WO1995031542A1 - Adn mutant recepteur de l'insuline humaine - Google Patents
Adn mutant recepteur de l'insuline humaine Download PDFInfo
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- WO1995031542A1 WO1995031542A1 PCT/JP1995/000906 JP9500906W WO9531542A1 WO 1995031542 A1 WO1995031542 A1 WO 1995031542A1 JP 9500906 W JP9500906 W JP 9500906W WO 9531542 A1 WO9531542 A1 WO 9531542A1
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- insulin receptor
- dna
- human insulin
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- mutated
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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
- C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
- C07K14/72—Receptors; Cell surface antigens; Cell surface determinants for hormones
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- 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 abnormalities in the insulin receptor structural gene in non-insulin-dependent diabetes mellitus (hereinafter referred to as NID DM), and more particularly to a mutant human insulin receptor DNA containing a mutation at a specific site and a fragment thereof.
- NID DM non-insulin-dependent diabetes mellitus
- NIDDMM is one of the most frequent genetic diseases in humans, which is thought to be caused by several genetic abnormalities and environmental factors such as obesity, stress and aging. It is said that there are about 500,000 patients in Japan at present, and in recent years it has been designated as the four major diseases next to cancer, stroke, and myocardial infarction, and measures are urgently needed. Because NIDDM usually improves symptoms and test results with diet and exercise, it is advisable to diagnose it early, preferably before the onset of symptoms. Currently, in the early stages of diagnosis of NID DM, 75 g of glucose is orally administered on an empty stomach, blood is collected every 30 minutes, and blood glucose is measured for 2 hours to diagnose complicated diabetes such as OGTT. In many cases, they cannot be detected without a test, and early detection by simple and reliable diagnostic methods and appropriate prevention of onset are required.
- NID DM insulin receptor substrate 1
- IMS-1 insulin receptor substrate 1
- GIS-1 insulin receptor substrate 1
- GCS-2 insulin receptor substrate 1
- Genes include glucose transporter Eve 2, glucokinase, and mitochondrial genes. The latter two genes were searched for abnormalities in NIDDM, but both were reported to be only about 1%. Mechanism group, 1993 interim report).
- the present inventors prepared chromosomal DNA from the blood of a typical Japanese NI DDM patient and elucidated the nucleotide sequence of the insulin receptor DNA in order to elucidate the relationship between human insulin receptor gene abnormalities and NI DDM.
- the inventors have found that qualitative abnormalities of insulin receptor exist in NID DM patients at a significant frequency, and have completed the present invention.
- FIG. 1 is a photograph of electrophoresis showing a part of the nucleotide sequence of the insulin receptor / 3 subunit exon 13 of five NI DDM patients.
- FIG. 2 is a photograph of electrophoresis showing a part of the nucleotide sequence of insulin receptor / 3 subunit exon 22 of eight patients with NIDDM.
- Insurin wild type (IR WT) and mutant (IR A831) receptor Shows binding (A) and receptor ostial synkinase activity (B, C).
- A IR WT (clone No. 12: 0—O; N 0.21: ⁇ ) or I
- PI 3- kinase p 85 subunit of (P 85) in COS cells were co-expressed in each transient to the wild-type (IR WT) or artificial mutant IR (IR A831 or IR C1334). 10 -.
- FIG. 5 shows the relationship between the onset of NI DDM and the mutation in a family with IR ei 334 mutation.
- A Family analysis with substitution of TAC (Ty r 1334 ) ⁇ TGC (Cy s 133 ). Squares represent men and circles represent women. Those painted in black are NI DDM patients, the arrow represents the person, and the hatched one represents the deceased.
- B Allele-specific hybridization. PCR products from insulin receptor exon 22 obtained from genomic DNA of three patients or normal individuals were separated on an agarose gel and transferred to nitrocellulose filters. After hybridization with a 32 P-labeled oligonucleotide probe specific for the mutant allele, the filters were subjected to autoradiography. An IRC1334 mutation (346 bp fragment) was identified in one allele of the subject (YY), but his brother (TS) and And his sister (SK) were not observed.
- FIG. 6 is a diagram showing the relationship between the onset of NI DDM and the mutation in a family with an IR A831 mutation.
- ACG Th r 831
- the IR A831 mutation was identified in one allele of the subject (Yh.T.), two older brothers (IT and Yo. ⁇ .) and one older sister ( ⁇ .0.). Arrows indicate the person, squares indicate males, and circles indicate females.
- the ones painted in black are NI DDM patients, the black and white patterns are shown by IGT (Impart glucosolerance), and the shaded ones represent the deceased.
- the IR A831 mutation in the exon 13 PCR fragment cleaves a specific site of the restriction enzyme Cf0I.
- the Cf0I digest (322 bp) of the mutant PCR fragment appears as bands of 102 bp and 220 bp, but this cleavage is inhibited in the wild type.
- FIG. 7 shows DNA encoding human insulin receptor (part 1).
- FIG. 8 shows DNA (part 2) encoding the human insulin receptor.
- FIG. 9 shows DNA encoding human insulin receptor (part 3).
- FIG. 10 shows DNA encoding the human insulin receptor (part 4).
- the mutated site (amino acid 831) of the mutated human insulin receptor of the present invention is indicated by a box.
- FIG. 11 shows DNA encoding the human insulin receptor (part 5).
- the mutation site (amino acid 1334) of the mutant human insulin receptor of the present invention is indicated by a box. Detailed description of the invention
- the nucleotide sequence encoding Thr at position 831 in human insulin receptor DNA is mutated to the nucleotide sequence encoding A1a and / or the nucleotide sequence encoding Tyr at position 1334 encodes Cys.
- the present invention provides a mutant human insulin receptor DNA mutated to a base sequence that changes, or the above mutant human insulin receptor DNA fragment containing the mutated portion.
- the insulin receptor specifically binds insulin and transmits that information into cells. It is a receptor of the biological membrane that reaches, consisting of two alpha chains (735 residues, molecular weight 84,214) and two / 3 chains (620 residues, molecular weight 69,700).
- the gene for the insulin receptor consists of 22 exons, 11 of which encode the alpha subunit and the other 1 iexon encodes the subunit (S. Seino et al., Proc. Natl. Acad. Sci. USA 86: 114-118, 1989).
- the nucleotide sequence of the human insulin receptor and the corresponding amino acid sequence are shown in SEQ ID NO: 1 in the sequence listing.
- the mutated human insulin receptor DNA of the present invention is characterized in that the nucleotide sequence ( ⁇ CG) encoding Thr at position 831 of subunit exon 13 is mutated to a nucleotide sequence (CG) encoding Ala.
- a mutant human insulin receptor DNA in which the nucleotide sequence (T ⁇ C) encoding Tyr at position 1334 of Subunit exon 22 has been mutated to a nucleotide sequence (T ⁇ C) encoding Cys, and the mutated portion thereof And a fragment of the mutant human insulin receptor DNA.
- the mutated site of the mutant human insulin receptor of the present invention is shown in the sequences shown in FIGS. 7 to 11 (same as the sequence of SEQ ID NO: 1 in the sequence listing) in a box.
- the mutant human insulin receptor DNA of the present invention is obtained by amplifying all 22 exon genes of the insulin receptor of 51 typical Japanese patients with NID DM by PCR (polymerase chain reaction) and inserting them into the pUC19 vector. Later, it was obtained by determining the DNA base sequence. As a result of DNA sequence analysis, the thr at position 831 of yS subunit exon 13 of one parental gene was mutated to A1a in three cases, and the 1334th position of subunit exon 22 of subunit exon 22 of one parental gene. One case was found in which Ding r was mutated to Cys.
- the mutated human insulin receptor DNA of the present invention constitutes a qualitative mutation of at least about 6% of the NIDDM onset gene, which is a significant value that enables the application of NIDDM to genetic diagnosis.
- NIDDM neurotrophic factor
- mutant human insulin receptor DNA containing the mutant portion of the present invention and a fragment of the mutant human insulin receptor DNA, but also a DNA complementary thereto or a fragment thereof is useful as a diagnostic probe. is there. Therefore,
- the nucleotide sequence (TAC) encoding the Tyr at position 1334 of subunit exon 22 is a nucleotide sequence encoding Cys.
- TGC human insulin receptor DNA fragment mutated to stem ij
- the present invention is also useful as a cleavage probe for NIDM in the present invention.
- DNA fragments for a diagnostic probe generally have a length of up to about 100 bases containing the above-mentioned mutated portion, preferably 10 to 50 bases containing the mutated portion, more preferably 10 to 30 bases containing the mutated portion. Consists of an array.
- the DNA or DNA fragment of the present invention can be synthesized, for example, according to the base sequence of the present invention using a DNA automatic synthesizer utilizing a solid phase method on a support such as silica.
- the genetic diagnosis according to the invention of the present application is not limited to any particular method, and various methods may be widely adopted as long as the specific mutations identified and characterized by the present invention are detected. Can be. Since the gene mutation to be detected is clarified and identified by the present application, it will be easy for those skilled in the art to appropriately employ a method for the detection according to the present disclosure.
- such a method can be performed by analyzing the base sequence at the above-mentioned specified position, and naturally includes this.
- the Southern hybridization method and the dot hybridization method may be options.
- PCR Polymerase chain reaction; one RFLP method (Restriction fragment length polymor phism)
- PCR Single-chain higher-order structural polymorphism analysis method (Orita, M., Iwahana, H. , Kanazawa, ⁇ ., Hayashi, K. and Sekiya, T., Proc. Natl. Acad.
- PCR-SSO method Specific sequence oligonucleotide: P CR—specific sequence oligonucleotide method
- PCR—allele specific oligomer method using SSO and dot hybridization method allele specific oligomer: AS 0; Saiki, RK, Bugawan, TL, Horn, GT, Mullis, KB and Erlich, HA, Nature, 324: 163-166, 1986, etc.
- other methods that combine PCR with DNA amplification methods are simple and easy to use with small amounts of DNA samples. And high sensitivity and accuracy Detection is possible and can be preferably exemplified.
- the employment of the RFLP method and / or the allele-specific hybridization method can be preferably exemplified due to its simplicity.
- the detection method will be described in more detail by taking such a detection method as an example.
- Various operations that can be employed in the detection method of the present invention for example, chemical synthesis of part of DNA, enzymatic treatment for DNA cleavage, deletion, addition or binding, isolation, purification, replication of DNA, Selection and the like can be performed in accordance with the usual methods [Molecular Genetics Experimental Method, published by Kyoritsu Publishing Co., Ltd. in 1983; PCR technology, Takara Shuzo Co., Ltd. published in 1990, etc.].
- DNA isolation and purification can be performed according to agarose gel electrophoresis, and the DNA sequence can be determined, for example, by the dideoxy method (Sanger, F., Nicklen, S. and Coulson, AR, Proc. Natl. Acad. Sci.
- the determination of the DNA base sequence can be easily performed by using a commercially available sequence kit or the like.
- the PCR method for amplification of a specific region of DNA is also a conventional method (for example, Saiki, RK, Scharf, S., Faloona, FA, Mullis, KB, Horn, GT, Erlich, HA and Arnheim, N., Science, 230: 1350-1354, 1985, etc.).
- Saiki, RK, Scharf, S., Faloona, FA, Mullis, KB, Horn, GT, Erlich, HA and Arnheim, N., Science, 230: 1350-1354, 1985, etc. These various basic operations are also employed, for example, in various documents cited in the present application, and are referred to together with examples described later.
- the genomic DNA to be measured is a human-derived sample, and any sample containing the same can be used without particular limitation.
- Genomic DNA can be extracted, purified, and prepared from these samples by a conventional method.
- the DNA region containing the mutation site according to the present invention can be amplified to obtain a large and concentrated sample.
- This can be carried out, for example, according to the PCR method, and is carried out by employing a primer appropriately selected so as to specifically widen only the region containing the mutation site of exon 13 or 22.
- the setting of such a primer may be carried out according to a conventional method, and there is no limitation on the base length of the region to be amplified, and it can be usually from 100 bp to 500 bp. Suitable examples of such primer settings include, for example, flanking intronic sequences. primers (Seino, S., Seino, M.
- a sense primer 5'-CACTGACC TCATGCGCATGTGCTGG-3 'and an antisense primer: 5'-ATTGGACCGAGGCAAGGTCAGAAT-3' are employed.
- Such a primer set provides the desired regions as amplified DNA fragments of 322 bp (exon 13) and 346 bp (exon 22), respectively.
- the mutation specific to the present invention contained in the region can be detected and confirmed.
- the detection of the mutation relating to exon 13 is performed according to the RFLP method.
- Mutation of A la 831 (GCG) results in a specific cleavage site for the restriction enzyme CfoI. Therefore, the PCR amplification product of exon 13 having the mutation (322 bp) gave two fragments of 102 bp and 220 bp by subjecting it to Cf0I digestion. This cleavage does not occur in the wild type (322 bp). The generated fragment is confirmed as a specific band according to a conventional method.
- an allele-specific hybridization method using the above-described DNA fragment for a diagnostic probe is employed. This can be performed according to a conventional method as long as the specific mutation is detected by using the specific diagnostic probe described above.
- the conditions adopted in Examples described below are as follows.
- Hybridization of the PCR amplified product of exon 22 (346 bp) transferred to a nitrocellulose filter was performed using a probe solution (6 x SSC, 10 x Denhardt's solution, 1% SDS, 1 mg Zm1 salmon sperm) DNA and 32 P-labeled probe) at 30 ° C. overnight.
- the hybridized filters were washed twice (in 6 X SSC with 0.1% SDS, 54 ° C, 20 minutes each) and then subjected to conventional autoradiography.
- the wild-type (Ty r 1334: TAC) IR can distinguish from C 1334 mutation (Cy s 1334: TGC) Japanese A unique probe is the 32 P-labeled probe: 5'-ATGTGTGTG C ⁇ AAGGG ATGT-3 '.
- the present invention also provides such a non-insulin-dependent diabetes diagnostic agent.
- the diagnostic agent contains a specific reagent as an essential component according to the method for detecting the presence of the mutation according to the present invention.
- a specific reagent is appropriately set in accordance with the detection method to be employed.
- the specific reagent is used as a means for specifically detecting the mutation according to the present invention such as the DNA fragment for a diagnostic probe and a specific restriction enzyme. Characterized as a required reagent.
- reagents for specifically PCR-amplifying the region related to the mutation of the present invention for example, a primer set for the PCR are not considered to be essential components of the diagnostic agent of the present invention. Similar to the reagents for bridging, it can be included in the diagnostic agent of the present invention.
- Example 1 Isolation of human chromosome DNA
- the insulin receptor gene was amplified by the PCR method using the primer DNA capable of amplifying all 22 exons of one gene.
- dNTPs mix (2.5mM) 8.0 ⁇ 200 iiM
- the PCR-amplified DN ⁇ fragment obtained in this manner was purified by agarose gel electrophoresis, ligated to the H inc II site of pUC19 treated with allyl phosphatase, and the recombinant was introduced into E. coli. . Twelve colonies were picked out from hundreds of ampicillin-resistant colonies, and it was confirmed for each exon that 50% or more of the colonies contained the PCR DNA fragment. The remaining hundreds of colonies were all scraped off from the agar medium, grown in a liquid medium, and the crude plasmid DNA was separated by an alkali method, and RNA was removed by a polyethylene glycol precipitation method. When a large number of E.
- V a 1 830 -Th r 831 H is 832
- V a 1830 18 1 a 831 -H is 832
- FIG. 1 shows a photograph of electrophoresis showing a part of the DNA base sequence of the insulin receptor; S subunit exon 13 of five NID DM patients including the NID DM patient having this mutation.
- the third patient lane was found to der heterozygote having both sequences of Th r 831 (ACG) and A la 831 (GCG). There were two other patients with the same sequence.
- Another example of a mutation involving amino acid substitution had the following mutation in which the amino acid Tyr at position 1334 of insulin subunit 3 subnexon 22 was mutated to Cys.
- FIG. 2 shows a photograph of electrophoresis showing a part of the DN ⁇ nucleotide sequence of the insulin receptor ⁇ -subunit texon 22 of eight NI DDM patients including the NI DDM patient having this mutation. 7 th patients lane has been found to be heterozygote with both sequences of Ty r 1334 (TAC) and C ys 13 34 (TGC).
- TAC Ty r 1334
- TGC C ys 13 34
- IR A831 (those Th r 831 is mutated to A 1 a 831) and IR c is3 4 (Ty r 1334 is Cy s1334 ).
- CHO cells were transfused with SRa IR WT , SR «IR A831 or SRC1334 (10 zg each), and pSV2-neo (1 // g) by the calcium phosphate precipitation method.
- SRa IR WT SR «IR A831 or SRC1334 (10 zg each)
- pSV2-neo (1 // g) by the calcium phosphate precipitation method.
- G418 Sigma
- cells expressing the human insulin receptor were transformed by the method described in the literature (H. Hayas hi et al., Biochem. J. 280: 769-775, 1991).
- 125 I-labeled insulin binding was calculated by Scatchard analysis (G. Scatchard, Ann. NY Acad. Sci. 51: 660-672, 1949).
- Insulin-stimulated receptor autophosphorylation of IR WT and IR A831 was performed in 96-well plates using the method described in the literature (H. Hayashi et al., Biochem. J. 280: 769-775, 1991). .
- the incorporation of 32 P into the receptor 3—subunit was detected by 6% SDS-PAGE and measured with a Bioimage analyzer BAS2000.
- PI 3- kinase a type p 85 Sabuyuni' bets, as well as wild-type or human E mutant insulin receptions evening - in order to express the (IR) transient, L ip 0 fectamine TM Reagent (B ethesda R esearch L aboratories) COS-7 cells were transfected with S p85a ( 1.5 ⁇ g and S Ra IR, SRIR A831 or SRa IR C1334 ( 1.5 g each) using COS- 7 cells.
- a cell lysate was prepared and incubated with an anti-IR antibody 1G2 recognizing an IR subunit, or an anti- ⁇ 85 ⁇ antibody from Escherichia coli and protein G-Sepharose.
- the immunoprecipitate was electrophoresed on a 6% SDS-PAGE, and the immunoblot was performed using an anti-phosphotyrosine antibody (PY20) or an anti-insulin receptor-monoclonal antibody that recognizes the ⁇ -subunit of IR 3-11. .
- FIG. 3A shows a Scatchard yard analysis of IR WT (clone 12, 21 ) and IR A831 (clone 10, 17).
- the number of insulin binding sites in each clone was determined.
- the number of high affinity binding sites, respectively for 1 cells per 1.1 IR WT and IR A831, 1. 5 xl 0 6 and 0.5, a 0. 9 X 10 6, dissociation constant (Kd) is 2 8, 3.5 nM and 1.7, 2.7 nM.
- the autophosphorylation activity of these receptors was measured by the test method described in (3) above. The results obtained are shown in FIG. 3B. As is clear from the figure, the receptor autophosphorylation activity increased with the number of receptors. 50% maximum (11 & 1 f — ma xi In ma 1) stimulation IR WT and IR A831 respectively, 5. 5, 5. 4x 10 - 10 M and 5.0, a 4. 9 x 10- 1 ⁇ ⁇ , prominent among the wild-type and artificial mutant There was no significant difference.
- Insulin binding and receptor autophosphorylation were then tested using COS cells transiently expressing IR WT and IR A831 to eliminate differences between CHO clones stably expressing each IR. As a result, no remarkable difference was observed in the insulin binding affinity and the receptor kinase activity between the wild type and the artificial mutant receptor. No significant differences were observed between wild-type and artificially mutated receptors in receptor processing, internalization, degradation, insulin-stimulated glucose uptake, glycogen synthesis, and DNA synthesis. Therefore, no significant impairment of receptor function was found for the A831 mutation.
- IR autophosphorylated Tyr 1334 is located in a putative binding motif (Y (P) XXM) with the SH2 domain of the 85-kDa regulatory subunit of phosphatidylinositol 3-kinase (p85).
- Y (P) XXM putative binding motif
- Autophosphorylation to PI 3-kinase in response to insulin This mechanism may be an alternative pathway for activation of PI 3-kinase by IRS-1, as IR binding activates this enzyme. Absent.
- PI 3-kinase is a glucose transporter type 4
- the p85 subunit of PI 3-kinase was transiently expressed in COS cells using IR WT , IR A831 and IR C1334 .
- cell lysates were precipitated with either anti-IR3 antibody ( ⁇ IR ⁇ ) or anti-p85 antibody ( ⁇ 85) (Figure 4).
- Immunoprecipitates were tested by Immunobout using an anti-phosphotyrosine antibody ( ⁇ ) (Fig. 4 ⁇ ) or an anti-IR submit antibody (Fig. 4B).
- insulin treatment was used. by, PI 3 -.
- IR C1334 showed IR in CHO cells stably expressing IR against autophosphorylation site and endogenous substrate poly ( Glu , Tyr ) 4: 1.
- Anti-phosphotyrosine antibodies appear to recognize the site of autophosphorylated Y1334 better than the other sites of autophosphorylation of I, as they show almost the same degree of tyrosine kinase activity as WT and IR A831 .
- Anti-p85 antibody co-precipitated IR WT and IR A831 , but not IRdsa (FIG. 4B). This suggests that the major binding sites of the IR is Y 1334 residues to Ikoto, and p 85 a bonded IR C1334 is a p 85.
- Example 5 Statistical processing
- Non-diabetic patients are not diagnosed by performing tests such as OGTT, and their parents, siblings, and both grandparents, including themselves, have no symptoms of diabetes and have been diagnosed with diabetes and visited hospitals, etc. He was treated as non-diabetic.
- IR A831 A family with IR A831 to test the association between IR A831 and NI DDM was analyzed. The father was dead and could not be confirmed for the mutation, but was known to have NI DDM. The mother was too old to be tested. Also, of his four siblings, including himself, three were NID DMs and one was an intermediate between normal and diabetic IGT (Imparedglucosetoler an ce), and all four were 11 ⁇ 831 116 1 6 1 * 0 272 0 te. (See Figure 6). The detection of IR A831 was performed by the above-described method of generating a specific cleavage site for the restriction enzyme Cf0I (FIG. 6B). Family marriage was excluded. The allele of IR A831 appeared to be from the father. This pedigree analysis strongly suggests that the heterozygous gous mutation in IR A831 is responsible for the development of NI DDM.
- Sequence length 4723 (139-4287) Sequence type: nucleic acid
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Description
Claims
Priority Applications (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP95918175A EP0803570A4 (en) | 1994-05-12 | 1995-05-12 | MUTANT DNA RECEPTOR OF HUMAN INSULIN |
US08/737,715 US5958685A (en) | 1994-05-12 | 1995-05-12 | Mutant human insulin receptor DNA |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
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JP13482794 | 1994-05-12 | ||
JP6/134827 | 1994-05-12 |
Publications (1)
Publication Number | Publication Date |
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WO1995031542A1 true WO1995031542A1 (fr) | 1995-11-23 |
Family
ID=15137403
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
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PCT/JP1995/000906 WO1995031542A1 (fr) | 1994-05-12 | 1995-05-12 | Adn mutant recepteur de l'insuline humaine |
Country Status (4)
Country | Link |
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US (1) | US5958685A (ja) |
EP (1) | EP0803570A4 (ja) |
CA (1) | CA2190120A1 (ja) |
WO (1) | WO1995031542A1 (ja) |
Families Citing this family (2)
Publication number | Priority date | Publication date | Assignee | Title |
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GB9924717D0 (en) * | 1999-10-19 | 1999-12-22 | Glaxo Group Ltd | Diagnostic |
EP3441471A1 (en) * | 2017-08-08 | 2019-02-13 | CEVEC Pharmaceuticals GmbH | Use of constitutively active variants of growth factor receptors as selection makers for the generation of stable producer cell lines |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
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US5260200A (en) * | 1991-01-18 | 1993-11-09 | Joslin Diabetes Center, Inc. | Isolated DNA encoding an insulin receptor substrate |
-
1995
- 1995-05-12 EP EP95918175A patent/EP0803570A4/en not_active Withdrawn
- 1995-05-12 US US08/737,715 patent/US5958685A/en not_active Expired - Fee Related
- 1995-05-12 CA CA002190120A patent/CA2190120A1/en not_active Abandoned
- 1995-05-12 WO PCT/JP1995/000906 patent/WO1995031542A1/ja not_active Application Discontinuation
Non-Patent Citations (5)
Title |
---|
DIABETES, (1992), Vol. 41, No. 4, COCOZZA S. et al., "NIDDM Associated with Mutation in Tyrosine Kinase Domain of Insulin Receptor Gene", pages 521-6. * |
DIABETES, (1993), Vol. 42, No. 3, ELBEIN S.C. et al., "Methionine For Valine Substitution in Exon 17 of the Insulin Receptor Gene in a Pedigree With Familial NIDDM", pages 429-34. * |
DIABETOLOGIA, (1992), Vol. 35, No. 3, KIM H. et al., "Detection of Mutations in the Insulin Receptor Gene in Patients With Insulin Resistance by Analysis of Single-Stranded Conformational Polymorphisms", pages 261-6. * |
DIABETOLOGIA, (1992), Vol. 35, No. 5, O'RAHILLY S. et al., "Insulin Receptor and Insulin-Responsive Glucose Transporter (GLUT4) Mutations and Polymorphisms in a Welsh Type 2 (Hon-Insulin-Dependent) Diabetic Population", pages 486-9. * |
See also references of EP0803570A4 * |
Also Published As
Publication number | Publication date |
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EP0803570A4 (en) | 1998-05-20 |
US5958685A (en) | 1999-09-28 |
CA2190120A1 (en) | 1995-11-23 |
EP0803570A1 (en) | 1997-10-29 |
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