WO1993006127A1 - Novel amino acid prodrug renin inhibitors - Google Patents

Novel amino acid prodrug renin inhibitors Download PDF

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Publication number
WO1993006127A1
WO1993006127A1 PCT/US1992/007463 US9207463W WO9306127A1 WO 1993006127 A1 WO1993006127 A1 WO 1993006127A1 US 9207463 W US9207463 W US 9207463W WO 9306127 A1 WO9306127 A1 WO 9306127A1
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Prior art keywords
cad
phe
smo
defined above
atm
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PCT/US1992/007463
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French (fr)
Inventor
Xue-Min Cheng
Joseph Thomas Repine
Michael Douglas Taylor
Jonathan Leonard Wright
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Warner-Lambert Company
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Publication of WO1993006127A1 publication Critical patent/WO1993006127A1/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D295/00—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms
    • C07D295/22—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with hetero atoms directly attached to ring nitrogen atoms
    • C07D295/26—Sulfur atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/02—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link
    • C07K5/0215—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link containing natural amino acids, forming a peptide bond via their side chain functional group, e.g. epsilon-Lys, gamma-Glu
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06008—Dipeptides with the first amino acid being neutral
    • C07K5/06078—Dipeptides with the first amino acid being neutral and aromatic or cycloaliphatic
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06191—Dipeptides containing heteroatoms different from O, S, or N
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • the present invention relates to novel amino acid prodrug renin inhibitors useful as
  • novel compounds of the present invention inhibit the enzyme renin thus controlling hypertension
  • hyperaldosteronism congestive heart failure
  • glaucoma as well as the use of the compounds as diagnostic tools in mammals.
  • Renin is a natural enzyme which is released into the blood stream from the kidney. It cleaves its natural substrate, angiotensinogen, releasing a decapeptide, angiotensin I. This in turn is cleaved by converting enzyme in the lung, kidney, and other tissues to an octapeptide, angiotensin II.
  • Angiotensin II raises blood pressure both directly by causing arteriolar constriction and indirectly by stimulating release of the sodium-retaining hormone aldosterone from the adrenal gland causing a rise in extracellular fluid volume.
  • Inhibitors of renin have been sought as agents for control of hypertension and hyperaldosteronism.
  • HIV protease like renin, is an aspartyl protease
  • novel compounds of the present invention also can be used to treat disorders caused by retroviruses including HTLV I, II, and III.
  • Renin inhibitors have been shown to be effective agents in the control of hypertension and related cardiovascular diseases. However, in general they suffer the drawbacks of poor aqueous solubility and bioavailability. Thus, there is a need to develop a renin inhibitor that has good aqueous solubility and bioavailability.
  • the present invention is a compound of Formula I
  • R is hydrogen or alkyl of from one to six carbon atoms, wherein R is as defined above,
  • R 3 is 2
  • R 4 is hydrogen or hydroxyl
  • X is O, S, or NH
  • R 2 is alkyl of from one to six carbon atoms ; - wherein R 5 is R 5
  • R 6 , R 7 , R 8 , or R 9 are each independently hydrogen, alkyl of from one to six carbon atoms, alkoxy of from one to six carbon atoms, halogen, or trifluoromethyl.
  • R 1 and X are as defined above,
  • R 1 is as defined above
  • R 1 and X are as defined above, or wherein R 1 and X are as defined above;
  • R 5 is as defined above, or
  • alkyl of from one to six carbon atoms
  • n and R 1 are as defined above,
  • R 1 is as defined above
  • R 1 , X, R 5 , and R 10 are as defined above;
  • R 12 is
  • R 1 and X are as defined above, wherein R 1 and X are as defined above,
  • R 14 is or
  • R 11 and p are as defined above;
  • R 1 with the exclusion of R 1 being hydrogen is encompassed within the definition of at least one of A, E, G, or J; or a pharmaceutically acceptable salt thereof.
  • amino acid prodrug renin inhibitors the compounds of Formula I are useful for the treatment of hypertension, hyperaldosteronism, congestive heart failure, and diseases caused by retroviruses
  • HTLC I, II, and III as well as diagnostic tools for the identification of cases of hypertension due to renin excess.
  • the present invention is directed to methods for production of a compound of Formula I.
  • alkyl means a straight or branched hydrocarbon radical having from one to six carbon atoms and includes, for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, n-hexyl, and the like.
  • Alkoxy is O-alkyl as defined above for alkyl.
  • Halogen is fluorine, chlorine, bromine, or iodine.
  • TsOH Para-toluenesulfonic acid An amino acid or organic acid within the bracket in a structure means the bracketed moiety is attached via the ⁇ carboxylic acid or carboxylic acid,
  • an ester such as, for example:
  • bracketed amino acid or organic acid forms an amide bond with the amine such as, for example:
  • prodrug refers to a compound of Formula I which is biotransformed into the active renin inhibitor in a mammal.
  • the compounds of Formula I are capable of further forming pharmaceutically acceptable acid addition and/or base salts. All of these forms are within the scope of the present invention.
  • Pharmaceutically acceptable acid addition salts of the compound of Formula I include salts derived from nontoxic inorganic acids such as hydrochloric, nitric, phosphoric, sulfuric, hydrobromic, hydriodic, phosphorous and the like, as well as the salts derived from nontoxic organic acids such as aliphatic mono- and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxy alkanoic acids, alkanedioic acids, aromatic acids, aliphatic and aromatic
  • Such salts thus include
  • dihydrogenphosphate metaphosphate, pyrophosphate. chloride, bromide, iodide, acetate, propionate, caprylate, isobutyrate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, mandelate, benzoate, chlorobenzoate, methylbenzoate,
  • toluenesulfonate phenylacetate, citrate, lactate, maleate, tartrate, methanesulfonate and the like.
  • salts of amino acids such as arginate and the like and gluconate, galacturonate (see, for example. Bergs S. M., et al,
  • the acid addition salts of said basic compounds are prepared by contacting the free base form with a sufficient amount of the desired acid to produce the salt in the conventional manner.
  • the free base form may be regenerated by contacting the salt form with a base and isolating the free base in the conventional manner.
  • the free base forms differ from their respective salt forms somewhat in certain physical properties such as solubility in polar solvents, but otherwise the salts are equivalent to their
  • Pharmaceutically acceptable base addition salts of acidic compounds are formed with metals or amines such as alkali and alkaline earth metals or organic amines.
  • metals used as cations are sodium, potassium, magnesium, calcium and the like.
  • suitable amines are sodium, potassium, magnesium, calcium and the like.
  • N-methylglucamine N-methylglucamine
  • procaine see, for example, Berge S. M., et al. Journal of Pharmaceutical
  • the base addition salts of said acidic compounds are prepared by contacting the free acid form with a sufficient amount of the desired base to produce the salt in the conventional manner.
  • the free acid form may be regenerated by contacting the salt form with an acid and isolating the free acid in the
  • the free acid forms differ from their respective salt forms somewhat in certain physical properties such as solubility in polar solvents, but otherwise the salts are equivalent to their respective free acids for purposes of the present invention.
  • inventions can exist in unsolvated forms as well as solvated forms, including hydrated forms.
  • solvated forms including hydrated forms
  • unsolvated forms are equivalent to unsolvated forms and are intended to be encompassed within the scope of the present invention.
  • the compounds of the present invention possess one or more chiral centers and each center may exist in the R(D) or S(L) configuration.
  • the present invention includes all enantiomeric forms as well as the appropriate mixtures thereof.
  • a preferred compound of Formula I is one wherein A is
  • R is hydrogen or alkyl of from one to six carbon atoms
  • R 2 is alkyl of from one to six carbon atoms
  • R 6 , R 7 , R 8 , or R 9 are each independently hydrogen, alkyl of from one to six carbon atoms, alkoxyl of from one to six carbon atoms, halogen or trifluoromethyl,
  • R 6 , R 7 , R 8 , or R 9 are each independently hydrogen, alkyl of from one to six carbon atoms, alkoxy of from one to six carbon atoms, halogen, or trifluoromethyl.
  • R 1 and X are as defined above.
  • R 1 is as defined above.
  • alkyl of from one to six carbon atoms , -CO 2 CH 3 ,
  • n is zero or an integer of 1 or 2 and R 1 is as defined above, -(CH 2 ) n -CONH 2 wherein n is as defined above,
  • R 1 is as defined above
  • R 1 , X, R 5 , and R 10 are as defined above;
  • R 12 is wherein R 1 is as defined above. wherein R 1 is as defined
  • R 14 is
  • R 1 with the exclusion of R 1 being hydrogen is encompassed within the definition of at least one of A, E, G, or J.
  • a more preferred compound of Formula I is one wherein J is
  • R 11 is alkyl
  • R 12 is 2
  • R 1 is hydrogen, - wherein R 3 is
  • n is as defined above, and X is O, S, or NH, or wherein R 14 is
  • R 11 is as defined above.
  • Stability in Intestinal Perfusate - Perfusate is generated by oscillating 15 to 20 mL MES buffer through rat jejunum in situ at 30 mL/min using a withdrawal/infusion pump (Harvard Apparatus,
  • prodrug collected and spiked with prodrug at 37 ⁇ g/mL.
  • the solution is incubated at 37°C and disappearance of prodrug over time is monitored by direct injection (25 ⁇ L) onto HPLC.
  • Membranes (BBM) - BBM are prepared from rat or rabbit small intestine by the method of Kessler, et al,
  • Vesicles 50 ⁇ L are combined with 450 ⁇ L drug or prodrug (10 ⁇ g/mL) and incubated at 37°C. At selected time points, 100 ⁇ L prodrug/BBM suspension is removed and diluted with 100 ⁇ L
  • HPLC Conditions - Samples are analyzed by high pressure liquid chromatography using a two pump binary-gradient system (HP 1090 Liquid
  • MES buffer is 10 mM (2-[N-Morpholino]ethanesulfonic acid, made iso-osmotic with 10 mM KCL and 140 mM NaCl and adjusted to pH 6.5 with IN NaOH.
  • Table I show some representative water soluble amino acid prodrug renin inhibitors which are converted to the parent renin inhibitor by intestinal enzymes and also by brush border enzymes.
  • R is hydrogen or alkyl of from one to six carbon atoms. -N- - wherein R is as defined above,
  • n is zero or an integer of 1 or 2 and R 4 is hydrogen or
  • X is O, S, or NH
  • R 2 is alkyl of from one to six carbon atoms
  • R 6 , R 7 , R 8 , or R 9 are each independently hydrogen, alkyl of from one to six carbon atoms, alkoxy of from one to six carbon atoms, halogen, or
  • R 1 is as defined above
  • G is N wherein R 5 is as defined above, or
  • alkyl of from one to six carbon atoms
  • n and R 1 are as defined above,
  • R 1 is as defined above
  • R 1 , X, R 5 , and R 10 are as defined above; J is wherein R 11 is
  • R 12 is 3
  • R 1 and X are as defined above, wherein R 1 and X are as defined above,
  • R 14 is or
  • R 11 and p are as defined above;
  • R 1 with the exclusion of R 1 being hydrogen is encompassed within the definition of at least one of A, E, G, or J; or a pharmaceutically acceptable salt thereof comprises:
  • A', E', G', and J' are as defined above for A, E, G, and J provided R 1 is hydrogen and is encompassed within the definition of at least one of A', E', G', or J' with a compound of Formula III
  • R 1a is as defined above for R 1 but excluding R 1 is hydrogen and providing any basic or acidic groups contain conventional protecting groups and X is as defined above to afford a compound of Formula IV
  • A'', E'', G'', and J'' are as defined above for A, E, G, and J provided R 1a is as defined above and is encompassed within the definition of at least one of A", E'' , G" or J";
  • a compound of Formula II may be prepared by sequential stepwise or fragment coupling of the amino acids or fragments selected from A', E', G', or J' to the preceding amino acid or fragment using
  • a compound of Formula IV is prepared from anino acids or fragments selected from A'', E'', G" , or J'' using the methodology used to prepare a compound of Formula II.
  • a compound of Formula III is either known or capable of being prepared by methods known in the art.
  • Compounds of Formula I are designated by numbers 8, 11, 14, 24a, 24b, 30a, 30b, 40, 43, 54, an 58.
  • the compound of Formula 4 is treated with hydrogen in the presence of a catalyst such as, for example, 5% to 20% palladium on carbon, platinum oxide, 5% to 20% platinum on carbon, and the like in a solvent such as, for example, EtOH, EtOAc, THF, dioxane, DMF and the like at about 0°C to about 100°C for about 30 minutes to about 24 hours to give the compound of Formula 5.
  • a catalyst such as, for example, 5% to 20% palladium on carbon, platinum oxide, 5% to 20% platinum on carbon, and the like in a solvent such as, for example, EtOH, EtOAc, THF, dioxane, DMF and the like at about 0°C to about 100°C for about 30 minutes to about 24 hours to give the compound of Formula 5.
  • the compound of Formula 2 is reacted with the compound of Formula 5 in the presence of a coupling reagent such as, for example, DCC and HOBT and the like and a solvent such as, for example, CHCl 3 , CH 2 Cl 2 , THF, dioxane, EtOAc, DMF, DMSO and the like at about 0°C to about 40°C for about 1 to 5 days to give the compound of Formula 6.
  • a coupling reagent such as, for example, DCC and HOBT and the like
  • a solvent such as, for example, CHCl 3 , CH 2 Cl 2 , THF, dioxane, EtOAc, DMF, DMSO and the like at about 0°C to about 40°C for about 1 to 5 days to give the compound of Formula 6.
  • the compound of Formula 6 is reacted with LYS.2BOC in the presence of a coupling reagent such as, for example, CDI, DCC, and the like in the presence of a solvent such as, for example, CHCl 3 , CH 2 Cl 2 , THF, dioxane, EtOAc, DMF, DMSO and the like at about 0°C to about 100°C for about 3 hours to about 3 days to give the compound of Formula 7.
  • the compound of Formula 7 is treated with an acid such as, for example, trifluoroacetic acid, or a nonpolar inert solvent such as CH 2 Cl 2 saturated with hydrogen chloride gas at about -20°C to about 25°C for about
  • a compound of Formula 10 and a compound of Formula 13 are reacted with hydrogen in the presence of a catalyst such as for example 50% palladium on carbon and an acid such as for example TFA and the like in a solvent such as for example ethanol and the like toafford, respectively, a compound of Formula 11 and a compound of Formula 14.
  • a catalyst such as for example 50% palladium on carbon and an acid such as for example TFA and the like in a solvent such as for example ethanol and the like toafford, respectively, a compound of Formula 11 and a compound of Formula 14.
  • the compound of Formula 16 is treated with a hydride reagent such as, for example, KBH 4 and the like in a solvent such as, for example, absolute EtOH and the like to give the compound of Formula 17 as a mixture of R and S isomers at the carbon atom towhich the hydroxyl group is attached.
  • Choice of solvent and reducing reagent can vary the ratio of isomers obtained to give a predominant excess of a particular desired isomer.
  • the R and S isomers of the compound of Formula 17 may be separated into the individual R and S isomers by conventional separation techniques such as, for example, chromatography, fractional crystallization and the like.
  • R and S isomers of the compound of Formula 18 may be separated into the individual R and S isomers by conventional separation techniques such as, for example, chromatography and the like. Additionally, other hydroxyl protecting groups such as, for
  • acetyl, benzyl, and the like may be used in place of the TBDMS group with subsequent separation of the R and S protected hydroxyl groups following the methodology used to separate the isomers of the compound of Formula 18.
  • Either the compound of Formula 18a (R-isomer) or Formula 18b (S-isomer) is reacted with a fluoride ion source such as, for example, t-butylammonium fluoride and the like in a solvent such as, for example, THF and the like to give the compound of Formula 19a (R-isomer) or Formula 19b (S-isomer).
  • a fluoride ion source such as, for example, t-butylammonium fluoride and the like
  • solvent such as, for example, THF and the like
  • N-protecting groups other than Boc or Cbz may also be used to protect the amino groups of Lys
  • a solvent such as, for example, CH 2 Cl 2 and the like at about ambient temperature to give the compound of Formula 20a (R-isomer) or
  • Formula 20a (R-isomer) or Formula 20b (S-isomer) is treated with an oxidizing reagent such as, for example, NaIO 4 with a catalytic amount of RuO 2 , and the like to give the compound of Formula 21a (R- isomer) or Formula 21b (S-isomer).
  • an oxidizing reagent such as, for example, NaIO 4 with a catalytic amount of RuO 2 , and the like to give the compound of Formula 21a (R- isomer) or Formula 21b (S-isomer).
  • Either the compound of Formula 21a (R-isomer) or Formula 21b (S- isomer) is reacted with CAD in the presence of a coupling reagent such as, for example, DCC and HOBT and the like in a solvent such as, for example,
  • aprotic solvent such as, for example, DMF, DMSO and the like is added to aid solution, at about 0°C to about 25°C for about 4 hours to about 3 days to give the compound of Formula 22a (R-isomer) or Formula 22b (S-isomer). Either the compound of Formula 22a
  • Formula 23a (R-isomer) or Formula 23b (S-isomer) is treated with an acid such as, for example,
  • Formula 24b (S-isomer) may be converted to other pharmaceutically acceptable acid addition salts by conventional methodology as previously described.
  • Formula 19b is reacted with 2,2-dimethoxypropane in the presence of an acid catalyst such as, for
  • a compound of Formula I designated as 40 is prepared from a compound of Formula 31 as outlined in Scheme 5.
  • a compound of Formula I designated as 43 is prepared from a compound of Formula 32 as outlined in Scheme 6.
  • inhibitors of compounds of Formula I designated as 65a, 65b, 71, and 73 are as outlined in Schemes 8 and 9.
  • the compounds of the present invention can be prepared and administered in a wide variety of oral and parenteral dosage forms. It will be obvious to those skilled in the art that the following dosage forms may comprise as the active component, either a compound of Formula I or a corresponding
  • pharmaceutically acceptable carriers can be either solid or liquid.
  • Solid form preparations include powders, tablets, pills, capsules, cachets,
  • a solid carrier can be one or more substances which may also act as diluents, flavoring agents, binders,
  • preservatives for example, preservatives, tablet disintegrating agents, or an encapsulating material.
  • the carrier is a finely divided solid which is in a mixture with the finely divided active component.
  • the active component is mixed with the carrier having the necessary binding properties in suitable proportions and compacted in the shape and size desired.
  • the powders and tablets preferably contain from five or ten to about seventy percent of the active compound.
  • Suitable carriers are magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth,
  • preparation is intended to include the formulation of the active compound with encapsulating material as a carrier providing a capsule in which the active component with or without other carriers, is
  • Tablets powders, capsules, pills,
  • cachets, and lozenges can be used as solid dosage forms suitable for oral administration.
  • a low melting wax such as a mixture of fatty acid glycerides or cocoa butter
  • the active component is dispersed homogeneously therein, as by stirring.
  • the molten homogenous mixture is then poured into
  • Liquid form preparations include solutions, suspensions, and emulsions, for example, water or water propylene glycol solutions.
  • liquid preparations can be formulated in solution in aqueous polyethylene glycol solution.
  • Aqueous solutions suitable for oral use can be prepared by dissolving the active component in water and adding suitable colorants, flavors, stabilizing and thickening agents as desired.
  • Aqueous suspensions suitable for oral use can be made by dispersing the finely divided active
  • viscous material such as natural or synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose, and other well-known suspending agents.
  • solid form preparations which are intended to be converted, shortly before use, to liquid form preparations for oral administration.
  • Such liquid foirms include solutions, suspensions, and emulsions. These preparations may contain, in addition to the active component, colorants, flavors, stabilizers, buffers, artificial and natural
  • the pharmaceutical preparation is preferably in unit dosage form. In such form the preparation is subdivided into unit doses containing appropriate quantities of the active component.
  • the unit dosage form can be a packaged preparation, the package containing discrete quantities of preparation, such as packeted tablets, capsules, and powders in vials or ampoules. Also, the unit dosage form can be a capsules, tablet, cachet, or lozenge itself, or it can be the appropriate number of any of these in packaged form.
  • the quantity of active component in a unit dose preparation may be varied or adjusted from 0.1 mg to 2000 mg preferably 0.5 mg to 1000 mg according to the particular application and the potency of the active component.
  • the composition can, if desired, also contain other compatible therapeutic agents.
  • the compounds utilized in the pharmaceutical method of this invention are HTLV I, II, and III as well as diagnostic tools for the identification of cases of hypertension due to renin excess, the compounds utilized in the pharmaceutical method of this invention are
  • the dosages may be varied depending upon the requirements of the patient, the severity of the condition being treated, and the compound being employed. Determination of the proper dosage for a particular situation is within the skill of the art. Generally, treatment is initiated with smaller dosages which are less than the optimum dose of the compound. Thereafter, the dosage is increased by small increments until the optimum effect under the circumstances is reached. For convenience, the total daily dosage may be divided and administered in portions during the day, if desired.
  • Step A Preparation of SMO-Phe-Ser (Lys•2 Boc)-CAD Di-t-Boc-Lys (1.11 g, 3.19 mmol, 2.0 eq) in THF
  • Step B Preparation of SMO-Phe-Ser(Asp•HCl)-CAD
  • Hydrogen chloride gas is passed over the
  • Step A Preparation of SMO-Phe-Thr(Lys•2Boc)-CAD
  • Example D (1.926 g, 3.00 mmol) in THF (20 mL) and DMAP (100 mg) are added at 0°C. The mixture is stirred at room temperature overnight and then at reflux for 24 hours. The cooled mixture is
  • Step B Preparation of SMO-Phe-Thr(Lys•2HCl)-CAD
  • Step A Preparation of SMO-Phe-Ser(Glu•Cbz)-CAD
  • Step B Preparation of SMO-Phe-Ser(Glu•TFA)-CAD
  • Step A Preparation of SMO-Phe-Ser(Gln•Cbz)-CAD
  • Step B Preparation of SMO-Phe-Ser(Gln•TFA)-CAD
  • Step A Preparation of SMO-Phe-Ser(Ala-Cbz)-CAD
  • Step A Preparation of (Lys-2Boc)O-CH 2 -Phe-Alg-CAD
  • Step B Preparation of (Lys•2HCl)O-CH 2 -Phe-Alg-CAD
  • Step B Preparation of (Asp•HCl)O-CH, -Phe-Alg- CAD•HCl
  • Step A Preparation of (Asp•Boc•t-Bu) OCH 2 -Phe-Atm-CAD
  • Example U is dissolved in 25 mL of MeOH and 200 mg of TsOH (3 eq) and a catalytic amount of 20% palladium on carbon is added. The mixture is stirred under one atmosphere of hydrogen for 6 hours, filtered, and the solution neutralized with solid NaHCO 3 . The MeOH is evaporated and the residue diluted with water and EtOAc. The mixture is extracted with EtOAc (3 x 30 mL) and dried (magnesium sulfate). The product is purified by chromatography with 3% MeOH:CH 2 Cl 2 to afford 140 mg of product as a white foam; MS (FAB) 932 (M +1 ).
  • Step B Preparation of -Phe-Atm-CAD•HCl
  • Step B Preparation of (Asp)O 2 -CAD 3
  • Boc-Alg-CAD (Example K) (1.01 g, 2.29 mmol) is dissolved in 100 mL of dry CH 2 Cl 2 . Hydrogen chloride gas is passed through this solution for 10 minutes. The reaction mixture is stirred for 3 hours. Solvent is removed and the residue dissolved in H 2 O (50 mL). The aqueous solution is extracted with Et 2 O
  • Boc-Phe-Alg-CAD (Example M) (7.0 g, 11.92 mmol) is dissolved in 500 mL of CH 2 Cl 2 with the aid of a few drops of MeOH. Dry HCl gas is passed through solution for 15 minutes and the reaction mixture is further stirred for another 3 hours. Solvent is evaporated and the residue dissolved in H 2 O (200 mL). It is then extracted with CH 2 Cl 2 (3 x 250 mL). The aqueous layer is basified with NaOH pellets and extracted with CH 2 Cl 2 (3 x 300 mL). The combined extracts are dried (MgSO 4 ) and evaporated to give 5.70 g of white solid; MS (FAB) 489 (M + +1) 471
  • Step A Preparation of (Lys-2Boc) O-CH 2 -CO 2 Bz
  • Carbonyldiimidazole (2.25 g, 13.88 mmol) is added to the solution of Bis-Boc Lysine (4.00 g, 11.56 mmol) in 40 mL of THF at 0°C followed by a catalytic amount of DMAP. After 10 minutes, benzyl glycolate (1.28 g, 7.7 mmol) is added and the
  • Example H is dissolved in 140 mL acetone and cooled to 15°C.
  • NaIO 4 9.50 g (44.41 mmol) and RuO 2 xH 2 O,
  • N-Boc•Asp•O-t-Bu (17.1 g, 53 mmol), freed from the dicyclohexylamine salt by treatment with citric acid is dissolved in 300 mL of THF and
  • Patent Application EP0399,556 6.3 g (14.9 mmol), CAD, 4.0 g (1.1 eq), DCC, 3.4 g (1.1 eq), HOBT, 2.2 g (1.1 eq), in 50 mL of DMF is stirred at room temperature
  • Boc-Atm(Cbz)-CAD 2.5 g (3.86 mmol) is dissolved in 40 mL of CH 2 Cl 2 and 3 mL of MeOH to afford a clear solution. Hydrogen chloride gas is passed through the solution for 10 minutes, and the reaction stirred for another 3.5 hours. The solvent is removed and the residue dissolved in 100 mL of EtOAc and 100 mL of NaHCO 3 . The layers are separated and the EtOAc layer is washed with NaCl and dried (magnesium sulfate). The solvent is evaporated to afford 2.2 g of product as a pale white foam which is used without further purification.

Abstract

Novel amino acid prodrug renin inhibitors are described, as well as methods for the preparation and pharmaceutical compositions of the same, which are useful as renin inhibitors and thus useful in controlling hypertension, hyperaldosteronism, congestive heart failure, and glaucoma, as well as diagnostic agents.

Description

NOVEL AMINO ACID PRODRUG RENIN INHIBITORS
BACKGROUND OF THE INVENTION
The present invention relates to novel amino acid prodrug renin inhibitors useful as
pharmaceutical agents, to methods for their
production, to pharmaceutical compositions which include these compounds and a pharmaceutically acceptable carrier, and to pharmaceutical methods of treatment as well as the use of these agents as diagnostic tools. More particularly, the novel compounds of the present invention inhibit the enzyme renin thus controlling hypertension,
hyperaldosteronism, congestive heart failure, and glaucoma as well as the use of the compounds as diagnostic tools in mammals.
Renin is a natural enzyme which is released into the blood stream from the kidney. It cleaves its natural substrate, angiotensinogen, releasing a decapeptide, angiotensin I. This in turn is cleaved by converting enzyme in the lung, kidney, and other tissues to an octapeptide, angiotensin II.
Angiotensin II raises blood pressure both directly by causing arteriolar constriction and indirectly by stimulating release of the sodium-retaining hormone aldosterone from the adrenal gland causing a rise in extracellular fluid volume. Inhibitors of renin have been sought as agents for control of hypertension and hyperaldosteronism.
Additionally, since HIV protease, like renin, is an aspartyl protease, the novel compounds of the present invention also can be used to treat disorders caused by retroviruses including HTLV I, II, and III.
Renin inhibitors have been shown to be effective agents in the control of hypertension and related cardiovascular diseases. However, in general they suffer the drawbacks of poor aqueous solubility and bioavailability. Thus, there is a need to develop a renin inhibitor that has good aqueous solubility and bioavailability.
We have found unexpectedly that certain amino acid derivatives of renin inhibitors have increased water solubility and bioavailability. These
derivatives are stable in the gastrointestinal tract but undergo selective, enzyme catalyzed cleavage of the amino acid auxiliary at the gut wall to produce a high local concentration of the parent renin
inhibitor and hence improved transport across the gut wall. This effect, combined with the much higher gut lumen concentration of compound due to increased solubility, greatly enhances oral adsorption and hence overall bioavailability. Some of the parent compounds have been disclosed in United States Patent U.S. 5,036,053, European Published Application
EP 0399,556, and Repine, J. T., et al. Journal of Medicinal Chemistry, Volume 34, pages 1935-1943
(1991) as renin inhibitors.
SUMMARY OF THE INVENTION
Accordingly, the present invention is a compound of Formula I
A-E-G-J I wherein A is
wherein R is hydrogen or alkyl of from one to six carbon atoms,
Figure imgf000005_0001
wherein R is as defined above,
Figure imgf000005_0002
wherein R is as defined above.
Figure imgf000005_0003
Figure imgf000005_0004
or
Figure imgf000006_0001
Figure imgf000006_0002
wherein R1 is
hydrogen,
wherein R3 is
Figure imgf000006_0003
2
hydrogen, wherein n is zero or an
Figure imgf000006_0004
integer of 1 or 2 and R4 is hydrogen or hydroxyl,
CH3-,
H2N-(CH2)4-,
HO2C-(CH2)m- wherein m is an integer of 1 or 2, or
Figure imgf000006_0005
2 wherein m is as defined above,
HO2CCH (OH) CH
Figure imgf000006_0006
HO2C-CH2
Figure imgf000006_0007
Figure imgf000007_0003
X is O, S, or NH, and
R2 is alkyl of from one to six carbon atoms ; - wherein R5 is
Figure imgf000007_0001
R5
Figure imgf000007_0002
wherein R6, R7, R8, or R9 are each independently hydrogen, alkyl of from one to six carbon atoms, alkoxy of from one to six carbon atoms, halogen, or trifluoromethyl. wherein R1 and X are as defined above,
wherein R1 and X are as defined above,
wherein R1 is as defined above,
wherein X and R1 are as defined above,
wherein R1 and X are as defined above, or
Figure imgf000008_0001
wherein R1 and X are as
Figure imgf000009_0001
defined above;
G is wherein R5 is as defined above, or
wherein R10 is
Figure imgf000009_0002
hydrogen.
alkyl of from one to six carbon atoms ,
- CO2CH3 ,
Figure imgf000009_0003
-CH2-CH=CH2,
-CH2-C≡CH,
-CH2-CN,
-CH2-OH,
Figure imgf000009_0004
-CH2-CH2X-R1 wherein X and R1 are as defined above,
-CH2X-R1 wherein X and R1 are as defined above,
wherein X and R1 are as defined
Figure imgf000009_0005
above,
- CH2-CH2CH2CHO-N H2 ,
-CH2-CH2-S (O) n-R1 wherein n and R1 are as defined above,
-(CH2)n-CONH2 wherein n is as defined above,
Figure imgf000010_0001
wherein R1 is as defined above,
Figure imgf000010_0002
wherein X and R1 are as defined above;
alternatively, E-G is
Figure imgf000010_0003
wherein R1, X, R5, and R10 are as defined above;
wherein R11 is
Figure imgf000010_0004
hydrogen,
alkyl,
Figure imgf000011_0001
R12 is
•
Figure imgf000011_0002
wherein R1 and X are as defined above, wherein R1 and X are as defined
Figure imgf000011_0003
above,
Figure imgf000011_0004
-CH2-OC2H5 and R1 and X are as defined above and p is zero or an integer of one, or wherein R14 is
Figure imgf000011_0005
or
Figure imgf000011_0006
-OC2H5
and R11 and p are as defined above;
provided R1 with the exclusion of R1 being hydrogen is encompassed within the definition of at least one of A, E, G, or J; or a pharmaceutically acceptable salt thereof. As amino acid prodrug renin inhibitors the compounds of Formula I are useful for the treatment of hypertension, hyperaldosteronism, congestive heart failure, and diseases caused by retroviruses
including HTLC I, II, and III as well as diagnostic tools for the identification of cases of hypertension due to renin excess.
A still further embodiment of the present invention is a pharmaceutical composition for
administering an effective amount of a compound of Formula I in unit dosage form in the treatment methods mentioned above.
Finally, the present invention is directed to methods for production of a compound of Formula I.
DETAILED DESCRIPTION OF THE INVENTION
In the compounds of Formula I, the term "alkyl" means a straight or branched hydrocarbon radical having from one to six carbon atoms and includes, for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, n-hexyl, and the like.
"Alkoxy" is O-alkyl as defined above for alkyl. "Halogen" is fluorine, chlorine, bromine, or iodine.
The following table provides a list of
abbreviations and definitions thereof used in the present inventions.
Abbreviation Amino Acid
Ala Alanine
Asp Aspartic acid * If the optical activity of the amino acid is
other than L(S), the amino acid or abbreviation is preceded by the appropriate configuration D(R) or DL(RS). Abbreviation Amino Acid
Glu Glutamic acid
Gln Glutamine
Gly Glycine
Lys Lysine
Phe Phenylalanine
Pro Proline
Ser Serine
Thr Threonine
Tyr Tyrosine
Abbreviation Modified and Unusual Amino
Acid
Alg 2-Amino-4-pentenoic acid
(Allylglycine)
Atm 2-Amino-3-(2-amino-5- thiazole)propanoic acid
Chx Cyclohexylalanine
(Hexahydrophenylalanine)
Hse 2-Amino-4-hydroxybutyric acid (Homoserine)
Mal 2-Amino-1,3-propanedioic acid, monomethyl ester
Pgy 2-Aminopentanoic acid
(Propylglycine)
Amides With
2-(2-Aminoethyl)pyridine
N-(2-Aminoethyl)morpholine
2-Aminomethylpyridine
2-Methylbutylamine
Figure imgf000013_0001
-NHCH2CH2N(CH2CH2OH)2 2-Bis(2-hydroxyethyl)
amino)etholamine Abbreviation Esters With
Morpholine
Figure imgf000014_0001
-OCH3 Methanol
-OC2H5 Ethanol
-OCH(CH3)2 2-Propanol
-OC(CH3)3 tertiary Butanol
-OBz Benzyl alcohol
Protecting Group
Cbz Benzyloxycarbonyl
Boc tertiary Butyloxycarbonyl TBDMS tertiary Butyldimethylsilyl
Miscellaneous
CAD 2 (S)-Amino-1-cyclohexyl-6- methyl-3(R),4(S)- heptanediol
SMO N-Morpholinesulphonyl
Ph Phenyl
Bz Benzyl
Abbreviation Solvents and Reagents
DMF N,N-Dimethylformamide
DMAP 4-Dimethylamino pyridine
DMSO Dimethylsulfoxide
HOBT Hydroxybenzotriazole
DCC N,N'-Dicyclohexyl- carbodiimide
DCU 1,3-Dicyclohexylurea
HOAc Acetic acid
Et3N Triethylamine
THF Tetrahydrofuran
CH2Cl2 Dichloromethane
CHCl3 Chloroform
Et2O Diethyl ether
CDI Carbonyl Diimidazole
TBAF Tetra-n-butylammonium
fluoride
KBH4 Potassium Borohydride
DMAP N,N-Dimethylaminopyridine EtOAc Ethyl Acetate
NaCl Sodium Chloride Abbreviation Solvents and Reagents
Na2SO3 Sodium Sulfite
NaHCO3 Sodium Bicarbonate
NaIO4 Sodium periodate
Na2CO3 Sodium Carbonate
MgSO4 Magnesium Sulfate
HCl Hydrochloric acid
EtOH Ethanol
MeOH Methanol
D2O Deuterium Oxide
Na2SO4 Sodium Sulfate
TFA Trifluoroacetic acid
TsOH Para-toluenesulfonic acid An amino acid or organic acid within the bracket in a structure means the bracketed moiety is attached via the α carboxylic acid or carboxylic acid,
respectively, to the free hydroxyl or phenolic group of the preceding amino acid or organic alcohol (where there is more than one free hydroxyl or phenolic group in the preceding moiety the amino acid or organic acid may be attached to either or both) to form an ester such as, for example:
Ser(Phe),
Ser(Gln),
Ser(Pro),
Ser(Glu),
Ser(Lys),
Ser(Asp),
Ser(Gly),
Ser(Ala),
Ser(COCH2CH2CO2H),
Ser(P(O) (OH)2),
Ser (COCH(OH)CH(OH)CO2H),
Thr(Phe),
Thr(Gln),
Thr(Pro),
Thr(Glu),
Thr(Lys), Thr(Asp),
Thr(Gly),
Thr(Ala),
Thr(COCH2CH2CO2H),
Thr(P(O) (OH)2),
Thr(COCH(OH)CH(OH)CO2H),
Tyr(Phe),
Tyr(Gln),
Tyr(Pro),
Tyr(Glu),
Tyr(Lys),
Tyr(Asp),
Tyr(Gly),
Tyr(Ala),
Tyr(COCH2CH2CO2H),
Tyr(P(O) (OH)2),
Tyr(COCH (OH) CH (OH) CO2H), , Hse(Phe),
Hse(Gln),
Hse(Pro),
Hse(Glu),
Hse(Lys),
Hse(Asp),
Hse(Gly),
Hse(Ala),
Hse(COCH2CH2CO2H),
Hse(P(O) (OH)2),
Hse (COCH (OH) CH (OH) CO2H), CAD(Phe),
CAD(Gln),
CAD(Pro),
CAD(Glu),
CAD(Lys),
CAD(Asp),
CAD(Gly),
CAD(Ala),
CAD(COCH2CH2CO2H), CAD(P(O)(OH)2),
CAD(COCH(OH)CH(OH)CO2H),
CAD(2•Phe),
CAD(2•Gln),
CAD(2•Pro),
CAD(2•Glu),
CAD(2•Lys),
CAD(2•Asp),
CAD(2•Gly),
CAD(2•Ala),
CAD(2•COCH2CH2CO2H),
CAD(2•P(O)(OH)2),
CAD(2•COCH(OH)CH(OH)CO2H),
(Phe)
(Gln)
(Pro)
(Glu)
(Lys)
(Asp)
(Gly)
(Ala)
Figure imgf000017_0001
(HO2CCH2CH2CO)O
((HO)2P(O))OCH
Figure imgf000017_0002
(HO2CCH(OH)CH(OH)CO)O- -
Figure imgf000018_0005
0
(Phe)O-CH2-C(CH3)
(Glu)O-CH2-C(CH3)
(Pro)O-CH2-C(CH3) (Glu)O-CH2-C(CH3)
(Lys)O-CH2-C(CH3)
(Asp)O-CH2-C(CH3)2 O
(Gly)O-CH2-C(CH3)2-
(Ala)O-CH2-C(CH3)2-
Figure imgf000018_0006
(HO2CCH2CH2CO)O-CH2-C(CH3)2
Figure imgf000018_0004
-
((HO)2P(O))O-CH2-C(CH3)
Figure imgf000018_0003
(HO2CCHCOH)CH(OH)CO)O-CH2-C(CH3) -
Figure imgf000018_0002
Figure imgf000018_0001
Figure imgf000019_0001
However, when the preceding unbracketed moiety is an organic amine, the bracketed amino acid or organic acid forms an amide bond with the amine such as, for example:
Atm(Phe),
Atm(Gln),
Atm(Pro),
Atm(Glu),
Atm(Lys),
Atm(Asp),
Atm(Gly),
Atm(Ala),
Atm(COCH2CH2CO2H),
Atm(P(O)(OH)2),
Atm(COCH(OH)CH(OH)CO2H),
For purposes of the present invention a
"prodrug" refers to a compound of Formula I which is biotransformed into the active renin inhibitor in a mammal.
The compounds of Formula I are capable of further forming pharmaceutically acceptable acid addition and/or base salts. All of these forms are within the scope of the present invention.
Pharmaceutically acceptable acid addition salts of the compound of Formula I include salts derived from nontoxic inorganic acids such as hydrochloric, nitric, phosphoric, sulfuric, hydrobromic, hydriodic, phosphorous and the like, as well as the salts derived from nontoxic organic acids such as aliphatic mono- and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxy alkanoic acids, alkanedioic acids, aromatic acids, aliphatic and aromatic
sulfonic acids, etc. Such salts thus include
sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogenphosphate,
dihydrogenphosphate, metaphosphate, pyrophosphate. chloride, bromide, iodide, acetate, propionate, caprylate, isobutyrate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, mandelate, benzoate, chlorobenzoate, methylbenzoate,
dinitrobenzoate, phthalate, benzenesulfonate,
toluenesulfonate, phenylacetate, citrate, lactate, maleate, tartrate, methanesulfonate and the like.
Also contemplated are salts of amino acids such as arginate and the like and gluconate, galacturonate (see, for example. Bergs S. M., et al,
"Pharmaceutical Salts," Journal of Pharmaceutical Science, Vol. 66, pages 1-19 (1977)).
The acid addition salts of said basic compounds are prepared by contacting the free base form with a sufficient amount of the desired acid to produce the salt in the conventional manner. The free base form may be regenerated by contacting the salt form with a base and isolating the free base in the conventional manner. The free base forms differ from their respective salt forms somewhat in certain physical properties such as solubility in polar solvents, but otherwise the salts are equivalent to their
respective free bases for purposes of the present invention.
Pharmaceutically acceptable base addition salts of acidic compounds are formed with metals or amines such as alkali and alkaline earth metals or organic amines. Examples of metals used as cations are sodium, potassium, magnesium, calcium and the like. Examples of suitable amines are
N,N'-dibenzylethylenediamine, chloroprocaine,
choline, diethanolamine, ethylenediamine,
N-methylglucamine, and procaine (see, for example, Berge S. M., et al. Journal of Pharmaceutical
Science, Vol. 66, pages 1-19 (1977)).
The base addition salts of said acidic compounds are prepared by contacting the free acid form with a sufficient amount of the desired base to produce the salt in the conventional manner. The free acid form may be regenerated by contacting the salt form with an acid and isolating the free acid in the
conventional manner. The free acid forms differ from their respective salt forms somewhat in certain physical properties such as solubility in polar solvents, but otherwise the salts are equivalent to their respective free acids for purposes of the present invention.
Certain of the compounds of the present
invention can exist in unsolvated forms as well as solvated forms, including hydrated forms. In
general, the solvated forms, including hydrated forms, are equivalent to unsolvated forms and are intended to be encompassed within the scope of the present invention.
The compounds of the present invention possess one or more chiral centers and each center may exist in the R(D) or S(L) configuration. The present invention includes all enantiomeric forms as well as the appropriate mixtures thereof.
A preferred compound of Formula I is one wherein A is
wherein R is hydrogen or alkyl of from one to six carbon atoms,
Figure imgf000022_0001
Figure imgf000023_0001
3 - wherein R1 is
Figure imgf000023_0002
- wherein R3 is
Figure imgf000023_0003
hydrogen,
Figure imgf000023_0004
CH3-,
H2N-(CH2)4-,
HO2C-(CH2)m- wherein m is an integer of
1 of 2, or
Figure imgf000024_0004
2 wherein m is as defined above,
Figure imgf000024_0001
R2 is alkyl of from one to six carbon atoms;
E is wherein R5 is
Figure imgf000024_0002
wherein R6, R7, R8, or R9 are each independently hydrogen,
Figure imgf000024_0003
alkyl of from one to six carbon atoms, alkoxyl of from one to six carbon atoms, halogen or trifluoromethyl,
Figure imgf000025_0001
wherein R1 is
hydrogen, wherein R3 is
Figure imgf000025_0002
hydrogen,
Figure imgf000025_0003
CH3-,
H2N-(CH2)4-,
HO2C-(CH2)m- wherein m is an integer of 1 or 2, or
Figure imgf000025_0004
wherein m is as defined above,
HO 2C-CH(OH)CH(OH)
Figure imgf000025_0005
Figure imgf000026_0001
G is C- wherein R5 is
Figure imgf000026_0002
Figure imgf000026_0003
wherein R6, R7, R8, or R9 are each independently hydrogen, alkyl of from one to six carbon atoms, alkoxy of from one to six carbon atoms, halogen, or trifluoromethyl.
Figure imgf000026_0004
Figure imgf000027_0001
wherein X is O, S, or NH and R1 is as defined above
Figure imgf000027_0002
wherein R1 and X are as defined above.
wherein R1 is as defined above.
Figure imgf000027_0003
wherein X and R1 are as defined above, wherein R1 and X are as defined above,
wherein R1 and X are as
Figure imgf000028_0001
defined above, or wherein R10 is
Figure imgf000028_0002
hydrogen,
alkyl of from one to six carbon atoms , -CO2CH3,
Figure imgf000028_0003
-CH2-CH=CH2,
-CH2-C≡CH,
-CH2-CN,
-CH2-OH,
- 3 ,
Figure imgf000028_0004
-CH2-CH2X-R1 wherein X and R1 are as defined above,
-CH2X-R1 wherein X and R1 are as defined above,
-X- wherein X and R1 are as defined
Figure imgf000028_0005
above,
-CH2-CH2CH2CH2-NH2,
-CH2-CH2-S(O)n-R1 wherein n is zero or an integer of 1 or 2 and R1 is as defined above, -(CH2)n-CONH2 wherein n is as defined above,
Figure imgf000029_0001
wherein R1 is as defined above,
Figure imgf000029_0002
wherein X and R are as defined above;
alternatively, E-G is
Figure imgf000029_0003
wherein R1, X, R5, and R10 are as defined above;
J is wherein R11 is
Figure imgf000029_0004
hydrogen,
alkyl,
Figure imgf000030_0001
R12 is wherein R1 is as defined above.
Figure imgf000030_0002
wherein R1 is as defined
R1 above,
Figure imgf000030_0003
Figure imgf000030_0004
-CH2-OC2H5 and R1 and X are as defined above or 4 wherein R14 is
Figure imgf000030_0005
Figure imgf000030_0006
-OC2H5 and R11 is as defined above; provided
R1 with the exclusion of R1 being hydrogen is encompassed within the definition of at least one of A, E, G, or J.
A more preferred compound of Formula I is one wherein J is
wherein R11 is
Figure imgf000030_0007
alkyl,
Figure imgf000031_0001
R12 is 2
and
Figure imgf000031_0002
R1 is hydrogen, - wherein R3 is
Figure imgf000031_0003
hydrogen,
Figure imgf000031_0004
H2N-(CH2)4-,
HO2C-(CH2)m- wherein m is an integer of
1 or 2, or
Figure imgf000031_0005
wherein m is as defined above,
Figure imgf000032_0001
and X is O,
Figure imgf000032_0002
S, or NH, or wherein R14 is
, or
Figure imgf000032_0003
-OC2H5 and
R11 is as defined above.
Particularly valuable are:
SMO-Phe-Ser(Phe)-CAD;
SMO-Phe-Ser(Gln)-CAD;
SMO-Phe-Ser(Pro)-CAD; SMO-Phe-Ser(Glu)-CAD;
SMO-Phe-Ser(Lys)-CAD;
SMO-Phe-Ser(Asp)-CAD;
SMO-Phe-Ser(Gly)-CAD;
SMO-Phe-Ser(Ala)-CAD;
SMO-Phe-Ser(COCH2CH2CO2H)-CAD;
SMO-Phe-Ser(P(O)(OH)2)-CAD;
SMO-Phe-Ser(COCH(OH)CH(OH)CO2H)-CAD; SMO-Phe-Thr(Phe)-CAD;
SMO-Phe-Thr(Gln)-CAD;
SMO-Phe-Thr(Pro)-CAD;
SMO-Phe-Thr(Glu)-CAD;
SMO-Phe-Thr(Lys)-CAD;
SMO-Phe-Thr(Asp)-CAD;
SMO-Phe-Thr(Gly)-CAD;
SMO-Phe-Thr(Ala)-CAD;
SMO-Phe-Thr(COCH2CH2CO2H)-CAD;
SMO-Phe-Thr(P(O)(OH)2)-CAD;
SMO-Phe-Thr(COCH(OH)CH(OH)CO2H)-CAD; Boc-Tyr(Phe)-Pgy-CAD;
Boc-Tyr(Gln)-Pgy-CAD;
Boc-Tyr(Pro)-Pgy-CAD;
Boc-Tyr(Glu)-Pgy-CAD;
Boc-Tyr(Lys)-Pgy-CAD;
Boc-Tyr(Asp)-Pgy-CAD;
Boc-Tyr(Gly)-Pgy-CAD;
Boc-Tyr(Ala)-Pgy-CAD;
Boc-Tyr(COCH2CH2CO2H)-Pgy-CAD;
Boc-Tyr(P(O)(OH)CH(OH)CO2H)-Pgy-CAD; Boc-Tyr(COCH(OH)CH(OH)CO2H)-Pgy-CAD; SMO-Phe-Hse(Phe)-CAD;
SMO-Phe-Hse(Gln)-CAD
SMO-Phe-Hse(Pro)-CAD
SMO-Phe-Hse(Glu)-CAD
SMO-Phe-Hse(Lys)-CAD
SMO-Phe-Hse(Asp)-CAD
SMO-Phe-Hse(Gly)-CAD SMO-Phe-Hse(Ala)-CAD;
SMO-Phe-Hse(COCH2CH2CO2H)-CAD;
SMO-Phe-Hse(P(0) (OH)2)-CAD;
SMO-Phe-Hse(COCH(OH)CH(OH)CO2H)-CAD;
SMO-Phe-Mal-CAD(2•Phe);
SMO-Phe-Mal-CAD(2•Gln);
SMO-Phe-Mal-CAD(2•Pro);
SMO-Phe-Mal-CAD(2•Glu);
SMO-Phe-Mal-CAD(2•Lys);
SMO-Phe-Mal-CAD(2•Asp);
SMO-Phe-Mal-CAD(2•Gly);
SMO-Phe-Mal-CAD(2•Ala);
SMO-Phe-Mal-CAD(2•COCH(OH)CH(OH)CO2H);
SMO-Phe-Mal-CAD(2•P(O) (OH)2);
SMO-Phe-Mal-CAD(2•COCH(OH)CH(OH)CO2H);
SMO-Phe-Atm-CAD(2•Phe);
SMO-Phe-Atm-CAD(2•Gln);
SMO-Phe-Atm-CAD(2•Pro);
SMO-Phe-Atm-CAD(2•Glu);
SMO-Phe-Atm-CAD(2•Lys);
SMO-Phe-Atm-CAD(2•Asp);
SMO-Phe-Atm-CAD(2•Gly);
SMO-Phe-Atm-CAD(2•Ala);
SMO-Phe-Atm-CAD(2•COCH2CH2CO2H);
SMO-Phe-Atm-CAD(2•P(O) (OH)2);
SMO-Phe-Atm-CAD(2•COCH(OH)CH(OH)CO2H);
SMO-Phe-Atm(Phe)-CAD;
SMO-Phe-Atm(Gln)-CAD;
SMO-Phe-Atm(Pro)-CAD;
SMO-Phe-Atm(Glu)-CAD;
SMO-Phe-Atm(Lys)-CAD;
SMO-Phe-Atm(Asp)-CAD;
SMO-Phe-Atm(Gly)-CAD;
SMO-Phe-Atm(Ala)-CAD;
SMO-Phe-Atm(COCH2CH2CO2H)-CAD;
SMO-Phe-Atm(P(O) (OH)2)-CAD;
SMO-Phe-Atm(COCH(C)H)CH(OH)CO2H)-CAD; (Phe)O- -Alg-CAD;
(Gln)O- -Alg-CAD;
(Pro)O- -Alg-CAD;
(Glu)O- -Alg-CAD;
(Lys)O- -Alg-CAD;
(Asp)O- -Alg-CAD;
(Gly)O- -Alg-CAD;
(Ala)O- -Alg-CAD;
Figure imgf000035_0001
(HO2CCH2CH2CO)O-CH2-
Figure imgf000035_0002
-Phe-Alg-CAD;
((HO)2P(O))OCH2- -Phe-Alg-CAD;
Figure imgf000035_0003
(HO2CCH(OH)CH(OH)CO)OCH2
Figure imgf000035_0004
-Phe-Alg-CAD;
(Phe)O-CH2-C(CH3) -Phe-Alg-CAD;
(Gln)O-CH2-C(CH3) - -Phe-Alg-CAD;
(Pro)O-CH2-C(CH3) -Phe-Alg-CAD;
(Glu)O-CH2-C(CH3) -Phe-Alg-CAD;
Figure imgf000035_0005
(Lys)O-CH2-C(CH3) -Phe-Alg-CAD;
(Asp)O-CH2-C(CH3) -Phe-Alg-CAD;
(Gly)O-CH2-C(CH3) -Phe-Alg-CAD;
(Ala)O-CH2-C(CH3) - -Phe-Alg-CAD;
Figure imgf000036_0001
(HO2CCH2CH2CO)O-CH2-C(CH3)2
Figure imgf000036_0002
-Phe-Alg-CAD;
((HO)2P(O))O-CH2-C(CH3)2 Phe-Alg-CAD;
Figure imgf000036_0003
(HO2CCH(OH)CH(OH)CO)O-CH2-C(CH3)2- -Phe-Alg-CAD;
Figure imgf000036_0004
(Phe)O-CH2 Phe-Atm-CAD;
(Gln)O-CH2 Phe-Atm-CAD;
(Pro)O-CH2 Phe-Atm-CAD;
(Glu)O-CH2 Phe-Atm-CAD;
(Lys)O-CH2 Phe-Atm-CAD;
(Asp)O-CH2 Phe-Atm-CAD;
(Gly)O-CH2 Phe-Atm-CAD;
(Ala)O-CH2
Figure imgf000036_0005
Phe-Atm-CAD;
(HO2CCH2CH2CO)O-CH2-
Figure imgf000036_0006
-Phe-Atm-CAD; ((HO)2P(O)O-CH2 -Phe-Atm-CAD;
Figure imgf000037_0001
(HO2CCH(OH)CH(OH)CO)O-CH2- -Phe-Atm-CAD;
Figure imgf000037_0002
(Phe)O-CH2-C(CH3) -Phe-Atm-CAD;
(Gln)O-CH2-C(CH3) -Phe-Atm-CAD;
(Pro)O-CH2-C(CH3) -Phe-Atm-CAD; (Glu)O-CH2-C(CH3) -Phe-Atm-CAD;
(Lys)O-CH2-C(CH2)2 -Phe-Atm-CAD;
(Asp)O-CH2-C(CH3) -Phe-Atm-CAD;
(Gly)O-CH2-C(CH3) -Phe-Atm-CAD;
(Ala)O-CH2-C(CH3) -Phe-Atm-CAD;
Figure imgf000037_0003
(HO2CCH2CH2CO)O-CH2-C(CH3)2
Figure imgf000037_0004
-Phe-Atm-CAD;
((HO)2P(O))O-CH2-C(CH3)2 -Phe-Atm-CAD;
Figure imgf000037_0005
(HO2CCH(OH)CH(OH)CO)O-CH2-C(CH3)2
Figure imgf000037_0006
-Phe-Atm-CAD;
Figure imgf000037_0007
Figure imgf000038_0001
or a pharmaceutically acceptable salt thereof.
Amidon, G.L., et al, Journal of Pharmaceutical Sciences, Vol. 69, pages 1363-1368 (1980) described a procedure for improving the intestinal absorption of water - insoluble drugs. This strategy converts the insoluble compound to a soluble derivative which subsequently is converted enzymatically to the parent compound in vivo. Thus, the soluble derivatives of the water insoluble drug acts as a prodrug which is converted by enzymes in the surface coat of the brush border region of the microvillous membrane in the intestine to the desired drug. The ability of a compound of Formula I which is a water soluble amino acid derivative of a renin inhibitor to act as a prodrug that is converted by intestinal enzymes to the parent renin inhibitor is tested using the following procedure.
Stability in Intestinal Perfusate - Perfusate is generated by oscillating 15 to 20 mL MES buffer through rat jejunum in situ at 30 mL/min using a withdrawal/infusion pump (Harvard Apparatus,
Model 4200-015). At 1.5 h, the perfusate is
collected and spiked with prodrug at 37 μg/mL. The solution is incubated at 37°C and disappearance of prodrug over time is monitored by direct injection (25 μL) onto HPLC.
Stability in Suspensions of Brush Border
Membranes (BBM) - BBM are prepared from rat or rabbit small intestine by the method of Kessler, et al,
Biochim. Biophys. Acta 506:136-154, (1978) with some modifications. Vesicles (50 μL) are combined with 450 μL drug or prodrug (10 μg/mL) and incubated at 37°C. At selected time points, 100 μL prodrug/BBM suspension is removed and diluted with 100 μL
acetonitrile, then 150 μL MES, centrifuged, and 50 μL is injected onto HPLC. HPLC Conditions - Samples are analyzed by high pressure liquid chromatography using a two pump binary-gradient system (HP 1090 Liquid
Chromatograph), a Lambda-Max LC Spectrophotometer (Waters, Model 481) set to 214 nm, and a ChromJet integrator (SpectraPhysics). The Econosil column (C8, 4.6 x 250 mm) is at room temperature. Solvent A is water/acetonitrile/triethylamine (TEA) (90:10:0.1) and solvent B is acetonitrile/water/TEA (70:30:0.1). The mobile phase is degassed and maintained with zero grade helium. Isocratic conditions consisted of 65% solvent A and 35% solvent B. At a flow rate of
1.0 mL/min, the retention times are 56 min for the drug and 9.6 min for the prodrug. MES buffer is 10 mM (2-[N-Morpholino]ethanesulfonic acid, made iso-osmotic with 10 mM KCL and 140 mM NaCl and adjusted to pH 6.5 with IN NaOH.
The data in Table I show some representative water soluble amino acid prodrug renin inhibitors which are converted to the parent renin inhibitor by intestinal enzymes and also by brush border enzymes.
Figure imgf000041_0001
A method of preparing a compound having the Formula I
A-E-G-J
wherein A is
wherein R is hydrogen or alkyl of from one to six carbon atoms. -N- - wherein R is as defined above,
Figure imgf000042_0001
Figure imgf000042_0002
wherein R is as defined above.
Figure imgf000042_0003
Figure imgf000043_0001
3
Figure imgf000043_0002
wherein R1 is
hydrogen, wherein R3 is
Figure imgf000043_0003
hydrogen,
wherein n is zero or an integer of 1 or 2 and R4 is hydrogen or
Figure imgf000043_0004
hydroxyl,
CH3-,
H2N-(CH2)4-,
HO2C-(CH2)m- wherein m is an integer of 1 or 2, or wherein m is as defined
Figure imgf000043_0005
above,
HO2CCH(OH)CH(OH
Figure imgf000043_0006
Figure imgf000044_0001
X is O, S, or NH, and
R2 is alkyl of from one to six carbon atoms;
E is wherein R5 is
Figure imgf000044_0002
wherein R6, R7, R8, or R9 are each independently hydrogen, alkyl of from one to six carbon atoms, alkoxy of from one to six carbon atoms, halogen, or
trifluoromethyl,
wherein R1 and X are as defined above,
wherein R1 and X are as defined above,
wherein R1 is as defined above,
wherein X and R1 are as defined above,
Figure imgf000045_0001
2 wherein R1 and X are as defined above, or wherein R1 and X are as defined above;
Figure imgf000046_0001
G is N wherein R5 is as defined above, or
wherein R10 is
Figure imgf000046_0002
hydrogen,
alkyl of from one to six carbon atoms,
-CO2CH3,
Figure imgf000046_0003
-CH2-CN,
-CH2-OH,
- - 3
Figure imgf000046_0005
Figure imgf000046_0004
-CH2-CH2X-R1 wherein X and R1 are as defined above,
-CH2X-R1 wherein X and R1 are as defined above. wherein X and R1 are as defined
Figure imgf000047_0004
above,
-CH2-CH2CH2CH2-NH2,
-CH2-CH2-S(O)n-R1 wherein n and R1 are as defined above,
-(CH2)n-CONH2 wherein n is as defined above,
Figure imgf000047_0001
wherein R1 is as defined above,
Figure imgf000047_0002
wherein X and R1 are as defined above;
alternatively, E-G is
Figure imgf000047_0003
wherein R1, X, R5, and R10 are as defined above; J is wherein R11 is
Figure imgf000048_0001
hydrogen,
alkyl,
R12 is
Figure imgf000048_0002
3
wherein R1 and X are as defined above, wherein R1 and X are as defined above,
Figure imgf000048_0003
Figure imgf000048_0004
-CH2-OC2H5 and R1 and X are as defined above and p is zero or an integer of one, or wherein R14 is
Figure imgf000048_0005
or
Figure imgf000049_0001
-OC2H5
and R11 and p are as defined above;
provided R1 with the exclusion of R1 being hydrogen is encompassed within the definition of at least one of A, E, G, or J; or a pharmaceutically acceptable salt thereof comprises:
a) coupling a compound of Formula II
A'-E'-G'-J'
II wherein A', E', G', and J' are as defined above for A, E, G, and J provided R1 is hydrogen and is encompassed within the definition of at least one of A', E', G', or J' with a compound of Formula III
R1a-XH
III wherein R1a is as defined above for R1 but excluding R1 is hydrogen and providing any basic or acidic groups contain conventional protecting groups and X is as defined above to afford a compound of Formula IV
A''-E''-G''-J'
IV wherein A'', E'', G'', and J'' are as defined above for A, E, G, and J provided R1a is as defined above and is encompassed within the definition of at least one of A", E'' , G" or J";
b) a compound of Formula IV is deprόtected in a conventional manner to afford a compound of
Formula I; and if desired, converting a compound of Formula I to a corresponding pharmaceutically
acceptable salt by conventional means and, if so desired, converting the corresponding
pharmaceutically acceptable salt to a compound of Formula I by conventional means.
A compound of Formula II may be prepared by sequential stepwise or fragment coupling of the amino acids or fragments selected from A', E', G', or J' to the preceding amino acid or fragment using
conventional peptide synthesis methodology such as, for example, solution peptide synthesis or solid phase peptide synthesis to afford a compound of
Formula II.
A compound of Formula IV is prepared from anino acids or fragments selected from A'', E'', G" , or J'' using the methodology used to prepare a compound of Formula II.
A compound of Formula III is either known or capable of being prepared by methods known in the art.
Coupling methods that can be employed in
preparing the compounds of Formula I are discussed in "The Peptides. Analysis, Synthesis, Biology,"
Gross, E., and Meienhofer, J., eds. Academic Press, New York, New York, Vol. 1, 1979. Further,
protecting groups that may be employed in the
preparation of a compound of Formula I, as well as methods for incorporation and removal of these protecting groups are discussed in "The Peptides. Analysis, Synthesis, Biology," Gross, E., and Meienhofer, J., eds., Academic Press, New York, New York, Vol. 3, 1981.
Figure imgf000052_0001
Figure imgf000053_0001
Figure imgf000054_0001
Figure imgf000055_0001
Figure imgf000056_0001
Figure imgf000057_0001
Figure imgf000058_0001
Figure imgf000059_0001
Figure imgf000060_0001
Preferred methods for the preparation of
compounds of Formula I are described in Schemes 1 to 7.
Compounds of Formula I are designated by numbers 8, 11, 14, 24a, 24b, 30a, 30b, 40, 43, 54, an 58.
These schemes illustrate preferred methods from which a person, skilled in the art of organic
chemistry, could analogously prepare all compounds of Formula I.
Compounds of Formula I designated 8, 11, and 14 are prepared from a compound of Formula 1 as outlined in Scheme I. Thus, SMO-Cl(l) (morpholinosulfamyl chloride) (prepared according to the method of R.
Wegler and K. Bodenbenner, Annalen der Chemie, 624, 25 (1959) ) is reacted with Phe in the presence of a base such as, for example, Et3N and the like and a solvent such as, for example, CH2Cl2 and the like at about -30°C to about 100°C for about one to 24 hours to give the compound of Formula 2. Preferably, the reaction is carried out in the presence of Et3N and CH2Cl2 at about 25°C for about 2 to 6 hours.
Treatment of Cbz-Ser (3) with CAD in the presence of a coupling reagent such as, for example, DCC and HOBT and the like and a solvent such as, for example, CH2Cl2, CHCl3, THF and the like optionally in the presence of an aprotic solvent such as, for example, DMF, DMSO and the like at about 0°C to about 25°C for about 6 hours to about 5 days to give the compound of Formula 4. The compound of Formula 4 is treated with hydrogen in the presence of a catalyst such as, for example, 5% to 20% palladium on carbon, platinum oxide, 5% to 20% platinum on carbon, and the like in a solvent such as, for example, EtOH, EtOAc, THF, dioxane, DMF and the like at about 0°C to about 100°C for about 30 minutes to about 24 hours to give the compound of Formula 5. The compound of Formula 2 is reacted with the compound of Formula 5 in the presence of a coupling reagent such as, for example, DCC and HOBT and the like and a solvent such as, for example, CHCl3, CH2Cl2, THF, dioxane, EtOAc, DMF, DMSO and the like at about 0°C to about 40°C for about 1 to 5 days to give the compound of Formula 6. The compound of Formula 6 is reacted with LYS.2BOC in the presence of a coupling reagent such as, for example, CDI, DCC, and the like in the presence of a solvent such as, for example, CHCl3, CH2Cl2, THF, dioxane, EtOAc, DMF, DMSO and the like at about 0°C to about 100°C for about 3 hours to about 3 days to give the compound of Formula 7. The compound of Formula 7 is treated with an acid such as, for example, trifluoroacetic acid, or a nonpolar inert solvent such as CH2Cl2 saturated with hydrogen chloride gas at about -20°C to about 25°C for about
1 to 5 hours to give the compound of Formula 8.
In a similar manner the compound of Formula 6 is reacted with Cbz-Gln or to afford,
Figure imgf000062_0001
respectively, a compound of Formula 10 and a compound of Formula 13. Compounds of Formula 10 and 13 are reacted with hydrogen in the presence of a catalyst such as for example 50% palladium on carbon and an acid such as for example TFA and the like in a solvent such as for example ethanol and the like toafford, respectively, a compound of Formula 11 and a compound of Formula 14.
Compounds of Formula I designated 24a and 24b are prepared from a compound of Formula 2 as outlined in Scheme 2. Thus, the compound of Formula 2 is reacted with a coupling reagent such as, for example, CDI and the like in a solvent such as THF, CH2Cl2, mixtures thereof and the like followed by
O,N-dimethylhydroxylamine hydrochloride and a base such as, for example, N-methylpiperidine and the like to give the compound of Formula 15. The compound of Formula 15 is added to the magnesium Grignard reagent of 4-bromo-1-butene in a solvent such as, for
example, THF, Et2O and the like to give the compound of Formula 16. The compound of Formula 16 is treated with a hydride reagent such as, for example, KBH4 and the like in a solvent such as, for example, absolute EtOH and the like to give the compound of Formula 17 as a mixture of R and S isomers at the carbon atom towhich the hydroxyl group is attached. Choice of solvent and reducing reagent can vary the ratio of isomers obtained to give a predominant excess of a particular desired isomer. The R and S isomers of the compound of Formula 17 may be separated into the individual R and S isomers by conventional separation techniques such as, for example, chromatography, fractional crystallization and the like.
Alternatively, the compound of Formula 17 (R and S mixture of isomers) is reacted with
t-butyldimethylsilyl chloride (TBDMS-Cl) in the presence of imidazole and DMF to give the compound of Formula 18 as a mixture of R and S isomers at the carbon atom to which the O-TBDMS group is attached. The R and S isomers of the compound of Formula 18 may be separated into the individual R and S isomers by conventional separation techniques such as, for example, chromatography and the like. Additionally, other hydroxyl protecting groups such as, for
example, acetyl, benzyl, and the like may be used in place of the TBDMS group with subsequent separation of the R and S protected hydroxyl groups following the methodology used to separate the isomers of the compound of Formula 18. Either the compound of Formula 18a (R-isomer) or Formula 18b (S-isomer) is reacted with a fluoride ion source such as, for example, t-butylammonium fluoride and the like in a solvent such as, for example, THF and the like to give the compound of Formula 19a (R-isomer) or Formula 19b (S-isomer). Either the compound of Formula 19a (R-isomer) or Formula 19b (S-isomer) is reacted with the imidazolide of N-α-Boc-ε-CBZ-LYS (prepared from NCC-Boc-ε-Cbz-Lys, CDI and DMAP)
(optionally compatible N-protecting groups other than Boc or Cbz may also be used to protect the amino groups of Lys) in a solvent such as, for example, CH2Cl2 and the like at about ambient temperature to give the compound of Formula 20a (R-isomer) or
Formula 20b (S-isomer). Either the compound of
Formula 20a (R-isomer) or Formula 20b (S-isomer) is treated with an oxidizing reagent such as, for example, NaIO4 with a catalytic amount of RuO2, and the like to give the compound of Formula 21a (R- isomer) or Formula 21b (S-isomer). Either the compound of Formula 21a (R-isomer) or Formula 21b (S- isomer) is reacted with CAD in the presence of a coupling reagent such as, for example, DCC and HOBT and the like in a solvent such as, for example,
CH2Cl2, CHCl3, THF and the like, optionally an
aprotic solvent such as, for example, DMF, DMSO and the like is added to aid solution, at about 0°C to about 25°C for about 4 hours to about 3 days to give the compound of Formula 22a (R-isomer) or Formula 22b (S-isomer). Either the compound of Formula 22a
(R-isomer) or Formula 22b (S-isomer) is treated with hydrogen in the presence of a catalyst such as, for example, 5% to 20% palladium on carbon, platinum oxide, 5% to 20% platinum on carbon, platinum oxide, 5% to 20% platinum on carbon, and the like in a solvent such as, for example, EtOH and the like to give the compound of Formula 23a (R-isomer) or
Formula 23b (S-isomer). Either the compound of
Formula 23a (R-isomer) or Formula 23b (S-isomer) is treated with an acid such as, for example,
trifluoroacetic acid, a nonpolar inert solvent such as, for example, CH2Cl2 and the like saturated with hydrogen chloride gas to give the compound of
Formula 24a (R-isomer) or Formula 24b (S-isomer) as the diacid addition salt. The diacid addition salt of the compound of Formula 24a (R-isomer) or
Formula 24b (S-isomer) may be converted to other pharmaceutically acceptable acid addition salts by conventional methodology as previously described.
The configuration of each isomer at the carbon atom to which the hydroxyl or derivatized hydroxyl group is attached is assigned as outlined in Scheme 3.
Thus, either the compound of Formula 19a or
Formula 19b is reacted with 2,2-dimethoxypropane in the presence of an acid catalyst such as, for
example, para-toluenesulfonic acid and the like followed by reduction with hydrogen in the presence of a catalyst such as, for example, rhodium on carbon and the like to afford the compound of Formula 25a or Formula 25b. 1H-NMR decoupling determinations indicate the stereochemistry of the protons attached to the five-membered ring.
Compounds of Formula I designated 30a and 30b are prepared, respectively, from a compound of
Formula 19a and 19b as outlined in Scheme 4 using the methodology used to prepare compounds of Formula 24a and 24b from compounds of Formula 19a and 19b as described in Scheme 2.
A compound of Formula I designated as 40 is prepared from a compound of Formula 31 as outlined in Scheme 5.
A compound of Formula I designated as 43 is prepared from a compound of Formula 32 as outlined in Scheme 6.
Compounds of Formula I designated 54 and 58 are prepared respectively from a compound of Formula 49 and a compound of Formula 44 as outlined in Scheme 7.
The preparation of selected parent renin
inhibitors of compounds of Formula I designated as 65a, 65b, 71, and 73 are as outlined in Schemes 8 and 9.
The compounds of the present invention can be prepared and administered in a wide variety of oral and parenteral dosage forms. It will be obvious to those skilled in the art that the following dosage forms may comprise as the active component, either a compound of Formula I or a corresponding
pharmaceutically acceptable salt of a compound of Formula I.
For preparing pharmaceutical compositions from the compounds of the present invention,
pharmaceutically acceptable carriers can be either solid or liquid. Solid form preparations include powders, tablets, pills, capsules, cachets,
suppositories, and dispersible granules. A solid carrier can be one or more substances which may also act as diluents, flavoring agents, binders,
preservatives, tablet disintegrating agents, or an encapsulating material.
In powders, the carrier is a finely divided solid which is in a mixture with the finely divided active component.
In tablets, the active component is mixed with the carrier having the necessary binding properties in suitable proportions and compacted in the shape and size desired.
The powders and tablets preferably contain from five or ten to about seventy percent of the active compound. Suitable carriers are magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth,
methylcellulose, sodium carboxymethylcellulose, a low melting wax, cocoa butter, and the like. The term "preparation" is intended to include the formulation of the active compound with encapsulating material as a carrier providing a capsule in which the active component with or without other carriers, is
surrounded by a carrier, which is thus in association with it. Similarly, cachets and lozenges are
included. Tablets, powders, capsules, pills,
cachets, and lozenges can be used as solid dosage forms suitable for oral administration.
For preparing suppositories, a low melting wax, such as a mixture of fatty acid glycerides or cocoa butter, is first melted and the active component is dispersed homogeneously therein, as by stirring. The molten homogenous mixture is then poured into
convenient sized molds, allowed to cool, and thereby to solidify.
Liquid form preparations include solutions, suspensions, and emulsions, for example, water or water propylene glycol solutions. For parenteral injection liquid preparations can be formulated in solution in aqueous polyethylene glycol solution.
Aqueous solutions suitable for oral use can be prepared by dissolving the active component in water and adding suitable colorants, flavors, stabilizing and thickening agents as desired.
Aqueous suspensions suitable for oral use can be made by dispersing the finely divided active
component in water with viscous material, such as natural or synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose, and other well-known suspending agents.
Also included are solid form preparations which are intended to be converted, shortly before use, to liquid form preparations for oral administration.
Such liquid foirms include solutions, suspensions, and emulsions. These preparations may contain, in addition to the active component, colorants, flavors, stabilizers, buffers, artificial and natural
sweeteners, dispersants, thickeners, solubilizing agents, and the like. The pharmaceutical preparation is preferably in unit dosage form. In such form the preparation is subdivided into unit doses containing appropriate quantities of the active component. The unit dosage form can be a packaged preparation, the package containing discrete quantities of preparation, such as packeted tablets, capsules, and powders in vials or ampoules. Also, the unit dosage form can be a capsules, tablet, cachet, or lozenge itself, or it can be the appropriate number of any of these in packaged form.
The quantity of active component in a unit dose preparation may be varied or adjusted from 0.1 mg to 2000 mg preferably 0.5 mg to 1000 mg according to the particular application and the potency of the active component. The composition can, if desired, also contain other compatible therapeutic agents.
In therapeutic use as agents for treating hypertension, hyperaldosteronism, congestive heart failure, and diseases caused by retroviruses
including HTLV I, II, and III as well as diagnostic tools for the identification of cases of hypertension due to renin excess, the compounds utilized in the pharmaceutical method of this invention are
administered at the initial dosage of about 0.1 mg to about 50 mg per kilogram daily. A daily dose range of about 0.5 mg to about 30 mg per kilogram is preferred. The dosages, however, may be varied depending upon the requirements of the patient, the severity of the condition being treated, and the compound being employed. Determination of the proper dosage for a particular situation is within the skill of the art. Generally, treatment is initiated with smaller dosages which are less than the optimum dose of the compound. Thereafter, the dosage is increased by small increments until the optimum effect under the circumstances is reached. For convenience, the total daily dosage may be divided and administered in portions during the day, if desired.
The following nonlimiting examples illustrate the inventors' preferred methods for preparing the compounds of the invention.
EXAMPLE 1
SMO-Phe-Ser(Lys•2HCl)-CAD
Step A: Preparation of SMO-Phe-Ser (Lys•2 Boc)-CAD Di-t-Boc-Lys (1.11 g, 3.19 mmol, 2.0 eq) in THF
(7 mL) is added to CDI (570 mg, 3.51 mmol, 2.2 eq) in THF (7 mL) at 0°C under N2 and stirred for
30 minutes. SMO-Phe-Ser-CAD (Example C) (1.0 g, 1.60 mmol) in THF (5 mL) is added and the mixture stirred at room temperature for 3 days. The solvents are evaporated and the residue partitioned between ethyl acetate (50 mL) and 2N HCl (50 mL). The organic layer is separated, washed with 2N sodium carbonate (50 mL), brine (50 mL), dried (MgSO4), filtered and evaporated to afford a white foam. This is purified by column chromatography on silica gel eluting with 2% then 3% MeOH/CH2Cl2 to give a white foam. This is further purified by column
chromatography on silica gel (thin layer
chromatography - TLC grade) eluting with 3%
methanol/dichloromethane to give 699 mg of a white foam.
1H NMR (300 MHz, d6DMSO + 2 drops D2O) δ 8.56 (1H, d,
J = 9Hz), 7.69 (1H, d, J = 10 Hz), 7.10-7.40
(6 H, m), 6.72 (1H, br t), 4.61 (1H, m), 4.10-4.30 (3H, m), 3.95 (2H, m), 3.35 (4H, m), 3.41 (1H, t, J = 9 Hz),, 2.80-3.00 (4H, m), 2.55-2.75 (5 H, m), 1.00-1.80 (40H, m), 0.86 (3H, d, J = 7.2 Hz), 0.78 (3H, d, J = 7.0 Hz). Step B: Preparation of SMO-Phe-Ser (Lys•2 HCl)-CAD
SMO-Phe-Ser(Lys•2Boc)-CAD (600 mg, 0.628 mmol) in CH2Cl2 (5 mL) is added to CH2Cl2 (20 mL) saturated with HCl (gas) at 0°C and stirred at 0°C for 2 hours. The solvents are evaporated and the residue taken up in water (30 mL) and washed with chloroform
(3 x 20 mL). The water layer is filtered through 'hyflo' and freeze-dried to give 425 mg of a white powder. Mass spectrum-fast atom bombardment (MS (FAB)) 755.2 (100%) - free base M+
EXAMPLE 2
SMO-Phe-Ser(Asp•HCl)-CAD Step A: Preparation of SMO-Phe-Ser(Asp•Boct-Bu)-CAD γ-t-Bu(N-Boc) aspartate dicyclohexylammonium salt (1.50 g, 3.19 mmol, 4 eq) is added to 5% citric acid (100 mL) and extracted with ethyl acetate
(3 x 50 mL), the extracts are washed with brine
(100 mL), dried (MgSO4), filtered and evaporated to an oil. This is dissolved in THF (5 mL) and added to CDI (569 mg, 3.51 mmol, 4.4 eq) at 0°C and stirred for 1 hour. Smo-Phe-Ser-CAD (Example C) (0.5 g, 0.80 mmol) in THF (2 mL) is added and the mixture stirred for 1 week. The mixture is diluted with
EtOAc (30 mL), washed with 5% citric acid solution (50 mL), 2 N Na2CO3 (50 mL), brine (50 mL), dried (MgSO4), filtered and evaporated to leave a brown solid. This is purified by column chromatography (2% to 3% MeOH/CH2Cl2) to afford the product as white foam (573 mg). Further chromatography (2%
MeOH/CH2Cl2, TLC silica) affords 277 mg of a white solid. 1H NMR (300 MHz, d6-DMSO + 2 drops D2O) δ 7.20-7.40 (6H, m), 4.62 (1H, t, J = 5.8 Hz),
4.40 (1H, dd, J= 9.7, 3.5 Hz), 4.10-4.30 (3H, m), 3,94 (1H, dd, J = 11.2, 3,3 Hz), 3.34 (4H, br s), 3.06 (1H, t, J = 5.2 Hz), 2.85-3.00 (2H, m), 2.40- 2 . 75 ( 8H, m) , 1 . 00-1. 80 (34H, m) , 0 . 84 (3H, d,
J = 6. 7 Hz ) , 0 . 78 (3H, d, J = 6. 4 Hz) .
Step B: Preparation of SMO-Phe-Ser(Asp•HCl)-CAD
Hydrogen chloride gas is passed over the
aspartate (588 mg, 0.65 mmol) in CH2Cl2 (20 mL) at room temperature and stirred for 1 hour. The solvent is evaporated and the residue dissolved in CHCl3.
This is evaporated to leave a white solid. This is dissolved in water (150 mL) and washed with EtOAc
(100 mL). Hexane (50 mL) is added to break down the emulsion that forms and the aqueous layer filtered through celite and evaporated to afford 447 mg of a white solid. 1H NMR (200 MHz, d6-DMSO + 2 drops D2O) 7.20-7.40 (6H, m), 4.91 (1H, t, J = 5.0 Hz), 4.82 (1H, d, J = 6.0 Hz), 4.46 (2H, m), 4.12 (1H, brt, J = 5.0 Hz), 3.69 (1H, brm), 3.34 (4H, br s), 2.50- 3.10 (11H, m), 1.00-1.80 (15H, m), 0.95 (3H, d, J = 7.0 Hz).
EXAMPLE 3
SMO-Phe-Thr(Lys•2HCl)-CAD
Step A: Preparation of SMO-Phe-Thr(Lys•2Boc)-CAD
Di-t-Boc-Lys (5.41 g, 15.6 mmol, 5 eq) in THF
(20 mL) is added to CDI (3.04 g, 18.7 mmol, 6 eq) in THF (20 mL) at 0°C under N2. After stirring at 0°C for 5 min, the solution becomes homogenous. TLC confirmed no lysine present. SMO-Phe-Thr-CAD
(Example D) (1.926 g, 3.00 mmol) in THF (20 mL) and DMAP (100 mg) are added at 0°C. The mixture is stirred at room temperature overnight and then at reflux for 24 hours. The cooled mixture is
evaporated, the residue taken up in EtOAc (300 mL), washed with 2 N HCl (200 mL), 2 N Na2CO3 (200 mL), brine (200 mL), dried (MgSO4), filtered and
evaporated to leave a white foam (~8 g). This is purified by column chromatography on silica gel (flash) eluting with 2% then 3% MeOH/CH2Cl2 to give a white foam (2.93 g). This is further purified by column chromatography twice on silica gel (2 x 100 g TLC grade) to afford 578 mg of a white foam. HPLC 98.6% pure.
Step B: Preparation of SMO-Phe-Thr(Lys•2HCl)-CAD
The lysine ester (600 mg) in CH2Cl2 (5 mL) is added to saturated HCl/CH2Cl2 (20 mL) at 0°C and the mixture stirred at room temperature for 4 hours. The solvent is evaporated to leave the product as an off- white solid. This is dissolved in water (50 mL) and washed with EtOAc (50 mL), filtered through celite and freeze-dried to afford 454 mg of a fluffy white solid. MS (FAB) 769.5 (17%) M+ of free base.
EXAMPLE 4
SMO-Phe-Ser(Glu•TFA)-CAD
Step A: Preparation of SMO-Phe-Ser(Glu•Cbz)-CAD
Figure imgf000072_0001
OBz
A mixture of SMO-Phe-Ser-CAD (Example C)
(300 mg, 0.479 mmol), Glu•Cbz•Bz (178 mg,
0.479 mmol), DDC (99 mg, 0.479 mmol), and DMAP
(12 mg, 95.7 μmol) in dichloromethane (5 mL) is stirred at room temperature for 16 hours. The mixture is filtered and evaporated to leave an oil. The oil is purified by column chromatography on silica gel (50 g TLC grade) eluting with 2% methanol/ dichloromethane to give 347 mg of a white solid; MS (FAB) 980.6 (42%) M+.
Step B: Preparation of SMO-Phe-Ser(Glu•TFA)-CAD
SMO-Phe-Ser(Glu•Cbz)-CAD (340 mg, 0.347 mmol) is
OBz
stirred in ethanol (20 mL) with 5% palladium on carbon (120 mg) and TFA (134 μL, 1.73 mmol) under hydrogen. After 6 hours the mixture is filtered through hyflo and evaporated to leave an oil. This oil is dissolved in water (50 mL). and ethanol
(30 mL), filtered through hyflo, and evaporated to remove the ethanol. The remaining aqueous phase is freeze-dried to give a white solid. This solid is dried to 50°C under high vacuum to give 277 mg of a white solid; MS (FAB) 756.5 (100%) M+. EXAMPLE 5
SMO-Phe-Ser(Gln•TFA)-CAD
Step A: Preparation of SMO-Phe-Ser(Gln•Cbz)-CAD
SMO-Phe-Ser-CAD (Example C) (300 mg, 0.479 mmol) is reacted with Gln•Cbz (134 mg, 0.479 mmol) under the same conditions in Example 4, Step A to give 392 mg of a colorless oil;
1H NMR (300 MHz, d6-DMSO + two drops D2O) :
δ 7.15-7.50 (11H, m), 5.01 (2H, s), 4.65 (1H, t, J = 6.1 Hz), 3.90-4.35 (5H, m), 3.35 (4H, s), 3.11 (1H, t, J = 9.2 Hz), 2.95 (2H, m), 2.15 (2H, t,
J = 7.3 Hz), 0.95-2.05 (18H, m), 0.83 (3H, d,
J = 6.6 Hz), 0.78 (3H, d, J = 6.2 Hz).
Step B: Preparation of SMO-Phe-Ser(Gln•TFA)-CAD
SMO-Phe-Ser(Gln•Cbz)-CAD (305 mg, 0.343 mmol) is reacted under the same conditions in Example 4, Step B to give 252 mg of a white powder;
1H NMR (300 MHz, d6-DMSO + two drops D2O) : δ 8.67 (1H, d, J = 7.9 Hz), 7.72 (1H, d, J. = 9.2 Hz),
7.15-7.40 (6H, m), 4.72 (1H, q, J = 6.7 Hz),
3.85-4.40 (5H, m), 3.34 (4H, s), 2.95-3.15 (3H, m), 2.45-2.75 (6H, m), 2.27 (2H, m), 2.02 (2H, m),
0.95-1.85 (15H, m), 0.86 (3H, d, J = 6.6 Hz), 0.79 (3H, d, J = 6.6 Hz). EXAMPLE 6
SMO-Phe-Ser(Ala•TFA)-CAD
Step A: Preparation of SMO-Phe-Ser(Ala-Cbz)-CAD
A mixture of SMO-Phe-Ser-CAD (Example C)
(300 mg, 0.468 mmol), Cbz-Ala (105 mg, 0.468 mmol, 1.0 eq), DCC (97 mg, 0.468 mmol, 1.0 eq) and DMAP (11 mg, 94 μmol, 0.2 eq) in dichloromethane (10 mL) is stirred at room temperature for 16 hours. The mixture is filtered and evaporated. The residue is purified by column chromatography on silica gel (50 g TLC grade) eluting with 2% methanol in
dichloromethane to give 407 mg of a colorless oil; MS (FAB) 832.2 (36%) M+. Step B: Preparation of SMO-Phe-Ser(Ala•TFA)-CAD
A mixture of SMO-Phe-Ser(Ala-Cbz)-CAD (349 mg, 0.419 mmol), 5% palladium on carbon (100 mg) and trifluoroacetic acid (162 μL, 5 eq) in ethanol
(50 mL) is stirred under hydrogen for 4 hours. The mixture is filtered through 'Hyflo' (diatomaceous earth) and evaporated to a white residue. This is dissolved in 1:1 ethanolrwater (100 mL), filtered through 'Hyflo' and evaporated until all of the ethanol has gone. The residue is freeze-dried to afford a white powder (292 mg); MS (FAB) 698.2 (100%) M+ of free base.
EXAMPLE 7
SMO-Phe-Mal-CAD(2•COCH2CH2CO2H) A mixture of SMO-Phe-Mal-CAD (U.S. 5,036,053)
(200 mg, 0.305 mmol), succinic anhydride (153 mg, 1.53 mmol, 5 eq) and DMAP (75 mg, 0.611 mmol, 2 eq) is stirred in CH2Cl2 (2.5 mL) at room temperature for 2 days. The solvent is evaporated and the residue taken up in EtOAC (30 mL) and washed with 2 N HCl (2 x 20 mL). The EtOAc layer is washed with brine (30 mL), dried (MgSO4), filtered and evaporated to leave a yellow foam. This is purified by column chromatography on silica gel eluting with 5%
MeOH/0.5% HOAc/CH2Cl2 to afford 2.04 mg of a white foam.
TLC silica gel plates (5% methanol/
dichloromethane/1% HOAc) Rf 0.39; detection UV and phosphomolybdic acid. 1H NMR (300 MHz, d6-DMSO) δ 12•15 (2H, brs), 8.87 (1H, 2 x d, J - 10.0 Hz),
8.31 (1H, d, J = 10.0 Hz), 7.73 (1H, t, J = 11.0 Hz), 7.15-7.50 (5H, m), 5.20 (1H, t, J = 9.0 Hz),
4.95 (2H, m), 4.20 (2H, m), 3.73 (3H, 2 x s),
3.03 (1H, m), 2.91 (1H, m), 2.35-2.75 (13H, m),
0.6-1.80 (21H, m). EXAMPLE 8
(R isomer at *)
Figure imgf000075_0001
Step A: Preparation of S
Figure imgf000075_0002
(R isomer at *)
Alpha-Boc,epsilon-Cbz-Lys, 2.15 g (5.64 mmol), is dissolved in 75 mL dry dichloromethane and cooled to 5°C. CDI (1.05 g, 6.49 mmol), is added, and the mixture is warmed to 25ºC over 1 hour. (Example I)
Figure imgf000075_0003
(R isomer at *), (2.0 g, 5.64 mmol) is dissolved in dichloromethane (50 mL), and is added to the
previously prepared solution. A trace of DMAP is added, and the mixture is stirred at 25°C for
2 weeks. The mixture is then consecutively washed with 1N citric acid, saturated NaCl solution,
saturated NaHCO3 solution and saturated NaCl
solution. The organic phase is dried over MgSO4, filtered and evaporated under reduced pressure to afford a glassy solid, 4.02 g. Chromatography of the solid on silica gel eluting with EtOAc/hexane (25/75) gives the R isomer product as a white foam, 2.88 g. The structure is confirmed by NMR and Mass
Spectroscopy; MS (FAB) 617.4 (100%) M+-Boc.
2
In a similar manner, S -CH2-CH=CH2 (S
Figure imgf000076_0001
isomer at *) (2.0 g, 5.64 mmol) gives the desired S isomer product as a white foam, 2.65 g. The
structure is confirmed by NMR and Mass Spectroscopy; MS (FAB) 617.4 (100%) M+-Boc. 2
Step B: Preparation of
(R isomer at *)
Figure imgf000076_0002
2
(R isomer at *)
Figure imgf000076_0003
(2.66 g, 3.71 mmol) is dissolved in 100 mL acetone and cooled to 3°C. In a separate flask, NaIO4
(5.52 g, 25.8 mmol) and ruthenium (IV) oxide hydrate (0.168 g) are dissolved in 65 mL H2O. The solution is filtered and added to the previous solution, giving an exotherm to 28°C. Temperature is
maintained by cooling at 25°C for 1.5 hours, after which isopropanol (5 mL) is added. After stirring for 15 minutes, the mixture is filtered, evaporated to an oily aqueous suspension, diluted with saturated NaCl solution and extracted into CHCl3. The organic extract is washed with a dilute Na2SO3 solution and the pH adjusted to 2.0 with 1 N HCl. The organic phase is washed with saturated NaCl solution, dried over MgSO4, filtered and evaporated to give the R isomer product as a white foam, 2.08 g. The
structure is confirmed by NMR and Mass Spectroscopy; MS (FAB) 735.3 (4.65%) M+.
Similar treatment of
,
Figure imgf000077_0001
(S isomer at *), 2.48 g (3.46 mmol) gives the
corresponding S isomer product as a white foam,
2.11 g. The structure is confirmed by NMR and Mass Spectroscopy; MS (FAB) 735.5 (3.8%) M+.
Step C: Preparation of
(R isomer at *)
Figure imgf000077_0002
(R isomer at *) (1.90 g,
Figure imgf000077_0003
2.58 mmol) is dissolved in dichloromethane (100 mL). A solution of HOBT (0.36 g, 2.66 mmol) in 5 mL DMF is added followed by a solution of CAD (0.63 g,
2.58 mmol in dichloromethane (25 mL). The mixture is stirred and allowed to warm to 25°C overnight. The mixture is filtered and evaporated under reduced pressure. The residue is taken up into EtOAc and washed consecutively with solutions of saturated NaCl, 1 N citric acid, saturated NaHCO3, and
saturated NaCl. The organic phase is dried over MgSO4, filtered, and evaporated to a foam, 2.61 g. Chromatography on silica gel eluting with
EtOAc/Hexane (40/60) gives the R isomer product as a white foam, 1.57 g. The structure is confirmed by NMR and Mass Spectroscopy. MS (FAB) 960.8 (14.3%) M+.
2
In an identical manner, S _ o o
Figure imgf000078_0001
(S isomer at *) (1.90 g, 2.58 mmol), yielded 1.72 g of the corresponding S isomer product as a white foam. The structure is confirmed by NMR and Mass Spectroscopy; MS (FAB) 960.6 (15.1%) M+.
Step D: Preparation of S
Figure imgf000078_0002
(R isomer at *) (R isomer at *) (1.45 g,
Figure imgf000078_0003
1.51 mmol) is dissolved in methanol (100 mL) and 20% palladium on carbon catalyst (0.25 g) is added. The suspension is purged with H2 gas for 3 hours, filtered, and evaporated under pressure to a white foam. The foam is dissolved in a minimal amount of dichloromethane and the resulting solution is added to Et2O giving a solid precipitate. The solid is filtered, washed with Et2O and dried in vacuo to give 0.94 g of the R isomer product as a white solid. The structure is confirmed by NMR and Mass Spectroscopy; MS (FAB) 826.7 (100%) M+. 2
In an identical manner,
Figure imgf000079_0001
(S isomer at *) (1.60 g, 1.67 mmol) gives the S isomer product as a white solid, 1.07 g. The
structure is confirmed by NMR and Mass Spectroscopy; MS (FAB) 826.7 (100%) M+. 2
Step E: Preparation of (R isomer at *)
Figure imgf000079_0002
2
(R isomer at *) (0.92 g,
Figure imgf000079_0003
1.11 mmol) is dissolved in 25 mL dichloromethane and occasionally purged with anhydrous HCl gas. After 1 hour, the mixture is evaporated under reduced pressure to a white solid. The solid is redissolved in dichloromethane, filtered, and the filtrate is added to Et2O giving a solid precipitate. The solid is filtered, washed with Et2O and dried under reduced pressure giving the R isomer product as a white solid, 0.807 g. The structure is confirmed by NMR and Mass Spectroscopy; MS (FAB) 726.4 (100%) M+.
In a similar manner,
Figure imgf000079_0004
(S isomer at *) (1.05 g, 1.27 mmol) gives the S isomer product as a white solid, 0.884 g. The structure is confirmed by NMR and Mass Spectroscopy; MS (FAB) 726.3 (100%) M+.
EXAMPLE 9
2
2 2 2
Figure imgf000080_0001
Step A: Preparation of
Figure imgf000080_0002
(R isomer at *)
-A 2.52 g (7.05 mmol) is dissolved in
Figure imgf000080_0003
z
125 mL CH2Cl2, and CDI, 1.31 g (8.11 mmol) is added and stirred for 5 minutes at room temperature.
2
Figure imgf000080_0004
(Example I) (R Isomer at *) 2.50 g (7.05 mmol) is added, and the mixture is stirred at room temperature for 7 days. The mixture is evaporated under reduced pressure to an oil, resuspended in Et2O, extracted with 1N citric acid, saturated NaCl solution,
saturated NaHCO3, and saturated NaCl solution. The organic phase is dried over MgSO4, filtered, and evaporated to a crude gum, 4.75 g. Chromatography on silica gel, eluting with EtOAc/Hexane (20/80) gives the product as a syrup, 4.41 g. NMR spectroscopy confirms the structure. Mass Spectroscopy further confirms the structure; MS (FAB) 694.0 (65.2%) M+. Step B: Preparation of S O O
Figure imgf000081_0001
(R isomer at *) 2
O -CH=CH2, 4.31 g (6.21 mmol)
O
(R isomer at *)
Figure imgf000081_0002
is dissolved in 170 mL acetone and cooled to 3°C. A solution of NaIO4 9.24 g (43.2 mmol) and RuO2xH2O 0.28 g in 130 mL H2O is added to the preceding solution, giving an exotherm which is controlled to 20°C by external cooling. After stirring at room temperature for 2 hours, the suspension is filtered through celite and evaporated under reduced pressure to remove acetone. The residue is extracted twice with Et2O, and the extract is then washed with saturated NaCl solution, 5% sodium bisulfite
solution, and saturated NaCl solution. The organic phase is dried over MgSO4, filtered, and evaporated to a solid 3.65 g. NMR spectroscopy confirms the structure. Mass. Spectroscopy further confirms the structure; MS (FAB) 712.0 (46.6%) M+.
2
Step C: Preparation of
Figure imgf000082_0001
OBz
(R isomer at *)
2
-CO2H, 3.52 g (4.95 mmol) is
O )
(R isomer at *)
Figure imgf000082_0002
dissolved in 175 mL CH2Cl2, and cooled to 0°C. A solution of HOBT 0.7 g (5.19 mmol) in DMF is added, followed by DDC 1.07 g (5.19 mmol), and a solution of
CAD 1.20 g (4.94 mmol) in 20 mL CH2Cl2/DMF. The mixture is stirred at room temperature overnight and is evaporated under reduced pressure to an oil with suspended solids. The mixture is suspended in Et2O, filtered, and extracted with 1N citric acid,
saturated NaCl solution, saturated NaHCO3, and saturated NaCl solution. The organic phase is dried over MgSO4, filtered, and evaporated to a white foam, 4.72 g. Chromatography on silica gel, eluting with a gradient of 25% to 50% EtOAc in Hexane gives the product as a solid, 3.43 g. NMR spectroscopy
confirms the structure. Mass Spectroscopy further confirms the structure; MS (FAB) 937.4 (42.0%) M+.
Step D: Preparation of
Figure imgf000083_0003
(R isomer at *)
S 3.44 g (3.63 mmol) (R isomer at *)
Figure imgf000083_0002
is dissolved in THF, 20% Pd on carbon catalyst is added, and the suspension is pressurized to 50 psi with H2 gas. After 18 hours the mixture is vented, filtered, and the filtrate is evaporated under reduced pressure to a grey foam. Chromatography on silica gel, eluting with MeOH/CHCl3 (20/80) gives the product as a grey solid. The solid is dissolved in
CH2Cl2, filtered through charcoal, and a solid is precipitated from the filtrate by addition of Et2O.
The solid is filtered, washed with Et2O, and dried under reduced pressure to a white solid 0.88 g. NMR spectroscopy confirms the structure. Mass
Spectroscopy further confirms the structure; MS (FAB)
713.4 (73.5%) M+.
2
In a similar manner, O
Figure imgf000083_0001
(R isomer at *)
gives the S isomer. EXAMPLE 10
(Lys)O-CH2- -Phe-Alg-CAD•2HCl
Figure imgf000084_0001
Step A: Preparation of (Lys-2Boc)O-CH2
Figure imgf000084_0002
-Phe-Alg-CAD
(Lys-2Boc)O-CH2-CO2H (Example Q) (1.30 g,
3.2 mmol) is dissolved in 10 mL of CH2Cl2 and cooled to 0°C, HOBT (433 mg, 3.2 mmol), DCC (661 mg,
3.2 mmol) and DMAP (195 mg, 1.60 mmol) are added sequentially. The white suspension is stirred for 15 minutes. Phe-Alg-CAD (Example N) (780 mg,
1.60 mmol) in 10 mL of dry CH2Cl2 is added dropwise and the reaction mixture is stirred at room
temperature for 3 days. The reaction is filtered through Celite. The filtrate is washed with 1N HCl, saturated NaHCO3, and brine. The organic phase is dried with MgSO4 and concentrated under vacuum. The crude product is purified twice by silica gel
chromatography with 1% MeOH/CH2Cl2 as eluent to give 610 mg of pure product; MS (FAB) 874 (M+), 774, 674 (100%).
Step B: Preparation of (Lys•2HCl)O-CH2
Figure imgf000084_0003
-Phe-Alg-CAD
(Lys-2Boc)O-CH2-
Figure imgf000084_0004
-Phe-Alg-CAD (100 mg,
0.11 mmol) is dissolved in 10 mL of dry CH2Cl2 and cooled to 0°C. Hydrogen chloride gas is passed through the reaction flask for 10 minutes and
stirring is continued for 1 hour. A white
precipitate appears and solvent is evaporated and the jelly residue is dissolved in CH2Cl2 and evaporated to dryness. The process is repeated several times to afford a white solid. It is washed with EtOAc/Et2O (1:2) and collected as fine white powder (58 mg). HPLC showed 92% purity; MS (FAB) 675 (100% M+ free base) 675 (M+-H2O).
EXAMPLE 11
(Asp)O-CH2-
Figure imgf000085_0001
-Phe-Alg-CAD•HCl Step A: Preparation of (Asp•Boc•t-Bu)O-CH2
Figure imgf000085_0002
-Phe- Alg-CAD
The (N-Boc-α-t-butylaspartyl) glycolate (1.01 g, 2.9 mmol) is dissolved in 10 mL of CH2Cl2. DCC
(600 mg, 2.9 mmol), HOBT (393 mg, 2.9 mmol) and DMAP (178 mg, 1.46 mmol) are added sequentially to the reaction vessel at 0°C. After 15 minutes Phe-Alg-CAD (Example N) (850 mg, 1.74 mmol) in 10 mL of CH2Cl2 is added. The reaction mixture is stirred at room temperature for 2 days and worked up as usual.
Silica gel chromatography with 1% to 2% MeOH/CH2Cl2 affords 760 mg of product as a white foam; MS (FAB) 818 (M+ 48%), 762, 744, 718, 688, 644.
Step B: Preparation of (Asp•HCl)O-CH,
Figure imgf000085_0003
-Phe-Alg- CAD•HCl
Figure imgf000085_0004
(Asp•Boc•t-Bu)O-CH2-C-Phe-Alg-CAD (201 mg,
0.25 mmol) is dissolved in 20 mL of CH2Cl2. Hydrogen chloride gas is passed through the reaction mixture at 0°C for 15 minutes. The reaction is continuously stirred for 3 hours at room temperature. Solvent is removed and the residue is dissolved in fresh CH2Cl2 and evaporated. The process is repeated several times to afford a white solid which is washed with Et2O/EtOAc and collected as a fine white powder
(159 mg); MS (FAB) 661 (M+ 91%) 643. EXAMPLE 12
( O -Phe-Atm-CAD•HCl
Figure imgf000086_0001
Step A: Preparation of (Asp•Boc•t-Bu) OCH2 -Phe-Atm-CAD
Figure imgf000086_0002
(Asp•Boc•t-Bu) -Phe-Atm(Cbz)-CAD
Figure imgf000086_0003
(373 mg, 0.35 mmol) (Example U) is dissolved in 25 mL of MeOH and 200 mg of TsOH (3 eq) and a catalytic amount of 20% palladium on carbon is added. The mixture is stirred under one atmosphere of hydrogen for 6 hours, filtered, and the solution neutralized with solid NaHCO3. The MeOH is evaporated and the residue diluted with water and EtOAc. The mixture is extracted with EtOAc (3 x 30 mL) and dried (magnesium sulfate). The product is purified by chromatography with 3% MeOH:CH2Cl2 to afford 140 mg of product as a white foam; MS (FAB) 932 (M+1).
Step B: Preparation of -Phe-Atm-CAD•HCl
Figure imgf000086_0004
(Asp•Boc•t- Bu) O- -Phe-Atm-CAD (140 mg,
Figure imgf000086_0005
0.15 mmol) is dissolved in 10 mL of dry CH2Cl2, cooled to 0°C, and hydrogen chloride gas passed through for 1 hour, and the reaction stirred for 2 hours at room temperature. The solvent is evaporated and the residue treated with CH2Cl2 and evaporated. The yellow solid is collected by
filtration. The filter cake is washed many times with EtOAc and dried under vacuum; MS (FAB) 776 (M+1), 758 (M-18).
EXAMPLE 13
(Asp) O- -Phe-Alg-CAD
Figure imgf000087_0003
Step A: Preparation of
(Asp•Boc•t-Bu) 0- -Phe-Alg-CAD
Figure imgf000087_0002
(Asp•Boc•t-Bu)O- (1.55 g, 4.1 mmol)
(Example S, Step B ac
Figure imgf000087_0001
id before coupling to Phe-O-Bz is used) is dissolved in 70 mL of CH2Cl2 cooled to 0°C, DCC (933 mg, 4.52 mmol), HOBT (610 mg,
4.52 mmol), DMAP (250 mg, 2.05 mmol), and Phe-Alg-CAD (1.0 g, 2.01 mmol) (Example N) are added. The mixture is stirred at room temperature for 2 days, filtered, and the solution washed with IN HCl, saturated NaHCO3, NaCl, and dried (magnesium
sulfate). The product is purified by chromatography with 2% MeOH:CH2Cl2 to afford 1.3 g of the product as a white foam. MS (FAB) 859 (M+1). 3
Step B: Preparation of (Asp)O 2 -CAD
Figure imgf000088_0001
3
3
(Asp•Boc•t-Bu)O -Alg-CAD (196 mg,
Figure imgf000088_0002
0.23 mmol) is dissolved in 15 mL of CH2Cl2. Hydrogen chloride gas is passed through the solution for about
30 minutes at 0°C and the mixture stirred at 0°C to room temperature for a total of 3 hours. The CH2Cl2 is evaporated and the residue rinsed with CH2Cl2 several times and the white solid collected by filtration and washed with EtOAc/Et2O to afford
143 mg of the product as a white powder; MS (FAB)
702 (M+1) 685 (m-18+1).
PREPARATION OF STARTING MATERIALS
EXAMPLE A
Cbz-Ser-CAD
Cbz-Ser (500 mg, 2.09 mmol) is stirred in CH2Cl2 (30 mL) and DMF (10 mL) at 0°C and DCC (474 mg,
2.30 mmol, 1.1 eq), CAD (534 mg, 2.19 mmol, 1.05 eq) and HOBT (310 mg, 2.30 mmol, 1.1 eq) is added. The mixture is stirred for 2 days, filtered and
evaporated. The residue is taken up in ethyl acetate (50 mL) and washed with 2 N Na2CO3 (75 mL), brine (75 mL), dried (MgSO4), filtered and evaporated to afford an off-white solid. This is purified by column chromatography on silica gel eluting with 2%, 3% then 4% methanol/CH2Cl2 to give 600 mg of a white solid. A portion (100 mg) is recrystallized from chloroform to give 67 mg of a white solid;
mp 144-146°C. EXAMPLE B
Ser-CAD
Cbz-Ser-CAD (Example A) (6.8 g, 14.6 mmol) in EtOH (100 mL) with 20% palladium on carbon (0.5 g) is shaken under H2 at 50 pounds per square inch (psi) at room temperature for 1 hour. The mixture is filtered and evaporated to leave a white foam. This is recrystallized from ethyl acetate to give 4.03 g of a fluffy white solid. Additional product (406 mg) is obtained from the mother liquors on standing; MS (FAB) 331.2 (100%).
EXAMPLE C SMO-Phe-Ser-CAD
A mixture of SMO-Phe (European Published
Application EP 0399,556) (1.90 g, 6.06 mmol, 1.0 eq), Ser-CAD (Example B) (6.06 mmol, 2.0 g, 1.0 eq), DCC (1.37 g, 6. 66 mmol, 1.1 eq) and HOBT (0.90 g,
6.66 mmol, 1.1 eq) is stirred in CH2Cl2 (100 mL) and DMF (50 mL) at room temperature under N2 for 3 days. The solvents are evaporated and the residue taken up in ethyl acetate (200 mL). This is washed with 2 N Na2CO3 (200 mL), brine (200 mL), dried (MgSO4), filtered and evaporated to afford a white foam. This is purified by column chromatography on silica gel eluting with 3% then 4% MeOH/CH2Cl2 to give a white foam; mp 99-102°C.
EXAMPLE D SMO-Phe-Thr-CAD
Step A: Preparation of SMO-Phe-Thr
A mixture of SMO-Phe (European Published
Application EP 0399,556) (16.6 g, 53 mmol) and HOBT (7.83 g, 58 mmol, 1.1 eq) in dichloromethane (400 mL) and dimethylformamide (50 mL) is cooled to 0°C under nitrogen. Threonine benzyl ester, hydrochloride (13.6 g, 55 mmol, 1.05 eq), triethylamine (8.06 mL, 58 mmol, 1.1 eq) and DCC (12.03 g, 58 mmol, 1.1 eq) are added and the mixture stirred at 0°C for 1 hour. After warming to room temperature, the mixture is filtered and evaporated. The residue is partitioned between ethyl acetate and water, the organic layer separated, washed with 2N sodium bicarbonate, brine, dried (MgSO4), filtered, and evaporated. The residue is purified by column chromatography on silica gel eluting with dichloromethane then 4%
methanol/dichloromethane to afford 19 g of a solid. This is dissolved in methanol (300 mL) and 5%
palladium on carbon (1.9 g) added. The mixture is stirred vigorously under hydrogen at room temperature for 5 hours. The mixture is filtered and evaporated to leave 14.8 g of a white solid; 1H NMR (90 MHz, CDCl3 + d6-DMSO) δ 7.10-7.50 (5H, m), 6.29 (1H, d, J 10.2Hz), 5.49 (3H, br. s), 3.90-4.55 (3H, m),
3.30-3.50 (4H, m), 2.75-3.10 (6H, m) and 1.15 (3H, d, J 6.6Hz).
Step B: Preparation of SMO-Phe-Thr-CAD
SMO-Phe-Thr (3.98 g, 9.57 mmol) is stirred at 0°C in DMF (30 mL) and dichloromethane (120 mL) and DCC (2.17 g, 10.5 mmol, 1.1 eq), HOBT (1.42 g,
10.5 mmol, 1.1 eq) and CAD (2.44 g, 10.1 mmol,
1.05 eq) added. The mixture is stirred at room temperature under nitrogen for 2 days and filtered. The filtrate is washed with 2N sodium carbonate
(2 x 100 mL), brine (100 mL), dried (MgSO4),
filtered, and evaporated to leave an oily foam. This is purified by column chromatography on silica gel, eluting with 2% methanol/dichloromethane to give a white foam. This is crystallized from
dichloromethane to give 2.41 g of a fluffy white solid; MS (FAB) 641.6 (51.3%) (M+H)+. EXAMPLE E
SMO-Phe-N(CH3)OCH3
A solution of 8.0 g (25.4 mmol) of SMO-Phe
(European Published Application EP 0399,556) in
100 mL of THF/CH2Cl2 (1/1) is cooled to -40°C and
4.54 g (28 mmol) of carbonyldiimidazole added. The mixture is then kept at -5°C for 1.5 hours. To this is added a solution of 2.73 g (28 mmol) of
0,N-dimethylhydroxylamine, hydrochloride, and 3.23 mL (28 mmol) of N-methylpiperidine in 40 mL CH2Cl2.
After stirring at room temperature overnight, the mixture is filtered and the filtrate evaporated. The residue is taken up in EtOAc and washed with 1 N citric acid, saturated NaCl, saturated NaHCO3, and saturated NaCl. Drying over MgSO4 and removal of the solvent under reduced pressure leaves 9.69 g of the crude product. Chromatography on silica gel, eluting with EtOAc/CHCl3/MeOH (49/49/2) gives 7.56 g of the product as a syrup which solidifies. The structure is confirmed by NMR and mass spectroscopy; MS (FAB) 358.1 (100%) M+.
EXAMPLE F
Figure imgf000091_0001
Under nitrogen, a Grignard solution prepared from 5.44 g (224 mmol) of Mg turnings and 22.7 mL (224 mmol) of 4-bromo-1-butene in 350 mL THF is heated to reflux, then cooled to -5°C and treated with a suspension of 20.0 g (55.9 mmol) of SMO-Phe- N(CH3)OCH3 (Example E) in 135 mL THF. After stirring at room temperature overnight, the mixture is
evaporated to an oil. The oil is poured into a cold, saturated solution of NH4Cl and extracted with EtOAc. The EtOAc is washed with 1 N citric acid, saturaaed NaCl, saturated NaHCO3, and saturated NaCl. Drying over MgSO4 and removal of the solvent under reduced pressure gives the crude product as an oil.
Chromatography on silica gel, eluting with
hexane/EtOAc (70/30) gives 16.9 g of the product as an oil. The structure is confirmed by NMR and Mass Spectroscopy; MS (EI) 353.28 M+.
EXAMPLE G
2
Figure imgf000092_0001
A solution of 6.35 g (18.0 mmol) of
SMO-NH -CH=CH2 (Example F) in 200 mL of
Figure imgf000092_0002
absolute EtOH is treated with 3.89 g (72.0 mmol) of KBH4 followed by 20 mL of H2O. After stirring at room temperature for 2.5 hours, 100 mL of acetone is added and the mixture stirred for 15 minutes. The suspension is filtered and the filtrate evaporated to an oil which solidifies. There is obtained 6.04 g of the product. The structure is confirmed by NMR and Mass Spectroscopy; MS (EI) 355.25 (3.8%) M+. EXAMPLE H
Figure imgf000092_0003
A solution of 5.95 g (16.8 mmol) of
2
CH2-CH=SH2 (Example G) in 100 mL THF
Figure imgf000092_0004
is treated with 1.48 g (22 mmol) of imidazole and 3.29 g (22 mmol) of t-butyldimethylsilyl chloride and the mixture stirred at room temperature for 2 days. An additional 0.8 g (11.8 mmol) of imidazole and 1.77 g (11.7 mmol) of t-butyldimethylsilyl chloride is then added and the mixture stirred overnight. The solvent is removed under reduced pressure and the residue suspended in EtOAc/Et2O (1/1) and washed with H2O, 1 N citric acid, saturated NaCl, saturated
NaHCO3, and saturated NaCl. Drying over MgSO4 and removal of the solvent under reduced pressure gives the crude product as a mixture of diastereomers.
Chromatography on silica gel, eluting with a gradient of 0% to 20% EtOAc in hexane gives 3.34 g of the faster eluting diastereomer as a glass. The
structure is confirmed by NMR and Mass Spectroscopy; MS (EI) 469 (100%) M+.
Continued elution from the column gives 2.86 g of the slower eluting diastereomer as a glass. The structure is confirmed by NMR and Mass Spectroscopy; MS (EI) 469 (100%) M+.
EXAMPLE I
SMO-NH- 2-CH2-CH=CH2 (Separate isomers, R and
Figure imgf000093_0001
S at *)
SMO-NH -CH2-CH=CH2 (derived from the faster
Figure imgf000093_0002
eluting R isomer at *) (Example H) 9.02 g (18.5 mmol) is dissolved in 350 mL anhydrous THF to which a solution of tetrabutylammonium fluoride (1 M in THF), 80 mL is added. After stirring at 25°C for 3 hours, the mixture is evaporated under reduced pressure to an oil. The oil is suspended in Et2O and extracted with 1 N citric acid, saturated NaCl solution, saturated NaHCO3 solution and saturated NaCl
solution. The organic phase is dried over anhydrous MgSO4, filtered and evaporated to a crude oil, 9 g in weight. The oil is crystallized from a mixture of Et2O/hexane (10/90) giving a white solid, 5.55 g. The structure is confirmed by NMR and Mass
Spectroscopy; MS (FAB) 355.3 (100%) M+.
In an identical manner, 7.2 g (14.8 mmol) of
2
SMO-NH CH2-CH=CH2 (derived from the slower
Figure imgf000094_0001
eluting S isomer at *) (Example H) yields 4.79 g of the corresponding S isomer product as a white solid. The structure is confirmed by NMR and Mass
Spectroscopy; MS (FAB) 355.3 (100%) M+.
EXAMPLE J -CH2CH2CH, (Separate R and S isomers
Figure imgf000094_0002
at *) -CH2-CH=CH2 (Example I)
Figure imgf000094_0003
(R isomer at *), 0.50 g (1.41 mmol) is dissolved in 40 mL of 2,2-dimethoxypropane and heated to 50°C. A trace of anhydrous p-toluenesulfonic acid is added, and the mixture is stirred for 1.25 hours. The mixture is evaporated under reduced pressure and saturated NaHCO3 solution is added to the residue. The oily suspension is extracted into Et2O, washed with saturated NaCl solution and is evaporated to an oil. Chromatography on silica gel, eluting with Hexane/EtOAc (5/95) gives 0.35 g of an oil. The oil is dissolved in 75 mL isopropanol and 10% Rhodium on carbon catalyst, 0.5 g is added. The mixture is placed under H2 gas at 50 psi for 30 hours, filtered to remove the catalyst, and evaporated under reduced pressure. The residue is resuspended in
2,2-dimethoxypropane and a trace of p-toluenesulfonic acid is added. The mixture is heated to 50°C for 1 hour, and evaporated under reduced pressure. The residue is suspended in saturated NaHCO3 solution, extracted into Et2O, washed with saturated NaCl solution, and evaporated to an oil. Chromatography on silica gel, eluting with EtOAc/Hexane (25/75) gives the product as an oil, 0.18 g. NMR
spectroscopy confirms R stereochemistry at *, and Mass Spectroscopy further confirms the structure; MS (EI) 403 (17.5%) M+. In a similar manner, 0.50 g (1.41 mmoles) of
2
-CH2-CH=CH2 (S isomer at *) gives the
Figure imgf000095_0001
S isomer product as an oil, 0.18 g. NMR spectroscopy confirms the structure and the S stereochemistry at *. Mass Spectroscopy further confirms the structure; MS (EI) 403 (15.4%) M+.
EXAMPLE K
Boc-Alg-CAD
To a solution of Boc-Alg (7.36 g, 34.23 mmol) in 200 mL of dry CH2Cl2 at 0°C are added DCC (8.48 g, 41.10 mmol), DMAP (2.5 g, 20.49 mmol) and HOBT
(5.55 g, 41.07 mmol). The resulting white suspension is stirred for 15 minutes. CAD (8.32 g, 34.23 mmol) is dissolved in 100 mL of dry Et2O and added to the reaction mixture dropwise through an addition funnel. After stirring at room temperature overnight, the reaction mixture is filtered and the white solid washed with CH2Cl2 thoroughly. The filtrate is evaporated and the residue taken up in 500 mL of
EtOAc and washed with IN HCl, saturated NaHCO3, and dried (MgSO4). Solvent is removed and the residue purified by silica gel chromatography with 20%
EtOAc/hexane to give 13.0 g of the product as a white solid; MS (FAB) 441 (M+), 385, 341.
EXAMPLE L
Alg-CAD
Boc-Alg-CAD (Example K) (1.01 g, 2.29 mmol) is dissolved in 100 mL of dry CH2Cl2. Hydrogen chloride gas is passed through this solution for 10 minutes. The reaction mixture is stirred for 3 hours. Solvent is removed and the residue dissolved in H2O (50 mL). The aqueous solution is extracted with Et2O
(50 mL x 2). The aqueous layer is then basified to pH 14 with NaOH pellets. The cloudy solution is extracted with EtOAc/Et2O (100 mL x 3). The organic layer is dried with MgSO4. Solvent is removed under reduced pressure to afford a white solid (760 mg); 1H NMR (CDCl3 + D2O) 7.5 (br d, 1H), 5.7 (m, 1H),
5.15 (m, 2H), 4.31 (m, 1H), 3.6 (m, 1H), 3.2 (br m, 2H), 2.5 (m, 2H), 1.9 (m, 1H), 1.8-1.1 (m, 14H), 0.95 (d, 3H), 0.93 (d, 3H). EXAMPLE M
Boc-Phe-Alg-CAD
To a solution of Boc-Phe (5.8 g, 21.89 mmol) in 200 mL of CH2Cl2 at 0°C is added sequentially DMAP (1.12 g, 9.18 mmol), HOBT (3.7 g, 27.38 mmol) and DCC (5.27 g, 25.54 mmol). The resulting white suspension is stirred for 15 minutes. A solution of Alg-CAD (Example L) (6.2 g, 18.23 mmol) in 100 mL of CH2Cl2 is added to the reaction mixture through an addition funnel. The reaction mixture is stirred vigorously at room temperature overnight. The white solid is filtered and the filtrate washed with 1 N HCl, saturated NaHCO3, and brine. The organic solution is dried (MgSO4) and concentrated and further purified by silica gel chromatography to give a white solid (7.1 g); MS (FAB) 589 (M+ +1) 514. EXAMPLE N
Phe-Alg-CAD
Boc-Phe-Alg-CAD (Example M) (7.0 g, 11.92 mmol) is dissolved in 500 mL of CH2Cl2 with the aid of a few drops of MeOH. Dry HCl gas is passed through solution for 15 minutes and the reaction mixture is further stirred for another 3 hours. Solvent is evaporated and the residue dissolved in H2O (200 mL). It is then extracted with CH2Cl2 (3 x 250 mL). The aqueous layer is basified with NaOH pellets and extracted with CH2Cl2 (3 x 300 mL). The combined extracts are dried (MgSO4) and evaporated to give 5.70 g of white solid; MS (FAB) 489 (M+ +1) 471
(M+-H2O). EXAMPLE O
TBDMSOCH2-
Figure imgf000097_0001
-Phe-Alg-CAD
TBDMSOCH2CO2H (1.05 g, 5.53 mmol) is dissolved in 15 mL of dry CH2Cl2 at 0°C. HOBT (824 mg,
6.10 mmol), DMAP (338 mg, 2.72 mmol), and DCC
(11.26 g, 6.10 mmol) are added. After 15 minutes, a solution of Phe-Alg-CAD (Example N) in 15 mL of
CH2Cl2 is added slowly. Reaction mixture is stirred at room temperature overnight. Dicyclohexylurea is filtered and the filtrate washed with 1N HCl, saturated NaHCO3, and dried with MgSO4. The solvent is removed and the residue chromatographed with 2% MeOH/CH2Cl2 to give 1.0 g of product as a white foam; MS (FAB) 659 (M+) 602.
EXAMPLE P
HO-CH2-
Figure imgf000098_0001
-Phe-Alg-CAD TBDMSOCH2-
Figure imgf000098_0002
-Phe-Alg-CAD (Example 0) (700 mg,
1.06 mmol) is dissolved in 15 mL of MeOH. A
catalytic amount of para-toluenesulfonic acid is added and the reaction mixture heated to 78°C for 1 hour. The methanol is evaporated and the residue chromatographed over silica gel with 2% to 3%
MeOH/CH2Cl2 to give the product as a white foam
(325 mg). A portion of the product is recrystallized from MeOH/EtOAc/hexane to give 16 mg of white powder; mp 185-187°C; MS (FAB) 546 (M+), 528 (M+-H2O).
EXAMPLE Q
(Lys-2Boc)O-CH2-CO2H
Step A: Preparation of (Lys-2Boc) O-CH2-CO2Bz
Carbonyldiimidazole (2.25 g, 13.88 mmol) is added to the solution of Bis-Boc Lysine (4.00 g, 11.56 mmol) in 40 mL of THF at 0°C followed by a catalytic amount of DMAP. After 10 minutes, benzyl glycolate (1.28 g, 7.7 mmol) is added and the
solution refluxed overnight. The solvent is
evaporated and the residue taken up in 300 mL EtOAc. It is washed with 1N HCl and with saturated NaHCO3. The organic phase is dried with MgSO4 and
concentrated. The crude reaction product is
chromatographed with 15% EtOAc/hexane on silica gel to give 3.6 g of product as a colorless oil; MS (FAB) 495 (M+) 439, 395. Step B: Preparation of (Lys-2Boc)O-CH2-CO2H
(Lys-2Boc)O-CH2CO2Bz (4.90 g, 9.9 mmol) is dissolved in 100 mL of EtOAc and 0.5 g of 20%
palladium on carbon is added. The hydrogenolysis is carried out under 50 psi until all the starting material is consumed. The catalyst is filtered and washed thoroughly with EtOAc. Solvent is removed to leave a viscous gel (3.80 g); MS chemical ionization (CI), 408 (M++1).
EXAMPLE R
Figure imgf000099_0001
( o e at )
Step A: Preparation of
Figure imgf000099_0002
OTBDMS
(S isomer at *) 2.77 g (5.91 mmol) )
Figure imgf000099_0003
(Example H) is dissolved in 140 mL acetone and cooled to 15°C. NaIO4 9.50 g (44.41 mmol) and RuO2xH2O,
0.10 g are dissolved in 60 mL H2O and added to the previously described solution, giving an exotherm which is controlled at 20°C by external cooling over
2 hours. Isopropanol, 30 mL is added to the mixture followed by filtration through celite. The filtrate is evaporated under reduced pressure and the residue is saturated with solid NaHCO3. The mixture is extracted exhaustively with CHCl3 which is washed with 4% NaSO3 solution buffered to pH 2 with
concentrated HCl. The organic phase is washed with saturated NaCl solution, dried over MgSO4, filtered, and evaporated to a solid, 2.51 g. The solid is filtered through silica gel eluted with MeOH:CHCl3 (10:90). Recrystallization from hexane gives a solid, 1.95 g. NMR spectroscopy confirms the structure. Mass Spectroscopy further confirms the structure; MS (El+) 487.25 (100.0%) M+.
Step B: Preparation of
Figure imgf000100_0001
(S isomer at *)
2
g (3.82 mmol)
Figure imgf000100_0002
and HOBT 0.53 g (3.94 mmol) is dissolved in 5 mL DMF, diluted to 80 mL with CH2Cl2, and cooled to 0°C.
DCC, 0.81 g (3.79 mmol) is added, followed by a solution of CAD 0.93 g (3.82 mmol) in 25 mL
CH2Cl2/DMF. The mixture is stirred at room
temperature overnight and is evaporated under reduced pressure to an oil with suspended solids. The mixture is suspended in EtOAc, filtered, and
extracted with 1N citric acid, saturated NaCl
solution, saturated NaHCO3, and saturated NaCl solution. The organic phase is dried over MgSO4, filtered, and evaporated to an oil with suspended solids. The residue is suspended in Et2O, filtered, and evaporated under reduced pressure to a solid, 2.76 g. NMR spectroscopy confirms the structure.
Mass Spectroscopy further confirms the structure; MS (FAB) 712.2 (100.0%) M+. 2
Step C: Preparation of
Figure imgf000101_0004
OH
(S isomer at *) 2.55 g (3.58 mmol) (S isomer at *)
Figure imgf000101_0003
is dissolved in 70 mL 1 M t-butyl ammonium fluoride in THF. After stirring at 25°C for 6 hours, the mixture is evaporated under reduced pressure to an oil. The oil is suspended in 125 mL EtOAc and washed three times with 75 mL 1N HCl, three times with saturated NaHCO3, and saturated NaCl. The organic phase is dried over MgSO4, filtered, and evaporated to a solid 2.25 g. The solid is recrystallized from a mixture of CH2Cl2 and Et2O, and dried under reduced pressure to a solid, 0.75 g. NMR spectroscopy confirms the structure. Mass Spectroscopy further confirms the structure; MS (FAB) 598.2 (100.0%) M+. EXAMPLE S
(Asp•Boc•t-Bu)
Figure imgf000101_0001
Step A: Preparation of
(Asp•Boc•t-Bu)
Figure imgf000101_0002
N-Boc•Asp•O-t-Bu (17.1 g, 53 mmol), freed from the dicyclohexylamine salt by treatment with citric acid is dissolved in 300 mL of THF and
cooled to 0°C. CDI (8.6 g), the alcohol (11.09, 1.0 eq), and a catalytic
Figure imgf000102_0001
amount of DMAP are added and the mixture stirred at 0°C for 30 minutes and then refluxed for 24 hours. After cooling to room temperature, THF is removed by rotatory evaporation, and the residue dissolved in EtOAc and washed sequentially with 1N HCl, saturated solution of NaHCO3, and NaCl, and dried (magnesium sulfate). The crude product is purified by
chromatography with 10% to 20% EtOAc-hexane to afford 12.1 g of a white solid; MS (FAB) 367 (M-2 t-Bu+1) 324 (100%, M-Boc-t-Bu+1).
Step B: Preparation of
(Asp•Boc•t-Bu) -Phe-OBz
Figure imgf000102_0002
(Asp•Boc•t-Bu) 8.0 g
Figure imgf000102_0003
(16.7 mmol) is dissolved in 100 mL of EtOAc and a catalytic amount of 20% palladium on carbon is added and the mixture is stirred under one atmosphere of hydrogen for 2 hours. The mixture is filtered through celite, washed with EtOAc, and evaporated to dryness to afford an oil (~8.0 g). The crude product is dissolved in 70 mL of CH2Cl2, DCC (3.90 g,
1.1 eq), HOBT (2.5 g, 1.1 eq), DMAP (1.0 g, 0.5 eq), are added followed by Phe-OBz (from 5.0 g, 17.1 mmol of Phe-OBz•HCl). The mixture is stirred at room temperature overnight, DCU filtered, and the solution is washed with 1N HCl, saturated solution of NaHCO3, NaCl, and dried (magnesium sulfate). The product is purified by chromatography with 1% MeOH:CH2Cl2 to afford 11.4 g of a white solid; MS (FAB) 527
(M-Boc+1) 471 (M-Boc-t-Bu+1).
Step C: Preparation of
(Asp•Boc•t-Bu)
Figure imgf000103_0001
3
(Asp•Boc•t-Bu)O 4.56 g
Figure imgf000103_0002
3
(7.27 mmol) is dissolved in 80 mL of EtOAc and a catalytic amount of 20% palladium on carbon is added and the mixture stirred under one atmosphere of hydrogen for 4 hours. The mixture is filtered through celite, the solvent evaporated, and the product used without further purification.
EXAMPLE T
Atm(Cbz)-CAD
Step A: Boc-Atm(Cbz)-CAD
A mixture of Boc-Atm(Cbz) (European Published
Patent Application EP0399,556), 6.3 g (14.9 mmol), CAD, 4.0 g (1.1 eq), DCC, 3.4 g (1.1 eq), HOBT, 2.2 g (1.1 eq), in 50 mL of DMF is stirred at room
temperature overnight. The DCU is filtered and DMF removed under vacuum. The residue is taken up in 300 mL of EtOAc, washed with 1N citric acid,
saturated solution of NaHCO3, NaCl, and dried
(magnesium sulfate). The product is purified by chromatography with 1% MeOH:CH2Cl2 to afford 4.22 g of product as a white foam and a second portion of 7.5 g of product; MS (FAB) 647 (M+).
Step B: Atm(Cbz)-CAD
Boc-Atm(Cbz)-CAD, 2.5 g (3.86 mmol) is dissolved in 40 mL of CH2Cl2 and 3 mL of MeOH to afford a clear solution. Hydrogen chloride gas is passed through the solution for 10 minutes, and the reaction stirred for another 3.5 hours. The solvent is removed and the residue dissolved in 100 mL of EtOAc and 100 mL of NaHCO3. The layers are separated and the EtOAc layer is washed with NaCl and dried (magnesium sulfate). The solvent is evaporated to afford 2.2 g of product as a pale white foam which is used without further purification.
EXAMPLE U
(Asp•Boc•t-Bu) -Atm(Cbz)-CAD
(Asp•Boc•t-Bu)O 2 (7.27 mmol)
Figure imgf000104_0001
(Example S) is dissolved in 100 mL of DMF at 0°C and Atm(Cbz)-CAD (3.4 g, 6.1 mmol) (Example T), DCC
(1.51 g, 7.32 mmol), and HOBT (1.0 g, 7.31 mmol) are added. The mixture is stirred at room temperature overnight. The DCU is filtered, DMF is removed by rotatory evaporation, and the residue is dissolved in 200 mL of EtOAc, washed with 1N citric acid,
saturated solution of NaHCO3 and NaCl, and dried (magnesium sulfate). The product is purified by repeated chromatography (4X) with 1% MeOH:CH2Cl2 to afford 4.6 g of a yellowish foam; MS (FAB) 1066
(M+1)+. EXAMPLE V
Figure imgf000105_0001
Step A: Preparation of TBDMSO-CH
Figure imgf000105_0002
A mixture TBDMSO- - O2H (4.20 g, 18 mmol)
3
Figure imgf000105_0003
Phe-OBz (from 5.0 g, 17.1 mmol of Phe-OBz•HCl), DCC
(3.9 g, 1.1 eq), HOBT (2.5 g, 1.1 eq), DMAP (1.0 g,
0.5 eq) is dissolved in 200 mL of CH2Cl2. The
mixture is stirred at room temperature overnight, filtered, and the solution washed with 1N HCl, saturated NaHCO3 and NaCl, and dried (magnesium sulfate). The product is purified by chromatography with 15% EtOAc:hexane to afford 6.94 g of the product as a clear oil; MS (FAB) 470 (M+).
Step B: Preparation of TBDMSO-
Figure imgf000105_0004
TBDMSO-CH2- -OBz (2.0 g, 4.2 mmol) is
Figure imgf000105_0005
dissolved in 50 mL of EtOAc and a catalytic amount of 20% palladium on carbon is added and the mixture stirred under one atmosphere of hydrogen for 2 hours. The mixture is filtered, the solvent evaporated to afford 1.7 g of product which is used without further purification. EXAMPLE W
Figure imgf000106_0001
3
Step A: Preparation of TBDMSO- e-Atm(Cbz)-CAD
Figure imgf000106_0002
TBDMSO 2 1.7 g (Example V) and
Figure imgf000106_0003
Atm (Cbz)-CAD (1.0 eq) (Example T) are dissolved in 50 mL of DMF, cooled to 0°C, and DCC (960 mg, 1.2 eq) and HOBT (580 mg, 1.2 eq) added. The mixture is stirred at room temperature overnight, filtered, and DMF removed by rotatory evaporation. The residue is dissolved in 100 mL of EtOAc and washed with 1N citric acid, saturated NaHCO3 and NaCl, and dried (magnesium sulfate). The product is purified by chromatography with 1% to 2% MeOH:CH2Cl2 to afford 1.6 g of product as a white foam; MS (FAB) 908 (M+).
Step B: Preparation of 2 -Phe-Atm(Cbz)-CAD
Figure imgf000106_0004
TBDMSO- -Phe-Atm(Cbz)-CbD (340 mg,
Figure imgf000106_0005
0.37 mmol) is dissolved in 15 mL of MeOH and a catalytic amount of TsOH is added. The reaction mixture is heated to 60°C for 3 hours, MeOH
evaporated, and the residue taken up into water and EtOAc, and extracted with EtOAc (3 x 30 mL), and dried (magnesium sulfate). The product is purified by chromatography with 2% MeOH:CH2Cl2 to afford
280 mg of the product as a white foam; MS (FAB) 795 (M+1).
Step C: Preparation of
Figure imgf000107_0001
-Phe-Atm(Cbz)-CAD (1.1 g,
Figure imgf000107_0002
1.38 mmol) is dissolved in 20 mL of dry MeOH. TsOH
(530 mg, 0.2 eq) and a catalytic amount of 10% palladium on carbon are added and the mixture stirred under one atmosphere of hydrogen at room temperature for 24 hours. The mixture is filtered and the solvent evaporated. The product is purified by chromatography with 3% to 5% MeOH:CH2Cl2 to afford
700 mg of product as a light yellow foam; MS (CI) 660
(M+).
EXAMPLE X
3
2 -Phe-Alg-CAD
Figure imgf000107_0003
3
Step A: Preparation of TBDMSO- 2 -Alg-CAD
Figure imgf000107_0004
TBDMSO- 2 (1.02 g, 4.31 mmol),
Figure imgf000107_0005
Phe-Alg-CAD (1.00 g, 2.05 mmol) (Example N),, DCC
(907 mg, 4.40 mmol), HOBT (594 mg, 4.4 mmol), and
DMAP (268 mg, 220 mmol) are dissolved in 20 mL of CH2Cl2. The mixture is stirred at 0°C to room temperature for 40 hours, filtered, washed with 1N HCl, NaHCO3, NaCl, and dried (magnesium sulfate). The product is purified by chromatography with 1% to 2% MeOH:CH2Cl2 to afford 838 mg of the product as a white foam; MS (FAB) 702 (M+) 684 (M+-18).
Step B: Preparation of O 2 -Phe-Alg-CAD
Figure imgf000108_0001
3
TBDMSO-C -Phe-Alg-CAD (800 mg,
Figure imgf000108_0002
1.14 mmol) is dissolved in 15 mL of MeOH. A
catalytic amount of TsOH is added and the mixture heated to reflux for 2 hours. The MeOH is removed by rotatory evaporation. The product is purified by chromatography with 2% to 3% MeOH:CH2Cl2 to afford 523 mg of the product as a white foam; MS (FAB) 588 (M+) 570 (m+-18).

Claims

1. A compound of Formula I
A-E-G-J I wherein A is
wherein R is hydrogen or
Figure imgf000109_0001
alkyl of from one to six carbon atoms, wherein R is as defined above,
Figure imgf000109_0002
wherein R is as defined above,
Figure imgf000109_0003
Figure imgf000110_0001
Figure imgf000110_0002
wherein R1 is hydrogen, wherein R3 is hydrogen,
Figure imgf000110_0003
wherein n is zero or an integer of 1 or 2 and
Figure imgf000110_0004
R4 is hydrogen or
hydroxyl,
CH3-,
H2N-(CH2)4-,
HO2C-(CH2)m- wherein m is an integer of
1 or 2, or (CH2)m- wherein m is as defined
Figure imgf000110_0005
above, HO2CCH(OH)CH(OH)
Figure imgf000111_0002
O
O O O
Figure imgf000111_0001
X is O, S, or NH, and
R2 is alkyl of from one to six carbon atoms;
E is - wherein R5 is
Figure imgf000111_0003
Figure imgf000112_0001
wherein R6, R7, R8, or R9 are each
independently hydrogen, alkyl of from one to six carbon atoms, alkoxy of from one to six carbon atoms, halogen, or
trifluoromethyl,
wherein R1 and X are as defined above,
wherein R1 and X are as defined above,
Figure imgf000112_0002
wherein R1 is as defined above, 1 wherein X and R1 are as defined above, wherein R1 and X are as defined above, or wherein R1 and X are as defined above;
Figure imgf000113_0001
G is - wherein R5 is as defined above,
Figure imgf000113_0002
or
wherein R10 is
Figure imgf000113_0003
hydrogen,
alkyl of from one to six carbon atoms,
-CO2CH3,
Figure imgf000113_0004
-CH2-CH=CH2 ,
-CH2-C≡CH,
-CH2-CN,
-CH2-OH,
Figure imgf000114_0001
-CH2-CH2X-R1 wherein X and R1 are as defined above,
-CH2X-R1 wherein X and R1 are as defined above,
1 wherein X and R1 are as 3
Figure imgf000114_0002
defined above,
-CH2-CH2CH2CH2-NH2,
-CH2-CH2-S (O) n -R1 wherein n and R1 are as defined above,
- (CH2) n-CONH2 wherein n is as defined above,
Figure imgf000114_0003
wherein R1 is as defined above,
Figure imgf000114_0004
wherein X and R1 are as defined above; alternatively, E-G is
Figure imgf000115_0001
wherein R1, X, R5, and R10 are as defined above;
J is wherein R11 is
Figure imgf000115_0002
hydrogen,
alkyl,
Figure imgf000115_0003
R12 is
Figure imgf000115_0004
3
wherein R1 and X are as defined above, wherein R1 and X are as defined
Figure imgf000115_0005
above,
Figure imgf000115_0006
-CH2-OC2H5 and R1 and X are as defined above and p is zero or an integer of one, or wherein R14 is
Figure imgf000116_0001
or
Figure imgf000116_0002
-OC2H5 and R11 and p are as defined above;
provided R1 with the exclusion of R1 being hydrogen is encompassed within the definition of at least one of A, E, G, or J; or a
pharmaceutically acceptable salt thereof.
2. A compound according to Claim 1, in which A is
N wherein R is hydrogen or alkyl of from one to six carbon atoms,
Figure imgf000116_0003
Figure imgf000117_0001
Figure imgf000117_0002
wherein R1 is wherein R3 is
Figure imgf000117_0003
hydrogen,
Figure imgf000117_0004
CH3-,
H2N-(CH2)4-,
HO2C-(CH2)m- wherein m is an integer of 1 of 2, or wherein m is as defined
Figure imgf000117_0005
HO2C-CH(OH)CH(OH)
Figure imgf000117_0006
and
Figure imgf000118_0001
R2 is alkyl of from one to six carbon atoms ;
E is herein R5 is
wherein R6, R7, R8, or R9 are
Figure imgf000118_0002
each independently hydrogen, alkyl of from one to six carbon atoms, alkoxyl of from one to six carbon atoms , halogen or
trifluoromethyl,
Figure imgf000119_0001
wherein R1 is hydrogen, wherein R3 is
Figure imgf000119_0002
hydrogen,
Figure imgf000119_0003
CH3-,
H2N-(CH2)4-,
HO2C-(CH2)m- wherein m is an integer of
1 or 2, or
Figure imgf000119_0004
wherein m is as defined above, HO2C-CH(OH)CH(OH)
Figure imgf000119_0005
HO2C-CH2
Figure imgf000119_0006
Figure imgf000120_0001
G is wherein R5 is
Figure imgf000120_0002
wherein R6, R7, R8, or R9 are each independently hydrogen, alkyl of from one to six carbon atoms, alkoxy of from one to six carbon atoms, halogen, or trifluoromethyl,
Figure imgf000120_0003
wherein X is O, S, or NH and R1 is as defined above
1 wherein R1 and X are as
defined above,
wherein R1 is as defined above,
wherein X and R1 are as defined above,
wherein R1 and X are as defined above,
Figure imgf000121_0001
wherein R1 and X are as
Figure imgf000122_0001
defined above, or wherein R10 is
Figure imgf000122_0002
hydrogen,
alkyl of from one to six carbon atoms, -CO2CH3,
Figure imgf000122_0003
-CH2-CH=CH2,
-CH2-C≡CH,
-CH2-CN,
-CH2-OH,
- H -C H3 ,
Figure imgf000122_0004
-CH2-CH2X-R1 wherein X and R1 are as defined above,
- CH2X-R1 wherein X and R1 are as defined above,
wherein X and R1 are as defined
Figure imgf000122_0005
above,
-CH2-CH2CH2CH2-NH2,
-CH2-CH2-S(O)n-R1 wherein n is zero or an integer of 1 or 2 and R1 is as defined above,
- (CH2)n-CONH2 wherein n is as defined above,
Figure imgf000123_0001
wherein R1 is as defined above,
Figure imgf000123_0002
wherein X and R1 are as defined above;
alternatively, E-G is
Figure imgf000123_0003
R
wherein R1, X, R5, and R10 are as defined above;
J is 12 wherein R11 is
Figure imgf000123_0004
hydrogen,
alkyl,
, or
Figure imgf000123_0005
Figure imgf000124_0001
R12 is wherein R 1 is as defined above,
Figure imgf000124_0002
wherein R1 is as defined
above,
Figure imgf000124_0003
OR or
Figure imgf000124_0004
-CH2-OC2H5 and R1 and X are as defined above or 2 wherein R14 is
2
or
Figure imgf000124_0005
-OC2H5 and R11 is as defined above; provided R1 with the exclusion of R1 being hydrogen is encompassed within the definition of at least one of A, E, G, or J.
3. A compound according to Claim 2, in which J is
wherein R11 is
Figure imgf000124_0006
alkyl, , or R12 is 2 3
or
Figure imgf000125_0001
OR
and
Figure imgf000125_0002
R1 is hydrogen, wherein R3 is
Figure imgf000125_0003
hydrogen,
Figure imgf000125_0004
CH3-,
H2N-(CH2)4-,
HO2C-(CH2)m- wherein m is an integer of 1 or 2, or
Figure imgf000125_0005
wherein m is as defined above,
O
HO2C-CH(OH)CH(OH)
Figure imgf000126_0001
and X is 0,
Figure imgf000126_0002
H S, or NH, or wherein R14 is
, or
Figure imgf000126_0003
-OC2H5 and
R11 is as defined above.
4. A compound according to Claim 3 selected from the group consisting of:
SMO-Phe-Ser(Phe)-CAD;
SMO-Phe-Ser(Gln)-CAD; SMO-Phe-Ser(Pro)-CAD;
SMO-Phe-Ser(Glu)-CAD;
SMO-Phe-Ser(Lys)-CAD;
SMO-Phe-Ser(Asp)-CAD;
SMO-Phe-Ser(Gly)-CAD;
SMO-Phe-Ser(Ala)-CAD;
SMO-Phe-Ser(COCH2CH2CO2H)-CAD;
SMO-Phe-Ser(P(O) (OH)2)-CAD;
SMO-Phe-Ser(COCH(OH)CH(OH)CO2H)-CAD;
SMO-Phe-Thr(Phe)-CAD;
SMO-Phe-Thr(Gln)-CAD;
SMO-Phe-Thr(Pro)-CAD;
SMO-Phe-Thr(Glu)-CAD;
SMO-Phe-Thr(Lys)-CAD;
SMO-Phe-Thr(Asp)-CAD;
SMO-Phe-Thr(Gly)-CAD;
SMO-Phe-Thr(Ala)-CAD;
SMO-Phe-Thr(COCH2CH2CO2H)-CAD;
SMO-Phe-Thr(P(O) (OH)2)-CAD;
SMO-Phe-Thr(COCH(OH)CH(OH)CO2H)-CAD;
Boc-Tyr(Phe)-Pgy-CAD;
Boc-Tyr(Gln)-Pgy-CAD;
Boc-Tyr(Pro)-Pgy-CAD;
Boc-Tyr(Glu)-Pgy-CAD;
Boc-Tyr(Lys)-Pgy-CAD;
Boc-Tyr(Asp)-Pgy-CAD;
Boc-Tyr(Gly)-Pgy-CAD;
Boc-Tyr(Ala)-Pgy-CAD;
Boc-Tyr(COCH2CH2CO2H)-Pgy-CAD;
Boc-Tyr(P(O)(OH)CH(OH)CO2H)-Pgy-CAD;
Boc-Tyr(COCH(OH)CH(OH)CO2H)-Pgy-CAD;
SMO-Phe-Hse(Phe)-CAD;
SMO-Phe-Hse(Gln)-CAD;
SMO-Phe-Hse(Pro)-CAD;
SMO-Phe-Hse(Glu)-CAD;
SMO-Phe-Hse(Lys)-CAD; SMO-Phe-Hse(Asp)-CAD;
SMO-Phe-Hse(Gly)-CAD;
SMO-Phe-Hse(Ala)-CAD;
SMO-Phe-Hse(COCH2CH2CO2H)-CAD;
SMO-Phe•Hse(P(O)(OH)2)-CAD;
SMO-Phe Hse(COCH(OH)CH(OH)CO2H)-CAD;
SMO-Phe-Mal-CAD(2•Phe );
SMO-Phe-Mal-CAD(2•Gln);
SMO-Phe-Mal-CAD(2•Pro);
SMO-Phe-Mal-CAD(2•Glu);
SMO-Phe-Mal-CAD(2•Lys);
SMO-Phe-Mal-CAD(2•Asp);
SMO-Phe-Mal-CAD(2•Gly);
SMO-Phe-Mal-CAD(2•Ala);
SMO-Phe-Mal-CAD(2•CCOOCCH2CH2CO2H);
SMO-Phe-Mal-CAD(2•PP((CO)(OH)2);
SMO-Phe-Mal-CAD(2•COCH(OH)CH(OH)CO2H);
SMO-Phe-Atm-CAD(2•Phe);
SMO-Phe-Atm-CAD(2•Gln);
SMO-Phe-Atm-CAD(2•Pro);
SMO-Phe-Atm-CAD(2•Glu);
SMO-Phe-Atm-CAD(2•Lys);
SMO-Phe-Atm-CAD(2•Asp);
SMO-Phe-Atm-CAD(2•Gly);
SMO-Phe-Atm-CAD[2•Ala);
SMO-Phe-Atm-CAD,2•COCH2CH2CO2H);
SMO-Phe-Atm-CAD(2•P(O)(OH)2);
SMO-Phe-Atm-CAD (2•COCH(OH)CH(OH)CO2H);
SMO-Phe-Atm(Phe)-CAD;
SMO-Phe-Atm(Gln)-CAD;
SMO-Phe-Atm(Pro)-CAD;
SMO-Phe-Atm(Glu)-CAD;
SMO-Phe-Atm(Lys)-CAD;
SMO-Phe-Atm(Asp)-CAD;
SMO-Phe-Atm(Gly)-CAD;
SMO-Phe-Atm(Ala)-CAD; SMO-Phe-Atm(COCH2CH2CO2H)-CAD;
SMO-Phe-Atm(P(O)(OH)2)-CAD;
SMO-Phe-Atm(COCH(OH)CH(OH)CO2H)-CAD;
(Phe)O-CH2 -Phe-Alg-CAD; (Gln)O-CH2 -Phe-Alg-CAD;
(Pro)O-CH2 -Phe-Alg-CAD;
(Glu)O-CH2 -Phe-Alg-CAD;
(Lys)O-CH2 -Phe-Alg-CAD;
(Asp)O-CH2 -Phe-Alg-CAD; (Gly)O-CH2 -Phe-Alg-CAD;
(Ala)O-CH2 -Phe-Alg-CAD;
Figure imgf000129_0001
(HO2CCH2CH2CO)O-CH2-
Figure imgf000129_0002
-Phe-Alg-CAD;
((HO)2P(O))OCH2 -Phe-Alg-CAD;
Figure imgf000129_0003
(HO2CCH(OH)CH(OH)CO)OCH2
Figure imgf000129_0004
-Phe-Alg-CAD;
(Phe)O-CH2-C(CH3)2 -Phe-Alg-CAD;
(Gln)O-CH2-C(CH3)2 -Phe-Alg-CAD;
(Pro)O-CH2-C(CH3)2 -Phe-Alg-CAD;
(Glu)O-CH2-C(CH3)2
Figure imgf000129_0005
-Phe-Alg-CAD; (Lys)O-CH2-C(CH3)2 -Phe-Alg-CAD;
(Asp)O-CH2-C(CH3)2 -Phe-Alg-CAD;
(Gly)O-CH2-C(CH3)2 -Phe-Alg-CAD;
(Ala)O-CH2-C(CH3)2 -Phe-Alg-CAD;
Figure imgf000130_0001
(HO2CCH2CH2CO)O-CH2-C(CH3)2
Figure imgf000130_0002
Phe-Alg-CAD;
((HO)2P(O))O-CH2-C(CH3)2-
Figure imgf000130_0003
-Phe-Alg-CAD;
(HO2CCH(OH)CH(OH)CO)O-CH2-C(CH3)2
Figure imgf000130_0004
-Phe-Alg-CAD;
(Phe)O-CH2- Phe-Atm-CAD;
(Gln)O-CH2 Phe-Atm-CAD;
(Pro)O-CH2 Phe-Atm-CAD;
(Glu)O-CH2 Phe-Atm-CAD;
(Lys)O-CH2 Phe-Atm-CAD;
(Asp)O-CH2 -Phe-Atm-CAD;
(Gly)O-CH2 -Phe-Atm-CAD;
(Ala)O-CH2 -Phe-Atm-CAD;
Figure imgf000130_0005
(HO2CCH2CH2CO)O-CH2- -Phe-Atm-CAD;
Figure imgf000130_0006
((HO)2P(O)O-CH2- -Phe-Atm-CAD;
(HO2CCH(OH)CH(OH)CO)O-CH2-
Figure imgf000131_0001
-Phe-Atm-CAD;
0
(Phe)O-CH2-C(CH3)2- -Phe-Atm-CAD;
(Gln)O-CH2-C(CH3)2- -Phe-Atm-CAD;
(Pro)O-CH2-C(CH3)2- -Phe-Atm-CAD;
(Glu)O-CH2-C(CH3)2- -Phe-Atm-CAD;
(Lys)O-CH2-C(CH3)2- -Phe-Atm-CAD;
(Asp)O-CH2-C(CH3)2- -Phe-Atm-CAD; (Gly)O-CH2-C(CH3)2- -Phe-Atm-CAD;
(Ala)O-CH2-C(CH3)2-
Figure imgf000131_0002
-Phe-Atm-CAD; (HO2CCH2CH2CO)O-CH2-C(CH3)2-
Figure imgf000131_0003
-Phe-Atm-CAD;
((HO)2P(O))O-CH2-C(CH3)2
Figure imgf000131_0004
-Phe-Atm-CAD;
(HO2CCH(OH)CH(OH)CO)O-CH2-C(CH3)2- -Phe-Atm-CAD;
Figure imgf000131_0005
2
Figure imgf000131_0006
Figure imgf000132_0001
5. A method of treating renin-associated
hypertension comprising administering to a host suffering therefrom a therapeutically effect amount of a compound according to Claim 1 in unit dosage form.
6. A pharmaceutical composition adapted for
administration as an antihypertensive agent comprising a therapeutically effective amount of a compound according to Claim 1 in admixture with a pharmaceutically acceptable excipient, diluent, or carrier.
7. A method of treating hyperaldosteronism
comprising administering to a host suffering therefrom a therapeutically effective amount of a compound according to Claim 1 in unit dosage form.
8. A pharmaceutical composition adapted for
administration as an agent for treating
hyperaldosteronism comprising a therapeutically effective amount of a compound according to Claim 1 in admixture with a pharmaceutically acceptable excipient, diluent, or carrier.
9. A method of treating congestive heart failure comprising administering to a host suffering therefrom a therapeutically effective amount of a compound according to Claim 1 in unit dosage form.
10. A pharmaceutical, composition adapted for
administration as an agent for treating
congestive heart failure comprising a
therapeutically effective amount of a compound according to Claim 1 in admixture with a pharmaceutically acceptable excipient, diluent, or carrier.
11. A method of treating glaucoma comprising
administering to a host suffering therefrom a. therapeutically effective amount of a compound according to Claim 1 in unit dosage form.
12. A pharmaceutical composition adapted for
administration as an agent for treating glaucoma comprising a therapeutically effective amount of a compound according to Claim 1 in admixture with a pharmaceutically acceptable excipient, diluent, or carrier.
13. A method of preparing a compound having the
Formula I
A-E-G-J I wherein A is
wherein R is hydrogen or alkyl of from one to six carbon atoms,
wherein R is as defined above,
Figure imgf000134_0001
\
wherein R is as defined above,
0
0
Figure imgf000135_0001
( or
Figure imgf000135_0002
wherein R1 is hydrogen,
Figure imgf000135_0003
wherein R3 is
Figure imgf000135_0004
hydrogen, wherein n is zero or an integer of 1 or 2 and R4 is hydrogen or
Figure imgf000136_0001
hydroxyl,
CH3-,
H2N-(CH2)4-,
HO2C-(CH2)m- wherein m is an integer of
1 or 2, or
H 2 - wherein m is as defined
Figure imgf000136_0002
above,
HO2CCH (OH) CH (OH)
Figure imgf000136_0003
O2 2 2 2 O O O
Figure imgf000136_0004
X is O, S, or NH, and
R2 is alkyl of from one to six carbon atoms;
E is wherein R5 is
Figure imgf000137_0001
wherein R6, R7, R8, or R9 are each independently hydrogen, alkyl of from one to six carbon atoms,
Figure imgf000137_0002
alkoxy of from one to six carbon atoms, halogen, or trifluoromethyl,
1
wherein R1 and X are as defined above,
wherein R1 and X are as defined above,
Figure imgf000137_0003
wherein R1 is as defined above,
wherein X and R1 are as defined above,
wherein R1 and X are as defined above, or
wherein R1 and X are as defined above;
Figure imgf000138_0001
G is - wherein R5 is as defined above,
Figure imgf000138_0002
or
wherein R10 is
Figure imgf000138_0003
hydrogen,
alkyl of from one to six carbon atoms,
-CO2CH3,
Figure imgf000139_0001
-CH2-CH=CH2,
-CH2-C≡CH,
-CH2-CN,
-CH2-OH,
- - 3 -
Figure imgf000139_0002
-CH2-CH2X-R1 wherein X and R1 are as defined above,
-CH2X-R1 wherein X and R1 are as defined
above,
wherein X and R1 are as defined
Figure imgf000139_0003
above,
- CH2-CH2CH2CH2-NH2,
-CH2-CH2-S(O)n-R1 wherein n and R1 are as defined above,
-(CH2)n-CONH2 wherein n is as defined above,
wherein R1 is as defined above, , or
Figure imgf000139_0005
Figure imgf000140_0001
wherein X and R1 are as defined above;
alternatively, E-G is
Figure imgf000140_0002
wherein R1, X, R5, and R10 are as defined above;
J is wherein R11 is
Figure imgf000140_0003
hydrogen,
alkyl,
or
Figure imgf000140_0004
R12 is
Figure imgf000140_0005
3
wherein R1 and X are as defined above, wherein R1 and X are as defined
Figure imgf000140_0006
above, or
Figure imgf000140_0007
-CH2-OC2H5 and R1 and X are as defined above and p is zero or an integer of one, or wherein R14 is
Figure imgf000141_0001
- or
Figure imgf000141_0002
-OC2H5 and R11 and p are as defined above;
provided R1 with the exclusion of R1 being hydrogen is encompassed within the definition of at least one of A, E, G, or J; or a
pharmaceutically acceptable salt thereof
comprises:
a) coupling a compound of Formula II
A'-E'-G'-J' II wherein A', E', G', and J' are as defined above for A, E, G, and J provided R1 is hydrogen and is encompassed within the definition of at least one of A', E', G', or J' with a compound of Formula III
R1a-XH
III wherein R1a is as defined above for R1 but excluding R1 is hydrogen and providing any basic or acidic groups contain conventional protecting groups and X is as defined above to afford a compound of Formula IV A''-E'' -G" -J"
IV wherein A'', E'', G" , and J'' are as defined above for A, E, G, and J provided R1a is as defined above and is encompassed within the definition of at least one of A", E'', G" or
J";
b) a compound of Formula IV is
deprotected in a conventional manner to afford a compound of Formula I; and if desired,
converting a compound of Formula I to a
corresponding pharmaceutically acceptable salt by conventional means and, if so desired, converting the corresponding pharmaceutically acceptable salt to a compound of Formula I by conventional means.
PCT/US1992/007463 1991-09-17 1992-09-01 Novel amino acid prodrug renin inhibitors WO1993006127A1 (en)

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