JPH10330285A - Hypoglycemic agent - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain hypoglyceoic agent capable of controlling a blood glucose level of a diabetes patient, especially useful for insulin-independent type II by including a specific growth factor as an active ingredient. SOLUTION: This hypoglycemic agent contains a fibroblast growth factor 10 (FGF-10) or a keratinocyte growth factor 2 (KGF-2) as an active ingredient. The growth factor is preferably a polypeptide having an amino acid sequence of the formula, or an additionally, deficiently or substitutively modified polypeptide thereof. The growth factor is obtained, for example, by extracting a host obtained by transducing a recombinant DNA. The hypoglycemic agent is capable of gradually improving hyperglycemic state without causing sudden hypoglycemia such as insulin.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a hypoglycemic agent, and more particularly to a hypoglycemic agent for treating diabetes, comprising fibroblast growth factor 10 (FGF-10) as an active ingredient.

[0002]

2. Description of the Related Art In recent years, the number of patients with diabetes and diabetic complications has been increasing along with the change to a Western-style diet and an increase in the tendency to lack exercise due to the improvement in living standards.

[0003] Generally, diabetes is dependent on insulin (I).
Type, IDDM) and insulin-independent (type II, NID)
DM), and more than 90% of diabetic patients are the latter. Insulin injection for treatment of IDDM, NIDD
For the treatment of M, sulfonylurea and biguanide oral diabetes drugs are selected together with exercise therapy and diet therapy ["Today's treatment guideline TODAY'S THERAPY".
1993] Shigeaki Hinohara, Masakazu Abe, Medical Shoin, 49
4 to 498]. However, fundamental pharmacotherapy for diabetes has not yet been established, and visual impairment, peripheral neuropathy, delayed wound healing,
At present, various complications such as ulceration and obesity are caused.

On the other hand, fibroblast growth factor 10 (FGF-
10) is a growth factor whose expression and physiological activity have been confirmed by recombinant production for the first time by the group of Ito et al., Kyoto University [Japanese Patent Application No. 8-214378]. Structurally FG
Belonging to the F family, especially keratinocyte growth factor: K
Fibroblast growth factor 7 (FG), also called GF-1
F-7) has about 60% amino acid homology. Also,
At about the same time, Gruber, JR, et al., Also code for KGF, which encodes the same amino acid sequence as FGF-10.
GF-2 gene has been discovered [Wo96 / 2542]
2: Human Genome Science In
c. ]. Although FGF-10 (KGF-2) has been confirmed to have a proliferative action on epithelial cells and bone cells, no action has yet been reported on the action of lowering the blood glucose level of a living organism in a hyperglycemic state.

[0005]

An object of the present invention is to provide a novel antihypertensive agent capable of safely controlling the blood sugar level of a diabetic patient. In particular, it provides an antihypertensive agent effective in type II diabetes, which is insulin-independent.

[0006]

Means for Solving the Problems The present inventors have studied the effects of peripherally administered neurotrophic factors on various diabetes model animals. Certain spontaneously hyperglycemic mice (ob / ob
Mouse) [Herberg and Coleman, Metabolism, 26, 59-99 (1977): Herber
g, L., and DLColeman. 1977. Laboratory animals
exhibiting obesity and diabetes syndromes.Metabol
ism 26: 59-99.], it was found that administration of FGF-10 could lower blood glucose levels. Based on this finding, as a result of further studies, the present invention has been completed.

That is, the present invention relates to the following medicaments. (1) A hypoglycemic agent containing fibroblast growth factor 10 (FGF-10) or keratinocyte growth factor 2 (KGF-2) as an active ingredient. (2) A hypoglycemic agent containing, as an active ingredient, a growth factor which is a polypeptide consisting of the amino acid sequence of SEQ ID NO: 1 or an addition, deletion or substitution variant thereof. (3) The hypoglycemic agent according to (1) or (2), wherein the growth factor is a recombinant protein produced by an E. coli host. (4) The hypoglycemic agent according to (1) to (3), which is used for treating diabetes. (5) The hypoglycemic agent according to (4), wherein the diabetes is non-insulin-dependent diabetes (type II diabetes, NIDDM).

Hereinafter, the present invention will be described in detail. As used herein, "fibroblast growth factor 10 (FGF-1
0) or keratinocyte growth factor 2 (KGF-
2) "means the F which was discovered by Ito et al. In 1996.
Cell growth factor of GF family [Japanese Patent Application No. 8-21437]
8] having the amino acid sequence of SEQ ID NO: 1
It refers to a protein having a proliferative action such as cells (cultured cells of an epithelial cell line) and promoting rat bone formation. In the following description, the term "FGF-10" is used to refer generally to a variant of FGF-10, that is, an addition, deletion, or substitution variant of a polypeptide consisting of the amino acid sequence of SEQ ID NO: 1. A method for preparing a typical variant and a method for measuring the activity are described in Japanese Patent Application No. 8-214378.

[0009] Although natural FGF-10 is a protein having a sugar chain, it is included in the concept of FGF-10 as long as it has the same kind of cell growth activity regardless of the presence or absence of the sugar chain. The FGF-10 mature protein includes (1) Leucine at position 40 (Leu40) of SEQ ID NO: 1, and serine at position 208 (Ser2).
08) 169 amino acid protein ending in
(2) A protein of 140 amino acids starting from serine 69 (Ser69) and ending at serine 208 (Ser208) is currently known.
10 is not limited to these two mature proteins.

[0010] The term "hypoglycemic agent" refers to a medicament which is administered to a patient in a pathologically hyperglycemic state to lower the blood glucose level.
“Diabetes” is a disease in which the blood sugar level is normally controlled to about 100 to 120 mg / dl and abnormally rises. According to current diagnostic criteria, the blood glucose level is
After 2 hours of g loading, 200 mg / dl or more and fasting blood glucose become 140 mg / dl or more. Generally, diabetes
Insulin dependence (type I, IDDM) with reduced insulin producing cells and insulin independent (type II, NID) allegedly caused by decreased insulin sensitivity
DM), and more than 90% of diabetic patients are the latter. The former is a young-onset type, in which insulin-producing cells are destroyed by autoimmunity, and the amount of insulin becomes insufficient. There is a strong tendency for ketosis, and continuous insulin injection is essential for life support. "Non-insulin dependent diabetes mellitus (type II diabetes mellitus, NIDDM)"
Is a type of diabetes that mainly develops after middle age, and is caused by a decrease in the sensitivity of somatic cells to insulin. Blood insulin levels are rather high. Oral hypoglycemic agents are more commonly used than insulin injections due to weak and slow progression of ketosis, but hyperglycemia indirectly causes various complications. In the present invention, in the present specification, a pathological hyperglycemic state (steroid diabetes), which occurs when a steroid drug such as glucocorticoid is continuously administered, and a hyperglycemic state in Cushing Syndrome and acromegaly, It is included in type II diabetes in the sense that it is normal or hyperinsulinic diabetes.

(Production) FG used in the hypoglycemic agent of the present invention
F-10 can be purified and used in the present invention, regardless of whether it is a natural extract or a genetically modified product, as long as it exhibits an intrinsic physiological activity. Methods for producing FGF-10 include (1) extraction from FGF-10 producing tissue, and (2) FGF-10 production.
Culture and extraction of GF-10 producing cells (primary cultured cells or cell lines), (3) culture and extraction of host cells into which recombinant DNA has been introduced, and the like can be considered.
(3) Extraction from a host into which recombinant DNA has been introduced is suitable for mass production. A method for producing FGF-10 by a recombinant technique is briefly described below.
14378.

(Recombination step) D represented by SEQ ID NO: 2
The FGF-10 cDNA containing the NA sequence is incorporated into an expression vector. As the vector, a plasmid or a phage which can be propagated in a suitable host such as Escherichia coli, Bacillus subtilis, yeast, and animal insect cells is selected.
BR322, pBR325 [Gene 4 vol. 1
21 (1978)], pUB110 derived from Bacillus subtilis [Biochemical Biophysical Research Communication (Biochem. Biophys. Res.
Commun. 112, 678 (1983)],
PCDM8 suitable for COS cells and the like. cDN
As a method of incorporating A into a plasmid, a common method is described in T. Maniatis et al.
Cloning (Molecular clonin
g), Cold spring harbor lab.
239 (1982).

(Host) The host is not particularly limited as long as it is transformed by introduction of a vector and can produce FGF-10 or a cultured cell. As bacteria, Escherichia coli,
Examples of yeast such as Bacillus subtilis (Bacillus) include Saccharomyces, Torula and Pichia, and animal cells include C.
OS cells, CHO cells, NSO cells and the like are typical examples.
In addition to cultured insect cells, fungi, plant cells, single cell systems,
Insects, mammals, and plants into which the target protein gene has been incorporated are also included in the category of the host.

(Measurement of activity) From the transformant, a known method such as a colony hybridization method [Gene, 10, 63 (1980)] and a DNA sequencing method [Proceedings of
National Academy of Sciences (Proc.
Natl. Acad. Sci. USA, vol. 74, p. 560 (1977)]. Alternatively, clones can be transiently expressed in COS cells and cloned by evaluating the physiological activity of the culture supernatant. The physiological activity of the expressed FGF-10 protein can be evaluated by measuring the growth promoting action of epithelial cells such as FRSK cells.

(Purification) FG produced by recombinant technology
The F-10 protein can be purified by a purification method commonly used in the field of biochemistry. Ion exchange chromatography,
Gel filtration, reverse phase HPLC, ammonium sulfate precipitation, ultrafiltration, SDS
-PAGE etc. are used in an appropriate combination.
In the case of Fs, affinity chromatography and antibody column chromatography using a ligand such as heparin are particularly suitable for large-scale purification. Antibodies to the FGF-10 protein can be prepared by a method known per se for both polyclonal and monoclonal antibodies. FGF-10-specific antibodies can be used not only for antibody columns, but also for ELISA.
And the like.

(Preparation) Preparations include injections, oral preparations,
Both liquid preparations and freeze-dried preparations can be used, but injection preparations for subcutaneous administration are particularly preferred. These parenteral preparations can use stabilizers and carriers known in the art, and are preferably used as isotonic solutions when used.
As the pharmaceutical carrier, for example, plasma-derived proteins such as albumin, amino acids such as glycine, and sugars such as mannitol can be used, and they are usually used in freeze-dried preparations for subcutaneous or intramuscular administration. When used as a water-soluble preparation or a lyophilized preparation, it is preferable to add a surfactant such as Tween 80 to prevent aggregation. When a long-term effect is required, the preparation can be prepared using a known protein release-release preparation carrier.

(Method of Use) When the main component of the hypoglycemic agent of the present invention is FGF-10, usually 0.5 μg to 5 mg per kilogram of an adult is administered intravenously, subcutaneously or intramuscularly. The frequency of administration may be appropriately adjusted depending on the dose, administration route and patient's symptoms, but administration may be performed once a month to three times a day, and is generally performed 1 to 5 times a week for several weeks. Drug treatment is performed. With this treatment, the blood sugar level is slowly reduced, and a moderate blood sugar level lower than that of a conventional antihypertensive agent is obtained and stabilized. Although the hypoglycemic effect of FGF-10 cannot be explained from conventional findings, it is considered that it is at least through a new pharmacological mechanism completely different from that of insulin or IGF.

(Toxicity) Normal mice (C57BL / 6N,
Male, 5 weeks old: 1 week in Charles River Japan, and obese diabetic mice (C57BL / 6J-Lep <
ob>, female, 6 weeks old: Charles River Japan, Ja
ckson laboratory) up to 5 weeks for 2 weeks
mg / kg of FGF-10 was administered intraperitoneally or subcutaneously, but there was no weight loss or death. Generally, toxicity is considered low.

[0019]

Industrial Applicability The hypoglycemic agent of the present invention gradually improves hyperglycemia without causing rapid hypoglycemia unlike insulin.

[0020]

The present invention will be described below with reference to examples. (Expression and purification of FGF-10) A DNA fragment (SEQ ID NO: 2) corresponding to the structural gene of human FGF-10,
Escherichia coli expression vector pET11c (Stratagene) is digested with NdeI and BamHI, linearized by agarose gel electrophoresis, ligated, and cloned by transforming Escherichia coli JM109. did. From these, the plasmid into which FGF-10 cDNA was inserted in the correct direction was isolated, the nucleotide sequence was confirmed, and pET-hFG was confirmed.
F-10 was obtained. Escherichia coli BL21 (DE
3) was transformed. One of the obtained recombinant clones was designated as BL21 (DE3) / pET-hFGF-10, and used to produce and produce human FGF-10.

(Culture) BL21 (DE3) / pET-h
L containing FGF-10 at 100 μg / ml ampicillin
Four cells inoculated in 10 ml of B medium were prepared and pre-cultured at 37 ° C. overnight. The next day, the total amount was 100 μg /
Inoculate 4x500ml TB medium containing 37ml at 37 ℃
With shaking. When OD600 reached 0.8, IPTG was added to a final concentration of 1 mM, the culture temperature was lowered to 28 ° C, and the culture was continued for another 6 hours.

(Extraction and Purification) The culture was centrifuged, and the obtained cells were cultured in 50 mM Tris-HCl, pH 8.0.
The cells were washed once, and suspended in 50 mM Tris-HCl, pH 8.0 containing 1 mM EDTA, 2 μg / ml leupeptin, 2 μg / ml pepstatin, and 1 mM PMSF.
The cells are disrupted by ultrasonic disruption, and Beckman J2-21
The supernatant was collected by centrifuging at 15,000 rpm for 1 hour using a JA-20 rotor in an M / E high-speed cooling centrifuge. HiTrap Heparin (5m
1, Pharmacia) in 50 mM Tris-HCl, pH
The mixture was equilibrated with 8.0, and the supernatant of the disrupted cells prepared above was applied. Subsequently, 50 mM Tris-HCl, pH 8.0.
After washing until the eluate A260 returned to the baseline, the protein was eluted by continuously increasing the NaCl concentration gradient to 3M. An approximately 19 kDa protein corresponding to recombinant human FGF-10 was eluted at approximately 1.2 M NaCl. The flow rate was 2 ml / min.

(Formulation Example) Among the FGF-10 preparations of the present invention, a typical aqueous / lyophilized preparation for subcutaneous administration can be produced as follows. (1) To 1 mg of purified recombinant FGF-10, 0.34 mg of glycine, 9 mg of mannitol, 0.2 mg of nonionic surfactant: polysorbate 80 were added, and the solution was added to 1 ml of phosphate buffer (pH 7.4, 5 mM). Dissolve and freeze-dry the solution. (2) 5 mg / ml of FGF-10 in a 10 mM phosphate buffer (pH 7.0) containing 150 mM sodium chloride and 0.01% Tween 80
To obtain an aqueous solution of FGF-10.
(3) 150 mM sodium chloride, 0.01% Tween
10 mM phosphate buffer (pH 7.0) containing n80
The FGF-10 was prepared so as to be 5 mg / ml.
Subsequently, mannitol is added to 10 mg / ml to obtain an FGF-10 aqueous solution. The vial is aseptically filled and freeze-dried according to a conventional method to obtain a freeze-dried FGF-10 preparation. Fill the vial with nitrogen and stopper.

(Pharmacological test) Obesity-type diabetes mellitus of 10 animals per group
bese (ob / ob) mouse [C57BL / 6J-L
ep <ob>, female, 6 weeks of age: Charles River Japan, Jackson laboratory], fed 1 and 5 mg / kg FGF-10 once a day for 2 weeks (subcutaneous administration). The control group (N = 5) received saline. In addition, a group of 5 normal mice [C57BL / 6N, male, 4 weeks old: Charles River Japan Co., Ltd.] were fed ad libitum and fed 1 and 5 mg / kg FG.
F-10 was administered once a day for one week (intraperitoneal administration). Blood was collected from the abdominal aorta 5 hours after the last administration in all groups, and serum was obtained. The blood glucose concentration was measured using an ultra-trace multipurpose automatic biochemical analyzer CHEM1 [Hexokinase method, Bayer].

Table 1 shows the results obtained using FGF-10.
And Table 2. By Student's t test, FG
The significant difference in the blood glucose level between the F-10 administration group and the control group was tested. Table 1: Blood glucose level of ob / ob mice Blood glucose level (mean ± SD) (mg / dl) Control group 453 ± 17 FGF-10 (1 mg / kg) administration group 307 ± 42 ** FGF-10 (5 mg / kg) ) Administration group 355 ± 27 ****: p <0.01

Table 2: Blood glucose in normal mice Blood glucose (mean ± SD) (mg / dl) Control group 215 ± 17 FGF-10 (1 mg / kg) administration group 224 ± 15 FGF-10 (5 mg / kg) Administration group 209 ± 32

As shown in Table 1, in the diabetic model group to which FGF-10 was administered, the blood glucose concentration was significantly reduced as compared with the control group, but the blood glucose level was not decreased in the normal mouse ( Table 2). Unlike insulin, which is liable to cause hypoglycemia, FGF-10 is expected to regulate blood sugar levels to the normal range.

SEQ ID NO: 1 Sequence length: 208 Sequence type: Amino acid Topology: Linear Sequence type: Peptide Origin Organism name: Human sequence Met Trp Lys Trp Ile Leu Thr His Cys Ala Ser Ala Phe Pro His Leu 1 5 10 15 Pro Gly Cys Cys Cys Cys Cys Phe Leu Leu Leu Phe Leu Val Ser Ser 20 25 30 Val Pro Val Thr Cys Gln Ala Leu Gly Gln Asp Met Val Ser Pro Glu 35 40 45 Ala Thr Asn Ser Ser Ser Ser Ser Ser Phe Ser Ser Pro Ser Ser Ala Gly 50 55 60 Arg His Val Arg Ser Tyr Asn His Leu Gln Gly Asp Val Arg Trp Arg 65 70 75 80 Lys Leu Phe Ser Phe Thr Lys Tyr Phe Leu Lys Ile Glu Lys Asn Gly 85 90 95 Lys Val Ser Gly Thr Lys Lys Glu Asn Cys Pro Tyr Ser Ile Leu Glu 100 105 110 Ile Thr Ser Val Glu Ile Gly Val Val Ala Val Lys Ala Ile Asn Ser 115 120 125 Asn Tyr Tyr Leu Ala Met Asn Lys Lys Gly Lys Leu Tyr Gly Ser Lys 130 135 140 Glu Phe Asn Asn Asp Cys Lys Leu Lys Glu Arg Ile Glu Glu Asn Gly 145 150 155 160 Tyr Asn Thr Tyr Ala Ser Phe Asn Trp Gln His Asn Gly Arg Gln Met 1 65 170 175 Tyr Val Ala Leu Asn Gly Lys Gly Ala Pro Arg Arg Gly Gln Lys Thr 180 185 190 Arg Arg Lys Asn Thr Ser Ala His Phe Leu Pro Met Val Val His Ser 195 200 205

SEQ ID NO: 2 Sequence length: 690 bp Sequence type: nucleic acid Number of strands: double-stranded Topology: linear Sequence type: cDNA origin Organism name: human sequence CTTCCAGTAT GTTCCTTCTG ATGAGACAAT TTCCAGTGCC GAGAGTTCCA GTACA ATG 58 TGG AAA TGG ATA CTG ACA CAT TGT GCC TCA GCC TTT CCC CAC CTG CCC 106 GGC TGC TGC TGC TGC TGC TTT TTG TTG CTG TTC TTG GTG TCT TCC GTC 154 CCT GTC ACC TGC CAA GCC CTT GGT CAG GAC ATG GTG TCA CCA GAG GCC 202 ACC AAC TCT TCT TCC TCC TCC TTC TCC TCT CCT TCC AGC GCG GGA AGG 250 CAT GTG CGG AGC TAC AAT CAC CTT CAA GGA GAT GTC CGC TGG AGA AAG 298 CTA TTC TCT TTC ACC AAG TAC TTT CTC AAG ATT GAG AAG AAG AGGAG 346 GTC AGC GGG ACC AAG AAG GAG AAC TGC CCG TAC AGC ATC CTG GAG ATA 394 ACA TCA GTA GAA ATC GGA GTT GTT GCC GTC AAA GCC ATT AAC AGC AAC 442 TAT TAC TTA GCC ATG AAC AAG AAG GGG AAA CTC TAT GGC TCA AAA GAA 490 TTT AAC AAT GAC TGT AAG CTG AAG GAG AGG ATA GAG GAA AAT GGA TAC 538 AAT ACC TAT GCA TCA TTT AAC TGG CAG CAT AAT G GG AGG CAA ATG TAT 586 GTG GCA TTG AAT GGA AAA GGA GCT CCA AGG AGA GGA CAG AAA ACA CGA 634 AGG AAA AAC ACC TCT GCT CAC TTT CTT CCA ATG GTG GTA CAC TCA TAGAG 684 GAAGGC 690

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[Procedure amendment]

[Submission date] August 18, 1998

[Procedure amendment 1]

[Document name to be amended] Statement

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[0028]

[Sequence list]  SEQ ID NO: 1 Sequence length: 208 Sequence type: Amino acid Topology: Linear Sequence type: Peptide Origin Organism name: Human sequence Met Trp Lys Trp Ile Leu Thr His Cys Ala Ser Ala Phe Pro His Leu 1 5 10 15 Pro Gly Cys Cys Cys Cys Cys Phe Leu Leu Leu Phe Leu Val Ser Ser 20 25 30 Val Pro Val Thr Cys Gln Ala Leu Gly Gln Asp Met Val Ser Pro Glu 35 40 45 Ala Thr Asn Ser Ser Ser Ser Ser Phe Ser Ser Pro Ser Ser Ala Gly 50 55 60 Arg His Val Arg Ser Tyr Asn His Leu Gln Gly Asp Val Arg Trp Arg 65 70 75 80

[Procedure amendment 1]

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[Correction method] Change

[Correction] Lys Leu Phe Ser Phe Thr Lys Tyr Phe Leu Lys Ile Glu Lys Asn Gly 85 90 95 Lys Val Ser Gly Thr Lys Lys Glu Asn Cys Pro Tyr Ser Ile Leu Glu 100 105 110 Ile Thr Ser Val Glu Ile Gly Val Val Ala Val Lys Ala Ile Asn Ser 115 120 125 Asn Tyr Tyr Leu Ala Met Asn Lys Lys Gly Lys Leu Tyr Gly Ser Lys 130 135 140 Glu Phe Asn Asn Asp Cys
Lys Leu Lys Glu Arg Ile Glu Glu Asn Gly 145 150 155 160 Tyr Asn Thr Tyr Ala Ser Phe Asn Trp Gln His Asn Gly Arg Gln Met 165 170 175 Tyr Val Ala Leu Asn Gly Lys Gly Ala Pro Arg Arg Gly Gln Lys Thr 180 185 190 Arg Arg Lys Asn Thr Ser Ala His Phe Leu Pro Met Val Val His Ser 195 200 205

[Procedure amendment 2]

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SEQ ID NO: 2 Sequence length: 690 bp Sequence type: nucleic acid Number of strands: double-stranded Topology: linear Sequence type: cDNA origin Organism name: human sequence CTTCCAGTAT GTTCCTTCTG ATGAGACAAT TTCCAGTGCC GAGAGTTCCA GTACA ATG 58 TGG AAA TGG ATA CTG ACA CAT TGT GCC TCA GCC TTT CCC CAC CTG CCC 106 GGC TGC TGC TGC TGC TGC TTT TTG TTG CTG TTC TTG GTG TCT TCC GTC 154 CCT GTC ACC TGC CAA GCC CTT GGT CAG GAC ATG GTG TCA CCA GAG GCC 202 ACC AAC TCT TCT TCC TCC TCC TTC TCC TCT CCT TCC AGC GCG GGA AGG 250

[Procedure amendment 2]

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[Correction method] Change

[Correction contents] CAT GTG CGG AGC TAC AAT CAC CTT CAA GGA GAT GTC CGC TGG AGA AAG 298 CTA TTC TCT TTC ACC AAG TAC TTT CTC AAG ATT GAG AAG AAC GGG AAG 346 GTC AGC GGG ACC AAG AAG GAG AAC TGC CCG TAC AGC ATC CTG GAG ATA 394 ACA TCA GTA GAA ATC GGA GTT GTT GCC GTC AAA GCC ATT AAC AGC AAC 442 TAT TAC TTA GCC ATG AAC AAG AAG GGG AAA CTC TAT GGC TCA AAA GAA 490

[Procedure amendment 2]

[Document name to be amended] Statement

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[Correction method] Change

[Correction contents] TTT AAC AAT GAC TGT AAG CTG AAG GAG AGG ATA GAG GAA AAT GGA TAC 538 AAT ACC TAT GCA TCA TTT AAC TGG CAG CAT AAT GGG AGG CAA ATG TAT 586 GTG GCA TTG AAT GGA AAA GGA GCT CCA AGG AGA GGA CAG AAA ACA CGA 634 AGG AAA AAC ACC TCT GCT CAC TTT CTT CCA ATG GTG GTA CAC TCA TAGAG 684 GAAGGC 690

Continued on the front page (51) Int.Cl. ⁶ Identification code FI C12R 1:19)

Claims (5)

[Claims] The present invention relates to fibroblast growth factor 10 (FGF-1).
0) or keratinocyte growth factor 2 (KGF-
A hypoglycemic agent containing 2) as an active ingredient. 2. A hypoglycemic agent comprising, as an active ingredient, a growth factor which is a polypeptide consisting of the amino acid sequence of SEQ ID NO: 1 or an addition, deletion or substitution variant thereof. 3. The hypoglycemic agent according to claim 1, wherein the growth factor is a recombinant protein produced by an E. coli host. 4. The hypoglycemic agent according to claim 1, which is used for treating diabetes. 5. The method according to claim 1, wherein the diabetes is non-insulin-dependent diabetes (I).
The hypoglycemic agent according to claim 4, which is type I diabetes mellitus (NIDDM).