Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
  • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
  • C07D405/12—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a chain containing hetero atoms as chain links
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P25/00—Drugs for disorders of the nervous system
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P25/00—Drugs for disorders of the nervous system
  • A61P25/22—Anxiolytics
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P25/00—Drugs for disorders of the nervous system
  • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P27/00—Drugs for disorders of the senses
  • A61P27/02—Ophthalmic agents
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P27/00—Drugs for disorders of the senses
  • A61P27/02—Ophthalmic agents
  • A61P27/06—Antiglaucoma agents or miotics
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P5/00—Drugs for disorders of the endocrine system
  • A61P5/38—Drugs for disorders of the endocrine system of the suprarenal hormones
  • A61P5/42—Drugs for disorders of the endocrine system of the suprarenal hormones for decreasing, blocking or antagonising the activity of mineralocorticosteroids
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P9/00—Drugs for disorders of the cardiovascular system
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P9/00—Drugs for disorders of the cardiovascular system
  • A61P9/04—Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P9/00—Drugs for disorders of the cardiovascular system
  • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P9/00—Drugs for disorders of the cardiovascular system
  • A61P9/12—Antihypertensives

Definitions

• ACE inhibitors and ATi blockers have been accepted as treatments of hypertension (Waeber B. et al., "The renin-angiotensin system: role in experimental and human hypertension,” in Berkenhager W. H., Reid J. L. (eds): Hypertension, Amsterdam, Elsevier Science Publishing Co, 1996, 489-519; Weber M. A., Am. J. Hypertens., 1992, 5, 247S).
• ACE inhibitors are used for renal protection (Rosenberg M. E.
• renin inhibitors stems from the specificity of renin (Kleinert H. D., Cardiovasc. Drugs, 1995, 9, 645).
• the only substrate known for renin is angiotensinogen, which can only be processed (under physiological conditions) by renin.
• ACE can also cleave bradykinin besides Ang I and can be bypassed by chymase, a serine protease (Husain A., J. Hypertens., 1993, 11, 1155).
• ACE can also cleave bradykinin besides Ang I and can be bypassed by chymase, a serine protease (Husain A., J. Hypertens., 1993, 11, 1155).
• inhibition of ACE thus leads to bradykinin accumulation, causing cough (5-20%) and potentially life-threatening angioneurotic edema (0.1- 0.2%) (Konili Z. H.
• R 1 is alkyl, cycloalkyl or cycloalkylalkyl
• an aspartic protease inhibitor which is a compound represented by Structural Formula (III):
• E for each occurrence, is independently H or an amine protecting group.
• Amine protecting groups include carbamate, amide, and sulfonamide protecting groups known in the art (T. W. Greene and P. G. M. Wuts "Protective Groups in Organic Synthesis” John Wiley & Sons, Inc., New York 1999, the entire teaching of which is herein incorporated by reference).
• Specific amine protecting groups include tert- butoxycarbonyl (Boc), benzyloxycarbonyl (Cbz) and 1 -[2- (trimethylsilyl)ethoxycarbonyl] (Teoc). Values and specific values for R 2 are as described for Structural Formula (I).
• the intermediate is each of the following compounds or their enantiomers or diastereomers.
• Pharmaceutically acceptable salts of all of the following are also included:
• Racemate or “racemic mixture” means a compound of equimolar quantities of two enantiomers, wherein such mixtures exhibit no optical activity; i.e., they do not rotate the plane of polarized light.
• R and S indicate configurations relative to the core molecule.
• Conventional resolution techniques include forming the salt of a free base of each isomer of an isomeric pair using an optically active acid (followed by fractional crystallization and regeneration of the free base), forming the salt of the acid form of each isomer of an isomeric pair using an optically active amine (followed by fractional crystallization and regeneration of the free acid), forming an ester or amide of each of the isomers of an isomeric pair using an optically pure acid, amine or alcohol (followed by chromatographic separation and removal of the chiral auxiliary), or resolving an isomeric mixture of either a starting material or a final product using various well known chromatographic methods.
• a disclosed aspartic protease inhibitor is named or depicted by structure without indicating the stereochemistry, and the inhibitor has at least one chiral center, it is to be understood that the name or structure encompasses one enantiomer of inhibitor free from the corresponding optical isomer, a racemic mixture of the inhibitor and mixtures enriched in one enantiomer relative to its corresponding optical isomer.
• Salts of the compounds of the aspartic protease inhibitors containing an acidic functional group can be prepared by reacting with a suitable base.
• a suitable base which affords a pharmaceutically acceptable cation, which includes alkali metal salts (especially sodium and potassium), alkaline earth metal salts (especially calcium and magnesium), aluminum salts and ammonium salts, as well as salts made from physiologically acceptable organic bases such as trimethylamine, triethylamine, morpholine, pyridine, piperidine, picoline, dicyclohexylamine, N 5 N'- dibenzylethylenediamine, 2-hydroxyethyl amine, bis-(2-hydroxyethyl)amine, tri-(2- hydroxyethyl)amine, procaine, dibenzylpiperidine, dehydroabietylamine, N 5 N'- bisdehydroabietylamine, glucamine, N-methylglucamine, collidine, quinine, quinoline
• a disclosed aspartic protease inhibitor When a disclosed aspartic protease inhibitor is named or depicted by structure, it is to be understood that the compound, including solvates thereof, may exist in crystalline forms, non-crystalline forms or a mixture thereof. The aspartic protease inhibitor or solvates may also exhibit polymorphism (i.e. the capacity to occur in different crystalline forms). These different crystalline forms are typically known as "polymorphs.” It is to be understood that when named or depicted by structure, the disclosed aspartic protease inhibitors and solvates (e.g., hydrates) also include all polymorphs thereof. Polymorphs have the same chemical composition but differ in packing, geometrical arrangement, and other descriptive properties of the crystalline solid state.
• a pharmaceutical composition of the invention may, alternatively or in addition to a disclosed aspartic protease inhibitor, comprise a prodrug or pharmaceutically active metabolite of such a compound or salt and one or more pharmaceutically acceptable carriers or diluent therefor.
• the invention includes a therapeutic method for treating or ameliorating an aspartic protease mediated disorder in a subject in need thereof comprising adrainistering to a subject in need thereof an effective amount of an aspartic protease inhibitor disclosed herein.
• Effective amount means that amount of drug substance (i.e. aspartic protease inhibitors of the present invention) that elicits the desired biological response in a subject. Such response includes alleviation of the symptoms of the disease or disorder being treated.
• the effective amount of a disclosed aspartic protease inhibitor in such a therapeutic method is from about .01 mg/kg/day to about 10 mg/kg/day, preferably from about 0.5 mg/kg/day to 5 mg/kg/day.
• “Pharmaceutically acceptable carrier” means compounds and compositions that are of sufficient purity and quality for use in the formulation of a composition of the invention that, when appropriately administered to an animal or human, do not produce an adverse reaction, and that are used as a vehicle for a drug substance (i.e. aspartic protease inhibitors of the present invention).
• “”Pharmaceutically acceptable diluent” means compounds and compositions that are of sufficient purity and quality for use in the formulation of a composition of the invention that, when appropriately administered to an animal or human, do not produce an adverse reaction, and that are used as a diluting agent for a drug substance (i.e. aspartic protease inhibitors of the present invention).
• Aspartic protease mediated disorder or disease includes disorders or diseases associated with the elevated expression or overexpression of aspartic proteases and conditions that accompany such diseases.
• An embodiment of the invention includes administering an aspartic protease inhibitor disclosed herein in a combination therapy (see USP 5,821,232, USP 6,716,875, USP 5,663,188, Fossa, A. A.; DePasquale, M. J.; Ringer, L. J.; Winslow, R. L.
• a diuretic is, for example, a thiazide derivative selected from amiloride, chlorothiazide, hydrochlorothiazide, methylchlorothiazide, and chlorothalidon.
• Centrally acting antiphypertensives include clonidine, guanabenz, guanfacine and methyldopa.
• ACE inhibitors include alacepril, benazepril, benazaprilat, captopril, ceronapril, cilazapril, delapril, enalapril, enalaprilat, fosinopril, lisinopril, moexipiril, moveltopril, perindopril, quinapril, quinaprilat, ramipril, ramiprilat, spirapril, temocapril, trandolapril, and zofenopril.
• Preferred ACE inhibitors are benazepril, enalpril, lisinopril, and ramipril.
• Preferred HIV integrase inhibitors are L-870,810 and S-1360.
• Entry inhibitors include compounds that bind to the CD4 receptor, the CCR5 receptor or the CXCR4 receptor.
• Specific examples of entry inhibitors include enfuvirtide (a peptidomimetic of the HR2 domain in gp41) and sifurvitide.
• a preferred attachment and fusion inhibitor is enfuvirtide.
• An embodiment of the invention includes administering an aspartic protease inhibitor disclosed herein or composition thereof in a combination therapy with one or more additional agents for the treatment of Alzheimer's disease including tacrine, donepezil, rivastigmine, galantamine, and memantine.
• An aspartic protease inhibitor disclosed herein may also be prepared by grinding, micronizing or other particle size reduction methods known in the art. Such methods include, but are not limited to, those described in U.S. Pat. Nos. 4,826,689, 5,145,684, 5,298,262, 5,302,401, 5,336,507, 5,340,564, 5,346,702, 5,352,459, 5,354,560, 5,384,124, 5,429,824, 5,503,723, 5,510,118, 5,518,187, 5,518,738, 5,534,270, 5,536,508, 5,552,160, 5,560,931, 5,560,932, 5,565,188, 5,569,448, 5,571,536, 5,573,783, 5,580,579, 5,585,108, 5,587,143, 5,591,456, 5,622,938, 5,662,883, 5,665,331, 5,718,919, 5,747,001, PCT applications WO 93/25190, WO 96/24336, and WO
• compositions of the invention include ocular, oral, nasal, transdermal, topical with or without occlusion, intravenous (both bolus and infusion), and injection (intraperitoneally, subcutaneously, intramuscularly, intratumorally, or parenterally).
• compositions of the invention suitable for oral administration include solid forms such as pills, tablets, caplets, capsules (each including immediate release, timed release, and sustained release formulations), granules and powders; and, liquid forms such as solutions, syrups, elixirs, emulsions, and suspensions.
• forms useful for ocular administration include sterile solutions or ocular delivery devices.
• forms useful for parenteral administration include sterile solutions, emulsions, and suspensions.
• the oral composition is preferably formulated as a homogeneous composition, wherein the drug substance (i.e. aspartic protease inhibitors of the present invention) is dispersed evenly throughout the mixture, which may be readily subdivided into dosage units containing equal amounts of a disclosed aspartic protease inhibitor.
• drug substance i.e. aspartic protease inhibitors of the present invention
• compositions are prepared by mixing a disclosed aspartic protease inhibitor with one or more optionally present pharmaceutical carriers (such as a starch, sugar, diluent, granulating agent, lubricant, glidant, binding agent, and disintegrating agent), one or more optionally present inert pharmaceutical excipients (such as water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents, and syrup), one or more optionally present conventional tableting ingredients (such as corn starch, lactose, sucrose, sorbitol, talc, stearic acid, magnesium stearate, dicalcium phosphate, and any of a variety of gums), and an optional diluent (such as water).
• pharmaceutical carriers such as a starch, sugar, diluent, granulating agent, lubricant, glidant, binding agent, and disintegrating agent
• inert pharmaceutical excipients such as water, glycols, oils, alcohols, flavoring agents
• Biodegradable and non-biodegradable slow release carriers are well known in the art.
• Biodegradable carriers are used to form particles or matrices which retain drug substance(s) (i.e. aspartic protease inhibitors of the present invention) and which slowly degrade/dissolve in a suitable environment (e.g., aqueous, acidic, basic and the like) to release drug substances.
• a suitable environment e.g., aqueous, acidic, basic and the like
• Such particles degrade/dissolve in body fluids to release the drug substance(s) (i.e. aspartic protease inhibitors of the present invention) therein.
• an ophthalmic composition may contain one or more of: a) a surfactant such as a polyoxyethylene fatty acid ester; b) a thickening agents such as cellulose, cellulose derivatives, carboxyvinyl polymers, polyvinyl polymers, and polyvinylpyrrolidones, typically at a concentration n the range of about 0.05 to about 5.0% (wt/vol); c) (as an alternative to or in addition to storing the composition in a container containing nitrogen and optionally including a free oxygen absorber such as Fe), an anti-oxidant such as butylated hydroxyanisol, ascorbic acid, sodium thiosulfate, or butylated hydroxytoluene at a concentration of about 0.00005 to about 0.1% (wt/vol); d) ethanol at a concentration of about 0.01 to 0.5% (wt/vol); and e) other excipients such as an isotonic agent
• Representative compounds of the invention can be synthesized in accordance with the general synthetic schemes described above and are illustrated in the examples that follow. The methods for preparing the various starting materials used in the schemes and examples are well within the knowledge of persons skilled in the art.
• (R)-tert-buty ⁇ 3-((i?)-(2-aminoethoxy)(3- chlorophenyl)methyl)-piperidine-l -carboxylate may also be prepared by the following procedures:
• HPLC analysis showed 88% conversion with minor impurities as well as approx. 9% starting alcohol.
• the reaction was allowed to cool to RT and the layers separate. The addition of 10 vol. of water was needed to ensure the clean separation of the layers.
• the organic layer was retained and rinsed with 10 vol brine. The organic layer was retained and concentrated under vacuum.
• the resulting residual oil was dissolved in 10 vol tert-buty ⁇ methyl ether (TBME) at which point 10 vol of a 20% weight solution of citric acid in water was added. (Note: tartaric acid works as well while acids such as HCl, oxalic acid, TsOH result in deprotection of the NBoc).
• HPLC analysis showed that clean extraction of the desired amine into the aq.
• the aq. layer was rinsed once more with 5 vol of TBME in order to ensure the removal of the undesired starting alcohol.
• the organic TBME layer was discarded.
• the aq. layer was brought to a pH of approx. 13 by the addition of 2 vol of 50% weight NaOH in water at which point 10 vol
• the Grignard reagent was added dropwise to a solution of ( ⁇ )-tert- butyl 3-(methoxy(methyl)carbarnoyl)piperidine-l-carboxylate (27.2 g, 0.1 mol) in anhydrous THF (300 mL) at -78 0 C under nitrogen. After addition, the mixture was allowed to stir at rt for 1.5 hr. The mixture was quenched with saturated NH 4 Cl solution (300 mL) and extracted with EtOAc (3x200 mL).
• Step 8 (R)-tert-butyl 3-(( ⁇ )-(2-azidoethoxy)(5-chloro-2- methylphenyl)methyl)piperidine- 1 -carboxylate
• Step 8 (S)-tert-butyl 4-((i2)-5-te ⁇ butoxy-5-oxo-2-(tosyloxymethyl)pentyl)-2,2- dimethyloxazolidine-3 -carboxy late
• Step 6 dibenzyl l-((25)-l-hydroxy-3-(tetrahydro-2//-pyran-3-yl)propan-2- yl)hydrazine- 1 ,2-dicarboxylate
• Step 1 tert-buty ⁇ (iS)-l-hydroxy-3-(( ⁇ )-tetrahydro-2H-pyran-3-yl)propan-2- ylcarbamate (5)-/er/-Butyl-2,2-dimethyl-4-(((i?)-tetrahydro-2//-pyran-3- yl)methyl)oxazolidine- 3-carboxylate (9 g, 30.1 mmol) was dissolved in 80% aq CH 3 CO 2 H (90ml). The solution was stirred at 50 0 C during 1.5 hr and evaporated to dryness at reduced pressure. The residue was dissolved in Et 2 O (150ml) and washed with saturated NaHCO 3 (4 ⁇ 100 mL) .
• Step 1 methyl 2-((/?)-(3-chlorophenyl)(( ⁇ )-piperidin-3-yl)methoxy)ethylcarbamate
• Step 1 (4-nitrophenyl) ( ⁇ S)-l-(N-methyl-N-(trimethylsilylethoxycarbonyl)amino)-3- (( ⁇ )-tetrahydro -2H-py ran-3 -yl)propan-2-y lcarbamate
• Step 3 methyl 2-((R)-Q -chloro-5-fluorophenylXCR)- 1 -((S)- 1 -(methylamino)-3- ((/f)-tetrahydro-2H-pyran-3-yl)propan-2-ylcarbamoyl)piperidin-3- yl)methoxy)ethylcarbamate
• Step l methyl 2-(( ⁇ )-(3-fluorophenyl)((3i?)-l-((-S)-l-(N-Methyl-2-
• Step 3 methyl 2-((/e)-(3-fluorophenyl)(( ⁇ )-l-((6)-l-(methylamino)-3-(( ⁇ )- tetrahydro-2H-pyran-3 ⁇ yl)propan-2-ylcarbamoyl)piperidin-3- yl)methoxy)ethylcarbamate and methyl 2-(( ⁇ )-(3-fluorophenyl)(( ⁇ )-l -((S)-I - (methylamino)-3-((-?)-tetrahydro-2H-pyran-3-yI)propan-2-ylcarbamoyl)piperidin-3- yl)methoxy)ethylcarbamate
• Step 4 methyl 2-((i.)-(3-fluorophenyl)((i-)-l-((5)-l-(methylamino)-3-((i?)- tetrahy dro-2H-pyran-3 ⁇ yl)propan-2-ylcarbamoyl)piperidin-3 - yl)methoxy)ethylcarbamate fumaric acid salt
• Step 5 methyl 2-(( ⁇ )-(3-fluorophenyl)((/?)-l-((5)-l-(methylamino)-3-((5)- tetrahy dro-2H-pyran-3 -y l)propan-2-ylcarbamoyl)piperidin-3 - yl)methoxy)ethylcarbamate fumaric acid salt
• Step 1 methyl 2-((i?)-(( ⁇ )-l-((5)-l-(methylamino)-3-(( ⁇ )-tetrahydro-2//-pyran-3- yl)propan-2-ylca ⁇ bamoyl)piperidin-3-yl)(phenyl)methoxy)ethylcarbamate
• Step 1 methyl 2-((i.)-(3-chlorophenyl)((/-)-l-((5)-l-(methylamino)-3-((i?)- tetrahydro-2//-pyran-3-yl)propan-2-ylcarbamoyl)piperidin-3- yl)methoxy)ethylcarbamate
• Step 2 methyl 2-((22)-(3-chlorophenyl)(CK)-l -((S)A -(methylamino)-3 -((£)- tetrahydro-2/f-pyran-3-yl)propan-2-ylcarbamoyl)piperidin-3- yl)methoxy)ethylcarbamate fumaric acid salt
• aspartic protease inhibitors of the invention When the stereochemistry at a chiral center is not defined in the compound name, this indicates that the sample prepared contained a mixture of isomers at this center.
• the disclosed aspartic protease inhibitors have enzyme-inhibiting properties. In particular, they inhibit the action of the natural enzyme renin.
• the latter passes from the kidneys into the blood where it effects the cleavage of angiotensinogen, releasing the decapeptide angiotensin I which is then cleaved in the blood, lungs, the kidneys and other organs by angiotensin converting enzyme to form the octapeptide angiotensin II.
• the octapeptide increases blood pressure both directly by binding to its receptor, causing arterial vasoconstriction, and indirectly by liberating from the adrenal glands the sodium-ion-retaining hormone aldosterone, accompanied by an increase in extracellular fluid volume.
• That increase can be attributed to the action of angiotensin II.
• Inhibitors of the enzymatic activity of renin bring about a reduction in the formation of angiotensin I. As a result a smaller amount of angiotensin II is produced.
• the reduced concentration of that active peptide hormone is the direct cause of the hypotensive effect of renin inhibitors.
• renin inhibitors The action of renin inhibitors in vitro was demonstrated experimentally by means of a test which measures the increase in fluorescence of an internally quenched peptide substrate.
• the sequence of this peptide corresponds to the sequence of human angiotensinogen.
• the following test protocol was used. All reactions were carried out in a flat bottom white opaque microtiter plate.
• trypsin-activated recombinant human renin final enzyme concentration of 0.2-2 nM
• the increase in fluorescence at 495 nm (excitation at 340 nm) is measured for 60-360 minutes at rt using a Perkin-Elmer Fusion microplate reader.
• the slope of a linear portion of the plot of fluorescence-increase as a function of time was then determined, and the rate was used for calculating percent inhibition in relation to uninhibited control.
• the percent inhibition values were plotted as a function of inhibitor concentration, and the IC 50 is determined from a fit of this data to a four parameter equation.
• the IC 50 was defined as the concentration of a particular inhibitor that reduces the formation of product by 50% relative to a control sample containing no inhibitor.
• the disclosed aspartic protease inhibitors exhibit inhibiting activities at minimum concentrations of from approximately 5 x 10 "5 M to approximately 10 "12 M.
• Specific aspartic protease inhibitors exhibit inhibiting activities at minimum concentrations of from approximately lO '7 M to approximately 10 "12 M.
• renin inhibitors in vitro in human plasma were demonstrated experimentally by the decrease in plasma renin activity (PRA) levels observed in the presence of the compounds.
• PRA plasma renin activity
• Incubations mixtures contained in the final volume of 250 ⁇ L 95.5 mM N,N-bis(2-hydroxyethyl)-2-aminoethanesulfonic acid, pH 7.0, 8 mM EDTA, 0.1 mM neomycin sulfate, 1 mg/mL sodium azide, 1 mM phenylmethanesulfonyl fluoride, 2% DMSO and 87.3% of pooled mixed-gender human plasma stabilized with EDTA.
• cardiac and systemic hemodynamic efficacy of selective renin inhibitors can be evaluated in vivo in sodium-depleted, normotensive cynomolgus monkeys. Arterial blood pressure is monitored by telemetry in freely moving, conscious animals.
• Cynomolgus Monkey (prophetic example): Six male naive cynomolgus monkeys weighing between 2.5 and 3.5 kg are to be used in the studies. At least 4 weeks before the experiment, the monkeys are anesthetized with ketamine hydrochloride (15 mg/kg, i.m.) and xylazine hydrochloride (0.7 mg/kg, i.m.), and are implanted into the abdominal cavity with a transmitter (Model #TL11M2-D70-PCT, Data Sciences, St. Paul, MN)- The pressure catheter is inserted into the lower abdominal aorta via the femoral artery. The bipotential leads are placed in Lead II configuration.
• the animals are housed under constant temperature (19-25°C), humidity (>40%) and lighting conditions (12 h light and dark cycle), are fed once daily, and are allowed free access to water.
• the animals are sodium depleted by placing them on a low sodium diet (0.026%, Expanded Primate Diet 829552 MP- VENaCl (P), Special Diet Services, Ltd., UK) 7 days before the experiment and furosemide (3 mg/kg, intramuscularly i.m., Aventis Pharmaceuticals) is administered at -40 h and -16 h prior to administration of test compound.
• the renin inhibitors are formulated in 0.5% methylcellulose at dose levels of 10 and 30 mg/kg (5 mL/kg) by infant feeding tubes.
• a silastic catheter is implanted into posterior vena cava via a femoral vein. The catheter is attached to the delivery pump via a tether system and a swivel joint.
• Test compound dose levels of 0.1 to 10 mg/kg, formulated at 5% dextrose
• Double Transgenic Rat The efficacy of the renin inhibitor 6a was evaluated in vivo in double transgenic rats engineered to express human renin and human angiotensinogen (Bohlender J 5 Fukamizu A, Lippoldt A, Nomura T, Dietz R 5 Menard J 5 Murakami K, Vietnamese FC, Ganten D. High human renin hypertension in transgenic rats. Hypertension 1997, 29, 428-434). Experiments were conducted in 6- week-old double transgenic rats (dTGRs).
• the human renin construct used to generate transgenic animals made up the entire genomic human renin gene (10 exons and 9 introns), with 3.0 IcB of the 5'-promoter region and 1.2 kB of 3 1 additional sequences.
• the human angiotensinogen construct made up the entire human angiotensinogen gene (5 exons and 4 introns), with 1.3 kB of 5'-flanking and 2.4 kB of 3'-flanking sequences.
• the rats were purchased from RCC Ltd (F ⁇ llinsdorf, Switzerland). Radio telemetry transmitters were surgically implanted at 4 weeks of age.
• the telemetry system provided 24-h recordings of systolic, mean, diastolic arterial pressure (SAP, MAP, DAP, respectively) and heart rate (HR). Beginning on day 42, animals were transferred to telemetry cages. A 24 h telemetry reading was obtained. Rats were then dosed orally on the following 4 consecutive days (days 43-46). The rats were monitored continuously and allowed free access to standard 0.3%-sodium rat chow and drinking water.

Landscapes

• Health & Medical Sciences (AREA)
• Chemical & Material Sciences (AREA)
• Organic Chemistry (AREA)
• Engineering & Computer Science (AREA)
• Bioinformatics & Cheminformatics (AREA)
• General Health & Medical Sciences (AREA)
• Veterinary Medicine (AREA)
• General Chemical & Material Sciences (AREA)
• Medicinal Chemistry (AREA)
• Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
• Chemical Kinetics & Catalysis (AREA)
• Pharmacology & Pharmacy (AREA)
• Life Sciences & Earth Sciences (AREA)
• Animal Behavior & Ethology (AREA)
• Public Health (AREA)
• Cardiology (AREA)
• Neurology (AREA)
• Neurosurgery (AREA)
• Heart & Thoracic Surgery (AREA)
• Biomedical Technology (AREA)
• Ophthalmology & Optometry (AREA)
• Hospice & Palliative Care (AREA)
• Diabetes (AREA)
• Endocrinology (AREA)
• Urology & Nephrology (AREA)
• Vascular Medicine (AREA)
• Psychiatry (AREA)
• Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
• Plural Heterocyclic Compounds (AREA)
• Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Priority Applications (4)

Applications Claiming Priority (2)

Publications (2)

Family

ID=38926325

Family Applications (1)

Country Status (5)

Cited By (8)

Families Citing this family (6)

Citations (2)

Family Cites Families (18)

• 2007
  • 2007-09-17 JP JP2009528319A patent/JP2010503682A/ja not_active Withdrawn
  • 2007-09-17 AT AT07838310T patent/ATE520690T1/de not_active IP Right Cessation
  • 2007-09-17 EP EP07838310A patent/EP2081927B1/en active Active
  • 2007-09-17 WO PCT/US2007/020086 patent/WO2008036216A1/en not_active Ceased
  • 2007-09-17 US US12/311,020 patent/US20090318501A1/en not_active Abandoned

Patent Citations (2)

Also Published As

Similar Documents

Legal Events