JP2000319193A - Agent for prophylaxis and/or therapy of tissue disorder due to anticancer therapy or radioactive rays - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain the subject therapeutic agent useful for the prophylaxis and/or therapy of tissue disorder due to anticancer therapy or radioactive rays by including a heparin-binding secretory transforming factor (HST-1) DNA structure and a HST-1 protein (or its equivalent). SOLUTION: A propagation- and/or differentiation-stimulating agent for stem cells, productive cells, hair-mother cells, intestinal cells, villous tissue, mucous membrane, glomerulus endothelial cells and spleen cells is obtained by including (A) a heparin-binding secretory transforming factor (HST-1) DNA structure and (B) a HST-1 protein (or its equivalent). The above ingredients can usually be administered intravenously, and dosages and administration frequency usually are in the case of an adult (body weight 60 kg) as follows: about 0.0001-100 mg (preferably 0.0005-50 mg) ingredient A and about 0.0001-200 mg (preferably 0.0005-100 mg) ingredient B and; being administered respectively once or more times a day, or through continuous intravenous drip injection.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a heparin-binding secretory transforming factor, heparin-binding secretor.
y transforming factor (also called FGF-4; hereinafter, HS
The present invention relates to a prophylactic and / or therapeutic agent for tissue damage caused by anticancer therapy or radiation containing T-1) or HST-1 gene or the like.

[0002]

BACKGROUND OF THE INVENTION The HST-1 gene has been cloned as a transforming gene from human gastric cancer tissue for the first time by the present inventors and is located on chromosome 11q13 (Pro
c. Natl. Acad. Sci. USA, 83 , 3997-4001 (1986); ibi.
d., 84 , 2980-2984 (1987); Methods Enzymol., 198 , 1
24-138 (1991)). BFGF without signal sequence (FGF-2
Unlike aFGF (also called FGF-1)
Since ST-1 has a signal sequence, it immediately exits the cell after being made as a protein and can act on target cells efficiently (see “Experimental medicine bioscience / cell proliferation”). Factor Biology ”p.51-57,
Published by Sanshusha, 1992).

[0003] The HST-1 protein acts on megakaryoblasts in bone marrow to induce its differentiation and platelet production.
US 5,559,093 discloses that the HST-1 protein and its deletion proteins increase platelets and restore platelet counts decreased by thrombocytopenia, anticancer drugs or radiation. J. Clin. Invest., 96 , 1125-1130 (19
95) and Leukemia, 11 , 530-532 (1997) show that gene therapy using an adenovirus vector incorporating the HST-1 gene is applied to thrombocytopenia, a side effect induced by radiation and anticancer drugs. It states what you can do. However, HST-1 gene and HST-1 protein have the effect of promoting the proliferation and / or differentiation of stem cells, germ cells, hair cells, intestinal cells, villous tissue, mucous membranes, glomerular endothelial cells and spleen cells. Was not known.

[0004]

An object of the present invention is to provide an agent for preventing or treating tissue damage caused by anticancer therapy or radiation.

[0005]

Means for Solving the Problems The present inventors have found that the HST-1 gene has an effect of promoting the differentiation / proliferation of actively differentiated and / or proliferating tissues and cells,
The present invention has been completed.

That is, the present invention is as follows. [1] Proliferation and / or proliferation of stem cells, germ cells, hair cells, intestinal cells, villous tissue, mucous membranes, glomerular endothelial cells and spleen cells containing the HST-1 DNA construct, HST-1 protein or its equivalent Differentiation promoter. [2] An agent for preventing and / or treating tissue damage (excluding thrombocytopenia) due to anticancer therapy or radiation, comprising an HST-1 DNA construct, an HST-1 protein or an equivalent thereof. [3] The prophylactic and / or therapeutic agent according to [2], wherein the anticancer therapy is radiation therapy. [4] The damaged tissue is a tissue composed of hematopoietic stem cells, intestinal stem cells or neural stem cells [2] or [3].
The prophylactic and / or therapeutic agent as described above. [5] The prophylactic and / or therapeutic agent according to [2] or [3], wherein the damaged tissue is bone marrow or intestinal tract.

[6] Tissue disorders include leukopenia, anemia,
The prophylactic and / or therapeutic agent according to [2] or [3], which is erythrocytopenia or mucositis. [7] The prophylactic and / or therapeutic agent according to [2] or [3], wherein the tissue damage is bone marrow necrosis or a decrease in peripheral blood cells. [8] Tissue disorder is intestinal bleeding, reduction of villous tissue or 2
The prophylactic and / or therapeutic agent according to [2] or [3], which is a secondary infectious disorder. [9] The medicament according to any one of [1] to [8], which comprises an HST-1 DNA structure. [10] The medicament according to [9], wherein the HST-1 DNA construct is introduced into a vector. [11] The medicine according to [9], wherein the HST-1 DNA construct is introduced into an adenovirus vector or a plasmid vector.

[0008]

DETAILED DESCRIPTION OF THE INVENTION The HST-1 DNA construct refers to a construct containing HST-1 DNA or its equivalent, and additional sequences necessary to express the HST-1 protein or its equivalent. HST-1 DNA is defined as Proc. Natl. Acad. Sci. USA., 8
4 , DNA containing the HST-1 cDNA sequence described in 2980-2984 (1987)
Represents As an equivalent of HST-1 DNA, for example, "a protein consisting of an amino acid sequence in which one or more amino acids are substituted, deleted and / or added in the amino acid sequence of HST-1 protein, and the same as HST-1 protein DNA encoding an HST-1 DNA), a DNA that hybridizes with HST-1 DNA under stringent conditions, and whose translation product has a function similar to that of the HST-1 protein. Is mentioned. Examples of the additional sequence include a sequence derived from a eukaryote such as a promoter sequence and a polyA signal sequence of SV40. A promoter can be selected in consideration of expression efficiency and tissue specificity. As a promoter, for example, SRα
Promoter (Mol. Cell. Biol., 8 , 466-472 (198
8)), cytomegalovirus (CMV) promoter, LTR sequence promoter derived from Rous monkey virus, and the like. If high expression efficiency is not required, use MMTV
-LTR promoter, mouse metallothionein-1 (mMT-1)
Promoters and the like are useful.

The HST-1 protein is defined as Proc. Natl. Acad.
HST-1 protein described in Sci. USA., 84 , 2980-2984 (1987). The equivalent of the HST-1 protein includes, for example, a protein which is produced by expression of an equivalent of the HST-1 DNA and is different from the HST-1 protein. HST
-1 protein and the equivalent of HST-1 protein are HST-1
It may have at least one sugar chain, fatty acid chain or acetyl as long as it has the same activity as the protein. HST-
As an equivalent of one protein, for example, 27, 30, 31, 43, 46 from the N-terminal of the 206 amino acid sequence of the HST-1 protein
Or a protein in which 47 amino acids have been deleted (M
ol.Cell.Biol., 8 , 2933-2941 (1988); US 5,639,86
2). The activity of the HST-1 protein may be determined, for example, by promoting the DNA synthesis of mouse BALB / c3T3 cells by [ ³ H] -thymidine incorporation (Mo
l. Cell. Biol., 8 , 588-594 (1988)), promotion of vascular endothelial cell proliferation (J. Immun. Methods, 93 , 157 (1986)), angiogenesis in avian embryo chorioallantoic membrane (Dev. Biol. ., 41 , 391 (1974))
It can be checked by measuring.

[0010] Anticancer therapy includes chemotherapy and radiation therapy. Chemotherapeutic agents include, for example, alkylating agents (nitrogen mustard N-oxide,
Cyclophosphamide, melphalan, carbone, busulfan, nimustine, ranimustine, thiotepa, dacarbazine, procarbazine, etc., antimetabolites (fluorouracil, tegafur, cytarabine, ancitabine, gemcitabine, doxyfluridine, mercaptopurine, thionoisin, azathioprine, methotrexate, etc.) ,
Antibiotics (mitomycin, bleomycin, daunorubicin, doxorubicin, pirarubicin, aclarubicin, neocarzinostatin, actinomycin D, etc.),
Alkaloids (vinorelbine, paclitaxel, docetaxel, irinotecan, vincristine, vindesine,
Vinblastine, etoposide, etc.), hormonal agents (tamoxifen, etc.), platinum preparations (cisplatin, carboplatin, netaplatin, etc.), mitoxatron and the like.

[0011] Anticancer therapy usually causes damage to tissues and cells that are actively differentiated and / or proliferated. Examples of such tissues and cells include various stem cells (blood stem cells, myeloid stem cells, lymphocyte stem cells, intestinal stem cells,
Neural stem cells), germ cells, hair mother cells, intestinal cells, villous tissue, mucous membranes, glomerular endothelial cells, spleen cells and the like. Disorders of these tissues and cells include necrosis, atrophy and poor regeneration of tissues and organs including these tissues and cells, and typically include bone marrow necrosis, intestinal bleeding, villous destruction, Secondary infection due to the decrease is mentioned. And ultimately, death. In addition, since neural stem cells are present particularly in the hippocampal dentate region near the ventricle, nerve damage accompanying damage to this portion is caused. The necrosis or loss of these tissues or nerve cells leads to functional disorders such as movement disorders. The medicament of the present invention includes stem cells, germ cells, hair mother cells, intestinal cells, villous tissue, mucous membranes,
It can promote the proliferation and / or differentiation of glomerular endothelial cells and spleen cells. Therefore, it can be used as an agent for preventing and / or treating tissue damage due to anticancer therapy or radiation. Since the present medicament has a preventive effect for such disorders, the use of this medicament enables more effective anticancer therapy. This medicine is used for mucositis (intestinal mucositis, oral mucositis, etc.), leukopenia (neutropenia, eosinopenia, lymphopenia, etc.), anemia,
It is also effective for erythropenia. Further, the medicament is also effective as an adjuvant therapy for bone marrow transplantation therapy.

[0012] The HST-1 DNA construct can be applied as a gene therapy. Gene therapy can be broadly divided into in vivo and ex vivo methods (Nikkei Science, 1994 (4), 20-45, Monthly Pharmaceutical Affairs, 36 (1), 23-48 (19
94), Experimental Medicine Special Issue, 12 (15), 1298-1933 (1994), and references cited therein). That is, a method of directly administering an HST-1 DNA construct to a living body (in vivo method) and a method of removing specific cells (eg, bone marrow cells) from a living body and excluding HST-1 DN in vitro.
It can be applied by a method of introducing the A structure into the cell and returning the cell to the body (ex vivo method).

Examples of the in vivo method include a viral vector method, a liposome vector method, a fusion liposome vector method, and a lipofectin vector method (Nikkei Science, 1994 (4), 20-45, Monthly Pharmaceutical, 36 (1) ), twenty three-
48 (1994), Special Issue on Experimental Medicine, 12 (15), 1298-1933 (199)
4), Cardiovascular Research, 28 , 445 (1994), Scien
ce, 256 , 808 (1992), Gastroenterology, 106 , 1076
(1994), TIBTECH, 11 , 182, (1993), J. Biol. Chem.,
266 , 311 (1991), Nature Medicine, 1 , 583 (1995) and references therein). As a virus vector method, for example, an HST-1 DNA construct is converted into a DNA virus vector (adenovirus vector, adeno-associated virus (AAV) vector, herpes virus vector, vaccinia virus vector, poxvirus vector) or RNA
It can be carried out by incorporating it into a virus vector (poliovirus vector, Sindbis virus vector, retrovirus vector, Sendai virus vector, etc.). Adenovirus vectors such as adenovirus type 5 (ATCC No. VR-5) are particularly preferred because they can be made at high concentrations, are stable and easy to handle.

[0014] As the liposome vector in the liposome vector method, those produced from phospholipids can be used. Examples of the phospholipid include phosphatidylcholine (lecithin, lysolecithin, etc.), acidic phospholipids (phosphatidylserine, phosphatidylglycerol, phosphatidylinositol, phosphatidylic acid, etc.), and their acyl groups substituted with lauroyl group, myristoyl group, oleoyl group, etc. Phospholipids and sphingolipids (phosphatidylethanolamine, sphingomyelin, etc.) and the like. Neutral lipids such as cholesterol can be added to these phospholipids. The liposome can be produced from a natural material such as a lipid existing in a normal cell membrane by a generally known method. The liposome containing the HST-1 DNA structure can be produced, for example, by suspending a thin film of a purified phospholipid in a solution containing the HST-1 DNA structure and subjecting it to ultrasonic treatment or the like. The HST-1 DNA construct may take any form as long as it can be expressed in vivo. Preferred forms include in vivo stable forms such as plasmids. Examples of the fusion liposome in the fusion liposome vector method include a liposome fused with a virus, and preferably a membrane fusion liposome fused with a sender virus (HVJ) (Nikkei Science, 199).
4 (4), 32-38, J. Biol. Chem., 266 (6), 3361-3364 (19
91)). This membrane-fused liposome is prepared, for example, by mixing purified HVJ inactivated by ultraviolet irradiation or the like and a liposome suspension containing the HST-1 DNA structure, stirring gently, and then removing unbound HVJ. It can be prepared by removing by sugar density gradient centrifugation. In addition, by binding a liposome having an affinity to a target cell, the efficiency of gene transfer into the cell can be increased. As those having an affinity for a target cell, for example, an antibody,
And ligands such as receptors. The HST-1 DNA construct may take any form as long as it can be expressed in vivo. Preferred forms include in vivo stable forms such as plasmids.

When administered by the in vivo method, HST-1D
NA structures can usually be administered intravenously,
It can also be administered intraarterially, subcutaneously, intradermally, intramuscularly, topically, intraperitoneally, rectally, transdermally, transnasally, orally, and the like.
Intravenous preparations include, for example, injections such as solutions and suspensions, and can be produced by a usual method. That is, it can be produced by dissolving or suspending the HST-1 DNA structure in a pharmaceutically acceptable carrier or diluent. Pharmaceutically acceptable carriers or diluents include, for example, distilled water, physiological saline, dextrose aqueous solution and the like. If necessary, to these, as auxiliary agents, for example, pH adjusters, buffers (sodium acetate, sodium lactate, sodium citrate, etc.), stabilizers (human serum albumin, sorbitol, polyethylene glycol, amino acids, etc.), Dissolution aids (sorbitan monolaurate, triethanolamine oleate, etc.), tonicity regulators (sodium chloride, potassium chloride, calcium chloride, glucose, etc.), preservatives (benzalkonium chloride, benzyl alcohol, phenol, etc.), Sedative
(Such as procaine hydrochloride) and the like. The solution or suspension thus produced can be packaged as it is, or can be lyophilized and dissolved in sterile physiological saline before administration for use. This preparation can be made into a sustained-release preparation using a biocompatible material such as collagen, and can be prepared by a usual method (WO 97/02
047). The sustained-release preparation can be administered by various methods depending on the site of the disorder to be treated and its condition. For example, it can be administered subcutaneously or intramuscularly. In the case of a disorder such as a brain, the drug can be directly administered to a target site.

The dose and frequency of administration of the HST-1 DNA construct in the in vivo method generally vary depending on the animal species to be treated, the administration route, the degree of symptoms, age, body weight, sex, and the like.
The HST-1 DNA construct is usually used in an adult (body weight 60 kg) in a range of about 0.0001 mg to about 100 mg, preferably about 0.0005 m.
g to about 50 mg, more preferably about 0.001 mg to about 10 mg, administered once or more times a day or by continuous infusion. Also, administration once every two days to two months is possible.

Examples of the method for introducing the HST-1 DNA construct into cells in the ex vivo method include a microinjection method, a calcium phosphate method, and an electroporation method. In general, the introduction of this gene is preferably performed using a retroviral vector (Hum
an Gene Therapy, 2 , 251-256 (1991), Science, 243 , 2
20-222 (1989)). In general, in vivo methods are preferred because they are less expensive, easier to handle, and easier to control the expression period than ex vivo methods. The administered HST-1 DNA construct produces the HST-1 protein or its equivalent in vivo and shows its therapeutic and / or prophylactic effect. In general,
The HST-1 DNA construct exhibits the above-mentioned efficacy for a longer period of time than the HST-1 protein or its equivalent.

The HST-1 protein and an equivalent of the HST-1 protein can be usually administered intravenously, but may be administered intraarterially, subcutaneously, intradermally, intramuscularly, topically, intraperitoneally, orally,
It can also be administered rectally, transdermally, transnasally, and the like. Intravenous preparations include, for example, injections such as solutions and suspensions, and can be produced by a usual method.
That is, it can be produced by dissolving or suspending the HST-1 protein and an equivalent of the HST-1 protein in a pharmaceutically acceptable carrier or diluent. Pharmaceutically acceptable carriers or diluents include, for example,
Examples include distilled water, physiological saline, dextrose aqueous solution and the like. If necessary, to these, as auxiliary agents, for example, pH adjusters, buffers (sodium acetate, sodium lactate, sodium citrate, etc.), stabilizers (human serum albumin, sorbitol, polyethylene glycol, amino acids, etc.), Dissolution aids (sorbitan monolaurate, triethanolamine oleate, etc.), tonicity regulators (sodium chloride, potassium chloride, calcium chloride, glucose, etc.), preservatives (benzalkonium chloride, benzyl alcohol, phenol, etc.), Sedatives (eg, procaine hydrochloride) can be added. The solution or suspension thus produced can be packaged as it is, or can be lyophilized and dissolved in sterile physiological saline before administration for use.

Examples of preparations for subcutaneous, intramuscular, topical, intraperitoneal, intrarectal, transdermal and nasal administration include, for example, injections such as solutions, suspensions and emulsions, transrectal suppositories, transdermal Agents (oils, creams, lotions and the like) and the like. These preparations can be made into sustained-release preparations using a biocompatible material such as collagen, and can be produced by a usual method. Examples of oral administration preparations include powders, tablets, pills, capsules, granules, fine granules,
Suspensions, emulsions, syrups, solutions and the like can be mentioned. These preparations can be manufactured according to a usual method using a usual carrier or diluent. Dosage and frequency of HST-1 protein and HST-1 protein equivalents generally depend on the animal species to be treated, the route of administration, the severity of the symptoms, the age, weight, sex, and the like. HST-1
Protein and HST-1 protein equivalents are usually
For an adult (body weight 60 kg), about 0.0001 mg to about 200 mg
mg, preferably about 0.0005 mg to about 100 mg, more preferably about 0.002 mg to about 50 mg, administered once or more times a day or by continuous infusion. Also,
Administration once every two days to one month is also possible.

[0020]

The present invention will be described below in detail with reference to examples.
However, the present invention is not limited by these Examples.
Not. Example 1Introduction of HST-1 gene into adenovirus vector  As adenovirus vectors for recombination, E1A, E1
Adenovirus type 5 lacking the B and E3 regions (ATCC
No. VR-5) was used. This cassette cosmid vec
Promoter (pAdex1w) contains the SRα promoter
-, Human HST-1 cDNA (Proc. Natl. Acad. Sci. USA, 84, 2
980-2984 (1987)) and the polyA signal sequence of SV40.
Was. A recombinant vector containing HST-1 (Adex1HST-1L) is
o Recombination method (J. Virol.,54, 711-719 (1985); Proc. N
atl. Acad. Sci. USA,93, 1320 (1996); Cell,13, 18
1-188 (1978)). Collection of this viral vector
For gain, propagate this viral vector in 293 cells
And cultured until 293 cells became confluent
Thereafter, the cells were frozen and thawed five times. By centrifugation,
Remove cell debris and remove supernatant containing virus particles to -8
Stored at 0 ° C. To further increase virus concentration,
Centrifugation using a cesium chloride gradient (Jpn. Med. Sci.
 Biol.,7, 157-166 (1994)). Desalination
After, 1 × 10^Ten pfu (plaque forming unit) /
Stored at −80 ° C. to a virus concentration of mL.

Embodiment 2Evaluation of prevention effect of HST-1 by survival rate of mice exposed to radiation
Price test  Groups of 15 8-week-old C57BL / 6 mice (female)
And divided into three groups. Adenovirus vector described in Example 1
Tar (10⁹pfu / mouse) 3 days before irradiation
Administered intraperitoneally. Wild-type adenovirus as control
The vector was administered simultaneously. The test group and control group
9 gray (900 rad) radiation, 100% lethal dose to the us
Was irradiated. 20 days after irradiation for cell fraction in peripheral blood
Observed later. The survival rate was observed for 30 days after irradiation. Bone marrow, spleen
Tissue changes in the gut, intestine, etc., and intestinal bleeding
Observed after a day.

(1) Cell fractionation in peripheral blood

[Table 1] As shown in Table 1, the control group had almost no peripheral blood cells after irradiation, whereas the test group had recovered to the same level as the peripheral blood cell number of non-irradiated mice. In addition, remarkable recovery of the leukocyte count was observed as compared with the absence of cells in the control group.

(2) Survival rate The results of the survival rate are shown in FIG. The test group shows a remarkable life-prolonging effect, and also shows an excellent effect as compared with the control group.
That is, in the control group, 1 out of 15 animals 30 days after irradiation
Only the animals survived, but the test group survived 13 out of 15 animals 30 days after irradiation.

(3) Histological changes in bone marrow, spleen, and intestinal tract The results of histopathological changes are shown in FIGS. The test group
The tissue construction was close to normal, and almost no necrosis or shedding was observed. In contrast, widespread necrosis, shedding, and degeneration were remarkably observed in the control group, and severe degeneration occurred particularly in the bone marrow. In the spleen, atrophy was remarkable in the control group. In the control group, intestinal tract not only changes in histopathology,
Autopsy findings showed severe bleeding. On the other hand, no bleeding was observed in the test group. From these results, HS
The T-1 gene and HST-1 protein were shown to be effective in preventing tissue damage due to radiation.

Embodiment 3Evaluation of therapeutic effect of HST-1 gene on radiation-exposed mice
test  A group of 10 8-week-old C57BL / 6 mice (female)
And divided into three groups. Mice in both test and control groups
Illuminated with 9 gray (900 rad), a 100% lethal dose
Fired. Six hours after the irradiation, the test group was subjected to the method described in Example 1.
Denovirus vector (10⁹pfu / mouse) intraperitoneally.
Was. Wild-type adenovirus vector is simultaneously used for the control group
Was administered. For the cell fraction in peripheral blood,
Observed after 0 days. Viability was monitored for 30 days after irradiation.
Was.

(1) Cell fractionation in peripheral blood

[Table 2] As shown in Table 2, the control group had almost no peripheral blood cells after irradiation, whereas the test group had recovered about half of the peripheral blood cell count and platelet count of non-irradiated mice. Was. In particular, the recovery of the white blood cell count was remarkable.

(2) Survival rate The results of the survival rate are shown in FIG. In the control group, all 10 animals died by 23 days after irradiation,
In the test group, 2 out of 10 animals survived 30 days after irradiation, and a significant difference (p = 0.04) was observed in the survival effect. These results revealed that the HST-1 gene, HST-1 protein and the like are effective for treating tissue damage caused by radiation.

REFERENCE EXAMPLE 1 Method for Producing Sustained-Release HST-1 Gene Preparation (Gel Form) 100 μg / ml pCAHST-1 solution (200 μl) and 10 mg / ml glucose in 0.1 w / w% atelocollagen solution (500 mg) Then, a sucrose or monosodium glutamate solution (500 μl) was added, and after mixing, the mixture was kept at 37 ° C. to prepare a gel-like gene preparation. Reference Example 2 Method for producing sustained release HST-1 gene preparation (rod-like) 0.86 w / w% atelocollagen solution (29.1 g)
(60 g), a 100 μg / ml pCAHST-1 solution (80 ml) and an 11 mg / ml glucose solution (10 ml) were added to the mixture and mixed. After freeze-drying the obtained solution, an appropriate amount of distilled water was added to the freeze-dried product and kneaded. Thereafter, the kneaded product was put into a syringe, extruded, and further dried to obtain a pCAHST-1 yield of 74%.
Was obtained. That is, 17 μg of pCA per mg
A bar-shaped gene preparation containing HST-1 and 300 μg of glucose was prepared.

[0029]

According to the present invention, the HST-1 DNA construct, HST-1
An agent for promoting the growth and / or differentiation of stem cells, germ cells, hair cells, intestinal cells, villous tissues, mucous membranes, glomerular endothelial cells and spleen cells containing the protein or its equivalent can be provided. In addition, prevention of tissue damage (excluding thrombocytopenia) due to anticancer therapy or radiation containing the HST-1 DNA construct, HST-1 protein or its equivalent, and / or
Alternatively, a therapeutic agent can be provided. According to the present invention, not only the side effects during cancer treatment are reduced, but also the radiation treatment itself can be more effectively performed.

[Brief description of the drawings]

FIG. 1 shows the survival rate of mice exposed to radiation in the preventive effect evaluation test of Example 2 (2).

FIG. 2 is a micrograph (about × 40) of a bone marrow tissue specimen of a radiation-exposed mouse of a control group in the preventive effect evaluation test of Example 2 (3).

FIG. 3 is a micrograph (about × 40) of a bone marrow tissue sample of a radiation-exposed mouse in the administration group in the preventive effect evaluation test of Example 2 (3).

FIG. 4 is a photomicrograph (about × 200) of a bone marrow tissue specimen of a radiation-exposed mouse of a control group in the preventive effect evaluation test of Example 2 (3).

FIG. 5 is a micrograph (about × 200) of a bone marrow tissue specimen of a radiation-exposed mouse in the administration group in the preventive effect evaluation test of Example 2 (3).

FIG. 6 is a micrograph (approximately × 40) of a spleen tissue specimen of a control group in a radiation-exposed mouse in the preventive effect evaluation test of Example 2 (3).

FIG. 7 is a micrograph (about × 40) of a spleen tissue specimen of a radiation-exposed mouse in the administration group in the preventive effect evaluation test of Example 2 (3).

FIG. 8 is a micrograph (about × 40) of an intestinal tissue specimen of a radiation-exposed mouse of a control group in the preventive effect evaluation test of Example 2 (3).

FIG. 9 is a photomicrograph (about × 40) of an intestinal tissue sample of a radiation-exposed mouse in the administration group in the preventive effect evaluation test of Example 2 (3).

FIG. 10 shows the survival rate of radiation-exposed mice in the therapeutic effect evaluation test of Example 3 (2).

Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat II (reference) C07K 14/50 C07K 14/50 C12N 15/09 C12N 15/00 A (72) Inventor Hiromi Sakamoto 5 Tsukiji, Chuo-ku, Tokyo −5-1 Inside the National Cancer Center Research Institute (72) Inventor Yasushi Takahama 355, Nosagawa-gawa Village, Yoshino-gun, Nara Pref.

Claims (11)

[Claims] Claims: 1. An HST-1 DNA construct, a stem cell, a germ cell, a hair mother cell containing an HST-1 protein or an equivalent thereof,
An agent for promoting the growth and / or differentiation of intestinal cells, villous tissue, mucous membrane, glomerular endothelial cells and spleen cells. 2. A preventive and / or therapeutic agent for tissue damage (excluding thrombocytopenia) due to anticancer therapy or radiation, comprising an HST-1 DNA construct, an HST-1 protein or an equivalent thereof. 3. The method of claim 2, wherein the anticancer therapy is radiation therapy.
The prophylactic and / or therapeutic agent as described above. 4. The preventive and / or therapeutic agent according to claim 2, wherein the damaged tissue is a tissue composed of hematopoietic stem cells, intestinal stem cells, or neural stem cells. 5. The preventive and / or therapeutic agent according to claim 2, wherein the damaged tissue is bone marrow or intestine. 6. The preventive and / or therapeutic agent according to claim 2, wherein the tissue disorder is leukopenia, anemia, erythropenia or mucositis. 7. The preventive and / or therapeutic agent according to claim 2, wherein the tissue damage is bone marrow necrosis or a decrease in peripheral blood cells. 8. The preventive and / or therapeutic agent according to claim 2, wherein the tissue disorder is intestinal bleeding, reduction of villous tissue, or a secondary infection disorder. 9. The method according to claim 1, which comprises an HST-1 DNA construct.
8. The medicament according to any one of 8 above. 10. The medicament according to claim 9, wherein the HST-1 DNA construct is introduced into a vector. 11. The medicament according to claim 9, wherein the HST-1 DNA construct is introduced into an adenovirus vector or a plasmid vector.