WO1993011799A1 - Soybean protein or hydrolyzates in pharmaceutical compositions to protect bioactive peptides from enzymatic inactivation - Google Patents
Soybean protein or hydrolyzates in pharmaceutical compositions to protect bioactive peptides from enzymatic inactivation Download PDFInfo
- Publication number
- WO1993011799A1 WO1993011799A1 PCT/US1992/009336 US9209336W WO9311799A1 WO 1993011799 A1 WO1993011799 A1 WO 1993011799A1 US 9209336 W US9209336 W US 9209336W WO 9311799 A1 WO9311799 A1 WO 9311799A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- protein
- protecting agent
- molecular weight
- therapeutic agent
- agent
- Prior art date
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/81—Protease inhibitors
- C07K14/8107—Endopeptidase (E.C. 3.4.21-99) inhibitors
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/22—Hormones
- A61K38/2207—Gastrins; Cholecystokinins [CCK]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/33—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans derived from pro-opiomelanocortin, pro-enkephalin or pro-dynorphin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
Definitions
- This invention relates to enhancing the bioavailability of proteolytically labile therapeutic agents by administering the therapeutic agent in combination with a protecting agent comprising a protein, a purified natural protein, a molecular weight fractionated protein, or a partially hydrolyzed protein.
- Peptide drugs and drugs containing a peptidase labile bond are among the most promising medicinal agents of modern times, but their instability in the presence of proteolytic enzymes in the gastrointestinal tract and other mucosal tissues usually requires that they be administered parenteraliy. Although patients can be taught to inject parenteraliy, there has been a long felt need to develop a non-invasive method for self administration of peptide drugs.
- Protease inhibitors and penetration enhancers are means often considered to circumvent the enzymatic and penetration barriers to peptide and protein absorption from mucosal routes of administration. Because of such barriers, the bioavailability of peptide and protein drugs from mucosal routes is poor.
- Non-parenteral administration of peptide drugs in particular often results in very low bioavailability because of hydrolysis of the peptides by proteolytic enzymes.
- Lee Journal of Controlled Release, 13, 213 (1990).
- proteolytic enzymes which inactivate proteolytically-labile therapeutic agents are pepsin, trypsin, chymotrypsin, elastase, and carboxypeptidase in the intestinal lumen, and the aminopeptidases located on the mucosal surfaces of the Gl tract, nose, and vagina.
- Kidron, et al; US 4,579,730 (1986) disclose the use of protease inhibitors in oral formulation of insulin.
- Soybean flour is disclosed as a source of soybean trypsin inhibitor (Bowman-B ⁇ rktrypsin/chymotrypsin inhibitor; molecular weight 8000 daltons).
- Ziv, et al; B ⁇ ochem. Pharmacol. 36, 1035-1039 (1987) disclose the use of the protease inhibitor aprotinin to enhance the oral absorption of proteins.
- Losse, et al; East German Patent DD 252 539 A1 (1987) disclose the use of eps ⁇ on-aminocaproic acid and aprotinin as protease inhibitors in oral formulation of peptides.
- ⁇ Lee; J.; Controlled Release 13, 213-223 (1990) reviews the use of protease inhibitors in formulations of peptides for oral, nasal, buccal, rectal, vaginal, pulmonary, and ocular routes.
- An alkaline material such as sodium sutfite, sodium carbonate or sodium hydroxide is used to extract glycinin at a pH 6.4-6.8.
- the glycinin is then precipitated from the extract (e.g., pH 4.2-4.6) at its isoelectric pH in which sulfur dioxide may be utilized as the adjusting acid.
- the precipitated glycinin product is then modified with pepsin under temperature and pH conditions conducive to hydrolysis of protein.
- the glycinin is hydrolyzed with pepsin until its water-solubility is increased to 40-50%.
- U.S. Patent No. 2,502,482 by Sair et al. reports the enzymatic modification of glycinin with pepsin to produce an isolate wherein at least 60% by weight of the pepsin modified isolate is water-soluble at a pH 5.0.
- Puski reports the enzymatic modifying of soy isolates (precipitated at pH 4.5) with Aspergillus oryzae in "Modification of Functional Properties of Soy Proteins by Proteolytic Enzyme Treatment” (Cereal Chem. 52, pages 655-665 (1975)).
- protecting agents enhance the oral, nasal, rectal and vaginal bioavailability of proteolytically-labile therapeutic agents which, in the absence of the protecting agents, would suffer enzymatic inactivation upon attempted oral, nasal, rectal or vaginal administration.
- This invention comprises a protecting agent and a pharmaceutically effective amount of a proteolytically-labile therapeutic agent, with the proviso that when said protecting agent is soy flour said therapeutic agent may not be insulin.
- this invention comprises a method of enhancing bioavailability of a proteolytically-labile therapeutic agent to a mammal or other animal in need of said therapeutic agent comprising administering said therapeutic agent in combination with a bioavailability enhancing amount of protecting agent with the proviso that when said protecting agent is soy flour said therapeutic agent may not be insulin.
- a proteolytically-labile therapeutic agent which is an active ingredient in a pharmaceutical composition of the invention, includes those which possess peptide bonds in their structure or which, upon exposure to various proteolytic enzymes present in the digestive tract or nasal or vaginal mucosa, are inactivated by decomposition, denaturation or other means. When administered orally, nasally, rectally or vaginally, therefore, the active therapeutic agents cannot be absorbed or cannot produce their therapeutic effects to a satisfactory extent.
- proteolytically-labile therapeutic agents examples include peptides, such as calcitonin, prolactin, adrenocorticotropin, thyrotropin, growth hormone, gonadotropic hormone, oxytocin, vasopressin, gastrin, tetragastrin, pentagastrin, glucagon, secretin, pancreozymin, substance P and gonadotropin.
- proteolytically-labile therapeutic agents are luteinizing releasing hormone, leuprolide, enkephalin, follicle stimulating hormone, cholecystokinin.thymopentin, endothelin, neurotensin, interferon, interleukins, insulin, and insulinotropin.
- proteolytically-labile therapeutic agents there may also be used purified extracts of natural origin and their chemical modifications as well as products obtained by tissue culture and products obtained by cultivating microorganisms or cells rendered productive by genetic engineering techniques.
- the proteolytically-labile therapeutic agents may also include synthetic peptides and derivatized synthetic peptides such as Terlakiren(lsopropyl-N-[N-(4-mo ⁇ holine-carbonyl)-L-phenylalanine-S-methyl-cystein ⁇ ]-
- Protecting agents of the present invention may be chemically synthesized proteins and peptides, natural proteins, purified natural proteins, chemically modified natural proteins or partially hydrolyzed natural proteins, or proteins which have been fractionated according to molecular weight, polarity, or charge, or mixtures thereof. Natural, food-grade proteins or partially hydrolyzed food-grade proteins are preferred.
- the molecular weight of the protecting agent should be greater than 1000.
- the procedures for molecular weight fractionation of the protecting agent may be varied to produce any desired molecular weight fraction of a natural protein.
- Solvent extraction may be used to separate proteins according to molecular weight, polarity or charge.
- Enzymatic or chemical hydrolysis of a protein may be followed by separation of the desired molecular weight fraction by ultrafiltration membranes or dialysis membranes.
- Molecular weight fractionation may also be effected by gel chromatography or other means.
- Hydrolysis of proteins or peptides may be carried out by heat treatment, or by treatment with acid or base or cyanogen bromide or by other chemical means.
- Enzymatic treatment may be carried out with a single proteolytic enzyme, or with various combinations of proteolytic enzymes, acting concurrently or sequentially.
- a variety of proteolytic enzymes may be used, including but not limited to trypsin, chymotrypsin, elastase, carboxypeptidase, aminopeptidase, pepsin, and collagenase.
- Proteinaceous materials Naturally occurring proteins of animal or vegetable origin are preferred.
- proteinaceous starting materials include but are not limited to soy flour, soy protein, wheat gluten, almond powder, peanut powder, casein, fish protein, and the like.
- the protecting agents of this invention function as sacrificial protease inhibitors which thereby enhance the bioavailability of pharmaceutical agents that are labile to certain proteases that can degrade the pharmaceutical agents upon oral, nasal, vaginal or rectal administration.
- Coadministration of these protecting agents with the labile pharmaceutical agents results in (1) competitive occupancy of the degrading proteases by said protecting agents, (2) inhibition of protease degradation of the pharmaceutical agents resulting in enhanced absorption and therapeutic effectiveness, and (3) ultimate metabolism and absorption of the protecting agents.
- the dissolution rate of the protecting agent it is believed to be desirable to match the dissolution rate of the protecting agent to the dissolution rate of the proteolytically-labile therapeutic agent.
- short dissolution times of the protecting agent are more effective for low molecular weight therapeutic agents.
- Peptides ranging in molecular weight of > 1000 to ⁇ 100,000 are preferred, with a molecular weight of >1000 to ⁇ 30,000 being especially preferred.
- the effective protecting agent fraction must be matched to the particular lability characteristics of the proteolytically-labile therapeutic agent.
- a protecting agent fraction which has a fast dissolution rate and is effective against aminopeptidase is preferred.
- a preferred protecting agent is the ⁇ 30,000 MW fraction of pepsin-treated decanted soy flour, as demonstrated in Example 15.
- a preferred protecting agent is one which improves the systemic bioavailability of the therapeutic agent when the protecting agent is dosed at a practical total dose, as described above.
- Preferred protecting agent fractions, for a particular proteolytically-labile therapeutic agent are obtained by the decanting, filtration, extraction, hydrolysis, and size fractionation processes described herein.
- Preferred protecting agent fractions for a particular proteolytically-labile therapeutic agent are identified utilizing in vitro and in vivo procedures such as those exemplified herein.
- the pharmaceutical composition according to the present invention is preferably administered to a mammal or other animal in need of such treatment in any form in which a protecting agent and proteolytically-labile therapeutic agent are allowed to coexist in the intestine, for example, in the form of tablets, granules or capsules, with both ingredients provided with an enteric coating either separately or compositely.
- the composition may also be administered rectally or vaginally in the form of suppositories prepared by adding both ingredients to a suppository base in ordinary use.
- the protecting agent and proteolytically-labile therapeutic agent may be dosed together in a nasal spray. Where desirable, these dosage forms may be added with various pharmaceutically acceptable additives such as excipients and emuls ' rfiers.
- the dose of the proteolytically labile therapeutic agent is preferably 0.0001 to
- the amount of the protecting agent will depend upon the route of administration, the lability of the therapeutic agent, and the dose of the therapeutic agent.
- the protecting agent will generally be dosed at about 10-1500 mg.
- the protecting agent will be dosed at 10 mg to about 15 gm.
- the dose of protecting agent will be generally lower, in the range about 1-100 mg.
- compositions for intestinal absorption according to this invention were evaluated with respect to their effectiveness, with the results given below.
- Soy Flour (from Sigma Chem. Co.), 4.5 g in 135 ml 0.01 M pH 7.5 phosphate buffer and 15 ml 0.05% Thimerosal was stirred for 15 minutes, sonicated for 10 minutes and agitated for 25 hours at room temperature. The mixture was allowed to settle, the supernatant drawn off, centrifuged, and filtered. In this way the directly-soluble fraction was obtained for recovery and use or further processing.
- Example 1 The procedure of Example 1 was followed to prepare a solution of decanted and filtered soy flour which was processed further as follows to achieve molecular weight
- the retentate (>100K) was discarded, and the permeate ( ⁇ 100K) was separated using a 30K nominal molecular weight Pellicon membrane.
- the second (30K-100K) was saved, and the second permeate was separated on a 1 K nominal molecular weight Pellicon membrane.
- the third retentate (1 K-30K) was saved, and the third permeate ( ⁇ 1 K) was discarded.
- the 1 K-30K fraction was freeze-dried, and the 30K-100K fraction was dried in a vacuum oven.
- the yield of the 1 K-30K fraction (7.33 g) was 2.5% of the starting soy flour.
- the yield of the 30K-100K fraction (5.25 g) was 1.8% of the starting soy flour.
- Soy flour was hydrolysed into lower molecular weight (MW) fragments using pepsin and the hydrolysate was fractionated into 1000-3500, 3500-6/8K and 6/8K- 12/14K MW fractions according to the following procedure.
- Soy flour (Sigma # S-99633; 5.4 g) and Thimerosal (50 pp in final concentra ⁇ tion) were added to 180 ml of a solution containing pH 1.9 0.2N KCI/0.2 N HCI and mixed for one-half hour after which Pepsin (Sigma #P-6887; 18.0 mg) was added. Aiiquots of 20 ml each were placed in each of 9 pieces of 12,000-14,000 Molecular Weight Cut-Off (MWCO) Spectrum dialysis tubing, and were dialyzed against 55 ml of pH 1.9 KCI buffer at 37° C in a shaking water bath. The buffer was changed after 2 hours and after 6 hours and dialysis was continued for 24 hours. Permeates ( ⁇ 12/14K) from each time period were combined and evaporated at 55° C in a vacuum oven.
- MWCO Molecular Weight Cut-Off
- the 2, 6 and 24 hour samples were placed in 1000 MWCO tubing and dialyzed against water.
- the resulting retentates (1K-12/14K) were evaporated at 55° C in a vacuum oven, giving a total weight of 449.5 mg; 418 mg of this material was dissolved in 30 ml deionized water and placed in two pieces of 3500 MWCO dialysis tubing, and dialyzed against 55 ml water at room temperature for 24 hours.
- the water was changed after 2, 6 and 24 hours dialysis, and the permeates (1 K-3.5K) were combined and evaporated at 55° C in a vacuum oven.
- Soy flour (600 mg) was dispersed in 20 ml 0.01 M potassium phosphate buffer, pH 7.5 with 50 ppm Thimerosal. 2 Mg of trypsin was added and the mixture was placed in 12/14K MWCO dialysis tubing. The mixture was dialysed against 50 ml of buffer at 37° C in a shaking water bath. The buffer was changed after 2 hours and 6 hours and the dialysis continued for 24 hours. 50 ppm Thimerosal was added to the 6 hour buffer. This procedure was carried out in triplicate.
- elastase 0.182 ml of elastase solution, 11 mg protein/ml
- the buffer was changed after 6 hours and dialysis was continued for 24 hours. After determining absorbance at 280 nm, the permeates were evaporated as described above.
- Material so-obtained was designated as sequentially-treated trypsin elastase soy, MW ⁇ 12/14K.
- Renin antagonist tripeptide terlakiren 200 mg of solid crystalline drug powder in a hard gelatin capsule formulation
- aqueous slurry 1 g of the test inhibitor in 150 ml water.
- Serum levels of tripeptide were measured a 6 time points post-dose: 15 min, 30 rnin, 1 hr, 2 hr, 3 hr and 4 hr.
- Four fasted dogs were used for each study, each serving as its own control on a preceding week. Serum was extracted with N-butyl chloride followed by incubation with an aqueous solution of chymotrypsin.
- the degradation product was assayed, after derivitization with fluorescamine.
- the fluorescence detector was a Kratos Spectroflow 280.
- the column was a Waters Novapak C-18.
- the emission wavelength was 380 nm.
- the mobile phase was 75:25 waterracetonitrile and flow rate 1.0 ml/min.
- the detection limit was 10 ng/ml.
- Zero-to-four-hour areas under curves (AUCs) were calculated from the concentration-vs-time plots for each dog using the trapezoidal rule.
- Table II demonstrates that commercially available soy protein (PP 620 from Protein Technologies Inc.) and a 1-30K fraction of processed soy flour (prepared as in Example 3) enhance the oral bioavailability of teriakerin, a chymotrypsin-labile therapeutic agent.
- test inhibitor at concentrations of about 0.1 to/or 0.5 mg/ml
- pH 6.5 citric acid (0.10 M)/disodium phosphate (0.20M) buffer at a final buffer concentration of 300 mOsm and incubated 37° C. Samples were taken at times
- HPLC analysis of teriakiren was carried out using a Waters Resolve 5u C-18 column.
- the mobile phase was a water:acetonitrile (50:50) mixture to which was added
- % inhibition 100 x [1-k ⁇ J where K ⁇ is the initial degradation rate of terlakiren in the presence of the protecting agents and k e is the initial degradation rate of teriakiren without protecting agents.
- Ki is defined as the inhibition Michaelis-Menton constant - a conventional measure of the affinity of an inhibitor for the active site and, hence, its potency as an inhibitor of the enzyme. Ki determinations can be carried out from the initial degradation rate data acquired at several concentrations of inhibitor at constant substrate and enzyme concentrations. Initial rates are expressed as millimoles teriakiren degraded per minute, as shown in Table IV.
- single-point Ki determination of KI for a single inhibitor concentration
- multiple-point W determination of Ki for multiple concentrations
- the dissolution time is an important factor for the performance of the protecting agents of this invention.
- the reported dissolution time is the time required for a 0.5 mg/ml slurry of the test solid to dissolve completely in a 0.1 M citric acid/0.2M disodium phosphate pH 7 buffer at room temperature rotating end over end at 8 rpm. Visual inspection was used to determine the endpoint for complete dissolution.
- the % protein was determined for various materials which are protecting agents.
- the concentrations of carbon, hydrogen, and nitrogen in the sample were determined using a Perkin-Elmer 2400 C, H, and N Elemental Analyzer. Approximately two mg of sample was accurately weighed and placed into the analyzer. The % nitrogen in the sample was multiplied by 6.25 to give the estimate of % protein.
- EXAMPLE 12 The ability of a variety of commercial and processed protein fractions to reduce the degradation of terlakiren by chymotrypsin was determined. Soy flour was from Sigma Chem. Co.; almond flour and peanut flour were from Pert Labs; wheat gluten was from Total Foods Corp.
- Soy flour from Sigma Chemical Co. is unroasted, and thus contains active Bowman-Birk trypsin/chymotrypsin inhibitor, which has an 8000 MW.
- Soy protein (#PP620) from Protein Technologies, Inc. is a heat-treated preparation, in which the Bowman-Birk trypsin/chymotrypsin inhibitor has been inactivated. Percent inhibition was determined as described in Example 8. Table V demonstrates that the tested protecting agent fractions reduce the chymotrypsin- catalyzed degradation of terlakiren, a chymotrypsin-sensitive renin inhibitor.
- EXAMPLE 13 For a variety of commercially available protein materials and for processed protein fractions, the % protein, dissolution time, and Ki were determined (as described in Bcamples 11, 10, and 9, respectively). These data are presented in Table VI. These data demonstrate that degradation-reducing fractions can be prepared which exhibit both a low Ki and a short dissolution time. Soy flour from Sigma Chemical Co. is unroasted, and thus contains active
- BBMV Cholecystokinin-8
- Rat jejunal brush border membrane vesicles were prepared according the method of Kessler et al (Biochem. Biophys. Acta 506 (1978) 136).
- BBMV 25 microgm protein
- CCK-8 10 micromolar
- protecting agents 0.15 mg/ml
- Samples were withdrawn at 1 and 3 minutes, were quenched by acetonitrile in an ice bath, and were assayed for undegraded CCK-8 using a high performance liquid chromatography assay.
- the pentapeptide enkephalin analogue D-Ala-D-Leu-enkephalin (YdAGFdL) (1.0 mg) was directly ileally dosed to chronically ileally fistulated rats.
- Radiolabeled YdAGFdL (1.12 microgram) was dosed with each of four protecting agents: (1) decanted soy flour, ⁇ 1 K MW; (2) decanted and ultrafiltered soy flour, 1 K-30K MW; (3) pepsin-treated (FMC ACTl-MOD) soy flour, ⁇ 30K MW; (4) the potent aminopeptidase inhibitor amastatin (positive control).
- Table VIII presents absolute bioavailabilities from various treatments.
- the pepsin-treated (FMC ACTl-MOD) soy flour (MW ⁇ 30K) was particularly effective as a protecting agent against intestinal aminopeptidases, as evidenced by an almost 11 -fold increase in % absorbed.
- the potent aminopeptidase inhibitor amastatin was also effective, demonstrating that the bioavailability of YdAGFdL is partially limited by degradation by intestinal aminopeptidases. Unroasted soy flour offered no protection to degradation by intestinal aminopeptidases.
- Soy flour (Sigma Chem. Co.) (36 gm) was suspended in 1080 ml water and 120 ml of a 0.1% (w/v) solution of Thimerosol. The suspension was mixed at room temperature for 24 hr, and centrifuged for 1 hr at 3600 ⁇ m. The supernatant was ultrafiltered using a 30K MWCO membrane. The >30K MW fraction was collected, concentrated, and adjusted to pH 2 with 0.5 N HCI. This >30K MW fraction was fed into an ACTl-MOD spiral reactor module loaded with 8 gm immobilized pepsin (FMC Co ⁇ ., Pinebrook, NJ). The outflow of he enzyme reactor was ultrafiltered using a 30K MWCO membrane.
- % inhibition 100% x [1 - (S ⁇ SJ], where, S ⁇ is the rate of change of absorbance with time in the presence of inhibitor and S 0 is the rate of change of absorbance with time in the absence of inhibitor.
- Percent inhibition was determined using filtered soy flour, 30K - 100K fraction.
- Enzymatic treatment is carried out with a single proteolytic enzyme, or with various combinations of proteolytic enzymes, acting concurrently or sequentially.
- a variety of proteolytic enzymes are used, including but not limited to trypsin, chymotrypsin, elastase, carboxypeptidase, aminopeptidase, pepsin, and collagenase.
- Example 1-8 The fractionation and enzymatic treatments of Examples 1-8 are applied to a wide variety of proteinaceous materials of animal or vegetable origin.
- proteinaceous starting materials include but are not limited to soy flour, soy protein, wheat gluten, almond powder, peanut powder, casein, and fish protein.
- EXAMPLE 19 The ability of a protecting agent to dissolve quickly and to begin acting immediately upon being released in vivo is an important factor for the performance of the degration-reducing agents of this invention.
- soy flour and of 30K-100K soy flour, decanted and ultrafiltered, to reduce the degradation of Terlakiren in a dynamic environment were compared in vitro.
- the in vitro methodology of Example 8 was followed except the test solution contained only enzyme and Teriakiren in buffer.
- Test inhibitor (soy flour or 30K-100K soy flour, decanted and ultrafiltered) was added at a concentration of 0.01 mg/ml without additional mixing, while solution was shaking at speed 5 in a water bath (American Scientific, model # YB531) at 37° C. Sampling was at 19 seconds, 1 minute, then 2 minute intervals until 11 minutes had elapsed, then at 5 minute intervals for a total of 46 minutes. Quenching, HPLC analysis and data analysis were as in Example 8.
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Gastroenterology & Hepatology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Medicinal Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Epidemiology (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Immunology (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Biophysics (AREA)
- Zoology (AREA)
- Botany (AREA)
- Inorganic Chemistry (AREA)
- Endocrinology (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
Description
Claims
Priority Applications (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP5510892A JPH06510796A (en) | 1991-12-18 | 1992-11-09 | Soy protein or hydrolyzate in pharmaceutical compositions for protecting bioactive peptides from enzymatic inactivation |
EP92924173A EP0617626A1 (en) | 1991-12-18 | 1992-11-09 | Soybean protein or hydrolyzates in pharmaceutical compositions to protect bioactive peptides from enzymatic inactivation |
US08/244,715 US6153592A (en) | 1992-11-09 | 1992-11-09 | Enhancing the bioavailability of proteolytically labile therapeutic agents |
FI942938A FI942938A (en) | 1991-12-18 | 1994-06-17 | Soy protein or hydrolysates in pharmaceutical compositions for protection of bioactive peptides against enzymatic inactivation |
NO942323A NO942323L (en) | 1991-12-18 | 1994-06-17 | Soy protein or hydrolysates in pharmaceutical compositions for the protection of bioactive peptides against enzymatic inactivation |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US81059391A | 1991-12-18 | 1991-12-18 | |
US810,593 | 1991-12-18 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO1993011799A1 true WO1993011799A1 (en) | 1993-06-24 |
Family
ID=25204183
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/US1992/009336 WO1993011799A1 (en) | 1991-12-18 | 1992-11-09 | Soybean protein or hydrolyzates in pharmaceutical compositions to protect bioactive peptides from enzymatic inactivation |
Country Status (12)
Country | Link |
---|---|
EP (1) | EP0617626A1 (en) |
JP (1) | JPH06510796A (en) |
AU (1) | AU3058392A (en) |
CA (1) | CA2125400A1 (en) |
FI (1) | FI942938A (en) |
HU (1) | HUT69785A (en) |
IL (1) | IL104048A0 (en) |
MX (1) | MX9207407A (en) |
NO (1) | NO942323L (en) |
PT (1) | PT101135A (en) |
WO (1) | WO1993011799A1 (en) |
ZA (1) | ZA929761B (en) |
Cited By (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1994007472A1 (en) * | 1992-10-02 | 1994-04-14 | Pfizer Inc. | Pharmaceutical compositions containing nonionic surfactants |
WO1995006463A1 (en) * | 1993-09-01 | 1995-03-09 | Pfizer Inc. | Pharmaceutical compositions containing anionic surfactants |
WO1997007814A1 (en) * | 1995-08-22 | 1997-03-06 | Bionebraska, Inc. | Method and composition for enhanced parenteral nutrition |
WO2001054514A1 (en) * | 2000-01-27 | 2001-08-02 | Aqua Solution Inc. | Composition for intestinal delivery |
GB2367821A (en) * | 2000-06-30 | 2002-04-17 | Nat Biolog Standards Board | Stabilisation of cytokine solutions by casein |
US6852690B1 (en) | 1995-08-22 | 2005-02-08 | Amylin Pharmaceuticals, Inc. | Method and composition for enhanced parenteral nutrition |
US20120004157A1 (en) * | 2007-06-08 | 2012-01-05 | Bergen Teknologioverforing As | Hydroxyproline compositions and uses thereof |
Citations (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0090886A2 (en) * | 1982-04-06 | 1983-10-12 | American Cyanamid Company | Stabilization of influenza virus vaccine |
EP0127535A2 (en) * | 1983-05-23 | 1984-12-05 | Hadassah Medical Organization | Pharmaceutical compositions containing insulin |
EP0266950A2 (en) * | 1986-10-31 | 1988-05-11 | Pfizer Inc. | Nor-statine and nor-cyclostatine polypeptides |
US4774089A (en) * | 1985-06-14 | 1988-09-27 | Albion International, Inc. | Stimulation of gonadotropic hormones with mineral mixtures containing amino acid chelates |
EP0327756A2 (en) * | 1987-11-13 | 1989-08-16 | Smithkline Beecham Farmaceutici S.p.A. | Pharmaceutical compositions comprising a calcitonin and a glycyrrhizinate as absorption enhancer |
EP0341661A2 (en) * | 1988-05-11 | 1989-11-15 | The Du Pont Merck Pharmaceutical Company | Peptide-drug compositions containing alpha-aminoboronic acid derivatives |
WO1990003164A2 (en) * | 1988-09-29 | 1990-04-05 | Patralan Limited | Pharmaceutical formulations |
EP0371195A1 (en) * | 1988-07-12 | 1990-06-06 | BERLIN-CHEMIE Aktiengesellschaft | Process for producing preparations containing acitve peptides suited for rectal and vaginal application |
WO1991006292A1 (en) * | 1989-11-02 | 1991-05-16 | Danochemo A/S | Process of preparing a water dispersible hydrophobic or aerophilic solid |
-
1992
- 1992-11-09 CA CA002125400A patent/CA2125400A1/en not_active Abandoned
- 1992-11-09 EP EP92924173A patent/EP0617626A1/en not_active Withdrawn
- 1992-11-09 AU AU30583/92A patent/AU3058392A/en not_active Abandoned
- 1992-11-09 HU HU9401824A patent/HUT69785A/en unknown
- 1992-11-09 WO PCT/US1992/009336 patent/WO1993011799A1/en not_active Application Discontinuation
- 1992-11-09 JP JP5510892A patent/JPH06510796A/en active Pending
- 1992-12-10 IL IL104048A patent/IL104048A0/en unknown
- 1992-12-16 PT PT101135A patent/PT101135A/en not_active Application Discontinuation
- 1992-12-17 ZA ZA929761A patent/ZA929761B/en unknown
- 1992-12-17 MX MX9207407A patent/MX9207407A/en unknown
-
1994
- 1994-06-17 FI FI942938A patent/FI942938A/en not_active Application Discontinuation
- 1994-06-17 NO NO942323A patent/NO942323L/en unknown
Patent Citations (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0090886A2 (en) * | 1982-04-06 | 1983-10-12 | American Cyanamid Company | Stabilization of influenza virus vaccine |
EP0127535A2 (en) * | 1983-05-23 | 1984-12-05 | Hadassah Medical Organization | Pharmaceutical compositions containing insulin |
US4774089A (en) * | 1985-06-14 | 1988-09-27 | Albion International, Inc. | Stimulation of gonadotropic hormones with mineral mixtures containing amino acid chelates |
EP0266950A2 (en) * | 1986-10-31 | 1988-05-11 | Pfizer Inc. | Nor-statine and nor-cyclostatine polypeptides |
EP0327756A2 (en) * | 1987-11-13 | 1989-08-16 | Smithkline Beecham Farmaceutici S.p.A. | Pharmaceutical compositions comprising a calcitonin and a glycyrrhizinate as absorption enhancer |
EP0341661A2 (en) * | 1988-05-11 | 1989-11-15 | The Du Pont Merck Pharmaceutical Company | Peptide-drug compositions containing alpha-aminoboronic acid derivatives |
EP0371195A1 (en) * | 1988-07-12 | 1990-06-06 | BERLIN-CHEMIE Aktiengesellschaft | Process for producing preparations containing acitve peptides suited for rectal and vaginal application |
WO1990003164A2 (en) * | 1988-09-29 | 1990-04-05 | Patralan Limited | Pharmaceutical formulations |
WO1991006292A1 (en) * | 1989-11-02 | 1991-05-16 | Danochemo A/S | Process of preparing a water dispersible hydrophobic or aerophilic solid |
Non-Patent Citations (1)
Title |
---|
JOURNAL OF CONTROLLED RELEASE vol. 13, no. 2+3, August 1990, NEW YORK pages 213 - 223 VINCENT H.L.LEE 'Protease inhibitors and penetration enhancers as approaches to modify peptide absortion' cited in the application * |
Cited By (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1994007472A1 (en) * | 1992-10-02 | 1994-04-14 | Pfizer Inc. | Pharmaceutical compositions containing nonionic surfactants |
WO1995006463A1 (en) * | 1993-09-01 | 1995-03-09 | Pfizer Inc. | Pharmaceutical compositions containing anionic surfactants |
WO1997007814A1 (en) * | 1995-08-22 | 1997-03-06 | Bionebraska, Inc. | Method and composition for enhanced parenteral nutrition |
US6852690B1 (en) | 1995-08-22 | 2005-02-08 | Amylin Pharmaceuticals, Inc. | Method and composition for enhanced parenteral nutrition |
US7569540B2 (en) | 1995-08-22 | 2009-08-04 | Amylin Pharmaceuticals, Inc. | Method for enhanced parenteral nutrition |
WO2001054514A1 (en) * | 2000-01-27 | 2001-08-02 | Aqua Solution Inc. | Composition for intestinal delivery |
GB2367821A (en) * | 2000-06-30 | 2002-04-17 | Nat Biolog Standards Board | Stabilisation of cytokine solutions by casein |
GB2367821B (en) * | 2000-06-30 | 2002-11-20 | Nat Biolog Standards Board | Stabilisation of cytokines by casein |
US20120004157A1 (en) * | 2007-06-08 | 2012-01-05 | Bergen Teknologioverforing As | Hydroxyproline compositions and uses thereof |
Also Published As
Publication number | Publication date |
---|---|
MX9207407A (en) | 1993-06-01 |
IL104048A0 (en) | 1993-05-13 |
NO942323D0 (en) | 1994-06-17 |
ZA929761B (en) | 1994-06-17 |
HU9401824D0 (en) | 1994-09-28 |
CA2125400A1 (en) | 1993-06-24 |
FI942938A0 (en) | 1994-06-17 |
HUT69785A (en) | 1995-09-28 |
EP0617626A1 (en) | 1994-10-05 |
PT101135A (en) | 1994-03-31 |
NO942323L (en) | 1994-06-17 |
JPH06510796A (en) | 1994-12-01 |
FI942938A (en) | 1994-06-17 |
AU3058392A (en) | 1993-07-19 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
KR0138658B1 (en) | Intravaginal delivery of biologically active polypeptides | |
RU2198677C2 (en) | Pharmaceutical composition for oral delivery of physiologically active peptide agent and method of enhancement of its availability | |
JP5100334B2 (en) | Encapsulated clear liquid for drug delivery | |
EP0134479B1 (en) | Modified proteinase inhibitors, process for their preparation and pharmaceutical compositions containing them | |
Sinha et al. | Oral colon-specific drug delivery of protein and peptide drugs | |
JP2010509308A (en) | Transdermal delivery system for peptides and related compounds | |
JP2805194B2 (en) | Peptide for increasing blood triglyceride concentration and blood triglyceride concentration increase inhibitor containing the peptide as an active ingredient | |
GB2251435A (en) | Opioid peptides derived from wheat proteins | |
EP0177605B1 (en) | An enzyme composition acting as a digestion promotor on various levels in the alimentary tract, and a method for facilitating digestion | |
Bai et al. | Targeting of peptide and protein drugs to specific sites in the oral route | |
WO1993011799A1 (en) | Soybean protein or hydrolyzates in pharmaceutical compositions to protect bioactive peptides from enzymatic inactivation | |
JP2764276B2 (en) | Functional novel peptides and their use | |
US6153592A (en) | Enhancing the bioavailability of proteolytically labile therapeutic agents | |
EP1874329B1 (en) | Blood pressure lowering protein hydrolysates | |
Hosny et al. | Promotion of oral insulin absorption in diabetic rabbits using pH-dependent coated capsules containing sodium cholate | |
CA2165787A1 (en) | Medical use of stem bromelain protease | |
JP2004010569A (en) | Arginine-containing peptide having cholecystokinin secretion promoting activity and food containing the same | |
LEE | Peptidase activities in absorptive mucosae | |
JP2004010569A5 (en) | ||
JPH05148154A (en) | Pharmaceutical containing physiologically active polypeptide | |
JP2952830B2 (en) | Antihypertensive | |
Bai et al. | Comparison of site-dependent degradation of peptide drugs within the gut of rats and rabbits | |
AU2016213759B2 (en) | Transdermal delivery systems of peptides and related compounds | |
JP2002511867A (en) | Use of bromelain in the manufacture of a medicament for enhancing intestinal permeability | |
AU2014203176B2 (en) | Transdermal delivery systems of peptides and related compounds |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AK | Designated states |
Kind code of ref document: A1 Designated state(s): AU CA FI HU JP KR NO US |
|
AL | Designated countries for regional patents |
Kind code of ref document: A1 Designated state(s): AT BE CH DE DK ES FR GB GR IE IT LU MC NL SE |
|
DFPE | Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101) | ||
WWE | Wipo information: entry into national phase |
Ref document number: 2125400 Country of ref document: CA |
|
WWE | Wipo information: entry into national phase |
Ref document number: 1992924173 Country of ref document: EP |
|
WWE | Wipo information: entry into national phase |
Ref document number: 942938 Country of ref document: FI |
|
WWE | Wipo information: entry into national phase |
Ref document number: 08244715 Country of ref document: US |
|
WWP | Wipo information: published in national office |
Ref document number: 1992924173 Country of ref document: EP |
|
WWW | Wipo information: withdrawn in national office |
Ref document number: 1992924173 Country of ref document: EP |