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WO1993018065A1 - Substance proteique physiologiquement active - Google Patents

Substance proteique physiologiquement active Download PDF

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Publication number
WO1993018065A1
WO1993018065A1 PCT/JP1993/000294 JP9300294W WO9318065A1 WO 1993018065 A1 WO1993018065 A1 WO 1993018065A1 JP 9300294 W JP9300294 W JP 9300294W WO 9318065 A1 WO9318065 A1 WO 9318065A1
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WO
WIPO (PCT)
Prior art keywords
physiologically active
gly
active substance
chromatography
proteinaceous
Prior art date
Application number
PCT/JP1993/000294
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English (en)
Japanese (ja)
Inventor
Hajime Nawata
Fumio Umeda
Teruaki Yamauchi
Mitsunori Masakado
Original Assignee
Mochida Pharmaceutical Co., Ltd.
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 Mochida Pharmaceutical Co., Ltd. filed Critical Mochida Pharmaceutical Co., Ltd.
Publication of WO1993018065A1 publication Critical patent/WO1993018065A1/fr

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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
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
    • C07K14/4702—Regulators; Modulating activity
    • C07K14/4705—Regulators; Modulating activity stimulating, promoting or activating activity
    • 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 broth tag run Gin 1 2 (also known; prosulfuron evening cyclin (Pros ta cy c 1 in) , hereinafter referred to as PG 1 2) Novel proteinaceous raw physiologically active substance having a production stimulating activity, their preparation and
  • the present invention relates to a pharmaceutical composition containing the proteinaceous physiologically active substance.
  • novel proteinaceous physiologically active substance having an activity to promote the production of PG 1 2 from vascular endothelial cells.
  • Prostaglandin pros tagl and ins PG was published in 1930 by Kurzrock et al. [Pro S0c Exp. Biol. Med. 268, 1930], and was reported as a uterine muscle contractile active substance present in semen. Since then, basic research on prostaglandins has developed rapidly. Prostaglandin is a general term for biologically active substances based on prostanoic acid contained in human and animal tissues and organs.A series of chemicals called arachidonic acid cascades starting from arachidonic acid in cells. The reaction produces several prostaglandins with different chemical structures. It is.
  • prostaglandin plays an important role as a vasoactive substance for vascular smooth muscle [Pathophysiology, Vol. 2, p. 792, 1983], and thromboxane, an active prostanoid derived from platelets and blood vessel walls.
  • TXA 2 TXA 2
  • PGI 2 potent platelet aggregation inhibitory action and vascular relaxing action, and antagonism to create the TXA 2 platelet derived are involved in the maintenance of Homeosutashisu in vivo [Pre Tissue Jar Fungus, Narob Pharmacol. (Br. J. Parmac.), Vol.
  • the decrease in production may be attributable to the decrease in PG12 production stimulating activity present in the blood above [Metabolism] (Me t ab o 1 ism) 38, 837, 1989, Ha emo stasis 16, 447, 1986 and Diabetis Research and Clinical Practice (Diab. Res. Cl in. Prac. 3), p. 243, 1987].
  • PGI 2 As described above having a platelet aggregation inhibitory action, smooth muscle relaxing action, such as a blood vessel contact and bronchi besides that, have a gastric acid secretion inhibiting action and the like. Based on these physiological effects, it is conceivable to develop PGI 2 as a pharmaceutical product, but PGI 2 is a very chemically unstable substance with a neutral water of 37 and a half-life of 5 minutes. Absent. On the other hand, while maintaining the natural PG I 2-like activity, chemically stable PG 1 2 class ⁇ the anticoagulants, attempts to cane developed as pharmaceuticals such as a vasodilator has been made. However, it may be more desirable for the living body to make PGI 2 unstable and short.
  • the living body must always be ready to respond to PGI 2 that is produced promptly in an emergency, and by providing a large amount of stable PGI 2 analogs, the PGI 2 It is possible that responsiveness to PGI 2 may be reduced, making it impossible to respond to PGI 2 in an emergency. Indeed, pretreatment of prostaglandin E! (PGEa) and stable PGI 2 analogs due Examples of cells naturally occur to c AMP rise response to PG 1 2 no longer occurs has been reported [Prostaglandins (Pros tag 1 an nd ins.) 19, 2 pages, 1980].
  • PSA is present in blood, but its active substance (PSF) is It has not been identified and identification is desired.
  • PSF stimulates PG 12 production from endothelial cells, inhibiting platelet aggregation having the PG 1 2 by increasing the PG 12 concentration in the blood, smooth muscle relaxing action, to express gastric acid secretion inhibiting action, etc. it can.
  • PSF hemolytic uremic syndrome
  • thrombotic thrombocytopenic purpura peripheral artery occlusion
  • cardiac ischemia cerebral ischemia
  • atherosclerosis cerebral obstruction
  • hyperlipidemia diabetes, heart failure, angina It has the potential to be used in the treatment of sickness, ischemic heart disease, depressive heart disease, choroidal circulation disorder, bronchial disease, gastric ulcer, pregnancy and eclampsia. Disclosure of the invention
  • An object of the present invention is to provide a novel protein bioactive substance by purifying and isolating a substance that stimulates the production of PGI 2 from vascular endothelial cells in view of the above situation. Another object of the present invention is to provide a method for producing the same. It is a further object of the present invention to provide a pharmaceutical composition for the above-mentioned diseases based on the action of the proteinaceous physiologically active substance to stimulate PG12 production.
  • the present inventors have conducted intensive studies on PG12 stimulating activity present in the blood and in the culture supernatant of human cell lines in order to achieve the above-mentioned object, and found that the culture supernatant of normal human diploid fibroblasts It was confirmed that a substance having a PG12 production stimulating activity was present at a high concentration. Next, we succeeded in isolating and purifying a novel proteinaceous physiologically active substance having PGI 2 ⁇ stimulating activity from the culture supernatant, and determined its amino acid sequence.
  • the present invention provides a novel proteinaceous physiologically active substance having the following features.
  • the molecular weight is 30 to 36 kDa in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under non-reducing conditions.
  • the present invention uses at least one of the following steps (a) to (c) from a raw material containing the novel proteinaceous physiologically active substance, preferably a culture supernatant of normal human diploid fibroblasts.
  • a raw material containing the novel proteinaceous physiologically active substance preferably a culture supernatant of normal human diploid fibroblasts.
  • the present invention also provides a novel pharmaceutical composition for treating the following diseases, which comprises the novel proteinaceous physiologically active substance as an active ingredient.
  • FIG. 1 is a graph showing an elution pattern obtained by DEAE-5 PW anion exchange chromatography.
  • FIG. 2 is a graph showing an elution pattern obtained by heparin-5 PW affinity chromatography.
  • FIG. 3 is a graph showing the development pattern of protein-, gel-filtration and ram chromatography.
  • FIG. 4 is a schematic diagram showing an experimental result by SDS-PAGE. BEST MODE FOR CARRYING OUT THE INVENTION
  • the proteinaceous physiologically active substance of the present invention is a novel substance having at least one amino acid sequence represented by any of the following formulas (1) to (II).
  • amino acid sequence portion of any one of the above formulas (I) to (III) may be any portion of the proteinaceous physiologically active substance of the present invention.
  • the amino acid sequence portions of the above formulas (I) to (III) may be adjacent to each other or may contain an arbitrary amino acid sequence between them.
  • novel proteinaceous physiologically active substance has a molecular weight of 30 to 36 kDa in non-reducing conditions in sodium dodecyl sulfate sodium acrylamide gel electrophoresis (SDS-PAGE).
  • novel proteinaceous bioactive substances of the present invention have activity which act on vascular endothelial cells, stimulate the production of prostaglandin 1 2 (PG 1 2) from the cells. The method for measuring this activity will be described later.
  • the novel proteinaceous physiologically active substance of the present invention may be of any origin, but is preferably of human origin, more preferably of normal human diploid fibroblasts. Those produced in the culture supernatant of diploid fibroblasts are preferred. For example, it can be obtained by culturing normal human diploid fibroblasts or the like in an appropriate growth medium, collecting a culture solution having a PG production stimulating activity, and purifying a culture supernatant obtained. For culturing normal human diploid cells, a commonly used medium can be used, but Dulbecco's modified Eagle medium, RPMI164 medium, Eagle's minimum essential medium or ham medium is particularly preferable.
  • serum-free medium because the substance of the present invention is purified and isolated from the culture supernatant. It is desirable that the culture supernatant after completion of the culture is concentrated by, for example, a filter method or an ultrafiltration method.
  • the raw material containing the novel proteinaceous physiologically active substance of the present invention obtained as described above is purified by using at least one of the following steps.
  • the following steps (a), (b) and (c) ) May be performed according to a known protocol.
  • the order may be arbitrary, and may be repeated as necessary.
  • operations such as concentration, dialysis, etc. may be added between or before and after one step and the other, depending on the further, for example, chromatofocusing, adsorption chromatography, water chromatography, reverse phase Combination with other chromatography methods such as chromatography, thin-layer chromatography, partition chromatography, and other protein purification methods such as isoelectric focusing, polyacrylamide electrophoresis, and salting Is also good.
  • the collected fraction may be further subjected to reversed-phase high-performance liquid chromatography for the purpose of isolation, purification, purity confirmation, and the like.
  • appropriate elution conditions or developing conditions, eluate or developing solution may be selected in consideration of the properties of each chromatography carrier, target substance, contained contaminants, and the like.
  • the elution may be linear gradient elution or stepwise gradient elution.
  • the novel proteinaceous physiologically active substance of the present invention can be isolated from the culture supernatant preferably by the following method.
  • Culture supernatant of normal human diploid fibroblasts is subjected to anion exchange chromatography, for example, DEAE-5PW (manufactured by Tosoh I), onoQ (manufactured by Pharmacia) or Q-Sepharose column (manufactured by Pharmacia) And elute by sodium chloride concentration gradient method.
  • the PG 1 2 Stimulating activity of the eluted each fractions were measured to obtain fractions which are active. The measurement of the PGI 2 production stimulating activity will be described later.
  • this fraction is adsorbed to affinity chromatography, for example, heparin-15PW (manufactured by Tosoh Corporation) or the like, and eluted by a sodium chloride concentration gradient method.
  • the PGI 2 production stimulating activity is measured in the same manner, and the active fraction is collected. Next, this fraction is subjected to gel filtration chromatography, for example, Protein One Pack (Millipore Limited Waters Chromatography Division), Sephacryl S-100HR (Pharmacia) or TSK Gel G3000 SW XL column (Tosoichisha). Ltd.) and the like developed using, by Rukoto measuring the PG 1 2 production stimulating activity can be obtained biologically active fraction. This fraction is then subjected to affinity chromatography, for example, insulin-like growth factor (Insulin-like growth factor).
  • affinity chromatography for example, insulin-like growth factor (Insulin-like growth factor).
  • IGF e-like growth factor
  • the PGI 2 production stimulating activity can be measured by the following method.
  • vascular endothelial cells collected from the thoracic aorta membrane by a detachment method were subcultured in Dulbecco's modified medium containing 10% fetal serum, and cultured until they reached a saturated cell density. After adding the measurement sample and incubating for 60 minutes, the stable metabolite of PGI 2 in the supernatant is
  • O - Ke Bok - r F la (hereinafter, 6 - keto - PGF described as 1 alpha) can be obtained PG 1 2 production amount by the ⁇ a constant!.
  • a commercially available kit for measuring 6-keto PGF1 ⁇ may be used.
  • the amino acid sequence of the novel proteinaceous bioactive substances of the present invention with purified PG 1 2 production stimulating activity it may be used the following method.
  • the substance of the present invention can be directly analyzed by the sequencer, or peptide fragments obtained by enzymatic cleavage of trypsin, lysyl endopeptidase, etc. can be analyzed.
  • the sequence of each amino acid can be determined.
  • the novel proteinaceous physiologically active substance of the present invention is characterized by inhibiting platelet aggregation of PG I 2
  • the present invention provides a pharmaceutical composition containing the proteinaceous physiologically active substance of the present invention as at least one active ingredient for the following various diseases because it has an action, a smooth muscle relaxing action, a gastric acid secretion inhibitory action and the like.
  • Hemolytic uremic syndrome Hemolytic uremic syndrome, thrombotic thrombocytopenic purpura, peripheral arterial occlusion, cardiac ischemia, cerebral ischemia, arteriosclerosis, cerebral obstruction, hyperlipidemia, diabetes, heart failure, angina, ischemic heart disease, depression Hematologic heart disease, choroidal circulation disorder, bronchial disease, gastric ulcer, pregnancy eclampsia.
  • the dosage of the novel proteinaceous physiologically active substance of the present invention may vary depending on the patient's sex, age, body weight, disease type and its pathological condition, depending on the dosage form, but the effective dosage is 1 ng. 22 mg / kg, preferably 10 ng to 200; / g / kg.
  • the pharmaceutical form of the pharmaceutical composition of the present invention may be any as long as it can supply an effective amount to the disease lesion, and examples thereof include tablets, powders, powders, capsules, ointments, powders, and injections. Etc. are exemplified.
  • the pharmaceutical composition of the present invention contains a commonly used pharmaceutical mixture such as an excipient, a stabilizer ⁇ ) or a solubilizing agent, as long as the pharmacological properties are not impaired. May be.
  • Specific examples include Ringer's solution, phosphate buffer, human serum albumin, hydrolyzed gelatin, sucrose, dextran, polyethylene glycol, etc., which are appropriately selected and used depending on the drug form.
  • a culture supernatant was prepared and purified by the following method to obtain a novel proteinaceous physiologically active substance of the present invention.
  • Normal human diploid fibroblasts were prepared in 4.8 ⁇ 10 4 cells ZmL in Dulbecco's modified Eagle's medium containing 15% fetal bovine serum. 3 L of this cell suspension was inoculated in a rotary culture vessel, and 5% CO 2 -95% air 37. C. Three days after the cell implantation, the culture solution was replaced with 3 L of fresh Dulbecco's modified Eagle's medium containing 15% fetal calf serum, and the culture was continued for 2 days.
  • the culture solution was removed, and the cells were washed with a Ca2 + -and Mg2 + -free Dulbecco's monophosphate physiological saline solution, and 3 L of Dulbecco's modified Eagle's medium without phenol red was added. And cultured for 2 days. After recovering the culture, the cells were filtered through a 2.5 / zm filter (CN cartridge, 30 inch, manufactured by Millipore) to remove cell debris.
  • CN cartridge 2.5 / zm filter
  • a two-stage concentration using a hollow fiber module (Molsep fiber FS-106 kDa cut-off, manufactured by Daicel Chemical Industries, Ltd.) and a filtration filter (Omega mini-set, manufactured by Fuji Filter Industries, Ltd.) with a molecular weight cutoff of 10 kDa. It concentrated by operation.
  • This concentrated solution is used in a dialysis tube (stained with a molecular weight cutoff of 3.5 kDa
  • the solution was dialyzed against 2 OmM Tris-HCl buffer (pH 7.8) for 4 to 1 minute using the following method. After further dialysis with the same buffer at 4 e C for 8 hours, the mixture was sequentially filtered through 1.2 ⁇ m filter Yuichi (DF Assembly, Nippon Pall) and 0.22 zm filter-one system (Koining). did.
  • Culture supernatant of normal human diploid fibroblasts prepared according to (1) was pre-equilibrated with 2 OmM Tris-monohydrochloride buffer (PH7.8). DEAE-5 PW (Tosoichi) Anion exchange chromatography The mixture was adsorbed on a column, eluted with a linear gradient method of 0 to 1.0 M sodium chloride prepared using a 2 OmM Tris-monohydrochloride buffer (pH 7.8), and the absorbance was simultaneously measured at 280 nm.
  • the active fraction obtained in step 1 was dialyzed against 10 mM phosphate buffer (pH 7.4) and equilibrated with 1 OmM phosphate buffer (pH 7.4).
  • Heparin (HEPAR IN) -5PW E. Adsorbed on an affinity column and eluted with a linear gradient of 0 to 0 M sodium chloride prepared using 1 OmM phosphate buffer (pH 7.4). The absorbance was simultaneously measured at nm.
  • the eluted PGI 2 production stimulation activity of each off Rakushiyon measures, the fractions with the eluted PG I 2 production stimulating activity at sodium ⁇ beam chloride concentration of 450 to 500 mM were pooled (FIG. 2) o
  • the active fraction obtained in step 2 was concentrated with Centricon-10 (manufactured by Amicon), and then pre-equilibrated with 10 mM phosphate buffer (pH 7.4). It was developed using two connected gel filtration columns from Millipore Limited Waters Chromatography Division, and absorbance was simultaneously measured at 280 nm. Was measured PG 1 2 Stimulating activity of each fraction, activity was observed around a molecular weight 3 OKD a (FIG.
  • Fig. 3 o
  • the numbers in Fig. 3 indicate molecular weight (kDa), Vt indicates the total volume of the filler, and Vo indicates the excluded volume.
  • the active fraction obtained in step 3 was applied to an IGF-affinity column equilibrated with 1 OmM phosphate buffer (pH 7.4).
  • IGF-affinity column a pressure-resistant ram in which recombinant IGF-I was bound as a ligand to AFFEPREP 10 (manufactured by Nippon Bio-Rad Laboratories) was used for this experiment. After adsorption, the sample was eluted with 0.5 M acetic acid. Where was measured PG 1 2 Stimulating activity, PG 1 2 surname stimulating activity was found in non-adsorbed surface min.
  • the molecular weight of the novel proteinaceous physiologically active substance of the present invention obtained in step 5 of Example 1 was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under non-reducing conditions. A single band was observed at a molecular weight of 33 kDa (Fig. 4).
  • Figure 4 is a schematic diagram of a reference photograph after SDS-PAGE electrophoresis.
  • lines A and C are bands of a standard sample having a known molecular weight
  • line B is a band of the novel proteinaceous bioactive substance of the present invention.
  • the numbers represent the molecular weight (kDa) of the standard sample.
  • the novel proteinaceous physiologically active substance of the present invention obtained in step 5 of Example 1 was digested with trypsin to form a peptide fragment.
  • Formula (I) Formula (I)
  • Vascular endothelial cells were collected from the thoracic aorta intima by an isolation method. The obtained vascular endothelial cells were then placed in a Dulbecco's modified Eagle's medium containing 10% OU / mL penicillin and 100 gZmL streptomycin containing 10% fetal calf serum, at 37 ° C under 5% CO 2 -95%. The cells were subcultured in C. The medium was changed twice a week, and after passage 5-10, vascular endothelial cells were treated with 0.05% trypsin.
  • vascular endothelial cells were transferred to a 24-well culture dish containing 1 mL of Dulbecco's modified Eagle's medium containing 10% fetal bovine serum, and cultured to 5 ⁇ 10 4 cells Z well.
  • Dulbecco's modified Eagle medium 500 containing the following various samples, the mixture was incubated at 37 ° C for 60 minutes.
  • the PG12 production stimulating activity of the proteinaceous physiologically active substance of the present invention increased in a concentration-dependent manner.
  • Example 4 A toxicity test in mice was carried out using the proteinaceous physiologically active substance of the present invention obtained in Example 1. That is, the proteinaceous physiologically active substance obtained in Example 1 was intravenously administered once daily for one week to five male and female 6-week-old ICR mice once a day. Administration period The general characteristics of each animal were observed throughout. The highest dose group in this study
  • Example 6 10 Omg of the proteinaceous physiologically active substance of the present invention obtained in Example 1 was dissolved in 10 OmL of a physiological saline solution containing 1 Omg / mL of hydrolyzed gelatin, and a filter having a pore size of 0.22 m (Mirex GV Filtration and sterilization were carried out using Millipore Corporation. This was aseptically dispensed in 2 mL aliquots into glass vials, sealed and sealed for injection lysate. (Example 6)
  • Example 2 10 Omg of the proteinaceous physiologically active substance of the present invention obtained in Example 1 was dissolved in 10 OmL of 1 OmM PBS (pH 7.4) containing 10 Oiiig / mL of human serum albumin, and the pore size was 0.22 m. And sterilized by filtration using a filter (Mirex GV Millipore). This was aseptically dispensed into glass vials in 3 mL portions, freeze-dried and sealed to give a freeze-dried preparation for injection. Availability of S ⁇
  • the novel proteinaceous bioactive substances of the present invention acts on vascular endothelial cells has the effect of stimulating the PG 1 2 of flavor from the cells. Therefore, based on the inhibitory action of PG12 on platelet aggregation, smooth muscle relaxation, gastric acid secretion, etc., hemolytic uremic syndrome, thrombotic thrombocytopenic purpura, peripheral arterial occlusion, cardiac ischemia, cerebral ischemia, arteriosclerosis, brain It can be expected to be used as a therapeutic drug for obstruction, hyperlipidemia, diabetes, heart failure, angina pectoris, ischemic heart disease, depressive heart disease, choroidal circulatory disorder, bronchial disease, gastric ulcer, pregnancy eclampsia, etc. .

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  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Zoology (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Toxicology (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Peptides Or Proteins (AREA)

Abstract

Nouvelle substance protéique physiologiquement active possédant au moins une séquence d'acides aminés représentée par n'importe laquelle des formules (I), (II) ou (III), et présentant un poids moléculaire compris entre 30 et 36 kDa, tel que déterminé par le procédé SDS-PAGE dans des conditions non réductrices. Ladite substance intervient sur les cellules hémangioendothéliales de manière à stimuler la production par celles-ci de prostaglandine I2(PGI2), et est dont utilisable comme composition pharmaceutique à usage thérapeutique.
PCT/JP1993/000294 1992-03-10 1993-03-10 Substance proteique physiologiquement active WO1993018065A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP5165092 1992-03-10
JP4/51650 1992-03-10

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WO1993018065A1 true WO1993018065A1 (fr) 1993-09-16

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5534615A (en) * 1994-04-25 1996-07-09 Genentech, Inc. Cardiac hypertrophy factor and uses therefor
US5571675A (en) * 1994-04-25 1996-11-05 Genentech, Inc. Detection and amplification of candiotrophin-1(cardiac hypertrophy factor)
US6472585B1 (en) 1994-04-25 2002-10-29 Genentech, Inc. Cardiotrophin-1 defective mouse
US7258983B2 (en) 1994-04-25 2007-08-21 Genentech, Inc. Cardiotrophin-1 compositions and methods for the treatment of tumor

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
CHEMICAL ABSTRACTS, Vol. 106, No. 23, 8 June 1987, (Columbus, Ohio, U.S.A.), T. INOGUCHI et al., "Abnormality of Serum Stimulatory Activity on Prostacyclin (PGI2) Production in Diabetics", page 533, Abstract No. 193982k; & DOMYAKU KOKA, 14(5), 1039-45, (Japan). *
CHEMICAL ABSTRACTS, Vol. 111, No. 11, 11 September 1989, (Columbus, Ohio, U.S.A.), S. MUROTA et al., "A Kallikrein-Induced New Peptide Stimulating Prostacyclin Production by Vascular Endothelial Cells", page 91078, Abstract No. 91080a; & ADV. PROSTAGLANDIN, THROMBOXYNL, LEUKOTRIENE RES., 19(Taipei Conf. Prostaglandin Leukotrine Res., 1988), 244-7, (Eng.). *
CHEMICAL ABSTRACTS, Vol. 111, No. 9, 28 August 1989, (Columbus, Ohio, U.S.A.), Y. UCHIDA et al., "Isolation and Identification of Human Plasma Factor which Stimulates Prostaglandin I2 Production and its Chainges in Patients with Acute Myocardial Infarction", page 542, Abstract No. 75679x; & PROSTAGLANDIN, THROMBOXANE, LEUKOTRIENE RES., 19 (Taipei Conf. Prostaglandin Leukotriene Res., 1988), 299-302, (Eng.). *
CHEMICAL ABSTRACTS, Vol. 116, No. 23, 8 June 1992, (Columbus, Ohio, U.S.A.), D.W. BRANCH et al., "Sera from Preeclamptic Patients Contain Factor (S) that Stimulate Prostacyclin Production by Human Endothelial Cells", page 601, Abstract No. 233209d; & PROSTAGLANDINS, LEUKOTRIENES ESSENT. FATTY ACIDS, 45(3), 191-5, (Eng.). *

Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5534615A (en) * 1994-04-25 1996-07-09 Genentech, Inc. Cardiac hypertrophy factor and uses therefor
US5571675A (en) * 1994-04-25 1996-11-05 Genentech, Inc. Detection and amplification of candiotrophin-1(cardiac hypertrophy factor)
US5624806A (en) * 1994-04-25 1997-04-29 Genentech, Inc. Antibodies to cardiac hypertrophy factor and uses thereof
US5627073A (en) * 1994-04-25 1997-05-06 Genentech, Inc. Hybridomas producing antibodies to cardiac hypertrophy factor
US5679545A (en) * 1994-04-25 1997-10-21 Genentech, Inc. Gene encoding cardiac hypertrophy factor
US5723585A (en) * 1994-04-25 1998-03-03 Genentech, Inc. Method of purifying cardiac hypertrophy factor
US6117650A (en) * 1994-04-25 2000-09-12 Genentech, Inc. Assay for cardiac hypertrophy
US6472585B1 (en) 1994-04-25 2002-10-29 Genentech, Inc. Cardiotrophin-1 defective mouse
US7258983B2 (en) 1994-04-25 2007-08-21 Genentech, Inc. Cardiotrophin-1 compositions and methods for the treatment of tumor

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