mp: 170°—1° C. (CH3CN);
1H NMR (dG-DMSO) 6 3.0 (2H, t), 4.2 (2H, t), 5.3 (2H, s), 6.82 (2H, d), 6.96 (2H, d), 7.3 (7H, m), 8.15 (2H, exch bs) and 8.27 (2H, d). Analysis calculated for C2OH21BrN2O C, 59.96; H, 5.71; N, 6.99
45
o
COZEt
50
Found: C, 59.62; H, 5.22; N, 7.14
5-4
52%
EXAMPLE 3
Preparation of 4-amino-1-(4-(cyclohexylmethoxy) benZyl)pyridinium bromide (3-3)
Step A: 55
3-3 Was prepared as described for the synthesis of 1-6
except 4-hydroxy-benZyl alcohol (3-1) and cyclohexylm ethyl bromide (3-2) Were used in Step A to yield
4-(cyclohexylmethoxy)benZyl alcohol directly. mp: 241°—3°
Preparation of 1,1-dicyclohexyl-2-hydroxy ethane (5-2) A solution of 1,1-dicyclohexylacetic acid (5-1) (2.8 g,
60
12.5 mmol) in THF (5 ml) under N2 Was cooled in an ice bath and then 1.0M borane in THF (17 ml, 17 mmol) Was added dropWise. After addition, the solution Was stirred at room temperature and then a 1:1 mixture of THF/H2O (10 ml) Was added carefully. The mixture Was added to saturated
1H NMR (dG-DMSO) 6 1.2 (5H, m), 1.75 (6H, m), 3.8 (2H, d), 5.28 (s, 2H), 6.8 (2H, d), 6.95 (2H, d), 7.34 (2H, d), 8.1 (2H, exch bs), 8.25 (2H, d).
Na2CO3 and extracted With EtOAc (3> 40
12 mmol) Was added. This mixture Was stirred overnight at ambient temperature and the solvent Was removed in vacuo. The residue Was taken up in the Water and the ether layer Was
separated, Washed With sodium bicarbonate and Water, dried, ?ltered and concentrated in vacuo to yield an oil. Chromato graphic puri?cation on silica gel gave 6-4 as a colorless oil.
6-1
45
1H NMR (d?-DMSO) 6 1.25 (3H, t), 2.1 (1H m), 2.25 (1H, m), 2.66 (1H, m), 2.75 (1H, m), 4.22 (2H, q), 5.4 (1H, t), 6.99 (2H, d), 7.12_7.44 (10H, m), 7.8 (2H, 01).
OH
50
Step C:
6-2
Preparation of 4-(1,3-diphenyl-1-propoxy)benZyl alcohol 55
(6-5)
[3-Phenylpropiophenone (6-1) (7.5 g, 36.0 mmol) Was suspended in ethanol (100 ml) and sodium borohydride (0.68 g, 18.0 mmol) Was added under N2. The resulting solution Was stirred at ambient temperature overnight. The ethanol Was removed in vacuo and the residual oil-solid Was 60
taken up in ethyl acetate (100 ml) and Water (30 ml). The ethyl acetate layer Was removed, Washed With Water, dried, ?ltered and concentrated in vacuo to give 6-2 as a colorless
oil.
1H NMR (d?-DMSO) 6 1.88 (2H, q), 2.6 (2H, m), 4.51 (1H,
q), 5.28 (1H, d), 7.1_7.37 (10H, m).
0 65
COOC2H5
5,872,138 21
22 Step E:
-continued
Preparation of 4-amino-1-[4-(1,3-diphenyl-1-propoxy) benZyl]pyridinium bromide (6-8)
10
CH3CN
NH;
OH
O
/ Br
I 6-7
\
Asolution of 6-4 (2.46 g, 6.8 mmol) in ether (30 ml) Was added dropWise under nitrogen atmosphere to a stirred cold suspension of lithium aluminum hydride (0.285 g, 7.5 mmol) in ether (5 ml). The mixture Was stirred for 3 hrs at ambient temperature and Was decomposed by carefully
6-6
15
20
Q} e9Bre
adding Water and sodium hydroxide. This mixture Was extracted With ethyl acetate. The extract Was Washed With Water, dried, ?ltered and concentrated in vacuo to yield 6-5
N
as a colorless viscious oil.
1H NMR (d?-DMSO) 6 2.05 (1H, m), 2.2 (1H, m), 2.66 (1H,
N
25
6-8
|
NHZ
m), 2.75 (1H, m), 4.34 (2H, d), 4.99 (1H, t), 5.26 (1H, t), 6.81 (2H, 01), 7044.4 (12H, m).
To a solution of 4-(1,3-diphenyl-1-propoxy)benZyl bro
Step D:
Preparation of 4-(1,3-diphenyl-1-propoxy)benZyl bro mide (6-6)
30
mide 6-6 (0.381 g, 1.0 mmol) in acetonitrile (7 ml) Was
added 4-aminopyridine (6-7) (94 mg, 1.0 mmol) With stir ring at room temperature. The mixture Was stirred for 3 hrs
35
40
and the resulting White suspension Was ?ltered. The White solid obtained 6-8, melted at 215°—216.5° C. Recrystalliza tion from acetonitrile raised the melting point to 218.5°—220° C.
1H NMR (d?-DMSO) 6 2.06 (1H, m), 2.2 (1H, m), 2.65 (1H, m), 2.75 (1H, m), 5.2 (2H, s), 5.3 (1H, m), 6.8 (2H, d), 6.9 (2H, d), 7.16—7.41 (12H, m), 8.12 (2H, s), 8.24 (2H, 01). Analysis calculated for C27H27N2OBr C, 68.21; H, 5.73; N, 5.89
Found: C, 68.48; H, 5.68; N, 5.98 45
EXAMPLE 7
Preparation of 4-amino-1-[4-(2,2-diphenylethoxy) benZyl]pyridinium bromide (7-7) 50
Step A:
Preparation of ethyl 4-(2,2-diphenylethoxy)benZoate
(7-3) 55
To an ice cold solution of 6-5 (1.19 g, 6.0 mmol) in
acetonitrile (15 ml) Was added pyridine (0.76 g, 9.6 mmol) and then triphenylphosphine dibromide (3.29 g, 7.8 mmol). The mixture Was stirred at ice bath temperature for 1A1 hr
under N2 and at ambient temperature for 1 &% hrs and then
60
Was ?ltered. Concentration of the ?ltrate in vacuo gave a
CH—CH2OH éOH DEAD 7-1
crude oil. The pure product 6-6 Was obtained after three
silica gel chromatographic separations. 1H NMR (d?-DMSO) 6 2.06 (1H, m), 2.2 (1H, m), 2.66 (1H, m), 2.75 (1H, m), 4.62 (2H, s), 5.3 (1H, t), 6.82 (2H, d), 7.16—7.4 (12H, m).
65
COOC2H5
5,872,138 23
24
-continued
(0.204 g, 0.75 mmol) Was added. After 1/2 hr. the reaction Was
diluted With methanol (2 ml) and Water (5 ml). The ether layer Was separated, Washed With sodium bicarbonate and Water, dried and ?ltered. Concentration of the ?ltrate in vacuo gave 7-5 as a White solid. mp: 70°—73° C. CH—CH2— O
1H NMR (dG-DMSO): 6 4.43—4.62 (3H, m), 4.64 (2H, s), 6.9 Step D: (2H, d), 7.15—7.4 (12H,
COOC2H5
7-3
10
Preparation of 4-amino-1-[4-(2,2-diphenyl-ethoxy) benZyl]pyridinium bromide (7-7)
2,2-Diphenylethanol (7-1) (5.95 g, 30 mmol) and ethyl 4-hydroxybenZoate (7-2) (5.48 g, 30 mmol) Were reacted, as in Example 6. 7-3 Was obtained after chromatography as a
CH—CH2—O
colorless oil.
1H NMR (CDCl3): 6 1.36 (3H, t), 4.33 (2H, q), 4.52 (3H, s), 6.9 (2H, d), 7.2—7.36 (10H, m), 7.96 (2H, d).
W Br
7-5
Step B:
/
Preparation of ethyl 4-(2,2-diphenylethoxy)benZyl alco hol (7-4)
I
\
N 7-6
LAH N2
EtZO E
25
CH-CHg-O
BIG
7-3 Ne19 _
7-7 30
43/
OH 7-4
7-5 (100 mg, 0.27 mmol) Was reacted With 35
7-3 (0.757 g, 2.2 mmol) Was dissolved in ether (20 ml)
ml) as in Example 6, Step E. The White solid product 7-7 Was
and Was reduced With lithium aluminum hydride (92 mg, 2.4 mmol) under nitrogen as in Example 6, Step B. 7-4 Was
obtained. mp: 137°—139° C.
obtained as a colorless oil.
1H NMR (dG-DMSO) 6 1.55 (1H, t), 4.45—4.52 (3H, m), 4.61
4-aminopyridine 7-6 (28 mg, 0.30 mmol) in acetonitrile (3
40
(2H, d), 6.88 (2H, d), 7.2—7.35 (12H, Step C:
Preparation of 4-(2,2-diphenylethoxy)benZyl bromide
(7-5)
1H NMR (dG-DMSO) 6: 4.45—4.59 (3H, m), 5.27 (2H, s), 6.82 (2H, d), 6.98 (2H, d), 7.17—7.4 (12H, m), 8.13 (2H, s), 8.27 (2H, d). Analysis calculated for (C26H25N2OBr.0.5 H2O): C, 66.38; H, 5.57; N, 5.96 Found: C, 66.57; H, 5.42; N, 5.98
45
EXAMPLE 8
Preparation of 4-amino-1-[4-(dicyclohexylmethoxy) benZyl]pyridinium bromide (8-7)
PBT3
EtZOE
50
OH
Step A:
Preparation of ethyl 4-(dicyclohexylmethoxy)benZoate
(8-3) 55
65
7-4 (0.572 g, 1.9 mmol) Was dissolved in ether (10 ml). The solution Was cooled in ice and phosphorus tribromide
5,872,138 25
26
-continued
-continued
CH—O
COOC2H5 Br
8-3 10
Dicyclohexylmethanol (8-1) (4.91 g, 25 mmol) and ethyl 8-4 (1.93 g, 6.4 mmol) Was converted to the bromide
4-hydroxybenZoate (8-2) (4.15 g, 25 mmol) Were reacted, as in Example 6, Step B, to obtain a colorless oil 8-3 after
15
chromatography. 1H NMR (CDCl3): 6 0.98—1.3 (11H, m), 1.36 (3H, t), 1.58—1.86 (11H, m), 4.02 (1H, t), 4.33 (2H, q), 6.92 (2H, d), 7.94 (2H, 01).
using phosphorus tribromide (1.69 g, 2.6 mmol) in ether (25 ml) as in Example 7. Recovered 8-5 liquid.
1H NMR (CDC13) 6 1.0_1.32 (11H, m), 1.53—1.84 (11H, m), 3.91 (1H, t), 4.49 (2H, s), 6.85 (2H, d), 7.25 (2H, 01).
Step B:
Preparation of 4-(dicyclohexylmethoxy)benZyl alcohol Step D:
(8-4) 25
Preparation of 4-amino-1-[4-(dicyclohexylmethoxy) benZyl]pyridinium bromide (8-7)
LAH N2
CH—O
COOCZHS W
83
35
W 4}. 8-5
CH3CN
NH; 5
/ I \
N 8-6 OH
45
Reduction of 8-3 (2.8 g, 8.1 mmol) With lithium alumi num hydride (0.338 g, 8.9 mmol) in ether (30 ml) as in
/
I 8-7
Example 6 gave 8-4 as a colorless oil.
1H NMR (CDC13): 6 1.01_1.31 (11H, m), 1.45—1.86 (11H,
NHZ
m), 3.91 (1H, t), 4.6 (2H, d), 6.9 (2H, d), 7.23 (2H, 01). Step C:
Preparation of 4-(dicyclohexylmethoxy)benZyl bromide
(8-5)
8-5 (183 mg, 0.5 mmol) and 4-aminopyridine 8-6 (47 mg, 55
0.5 mmol) Were reacted in acetonitrile (4 ml) as in Example 1. Recovered White solid 8-7. mp: 248°—249° C.
1H NMR (d?-DMSO) 6 0.94—1.28 (11H, m), 1.5_1.75 (11H, m), 4.1 (1H, t), 5.24 (2H, s), 6.83 (2H, d), 6.98 (H, d), 7.28 (2H, d), 8.14 (2H, s), 8.3 (2H, 01).
PBT3
CH—O
who OH
Analysis calculated for C25H35N2OBr C, 65.35; H, 7.68; N,
8-4
6.10 65
Found: C, 65.45; H, 7.62; N, 6.17
5,872,138 27
28
EXAMPLE 9
-continued
Preparation of 3,4-diamino-1-(4-benZyloxybenZyl) pyridinium bromide (9-7) Step A:
5
0
Preparation of 4-benZyloxybenZyl bromide (9-2)
Ne; BIG /
9-4 PBr3
@—\ 0
I
\
EtZO
1O
NH;
NH2
OH 9-1
15
9-2 (1.38 g, 5.0 mmol) Was reacted With 3,4
diaminopyridine (9-3) (0.573 g, 5.25 mmol) in acetonitrile O
(50 ml) as in Example 1. Recovered White solid 9-4. RecrystalliZed from acetonitrile, mp: 240°—241° C.
Br 9-2
20 1H NMR (dG-DMSO) 6 5.1 (2H, s), 5.25 (2H, s), 5.56 (2H,
To a cold suspension of 4-benZyloxybenZyl alcohol 9-1
3’
(21.4 g, 0.10 mol) in ether (250 ml) Was added phosphorus
’
(31))’ 7'05 (2H’ d)’ 7'24_7'46 (9H’ m)’ 7'61 (1H’ '
’
'
tribromide (10.8 g, 40 mmol) as in Example 7. Recovered 27.2 g of White solid Which Was recrystalliZed from hexane 25 Analysis calculated for (C19H2ON3O+Br_): (0.25 EtZO) C,
(200 ml). Obtained pure bromide 9-2.
59.09; H, 5.22; N, 10.88
1H NMR (CDCl3) 6 4.5 (2H, s), 5.05 (2H, s), 6.94 (2H, d), 7'26_7'45 (7H’ Found: C, 59.33; H, 5.60; N, 10.38 Step B: Preparation of 3,4-diamino-1-(4-benZyloxybenZyl)- 30 pyridinium bromide (9-4)
The compounds shoWn in the table beloW are exemplary compounds of the present invention. The range of Ki values
associated With the speci?cally listed compounds is repre CH3CN 0
E
N
sented as follows: 35
/
Br
\
9Q
I
9'3 NHZ
+
0.1 ,uM and 1.0 ,uM
NH;
TABLE I R1 _
R
\
Xe
/
@
Z—Y
N
\
/
R2
NH; Ki
(thr) R
R1
Z
Y
R2
,uM mp
H
CH CH H
++
H
CH CH H
+++
CHZO —
5,872,138 TABLE I-continued R1 Xe
R
\
/
@
Z—Y
N
\
/
R2
NHZ Ki
(thr) R
R1
Z
Y
R2
CH CH H
CH2O—
,uM mp +++ 125-7O
—OCH2
H
CH CH NH2
++
240-1O
CH2O—
The general procedure outlined in Scheme II can be used
Was concentrated to dryness and ?ushed once With CHCl3.
to make the following compounds.
The residue Was dissolved in CHCl3 (60 ml) and added dropWise to a suspension of 4-AP (6.2 g, 66 mmol) in CHCl3
EXAMPLE 10
30
Preparation of N-4-(4-benZyloxybenZyl)
(100 ml). After 1 h at room temperature the solution Was
poured into saturated NaZCO3 and separated. The aqueous Was further extracted With EtOAc (3>
1H NMR (CDC13) 6 0.91 (3H, d), 1.05 (3H, d), 1.32 (3H, t), 2.15 (1H, m), 4.3 (2H, q), 4.88 (1H, d), 6.83 (2H, 01), 7.24.4
(5H, m), 8.7 (2H, 01). 13-1
13-2
Step C: Isobutyrophenone 13-1 (29.6 g, 0.20 mmol) Was dis solved in ethanol (300 ml) and Was reduced under N2 With sodium borohydride (7.57 g, 0.20 mmol) for 4 hrs. at room
35
Preparation of 4-(2-methyl-1-phenylpropoxy)benZoic acid (13-6)
temperature. The ethanol Was removed in vacuo and the
residue Was taken up in ethyl acetate (200 ml) and Water (100 ml). The ethyl acetate extract Was Washed With Water,
40
dried, ?ltered and concentrated in vacuo to obtain alcohol 13-2.
KOH
0 Q cooczn5 EtOH/H2O
1H NMR (CDCl3): 0 0.82 (3H, d), 1.0 (3H, d), 1.83 (1H, s), 2.0 (1H, m), 4.4 (1H, d), 7.26 (5H,
13-5 45
Step B:
Preparation of ethyl 4-(2-methyl-1-phenylpropoxy) benZoate (13-5)
O
COOH
OH
NaH Cl3CCN
50
E N2
13-2
55 NH
To 13-5 (8.89 g, 29.8 mmol) Was added 2N potassium
hydroxide (100 ml) and ethanol (50 ml). The mixture Was
O)\cc13
stirred at 100° C. overnight. The ethanol Was removed in
cyclohex. CHZClZ
OH
vacuo and the aqueous solution Was cooled in ice and
E 60
13-3
acidi?ed With 6N HCl (35 ml, 0.21 mol). The acid Was extracted into ethyl acetate (50 ml) and the extract Was Washed With Water, dried, ?ltered and concentrated in vacuo to obtain the oil 13-6, Which solidi?ed. The White solid melted at 125°—129° C.
134
cooczns
65
1H NMR (CDC13) 6 0.91 (3H, d), 1.05 (3H, d), 2.15 (1H, m), 4.9 (1H, d), 6.85 (2H, 01), 724.35 (5H, m), 7.91 (2H, d).
5,872,138 35
36
Step D:
Step F:
Preparation of 4-(2-methyl-1-phenylpropoxy)benZoyl chloride (13-7)
benZylamino]pyridine (13-10)
Preparation of 4-[4-(2-methyl-1-phenylpropoxy) 0
0
oxal l chloride
0 Q COOH
. L1AlH4
N2
/ /H W N
CHZCIZ 13-9
/
I
13-6 N
15
13-10
I
20
13-6 (0.541 g, 2.0 mmol) Was stirred in methylene chlo ride (3 ml) at room temperature and oxalyl chloride (0.381
To a stirred suspension of lithium aluminum hydride
g, 3.0 mmol) Was added. After 3 hrs. at room temperature the solution Was concentrated in vacuo to a pale yelloW liquid 13-7. The acid chloride Was used Without puri?cation.
25
Step E:
30
ml) over a 5 min period. The mixture Was stirred at ambient
temperature overnight and then Was decomposed by adding Water and sodium hydroxide. Ethyl acetate (25 ml) Was
Preparation of 4-(2-methyl-1-phenylpropoxy)-N-(4
pyridyl)benZamide (13-9) 35 0
||
0
Et3N THF
c—c1
W
I
\
13-7
13-8
40
/
/o /
added and the mixture Was ?ltered. The ?ltrate Was dried, ?ltered and concentrated in vacuo to 620 mg of colorless oil Which Was chromatographed on silica gel. 13-10 Was obtained as an oil. The hydrochloride salt melted at
186°—188° C.
1H NMR (dG-DMSO) 0 0.5 (3H, d), 0.99 (3H, d), 1.16 (1H, t), 4.15 (2H, d), 4.99 (1H, d), 6.47 (2H, d), 6.82 (2H, d), 7.05 (1H, t), 7.12 (2 H, d), 7.2—7.25 (1H, m), 7.25—7.36 (4H, m), 7.96 (2H, d). Analysis calculated for (C22H24N2O.HCl.0.2 H2O) C, 70.93; H, 6.87; N, 7.52 Found: C, 70.99; H, 6.79; N, 7.62
N
O
(0.152 g, 4.0 mmol) in ether (5 ml) under nitrogen Was added a solution of 13-9 (0.71 g, 2.0 mmol) in tetrahydrofuran (5
H
45
EXAMPLE 14
N
13-9
I
\ /
N
Preparation of 4-[4-(2,2-diphenylethoxy)
benZylamino]pyridine (14-7) 50
Step A:
Preparation of ethyl 4-(2,2-diphenylethoxy)benZoate (14
3) To an ice cold stirred solution of 13-8 (0.188 g, 2.0 mmol)
and triethylamine (0.233 g, 2.0 mmol) in tetrahydrofuran (10
55
ml) Was added a solution 13-7 (0.54 g, 2.0 mmol) over a 5-10 min period. The reaction Was stirred at ambient tem perature overnight. Then the THF Was removed in vacuo and
Ph3P N; THE
the residue Was taken up in ethyl acetate (50 ml) and Water (25 ml). The ethyl acetate extract Was separated, Washed With Water, dried and concentrated in vacuo to yield the oil
CH—CH2OH éOH DEAD
U
13-9.
1H NMR (d?-DMSO) 6 0.84 (3H, d), 1.02 (3H, d), 2.12 (1H, m), 5.2 (1H, d), 7.0 (2H, d), 7.2_7.4 (5H, m), 7.72 (2H, d), 7.82 (2H, d), 8.42 (2H, d), 10.33 (1H, s).
65
14-1 14 2
5,872,138 37
38
-Continued
DMF (5 ml) under nitrogen Was added 1-ethyl-3-(3
dimethylamino-propyl)carbodiimide hydrochloride (113 mg, 0.59 mmol). The mixture Was stirred at ambient tem
perature overnight and the DMF Was removed under high 5 vacuum. The residual oil Was taken up in ethyl acetate (15
CH_CHZ_O
COOCZHS
ml) and Washed With Water and saturated sodium chloride,
14_3
dried, ?ltered and concentrated to an amber gum 14-6.
10
Step D: Preparation of 4-[4-(2,2-diphenylethoXy)benZylamino]
pyridine (14-7) To 2,2-diphenylethanol (14-1) (5.95 g, 30 mmol) Was
added ethyl 4-hydroXybenZoate (14-2) (5.40 g, 30 mmol), triphenylphosphine (8.66 g, 33 mmol) and tetrahydrofuran 15 (125 ml). The miXture Was cooled in ice and diethyl aZodi-
O
carboXylate (5.75 g, 33 mmol) Was added. The reaction
CH-CHg-O
miXture Was stirred under nitrogen at ambient temperature overnight and then the volatile components Were removed in vacuo. The oil-solid residue Was taken up in ethyl acetate 20
LAH N2
/ /H W N
14'6 /
(100 ml) and Water (50 ml). Work up of the ethyl acetate
I
portion give crude solid. The product 14-3 Was puri?ed by silica gel chromatography.
\
N
1H NMR (CDCl3) 6 1.37 (3H, t), 4.33 (2H, q), 4.52 (3H, s),
6.9 (2H, Step B:
d), 7.4—7.66 (10H, m), 7.96 (2H, d).
25
Q
Preparation of 4-(2,2-diphenylethoXy)benZoic acid (14-4) NaOH
CH—CH2—O—©—COOC2H5W
CH_CH2_O 4®_\ /H
Q
30
14_7
N
/
14-3
\
I N
CH_CHZ_O—©_COOH 35
@/
144
To a solution of 14-6 (0.144 g, 0.365 mmol) in tetrahy
drofuran (1 ml) and ether (3 ml) under nitrogen Was added lithium aluminum hydride (30 mg, 0.79 mmol). The mixture
A
_ sus
ens1ono
£14 3646 -
10 m
,
.
1)_ 1017 mmo
in
Was stirred at room temperature overnight and then Was
0a
ueous
~
~
~
~
Sodium ‘iydmxide (15 m1) andithanol (5 ml) was aired at 40 ?fggggjggdagggggggjgjgjgggugygrjfjjgggg3131f; 100° C. overnight. The resulting solution Was cooled and
acidi?ed With eXcess HCl. A gum separated Which Was eXtracted into ethyl acetate (15 ml), Washed With Water, dried, ?ltered and concentrated in vacuo. The acid 14-4 Was
-
-
- -
Whlch Was pun?ed by slhca gel chromftography and Con Verted to the Hcl Salt’ mp: 213 _2145 0 1H NMR (d6-DMSO) 5 3.36 (1H, S), 4.36—4.58 (6H, d, m),
a pale yelloW solid, mp: 140°—146° C. 45 6.84—7.0 (3H, d, bm), 7.16—7.4 (11H, m), 8.06—8.25 (2H, d), 1H NMR (CDCl3) 6 4.55 (3H, s), 6.94 (2H, d), 7.21—7.37 9.08—9.24 (1H, bs). 8'04 (2H’ (1) Analysis calculated for (C22H24N2O.HCl) C, 74.48; H, 6.10; Preparation of 4-(2,2-diphenylethoXy)-N-(4-pyridyl) N’ 6'74 benZamide (14-6) Found: C, 74.89; H, 6.04; N, 6.72
O
CH-CHg-O @ COOH HOBTNH; EDC TEA DMF 14-4
/
N
|
\
CH-CHg-O
/
14-6 Z
N
/H N
/ \
14-5
To 14-4 (125 mg, 0.393 mmol), 4-aminopyridine (14-5)
(56 mg, 0.59 mmol), 1-hydroXybenZotriaZole hydrate (80 mg, 0.59 mmol) and triethylamine (60 mg, 0.59 mmol) in
N
|
