[go: up one dir, main page]

WO1985005618A1 - Procede de preparation de derives de l'interferon - Google Patents

Procede de preparation de derives de l'interferon Download PDF

Info

Publication number
WO1985005618A1
WO1985005618A1 PCT/JP1984/000292 JP8400292W WO8505618A1 WO 1985005618 A1 WO1985005618 A1 WO 1985005618A1 JP 8400292 W JP8400292 W JP 8400292W WO 8505618 A1 WO8505618 A1 WO 8505618A1
Authority
WO
WIPO (PCT)
Prior art keywords
lys
ser
asn
arg
leu
Prior art date
Application number
PCT/JP1984/000292
Other languages
English (en)
Japanese (ja)
Inventor
Masakazu Kikuchi
Tsutomu Kurokawa
Susumu Honda
Original Assignee
Takeda Chemical Industries, 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 Takeda Chemical Industries, Ltd. filed Critical Takeda Chemical Industries, Ltd.
Priority to PCT/JP1984/000292 priority Critical patent/WO1985005618A1/fr
Priority to PCT/JP1984/000434 priority patent/WO1985005619A1/fr
Priority to ZA853966A priority patent/ZA853966B/xx
Priority to IL75302A priority patent/IL75302A0/xx
Priority to AU43219/85A priority patent/AU4321985A/en
Priority to DK248285A priority patent/DK248285A/da
Priority to PH32351A priority patent/PH22613A/en
Priority to JP60121787A priority patent/JPS615096A/ja
Priority to EP85106885A priority patent/EP0166993A3/fr
Priority to KR1019850003950A priority patent/KR860000377A/ko
Publication of WO1985005618A1 publication Critical patent/WO1985005618A1/fr
Priority to US07/129,947 priority patent/US4855409A/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • 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
    • C07K14/52—Cytokines; Lymphokines; Interferons
    • C07K14/555—Interferons [IFN]
    • C07K14/57—IFN-gamma
    • 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 a novel polypeptide useful as a pharmaceutical product and a method for producing the same.
  • IFN-r r-type interferon
  • N-r is said to have higher anti-cell proliferative activity and anti-tumor activity than IFN-naya-, and is expected from clinical applications.
  • IFN-naya- anti-cell proliferative activity and anti-tumor activity than IFN-naya-
  • the present inventors consider that the above-mentioned disadvantages of the known rIFN-r are based on the two Cys present at the N-terminal thereof, and are equivalent to or higher than that of rlFN-r.
  • the present invention has been completed by making an active effort in the production of a novel oligopeptide having biological activity and hardly causing dimerization or multiplication.
  • the present invention uses the formula
  • X is preferably a bond.
  • Y is preferably Cys-Gin, Gin or Gin, and particularly preferably Gin.
  • Z is Lys or Lys—Arg—Lys—Arg—Ser—Gln-Met-Leu-Pe—Arg-Glj—Arg (E or Lys—Arg—Lys—Ar—Ser—G1n—Met — Le u— P he— A and g—G 1 y—A r—A rg—A 1 a-Ser-Gln (I).
  • X is a bond
  • Y is Cys-Gln- or Gin
  • Z is Lys or peptide (IT).
  • the polypeptide (I) has, for example, ATG at the 5 ′ end and has a downstream
  • Trp Lys Glu Glu Ser Asp Arg Lys lie Met Gin Ser Gin lie Val Ser Phe Tyr Phe Lys Leu Phe Lys
  • the region encoding the polypeptide (1) may be any as long as it has the nucleotide sequence encoding the above-mentioned polypeptide ( ⁇ ).
  • AGT CAG ATG CTG TTT CGA GGT exchange CGA AGA GCA TCC CAG (3 in DN Alpha can be mentioned that represented by the nucleotide sequence counted from the 5 'end indicates a 3 ⁇ 4 Ru nucleotide sequence from 1 to 1 6 codons] where indicated, the divided TGC as Y 1 CAG or CAG mosquito, preferably having a base sequence represented by AAG CGA AAA AGG AGT CAG ATG CTG TT CGA GGT CGA AGA GCA TCC CAG (V) as Z 1.
  • Examples of translation termination codons include TAA, TGA and TAG. And 3) TAA is preferred.
  • the DNA is a signal chain between the ATG at the 5 'end and the DNA encoding the polypeptide (: T).
  • the above-mentioned DNA preferably has a promoter upstream of ATG, and the promoter may be any promoter as long as it is an appropriate promoter corresponding to a host used for production of a transformant. .
  • E. coli Esscherichia coli; Example, 29 4, W 3110, DH 1 3 ⁇ 4 etc.
  • trp promoter one, lac promoter, rec A promoter - coater, Li 3 Mr. promoter foremost, etc.
  • Bacillus subtilis Bacillus subtilis (Baci- llus subtilis (eg, M ⁇ 114), SP01 promoter, SP02 promoter, penP promoter, etc.
  • yeast Sacha-romyces cerevisiae; AH22, etc.
  • PH05 promoter — eg.
  • PG promoter GAP.
  • animal cells eg, monkey cell C0S-7, Chinese hamster cell CH0, etc.
  • promoters derived from SV40, etc. are mentioned. It is preferable to use the trp promoter using E. coli as a host.
  • DNA having ATG at the 5 'end and a polypeptide (downstream coding region), followed by a translation stop codon downstream of the ATG is obtained from known IFN-r produced by chemical synthesis or genetic engineering. by processing cDNA or chromosome-derived IFN-r DNA] 9.
  • the transformant of the present invention can be prepared by subjecting the working plasmid obtained as described above to a method known per se [Cohen SN et al., Pro. Natl. Acad. Sci. 6 ⁇ , 2110 (1972)].
  • the polypeptide (I) can be produced by culturing the above transformant, producing and accumulating the polypeptide (I) in the culture, and collecting this.
  • Examples of the medium include M9 medium containing glucose and casamino acid [Miller, J. Experiments in Molecular Genetics, 431-433 (Cold Spring Horbor Laboratory, New York, 1972)].
  • a drug such as 3
  • Culture is usually performed at 15 to 43 ° C. for 3 to 24 hours, and if necessary, aeration and agitation can be added.
  • the culture is performed at a low temperature of about 30 ° C to 36 ° C, and the inactivation of the ⁇ repressor is performed at about 3 ° C. It is preferable to carry out at 7 ° C to 42 ° C. Also, in order to make the recA promoter work more efficiently, that is, to reduce the function of suppressing the recA gene expression, mitomycin C and any drug such as naldixic acid were added as necessary. be able to.
  • the cells After culturing, the cells are collected by a known method, suspended in a buffer, for example, and then lysed with a protein denaturant, sonicated, or treated with an enzyme such as lysozyme. To obtain a supernatant.
  • the polypeptide (I) in which Z is a polypeptide or an amino acid residue having an amino acid of 15 or less has a large number of amino acids in the same Z] 9.
  • Z is a peptide having all 16 amino acids
  • It can also be produced by culturing a transformant containing NA and purifying the transformant under conditions that facilitate the use of protease.
  • the polypeptide (I) can be isolated from the obtained supernatant by a generally known protein purification method.
  • the antibody having binding ability to IFN-r or polypeptide (I) can be advantageously purified using its antibody ram.
  • H-Lys-Arg-Lys-Arg-Ser Gin—Met—Leu—Phe—Arg—Gly—rg—Arg—Ala—Ser—Gin—OH
  • Example 12 (r2—11.1 monoclonal antibody ram), Example 18 (r3—11.1 monoclonal) of JP-A-59-80664 Antibody column prepared from antibody) as described above) or ⁇ Gln—Asp—Pro—Tyr—Val—LyS—Glu—Ala—Glu—Asn—Leu—Lys—Lys—yr—Pe—Asn-Ala- Antibody potency of a monoclonal antibody against a peptide represented by Gly-OH [Japanese Patent Application No. 58-215168 (Patent on Jan. 15, 1983) WNr2-76.5 3 Monoclonal antibody column of antibody))]
  • libeptide (I) For purification with the above antibody column, for example, e.
  • the substance containing libeptide (I) is dissolved in a buffer solution near neutral, for example, a phosphate buffer or a tris-hydrochloride buffer, and adsorbed to the antibody buffer.
  • a buffer solution near neutral for example, a phosphate buffer or a tris-hydrochloride buffer
  • the polypeptide (I) is eluted.
  • Eluents include weakly acidic solutions such as acetic acid solutions, solutions containing polyethylene glycol, and e.
  • a solution containing a peptide that binds more easily than a peptide (I)], a high-concentration salt solution, a solution containing a combination of these, and the like are used. Those which do not accelerate the decomposition of the peptide (I) are preferred.
  • the column eluate is neutralized with a buffer according to a standard method. If necessary, i9 can be subjected to the above-described purification operation using the antibody force column again.
  • polypeptide (I) when Y is Cys-Gln, Gln or a bond, the polypeptide may be obtained as a mixture of a polypeptide in which X is a bond and a polypeptide in which X is Met. .
  • N-terminal amino acid of polypeptide (I) when the N-terminal amino acid of polypeptide (I) is Gin, it may be obtained as a mixture with the polypeptide (I) which is Gin.
  • the mixture can be used as it is for the purposes described below.
  • heating or treatment with a weak acid eg, dilute acid
  • Amino acids can lead to polypeptides (I) where ⁇ G ln.
  • the polypeptide (I) solution obtained here is subjected to dialysis, and if necessary, this can be made into a powder by lyophilization.
  • stabilizers such as sonorebitone, mannitonole, dextroses, manoleose, and quinose can be added.
  • the polypeptide (I) thus obtained has only one or no Cys, it is obtained as a stable monomer which is less susceptible to dimerization or multimerization than conventional rIFN-r, for example, Since precipitation hardly occurs in the concentration operation, and the biological activity thereof is hardly reduced with time, it can be advantageously used as a pharmaceutical or the like.
  • Polypeptides of the present invention (I) is purified to a polypeptide having a 1 0 7 u more specific activity in Virus activity measurement by cytopathic effect inhibition test of vesicular stomatitis virus against human amnion-derived WISH cells (VSV) And the known rIFN-r [Gray, PW et al., Supra] and
  • the voriveptide (I) of the present invention exhibits anti-Winores, anti-tumor, cell growth inhibitory and immunopotentiating effects.
  • the polypeptide (I) of the present invention can be mixed with sterile water, human serum anolebumin (HSA), physiological saline or other known physiologically acceptable carriers, and administered parenterally or topically.
  • HSA human serum anolebumin
  • 100,000 to 100 million units, preferably 400,000 to 400,000 units per day for an adult can be administered intravenously or intramuscularly.
  • Formulations containing the voriveptide (I) of the present invention may include other physiologically acceptable salts such as salts, diluents, adjuvants, other carriers, buffers, binders, surfactants, and preservatives. It may also contain active ingredients. Preparations for parenteral administration include sterile aqueous solutions or suspension samples in physiologically acceptable solvents, or sterile powders (usually polypeptides) that can be used in serial dilution with a physiologically acceptable diluent. (I) obtained as an sample (obtained by freeze-drying a solution).
  • physiologically acceptable salts such as salts, diluents, adjuvants, other carriers, buffers, binders, surfactants, and preservatives. It may also contain active ingredients. Preparations for parenteral administration include sterile aqueous solutions or suspension samples in physiologically acceptable solvents, or sterile powders (usually polypeptides) that can be used in serial dilution with
  • polypeptide (I) of the present invention IFN- a, ⁇ FN - 9 or IE 1 N-r or as re Nhoka Lee down such I Ntaroi Kin 2 of the present invention a 3 ⁇ 4 active ingredient It may be contained at 1-99% with respect to the polypeptide (I).
  • drawings and claims when amino acids, peptides, protecting groups, active groups, etc. are abbreviated, they are referred to as IUPAC-
  • the method of setting the cut as the anti-inoles activity (IFN-r activity eff) of the polypeptide was carried out as follows:
  • the international standard IFN- ⁇ defined by Unitit and crude derived from leukocytes IFH-r was measured using the VSV cytopathic effect inhibition test against human amniotic membrane-derived FL cell line, and from the comparison of the titers, the titer of leukocyte-derived crude N-r was determined to obtain a standard IFN- ⁇ standard.
  • the standard IFN- r was always arranged, and the assay was performed using the WISH-VSV system described above, and the titer was calculated from the ratio.
  • FIGS. 1, 2 and 3 show the plasmids shown in Examples 1 (i), (ii) and ( ⁇ ), respectively, pH IT trpl l O l— d2, pHIT trp 1201-d 3 The construction of pH IT trp 120 l-ci4 is shown.
  • IFN-expression plasmid PHIT trp 1101 [see Japanese Patent Application Laid-Open No. 59-80664, Reference Example 2 (ill)] was digested with restriction enzymes Avail and Pstl, and IFN-r gene was expressed. An Avail-; Pstlkb DNA fragment containing the portion was isolated. This DNA fragment contains a protein synthesis initiation codon chemically synthesized by the above-mentioned ester method.
  • Plasmid ptrp 771 [see Reference Example 2 (II) above] was digested with the restriction enzymes Clal and Pstl to obtain an IFN-r gene in which the above adapter was bound to the downstream of the trp promoter of a DNA fragment. And enter the DNA [IV; 1 is TGCCAG 1 has a base sequence (V)]], and polypeptide (:); ⁇ is Cys-Gin; Z is an expression plasmid encoding a peptide ():] pH IT trp 1101 —D 2 was constructed (Fig. 1).
  • IFN-r expression plasmid pHITtrp1101 was digested with restriction enzymes Avail, ⁇ > and Aval— containing the IFN-r gene portion; Pst 1 kb DITA fragment was obtained. To separate. An oligonucleotide adapter containing an initiation codon for protein synthesis synthesized by the above-described triestenolation method to this DNA fragment
  • GTACGTCCTG is ligated to the back of AvaE using T4 DNA ligase.
  • the current vector ptrp701 (see Reference Example 2 (() above) is digested with the restriction enzyme EcoRI, then partially degraded with Clal, and the resulting portion is treated with DNA e.
  • the DNA repaired with remerase I large fragment was circularized using T4 DNA ligase, the Cla I recognition site closer to the EcoRI recognition site was broken, and the heterologous gene insertion site became the EcoRI site.
  • the IFN one r gene by binding the adapter be attached using the T 4 D if A ligase to ⁇ downstream of Toriputo Fuanpuromo one coater of DNA fragments obtained by cutting the ptr P 781 with the restriction enzymes EcoRI and Pstl Yo! ), DNA [IV; ⁇ said CAG, Z 1 is the nucleotide sequence (V)] have, polypeptide [gamma; however is Gin, Z Habe peptide (H)] encoding the expression plasmids FpH ITtrp l 201—d3 can be constructed ( Figure 2).
  • Escherichia coli 294 is transformed with this plasmid pHIT trp1201-d3 according to the method of Cohen et al. (Supra) to obtain a strain E. coli294 / pHIT trp120l- "d3 containing this plasmid.
  • Example l (i) an IF- r expression plasmid pH IT trpl l O l- was digested with restriction enzymes Avail and Pstl, and an Avail-Pstl 1 kb DNA fragment containing the IFN-r gene portion was digested. Fractionate. Digestion of this DNA fragment with the restriction enzyme AvaJT] produced a portion of the DNA fragment using DNA polymerase I large fragment, which was then chemically synthesized by the triester method. Oligonucleic acid containing start codon
  • Escherichia coli 294 was transformed with this plasmid pHI T trp 1201—d4 according to the method of Cohen et al. (Supra), and a strain containing this plasmid, E. coli 294 / p HIT trp 1201 to d4, was transformed into can get.
  • Plasmid containing strain constructed in Example 1 E. C011294 / pHI Ttrl 1101-d2 containing 89 / ml tetracycline, 0.4% casamino acid, 1% gnorecose The cells were cultured at 37 ° C using M9 medium, and when the growth reached KU220, 3 indolylacrylic acid / leic acid (IAA) was added to 25 / i to further culture for 4 hours. After culturing, the cells were collected by centrifugation, and this was replaced with 0.05M Tris containing 1Z10 of 10% sucrose ⁇ -. -Suspended in HC1 pH 7.6.
  • This lysate is centrifuged at 2000 rpm at 4 ° C in a 200 rpm rp ni (Serval centrifuge SS — 34 rotor) for 30 minutes, and the polypeptide [I; ⁇ , X is a bond or (and) Met , Y was Cys-Gin, and Z was Zuma. (IT)].
  • the antiviral activity of this supernatant was determined to be 2.87 ⁇ 10 8
  • Tris-HCl buffer (p H 7 ⁇ 0) 18 suspended in W, 4 obtained in the supernatant 20 »centrifuged 30 minutes at 10,000 X After 1 hour ⁇ ft 0 1 3 7 to the supernatant A buffer solution (pH 7.4) consisting of mM sodium chloride, 2.7 mM sodium chloride, 8.1 mM sodium phosphate and 1.5 mM phosphate monobasic (pH 7.4) (hereinafter abbreviated as PBS). The mixture was diluted with 260 2 and applied to an antibody ram (Mor2-11.1, column volume 12 ml) at a flow rate of 1 min.
  • PBS mM phosphate monobasic
  • the polypeptide obtained here [I; X is a bond or (and) Met, Y is Cys-Gin, Z is peptide ()] is 5.9 ⁇ ])
  • the specific activity is (1.0 10 7 U / a ⁇ ).
  • Laemmli Nature, 227, 680-685, (1970)]
  • mature rIFNr US Pat. (Filing date, September 20, 1983) See the specification]
  • a protein band was detected at a position showing almost the same mobility (molecular weight of about 18,000).
  • a slight protein band was observed at a position corresponding to the molecular weight of the dimer. 3 ⁇ 4 That is, the formation of Benimer was much less than that of the conventional rIFN-r.
  • Example 2 The frozen cells obtained by the methods of ( ⁇ ) and (iii) were each mixed with 0.1 M Tris-HCl containing 7 M guanidine hydrochloride and 2 mM phenylmethyi / resulfonyl fluoride. The suspension is suspended in 3 volumes of buffer (pH 7.0), stirred at 4 for 1 hour, and then centrifuged at 100,000 X for 30 minutes to obtain a clear supernatant. The supernatant is diluted 14-fold with P33 and applied to an antibody column (Mor 2-111).
  • buffer pH 7.0
  • the ram is then washed with 20 mM sodium phosphate buffer (pH 7.0) containing 0.5% guanidine hydrochloride, and then 20 mM solution containing 2M guanidine hydrochloride. Elution with sodium phosphate buffer (pH 7.0) yields a fraction having antiviral activity.
  • This fraction was previously purified with a 25 mM ammonium acetate buffer solution containing 1 mM ethylenediaminetetraacetic acid, 0.15 M sodium chloride, 10 mM cysteine and 2 M guanidine hydrochloride (The mixture is applied to a column of Sephacrylinole S-200 (manufactured by Bual Macia) equilibrated at pH 6.0) and eluted with the same buffer to obtain a fraction having anti-dise activity.
  • polypeptide [I; X is a bond or (and) Met, Y is Gin or (and) Gin, Z is peptide (II)] and polypeptide [I; X is a bond Or (and) the specific activity of Met, Y is a bond, and Z is a peptide (II)], which is equal to or higher than the polypeptide (I) obtained in Example 3 (i).
  • Example 2 Twenty-five frozen cells obtained by the methods (i), (ii) and (iii) were each added to a 1.5-fold amount of 0.15 M sodium borate buffer (pH 9.5). After suspending and stirring at 4 ° C for 1 hour, it is extracted by applying ultrasonic waves 5 times for 30 seconds each to 30 000 0 0? Centrifuge for 1 hour to obtain supernatant. This supernatant is mixed with Siri gel 25, previously washed with PBS, and mixed with 4! Stir gently.
  • the silica gel is packed into a column, washed with 20 to 30 times the volume of 1 M NaCl, and then with a 0.01 M sodium borate buffer (pH 0.5) containing 0.5 M tetramethyltinoleammonium chloride. Elution at 8.0) yields about 20 fractions showing antiviral activity. This was further divided into four fractions, each of which was applied to a monoclonal antibody (Mor 2-11.1) abundance column equilibrated with PBS, washed with 10 volumes of PBS, and then 50% ethylene glycol and Elute with 20 mM sodium phosphate buffer (pH 7.0) containing 1 M sodium chloride. Antiviral activity is eluted in the first about 20 minutes.
  • the above 15 Kd corresponds to the following: 'each is a polypeptide [I; X is a bond or (and) Met, Y is Cys-Gin, Z is Lys], and a polypeptide [I; Hand or (and) Met, Y is Gin or (and) Gin, Z is Lys] and polypeptide [I; X is a bond or (and) Met, Y is a bond, Z is Lys], Those corresponding to 17 Kd are polypeptides [I; where X is a bond or (and) Met, Y is Cys-Gin, Z is peptide (I)], polypeptide [I; X is a bond or (and) Met, Y is Gin or (and) Gin, Z is a peptide.
  • the peptide [I; X is a bond or (and) Met, Y is a bond, and Z is a peptide (H):]. ,. Industrial applicability
  • the polypeptide (I) of the present invention has an antiviral, antitumor, and immunopotentiating effect and is stable, and thus can be advantageously used as a pharmaceutical or the like.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • General Health & Medical Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biochemistry (AREA)
  • Wood Science & Technology (AREA)
  • Biotechnology (AREA)
  • Biophysics (AREA)
  • Microbiology (AREA)
  • Plant Pathology (AREA)
  • Physics & Mathematics (AREA)
  • Toxicology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Medicinal Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Peptides Or Proteins (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Saccharide Compounds (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

Nouveau polypeptide possédant une activité égale ou supérieure à celle de l'IFN-gamma et subissant difficilement une dimérisation ou une polymérisation, un transformant contenant de l'ADN codant le nouveau polypeptide et procédé de préparation du nouveau polypeptide à partir d'un produit de culture du transformant. L'ADN codant le nouveau polypeptide peut être préparé à partir d'un plasmide connu contenant le cADN du gène de l'IFN-gamma, le PHITtrp 1101. Cet ADN est lié à un vecteur et introduit dans un hôte pour produire un transformant. Une colonne d'anticorps est utilisée pour purifier le polypeptide du produit de culture. Le nouveau polypeptide peut être utilisé comme agent antiviral et antinéoplastique.
PCT/JP1984/000292 1984-06-06 1984-06-06 Procede de preparation de derives de l'interferon WO1985005618A1 (fr)

Priority Applications (11)

Application Number Priority Date Filing Date Title
PCT/JP1984/000292 WO1985005618A1 (fr) 1984-06-06 1984-06-06 Procede de preparation de derives de l'interferon
PCT/JP1984/000434 WO1985005619A1 (fr) 1984-06-06 1984-09-11 Nouveau polypeptide et son procede de preparation
ZA853966A ZA853966B (en) 1984-06-06 1985-05-24 Polypeptide and production thereof
IL75302A IL75302A0 (en) 1984-06-06 1985-05-24 Novel polypeptide and production thereof
AU43219/85A AU4321985A (en) 1984-06-06 1985-05-31 Gamma interferon analogues
DK248285A DK248285A (da) 1984-06-06 1985-06-03 Polypeptider samt deres fremstilling og anvendelse
PH32351A PH22613A (en) 1984-06-06 1985-06-03 Novel polypeptide and production thereof
JP60121787A JPS615096A (ja) 1984-06-06 1985-06-04 新規ポリペプチドおよびその製造法
EP85106885A EP0166993A3 (fr) 1984-06-06 1985-06-04 Polypeptide et sa production
KR1019850003950A KR860000377A (ko) 1984-06-06 1985-06-05 폴리펩티드의 제조방법
US07/129,947 US4855409A (en) 1984-06-06 1987-11-23 Novel polypeptides and method of producing same

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/JP1984/000292 WO1985005618A1 (fr) 1984-06-06 1984-06-06 Procede de preparation de derives de l'interferon

Publications (1)

Publication Number Publication Date
WO1985005618A1 true WO1985005618A1 (fr) 1985-12-19

Family

ID=13818356

Family Applications (2)

Application Number Title Priority Date Filing Date
PCT/JP1984/000292 WO1985005618A1 (fr) 1984-06-06 1984-06-06 Procede de preparation de derives de l'interferon
PCT/JP1984/000434 WO1985005619A1 (fr) 1984-06-06 1984-09-11 Nouveau polypeptide et son procede de preparation

Family Applications After (1)

Application Number Title Priority Date Filing Date
PCT/JP1984/000434 WO1985005619A1 (fr) 1984-06-06 1984-09-11 Nouveau polypeptide et son procede de preparation

Country Status (4)

Country Link
JP (1) JPS615096A (fr)
KR (1) KR860000377A (fr)
WO (2) WO1985005618A1 (fr)
ZA (1) ZA853966B (fr)

Families Citing this family (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6936694B1 (en) 1982-05-06 2005-08-30 Intermune, Inc. Manufacture and expression of large structural genes
CA1310924C (fr) * 1986-04-24 1992-12-01 Francis P. Mccormick Systeme d'administration d'un medicament a l'aide de particules infectieuses
GB8619725D0 (en) * 1986-08-13 1986-09-24 Hoffmann La Roche Homogenous interferon fragments
JP2653061B2 (ja) * 1986-12-27 1997-09-10 武田薬品工業株式会社 新規ポリペプチドおよびその製造法
ATE212672T1 (de) * 1989-10-24 2002-02-15 Chiron Corp System zur freisetzung von infektiösen proteinen

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS5890514A (ja) * 1981-10-19 1983-05-30 ジエネンテツク・インコ−ポレイテツド ヒト免疫インターフエロンをコートするｄｎａ配列
JPS5951792A (ja) * 1982-02-22 1984-03-26 バイオジェン　インコーポレイテッド Ｄｎａ配列、組換ｄｎａ分子およびヒト免疫インタフエロン様ポリペプチドの製造方法

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4582800A (en) * 1982-07-12 1986-04-15 Hoffmann-La Roche Inc. Novel vectors and method for controlling interferon expression

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS5890514A (ja) * 1981-10-19 1983-05-30 ジエネンテツク・インコ−ポレイテツド ヒト免疫インターフエロンをコートするｄｎａ配列
JPS5951792A (ja) * 1982-02-22 1984-03-26 バイオジェン　インコーポレイテッド Ｄｎａ配列、組換ｄｎａ分子およびヒト免疫インタフエロン様ポリペプチドの製造方法

Also Published As

Publication number Publication date
KR860000377A (ko) 1986-01-28
WO1985005619A1 (fr) 1985-12-19
JPS615096A (ja) 1986-01-10
ZA853966B (en) 1986-01-29

Similar Documents

Publication Publication Date Title
JP2515308B2 (ja) ヒト免疫インタ−フエロン
EP0043980B1 (fr) Interferon A mature de leucocytes humains, procédé pour sa production microbienne, produits intermédiaires correspondents et compositions le contenants
US6482613B1 (en) Microbial production of mature human leukocyte interferons
US5096705A (en) Human immune interferon
CA1341569C (fr) Production microbienne d'interferon associe a des fibroblastes humains
JPH07135992A (ja) α−インターフェロンの調製方法
US5582824A (en) Recombinant DES-CYS-TYR-CYS human immune interferon
JP2566909B2 (ja) 新規遺伝子配列、それによつてコ−ドされるｉ型インタ−フエロンペプチドおよびそのインタ−フエロンを産生する微生物
US4855409A (en) Novel polypeptides and method of producing same
US4980455A (en) Novel polypeptides and production thereof
KR100360594B1 (ko) 인간 인터페론 알파의 발현 분비벡터 및 이를 이용한인터페론 알파의 생산 방법
Nakamura et al. Molecular cloning of feline interferon cDNA by direct expression
IE872160L (en) Recombinant immune interferon fragments
WO1985005618A1 (fr) Procede de preparation de derives de l'interferon
US5157004A (en) Polypeptides and production thereof
EP0126230A1 (fr) ADN et l'emploi de celui-ci
JPS5841849A (ja) ヒト白血球インタフエロン
CN102796197A (zh) 一种hsa-gcsf突变体及其制备方法
JPS61501430A (ja) 修飾ガンマ・インタ−フェロン、そのｉｆｎをコ−ドするｄｎａ配列およびそのｉｆｎの生産方法
NZ214261A (en) Human immune interferon: production by recombinant dna technology
JPS60222500A (ja) 新規ハイブリツドインタ−フエロン
PH26492A (en) Recombinant human immune interferon
BG61060B2 (bg) Човешки имунен интерферон
CS273182B2 (en) Method of expressive vector production capable to give mature human leucocytic interferon in transformed strain of escherichia coli by means of expression

Legal Events

Date Code Title Description
AK Designated states

Designated state(s): MC