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WO1993021225A1 - Peptide synthetique, tensioactif pulmonaire le renfermant, et remede contre le syndrome de souffrance respiratoire - Google Patents

Peptide synthetique, tensioactif pulmonaire le renfermant, et remede contre le syndrome de souffrance respiratoire Download PDF

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Publication number
WO1993021225A1
WO1993021225A1 PCT/JP1993/000492 JP9300492W WO9321225A1 WO 1993021225 A1 WO1993021225 A1 WO 1993021225A1 JP 9300492 W JP9300492 W JP 9300492W WO 9321225 A1 WO9321225 A1 WO 9321225A1
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Prior art keywords
val
leu
peptide
lie
surfactant
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PCT/JP1993/000492
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English (en)
Japanese (ja)
Inventor
Tsunetomo Takei
Toshimitsu Aiba
Kaoru Sakai
Tetsuro Fujiwara
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Mitsubishi Tanabe Pharma Corp
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Tokyo Tanabe Co Ltd
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Priority to JP51818893A priority Critical patent/JP3376582B2/ja
Publication of WO1993021225A1 publication Critical patent/WO1993021225A1/fr
Anticipated expiration legal-status Critical
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/785—Alveolar surfactant peptides; Pulmonary surfactant peptides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • 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 synthetic peptides. More specifically, the present invention relates to a synthetic peptide having a strong surface activity by being combined with a lipid mixture, a pulmonary surfactant comprising the synthetic peptide and a lipid mixture, and a therapeutic agent for respiratory distress syndrome containing the pulmonary surfactant as an active ingredient. . Background technology,
  • Respiratory distress syndrome is a disease that causes severe respiratory distress as a result of the collapse of the alveoli due to a lack of pulmonary surfactant.
  • Pulmonary surfactants to be supplemented include substances consisting of phospholipids, neutral lipids, total cholesterol and carbohydrates and a small amount of proteins present in lung tissue of mammals (Japanese Patent Publication No. 61-92925), In addition to the components, a substance containing fatty acids (hereinafter referred to as “S-TA”: Japanese Patent Publication No. 61-92924), a substance separated from pig lung lavage fluid and added with Ca (Japanese Journal of the Japanese Society of Surface Medicine, Vol. 12, No. 1, p.
  • Some of the present inventors previously separated a lipoprotein from an animal-derived lung surfactant, and added the lipoprotein to a lipid mixture because the lipoprotein is an essential component for exhibiting lung surface activity.
  • the surfactant exerts an excellent surface tension lowering effect, shortens the surface wavefront diffusion effect of the surfactant, and exerts a low surface tension on the flat mouth, thereby securing sufficient alveolar cavity volume, etc., resulting in respiratory distress syndrome. It has been discovered that it can be used to treat (Japanese Patent Publication No. 3-788371).
  • SP-C derived from human lung is an apoprotein with extremely strong hydrophobicity, which is composed of 35 amino acid residues and whose N-terminal amino acid is phenylalanine and rich in hydrophobic amino acids such as amino acids.
  • SP-C isolated from the lungs of pigs, pigs, rats, etc. also consists of 34 to 35 amino acids, and although the amino acid sequence at the N-terminal varies depending on the animal species, it is not Are extremely high in homology.
  • Val Val Val Val Val Leu lie Val Val Val lie Val Gly Ala Leu Leu
  • SP-B or SP-C promotes adsorption and diffusion of lung surfactant to the gas-liquid interface. To improve the surface activity of pulmonary surfatatantes.
  • Therapeutic agent for respiratory distress syndrome which contains a pulmonary surfactant consisting of SP-C and a lipid mixture as an active ingredient, is extremely effective, but SP-C is extremely hydrophobic and difficult to isolate and purify. Due to the fact that it is contained in living organisms in extremely small amounts, it has not been put to practical use.
  • Japanese Patent Application Laid-Open No. 3-52095 states that a mixture of a synthetic peptide containing the SP-C partial structure shown below and a lipid is effective for the treatment of respiratory distress syndrome Have been.
  • the publication also describes a peptide having 32 amino acid residues having the following sequence as a minimum unit exhibiting high surface activity.
  • the pulmonary surfactant preparation is often provided as a dry powder preparation which is converted into a physiological saline suspension wave at the time of use from the viewpoint of quality conservation.
  • S-35 pulmonary surfactant preparations in which a synthetic peptide having the amino acid sequence of SP-C is mixed with a lipid mixture composed of choline phosphoglyceride, acidic phospholipid, and fatty acids, are: Since the cysteine residue present in the peptide forms a disulfide bond, the dispersibility in physiological saline is extremely poor due to factors such as high peptide agglutinability and strong hydrophobicity of the lung surfactant itself. It was difficult to make the suspension uniform enough to be used as such.
  • a method for improving the suspendability of a pulmonary surfactant preparation a method of adding a suspending agent such as mannitol (Japanese Patent Application Laid-Open No. Sho 60-34905) and a method in which the temporary freezing temperature during drying is reduced to 1: 1.
  • a freezing method performed at temperatures of up to 11 ° C. has been proposed (Japanese Patent Application Laid-Open No. Sho 63-107718), the operation is complicated, and it is hoped that a more convenient method for producing a drug product will be developed. Disclosure of the Invention
  • the inventors of the present invention have conducted intensive studies on a synthetic peptide having a strong surface activity by being combined with a lipid mixture, and as a result, a peptide represented by the following specific sequence (hereinafter referred to as “the synthetic peptide of the present invention”)
  • the pulmonary surfatatant containing) as an active ingredient was produced by a conventional freeze-drying method performed at 120 ° C or lower without the addition of a suspending agent, even when S-35, corin phosphoglycerol was used.
  • Only lipid mixtures consisting of acidic phospholipids and fatty acids It has better uniform suspension than S-TA, and has the same strong surface activity as S-35 or S-TA. Knowing this, they completed the present invention.
  • Xaa is absent or represents Cys, Ser or Ala
  • Xbb represents Cys, Ser or Ala
  • Xcc represents His or Asn
  • Xdd represents Leu or Ile
  • Xee is Val or l ie
  • Xff represents I le, Leu or Val
  • Xgg does not exist or represents Leu.
  • the present invention is a synthetic peptide which is easy to isolate and purify due to the low frequency of production of immature peptides during production, and can be produced in large quantities in a short time.
  • a synthetic peptide having good suspension properties and strong surface activity by being blended, a lung surfactant comprising a mixture of the synthetic peptide and lipid, and a treatment for respiratory distress syndrome containing the lung surfactant as an effective component An agent is provided.
  • the synthetic peptide of the present invention can be produced by a chemical or genetic engineering technique, but a chemical production method is preferred in terms of isolation and purification.
  • Examples of chemical production methods include “Peptide synthesis (by Nobuo Izumiya, Maruzen Co., Ltd., 1979)”, “Biochemistry—Lecture, Volume 1, Protein Chemistry IV, Chemical Modification and Peptides” Synthesis-(Shunpei Sakakibara, Tokyo Chemical Doujin Co., Ltd., 1973) "," Semi-chemical Chemistry Laboratory, Vol. 2, Protein Chemistry (below), Peptide Synthesis (Kimura Terutoshi, Tokyo Chemical) Dojin Co., Ltd., 1987), "Solid phase peptide synthesis, a practical approach;, Senoreton (E.
  • Azide method acid chloride method, acid anhydride method, mixed acid anhydride method, DCC method, active ester method ( ⁇ -ditrophenyl ester method, P-hydroxysuccinic acid imid ester method, etc.), carboimidazole It can be manufactured by a liquid phase synthesis method such as a redox method, a redox method, a DCC-activation method, or a solid synthesis method.
  • solid phase synthesis is preferred, and synthesis can be performed by an automatic synthesizer.
  • the automatic synthesizer include a 431A peptide synthesizer (trademark; manufactured by Applied Biosystems) or a peptide synthesizer model 9900E (trademark; manufactured by Beckman).
  • a pulmonary surfactant (hereinafter referred to as “surfactant of the present invention”) can be produced by blending choline phosphoglyceride, acidic phospholipid and fatty acids as a lipid mixture with the synthetic peptide of the present invention.
  • the compounding ratio is 0.1 to 5.0% (W / W) for synthetic peptide and 50.6 to 85.0 for choline phosphoglyceride, based on the weight ratio of these components to the total dry weight of the final product.
  • % (W / W) ⁇ It is appropriate to set acidic phospholipids to 4.5 to 37.6% (W / W) and fatty acids to 4.6 to 24.6% (WZW). It is.
  • Choline phosphodalicerides that can be used in the surfactants of the present invention include 1,2-dipalmitoyl glyceride— (3.)-Phosphocholine (also known as dipalmitoyl phosphatidyl choline) and 1,2-distearoyl glyceride.
  • corynephosphoglyceride may be an acyl group having 12 to 24 carbon atoms, preferably two saturated acyl groups.
  • 2-Diacylglycerose- (3) -A mixture of two or more phosphocholins, and a mixture of the mixture and the above-mentioned single product can also be used.
  • acidic phospholipids examples include 1,2-diacyl-sn-glycerol (3) monophosphoric acid (also known as L- ⁇ -phosphatidic acid), 1,2-diacyl-s ⁇ -glycerol— (3) -phospho-serine (Also known as phosphatidylserine), 1,2-diasyl-s ⁇ -glycerol— (3) —phospho-s ⁇ -glycerol (also known as phosphatidylglycerol) or 1,2-diasyl-s ⁇ -glycerol (3) —phospho (1) — L-1 my 0—Inositol (also known as phosphatidylinositol) is suitable.
  • the 1-position and the 2-position may be substituted with the same or different kinds of acyl groups, respectively.
  • the carbon number of the acyl group is preferably from 12 to 24.
  • Fatty acids include free fatty acids, alkali metal salts of fatty acids, and fatty acids. Suitable are & alkyl esters, fatty acid glycerin esters, fatty acid amides, or mixtures of two or more of these, and furthermore, fatty alcohols or aliphatic amines.
  • on fatty acids is meant to include the fatty alcohols and aliphatic amines referred to herein.
  • Myristic acid, palmitic acid or stearic acid is suitable as a free fatty acid. Lumitic acid !? Good.
  • sodium palmitate is used as the metal salt of fatty acid
  • ethyl palmitate is used as fatty acid alkyl ester
  • monopalmitin is used as fatty acid glycerin ester
  • palmitic acid amide is used as fatty acid amide.
  • Hexadecyl alcohol is preferred as the fatty alcohol, and hexadecylamine is preferred as the aliphatic amine.
  • choline phosphoglycerides, acidic phospholipids and fatty acids may be any of products isolated from animals and plants, semi-synthetic products or chemically synthesized products, and commercially available products thereof can be used.
  • the surfactant of the present invention is obtained by drying a mixed solution of the synthetic peptide solution of the present invention and the above-mentioned lipid mixture or compound solution under reduced pressure, suspending the obtained residue using a suitable suspending solvent, and then freeze-drying. It can be manufactured by a method.
  • the solvent used for preparing the synthetic peptide solution of the present invention include formic acid, trifluoroacetic acid (TFA), trifluoroethanol, dimethyl sulfoxide (DMS0), chloroform / methanol, and chloroform.
  • Examples of the solvent used for preparing the lipid mixture solution include chloroform, chloroform / methanol [2: 1 to 5: 1. (V / V)] —.
  • Examples of the suspending solvent include water or a mixed solution of water and ethanol [4: 1 to 20: 1 (V / V)], and a mixed solution of water and ethanol is preferable.
  • the suspension is carried out at 30 to 60 ° C., preferably 40 to 50 ° C., for 5 to 60 minutes, preferably for 15 to 30 minutes.
  • the surfactant of the present invention it is inevitable that a trace amount of water remains in the production method, but it is preferable to dry the surfactant until the remaining weight ratio becomes 5.0% (W / W) or less based on the total weight. If it is dried to such an extent, residual ethanol cannot be detected when a water-ethanol mixture is used.
  • the surfactant dry powder preparation of the present invention can be prepared in a variable-speed mixer or an ultrasonic generator at an appropriate physiological concentration of a monovalent or divalent metal salt, for example, 0.9% sodium chloride or 1.5%. It can be used by uniformly suspending and dispersing it using mM calcium chloride or a physiological buffer containing them.
  • a monovalent or divalent metal salt for example, 0.9% sodium chloride or 1.5%. It can be used by uniformly suspending and dispersing it using mM calcium chloride or a physiological buffer containing them.
  • the surface tension lowering effect was measured according to the method of Tanaka et al. (Journal of the Surface Science Society of Japan, Vol. 13, No. 2, No. 87, p. 87, 1982).
  • the surfactant of the present invention is dropped onto physiological saline (surface area: 54.0 cm 2 ) so that the surfactant of the present invention is 1.0 to 2.0 / g per cm 2 , and the surface area is reduced. 54.
  • the surface tension during compression and expansion in the range of 0 to 2 1.6 cm 2 over 2 to 5 minutes was measured at 37 ° C with a Wilhelmy surface tension measurement device (manufactured by Kyowa Interface Science Co., Ltd.). Measured continuously.
  • the surface tension reducing effect of the surfactant of the present invention is as follows.
  • the initial surface tension of saline at 37 ° C was 70.5 dyne
  • the arrival time refers to the time required for the surface tension to reach a constant value immediately after dropping of the sample, and the equilibrium surface tension refers to the value at that time.
  • the surfactant of the present invention formed a film on the gas-liquid surface in a short time of 3 to 65 seconds, and reduced the surface tension to 27.9 to 34.8 dyneZcm.
  • a suspension of physiological saline at 37 ° C containing 0.2 to 1.0 mg of the surfactant of the present invention per liter was prepared, and the suspension of the surfactant of the present invention to the saline gas-liquid surface was prepared.
  • the adsorption rate was measured according to the method of King et al. (American Journal of Physiology, Vol. 223, Vol. 7, pp. 15, 1972).
  • the suspension was poured into the bottom of a 5 cm diameter water tank containing physiological saline, and the mixture was stirred gently with a magnetic stirrer. The speed was determined.
  • the surfactant of the present invention reduced the surface tension to a range of 28.:! To 39.5 dyne cm after 30 to 120 seconds had elapsed after the stirring was stopped, and then showed a constant value.
  • the suspension test of the lung surfactant was performed according to the method disclosed in Japanese Patent Application Laid-Open No. Sho 63-107718.
  • the suspendability was determined based on the dispersion ratio at predetermined time intervals after the start of the suspension and the maximum dispersed particle diameter after 2 minutes from the start of the suspension.
  • the determination of the suspension state was carried out by two persons, 10 samples each time at each time, and no judgment was made as to whether or not the suspension was any small lump in the container, and the formulation was uniformly dispersed in physiological saline This was done by determining whether a white, slightly viscous suspension had formed.
  • the dispersion ratio was calculated as a percentage of the total number of samples (10 tubes) in which each individual completed the suspension at each time, and the average value was shown by the two individuals.
  • the maximum dispersed particle size was determined by dispensing 6 mg of lung surfactant into a 20 ml vial of each sample, injecting 2 ml of physiological saline, and shaking continuously for 2 minutes under the same shaking conditions as described above. The largest particle was found by using a microscope and its diameter was determined by measuring with a vernier caliper.
  • surfactants of the present invention were suspended within 2 minutes in most cases, and had a maximum particle diameter of 0.9 mm or less, and showed good suspendability.
  • the acute toxicity of the surfactant of the present invention was tested using 5-week-old male ICR mice and Dister rats. Oral LD 5 in mice. And intraperitoneal LD 5 . Is 2.5-10.0 / 3 ⁇ 4: and 1.5-5.O gZkg, and those on the rat are 1.5-5.0 11 and 1.5-2.5 g Z kg.
  • the surfactant of the present invention was intraperitoneally administered to mature Wistar rats for 3 months 0 to 600 mgZkg for 1 month, but no change in body weight or abnormality in macroscopic and histological observation of major organs was observed. .
  • a rabbit immature fetus with a gestation period of 27 days produces little pulmonary surfactant and is in a pulmonary surfactant-deficient state, and is therefore a model animal for neonatal respiratory distress syndrome.
  • lung volume the alveolar cavity volume (hereinafter, referred to as lung volume) was measured at 37 ° C under increasing and decreasing airway pressure.
  • the measurement was continuously performed 5 minutes after the surfactant of the present invention was intratracheally administered using a water manometer connected to the trachea by incising the neck of the fetus.
  • 2-channel independent drive cylinder with tracheal pressure connected to the trachea The pressure was increased to 30 cm water pressure using a dipump No. 940 (manufactured by Harvard, USA) to expand the alveoli.
  • the airway pressure was reduced to 0 cm water pressure, the alveoli were contracted, and the lung volume at each water pressure was measured. Lung volume was expressed in milliliters per kilogram of body weight (m1Zkg).
  • the administration of the surfactant of the present invention has a concentration of 1.0 to 6.0% (W /
  • V was performed by a method of injecting 0.05 to 0.5 ml of a physiological saline suspension prepared directly into the airway.
  • physiological saline was injected instead of the surfactant suspension of the present invention.
  • the lung capacity (5 cm water pressure) of the immature rabbit at gestational age 27 was 1-5 m 1 / kg, and the alveoli were hardly expanded.
  • gestational age 30 fetuses with normal levels of pulmonary surfactant have a lung volume (5 cm water pressure) of 39-53 m 1 / kg and alveolar dilation. Shows that it is possible to breathe normally.
  • the lung volume (5 cm water pressure) of the immature fetus was 15 to 25 m 1 / kg and the alveolar expansion was insufficient.
  • the lung capacity (5 cm water pressure) of the immature fetus to which the surfactant of the present invention was administered was 35 to 53 m 1 Zkg, indicating that the surfactant of the present invention improved the lung capacity of the immature fetus to a normal level.
  • the present synthetic peptide has an action of strongly activating the surface activity of the lipid mixture, and the present surfactant comprising the present synthetic peptide and the lipid mixture exhibits surface activity, suspendability and Based on its pharmacological properties, it is an effective remedy for respiratory distress syndrome.
  • the remedy for respiratory distress syndrome provided by the present invention is a single dose of 50 to 100 mg for children and 500 to 500 mg for adults.
  • This dose is suspended in water, physiological saline, or a physiologically acceptable buffer, and adjusted to a concentration of 1.0 to 10% by weight (WZV).
  • WZV 1.0 to 10% by weight
  • Use by injecting or spraying 1 to 10 times into the respiratory tract 48 hours immediately after. In addition, they can be inhaled directly as powder without being suspended. Dosage, use and frequency may be adjusted as appropriate for the patient's condition and combination therapy.
  • the therapeutic agent of the present invention may contain, if necessary, pharmaceutical additives such as stabilizers, preservatives, isotonic agents, buffers, suspending agents, or pharmaceuticals such as bronchodilators, antiallergic agents, anticancer agents, and antibacterial agents. It can be contained.
  • pharmaceutical additives such as stabilizers, preservatives, isotonic agents, buffers, suspending agents, or pharmaceuticals such as bronchodilators, antiallergic agents, anticancer agents, and antibacterial agents. It can be contained.
  • the dosage form is suitably a liquid preparation or a powder preparation to be used in suspension at the time of use.
  • the therapeutic agent of the present invention is filled in a sealed container such as a vial or ampoule and stored as a sterile preparation.
  • peptide A The peptide described in SEQ ID NO: 1 (hereinafter, referred to as “peptide A”) was converted to “Solid phase peptide synthesis” by E. Atherton and RC Sheppard. a practical approach) ”p. 25-189, 1989, Oxford University Press, Oxford] and KenicM, Akagi et al. [Chem. Pharm. Bull., 37 (10), p. 9 89)], the solid phase was synthesized using a multi-peptide solid-phase synthesis system “Koksan” (trade name; manufactured by Kokusaikagaku Co., Ltd.).
  • N- ⁇ -9-fluorenylmethyloxycarbo-2-leucine (Fmoc-Leu) was added to [4-1 (hydroxymethyl) Phenoxymethyl-copoly (styrene 1% divinylbenzene)] N- «-9-Fluorenylmethyloxycarbocarbone-to-resin bonded to resin-0-Resin (Fmoc-Leu-0- Resin) 0.2 g of 0.5 mm was used.
  • N-dimethylformamide (DMF) N-dimethylformamide
  • the resin was washed four times with DMF. A 20% piperidine-DMF solution was added and shaken to perform deprotection.
  • the amino acid was sequentially extended in the N-terminal direction on the resin to synthesize a peptide 0-resin in which the N-terminal and the functional group were completely protected.
  • the condensation reaction during the introduction of Arg, Lys, His. Pro, and Cys was performed twice in 120 minutes.
  • the amino group at the N-terminus of all amino acids was protected with an Fmoc group, and the functional side chains were protected with the following groups.
  • CZM formaldehyde-methanol
  • the presence of the peptide in the eluate ⁇ was monitored by 245 nm (spectrophotometer; Model 870-UV of JASCO Corporation) and a differential refractometer (Shimadzu Corporation; Model R ID-6A).
  • Boc-Leu t-butyloxycarboxy-l-isocyanate
  • B 0 c—L eu—PAM resin (0.70 mol / g, 0.35 g) is used as a reaction vessel in a peptide synthesizer (Model 990E, manufactured by Beckman).
  • a peptide synthesizer Model 990E, manufactured by Beckman.
  • Moved to The protected amino acid was extended in the N-terminal direction on the resin by a preformed symmetric anhydride method to synthesize a completely protected peptide 10-resin.
  • the completely protected peptide 0—resin (155 mg) was swollen in methylene chloride for 5 minutes.
  • the N—a—B0c protecting group was deprotected using TFA containing 1% (v / v) indole and 0.1% (v / v) ethanedithiol.
  • the deprotected peptide-0-resin was then treated with p-cresol (1 ml), p-thiocresol (0.2 g) and DMSO (Lm1) in anhydrous hydrogen fluoride (HF) ( 1 1 m 1) at 0 ° C for 60 minutes Upon treatment, the peptide was excised from the resin.
  • HF hydrous hydrogen fluoride
  • HF and DMS 0 were distilled off at 0 ° C. under vacuum.
  • the cut out peptide and resin were washed three times with 15 ml of cold getyl ether, and then the free peptide was extracted by washing three times with 10 ml of cold TFA washing solution.
  • the extract was filtered immediately and added to ice-cold water (12 Om1-150 ml) to precipitate crude peptide B.
  • the crude peptide B was centrifuged at 1,000 X g, 0 ° C for 30 minutes and collected as a precipitate.
  • the precipitate was washed with getyl ether (15 ml). This washing step was further repeated using getyl ether, ethyl acetate and distilled water to obtain 83 mg of peptide B.
  • the obtained peptide B was dissolved in a 50% aqueous solution of DMS and purified by high-performance liquid chromatography using a Bondasphere and C8-300 column.
  • a 50% aqueous solution of acetonitrile containing 1% TFA was used for elution for 5 minutes.
  • the eluate was then eluted with a linear concentration gradient of 80% acetonitrile aqueous solution containing 0.1% TFA for 30 minutes.
  • the presence of the peptide in the eluate was determined at 245 nm (spectrophotometer; JASCO Corporation). It was monitored with a model company model 870-UV) and a differential refractometer (Shimadzu Corporation Model RID-6A).
  • the peptide of SEQ ID NO: 3 (peptide C) was prepared in the same manner as in (Example 1).
  • the peptide of SEQ ID NO: 5 (peptide E) was prepared in the same manner as in (Example 1).
  • the peptide of SEQ ID NO: 6 (peptide F) was prepared in the same manner as in (Example 1).
  • the peptide of SEQ ID NO: 7 (peptide G) was prepared in the same manner as in (Example 1).
  • the peptide of SEQ ID NO: 8 (peptide H) was prepared in the same manner as in (Example 1).
  • the peptide of SEQ ID NO: 9 (peptide I) was prepared in the same manner as in (Example 1).
  • the peptide of SEQ ID NO: 10 (peptide J) was prepared in the same manner as in (Example 1).
  • the synthetic peptide of the present invention was prepared using 1,2N hydrochloric acid ZTFA [2: 1 (V / V)] containing 5% (v / v) funinol under vacuum at 150 ° C for 2, 4, 6, 1 After acid hydrolysis for 2, 24, 48 and 72 hours to remove the acid, the hydrolyzed products were analyzed by Shimadzu Amino Acid Automatic Analysis System (LC-19A). In the hydrolysis for 2 to 72 hours, the amino acid value showing higher recovery was adopted, and the amino acid composition value was calculated.
  • the molecular weight of the synthetic peptide of the present invention was measured by the fast atom bombardment method (FABMS).
  • FABMS fast atom bombardment method
  • a JMS-S102A type manufactured by JEOL Ltd.
  • a cesium gun (10 KeV) was used as an ion source.
  • the surfactant of the present invention is prepared by using the synthetic peptide and 1,2-dipalmitoylglycerol (3) -phosphocholine, 1,2-diasyl-sn-glycerol- (3) -phospho-sn-glycerol and palmitic acid as lipid components.
  • the three components of the acid were prepared by mixing.
  • 1,2-Dipalmitoyl glycerol— (3) Phosphocholine 204 mg, 1,2-Diacyl-s ⁇ -glycerol (3) —Phosphos ⁇ -glycerol (Chain 14 to 24 carbon atoms: Sigma) 6 3.
  • Om1 and 2.8 mg of peptide B was added to 3 ml of TFAO. Dissolved. These solutions were mixed and evaporated to dryness under reduced pressure.
  • the obtained residue was suspended in 100 ml of a water-ethanol mixture [9: 1 (V / V)] at 45 ° C for 20 minutes.
  • the suspension was frozen at ⁇ 60 ° C. and dried at a vacuum of 60 to 110 ° Hg for 40 hours to obtain 301.9 mg of a white powdered surfactant.
  • a surfactant was produced in the same manner as in (Example 12) except that peptide C was used instead of peptide B.
  • Surfactants were produced in the same manner as in (Example 12) except that peptide D was used instead of peptide B.
  • a surfactant was produced in the same manner as in (Example 12) except that peptide E was used instead of peptide B.
  • Surfactants were produced in the same manner as in (Example 12) except that peptide F was used instead of peptide B.
  • 1,2-dipalmitoyl glycerol— (3) phosphocholin 210 mg, 1,2-diacyl-s ⁇ -glycerol (3) —phospho-s ⁇ —glyceric ester (carbon number of the acyl group: 14 to 2 90 mg of Omg and 33.Omg of palmitic acid were dissolved in a mixture of form-methanol [3: 1 (V / V)] 40 Om1 and 3.Omg of peptide D was dissolved. Dissolved in 5 ml of TFAO. These solutions were mixed and evaporated to dryness under reduced pressure. The obtained residue was suspended in 120 ml of a water-ethanol mixture [9: 1 (V / V)] at 50 ° C for 15 minutes. This suspension was frozen in one 6 O e C was dried 2 8 hours at a vacuum degree 5 0 ⁇ 1 0 0 H g, was 3 3 7. 9 mg of Safakutan bets white powder.
  • a surfactant was produced in the same manner as in (Example 17) except that peptide H was used instead of peptide G.
  • Surfactant was produced in the same manner as in (Example 17) except that peptide J was used instead of peptide G.

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  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)

Abstract

Sont décrits un peptide représenté par la séquence (I), un tensioactif pulmonaire renfermant ce dernier et un mélange lipidique, ainsi qu'un remède contre le syndrome de souffrance respiratoire, renfermant ce tensioactif comme principe actif. Dans ladite séquence, Xaa désigne l'absence de n'importe quel élément ou bien représente Cys, Ser ou Ala; Xbb représente Cys, Ser ou bien Ala; Xcc représente His ou Asn; Xdd représente Leu ou Ile; Xee représente Val ou Ile; Xff représente Ile, Leu ou bien Val; et Xgg désigne l'absence de n'importe quel élément ou bien représente Leu. Ce peptide est facile à isoler et à purifier, peut être produit en série dans un court délai et possède une puissante activité de surface lorsqu'il est mélangé avec un mélange lipidique. Le tensioactif pulmonaire présente une bonne activité d'interruption ainsi qu'une puissante activité de surface, et il est ainsi utile comme remède contre le syndrome de souffrance respiratoire.
PCT/JP1993/000492 1992-04-17 1993-04-16 Peptide synthetique, tensioactif pulmonaire le renfermant, et remede contre le syndrome de souffrance respiratoire Ceased WO1993021225A1 (fr)

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JP51818893A JP3376582B2 (ja) 1992-04-17 1993-04-16 合成ペプチド、それを含有する肺サーファクタント及び呼吸窮迫症候群治療剤

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JP4098083A JPH05294996A (ja) 1992-04-17 1992-04-17 合成ペプチド、それを含有する肺サーファクタント及び呼吸窮迫症候群治療剤
JP4/98083 1992-04-17

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WO1993021225A1 true WO1993021225A1 (fr) 1993-10-28

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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1995015980A1 (fr) * 1993-12-08 1995-06-15 Tokyo Tanabe Company Limited Nouveau peptide synthetique, tensioactif pulmonaire contenant ledit peptide et remede contre le syndrome de souffrance respiratoire
WO1995032992A1 (fr) * 1994-05-31 1995-12-07 Byk Gulden Lomberg Chemische Fabrik Gmbh Analogues peptidiques de synthese de la proteine sp-c de surfactant
RU2144925C1 (ru) * 1993-12-08 2000-01-27 ТТ Фармасьютикалс, Инк. Новые синтетические пептиды, легочная поверхностно-активная композиция, лекарственный препарат для лечения респираторного дистресс-синдрома
JP2002529394A (ja) * 1998-11-10 2002-09-10 ビイク グルデン ロンベルク ヒエーミツシエ フアブリーク ゲゼルシヤフト ミツト ベシユレンクテル ハフツング 肺サーファクタント組成物を含有する治療セット
US6737243B1 (en) 1998-07-24 2004-05-18 Altana Pharma Ag Determination of the hydrophobic pulmonary surfactant protein SP-C

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* Cited by examiner, † Cited by third party
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DK0652227T3 (da) * 1993-04-30 1998-12-21 Tt Pharmaceuticals Inc Fremgangsmåde til oprensning af et hydrofobt polypeptid
WO1996017872A1 (fr) * 1994-12-07 1996-06-13 Tokyo Tanabe Company Limited Intermediaire pour la production d'un peptide surfactant

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JPS63503222A (ja) * 1986-05-06 1988-11-24 サイオス　ノバ　インコーポレイテッド 肺の疎水性界面活性物質に結合した分子量６，０００ダルトンのタンパク質及びその多量体
JPH01501282A (ja) * 1986-12-08 1989-05-11 ホイツトセツト，ジエフリー　エイ． 肺胞疎水性界面活性物質関連タンパク質

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JPS63503222A (ja) * 1986-05-06 1988-11-24 サイオス　ノバ　インコーポレイテッド 肺の疎水性界面活性物質に結合した分子量６，０００ダルトンのタンパク質及びその多量体
JPH01501282A (ja) * 1986-12-08 1989-05-11 ホイツトセツト，ジエフリー　エイ． 肺胞疎水性界面活性物質関連タンパク質

Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1995015980A1 (fr) * 1993-12-08 1995-06-15 Tokyo Tanabe Company Limited Nouveau peptide synthetique, tensioactif pulmonaire contenant ledit peptide et remede contre le syndrome de souffrance respiratoire
AU682738B2 (en) * 1993-12-08 1997-10-16 Mitsubishi-Tokyo Pharmaceuticals, Inc. Novel synthetic peptide, lung surfactant containing the same, and remedy for respiratory distress syndrome
US5827825A (en) * 1993-12-08 1998-10-27 Tokyo Tanabe Company Ltd. Synthetic peptide, lung surfactant containing the same and remedy for respiratory distress syndrome
RU2144925C1 (ru) * 1993-12-08 2000-01-27 ТТ Фармасьютикалс, Инк. Новые синтетические пептиды, легочная поверхностно-активная композиция, лекарственный препарат для лечения респираторного дистресс-синдрома
CN1057099C (zh) * 1993-12-08 2000-10-04 三菱东京制药株式会社 新的合成肽，其中间体及其生产方法
WO1995032992A1 (fr) * 1994-05-31 1995-12-07 Byk Gulden Lomberg Chemische Fabrik Gmbh Analogues peptidiques de synthese de la proteine sp-c de surfactant
AU690280B2 (en) * 1994-05-31 1998-04-23 Takeda Gmbh Synthetic peptide analogs of lung surfactant protein SP-C
US6737243B1 (en) 1998-07-24 2004-05-18 Altana Pharma Ag Determination of the hydrophobic pulmonary surfactant protein SP-C
JP2002529394A (ja) * 1998-11-10 2002-09-10 ビイク グルデン ロンベルク ヒエーミツシエ フアブリーク ゲゼルシヤフト ミツト ベシユレンクテル ハフツング 肺サーファクタント組成物を含有する治療セット

Also Published As

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JP3376582B2 (ja) 2003-02-10
JPH05294996A (ja) 1993-11-09
AU3905693A (en) 1993-11-18

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