CA2303832A1 - Thrombin inhibitors - Google Patents
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- CA2303832A1 CA2303832A1 CA002303832A CA2303832A CA2303832A1 CA 2303832 A1 CA2303832 A1 CA 2303832A1 CA 002303832 A CA002303832 A CA 002303832A CA 2303832 A CA2303832 A CA 2303832A CA 2303832 A1 CA2303832 A1 CA 2303832A1
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- C07—ORGANIC CHEMISTRY
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- C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
- C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
- C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
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- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
- A61P7/02—Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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Abstract
A compound which inhibits human thrombin and which has general structure (I), such as (II).
Description
TITL E OF TI-IE INVENTTf~N
THROMBIN INHIBITORS
BACKGROLTIv_TD OF THE INVFNmTnN
Thrombin is a serine protease present in blood plasma in the form of a precursor, prothrombin. Thrombin plays a central role in the mechanism of blood coagulation by converting the solution plasma protein, fibrinogen, into insoluble fibrin.
Edwards et al., J. Amer. Chem. Soc., (1992) vol. 114, pp.
1854-63, describes peptidyl a-ketobenzoxazoles which are reversible inhibitors of the serine professes human leukocyte elastase and porcine pancreatic elastase.
European Publication 363 284 describes analogs of peptidase 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 8624567? also describes peptidase inhibitors having an activated electrophilic ketone moiety such as fluoromethylene ketone or a-keto carboxyl derivatives.
Thrombin inhibitors described in prior publications contain 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 liver toxicity.
European Publication 601 459 describes sulfonamido heterocyclic thrombin inhibitors, such as N-[4-[(aminoimino-methyl)amino)butyl]-1-[N-(2-naphthalenylsulfonyl)-L-phenylalanyl]-L-prolinamide.
WO 94/29336 describes compounds which are useful as thrombin inhibitors.
SUMMARY OF THE NVFNTT N
A compound which inhibits human thrombin and which has the general structure H O R
i R2'N N N~H
Rt H n N
such as H O
N\~
SO~ _ N NH
N
The invention also includes a composition for preventing or treating unstable angina, refractory angina, myocardial infarction, transient ischemic attacks, atrial fibrillation, thrombotic stroke, embolic stroke, deep vein thrombosis, disseminated intravascular coagulation, ocular build up of fibrin, and reocclusion or restenosis of recanalized vessels, 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 invention includes a composition for inhibiting loss of blood platelets, inhibiting formation of blood platelet aggregates, inhibiting formation of fibrin, inhibiting thrombus 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 blood, blood products, or mammalian organs in order to effect the desired inhibitions.
The invention also includes the use of a compound of the invention in the manufacture of a medicament for preventing or treating unstable angina, refractory angina, myocardial infarction, transient ischemic attacks, atrial fibrillation, thrombotic stroke, embolic stroke, deep vein thrombosis, disseminated intravascular coagulation, ocular build up of fibrin, 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.
DFTATT FD DESCRIPTTO1V OF TH INVENTTC~rr Compounds of the invention have the following structure:
H O R
RAN N N~H
R~ H
N
and pharmaceutically acceptable salts thereof, wherein nis0orl;
R is hydrogen C 1_g alkyl, arylCO_galkylene-, C3_gcycloalkyl, or -CH2Cg_gcycloalkyl;
R1 is hydrogen, C1_galkyl hydroxy, C1_6 alkyl, -(CH2)mS02R3, wherein m is 0-6, arylCO_galkylene, wherein aryl is unsubstituted or mono-, di-, tri-, or tetra- substituted with C1_4 alkyl, hydroxy, C1_4alkoxy, or CF3;
-(CH2)mSR3, wherein m is 0-6, or C3_gcycloalkyl;
R2 is Boc, -S02R4, -S02-CH=CHR4, hydrogen, C 1_6alkyl, C3_gcycloalkyl, R4, -C(O)R5, or -S02R5;
R3 is C1_galkyl, or R4 is arylCO_salkylene, wherein aryl is unsubstituted or mono-, di-, tri-, or tetra- substituted with C1_4 alkyl, hydroxy, C 1-4~oxy, halogen, amino, or CF3;
R5 is 30 hydrogen, C 1_galkyl, C3_gcycloalkyl, or arylCO_galkyi.
THROMBIN INHIBITORS
BACKGROLTIv_TD OF THE INVFNmTnN
Thrombin is a serine protease present in blood plasma in the form of a precursor, prothrombin. Thrombin plays a central role in the mechanism of blood coagulation by converting the solution plasma protein, fibrinogen, into insoluble fibrin.
Edwards et al., J. Amer. Chem. Soc., (1992) vol. 114, pp.
1854-63, describes peptidyl a-ketobenzoxazoles which are reversible inhibitors of the serine professes human leukocyte elastase and porcine pancreatic elastase.
European Publication 363 284 describes analogs of peptidase 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 8624567? also describes peptidase inhibitors having an activated electrophilic ketone moiety such as fluoromethylene ketone or a-keto carboxyl derivatives.
Thrombin inhibitors described in prior publications contain 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 liver toxicity.
European Publication 601 459 describes sulfonamido heterocyclic thrombin inhibitors, such as N-[4-[(aminoimino-methyl)amino)butyl]-1-[N-(2-naphthalenylsulfonyl)-L-phenylalanyl]-L-prolinamide.
WO 94/29336 describes compounds which are useful as thrombin inhibitors.
SUMMARY OF THE NVFNTT N
A compound which inhibits human thrombin and which has the general structure H O R
i R2'N N N~H
Rt H n N
such as H O
N\~
SO~ _ N NH
N
The invention also includes a composition for preventing or treating unstable angina, refractory angina, myocardial infarction, transient ischemic attacks, atrial fibrillation, thrombotic stroke, embolic stroke, deep vein thrombosis, disseminated intravascular coagulation, ocular build up of fibrin, and reocclusion or restenosis of recanalized vessels, 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 invention includes a composition for inhibiting loss of blood platelets, inhibiting formation of blood platelet aggregates, inhibiting formation of fibrin, inhibiting thrombus 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 blood, blood products, or mammalian organs in order to effect the desired inhibitions.
The invention also includes the use of a compound of the invention in the manufacture of a medicament for preventing or treating unstable angina, refractory angina, myocardial infarction, transient ischemic attacks, atrial fibrillation, thrombotic stroke, embolic stroke, deep vein thrombosis, disseminated intravascular coagulation, ocular build up of fibrin, 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.
DFTATT FD DESCRIPTTO1V OF TH INVENTTC~rr Compounds of the invention have the following structure:
H O R
RAN N N~H
R~ H
N
and pharmaceutically acceptable salts thereof, wherein nis0orl;
R is hydrogen C 1_g alkyl, arylCO_galkylene-, C3_gcycloalkyl, or -CH2Cg_gcycloalkyl;
R1 is hydrogen, C1_galkyl hydroxy, C1_6 alkyl, -(CH2)mS02R3, wherein m is 0-6, arylCO_galkylene, wherein aryl is unsubstituted or mono-, di-, tri-, or tetra- substituted with C1_4 alkyl, hydroxy, C1_4alkoxy, or CF3;
-(CH2)mSR3, wherein m is 0-6, or C3_gcycloalkyl;
R2 is Boc, -S02R4, -S02-CH=CHR4, hydrogen, C 1_6alkyl, C3_gcycloalkyl, R4, -C(O)R5, or -S02R5;
R3 is C1_galkyl, or R4 is arylCO_salkylene, wherein aryl is unsubstituted or mono-, di-, tri-, or tetra- substituted with C1_4 alkyl, hydroxy, C 1-4~oxy, halogen, amino, or CF3;
R5 is 30 hydrogen, C 1_galkyl, C3_gcycloalkyl, or arylCO_galkyi.
WO 99116750 PCTIUS98I20t74 In a class of compounds and pharmaceutically acceptable salts thereof, of the invention, the compounds have the structure H O R
R2~N N N~H
R~ H
N
and R is -CH ~ ~ -CH2 or -CH(CH3)2.
In a subclass of this class of compounds and pharmaceutically acceptable salts thereof, R1 is hydrogen, -CH20H, CH3, -CH2CH2SCH3, -CH2CH2S02CHg.
In a group of this subclass of compounds and pharmaceutically acceptable salts thereof, R2 is hydrogen, O C H3 _ ~O~CH3 ~-S02 CH3 , -S02 CH2 ~ / ~ ~-S02CH2 CI
/
~- S02 / ~ / - SO
/ cl H3C _ CH3 CI
-S02 ~ / OCH3 ~-S02 CI
/
-SO- HC=HC ~ ~ CI ~-S- HC=HC
2 , 02 ~ / , CI
-SO 2CH2 ~ / CF3 ~r ~-S02CH2 CI
~ /
R2~N N N~H
R~ H
N
and R is -CH ~ ~ -CH2 or -CH(CH3)2.
In a subclass of this class of compounds and pharmaceutically acceptable salts thereof, R1 is hydrogen, -CH20H, CH3, -CH2CH2SCH3, -CH2CH2S02CHg.
In a group of this subclass of compounds and pharmaceutically acceptable salts thereof, R2 is hydrogen, O C H3 _ ~O~CH3 ~-S02 CH3 , -S02 CH2 ~ / ~ ~-S02CH2 CI
/
~- S02 / ~ / - SO
/ cl H3C _ CH3 CI
-S02 ~ / OCH3 ~-S02 CI
/
-SO- HC=HC ~ ~ CI ~-S- HC=HC
2 , 02 ~ / , CI
-SO 2CH2 ~ / CF3 ~r ~-S02CH2 CI
~ /
iI
Specific exemplifications of this class are shown in the table below Table 1 H O R
R2~N N N~H
R~ H n N
R n R2 R1 Ki(~,M) Bn 0 Boc H
Bn 0 PhS02- H
c-hexylCH2- 0 PhS02- H
Bn 0 4-ClPhS02- H
Bn 1 4-Cl-PhS02- H **
Bn 1 B~2- H **
Bn 1 2-naphthS02- H **
Bn 1 3,4-di-Cl-PhS02-H **
Bn 1 PhCH=CHS02- H **
Bn 1 4-Cl-BnS02- H **
Bn 1 4-CF3-BnS02- H **
Bn 1 3,4-di-Cl-BnS02-H **
Bn 1 4-OCH3-2,3,6- H **
MePhS02-Bn 1 3,4-di-Cl-PhS02-(S)-CH20H **
* > 1.0 E.~M
**<_1.ONM
_7_ Tabl 1 ( ,nnt'rl) H O R
R2~N N N~H
R1 H n y N
R n R2 R1 ~(~) Bn 1 BnS02- (R)-CH3 Bn 1 BnS02- (S)-CHg **
Bn 1 BnS02- (S)-CH2CH2S02CH3 **
Bn 1 BnS02- (S)-CH2CH2SCHg **
Bn 1 H 4-CIBn- **
(CH3)2CH2- 1 BnS02- H
(CHg)2CH2- I 3,4-di-Cl-PhS02-H **
* > 1.0 N,M
** 5 L0 N,M
Exemplifications also include the pharmaceutically acceptable salts of the above-identified compounds.
_g_ ABBREVIATIONS
Desienation Protecting =ro m BOC (Boc) t-butyloxycarbonyl CBZ (Cbz) benzyloxycarbonyl(carbobenzoxy) Ph phenyl Bn benzyl TBS (TBDMS) t-butyl-dimethylsilyl Activating GrOUD
HBT(HOBT or HOBt) 1-hydroxybenzotriazole hydrate Designation ~Dli_n_~ Reagg~t BOP reagent benzotriazol-1-yloxytris-(dimethylamino)phosphonium hexafluorophosphate BOP-Cl bis(2-oxo-3-oxazolidinyl)phosphinic chloride EDC 1-ethyl-3-(3-dimethylaminopropyl ) carbodiimide hydrochloride _g_ Other (BOC~O (BOC20) di-t-butyl dicarbonate n-Bu4N+F- tetrabutyl ammonium fluoride nBuLi (n-Buli) n-butyllithium DMF dimethylformamide EtgN (TEA) triethylamine EtOAc ethyl acetate TFA trifluoroacetic acid DMAP dimethylaminopyridine DME dimethoxyethane NMM N-methylmorpholine DPPA diphenylphosphoryl azide THF tetrahydrofuran DPPC diphenylphosphinic chloride DIPEA diisopropylethylamine Amino Acid Ile Isoleucine Phe Phenylalanine Pro Proline Ala Alanine V al Valine 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. The compounds of the present invention may also have polymorphic crystalline forms, with all polymorphic crystalline forms being included in the present invention.
10 When any variable occurs more than one time in any constituent or in formula I, its definition on each occurrence is independent of its definition at every other occurrence. Also, combinations of substituents and/or variables are permissible only if such combinations result in stable compounds.
As used herein, except where noted, "alkyl" is intended to include both branched- and straight-chain saturated aliphatic hydrocarbon groups having, unless otherwise noted, 1-8 carbon atoms (Me is methyl, Et is ethyl, Pr is propyl, Bu is butyl). "Alkenyl" is 5 intended to include both branched- and straight-chain unsaturated aliphatic hydrocarbon groups having, unless otherwise noted, 1-8 carbon atoms, e.g. ethenyl, propenyl, etc. "Cycloalkyl" includes cyclic saturated aliphatic hydrocarbon groups having 3-8 carbon atoms (e.g. "C3_ gcycloalkyl" is intended to include cyclopropyl, cyclobutyl, cyclopentyl, 10 cyclohexyl and cycloheptyl, and the like). The term "C7-12 bicyclic alkyl"
is intended to include bicyclo[2.2.1]heptyl (norbornyl), bicyclo[2.2.2]octyl, 1,1,3-trimethyl-bicyclo[2.2.1]heptyl (bornyl), and the like. The term "aryl" as used herein except where noted, represents a stable 6- to 10-membered mono- or bicyclic ring system such as phenyl, or naphthyl:
15 The aryl ring can be unsubstituted or substituted with one or more of C1_ 4 lower alkyl; hydroxy; alkoxy; halogen; amino. The term "heteroaryl"
refers to a 5- to 7- membered unsaturated ring containing 1 or 2 heteroatoms selected from O, N, or S. "Alkoxy" represents an alkyl group having 1-8 carbon atoms attached through an oxygen bridge.
20 "Halo", as used herein, means fluoro, chloro, bromo and iodo.
"Counteriori' is used to represent a small, single negatively-charged species, such as chloride, bromide, hydroxide, acetate, trifluroacetate, perchlorate, nitrate, benzoate, maleate, tartrate, hemitartrate, benzene sulfonate, and the like.
25 Under standard nomenclature used throughout this disclosure unless specified otherwise, the terminal portion of the designated side chain is described first followed by the adjacent functionality toward the point of attachment. For example, an ethyl substituent substituted with "methylcarbonylamino" is equivalent to O
The term "heterocycle" or "heterocyclic ring", as used herein except where noted, represents a stable 5- to 7-membered mono-or bicyclic or stable 7- to 10-membered bicyclic 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 of N, O and S, and wherein the nitrogen and sulfur heteroatoms may optionally be oxidized, and the nitrogen heteroatom may optionally be quaternized, and including any bicyclic group in which any of the above-defined heterocyclic rings is fused to a benzene ring. Especially useful are rings containing one oxygen or sulfur, one to four nitrogen atoms, or one oxygen or sulfur combined with one or two nitrogen atoms. The heterocyclic ring may be attached at any heteroatom or carbon atom which results in the creation of a stable structure. Examples of such heterocyclic elements include piperidinyl, piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolodinyl, 2-oxoazepinyl, azepinyl, pyrrolyl, 4-piperidonyl, 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, thiamorpholinyl sulfone, and oxadiazolyl. Morpholino is the same as 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 salts or the quaternary ammonium salts which are formed, e.g., from inorganic or organic acids or bases.
Examples of such acid addition salts include acetate, adipate, alginate, 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, 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 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, and butyl chloride, bromides and iodides;
dialkyl sulfates like dimethyl, diethyl, dibutyl and diamyl sulfates, long chain halides such as decyl, Iauryl, myristyl and stearyl chlorides, bromides and iodides, aralkyl halides Iike benzyl and phenethyl bromides and others.
Thrombin Inhibitors - Thera;~pmt;r~. Tl~eP~
Anticoagulant therapy is indicated for the treatment and prevention of a variety of thrombotic conditions, particularly coronary artery and cerebrovascular disease. Those experienced in this field are readily aware of the circumstances requiring anticoagulant therapy.
The term "patient" used herein is taken to mean mammals such as primates, including humans, sheep, horses, cattle, pigs, dogs, cats, rats, and mice.
Thrombin inhibition is useful not only in the anticoagulant therapy of individuals having thrombotic conditions, but is useful whenever inhibition of blood coagulation is required such as to prevent coagulation of stored whole blood and to prevent coagulation in other biological samples for testing or storage. Thus, thrombin inhibitors can be added to or contacted with any medium containing or suspected of containing thrombin and in which it is desired that blood coagulation be inhibited, e.g., when contacting the mammal's blood with material selected from the group consisting of vascular grafts, stents, orthopedic prothesis, cardiac prosthesis, and extracorporeal circulation systems Compounds of the invention are useful for treating or preventing venous thromboembolism (e.g. obstruction or occlusion of a vein by a detached thrombus; obstruction or occlusion of a lung artery by a detached thrombus), cardiogenic thromboembolism (e.g. obstruction or occlusion of the heart by a detached thrombus), arterial thrombosis (e.g.
formation of a thrombus within an artery that may cause infarction of tissue supplied by the artery), atherosclerosis (e.g. arteriosclerosis characterized by irregularly distributed lipid deposits) in mammals, and for lowering the propensity of devices that come into contact with blood to clot blood.
Examples of venous thromboembolism which may be treated or prevented with compounds of the invention include obstruction of a vein, obstruction of a lung artery (pulmonary embolism), deep vein thrombosis, thrombosis associated with cancer and cancer chemotherapy, thrombosis inherited with thrombophilic diseases such as Protein C deficiency, Protein S deficiency, antithrombin III
deficiency, and Factor V Leiden, and thrombosis resulting from acquired thrombophilic disorders such as systemic lupus erythematosus (inflammatory connective tissue disease). Also with regard to venous thromboembolism, compounds of the invention are useful for maintaining patency of indwelling catheters.
Examples of cardiogenic thromboembolism which may be treated or prevented with compounds of the invention include thromboembolic stroke (detached thrombus causing neurological affliction related to impaired cerebral blood supply), cardiogenic thromboembolism associated with atrial fibrillation (rapid, irregular twitching of upper heart chamber muscular fibrils), cardiogenic thromboembolism associated with prosthetic heart valves such as mechanical heart valves, and cardiogenic thromboembolism associated with heart disease.
Examples of arterial thrombosis include unstable angina (severe constrictive pain in chest of coronary origin), myocardial infarction (heart muscle cell death resulting from insufficient blood supply), ischemic heart disease (local anemia due to obstruction (such as by arterial narrowing} of blood supply), reocclusion during or after percutaneous transluminal coronary angioplasty, restenosis after percutaneous transluminal coronary angioplasty, occlusion of coronary artery bypass grafts, and occlusive cerebrovascular disease. Also with regard to arterial thrombosis, compounds of the invention are useful for maintaining patency in arteriovenous cannulas.
Examples of atherosclerosis include arteriosclerosis.
Examples of devices that come into contact with blood include vascular grafts, stents, orthopedic prosthesis, cardiac prosthesis, and extracorporeal circulation systems The thrombin inhibitors of the invention can be administered in such oral forms as tablets, capsules (each of which includes sustained release or timed release formulations), pills, powders, granules, elixers, tinctures, suspensions, syrups, and emulsions. Likewise, they may be administered in intravenous (bolus or infusion), intraperitoneal, subcutaneous, or intramuscular form, all using forms well known to those of ordinary skill in the pharmaceutical arts. An effective but non-toxic amount of the compound desired can be employed as an anti-aggregation agent. For treating ocular build up of fibrin, the compounds may be administered intraocularly or topically as well as orally or parenterally.
The thrombin inhibitors can be administered in the form of a depot injection or implant preparation which may be formulated in such a manner as to permit a sustained release of the active ingredient.
The active ingredient can be compressed into pellets or small cylinders and implanted subcutaneously or intramuscularly as depot injections or implants. Implants may employ inert materials such as biodegradable polymers or synthetic silicones, for example, Silastic, silicone rubber or other polymers manufactured by the Dow-Corning Corporation.
The thrombin inhibitors can also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles. Liposomes can be formed from a variety of phospholipids, such as cholesterol, stearylamine or phosphatidylcholines.
The thrombin inhibitors may also be delivered by the use of monoclonal antibodies as individual carriers to which the compound molecules are coupled. The thrombin inhibitors may also be coupled with soluble polymers as targetable drug carriers. Such polymers can include polyvinlypyrrolidone, pyran copolymer, polyhydroxy-propyl-methacrylamide-phenol, polyhydroxyethyl-aspartamide-phenol, or polyethyleneoxide-polylysine substituted with palinitoyl residues.
Furthermore, the thrombin inhibitors may be coupled to a class of biodegradable polymers useful in achieving controlled release of a drug, 5 for example, polylactic acid, polyglycolic acid, copolymers of polylactic and polyglycolic acid, polyepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates and cross linked or amphipathic block copolymers of hydrogels.
The dosage regimen utilizing the thrombin inhibitors is selected in accordance with a variety of factors including type, species, age, weight, sex and medical condition of the patient; the severity of the condition to be treated; the route of administration; the renal and hepatic function of the patient; and the particular compound or salt thereof employed. An ordinarily skilled physician or veterinarian can readily 15 determine and prescribe the ei~ective amount of the drug required to prevent, counter, or arrest the progress of the condition.
Oral dosages of the thrombin inhibitors, when used for the indicated effects, will range between about 0.01 mg per kg of body weight per day (mg/kg/day) to about 30 mg/kg/day, preferably 0.025-7.5 20 mg/kg/day, more preferably 0.1-2.5 mg/kg/day, and most preferably 0.1-0.5 mg/kg/day (unless specified otherwise, amounts of active ingredients are on free base basis). For example, an 80 kg patient would receive between about 0.8 mg/day and 2.4 g/day, preferably 2-600 mg/day, more preferably 8-200 mg/day, and most preferably 8-40 mg/kg/day. A suitably 25 prepared medicament for once a day administration would thus contain between 0.8 mg and 2.4 g, preferably between 2 mg and 600 mg, more preferably between 8 mg and 200 mg, and most preferably 8 mg and 40 mg, e.g., 8 mg, 10 mg, 20 mg and 40 mg. Advantageously, the thrombin inhibitors may be administered in divided doses of two, three, or four 30 times daily. For administration twice a day, a suitably prepared medicament would contain between 0.4 mg and 4 g, preferably between 1 mg and 300 mg, more preferably between 4 mg and 100 mg, and most preferably 4 mg and 20 mg, e.g., 4 mg, 5 mg, 10 mg and 20 mg.
Intravenously, the patient would receive the active 35 ingredient in quantities suff cient to deliver between 0.025-7.5 mg/kg/day, preferably 0.1-2.5 mg/kg/day, and more preferably 0.1-0.5 mg/kg/day.
Such quantities may be administered in a number of suitable ways, e.g.
large volumes of low concentrations of active ingredient during one extended period of time or several times a day, low volumes of high concentrations of active ingredient during a short period of time, e.g.
once a day. Typically, a conventional intravenous formulation may be prepared which contains a concentration of active ingredient of between about O.OI-1.0 mg/ml, e.g. 0.1 mg/ml, 0.3 mg/ml, and 0.6 mg/ml, and administered in amounts per day of between 0.01 ml/kg patient weight and 10.0 ml/kg patient weight, e.g. 0.1 ml/kg, 0.2 ml/kg, 0.5 ml/kg. In one example, an 80 kg patient, receiving 8 ml twice a day of an intravenous formulation having a concentration of active ingredient of 0.5 mg/ml, receives 8 mg of active ingredient per day. Glucuronic acid, L-lactic acid, acetic acid, citric acid or any pharmaceutically acceptable acid/conjugate base with reasonable buffering capacity in the pH range acceptable for intravenous administration may be used as buffers.
Consideration should be given to the solubility of the drug in choosing an The choice of appropriate buffer and pH of a formulation, depending on solubility of the drug to be administered, is readily made by a person having ordinary skill in the art.
The compounds can also be administered in intranasal form via topical use of suitable intranasal vehicles, or via transdermal routes, using those forms of transdermal skin patches well known to those of ordinary skill in that art. To be administered in the form of a transdermal delivery system, the dosage administration will, or course, be continuous rather than intermittent throughout the dosage regime.
The thrombin inhibitors are typically administered as active ingredients in admixture with suitable pharmaceutical diluents, excipients or carriers (collectively referred to herein as "carrier"
materials) suitably selected with respect to the intended form of administration, that is, oral tablets, capsules, elixers, syrups and the like, and consistent with convention pharmaceutical practices.
For instance, for oral administration in the form of a tablet or capsule, the active drug component can be combined with an oral, non-toxic, pharmaceutically acceptable, inert carrier such as lactose, starch, sucrose, glucose, methyl cellulose, magnesium stearate, dicalcium phosphate, calcium sulfate, mannitol, sorbitol and the like;
for oral administration in liquid form, the oral drug components can be combined with any oral, non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water and the like. Moreover, when desired or necessary, suitable binders, lubricants, disintegrating agents and coloring agents can also be incorporated into the mixture. Suitable binders include starch, gelatin, natural sugars such as glucose or beta-lactose, corn-sweeteners, natural and synthetic gums such as acacia, tragacanth or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes and the like. Lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like. Disintegrators include, without limitation, starch methyl cellulose, agar, bentonite, xanthan gum and the like.
Typical tablet cores suitable for administration of thrombin inhibitors are comprised of, but not limited to, the following amounts of standard ingredients:
Suggested Ranges of Composition for Excipients in Uncoated Tablet Cores Excipient General Preferred Most Preferred Range (%) Range (%) Range (%) mannitol 10-90 25-75 3p.sp microcrystalline 10-90 25-?5 30-60 cellulose magnesium 0.1-5.0 0.1-2.5 0.5-1.5 stearate Mannitol, microcrystalline cellulose and magnesium stearate may be substituted with alternative pharmaceutically acceptable excipients.
The thrombin inhibitors can also be co-administered with suitable anti-platelet agents, including, but not limited to, aspirin and fibrinogen receptor antagonists (e.g. to treat or prevent unstable angina or to prevent reocclusion after angioplasty and restenosis), anticoagulants such as heparin or warfarin, thrombolytic agents such as plasminogen activators or streptokinase to achieve synergistic effects in the treatment of various vascular pathologies, or lipid lowering agents including antihypercholesterolemics (e.g. HMG CoA reductase inhibitors such as lovastatin, simvastatin, HMG CoA synthase inhibitors, etc.) to treat or prevent atherosclerosis, or other agents that exert an antiatherogenic effect such as but not limited to insulin sensitizers and profibrinolytic agents including but not limited to agents that 1) increase levels and/or activity of one or more of the following proteins: tissue plasminogen activator, urokinase, plasmin, and/or 2) decrease levels and/or activity of one or more of the following proteins:
plasminogen activator inhibitor-1, thrombin activated fibrinolysis inhibitor and fibrinogen.
For example, patients suffering from coronary artery disease, and patients subjected to angioplasty procedures, would benefit from coadministration of fibrinogen receptor antagonists and thrombin inhibitors. Also, thrombin inhibitors enhance the efficiency of tissue plasminogen activator-mediated thrombolytic reperfusion. Thrombin inhibitors may be administered first following thrombus formation, and tissue plasminogen activator or other plasminogen activator is administered thereafter.
Typical doses of thrombin inhibitors of the invention in combination with other suitable anti-platelet agents, anticoagulation agents, or thrombolytic agents may be the same as those doses of thrombin inhibitors administered without coadminiatration of additional anti-platelet agents, anticoagulation agents, or thrombolytic agents, or may be substantially less that those doses of thrombin inhibitors administered without coadministration of additional anti-platelet agents, anticoagulation agents, or thrombolytic agents, depending on a patient's therapeutic needs.
The following synthetic methods can be used to prepare the compounds of the present invention:
The following examples are illustrative of the invention as contemplated by the inventors and should not be construed as being limits on the scope or spirit of the instant invention.
Amide couplings used to form the compounds of this invention are typically performed by the carbodiimide method with reagents such as dicyclohexylcarbodiimide, or 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 synthesis by classical Merrifield techniques may be employed instead.
The addition and removal of one or more protecting groups is also typical practice.
Compounds of the invention can be prepared according to the general procedure outlined below:
WO 99!16750 PCT/US98/20174 R O
1. DPPC/Et3N R O
BocNH~~~~OH ~, ~~
2. 4-aminopyridine BocNH~~~NH
N
R O
N H '~~~ N H
y N
LiAIH4 R O R
H
NH'~~NH EDC/HOBT R2~N N NH
H
o R
J
~N~ R~ N OH
N
Preparation of intermediate N
f Step A:
O
BocN H O~ I-1 A -10°C solution of N-t-Boc-L-Phe (10.6 g, 40.0 mmol) in 350 mL of EtOAc was treated with 4.4 mL (40 mmol) of N-methylmorpholine then 5.2 mL (40 mmol) of isobutyl chloroformate. The cold solution was 10 stirred for lh before it was quenched by the addition of 100 mL of water.
The organic phase was collected and dried (MgS04). To the resulting solution containing the mixed anhydride was added 180 mL of ethereal CH2N2 by pipet and the whole was stirred at room temperature far 2 h.
Excess diazomethane was removed by purging the solution with N2 15 before removing the solvent in uaccuo. The obtained oil was redissalved in 500 mL of absolute EtOH containing 6.8 mL (48 mmol) of Et3N. The mixture was carefully treated with 2.8 g (12 mmol) of silver benzoate which resulted in the vigorous release of N2. The black solution was stirred at room temperature for 2 h, concentrated and chromatographed 20 (9:1 Hexane / EtOAc) to afford the desired ester ,I-~ as a yellow oil.
1H NMR (CDC13) S 7.2-7.4 (m, 5H), 5.05 (m, 1H), 4.19 (q, J=7.2 Hz, 2H), 2.85 (m, 2H), 2.43 (m, 2H), 1.40 (s, 9H), 1.25 (t, J=7.2 Hz, 3H).
\
BocN H O H I-2 A solution of I-,~, (1.5 g, 4.9 mmol) in 50 mL of THF and 50 mL of water was treated with 1.05 g (25 mmol ) of LiOH ~ H20 and the mixture was stirred for 15h before removing the solvent in vacuo. The obtained salt was redissolved in 100 mL of absolute EtOAc the mixture was acidified with 30 mL of lj~ HCl. The organic phase was separated, washed with brine, dried (MgS04) and concentrated to afford the acid _I-1H NMR (CDC13) 8 ?.2-7.4 (m, 5H), 5.05 (m, 1H), 4.19 (m, IH), 2.80-3.00 (m, 2H), 2.40-2.55 (m, 2H), 1.40 (s, 9H).
Sten CC:
\ O / N
~ ~ \
BocNH- v 'N
A solution of ~, (13.5 g, 48.3 mmol) in 300 mL of THF at 0°C
was treated with 8.0 mL (58.0 mmol) of Et3N then 7.00 mL (53.1 mmol) of 20 diphenylphoshinic chloride. The mixture was stirred for 45 min before the addition of 9.0 g (9fi.7 mmol) of 4-aminopyridine. The reaction mixture was stirred to room temperature over 3h, diluted with 300 mL of EtOAc and quenched with 50 mL of 5% Na2C03. The organic phase was -separated, washed with saturated NH4C1 (3 x 50 mL), dried (MgS04) and concentrated to afford the desired amide ~.
1H NMR (CDCIg) b 8.65 (bs,1H), 8.40 (d, J=6.8 Hz, 2H), 7.48 (d, J=6.8 Hz, 5 2H), 7.2-7.4 (m, 5H), 5.15 (bd, 1H), 4.19 (m, 1H), 2.80-3.00 (m, 2H), 2.5-2.57 (m, 2H), 1.40 (s, 9H).
O ~ ~N
t H I_4 to To a 0°C solution of ~ (14 g, 39.4 mmol) in 90 mL of CH2C12 was added 90 mL of TFA. The mixture was stirred for 2h and concentrated to dryness. The obtained oil was redissolved in 200 mL of EtOAc and thrice extracted with 50 mL of 5% Na2C03. The organic 15 phase was dried (MgS04) and concentrated to afford the above amine ~.
1H NMR (CDCl3) 8 8.40 (d, J=6.8 Hz, 2H), 7.40 (d, J=6.8 Hz, 2H), 7.1-7.3 (m, 5H) 3.4 (m, 1H), 2.85 (m, 2H), 2.60 (m, 2H), 2.35 (m, 1H).
N
A solution of ~ (4.25 g, 16.7 mmol) in 300 mL of THF at 0°C
was carefully treated with 100 mL of LiAlH4 by dropping funnel. The mixture was stirred to room temperature over 20h and quenched by the dropwise addition (at 0°C) of 10 mL of water, 10 mL of 15% NaOH, and 30 mL of water. The reaction mixture was diluted with 200 mL of EtOAc and the solids were removed by filtration. The filtrate was washed with 10 saturated NaK tartrate (2 x 50 mL) and brine. The organic phase was dried (MgS04) and concentrated to afford a yellow oil which was chromatographed (5% MeOH in CHC13 saturated with NH3) to afford the above diamine ~.
15 1H NMR (CDC13) 8 8.15 (d, J=6.8 Hz, 2H), 7.I-7.3 (m, 5H), 6.40 (d, J=6.8 Hz, 2H), 5.30 ~(bs, 1H), 3.2-3.4 (m, 2H), 3.15 (m, 1H), 2.80 (m, 1H), 2.55 (m, 1H), 1.85 (m, 1H), 1.58 (m, 1H).
_~5_ /
H O ~ / N
' ~ I
,S; N v _N N
O O H H
Steu l1:
H O
,S; N v _OH
/ O O
Glycine methyl ester was prepared by the addition of 2.8 mL
(20 mmol) of Et3N to 2.5 g (20 mmol) of the amino ester HCl salt in 50 mL
of DCM. After stirring for 10 min, 5.0 mL (60 mmol) of pyridine was added followed by 2.85 g (15 mmol) of a-toluenesulfonyl chloride. The reaction mixture was stirred for 1 h, diluted with 100 mL of DCM then quenched with water. The organic phase was separated and washed 15 with 5% Na2C03 (3 x 10 mL), lj~ HCl (3 x 10 mL) and brine.
Concentration of the organic phase afforded the crude ester (4.5 g, 18.5 mmol) which was dissolved in 21 mL of dioxane and 7 mL of water and treated with 3.0 g (71.4 mmol) of LiOH~H20. This new reaction mixture was stirred at room temperature for 16 h, concentrated and acidified to 20 pH=1 with lj~ HCl. The solid was dissolved in EtOAc and the organic phase was separated and washed with brine. The solution was dried and concentrated to afford N-benzylsulfonylglycine 1,~1- as a white solid.
1H NMR (DMSO-d6) 8 7.45 (m, 2H), 7.38 (m, 3H), 6.80 (bs, 1H), 4.35 (s, 2H), 3.63 (d, J=6 Hz, 2H).
Steu 22:
H ' / N
I
,N
,S, N N
O O H H
To a solution of the diamine ,~ (140 mg, 0.62 mmol) in 2 mL
of DMF was added 140 mg (0.62 mmol) of N-a-toluenesulfonylglycine (~
~ 83 mg (0.62 mmol) of HOBT, 118 mg (0.62 mmol) of EDC and 0.22 mL
(1.2 mmol) of DIPEA. The mixture was stirred to room temperature over 16 h, concentrated and chromatographed ( 10% MeOH in CHC13 saturated with NH3) to afford ,~ as a white solid.
1H NMR (CDC13) b 8.I5 (d, J=6.8 Hz, 2H), 7.1-7.5 (m, 10H), 6.95 (d, 16 Hz, 1H), 6.40 (d, J=6.8 Hz, 2H), 5.20 (bs, 1H), 4.25 (s, 2H), 3.51 (s, 2H), 3.0-3.3 (m, 2H), 2.84 (t, 2H), 1.85 (m, 1H), 1.58 (m, 1H).
Anal. Calc'd for C24H28N4S03: C; 63.69, H; 6.24, N; 12.38. Found: C;
63.42, H; 6.24, N; 12.17.
/ I
CI /
I H O \ / N
' ~ \ I
\ ,S~ N v _N N
O ~O H H
2 ~ was prepared in an analogous fashion as described in example 1 starting from 4-chlorobenzensulfonyl chloride and glycine methyl ester.
/ I
CI /
I H O \ / N
' ~ \ I
CI \ ,S~ N v 'N N
O O H H
~ was prepared in an analogous fashion as described in example 1 starting from 3,4-dichlorobenzensulfonyl chloride and glycine methyl ester.
/ I
/ / I H O ~ / N
S; N~ N N
O ~O
~1, was prepared in an analogous fashion as described in example 1 starting from 2-naphthalenesulfonyl chloride and glycine methyl ester.
/ I
CH30 / H O ~ /
_N
~ i N~ ~ I
;S~ N N
O O
,~ was prepared in an analogous fashion as described in example 1 starting from 4-methoxy-2,3,6-trimethylbenzenesulfonyl chloride and glycine methyl ester.
/
H O ~ / N
I' ~
,S; N v ' N N
~ i / O O ~ H H
OH 6_1 ~, was prepared in an analogous fashion as described in example 1 starting from a-toluenesulfonyl chloride and L-serine methyl ester.
H O ~ / N
I
~N~
;S, _ N N
Specific exemplifications of this class are shown in the table below Table 1 H O R
R2~N N N~H
R~ H n N
R n R2 R1 Ki(~,M) Bn 0 Boc H
Bn 0 PhS02- H
c-hexylCH2- 0 PhS02- H
Bn 0 4-ClPhS02- H
Bn 1 4-Cl-PhS02- H **
Bn 1 B~2- H **
Bn 1 2-naphthS02- H **
Bn 1 3,4-di-Cl-PhS02-H **
Bn 1 PhCH=CHS02- H **
Bn 1 4-Cl-BnS02- H **
Bn 1 4-CF3-BnS02- H **
Bn 1 3,4-di-Cl-BnS02-H **
Bn 1 4-OCH3-2,3,6- H **
MePhS02-Bn 1 3,4-di-Cl-PhS02-(S)-CH20H **
* > 1.0 E.~M
**<_1.ONM
_7_ Tabl 1 ( ,nnt'rl) H O R
R2~N N N~H
R1 H n y N
R n R2 R1 ~(~) Bn 1 BnS02- (R)-CH3 Bn 1 BnS02- (S)-CHg **
Bn 1 BnS02- (S)-CH2CH2S02CH3 **
Bn 1 BnS02- (S)-CH2CH2SCHg **
Bn 1 H 4-CIBn- **
(CH3)2CH2- 1 BnS02- H
(CHg)2CH2- I 3,4-di-Cl-PhS02-H **
* > 1.0 N,M
** 5 L0 N,M
Exemplifications also include the pharmaceutically acceptable salts of the above-identified compounds.
_g_ ABBREVIATIONS
Desienation Protecting =ro m BOC (Boc) t-butyloxycarbonyl CBZ (Cbz) benzyloxycarbonyl(carbobenzoxy) Ph phenyl Bn benzyl TBS (TBDMS) t-butyl-dimethylsilyl Activating GrOUD
HBT(HOBT or HOBt) 1-hydroxybenzotriazole hydrate Designation ~Dli_n_~ Reagg~t BOP reagent benzotriazol-1-yloxytris-(dimethylamino)phosphonium hexafluorophosphate BOP-Cl bis(2-oxo-3-oxazolidinyl)phosphinic chloride EDC 1-ethyl-3-(3-dimethylaminopropyl ) carbodiimide hydrochloride _g_ Other (BOC~O (BOC20) di-t-butyl dicarbonate n-Bu4N+F- tetrabutyl ammonium fluoride nBuLi (n-Buli) n-butyllithium DMF dimethylformamide EtgN (TEA) triethylamine EtOAc ethyl acetate TFA trifluoroacetic acid DMAP dimethylaminopyridine DME dimethoxyethane NMM N-methylmorpholine DPPA diphenylphosphoryl azide THF tetrahydrofuran DPPC diphenylphosphinic chloride DIPEA diisopropylethylamine Amino Acid Ile Isoleucine Phe Phenylalanine Pro Proline Ala Alanine V al Valine 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. The compounds of the present invention may also have polymorphic crystalline forms, with all polymorphic crystalline forms being included in the present invention.
10 When any variable occurs more than one time in any constituent or in formula I, its definition on each occurrence is independent of its definition at every other occurrence. Also, combinations of substituents and/or variables are permissible only if such combinations result in stable compounds.
As used herein, except where noted, "alkyl" is intended to include both branched- and straight-chain saturated aliphatic hydrocarbon groups having, unless otherwise noted, 1-8 carbon atoms (Me is methyl, Et is ethyl, Pr is propyl, Bu is butyl). "Alkenyl" is 5 intended to include both branched- and straight-chain unsaturated aliphatic hydrocarbon groups having, unless otherwise noted, 1-8 carbon atoms, e.g. ethenyl, propenyl, etc. "Cycloalkyl" includes cyclic saturated aliphatic hydrocarbon groups having 3-8 carbon atoms (e.g. "C3_ gcycloalkyl" is intended to include cyclopropyl, cyclobutyl, cyclopentyl, 10 cyclohexyl and cycloheptyl, and the like). The term "C7-12 bicyclic alkyl"
is intended to include bicyclo[2.2.1]heptyl (norbornyl), bicyclo[2.2.2]octyl, 1,1,3-trimethyl-bicyclo[2.2.1]heptyl (bornyl), and the like. The term "aryl" as used herein except where noted, represents a stable 6- to 10-membered mono- or bicyclic ring system such as phenyl, or naphthyl:
15 The aryl ring can be unsubstituted or substituted with one or more of C1_ 4 lower alkyl; hydroxy; alkoxy; halogen; amino. The term "heteroaryl"
refers to a 5- to 7- membered unsaturated ring containing 1 or 2 heteroatoms selected from O, N, or S. "Alkoxy" represents an alkyl group having 1-8 carbon atoms attached through an oxygen bridge.
20 "Halo", as used herein, means fluoro, chloro, bromo and iodo.
"Counteriori' is used to represent a small, single negatively-charged species, such as chloride, bromide, hydroxide, acetate, trifluroacetate, perchlorate, nitrate, benzoate, maleate, tartrate, hemitartrate, benzene sulfonate, and the like.
25 Under standard nomenclature used throughout this disclosure unless specified otherwise, the terminal portion of the designated side chain is described first followed by the adjacent functionality toward the point of attachment. For example, an ethyl substituent substituted with "methylcarbonylamino" is equivalent to O
The term "heterocycle" or "heterocyclic ring", as used herein except where noted, represents a stable 5- to 7-membered mono-or bicyclic or stable 7- to 10-membered bicyclic 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 of N, O and S, and wherein the nitrogen and sulfur heteroatoms may optionally be oxidized, and the nitrogen heteroatom may optionally be quaternized, and including any bicyclic group in which any of the above-defined heterocyclic rings is fused to a benzene ring. Especially useful are rings containing one oxygen or sulfur, one to four nitrogen atoms, or one oxygen or sulfur combined with one or two nitrogen atoms. The heterocyclic ring may be attached at any heteroatom or carbon atom which results in the creation of a stable structure. Examples of such heterocyclic elements include piperidinyl, piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolodinyl, 2-oxoazepinyl, azepinyl, pyrrolyl, 4-piperidonyl, 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, thiamorpholinyl sulfone, and oxadiazolyl. Morpholino is the same as 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 salts or the quaternary ammonium salts which are formed, e.g., from inorganic or organic acids or bases.
Examples of such acid addition salts include acetate, adipate, alginate, 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, 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 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, and butyl chloride, bromides and iodides;
dialkyl sulfates like dimethyl, diethyl, dibutyl and diamyl sulfates, long chain halides such as decyl, Iauryl, myristyl and stearyl chlorides, bromides and iodides, aralkyl halides Iike benzyl and phenethyl bromides and others.
Thrombin Inhibitors - Thera;~pmt;r~. Tl~eP~
Anticoagulant therapy is indicated for the treatment and prevention of a variety of thrombotic conditions, particularly coronary artery and cerebrovascular disease. Those experienced in this field are readily aware of the circumstances requiring anticoagulant therapy.
The term "patient" used herein is taken to mean mammals such as primates, including humans, sheep, horses, cattle, pigs, dogs, cats, rats, and mice.
Thrombin inhibition is useful not only in the anticoagulant therapy of individuals having thrombotic conditions, but is useful whenever inhibition of blood coagulation is required such as to prevent coagulation of stored whole blood and to prevent coagulation in other biological samples for testing or storage. Thus, thrombin inhibitors can be added to or contacted with any medium containing or suspected of containing thrombin and in which it is desired that blood coagulation be inhibited, e.g., when contacting the mammal's blood with material selected from the group consisting of vascular grafts, stents, orthopedic prothesis, cardiac prosthesis, and extracorporeal circulation systems Compounds of the invention are useful for treating or preventing venous thromboembolism (e.g. obstruction or occlusion of a vein by a detached thrombus; obstruction or occlusion of a lung artery by a detached thrombus), cardiogenic thromboembolism (e.g. obstruction or occlusion of the heart by a detached thrombus), arterial thrombosis (e.g.
formation of a thrombus within an artery that may cause infarction of tissue supplied by the artery), atherosclerosis (e.g. arteriosclerosis characterized by irregularly distributed lipid deposits) in mammals, and for lowering the propensity of devices that come into contact with blood to clot blood.
Examples of venous thromboembolism which may be treated or prevented with compounds of the invention include obstruction of a vein, obstruction of a lung artery (pulmonary embolism), deep vein thrombosis, thrombosis associated with cancer and cancer chemotherapy, thrombosis inherited with thrombophilic diseases such as Protein C deficiency, Protein S deficiency, antithrombin III
deficiency, and Factor V Leiden, and thrombosis resulting from acquired thrombophilic disorders such as systemic lupus erythematosus (inflammatory connective tissue disease). Also with regard to venous thromboembolism, compounds of the invention are useful for maintaining patency of indwelling catheters.
Examples of cardiogenic thromboembolism which may be treated or prevented with compounds of the invention include thromboembolic stroke (detached thrombus causing neurological affliction related to impaired cerebral blood supply), cardiogenic thromboembolism associated with atrial fibrillation (rapid, irregular twitching of upper heart chamber muscular fibrils), cardiogenic thromboembolism associated with prosthetic heart valves such as mechanical heart valves, and cardiogenic thromboembolism associated with heart disease.
Examples of arterial thrombosis include unstable angina (severe constrictive pain in chest of coronary origin), myocardial infarction (heart muscle cell death resulting from insufficient blood supply), ischemic heart disease (local anemia due to obstruction (such as by arterial narrowing} of blood supply), reocclusion during or after percutaneous transluminal coronary angioplasty, restenosis after percutaneous transluminal coronary angioplasty, occlusion of coronary artery bypass grafts, and occlusive cerebrovascular disease. Also with regard to arterial thrombosis, compounds of the invention are useful for maintaining patency in arteriovenous cannulas.
Examples of atherosclerosis include arteriosclerosis.
Examples of devices that come into contact with blood include vascular grafts, stents, orthopedic prosthesis, cardiac prosthesis, and extracorporeal circulation systems The thrombin inhibitors of the invention can be administered in such oral forms as tablets, capsules (each of which includes sustained release or timed release formulations), pills, powders, granules, elixers, tinctures, suspensions, syrups, and emulsions. Likewise, they may be administered in intravenous (bolus or infusion), intraperitoneal, subcutaneous, or intramuscular form, all using forms well known to those of ordinary skill in the pharmaceutical arts. An effective but non-toxic amount of the compound desired can be employed as an anti-aggregation agent. For treating ocular build up of fibrin, the compounds may be administered intraocularly or topically as well as orally or parenterally.
The thrombin inhibitors can be administered in the form of a depot injection or implant preparation which may be formulated in such a manner as to permit a sustained release of the active ingredient.
The active ingredient can be compressed into pellets or small cylinders and implanted subcutaneously or intramuscularly as depot injections or implants. Implants may employ inert materials such as biodegradable polymers or synthetic silicones, for example, Silastic, silicone rubber or other polymers manufactured by the Dow-Corning Corporation.
The thrombin inhibitors can also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles. Liposomes can be formed from a variety of phospholipids, such as cholesterol, stearylamine or phosphatidylcholines.
The thrombin inhibitors may also be delivered by the use of monoclonal antibodies as individual carriers to which the compound molecules are coupled. The thrombin inhibitors may also be coupled with soluble polymers as targetable drug carriers. Such polymers can include polyvinlypyrrolidone, pyran copolymer, polyhydroxy-propyl-methacrylamide-phenol, polyhydroxyethyl-aspartamide-phenol, or polyethyleneoxide-polylysine substituted with palinitoyl residues.
Furthermore, the thrombin inhibitors may be coupled to a class of biodegradable polymers useful in achieving controlled release of a drug, 5 for example, polylactic acid, polyglycolic acid, copolymers of polylactic and polyglycolic acid, polyepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates and cross linked or amphipathic block copolymers of hydrogels.
The dosage regimen utilizing the thrombin inhibitors is selected in accordance with a variety of factors including type, species, age, weight, sex and medical condition of the patient; the severity of the condition to be treated; the route of administration; the renal and hepatic function of the patient; and the particular compound or salt thereof employed. An ordinarily skilled physician or veterinarian can readily 15 determine and prescribe the ei~ective amount of the drug required to prevent, counter, or arrest the progress of the condition.
Oral dosages of the thrombin inhibitors, when used for the indicated effects, will range between about 0.01 mg per kg of body weight per day (mg/kg/day) to about 30 mg/kg/day, preferably 0.025-7.5 20 mg/kg/day, more preferably 0.1-2.5 mg/kg/day, and most preferably 0.1-0.5 mg/kg/day (unless specified otherwise, amounts of active ingredients are on free base basis). For example, an 80 kg patient would receive between about 0.8 mg/day and 2.4 g/day, preferably 2-600 mg/day, more preferably 8-200 mg/day, and most preferably 8-40 mg/kg/day. A suitably 25 prepared medicament for once a day administration would thus contain between 0.8 mg and 2.4 g, preferably between 2 mg and 600 mg, more preferably between 8 mg and 200 mg, and most preferably 8 mg and 40 mg, e.g., 8 mg, 10 mg, 20 mg and 40 mg. Advantageously, the thrombin inhibitors may be administered in divided doses of two, three, or four 30 times daily. For administration twice a day, a suitably prepared medicament would contain between 0.4 mg and 4 g, preferably between 1 mg and 300 mg, more preferably between 4 mg and 100 mg, and most preferably 4 mg and 20 mg, e.g., 4 mg, 5 mg, 10 mg and 20 mg.
Intravenously, the patient would receive the active 35 ingredient in quantities suff cient to deliver between 0.025-7.5 mg/kg/day, preferably 0.1-2.5 mg/kg/day, and more preferably 0.1-0.5 mg/kg/day.
Such quantities may be administered in a number of suitable ways, e.g.
large volumes of low concentrations of active ingredient during one extended period of time or several times a day, low volumes of high concentrations of active ingredient during a short period of time, e.g.
once a day. Typically, a conventional intravenous formulation may be prepared which contains a concentration of active ingredient of between about O.OI-1.0 mg/ml, e.g. 0.1 mg/ml, 0.3 mg/ml, and 0.6 mg/ml, and administered in amounts per day of between 0.01 ml/kg patient weight and 10.0 ml/kg patient weight, e.g. 0.1 ml/kg, 0.2 ml/kg, 0.5 ml/kg. In one example, an 80 kg patient, receiving 8 ml twice a day of an intravenous formulation having a concentration of active ingredient of 0.5 mg/ml, receives 8 mg of active ingredient per day. Glucuronic acid, L-lactic acid, acetic acid, citric acid or any pharmaceutically acceptable acid/conjugate base with reasonable buffering capacity in the pH range acceptable for intravenous administration may be used as buffers.
Consideration should be given to the solubility of the drug in choosing an The choice of appropriate buffer and pH of a formulation, depending on solubility of the drug to be administered, is readily made by a person having ordinary skill in the art.
The compounds can also be administered in intranasal form via topical use of suitable intranasal vehicles, or via transdermal routes, using those forms of transdermal skin patches well known to those of ordinary skill in that art. To be administered in the form of a transdermal delivery system, the dosage administration will, or course, be continuous rather than intermittent throughout the dosage regime.
The thrombin inhibitors are typically administered as active ingredients in admixture with suitable pharmaceutical diluents, excipients or carriers (collectively referred to herein as "carrier"
materials) suitably selected with respect to the intended form of administration, that is, oral tablets, capsules, elixers, syrups and the like, and consistent with convention pharmaceutical practices.
For instance, for oral administration in the form of a tablet or capsule, the active drug component can be combined with an oral, non-toxic, pharmaceutically acceptable, inert carrier such as lactose, starch, sucrose, glucose, methyl cellulose, magnesium stearate, dicalcium phosphate, calcium sulfate, mannitol, sorbitol and the like;
for oral administration in liquid form, the oral drug components can be combined with any oral, non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water and the like. Moreover, when desired or necessary, suitable binders, lubricants, disintegrating agents and coloring agents can also be incorporated into the mixture. Suitable binders include starch, gelatin, natural sugars such as glucose or beta-lactose, corn-sweeteners, natural and synthetic gums such as acacia, tragacanth or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes and the like. Lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like. Disintegrators include, without limitation, starch methyl cellulose, agar, bentonite, xanthan gum and the like.
Typical tablet cores suitable for administration of thrombin inhibitors are comprised of, but not limited to, the following amounts of standard ingredients:
Suggested Ranges of Composition for Excipients in Uncoated Tablet Cores Excipient General Preferred Most Preferred Range (%) Range (%) Range (%) mannitol 10-90 25-75 3p.sp microcrystalline 10-90 25-?5 30-60 cellulose magnesium 0.1-5.0 0.1-2.5 0.5-1.5 stearate Mannitol, microcrystalline cellulose and magnesium stearate may be substituted with alternative pharmaceutically acceptable excipients.
The thrombin inhibitors can also be co-administered with suitable anti-platelet agents, including, but not limited to, aspirin and fibrinogen receptor antagonists (e.g. to treat or prevent unstable angina or to prevent reocclusion after angioplasty and restenosis), anticoagulants such as heparin or warfarin, thrombolytic agents such as plasminogen activators or streptokinase to achieve synergistic effects in the treatment of various vascular pathologies, or lipid lowering agents including antihypercholesterolemics (e.g. HMG CoA reductase inhibitors such as lovastatin, simvastatin, HMG CoA synthase inhibitors, etc.) to treat or prevent atherosclerosis, or other agents that exert an antiatherogenic effect such as but not limited to insulin sensitizers and profibrinolytic agents including but not limited to agents that 1) increase levels and/or activity of one or more of the following proteins: tissue plasminogen activator, urokinase, plasmin, and/or 2) decrease levels and/or activity of one or more of the following proteins:
plasminogen activator inhibitor-1, thrombin activated fibrinolysis inhibitor and fibrinogen.
For example, patients suffering from coronary artery disease, and patients subjected to angioplasty procedures, would benefit from coadministration of fibrinogen receptor antagonists and thrombin inhibitors. Also, thrombin inhibitors enhance the efficiency of tissue plasminogen activator-mediated thrombolytic reperfusion. Thrombin inhibitors may be administered first following thrombus formation, and tissue plasminogen activator or other plasminogen activator is administered thereafter.
Typical doses of thrombin inhibitors of the invention in combination with other suitable anti-platelet agents, anticoagulation agents, or thrombolytic agents may be the same as those doses of thrombin inhibitors administered without coadminiatration of additional anti-platelet agents, anticoagulation agents, or thrombolytic agents, or may be substantially less that those doses of thrombin inhibitors administered without coadministration of additional anti-platelet agents, anticoagulation agents, or thrombolytic agents, depending on a patient's therapeutic needs.
The following synthetic methods can be used to prepare the compounds of the present invention:
The following examples are illustrative of the invention as contemplated by the inventors and should not be construed as being limits on the scope or spirit of the instant invention.
Amide couplings used to form the compounds of this invention are typically performed by the carbodiimide method with reagents such as dicyclohexylcarbodiimide, or 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 synthesis by classical Merrifield techniques may be employed instead.
The addition and removal of one or more protecting groups is also typical practice.
Compounds of the invention can be prepared according to the general procedure outlined below:
WO 99!16750 PCT/US98/20174 R O
1. DPPC/Et3N R O
BocNH~~~~OH ~, ~~
2. 4-aminopyridine BocNH~~~NH
N
R O
N H '~~~ N H
y N
LiAIH4 R O R
H
NH'~~NH EDC/HOBT R2~N N NH
H
o R
J
~N~ R~ N OH
N
Preparation of intermediate N
f Step A:
O
BocN H O~ I-1 A -10°C solution of N-t-Boc-L-Phe (10.6 g, 40.0 mmol) in 350 mL of EtOAc was treated with 4.4 mL (40 mmol) of N-methylmorpholine then 5.2 mL (40 mmol) of isobutyl chloroformate. The cold solution was 10 stirred for lh before it was quenched by the addition of 100 mL of water.
The organic phase was collected and dried (MgS04). To the resulting solution containing the mixed anhydride was added 180 mL of ethereal CH2N2 by pipet and the whole was stirred at room temperature far 2 h.
Excess diazomethane was removed by purging the solution with N2 15 before removing the solvent in uaccuo. The obtained oil was redissalved in 500 mL of absolute EtOH containing 6.8 mL (48 mmol) of Et3N. The mixture was carefully treated with 2.8 g (12 mmol) of silver benzoate which resulted in the vigorous release of N2. The black solution was stirred at room temperature for 2 h, concentrated and chromatographed 20 (9:1 Hexane / EtOAc) to afford the desired ester ,I-~ as a yellow oil.
1H NMR (CDC13) S 7.2-7.4 (m, 5H), 5.05 (m, 1H), 4.19 (q, J=7.2 Hz, 2H), 2.85 (m, 2H), 2.43 (m, 2H), 1.40 (s, 9H), 1.25 (t, J=7.2 Hz, 3H).
\
BocN H O H I-2 A solution of I-,~, (1.5 g, 4.9 mmol) in 50 mL of THF and 50 mL of water was treated with 1.05 g (25 mmol ) of LiOH ~ H20 and the mixture was stirred for 15h before removing the solvent in vacuo. The obtained salt was redissolved in 100 mL of absolute EtOAc the mixture was acidified with 30 mL of lj~ HCl. The organic phase was separated, washed with brine, dried (MgS04) and concentrated to afford the acid _I-1H NMR (CDC13) 8 ?.2-7.4 (m, 5H), 5.05 (m, 1H), 4.19 (m, IH), 2.80-3.00 (m, 2H), 2.40-2.55 (m, 2H), 1.40 (s, 9H).
Sten CC:
\ O / N
~ ~ \
BocNH- v 'N
A solution of ~, (13.5 g, 48.3 mmol) in 300 mL of THF at 0°C
was treated with 8.0 mL (58.0 mmol) of Et3N then 7.00 mL (53.1 mmol) of 20 diphenylphoshinic chloride. The mixture was stirred for 45 min before the addition of 9.0 g (9fi.7 mmol) of 4-aminopyridine. The reaction mixture was stirred to room temperature over 3h, diluted with 300 mL of EtOAc and quenched with 50 mL of 5% Na2C03. The organic phase was -separated, washed with saturated NH4C1 (3 x 50 mL), dried (MgS04) and concentrated to afford the desired amide ~.
1H NMR (CDCIg) b 8.65 (bs,1H), 8.40 (d, J=6.8 Hz, 2H), 7.48 (d, J=6.8 Hz, 5 2H), 7.2-7.4 (m, 5H), 5.15 (bd, 1H), 4.19 (m, 1H), 2.80-3.00 (m, 2H), 2.5-2.57 (m, 2H), 1.40 (s, 9H).
O ~ ~N
t H I_4 to To a 0°C solution of ~ (14 g, 39.4 mmol) in 90 mL of CH2C12 was added 90 mL of TFA. The mixture was stirred for 2h and concentrated to dryness. The obtained oil was redissolved in 200 mL of EtOAc and thrice extracted with 50 mL of 5% Na2C03. The organic 15 phase was dried (MgS04) and concentrated to afford the above amine ~.
1H NMR (CDCl3) 8 8.40 (d, J=6.8 Hz, 2H), 7.40 (d, J=6.8 Hz, 2H), 7.1-7.3 (m, 5H) 3.4 (m, 1H), 2.85 (m, 2H), 2.60 (m, 2H), 2.35 (m, 1H).
N
A solution of ~ (4.25 g, 16.7 mmol) in 300 mL of THF at 0°C
was carefully treated with 100 mL of LiAlH4 by dropping funnel. The mixture was stirred to room temperature over 20h and quenched by the dropwise addition (at 0°C) of 10 mL of water, 10 mL of 15% NaOH, and 30 mL of water. The reaction mixture was diluted with 200 mL of EtOAc and the solids were removed by filtration. The filtrate was washed with 10 saturated NaK tartrate (2 x 50 mL) and brine. The organic phase was dried (MgS04) and concentrated to afford a yellow oil which was chromatographed (5% MeOH in CHC13 saturated with NH3) to afford the above diamine ~.
15 1H NMR (CDC13) 8 8.15 (d, J=6.8 Hz, 2H), 7.I-7.3 (m, 5H), 6.40 (d, J=6.8 Hz, 2H), 5.30 ~(bs, 1H), 3.2-3.4 (m, 2H), 3.15 (m, 1H), 2.80 (m, 1H), 2.55 (m, 1H), 1.85 (m, 1H), 1.58 (m, 1H).
_~5_ /
H O ~ / N
' ~ I
,S; N v _N N
O O H H
Steu l1:
H O
,S; N v _OH
/ O O
Glycine methyl ester was prepared by the addition of 2.8 mL
(20 mmol) of Et3N to 2.5 g (20 mmol) of the amino ester HCl salt in 50 mL
of DCM. After stirring for 10 min, 5.0 mL (60 mmol) of pyridine was added followed by 2.85 g (15 mmol) of a-toluenesulfonyl chloride. The reaction mixture was stirred for 1 h, diluted with 100 mL of DCM then quenched with water. The organic phase was separated and washed 15 with 5% Na2C03 (3 x 10 mL), lj~ HCl (3 x 10 mL) and brine.
Concentration of the organic phase afforded the crude ester (4.5 g, 18.5 mmol) which was dissolved in 21 mL of dioxane and 7 mL of water and treated with 3.0 g (71.4 mmol) of LiOH~H20. This new reaction mixture was stirred at room temperature for 16 h, concentrated and acidified to 20 pH=1 with lj~ HCl. The solid was dissolved in EtOAc and the organic phase was separated and washed with brine. The solution was dried and concentrated to afford N-benzylsulfonylglycine 1,~1- as a white solid.
1H NMR (DMSO-d6) 8 7.45 (m, 2H), 7.38 (m, 3H), 6.80 (bs, 1H), 4.35 (s, 2H), 3.63 (d, J=6 Hz, 2H).
Steu 22:
H ' / N
I
,N
,S, N N
O O H H
To a solution of the diamine ,~ (140 mg, 0.62 mmol) in 2 mL
of DMF was added 140 mg (0.62 mmol) of N-a-toluenesulfonylglycine (~
~ 83 mg (0.62 mmol) of HOBT, 118 mg (0.62 mmol) of EDC and 0.22 mL
(1.2 mmol) of DIPEA. The mixture was stirred to room temperature over 16 h, concentrated and chromatographed ( 10% MeOH in CHC13 saturated with NH3) to afford ,~ as a white solid.
1H NMR (CDC13) b 8.I5 (d, J=6.8 Hz, 2H), 7.1-7.5 (m, 10H), 6.95 (d, 16 Hz, 1H), 6.40 (d, J=6.8 Hz, 2H), 5.20 (bs, 1H), 4.25 (s, 2H), 3.51 (s, 2H), 3.0-3.3 (m, 2H), 2.84 (t, 2H), 1.85 (m, 1H), 1.58 (m, 1H).
Anal. Calc'd for C24H28N4S03: C; 63.69, H; 6.24, N; 12.38. Found: C;
63.42, H; 6.24, N; 12.17.
/ I
CI /
I H O \ / N
' ~ \ I
\ ,S~ N v _N N
O ~O H H
2 ~ was prepared in an analogous fashion as described in example 1 starting from 4-chlorobenzensulfonyl chloride and glycine methyl ester.
/ I
CI /
I H O \ / N
' ~ \ I
CI \ ,S~ N v 'N N
O O H H
~ was prepared in an analogous fashion as described in example 1 starting from 3,4-dichlorobenzensulfonyl chloride and glycine methyl ester.
/ I
/ / I H O ~ / N
S; N~ N N
O ~O
~1, was prepared in an analogous fashion as described in example 1 starting from 2-naphthalenesulfonyl chloride and glycine methyl ester.
/ I
CH30 / H O ~ /
_N
~ i N~ ~ I
;S~ N N
O O
,~ was prepared in an analogous fashion as described in example 1 starting from 4-methoxy-2,3,6-trimethylbenzenesulfonyl chloride and glycine methyl ester.
/
H O ~ / N
I' ~
,S; N v ' N N
~ i / O O ~ H H
OH 6_1 ~, was prepared in an analogous fashion as described in example 1 starting from a-toluenesulfonyl chloride and L-serine methyl ester.
H O ~ / N
I
~N~
;S, _ N N
~, was prepared in an analogous fashion as described in example 1 starting from a-toluenesulfonyl chloride and L-alanine methyl ester.
/
H O ~ / N
I
,S; N N N
/ ~ O CH3 H H 8-1 $~ was prepared in an analogous fashion as described in example 1 starting from a-toluenesulfonyl chloride and D-alanine methyl ester.
H O ~ / N
' ~ I
,S; N v 'N N
/ O O H H
~ was prepared in an analogous fashion as described in example 1 starting from a-toluenesulfonyl chloride and L-methionine methyl ester.
H O ~ / N
I' ~
,S; N v 'N N
" i O O H H
~ was prepared in an analogous fashion as described in example 1 starting from N-a-toluenesulfonyl-L-methionine sulfone methyl ester.
i H O ~ / N
I
~N~
~S~ N N
CI ~ O O H H
Stew l1:
SCI
o' ~o CI
A solution of 3,4-dichlorobenzyl chloride (4.3 mL, 31.1 mmol) in 15 mL of a 1:1 mixture of MeOH and water was treated with 4.9 g (31.1 mmol) of Na2S203 and the mixture was heated at 100°C for 3h.
After this time, the solution was cooled to ambient temperature and stirred for 16h. The resulting thick solid was diluted with 7 mL of HOAc and 7 mL of water to form a suspension. Chlorine gas was bubbled through the mixture for 20 min then the still heterogeneous mixture was stirred for 1h. The reaction mixture was diluted with 50 mL of ether and washed with 10 mL of 10% Na2S203, 10 mL of saturated NaHCOg, 10 mL of water and 10 mL of brine. The organic phase was dried over MgS04 and concentrated to afford the desired sulfonyl chloride ,~ as a white solid.
1H NMR (CDC13) 8 7.60 (d, J=2.2 Hz, 1H), 7.58 (d, J=?.8 Hz, 1H), 7.28 (dd, J=2.2, 7.8 Hz, 1H), 4.80 (s, 2H).
Step 2:
H O
i ,S~ Nv 'OH
O O
CI
Glycine methyl ester was prepared by the addition of 1.4 mL
(10 mmol) of Et3N to 1.25 g (IO mmol) of the amino ester HCl salt in 25 mL of DCM. After stirring for 10 min, 2.5 mL (30 mmol) of pyridine was added followed by 2.33 g (9.0 mmol) of 3,4-dichloro-a-toluenesulfonyl chloride ,1~- . The reaction mixture was stirred for 5 h, diluted with 100 mL of DCM then quenched with water. The organic phase was separated and washed with 5% Na2C03 (3 x 10 mL), 11,~ HCl (3 x 10 mL) and brine. Concentration of the organic phase afforded the crude ester (1.9 g, 6.1 mmol) which was dissolved in 30 mL of dioxane and 10 mL of water and treated with 1.28 g (30.5 mmol) of LiOH~H20. This new reaction mixture was stirred at room temperature for 17 h, concentrated and acidified to pH=1 with 1~ HCl. The solid was dissolved in EtOAc and the organic phase was separated and washed with brine. The solution was dried and concentrated to afford N-(3,4-dichlorobenzylsulfonyl)glycine ,],,~ as a white solid.
1H NMR (CDC13) 8 7.65 (s, 1H), 7.42 (d, J=7.8 Hz,1H), 7.38 (d, J=7.8 Hz, 1H), 7.02 (bt, 1H), 4.35 (s, 2H), 3.80 (d, J=6 Hz, 2H).
H O ~ / N
~N~
~S~ N N
CI I ~ O O H H
To a solution of the diamine ~ (385 mg, 1.7 mmol) in 5 mL
of DMF was added 506 mg (1.fi mmol) of the sulfonylated glycine ~ 229 mg (2.7 mmol) of HOBT, 32fi mg (1.7 mmol) of EDC and 0.60 mL (3.4 mmol) of DIPEA. The mixture was stirred to room temperature over I6 h, concentrated and chromatographed (5% MeOH in CHC13 saturated with NH3) to afford ,as a white solid.
1H NMR (DMSO-d6) b 8.05 (d, J=6.8 Hz, 2H), 7.1-7.6 (m, 8H), 6.40 (d, J=6.8 Hz, 2H), 5.25 (bt, 1H), 4.20 (s, 2H), 3.60 (s, 2H), 3.0-3.3 (m, 2H), 2.84 (t, 2H), 1.85 (m, 1H), 1.58 (m, 1H).
Anal. Calc'd for C24H26N4S03C12: C; 55.28, H; 5.03, N; 10.74. Found:
C; 55.58, H; 5.17, N; 10.68.
WO 99/16750 PC'T/US98/20174 H O ~ / N
~N~
;S~ N N
CI I / O O H H
~, was prepared in an analogous fashion as described in example 11 starting from 4-chlorobenzyl chloride.
/
H O ~ / N
,~ ~ I
;S, N N
O O H H
,~, was prepared in an analogous fashion as described in example 11 starting from 4-trifluoromethylbenzyl chloride.
o ~ ~ N
~N~
;S~ N N
F ~ ~ O O H H
was prepared in an analogous fashion as described in example 11 starting from 4-trifluoromethylbenzyl chloride.
In uitro assay for determin,'_n~ sroteinase i h; ition Assays of human a-thrombin and human trypsin were performed at 25°C in 0.05 M TRIS buffer pH 7.4, 0.15 M NaCl, 0.1% PEG.
Trypsin assays also contained 1 mM CaCl2.
In assays wherein rates of hydrolysis of a p-nitroanilide (pna) substrate were determined, a Thermomax 96-well plate reader was used to measure {at 405 nm) the time dependent appearance of p-nitroaniline. sar-PR-pna (sarcosine-Pro-Arg-p-nitroanilide) was used to assay human a-thrombin (Km=125 ~,tM) and human trypsin (Km=59 ~.1VI). p-Nitraanilide substrate concentration was determined from measurements of absorbance at 342 nm using an extinction coe~cient of 8270 cm-1M-1.
In certain studies with potent inhibitors (Ki < 10 nM) where the degree of inhibition of thrombin was high, a more sensitive activity assay was employed. In this assay the rate of thrombin catalyzed hydrolysis of the fluorogenic substrate Z-GPR-afc (Cbz-Gly-Pro-Arg-7-amino-4-trifluoromethyl coumarin) (Km=27 ~,M) was determined from the increase in fluorescence at 500 nm (excitation at 400 nm) associated with production of 7-amino-4-trifluoromethyl coumarin. Concentrations of stock solutions of Z-GPR-afc were determined from measurements of absorbance at 380 nm of the 7-amino-4-trifluoromethylcoumarin -3?-produced upon complete hydrolysis of an aliquot of the stock solution by thrombin.
Activity assays were performed by diluting a stock solution of substrate at least tenfold to a final concentration <_ 0.5 Km into a solution containing enzyme or enzyme equilibrated with inhibitor.
Times required to achieve equilibration between enzyme and inhibitor were determined in control experiments. Initial velocities of product formation in the absence (Vo) or presence of inhibitor (Vi) were measured. Assuming competitive inhibition, and that unity is negligible compared Km/(S], [I]/e, and [I]/e (where [S], [I], and a respectively represent the total concentrations, of substrate, inhibitor and enzyme), the equilibrium constant (Ki) for dissociation of the inhibitor from the enzyme can be obtained from the dependence of Vo/Vi on [I] shown in equation 1.
Vo/Vi = 1 + [I]/Ki (1) The activities shown by this assay indicate that the compounds of the invention are therapeutically useful for treating various conditions in patients suffering from unstable angina, refractory angina, myocardial infarction, transient ischemic attacks, atrial fibrillation, thrombotic stroke, embolic stroke, deep vein thrombosis, disseminated intravascular coagulation, and reocclusion or restenosis of recanalized vessels.
In Vavo Studies To Measure Thrombot» Occlusions Studies of 3-(2-Phenylethylamino)-6-methyl-1-(2-amino-6-methyl-5-methylenecarboxamidomethylpyridinyl)-2-pyridinone using the following rat ferric chloride assay substantially as described in ?'hrombosis Research, No. 60, page 269(1990) by Kurtz et al were used to determine in vivo activity of the thrombin inhibitors of the invention.
Male Sprague-Dawley rats (body weights 200-350 grams) were anesthetized with dial-urethane solution (0.1 ml/100 gm body weight i.p.), and a lateral tail vein was cannulated with a 23 gauge needle connected to a 12 inch length of PE50 tubing. The tubing was attached to a 3-way valve by a tubing adapter. Saline (control) or test compound, as appropriate, was administered via the tail vein catheter. A
tracheostomy was performed with a 0.75 inch length of PE205 tubing.
The right carotid artery is exposed and a 1.3 mm diameter Doppler flow probe was placed on the vessel. Body temperature was maintained at 37°C using a heat lamp.
6 rats were randomized to continuous intravenous infusions of saline or test compound administered via the tail vein. Test compound was administered at a rate of 10 ~.g/kg/min. Treatment infusions were initiated fi0 min before the placement of a 3 mm square piece of Whatman No. 1 filter paper saturated with 35% FeCl3 onto the exposed carotid artery distal to the llow probe. Treatment infusions were continued for an additional 90 minutes after the application of FeCl3 (total infusion duration 150 minutes) if thrombotic occlusions did not occur, or were terminated 30 minutes after thrombotic occlusion of the vessel. Time to occlusion was defined as the time from application of FeCl3 to thrombotic occlusion of the vessel. At the termination of the study (90 minutes after application of FeCl3 in animals which did not occlude, or at 30 minutes after thrombotic occlusion), 3 ml blood samples were drawn by cardiac puncture into 0.3 ml of 3.8% sodium citrate.
EKAMPL_F~
Tablet Preparation Tablets containing 25.0, 50.0, and 100.0 mg., respectively, of active compound ,~ is prepared as illustrated below:
TABLE FOR DOSES CONTAINING FROM
Amount-mg Active Compound ~, 25.0 50.0 100.0 Microcrystalline cellulose 37.25 100.0 200.0 Modified food corn starch 37.25 4.25 8.5 Magnesium stearate 0.50 0.75 1.5 All of the active compound, cellulose, and a portion of the corn starch are mixed and granulated to 10% corn starch paste. The resulting granulation is sieved, dried and blended with the remainder of the corn starch and the magnesium stearate. The resulting granulation is then compressed into tablets containing 25.0, 50.0, and 100.0 mg, respectively, of active ingredient per tablet.
Tablet Preparation Exemplary compositions of ,~ tablets are shown below:
Component 0.25 mg 2 mg 10 mg 50 mg Active ~ 0.500% 1.000% 5.000% 14.29%
mannitol 49.50% 49.25% 47.25% 42.61%
microcrystalline 49.50% 49.25% 47.25% 42.61%
cellulose magnesium stearate0.500% 0.500% 0.500% 0.500%
2, 10 and 50 mg tablets were film-coated with an aqueous dispersion of hydroxypropyl cellulose, hydroxypropyl methylcellulose and titanium dioxide, providing a nominal weight gain of 2.4%.
Tablet Drenaration via d,'_rect com~~reR~»n Active ~, mannitol and microcrystalline cellulose were sieved through mesh screens of specified size (generally 250 to 750 N,m) and combined in a suitable blender. The mixture was subsequently blended (typically 15 to 30 min) until the drug was uniformly distributed in the resulting dry powder blend. Magnesium stearate was screened and added to the blender, after which a precompression tablet blend was achieved upon additional mixing (typically 2 to 10 min). The precompression tablet blend was then compacted under an applied force, typically ranging from 0.5 to 2.5 metric tons, sufficient to yield tablets of suitable physical strength with acceptable disintegration times (specifications will vary with the size and potency of the compressed tablet). In the case of the 2, 10 and 50 mg potencies, the tablets were dedusted and film-coated with an aqueous dispersion of water-soluble polymers and pigment.
WO 99!16750 PCT/US98/20174 Tablet preparation via drv granulation Alternatively, a dry powder blend is compacted under modest forces and remilled to afford granules of specified particle size.
The granules are then mixed with magnesium stearate and tabletted as stated above.
Intravenous Formulations Intravenous formulations of ,~ were prepared according to general intravenous formulation procedures.
o~gonent Estimated range Active ,~ 0.12 - 0.61 mg D-glucuronic acid* 0.5 - 5 mg Mannitol NF 50-53 mg 1 N Sodium Hydroxide q.s. pH 3.9 - 4.1 Water for injection q.s. 1.0 mL
Exemplary compositions A-C are as follows:
Component 0. ~ ~
Active ~. 0.61 mg* 0.30** 0.15***
D-glucuronic acid* 1.94 mg 1.94 mg 1.94 mg Mannitol NF 51.2 mg 51.2 mg 51.2 mg 1 N Sodium. Hydroxide q.s. pH 4.0 q.s. pH 4.0 q.s. pH 4.0 Water for injectionq.s. 1.0 mL q.s. 1.0 mL q.s. 1.0 mL
* 0.50 mg free base ** 0.25 mg free base *** 0.12 mg free base Various other buffer acids, such as L-lactic acid, acetic acid, citric acid or any pharmaceutically acceptable acid/conjugate base with reasonable buffering capacity in the pH range acceptable for intravenous administration may be substituted for glucuronic acid.
_4.3_
/
H O ~ / N
I
,S; N N N
/ ~ O CH3 H H 8-1 $~ was prepared in an analogous fashion as described in example 1 starting from a-toluenesulfonyl chloride and D-alanine methyl ester.
H O ~ / N
' ~ I
,S; N v 'N N
/ O O H H
~ was prepared in an analogous fashion as described in example 1 starting from a-toluenesulfonyl chloride and L-methionine methyl ester.
H O ~ / N
I' ~
,S; N v 'N N
" i O O H H
~ was prepared in an analogous fashion as described in example 1 starting from N-a-toluenesulfonyl-L-methionine sulfone methyl ester.
i H O ~ / N
I
~N~
~S~ N N
CI ~ O O H H
Stew l1:
SCI
o' ~o CI
A solution of 3,4-dichlorobenzyl chloride (4.3 mL, 31.1 mmol) in 15 mL of a 1:1 mixture of MeOH and water was treated with 4.9 g (31.1 mmol) of Na2S203 and the mixture was heated at 100°C for 3h.
After this time, the solution was cooled to ambient temperature and stirred for 16h. The resulting thick solid was diluted with 7 mL of HOAc and 7 mL of water to form a suspension. Chlorine gas was bubbled through the mixture for 20 min then the still heterogeneous mixture was stirred for 1h. The reaction mixture was diluted with 50 mL of ether and washed with 10 mL of 10% Na2S203, 10 mL of saturated NaHCOg, 10 mL of water and 10 mL of brine. The organic phase was dried over MgS04 and concentrated to afford the desired sulfonyl chloride ,~ as a white solid.
1H NMR (CDC13) 8 7.60 (d, J=2.2 Hz, 1H), 7.58 (d, J=?.8 Hz, 1H), 7.28 (dd, J=2.2, 7.8 Hz, 1H), 4.80 (s, 2H).
Step 2:
H O
i ,S~ Nv 'OH
O O
CI
Glycine methyl ester was prepared by the addition of 1.4 mL
(10 mmol) of Et3N to 1.25 g (IO mmol) of the amino ester HCl salt in 25 mL of DCM. After stirring for 10 min, 2.5 mL (30 mmol) of pyridine was added followed by 2.33 g (9.0 mmol) of 3,4-dichloro-a-toluenesulfonyl chloride ,1~- . The reaction mixture was stirred for 5 h, diluted with 100 mL of DCM then quenched with water. The organic phase was separated and washed with 5% Na2C03 (3 x 10 mL), 11,~ HCl (3 x 10 mL) and brine. Concentration of the organic phase afforded the crude ester (1.9 g, 6.1 mmol) which was dissolved in 30 mL of dioxane and 10 mL of water and treated with 1.28 g (30.5 mmol) of LiOH~H20. This new reaction mixture was stirred at room temperature for 17 h, concentrated and acidified to pH=1 with 1~ HCl. The solid was dissolved in EtOAc and the organic phase was separated and washed with brine. The solution was dried and concentrated to afford N-(3,4-dichlorobenzylsulfonyl)glycine ,],,~ as a white solid.
1H NMR (CDC13) 8 7.65 (s, 1H), 7.42 (d, J=7.8 Hz,1H), 7.38 (d, J=7.8 Hz, 1H), 7.02 (bt, 1H), 4.35 (s, 2H), 3.80 (d, J=6 Hz, 2H).
H O ~ / N
~N~
~S~ N N
CI I ~ O O H H
To a solution of the diamine ~ (385 mg, 1.7 mmol) in 5 mL
of DMF was added 506 mg (1.fi mmol) of the sulfonylated glycine ~ 229 mg (2.7 mmol) of HOBT, 32fi mg (1.7 mmol) of EDC and 0.60 mL (3.4 mmol) of DIPEA. The mixture was stirred to room temperature over I6 h, concentrated and chromatographed (5% MeOH in CHC13 saturated with NH3) to afford ,as a white solid.
1H NMR (DMSO-d6) b 8.05 (d, J=6.8 Hz, 2H), 7.1-7.6 (m, 8H), 6.40 (d, J=6.8 Hz, 2H), 5.25 (bt, 1H), 4.20 (s, 2H), 3.60 (s, 2H), 3.0-3.3 (m, 2H), 2.84 (t, 2H), 1.85 (m, 1H), 1.58 (m, 1H).
Anal. Calc'd for C24H26N4S03C12: C; 55.28, H; 5.03, N; 10.74. Found:
C; 55.58, H; 5.17, N; 10.68.
WO 99/16750 PC'T/US98/20174 H O ~ / N
~N~
;S~ N N
CI I / O O H H
~, was prepared in an analogous fashion as described in example 11 starting from 4-chlorobenzyl chloride.
/
H O ~ / N
,~ ~ I
;S, N N
O O H H
,~, was prepared in an analogous fashion as described in example 11 starting from 4-trifluoromethylbenzyl chloride.
o ~ ~ N
~N~
;S~ N N
F ~ ~ O O H H
was prepared in an analogous fashion as described in example 11 starting from 4-trifluoromethylbenzyl chloride.
In uitro assay for determin,'_n~ sroteinase i h; ition Assays of human a-thrombin and human trypsin were performed at 25°C in 0.05 M TRIS buffer pH 7.4, 0.15 M NaCl, 0.1% PEG.
Trypsin assays also contained 1 mM CaCl2.
In assays wherein rates of hydrolysis of a p-nitroanilide (pna) substrate were determined, a Thermomax 96-well plate reader was used to measure {at 405 nm) the time dependent appearance of p-nitroaniline. sar-PR-pna (sarcosine-Pro-Arg-p-nitroanilide) was used to assay human a-thrombin (Km=125 ~,tM) and human trypsin (Km=59 ~.1VI). p-Nitraanilide substrate concentration was determined from measurements of absorbance at 342 nm using an extinction coe~cient of 8270 cm-1M-1.
In certain studies with potent inhibitors (Ki < 10 nM) where the degree of inhibition of thrombin was high, a more sensitive activity assay was employed. In this assay the rate of thrombin catalyzed hydrolysis of the fluorogenic substrate Z-GPR-afc (Cbz-Gly-Pro-Arg-7-amino-4-trifluoromethyl coumarin) (Km=27 ~,M) was determined from the increase in fluorescence at 500 nm (excitation at 400 nm) associated with production of 7-amino-4-trifluoromethyl coumarin. Concentrations of stock solutions of Z-GPR-afc were determined from measurements of absorbance at 380 nm of the 7-amino-4-trifluoromethylcoumarin -3?-produced upon complete hydrolysis of an aliquot of the stock solution by thrombin.
Activity assays were performed by diluting a stock solution of substrate at least tenfold to a final concentration <_ 0.5 Km into a solution containing enzyme or enzyme equilibrated with inhibitor.
Times required to achieve equilibration between enzyme and inhibitor were determined in control experiments. Initial velocities of product formation in the absence (Vo) or presence of inhibitor (Vi) were measured. Assuming competitive inhibition, and that unity is negligible compared Km/(S], [I]/e, and [I]/e (where [S], [I], and a respectively represent the total concentrations, of substrate, inhibitor and enzyme), the equilibrium constant (Ki) for dissociation of the inhibitor from the enzyme can be obtained from the dependence of Vo/Vi on [I] shown in equation 1.
Vo/Vi = 1 + [I]/Ki (1) The activities shown by this assay indicate that the compounds of the invention are therapeutically useful for treating various conditions in patients suffering from unstable angina, refractory angina, myocardial infarction, transient ischemic attacks, atrial fibrillation, thrombotic stroke, embolic stroke, deep vein thrombosis, disseminated intravascular coagulation, and reocclusion or restenosis of recanalized vessels.
In Vavo Studies To Measure Thrombot» Occlusions Studies of 3-(2-Phenylethylamino)-6-methyl-1-(2-amino-6-methyl-5-methylenecarboxamidomethylpyridinyl)-2-pyridinone using the following rat ferric chloride assay substantially as described in ?'hrombosis Research, No. 60, page 269(1990) by Kurtz et al were used to determine in vivo activity of the thrombin inhibitors of the invention.
Male Sprague-Dawley rats (body weights 200-350 grams) were anesthetized with dial-urethane solution (0.1 ml/100 gm body weight i.p.), and a lateral tail vein was cannulated with a 23 gauge needle connected to a 12 inch length of PE50 tubing. The tubing was attached to a 3-way valve by a tubing adapter. Saline (control) or test compound, as appropriate, was administered via the tail vein catheter. A
tracheostomy was performed with a 0.75 inch length of PE205 tubing.
The right carotid artery is exposed and a 1.3 mm diameter Doppler flow probe was placed on the vessel. Body temperature was maintained at 37°C using a heat lamp.
6 rats were randomized to continuous intravenous infusions of saline or test compound administered via the tail vein. Test compound was administered at a rate of 10 ~.g/kg/min. Treatment infusions were initiated fi0 min before the placement of a 3 mm square piece of Whatman No. 1 filter paper saturated with 35% FeCl3 onto the exposed carotid artery distal to the llow probe. Treatment infusions were continued for an additional 90 minutes after the application of FeCl3 (total infusion duration 150 minutes) if thrombotic occlusions did not occur, or were terminated 30 minutes after thrombotic occlusion of the vessel. Time to occlusion was defined as the time from application of FeCl3 to thrombotic occlusion of the vessel. At the termination of the study (90 minutes after application of FeCl3 in animals which did not occlude, or at 30 minutes after thrombotic occlusion), 3 ml blood samples were drawn by cardiac puncture into 0.3 ml of 3.8% sodium citrate.
EKAMPL_F~
Tablet Preparation Tablets containing 25.0, 50.0, and 100.0 mg., respectively, of active compound ,~ is prepared as illustrated below:
TABLE FOR DOSES CONTAINING FROM
Amount-mg Active Compound ~, 25.0 50.0 100.0 Microcrystalline cellulose 37.25 100.0 200.0 Modified food corn starch 37.25 4.25 8.5 Magnesium stearate 0.50 0.75 1.5 All of the active compound, cellulose, and a portion of the corn starch are mixed and granulated to 10% corn starch paste. The resulting granulation is sieved, dried and blended with the remainder of the corn starch and the magnesium stearate. The resulting granulation is then compressed into tablets containing 25.0, 50.0, and 100.0 mg, respectively, of active ingredient per tablet.
Tablet Preparation Exemplary compositions of ,~ tablets are shown below:
Component 0.25 mg 2 mg 10 mg 50 mg Active ~ 0.500% 1.000% 5.000% 14.29%
mannitol 49.50% 49.25% 47.25% 42.61%
microcrystalline 49.50% 49.25% 47.25% 42.61%
cellulose magnesium stearate0.500% 0.500% 0.500% 0.500%
2, 10 and 50 mg tablets were film-coated with an aqueous dispersion of hydroxypropyl cellulose, hydroxypropyl methylcellulose and titanium dioxide, providing a nominal weight gain of 2.4%.
Tablet Drenaration via d,'_rect com~~reR~»n Active ~, mannitol and microcrystalline cellulose were sieved through mesh screens of specified size (generally 250 to 750 N,m) and combined in a suitable blender. The mixture was subsequently blended (typically 15 to 30 min) until the drug was uniformly distributed in the resulting dry powder blend. Magnesium stearate was screened and added to the blender, after which a precompression tablet blend was achieved upon additional mixing (typically 2 to 10 min). The precompression tablet blend was then compacted under an applied force, typically ranging from 0.5 to 2.5 metric tons, sufficient to yield tablets of suitable physical strength with acceptable disintegration times (specifications will vary with the size and potency of the compressed tablet). In the case of the 2, 10 and 50 mg potencies, the tablets were dedusted and film-coated with an aqueous dispersion of water-soluble polymers and pigment.
WO 99!16750 PCT/US98/20174 Tablet preparation via drv granulation Alternatively, a dry powder blend is compacted under modest forces and remilled to afford granules of specified particle size.
The granules are then mixed with magnesium stearate and tabletted as stated above.
Intravenous Formulations Intravenous formulations of ,~ were prepared according to general intravenous formulation procedures.
o~gonent Estimated range Active ,~ 0.12 - 0.61 mg D-glucuronic acid* 0.5 - 5 mg Mannitol NF 50-53 mg 1 N Sodium Hydroxide q.s. pH 3.9 - 4.1 Water for injection q.s. 1.0 mL
Exemplary compositions A-C are as follows:
Component 0. ~ ~
Active ~. 0.61 mg* 0.30** 0.15***
D-glucuronic acid* 1.94 mg 1.94 mg 1.94 mg Mannitol NF 51.2 mg 51.2 mg 51.2 mg 1 N Sodium. Hydroxide q.s. pH 4.0 q.s. pH 4.0 q.s. pH 4.0 Water for injectionq.s. 1.0 mL q.s. 1.0 mL q.s. 1.0 mL
* 0.50 mg free base ** 0.25 mg free base *** 0.12 mg free base Various other buffer acids, such as L-lactic acid, acetic acid, citric acid or any pharmaceutically acceptable acid/conjugate base with reasonable buffering capacity in the pH range acceptable for intravenous administration may be substituted for glucuronic acid.
_4.3_
Claims (32)
1. A compound having the following structure:
and pharmaceutically acceptable salts thereof, wherein n is 0 or 1;
R is hydrogen C1-6 alkyl, arylC0-6alkylene-, C3-8cycloalkyl, or -CH2C3-8cycloalkyl;
R1 is hydrogen, C1-6alkyl hydroxy, C1-6alkyl, -(CH2)m SO2R3, wherein m is 0-6, arylC0-6alkylene, wherein aryl is unsubstituted or mono-, di-, tri-, or tetra- substituted with C1-4 alkyl, hydroxy, C1-4alkoxy, or CF3;
-(CH2)mSR3, wherein m is 0-6, or C3-8cycloalkyl;
R2 is Boc, -SO2R4, -SO2-CH=CHR4, hydrogen, C1-6alkyl, C3-8cycloalkyl, R4, -C(O)R5, or -SO2R5;
R3 is C1-6alkyl; or R4 is arylC0-6alkylene, wherein aryl is unsubstituted or mono-, di-, tri-, or tetra- substituted with C1-4 alkyl, hydroxy, C1-4alkoxy, halogen, amino, or CF3;
R5 is hydrogen, C1-6alkyl, C3-8cycloalkyl, or arylC0-6alkyl.
and pharmaceutically acceptable salts thereof, wherein n is 0 or 1;
R is hydrogen C1-6 alkyl, arylC0-6alkylene-, C3-8cycloalkyl, or -CH2C3-8cycloalkyl;
R1 is hydrogen, C1-6alkyl hydroxy, C1-6alkyl, -(CH2)m SO2R3, wherein m is 0-6, arylC0-6alkylene, wherein aryl is unsubstituted or mono-, di-, tri-, or tetra- substituted with C1-4 alkyl, hydroxy, C1-4alkoxy, or CF3;
-(CH2)mSR3, wherein m is 0-6, or C3-8cycloalkyl;
R2 is Boc, -SO2R4, -SO2-CH=CHR4, hydrogen, C1-6alkyl, C3-8cycloalkyl, R4, -C(O)R5, or -SO2R5;
R3 is C1-6alkyl; or R4 is arylC0-6alkylene, wherein aryl is unsubstituted or mono-, di-, tri-, or tetra- substituted with C1-4 alkyl, hydroxy, C1-4alkoxy, halogen, amino, or CF3;
R5 is hydrogen, C1-6alkyl, C3-8cycloalkyl, or arylC0-6alkyl.
2. A compound of Claim 1, having the structure and pharmaceutically acceptable salts thereof, wherein R is
3. A compound of Claim 2, and pharmaceutically acceptable salts thereof, wherein R1 is hydrogen, -CH2OH, CH3, -CH2CH2SCH3, -CH2CH2SO2CH3.
4. A compound of Claim 3, and pharmaceutically acceptable salts thereof, wherein R2 is hydrogen,
5. A compound of Claim 4, and pharmaceutically acceptable salts thereof, selected from the group consisting of
6. A composition comprising a compound of Claim 1 and a pharmaceutically acceptable carrier.
7. A method for inhibiting thrombin in blood comprising adding to the blood a composition of Claim 6.
8. A method for inhibiting formation of blood platelet aggregates in blood comprising adding to the blood a composition of Claim 6.
9. A method for inhibiting thrombus formation in blood comprising adding to the blood a composition of Claim 6.
10. A method for inhibiting thrombus formation in blood comprising adding to the blood a compound of Claim 1 with a fibrinogen receptor antagonist.
11. The use of a compound of Claim 1, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for inhibiting thrombus formation, preventing thrombus formation, inhibiting thrombin, inhibiting formation of fibrin, and inhibiting formation of blood platelet aggregates, in a mammal.
12. A method for treating or preventing venous thromboembolism and pulmonary embolism in a mammal comprising administering to the mammal a composition of Claim 6.
13. A method for treating or preventing deep vein thrombosis in a mammal comprising administering to the mammal a composition of Claim 6.
14. A method for treating or preventing cardiogenic thromboembolism in a mammal comprising administering to the mammal a composition of Claim 6.
15. A method for treating or preventing thromboembolic stroke in humans and other mammals comprising administering to the mammal a composition of Claim 6.
16. A method for treating or preventing thrombosis associated with cancer and cancer chemotherapy in a mammal comprising administering to the mammal a composition of Claim 6.
17. A method for treating or preventing unstable angina in a mammal comprising administering to the mammal a composition of Claim 6.
18. A method for treating or preventing myocardial infarction in a mammal comprising administering to the mammal a composition of Claim 6.
19. A method for treating or preventing cardiogenic thromboembolism associated with atrial fibrillation in a mammal comprising administering to the mammal a composition of Claim 6.
20. A method for treating or preventing cardiogenic thromboembolism associated with prosthetic heart valves in a mammal comprising administering to the mammal a composition of Claim 6.
21. A method for treating or preventing cardiogenic thromboembolism associated with heart disease in a mammal comprising administering to the mammal a composition of Claim 6.
22. A method for treating or preventing atherosclerosis in a mammal comprising administering to the mammal a composition of Claim 6.
23. A method for treating or preventing thrombosis in a mammal with inherited thrombophilic diseases such as Protein C
deficiency, Protein S deficiency, antithrombin III and factor V Leiden comprising administering to the mammal a composition of Claim 6.
deficiency, Protein S deficiency, antithrombin III and factor V Leiden comprising administering to the mammal a composition of Claim 6.
24. A method for treating or preventing thrombosis in a mammal with acquired thrombophilic disorders such as systemic lupus erythematosus comprising administering to the mammal a composition of Claim 6.
25. A method for treating or preventing ischemic heart disease in a mammal comprising administering to the mammal a composition of Claim 6.
26. A method for lowering the propensity of a device which contacts blood to clot blood which comprises coating the device with a composition of claim 6.
27. A method for treating or preventing reocclusion in a mammal during or following percutaneous transluminal coronary angioplasty comprising administering to the mammal a composition of Claim 6.
28. A method for treating or preventing restenosis in a mammal following percutaneous transluminal coronary angioplasty comprising administering to the mammal a composition of Claim 6.
29. A method for treating or preventing occlusion of coronary artery bypass grafts in a mammal comprising administering to the mammal a composition of Claim 6.
30. A method for treating or preventing occlusive cerebrovascular disease in a mammal comprising administering to the mammal a composition of Claim 6.
31. A method for maintaining patency in arteriovenous cannulas inserted in a mammal comprising administering to the mammal a composition of Claim 6.
32. A method for maintaining patency of indwelling catheters in a mammal comprising administering to the mammal a composition of Claim 6.
Applications Claiming Priority (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US6053697P | 1997-09-30 | 1997-09-30 | |
| US60/060,536 | 1997-09-30 | ||
| GB9806034.6 | 1998-03-20 | ||
| GBGB9806034.6A GB9806034D0 (en) | 1998-03-20 | 1998-03-20 | Thrombin inhibitors |
| PCT/US1998/020174 WO1999016750A1 (en) | 1997-09-30 | 1998-09-25 | Thrombin inhibitors |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2303832A1 true CA2303832A1 (en) | 1999-04-08 |
Family
ID=26313327
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002303832A Abandoned CA2303832A1 (en) | 1997-09-30 | 1998-09-25 | Thrombin inhibitors |
Country Status (5)
| Country | Link |
|---|---|
| EP (1) | EP1027333A4 (en) |
| JP (1) | JP2001518466A (en) |
| AU (1) | AU740170B2 (en) |
| CA (1) | CA2303832A1 (en) |
| WO (1) | WO1999016750A1 (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE19956786A1 (en) * | 1999-11-25 | 2001-05-31 | Basf Ag | New optically active aminoalcohol and diamine derivatives useful as intermediates, e.g. for pharmaceuticals and plant protectants |
| US7857522B2 (en) * | 2007-01-31 | 2010-12-28 | Siemens Industry, Inc. | Rolling mill oil film bearing |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| ES2051702T3 (en) * | 1986-04-01 | 1994-07-01 | Wako Pure Chem Ind Ltd | PEPTIDIC DERIVATIVES AND METHOD FOR MEASURING THE ACTIVITY OF PHYSIOLOGICALLY ACTIVE SUBSTANCES USING THEM AS SUBSTRATES. |
| DE4306873A1 (en) * | 1993-03-05 | 1994-09-08 | Boehringer Mannheim Gmbh | New 4-aminopyridine processes for their preparation and medicaments containing these compounds |
| US5668289A (en) * | 1996-06-24 | 1997-09-16 | Merck & Co., Inc. | Pyridinone thrombin inhibitors |
-
1998
- 1998-09-25 EP EP98949536A patent/EP1027333A4/en not_active Withdrawn
- 1998-09-25 WO PCT/US1998/020174 patent/WO1999016750A1/en not_active Ceased
- 1998-09-25 JP JP2000513836A patent/JP2001518466A/en not_active Withdrawn
- 1998-09-25 AU AU95839/98A patent/AU740170B2/en not_active Ceased
- 1998-09-25 CA CA002303832A patent/CA2303832A1/en not_active Abandoned
Also Published As
| Publication number | Publication date |
|---|---|
| AU740170B2 (en) | 2001-11-01 |
| AU9583998A (en) | 1999-04-23 |
| EP1027333A1 (en) | 2000-08-16 |
| JP2001518466A (en) | 2001-10-16 |
| EP1027333A4 (en) | 2000-11-08 |
| WO1999016750A1 (en) | 1999-04-08 |
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