WO2001068114A1 - Nouveaux peptides a activite hypotensive - Google Patents

Nouveaux peptides a activite hypotensive Download PDF

Info

Publication number
WO2001068114A1
WO2001068114A1 PCT/US2001/007530 US0107530W WO0168114A1 WO 2001068114 A1 WO2001068114 A1 WO 2001068114A1 US 0107530 W US0107530 W US 0107530W WO 0168114 A1 WO0168114 A1 WO 0168114A1
Authority
WO
WIPO (PCT)
Prior art keywords
pro
lys
asp
glu
ala
Prior art date
Application number
PCT/US2001/007530
Other languages
English (en)
Inventor
Foe Siong Tjoeng
Zhonghong Guan
Wei Li
Kathy Mandrell
Min Liu
Xiangra Chen
Original Assignee
Monsanto Company
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Monsanto Company filed Critical Monsanto Company
Priority to AU2001247334A priority Critical patent/AU2001247334A1/en
Publication of WO2001068114A1 publication Critical patent/WO2001068114A1/fr

Links

Classifications

    • 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
    • C07K5/08—Tripeptides
    • C07K5/0821—Tripeptides with the first amino acid being heterocyclic, e.g. His, Pro, Trp
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/152—Milk preparations; Milk powder or milk powder preparations containing additives
    • A23C9/1526—Amino acids; Peptides; Protein hydrolysates; Nucleic acids; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/17—Amino acids, peptides or proteins
    • A23L33/18—Peptides; Protein hydrolysates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/06—Tripeptides
    • 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
    • C07K5/08—Tripeptides
    • C07K5/0802—Tripeptides with the first amino acid being neutral
    • C07K5/0812—Tripeptides 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
    • C07K5/08—Tripeptides
    • C07K5/0819—Tripeptides with the first amino acid being acidic
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs

Definitions

  • This invention relates to novel tripeptides which exhibit anti-hypertensive properties. These novel anti- hypertensive tripeptides may be used as active ingredients in food products, pharmaceutical compositions, and dietary supplements.
  • the present invention also relates to methods of treatment and prophylaxis of hypertension, myocardial infarction, left ventricular systolic dysfunction, diabetes mellitus, progressive renal failure and congestive heart failure using such novel tripeptides.
  • Hypertension is generally clinically defined as a systolic blood pressure greater than 140 mmHg or a diastolic blood pressure greater than 90 mmHg. Hypertension is the primary risk factor for coronary, cerebral, and renal vascular diseases which cause over half of all deaths in the United States. The widespread awareness of the danger of elevated blood pressure has become the most frequent reason for visits to physicians.
  • primary hypertension No single or specific cause is known for the hypertension referred to as primary (essential) hypertension.
  • Primary hypertension has been attributed to such causes as hemodynamic pattern, genetic predisposition, vascular hypertrophy, hyperinsulinemia, defects in cell transport of binding, defects in the reninangiotensin system (low-renin or high renin hypertension) and along with insulin, angiotensin and natriuretic hormone, catecholamines arising in response to stress are known to be pressor-growth promoters.
  • Increased sympathetic nervous activity may raise the blood pressure in a number of ways, for example, either alone or in concert with stimulation of renin release by catecholamines, causing arteriolar and venous constriction, by increasing cardiac output, or by altering the normal renal pressure-volume relationship. Also, recent evidence demonstrates that sodium has a causal role in the genesis of hypertension. Primary hypertension is also associated with, for example, obesity, sleep apnea, physical inactivity, alcohol intake, smoking, diabetes mellitus, polycythemia and gout.
  • hypertension Secondary forms of hypertension may arise from oral contraceptive use and parenchymal renal disease: renovascular hypertension caused by, for example, atherosclerotic disease, tumors (renin-secretory tumors) ; Cushing's Syndrome; heart surgery; and pregnancy. Chronic hypertension and renal disease during pregnancy may progress into eclampsia, a primary cause of fetal death.
  • Essential hypertension is a relatively common disease state in humans . This disease has been associated with the early onset of coronary disease, kidney failure and stroke. Essential hypertension is generally asymptomatic and has been termed a silent killer.
  • the renin-angiotensin system plays an important role in salt-water homeostasis and the maintenance of vascular tone; stimulation or inhibition of this system respectively raises or lowers blood pressure, and may be involved in the etiology of hypertension. Hall, J.E., and Guyton, A.C. (1990), In Hypertension : Pathophysiology Diagnosis and Management, (Raven Press, Ltd., New York), pp.1105-1129.
  • Angiotensin converting enzyme (ACE) plays an important role in the renin-angiotensin system.
  • ACE acts on angiotensin I (Asp-Arg-Val-Tyr-Ile-His-Pro-Phe-His-Leu) , which is formed by decomposition of angiotensin secreted by the liver, by an enzyme, renin, produced in the kidney, and converts it to angiotensin II (Asp-Arg-Val-Tyr-Ile-His-Pro- Phe) .
  • Angiotensin II increases blood pressure by contracting the smooth muscles of the blood vessel walls and promoting secretion of aldosterone by action on the adrenal cortex.
  • ACE acts to create the blood pressure increasing enzyme of angiotensin II.
  • ACE decomposes and inactivates a protein called bradykinin, dilates the blood vessels and lowers blood pressure. Therefore, a common method of decreasing blood pressure in an individual is to inhibit the activity of ACE.
  • ACE inhibiting substances are known and commonly used for the purpose of decreasing blood pressure in patients.
  • ACE inhibiting pharmaceuticals is the synthetic chemical product known as captopril (D-2-methyl-3- mercaptopropanoyl-L-proline) , which is an oral hypotensive agent.
  • captopril D-2-methyl-3- mercaptopropanoyl-L-proline
  • anti-hypertensive agents are not without side effects such as the elevation of blood lipids and glucose. The elevation of blood lipids and glucose by these agents has been suggested as a reason why anti-hypertensive agents have not demonstrated any benefit to patients being monitored in death rate studies.
  • ACE inhibiting substances may be derived from casein and corn seed proteins (See Susumu Maruyama, Biosciences and Industry 47 (No. 11), 38-42
  • peptides do not have strong anti- hypertensive effects even if the peptides have strong ACE inhibition activity. Also, some peptides exhibit anti- hypertensive activity without exhibiting ACE inhibitory activity. Therefore, ACE inhibition is but one potential pharmacological mechanism of many of these peptides, or alternatively, the peptides have to be metabolized to other active forms which reduce blood pressure by ACE inhibition. Therefore, there is significant interest in obtaining anti-hypertensive peptides which can be produced in an industry stable manner and amount and which are effective in oral dosage and have low toxicity and great safety without appreciable side effects associated with existing anti- hypertensive pharmaceutical products.
  • an object of the present invention to provide novel anti-hypertensive peptides to be used for pharmaceutical compositions, dietary supplements, food ingredients, and foods for specified health uses of reducing and inhibiting hypertension and disease states associated with hypertension at a low cost with no appreciable side effects .
  • a further object of the present invention is to provide an anti-hypertensive composition comprising an anti- hypertensive peptide or its acceptable acid addition salt or base salt thereof, together with a pharmaceutically suitable diluent .
  • the present invention provides an anti-hypertensive peptide selected from the group of peptides consisting of Asp-Lys-Pro, Tyr-Lys-Pro, Glu-Lys-Pro, Asp-Ala-Pro, Glu-Ala- Pro, and His-Pro-Pro.
  • the present invention provides an anti- hypertensive agent comprising an effective amount of a peptide selected from the group consisting of Asp-Lys-Pro, Tyr-Lys-Pro, Glu-Lys-Pro, Asp-Ala-Pro, Glu-Ala-Pro, and His- Pro-Pro or a pharmacologically acceptable acid addition salt thereof .
  • the present invention provides a food composition suitable for treatment or prophylaxis of hypertension comprising a nutritional substance and a peptide selected from the group consisting of Asp-Lys-Pro, Tyr-Lys-Pro, Glu-Lys-Pro, Asp-Ala-Pro, Glu-Ala-Pro, and His- Pro-Pro or a pharmacologically acceptable acid addition salt thereof in an anti-hypertensively effective amount.
  • Yet a further object of the present invention is to provide methods of treating or prophylaxis of essential hypertension as well as hypertension associated with myocardial infarction, left ventricular systolic dysfunction, renal impairment or failure, congestive heart failure, and diabetes mellitus which comprises administering an effective amount of an anti-hypertensive peptide in a food or pharmaceutical composition.
  • A represents alanine
  • R represents arginine
  • N represents asparagine
  • D represents aspartic acid
  • C represents cysteine
  • Q represents glutamine
  • E represents glutamic acid
  • G represents glycine
  • H histidine
  • I represents isoleucine
  • L represents leucine
  • K represents lysine
  • M represents methionine
  • F represents phenylalanine
  • P represents proline
  • S represents serine
  • T represents threonine
  • Y represents tyrosine
  • V represents valine.
  • ACE angiotensin converting enzyme
  • treatment relate to any treatment of hypertensive disease or diseases related to hypertensive and include: (1) preventing hypertension from occurring in a subject who may be predisposed to the disease but who has not yet been diagnosed as having it; (2) inhibiting the disease, i.e., arresting its development; or (3) ameliorating or relieving the symptoms of ' he disease, i.e., causing regression of the hypertensive state.
  • Diseases related to hypertension include, but are not limited to, myocardial infarction, left ventricular systolic dysfunction, renal impairment or failure, congestive heart failure and diabetes mellitus.
  • anti-hypertensively effective amount shall mean an amount sufficient for treatment and prophylaxis of hypertension; an anti- hypertensively effective amount of the peptide of this invention would be an amount necessary to reduce, inhibit, or prevent hypertension in a subject.
  • a protein or polypeptide is considered substantially pure when that protein makes up greater than about 50% of the total protein content of the composition containing that protein, and typically, greater than about 60% of the total protein content. More typically, a substantially pure protein will make up from about 75 to about 90% of the total protein. Preferably, the protein will make up greater than about 90%, and more preferably, greater than about 95% of the total protein in the composition.
  • the terms "ingestible composition” are defined as foodstuffs and pharmaceutical products and preparations.
  • Typical ingestible compositions which include foodstuffs and pharmaceutical preparations of the present invention, are, for example, beverages, (including soft drinks, carbonated beverages, ready to mix beverages and the like), infused foods (e.g. fruits and vegetables), sauces, condiments, salad dressings, fruit juices, syrups, desserts (including puddings, gelatin, icings and fillings, baked goods and frozen desserts, such as ice creams and sherbets) , chocolates, candies, soft frozen products (such as soft frozen creams, soft frozen ice creams and yogurts, soft frozen toppings, such as dairy or non-dairy whipped toppings) , oils and emulsified products (such as shortening, margarine, mayonnaise, butter, cooking oil, and salad dressings), intermediate moisture foods, (e.g. rice and dog foods) and the like.
  • beverages including soft drinks, carbonated beverages, ready to mix beverages and the like
  • infused foods e.g. fruits and vegetables
  • sauces condiments,
  • Figure 1 is a graph representing the effects of oral doses of Asp-Lys-Pro and Tyr-Lys-Pro on Systolic Blood Pressure (Tail Cuff Method) administered to spontaneous hypertensive rats as discussed in Example 8 for in vivo dose response studies.
  • Figure 2 is a graph representing the effects of oral doses of His-Pro-Pro on Systolic Blood Pressure (Tail Cuff Method) administered to spontaneous hypertensive rats as discussed in Example 8 for in vivo dose response studies.
  • Figure 3 is a graph representing the effects of oral doses of Glu-Lys-Pro on Systolic Blood Pressure (Tail Cuff Method) administered to spontaneous hypertensive rats as discussed in Example 8 for in vivo dose response studies.
  • Figure 4 is a graph representing the effects of oral doses of Glu-Ala-Pro and Gly-Lys-Pro on Systolic Blood Pressure (Tail Cuff Method) administered to spontaneous hypertensive rats as discussed in Example 8 for in vivo dose response studies .
  • Figure 5 is a graph representing the effects of oral doses of Asp-Ala-Pro on Systolic Blood Pressure (Tail Cuff Method) administered to spontaneous hypertensive rats as discussed in Example 8 for in vivo dose response studies.
  • These anti-hypertensive peptides include Asp-Lys-Pro, Tyr-Lys-Pro, Glu-Lys-Pro, Asp-Ala-Pro, Glu-Ala-Pro, and His- Pro-Pro. Because these peptides have significant anti- hypertensive activity, they are also useful in the treatment and prophylaxis of hypertension, left ventricular systolic dysfunction, myocardial infarction, diabetes mellitus and progressive renal impairment or failure as well as other diseases caused by or associated with hypertension.
  • the peptides of the present invention may be either chemically synthesized or derived from food protein digest products. The method of action of these peptides remains unknown.
  • the mechanism of action may be ACE inhibition, or alternatively the peptides may have to be metabolized to other active forms which reduce blood pressure by ACE inhibition.
  • Other pharmaceutical methods of action may cause the anti-hypertensive activity of these peptides .
  • the present invention also provides pharmaceutical compositions comprising such anti-hypertensive peptides Asp- Lys-Pro, Tyr-Lys-Pro, Glu-Lys-Pro, Asp-Ala-Pro, Glu-Ala-Pro, and His-Pro-Pro or pharmacologically acceptable acid addition salts or base salts thereof.
  • the salt may be a pharmaceutically acceptable salt, including an inorganic acid salt, such as hydrochlorate, sulfate or phosphate, or an organic acid salt, such as citrate, maleate, fumarate, tartarate or lactate.
  • acid addition salts include pharmacologically acceptable acid (inorganic acid or organic acid) addition salts, such as hydrochloride, hydrobromide, sulfate, nitrate, acetate, benzoate, maleate, fumarate, succinate, tartrate, citrate, oxalate, methanesulfonate, tolunesulfonate, aspartate, glutamate, etc.
  • alkali and alkaline earth metal and ammonium salts include pharmacologically acceptable salts, such as sodium, potassium, calcium and ammonium salts
  • organic base salts include pharmacologically acceptable salts, such as basic amino acid salts including lysine and ornithine salts.
  • the anti-hypertensive peptides of the present invention may be synthesized using either solid-phase peptide synthesis, by classical solution peptide synthesis, as well as by liquid-phase peptide synthesis using a soluable oligomeric support.
  • Val-Pro-Pro Using Val-Pro-Pro, Enalapril and Lisinopril as starting templates, several series of peptide analogs such as X-Pro-Pro, X-Ala-Pro, and X-Lys-Pro, wherein X represents any amino acid residue, may be synthesized using solid-phase peptide synthesis, classic solution peptide synthesis, or oligomer-supported liquid-phase peptide' synthesis.
  • Classic solution peptide synthesis methods may be used with traditional purification steps and without the use of any polymer as a basis of attachment for the products as a method of separating the product from the unreacted reactants.
  • the product from each step of the synthesis can then be separated from unreacted reactants based on the large difference in size between the relatively large polymer- attached product and the unreacted reactants. This permits reactions to take place in homogeneous solutions, and homogeneous solutions, and eliminates tedious purification steps associated with classic solution phase synthesis.
  • the procedure entails the sequential assembly of the appropriate amino acids into a peptide of a desired sequence while the end of the growing peptide is linked to an insoluble support.
  • the carboxyl terminus of the peptide is linked to a polymer from which it can be liberated upon treatment with a cleavage reagent .
  • an amino acid is bound to a resin particle, and the peptide generated in a stepwise manner by successive additions of protected amino acids to produce a chain of amino acids. Modifications of the technique described by Merrifield are commonly used. See, e.g., Merrifield, J. Am. Chem. Soc.
  • peptides are synthesized by loading the carboxy-terminal amino acid onto an organic linker, which is covalently attached to an insoluble polystyrene resin cross-linked with divinyl benzene. Synthesis is accomplished in an automated peptide synthesizer, such as that available from Applied Biosystems . See, e.g., Model 430-A, Applied Biosystems, Foster City, Calif. Following synthesis, the product may be removed from the resin. A routine synthesis may produce 0.5 mmole of peptide-resin. Following cleavage and purification, a yield of approximately 60 to 70% is typically produced.
  • Purification of the product peptides is accomplished by, for example, crystallizing the peptide from organic solvents or reverse high-pressure liquid chromatography (e.g., using a C. sup.18 column with 0.1% trifluoroacetic acid and acetonitrile as solvents) . Purified peptide may be lyophilized and stored in a dry state until use. Analysis of the resulting peptides may be accomplished using the common methods of analytical high pressure liquid chromatography (HPLC) and electrospray mass spectro etry (ES-MS) .
  • HPLC high pressure liquid chromatography
  • ES-MS electrospray mass spectro etry
  • Acid addition salts of the present peptides can be prepared according to a conventional method.
  • an acid addition salt can be obtained by reacting one of the present peptides containing a basic amino acid residue with a suitable acid in one equivalent amount thereto in water and then freeze-drying the product.
  • alkali or alkaline earth metal salts, ammonium salts or organic base salts (hereinafter these are referred to as base salts) can also be prepared according to a conventional method.
  • a base salt can be obtained by reacting one of the present peptides containing an acidic amino acid residue with a suitable base in one equivalent amount thereto in water and then freeze-drying the product.
  • the present peptides and their acid addition salts and base salts have a hypotensive activity, and thus are effective for treatment and/or prophylaxis of hypertension of mammals including human beings.
  • present peptides and their acid addition salts or base salts are used alone or in the form of a food or pharmaceutical composition including at least one nutritional or pharmaceutical auxiliary.
  • the present peptides and their acid addition salts and base salts can be administered parenterally, such as by intravenous injection or rectal administration, or orally, and formulated into a form suitable for each administration method.
  • compositions for injections typically include sterilized aqueous solutions.
  • Such formulations of the above form can further contain pharmaceutical auxiliaries other than water such as a buffering and a pH adjusting agent (for example, sodium hydrogenphosphate, citric acid) , a tonicity agent (e.g., sodium chloride, glucose), or a preservative (methyl p-hydroxybenzoate, propyl-hydroxybenzoate or the like) .
  • a buffering and a pH adjusting agent for example, sodium hydrogenphosphate, citric acid
  • a tonicity agent e.g., sodium chloride, glucose
  • preservative methyl p-hydroxybenzoate, propyl-hydroxybenzoate or the like
  • the formulations can also be prepared as a sterilizing solid composition and dissolved at the time of use in sterilized water to form a solution for administration to the subject.
  • Orally administered agents are prepared in a form suitable for absorption in gastrointestinal organs.
  • Food compositions contain at least one nutritional substance as a food auxiliary.
  • Tablets, capsules, granules, fine granules and powders can contain conventional pharmaceutical auxiliaries, such as, a binder (e.g., syrup, gum arabic, gelatin, sorbitol, tragacanth, polyvinylpyrrolidone or hydroxycellulose) , an excipient (e.g., lactose, sucrose, corn starch, calcium stearate, sorbitol or glycine) , a lubricant (e.g., magnesium stearate, talc, polyethylene glycol or silica) , a disintegrant (potato starch or carboxymethylcellulose) , or a wetting agent (e.g., sodium lauryl sulfate) .
  • a binder e.g., syrup, gum arabic, gelatin, sorbitol, tragacanth, polyvinylpyrrolidone or hydroxycellulose
  • an excipient e.g.,
  • Oral liquid agents can be aqueous solutions or the like, or dry products to be dissolved at the time of use to create a solution.
  • Such oral liquid agents may contain conventional additives such as a preservative (methyl p-hydroxybenzoate, propyl p- hydroxybenzoate, sorbic acid or the like) .
  • the anti-hypertensive peptides and their acid addition salts or base salts can also be ingested as an additional ingredient contained in foods and drinks, or as part of a functional food or health food.
  • these peptides can be formulated together with nutrients such as various vitamins and minerals, into liquid foods such as nutrient drinks, soy milks and soups or solid foods of various types, or used in the form of a powder or by incorporation into various foods.
  • the content of the effective anti- hypertensive ingredient in such a functional or health food can be similar to the dose contained in a typical pharmaceutical agent.
  • the amount of the present peptide or an acid addition salt or base salt thereof in the present anti-hypertensive agent can be varied, but it is preferred that the amount of peptide or acid addition salt or base salt in the agent be 1 to 100% (w/w) , preferably 10 to 100% (w/w) .
  • the preferred amount for administration of the effective ingredients of the present anti-hypertensive agent or ACE inhibitor is 1 to 200 mg/kg/day.
  • the preferred dosage is 1 to 200 mg/kg/day for the administration of the peptides of this invention for the method of inhibition of ACE and for the method of treatment or prophylaxis of hypertension.
  • the tripeptides of the present invention are synthesized according to the procedure outlined below.
  • the following examples are intended to illustrate but not limit the present invention.
  • Aspartyl-Alanyl-Proline (0.62 g, 80% isolated yield) was prepared using substantially the same procedure as described in Example 1.
  • Step 1 Preparation of N ⁇ -Boc-N ⁇ -di-Boc-Histidyl-Prolyl-Proline
  • Step 2 Preparation of Histidyl-Prolyl-Proline A solution of 4 N HCl in 1,4-dioxane (10 mL) was added to a solution of Boc-His (Boc) -Pro-Pro in the same solvent (10 mL, anhydrous) . The mixture was stirred at room temperature under N 2 for 1 hour. The volatile was removed under reduced pressure. The resulting yellow oil was then purified by prep-HPLC to obtain 2TFA. His-Pro-Pro (470 mg, 32.6% overall yield) .
  • Step 2 Preparation of Tyrosyl-Lysyl-Proline
  • H-Lys (Boc) -Pro-OtBu 1.7 mmol
  • Boc-Tyr-OH 0.8 g, 1.7 mmol
  • DMF 10 mL
  • HOBt 0.26 g, 2 mmol
  • HBTU 0.76 g, 2 mmol
  • DIEA 0.52 g, 4 mmol
  • Boc-Asp-Lys (Boc) -Pro-OtBu was prepared by prep-HPLC and lyophilized.
  • Step 2 Preparation of Glutamyl-Lysyl-Proline
  • H-Lys (Boc) -Pro-OtBu 1.7 mmol
  • Boc-Glu-OH 1.7 mmol
  • DMF 10 mL
  • HOBt 0.26 g, 2 mmol
  • HBTU 0.76 g, 2 mmol
  • DIEA 0.52 g, 4 mmol
  • Germantown, New York, 6 animals per group, about 500g each) were used as test animals to examine the anti-hypertensive effects of intravenously administered synthetic peptides such as X-Pro-Pro, X-Ala-Pro, and X-Lys-Pro, wherein X represents any amino acid residue.
  • Catheters were placed in the femoral veins of the animals for intravenous infusion.
  • Example 8 Dose Response Studies Dose response studies of the tripeptides Asp-Lys-Pro, Asp- Ala-Pro, Tyr-Lys-Pro, Glu-Ala-Pro, Glu-Lys-Pro, His-Pro-Pro, and Gly-Lys-Pro were performed on spontaneous hypertensive rats (SHR) with six (6) rats in each test group.
  • SHR spontaneous hypertensive rats
  • Each testing peptide was administered in dosages of 0, 0.3, 1, 3, 10, and 30 umols/rat of each as well as lOumols/rat of Enalapril as a positive control in a group of six (6) rats, and the blood pressure was measured in these SHR at four (4) hours after oral administration of these peptides and Enalapril by tail cuff method using a noninvasive blood pressure apparatus (Twelve channel Semi-Automatic NIBP Test System, IITC Inc.) . The doses were delivered orally in about 1 ml volume. The result of this study was that all of the tripeptides administered significantly decreased blood pressure in these SHR. The maximal blood pressure lowering observed was about 25-30 mmHg. Graphic results of this study can be seen in Figures 1-5. The results of these tests for the peptides of this invention are shown in the following Tables . Table 2

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Organic Chemistry (AREA)
  • Molecular Biology (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • Mycology (AREA)
  • Nutrition Science (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Immunology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

L'invention concerne de nouveaux tri-peptides qui présentent une activité hypotensive. Ces peptides peuvent être utilisés comme ingrédients actifs dans des compositions pharmaceutiques, des suppléments diététiques et des ingrédients alimentaires. Cette invention a également trait à l'utilisation de ces nouveaux peptides antihypertenseurs destinés à des méthodes de traitement et de prophylaxie d'hypertension primaire ainsi que d'hypertension liée à un infarctus du myocarde, un dysfonctionnement systolique du ventricule gauche, un diabète sucré, une insuffisance rénale progressive et une insuffisance cardiaque congestive.
PCT/US2001/007530 2000-03-10 2001-03-09 Nouveaux peptides a activite hypotensive WO2001068114A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU2001247334A AU2001247334A1 (en) 2000-03-10 2001-03-09 Novel peptides with anti-hypertensive activity

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US18849700P 2000-03-10 2000-03-10
US60/188,497 2000-03-10

Publications (1)

Publication Number Publication Date
WO2001068114A1 true WO2001068114A1 (fr) 2001-09-20

Family

ID=22693397

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2001/007530 WO2001068114A1 (fr) 2000-03-10 2001-03-09 Nouveaux peptides a activite hypotensive

Country Status (2)

Country Link
AU (1) AU2001247334A1 (fr)
WO (1) WO2001068114A1 (fr)

Cited By (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002062828A2 (fr) * 2001-02-05 2002-08-15 Neurotell Ag Tripeptides derives de tripeptides utilises dans le traitement d'affections neurodegeneratives
WO2004022083A1 (fr) * 2002-09-04 2004-03-18 Dsm Ip Assets B.V. Composition nutritionnelle et therapeutique d'agent sensibilisant a l'insuline et de fraction peptidique
WO2004087743A2 (fr) * 2003-03-31 2004-10-14 Council Of Scientific And Industrial Research Molecules contre l'hypertension et leur procede de preparation
WO2005058070A1 (fr) * 2003-12-15 2005-06-30 Unilever N.V. Peptides presentant un effet inhibiteur de l'eca
US6994987B1 (en) 1999-11-11 2006-02-07 Calpis Co., Ltd. Process for producing tripeptides
US7029670B2 (en) 2000-12-28 2006-04-18 Calpis Co., Ltd. Medicines for relieving intestinal disorders
US7122524B2 (en) 2001-02-05 2006-10-17 Neurotell Ag Tripeptides and tripeptide derivatives for the treatment of postlesional disease of the nervous system
WO2006114441A1 (fr) * 2005-04-28 2006-11-02 Dsm Ip Assets B.V. Hydrolysats proteiques abaissant la pression arterielle
US7163922B2 (en) 2001-02-05 2007-01-16 Neurotell Ag Tripeptide derivatives for the treatment of postlesional diseases of the nervous system
US7279971B2 (en) 2002-09-06 2007-10-09 Telefonaktiebolaget Lm Ericsson (Publ) Composite power amplifier
US7282354B2 (en) 1997-09-26 2007-10-16 Calpis Co., Ltd. Method for producing fermented milk product
US7759108B2 (en) 1999-01-11 2010-07-20 Calpis Co., Ltd. Method for producing fermented milk containing angiotensin converting enzyme inhibitory peptide and method for producing whey
US7972808B2 (en) 2002-06-04 2011-07-05 Dsm Ip Assets B.V. Protein hydrolysate rich in tripeptides
CN110734470A (zh) * 2019-11-18 2020-01-31 浙江省农业科学院 一种降血脂的蚕蛹蛋白肽hpp及其应用
CN117534727A (zh) * 2023-12-21 2024-02-09 中国药科大学 一种食源性降压肽的制备方法和应用

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5691310A (en) * 1987-09-29 1997-11-25 Vesely; David L. Methods of treatment using proANF peptides
US5854029A (en) * 1996-08-02 1998-12-29 Calpis Co., Ltd. Method for preparing antihypertensive agent

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5691310A (en) * 1987-09-29 1997-11-25 Vesely; David L. Methods of treatment using proANF peptides
US5854029A (en) * 1996-08-02 1998-12-29 Calpis Co., Ltd. Method for preparing antihypertensive agent

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
CASARINI ET AL.: "Angiotensin converting enzymes from human urine of mild hypertensive untreated patients resemble the n-terminal fragment of human angiotensin I-converting enzyme", INT. J. BIOCHEM. CELL BIOLOGY, vol. 33, January 2001 (2001-01-01), pages 75 - 85, XP002944569 *
FIAT ET AL.: "Further data on the specificity of renin: renin-sensitive sequence of sheep kappa-casein", CHIMIA, vol. 24, 1970, pages 220, XP002944567 *
MONTECUCCHI ET AL.: "Primary structure determination of a tryptophan-containing tridecapeptide from phyllomedusa rohdei", INT. J. PEPTIDE PROTEIN RES., vol. 27, 1986, pages 175 - 182, XP002944565 *
NAKAMURA ET AL.: "Antihypertensive effect of sour milk and peptides isolated from it that are inhibitors to angiotensin I-converting enzyme", J. DIARY SCI., vol. 78, 1995, pages 1253 - 1257, XP002944568 *
ZAIDI ET AL.: "Amino acid sequence of a peptide from the hypothalamus of rana cyanophlyctis", INT. J. PEPTIDE PROTEIN RES., vol. 18, 1981, pages 516 - 518, XP002944566 *

Cited By (22)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7282354B2 (en) 1997-09-26 2007-10-16 Calpis Co., Ltd. Method for producing fermented milk product
US7759108B2 (en) 1999-01-11 2010-07-20 Calpis Co., Ltd. Method for producing fermented milk containing angiotensin converting enzyme inhibitory peptide and method for producing whey
US6994987B1 (en) 1999-11-11 2006-02-07 Calpis Co., Ltd. Process for producing tripeptides
US7029670B2 (en) 2000-12-28 2006-04-18 Calpis Co., Ltd. Medicines for relieving intestinal disorders
US7129213B2 (en) 2001-02-05 2006-10-31 Neurotell Ag Tripeptides and tripeptide derivatives for the treatment of neurodegenerative diseases
US7122524B2 (en) 2001-02-05 2006-10-17 Neurotell Ag Tripeptides and tripeptide derivatives for the treatment of postlesional disease of the nervous system
WO2002062828A2 (fr) * 2001-02-05 2002-08-15 Neurotell Ag Tripeptides derives de tripeptides utilises dans le traitement d'affections neurodegeneratives
US7163922B2 (en) 2001-02-05 2007-01-16 Neurotell Ag Tripeptide derivatives for the treatment of postlesional diseases of the nervous system
WO2002062828A3 (fr) * 2001-02-05 2002-10-10 Tell Pharm Ag Tripeptides derives de tripeptides utilises dans le traitement d'affections neurodegeneratives
US7972808B2 (en) 2002-06-04 2011-07-05 Dsm Ip Assets B.V. Protein hydrolysate rich in tripeptides
WO2004022083A1 (fr) * 2002-09-04 2004-03-18 Dsm Ip Assets B.V. Composition nutritionnelle et therapeutique d'agent sensibilisant a l'insuline et de fraction peptidique
US7279971B2 (en) 2002-09-06 2007-10-09 Telefonaktiebolaget Lm Ericsson (Publ) Composite power amplifier
WO2004087743A3 (fr) * 2003-03-31 2005-05-19 Council Scient Ind Res Molecules contre l'hypertension et leur procede de preparation
WO2004087743A2 (fr) * 2003-03-31 2004-10-14 Council Of Scientific And Industrial Research Molecules contre l'hypertension et leur procede de preparation
US7335644B2 (en) 2003-03-31 2008-02-26 Council Of Scientific And Industrial Research Anti-hypertensive molecules and process for preparation thereof
AU2004298742B2 (en) * 2003-12-15 2008-07-31 Unilever Plc Peptides having an ace inhibiting effect
WO2005058070A1 (fr) * 2003-12-15 2005-06-30 Unilever N.V. Peptides presentant un effet inhibiteur de l'eca
EA012972B1 (ru) * 2005-04-28 2010-02-26 ДСМ АйПи АССЕТС Б.В. Трипептиды мар и itp или их соли, белковые гидролизаты и смеси, содержащие указанные трипептиды или их соли, для снижения кровяного давления
WO2006114441A1 (fr) * 2005-04-28 2006-11-02 Dsm Ip Assets B.V. Hydrolysats proteiques abaissant la pression arterielle
CN110734470A (zh) * 2019-11-18 2020-01-31 浙江省农业科学院 一种降血脂的蚕蛹蛋白肽hpp及其应用
CN117534727A (zh) * 2023-12-21 2024-02-09 中国药科大学 一种食源性降压肽的制备方法和应用
CN117534727B (zh) * 2023-12-21 2024-04-19 中国药科大学 一种食源性降压肽的制备方法和应用

Also Published As

Publication number Publication date
AU2001247334A1 (en) 2001-09-24

Similar Documents

Publication Publication Date Title
JP4369985B2 (ja) アンジオテンシン変換酵素阻害ペプチド
JP4669396B2 (ja) アンジオテンシン変換酵素阻害剤、医薬及び機能性食品
WO2001068113A1 (fr) Peptides antihypertenseurs
WO2001068114A1 (fr) Nouveaux peptides a activite hypotensive
JP5832049B2 (ja) ジペプチジルペプチダーゼ−ｉｖ阻害剤
JP2007523940A (ja) 抗高血圧ペプチド
ES2281567T3 (es) Nuevo peptido que tiene efecto inhibidor de angiotensina convertasa.
JP5417405B2 (ja) アンジオテンシン変換酵素阻害性降圧ペプチド組成物の製造方法
JP4628958B2 (ja) アンジオテンシン変換酵素阻害ペプチド
JP5416964B2 (ja) 新規トリペプチドおよびそれらトリペプチドの製造法、ならびにアンジオテンシン変換酵素阻害物質の製造方法
JPWO2013133032A1 (ja) ジペプチジルペプチダーゼ−ｉｖ阻害剤
JP5877560B2 (ja) ジペプチジルペプチダーゼ−ｉｖ阻害剤
JP3103363B2 (ja) 新規トリペプチド及び血圧降下剤
WO2007094340A1 (fr) Vasodilatateur
JP5175196B2 (ja) アミノペプチダーゼｐのチオ含有インヒビター、その組成物および使用法
JP2866872B2 (ja) 血圧降下剤

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BY BZ CA CH CN CR CU CZ DE DK DM DZ EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NO NZ PL PT RO RU SD SE SG SI SK SL TJ TM TR TT TZ UA UG US UZ VN YU ZA ZW

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): GH GM KE LS MW MZ SD SL SZ TZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE TR BF BJ CF CG CI CM GA GN GW ML MR NE SN TD TG

121 Ep: the epo has been informed by wipo that ep was designated in this application
REG Reference to national code

Ref country code: DE

Ref legal event code: 8642

122 Ep: pct application non-entry in european phase
NENP Non-entry into the national phase

Ref country code: JP