Anti-hypertensive molecules and process for preparation thereof

Abstract

The present invention relates to novel anti-hypertensive molecules. The present invention also provides a process for the preparation of novel antihypertensive molecules. The present invention particularly relates to the preparation of novel Angiotensin Converting Enzyme Inhibitors (ACEI) with prolonged activity. ACE inhibitors play an important role in Re -Angiotensin-Aldosteron system (RAAS) by inhibiting the activity of Angiotensin Converting Enzyme (ACE) and therefore are used to regulate blood pressure. ACE inhibitors synthesized by the process of present invention have a peptide moiety and nonpeptide moiety. ACE inhibitors, synthesized by this present invention, show enhanced bioavailability and fewer side effects.

1 . A peptide derivative peptidomimic having general formula X-CX 1 -NH-AA 1 -CONH-AA 2  wherein X is a heterocyclic or unusual amino acid, X 1  is O or H 2  and AA 1  and AA 2  are amino acids. 
     
     
         2 . A peptide derivative according to  claim 1 , wherein X is a heterocyclic selected from the group consisting of F-moc-3-(2-furyl)-L-alanine, F-3-(3-thienyl)-L-alanine, 4-Fmoc-piperazine-1-yl-acetic acid hydrate, Fmoc-3,3-diphenyl-L-alanine, 1-Fmoc-azetidine-3-carboxylic acid, Benzimidazolepropionic acid, Fmoc1, 2,3,4 tetrahdroquinoline-3-carboxylic acid, 2-oxo-4-phenyl-3-oxazolidine-acetic acid, 5-Methoxy-2-methyl-3-indole acetic acid and 5-Mercapto-1-terazole acetic acid. 
     
     
         3 . A peptide derivative according to  claim 1 , wherein X is an unusual amino acids selected from a group consisting of 5-Hydroxytryptophan, L-Abrine, L-β-homoproline, β-HomoTrp -OH, Homophenylalanine L-β-homotryptophan, L-2-propargyl glycine,  3 , 3  Diphenylalanine, L-β-Homohydroxyproline and Cyclohexylalanine. 
     
     
         4 . A peptide derivative according to  claim 1 , wherein the dipeptide for position AA 1 -AA 2 , is selected from the group consisting of Orn-Pro, Cha-Pro, Ile-Pro, Dap-Pro, Val-Tip, Lys-Pro, Lys-Trp, Orn-Trp, Dap-Trp, Ile-Phe, β-Ala-Pro, Pro-Pro and Cha-Trp. 
     
     
         5 . A peptide derivative according to  claim 1 , wherein the derivative represented by general formula X-CX1-NH-AA 1 -CONH-AA 2  is selected from the group consisting of:
 (a) L-Abrine-Orn-Pro, 3-(3-thienyl)-L-alanine-Orn-Pro, 3-(2-furyl)-L-alanine-Orn-Pro, 2-Benzimidazoleacetic acid-Orn-Pro, 5-Hydroxytrytophan-Orn-Pro, Homotryptophan-Orn-Pro, Homophenyalanine-Orn-Pro, 1,2,3,4-tetrahydro isoquinoline-3-carboxylic acid-Orn-Pro, Azetidine-3-carboxylic acid-Orn-Pro, Cyclohexylalanine-Orn-Pro, 2-Oxo-4-phenyl-3-oxazolidine acetic acid-Orn-Pro, 4-piperazine acetic acid-Orn-Pro   (b) L-Abrine-Cha-Pro, 3-(3-thienyl)-L-alanine-Cha-Pro, 3-(2-furyl)-L-alanine-Cha-Pro, 2-Benzimidazoleacetic acid-Cha-Pro, 5-Hydroxytrytophan-Cha-Pro, Homotryptophan-Cha-Pro, Homophenyalanine-Cha-Pro, 1,2,3,4-tetrahydro isoquinoline-3-carboxylic acid-Cha-Pro, Azetidine-3-carboxylic acid-Cha-Pro, Cyclohexylalanine-Cha-Pro, 2-Oxo-4-phenyl-3-oxazolidine acetic acid-Cha-Pro, 4-piperazine acetic acid-Cha-Pro   (c) L-Abrine-Ile-Pro, 3-(3-thienyl)-L-alanine-Ile-Pro, 3-(2-furyl)-L-alanine-Ile-Pro, 2-Benzimidazoleacetic acid-Ile-Pro, 5-Hydroxytrytophan-Ile-Pro, Homotryptophan-Ile-Pro Homophenyalanine-Ile-Pro, 1,2,3,4-tetrahydro isoquinoline-3-carboxylic acid-Ile-Pro, Azetidine-3-carboxylic acid-Ile-Pro, Cyclohexylalanine-Ile-Pro, 2-Oxo-4-phenyl-3-oxazolidine acetic acid-Ile-Pro, 4-piperazine acetic acid-Ile-Pro   (d) L-Abrine-Dap-Pro, 3-(3-thienyl)-L-alanine-Dap-Pro, 3-(2-furyl)-L-alanine-Dap-Pro, 2-Benzimidazoleacetic acid-Dap-Pro, 5-Hydroxytrytophan-Dap-Pro, Homotryptophan-Dap-Pro, Homophenyalanine-Dap-Pro, 1,2,3,4-tetrahydro isoquinoline-3-carboxylic acid-Dap-Pro, Azetidine-3-carboxylic acid-Dap-Pro, Cyclohexylalanine-Dap-Pro, 2-Oxo-4-phenyl-3-oxazolidine acetic acid-Dap-Pro, 4-piperazine acetic acid-Dap-Pro.   (e) L-Abrine-Val-Trp, 3-(3-thienyl)-L-alanine-Val-Trp, 3-(2-furyl)-L-alanine-Val-Trp, 2-Benzimidazoleacetic acid-Val-Trp, 5-Hydroxytrytophan-Val-Trp, Homotryptophan-Val-Trp, Homophenyalanine-Val-Trp,  1  , 2 , 3 , 4 -tetrahydro isoquinoline-3-carboxylic acid-Val-Trp, Azetidine-3-carboxylic acid-Val-Trp, Cyclohexylalanine-Val-Trp, 2-Oxo-4-phenyl-3-oxazolidine acetic acid-Val-Trp, 4-piperazine acetic acid-Val-Trp   (f) L-Abrine-Lys-Pro, 3-(3-thienyl)-L-alanine-Lys-Pro, 3-(2-furyl)-L-alanine-Lys-Pro, 2-Benzimidazoleacetic acid-Lys-Pro, 5-Hydroxytrytophan-Lys-Pro, Homotryptophan-Lys-Pro, Homophenyalanine-Lys-Pro, 1,2,3,4-tetrahydro isoquinoline-3-carboxylic acid-Lys-Pro, Azetidine-3-carboxylic acid-Lys-Pro, Cyclohexylalanine-Lys-Pro, 2-Oxo-4-phenyl-3-oxazolidine acetic acid-Lys-Pro, 4-piperazine acetic acid-Lys-Pro.   (g) L-Abrine-Lys-Trp, 3-(3-thienyl)-L-alanine-Lys-Trp, 3-(2-furyl)-L-alanine-Lys-Trp, 2-Benzimidazoleacetic acid-Lys-Trp, 5-Hydroxytrytophan-Lys-Trp, Homotryptophan-Lys-Trp, Homophenyalanine-Lys-Trp, 1,2,3,4-tetrahydro isoquinoline-3-carboxylic acid-Lys-Trp, Azetidine-3-carboxylic acid-Lys-Trp, Cyclohexylalanine-Lys-Trp, 2-Oxo-4-phenyl-3-oxazolidine acetic acid-Lys-Trp, 4-piperazine acetic acid-Lys-Trp   (h) L-Abrine-Orn-Trp, 3-(3-thienyl)-L-alanine-Orn-Trp, 3-(2-furyl)-L-alanine-Orn-Trp, 2-Benzimidazoleacetic acid-Orn-Trp, 5-Hydroxytrytophan-Orn-Trp, Homotryptophan-Orn-Trp, Homophenyalanine-Orn-Trp, 1,2,3,4-tetrahydro isoquinoline-3-carboxylic acid-Orn-Trp, Azetidine-3-carboxylic acid-Orn-Trp, Cyclohexylalanine-Orn-Tip, 2-Oxo-4-phenyl-3-oxazolidine acetic acid-Orn-Trp, 4-piperazine acetic acid-Orn-Trp.   (i) L-Abrine-Dap-Trp, 3-(3-thienyl)-L-alanine-Dap-Trp, 3-(2-furyl)-L-alanine-Dap-Trp, 2-Benzimidazoleacetic acid-Dap-Trp, 5-Hydroxytrytophan-Dap-Trp, Homotryptophan-Dap-Trp, Homophenyalanine-Dap-Trp, 1,2,3,4-tetrahydro isoquinoline-3-carboxylic acid-Dap-Trp, Azetidine-3-carboxylic acid-Dap-Trp, Cyclohexylalanine-Dap-Trp, 2-Oxo-4-phenyl-3-oxazolidine acetic acid-Dap-Trp, 4-piperazine acetic acid-Dap-Trp.   (j) L-Abrine-Ile-Phe, 3-(3-thienyl)-L-alanine-Ile-Phe, 3-(2-furyl)-L-alanine-Ile-Phe, 2-Benzimidazoleacetic acid-Ile-Phe, 5-Hydroxytrytophan-Ile-Phe, Homotryptophan-Ile-Phe, Homophenyalanine-Ile-Phe, 1,2,3,4-tetrahydro isoquinoline-3-carboxylic acid-Ile-Phe, Azetidine-3-carboxylic acid-Ile-Phe, Cyclohexylalanine-Ile-Phe, 2-Oxo-4-phenyl-3-oxazolidine acetic acid-Ile-Phe, 4-piperazine acetic acid-Ile-Phe.   (k) L-Abrine-β-Ala-Pro, 3-(3-thienyl)-L-alanine-β-Ala-Pro, 3-(2-furyl)-L-alanine-β-Ala-Pro, 2-Benzimidazoleacetic acid-β-Ala-Pro, 5-Hydroxytrytophan-β-Ala-Pro, Homotryptophan-p-Ala-Pro, Homophenyalanine-β-Ala-Pro, 1,2,3,4-tetrahydro isoquinoline-3-carboxylic acid-β-Ala-Pro, Azetidine-3-carboxylic acid-β-Ala-Pro, Cyclohexylalanine-β-Ala-Pro, 2-Oxo-4-phenyl-3-oxazolidine acetic acid-β-Ala-Pro, 4-piperazine acetic acid-β-Ala-Pro.   (l) L-Abrine-Pro-Pro, 3-(3-thienyl)-L-alanine-Pro-Pro, 3-(2-furyl)-L-alanine-Pro-Pro, 2-Benzimidazoleacetic acid-Pro-Pro, 5-Hydroxytrytophan-Pro-Pro, Homotryptophan-Pro-Pro, Homophenyalanine-Pro-Pro, 1,2,3,4-tetrahydro isoquinoline-3-carboxylic acid-Pro-Pro, Azetidine-3-carboxylic acid-Pro-Pro, Cyclohexylalanine-Pro-Pro, 2-Oxo-4-phenyl-3-oxazolidine acetic acid-Pro-Pro, 4-piperazine acetic acid-Pro-Pro.   (m) L-Abrine-Cha-Trp, 3-(3-thienyl)-L-alanine-Cha-Trp, 3-(2-furyl)-L-alanine-Cha-Trp, 2-Benzimidazoleacetic acid-Cha-Trp, 5-Hydroxytrytophan-Cha-Trp, Homotryptophan-Cha-Trp, Homophenyalanine-Cha-Trp, 1,2,3,4-tetrahydro isoquinoline-3-carboxylic acid-Cha-Trp, Azetidine-3-carboxylic acid-Cha-Trp, Cyclohexylalanine-Cha-Trp, 2-Oxo-4-phenyl-3-oxazolidine acetic acid-Cha-Trp, 4-piperazine acetic acid-Cha-Trp.   
     
     
         6 . (canceled) 
     
     
         7 . A peptidomimic compound according to  claim 1 , wherein the concentration of the peptidomimic compound for 50% inhibition of ACE activity (IC 50 ) ranged from 2 μmole to 10 μmole in in-vitro condition using synthetic substrate Hippuryl-Histidyl-Leucine (HHL). 
     
     
         8 . A peptidomimic compound according to  claim 1 , wherein the dose of the synthesized ACE inhibiting peptidomimic compound which effectively blocked angiotensin converting enzyme ranges between 5-8 mg/kg of body weight. 
     
     
         9 . A process to synthesize peptide derivative peptidomimics comprising
 (a) coupling ACE inhibiting antihypertensive peptidomimic molecule wherein a heterocyclic or unusual amino acid present at ante-penultimate position is coupled to a dipeptide with amino acids present at ultimate position and penultimate position;   (b) synthesizing dipeptide on a solid support by coupling and deprotection;   (c) coupling the heterocyclic or unusual amino acid to deprotected dipeptide at the -terminal of dipeptide;   (d) cleaving the synthesized peptidomimic compound of step (c) from solid support followed by purification and characterization;   
     
     
         10 . A process according to  claim 9 , wherein the solid support used is selected from polystyrene resins linked with a suitable agent/handles. 
     
     
         11 . A process according to  claim 10 , wherein the agent is acid labile and comprises 4-hydroxymethylphenoxyacetic acid. 
     
     
         12 . A process according to  claim 10 , wherein the agent is hyper acid labile and is selected from the group consisting of 4-hydroxymethyl-3-methoxy-phenoxyacetic acid and 2-chlorotrityl-2-chloride linker. 
     
     
         13 . A process according to  claim 9 , wherein the anchoring of activated C-terminal of the—-protected amino acid on to the solid support is carried out by symmetrical anhydrides. 
     
     
         14 . A process according to  claim 9 , wherein the anchoring of activated C-terminal of the—-protected amino acid on to the solid support is carried out by reactive ester formation using 1-hydroxybenzotriazole, benzotriazolyloxy-tridimethylamino-phosphonium-hexaflourophosphate and benzotriazole-1-yl-oxy-trispyrrolidino-phosphoniunm-hexaflourophosphate. 
     
     
         15 . A process as claimed in  claim 9 , wherein deprotection of the—α-protected amino acid is carried out removing flouro-methyl-oxy carbonyl group depending on the compatibility with the linking group on the solid support. 
     
     
         16 . A process according to  claim 9 , wherein the cleavage is effected using trofluroacetic acid, acetic acid or trifluroethanol depending upon the linking agent/functional group attached on the solid support and the C terminal functional group desired. 
     
     
         17 . A process according to  claim 9 , wherein the purification is carried out by gel permeation method using Sephadex G/LH-20 followed by characterization using techniques of HPLC, MALDI-T of and LC-MS. 
     
     
         18 . (canceled) 
     
     
         19 . (canceled) 
     
     
         20 . Method for the inhibition of angiotensin converting enzyme in a subject suffering from hypertension comprising administering a pharmaceutically effective amount of a peptide derivative peptidomimic having general formula X-CX1-NH-AA 1 -CONH-AA 2  wherein X is a heterocyclic or unusual amino acid, X 1  is O or H 2  and AA 1  and AA 2  are amino acids to the subject with a pharmaceutically effective carrier. 
     
     
         21 . Method according to  claim 20 , wherein the subject is a mammal. 
     
     
         22 . Method according to  claim 20 , wherein the subject is a human being. 
     
     
         23 . Method according to  claim 20 , wherein dose of the synthesized ACE inhibiting peptidomimic compound which effectively blocked angiotensin converting enzyme ranges between 5-8 mg/kg of body weight.