WO1994004175A1

WO1994004175A1 - Hgf production promoter

Info

Publication number: WO1994004175A1
Application number: PCT/JP1993/001177
Authority: WO
Inventors: Toshikazu Nakamura
Original assignee: Toshikazu Nakamura
Priority date: 1992-08-24
Filing date: 1993-08-23
Publication date: 1994-03-03

Abstract

A promoter for the production of HGF (hepatocyte growth factor) having the activity of promoting the cure of injuries of tissues and organs of living organisms. The promoter contains as the active ingredient at least one member selected from the group consisting of interleukin-1α, interleukin-1β, heparin and derivatives thereof (such as low-molecular-weight heparin), heparan sulfate, prostaglandin E1, prostaglandin E2, prostaglandin I2, derivatives of the prostaglandins, and clathrate compounds of the prostaglandins and derivatives thereof. This promoter is useful for curing wounds and so forth, because it has the activity of promoting the production of HGF in HGF-producing cells.

Description

Description H GF production promoter Technical field

The present invention relates to an agent for promoting HGF (Hepatoc to Growth Factor, hepatocyte growth factor) production, and more particularly to a substance that promotes HGF production of HGF-producing cells. Conventional technology

HGF is a protein that the present inventors have found in regenerated liver rat serum as a factor for growing mature hepatocytes in vitro (Biochem Biophys Res Commun, 122, 1450, 1984). The present inventors have further succeeded in isolating HGF from rat platelets (Proc. Natl. Acad. Sci, 83, 6489, 1986, FFBS Letter, 22, 311, 1987), and identified the amino acid sequence of the same. Department decided. Furthermore, the present inventors performed human and rat-derived HGF cDNA cloning based on the elucidated HGF amino acid sequence, and recombined the cDNA into animal tissues to proliferate hepatocyte cells. The factor was successfully obtained as a protein (human HGF: Nature, 342, 440, 1989; rat HGF: Proc. Natl. Acad. Sci, 87, 3200, 1990).

The molecular weight of HGF is 82-85 kD by SDS-polyacrylamide gel electrophoresis. Rat HGF molecules have a heterodimer structure in which an α chain consisting of 463 amino acid residues and a 9 chain consisting of 233 amino acid residues are bridged by a single disulfide bond. Each chain has two darcosamine-type sugar chain binding sites. Human HGF also has almost the same physiological activity. It consists of an α-chain consisting of 463 amino acid residues and a / chain consisting of 234 amino acid residues. The α chain has four kringle structures similar to the fibrinolytic enzyme plasmin, and the amino acid sequence of the ^ chain is approximately 37% homologous to the 一 chain of plasmin, which has serine mouth opening activity. Having. The homology of the amino acid sequences of rat HGF and human HGF was 91.6%, Has a very high homology of 88.9%, and their activities are completely compatible.

HGF, which was discovered as a factor that specifically proliferates hepatocytes, has been shown to show various activities in vivo based on recent research results by the present inventors and other researchers. Expectations are growing for its application not only as a drug but also as a therapeutic agent for humans and animals.

The present inventors have clarified that HGF acts not only on hepatocytes but also on epithelial cells as a growth factor, and have achieved several inventions. In Japanese Patent Application Laid-Open No. 4-49246, HGF promotes the proliferation of renal proximal tubule cells, and thus aims to develop its application as a therapeutic agent for renal diseases. In No. 158, HGF promotes the growth of normal epithelial cells such as melanocytes and keratinocytes, and is being developed as an epithelial cell promoting agent for the treatment of wounds, skin ulcers, and hair root cell proliferating agents. Fulfilled and disclosed the details. In particular, HGF is more suitable for practical use because it does not have the canceration effect and cancer cell proliferation activity found in many other growth factors such as EGF. Furthermore, the present inventors have disclosed in Japanese Patent Application No. 3-140812, cancer cells such as Hep G2 cell line derived from human liver cancer of HGF and IM9 cell line derived from lymphoblastoid cancer. Utilizing its growth inhibitory activity, it disclosed that it can be used as an anticancer agent.

Also, in Japanese Patent Application No. 3-321241, HGF is useful as a side effect inhibitor for cancer therapy, which can reduce the damage to normal cells and tissues in cancer treatment and reduce or prevent side effects. It is disclosed that there is.

In addition, HGF is useful for the prevention and treatment of chronic and acute lung diseases such as pneumonia, emphysema, chronic obstructive pulmonary disease, pneumoconiosis, and swallowing pneumonia in Japanese Patent Application No. 4-298053. Is disclosed.

In addition, the present inventors have pointed out that HGF may be an entity of a neural inducing factor that contributes to the developmental stage of the central nervous system, and that it is useful for central diseases and useful for eye diseases, that is, Its usefulness in tongue has been clarified. These are mentioned in various documents of the present inventors, and their usefulness is clear (Medical immunology, 26, No 1, 1993, etc.) _; In some of the recent reports of the present inventors, Biochemical and Research Co., Ltd. uin. 191, No. 2, 1993 clarifies that the usefulness of HGF extends to the digestive tract.

In addition, since HGF is involved in the formation of osteoclasts, it inhibits osteoclast resorption, promotes osteoblast formation, and is useful for bone diseases, and it promotes osteoarthritis through the promotion of proteogliin synthesis. The usefulness of HGF as a drug has been clarified, and the potential of HGF as a drug is not only liver disease, but also kidney disease, skin disease, blood disease, eye disease, lung disease, stomach, duodenal disease, It is becoming clear that the disease is diverse, such as cancer and related diseases and bone diseases.

An even more important point in considering the usefulness of HGF as a drug is that HGF promotes growth only in cells that have entered the G1 phase, the proliferative phase, and does not act on cells in the GO phase, that is, the quiescent phase. It is. This means that the growth and regeneration of damaged tissue is promoted, but has no effect on undamaged tissue. Therefore, it is considered that even if HGF is excessively administered or reaches non-affected areas via blood, etc., it does not induce canceration in normal tissues or cause excessive proliferation. .

As described above, since HGF promotes proliferation of epithelial cells as well as hepatocytes widely and has cancer cell growth inhibitory activity, it is expected that HGF acts on tissue injury healing in vivo. . HGF-producing cells are not epithelial cells themselves, but mainly mesenchymal cells such as K upffer cells and sinusoidal vascular endothelial cells in the liver, capillary endothelial cells in the kidney, and alveolar macrophage vascular endothelial cells in the lung. It has been elucidated that it is produced by cells, and it has been clarified that the so-called paracrine mechanism, in which HGF is supplied from neighboring cells as needed, has been established.

However, when the liver and kidneys are injured, HGF production is also increased in non-injured organs, such as the lungs, so it may be that HGF is supplied by the so-called endocrine mechanism: That is, substances that promote HGF production are secreted from the injured tissue. It is thought to reach the HGF-producing cells via blood and release HGF stored in the cells, or to initiate new production.

Of course, HGF is administered for the purpose of wound healing and renal regeneration, but if a substance that promotes HGF production is administered, the same effect is expected to be obtained with a smaller dose and the number of administrations. You. More specifically, compared to the case where HGF is directly administered as a treatment for tissue injury, administration of the HGF production promoting substance can maintain the blood concentration of HGF for a longer period of time. It is expected that the dose and frequency of administration can be reduced. Although it is difficult to absorb HGF transdermally, if the substance that promotes HGF production has transdermal absorbability, it becomes possible to obtain a preparation that promotes HGF production by transdermal absorption. As formulation and use become easier, application can be expected to expand.

On the other hand, living organisms are constantly injured by microbes, whether external or internal, in all tissues and organs, including the respiratory tract, skin, and digestive organs. In contrast, recruiting all functions, including cell growth factors such as HGF, FGF, EGF, and PDGF, and repairing functional tissue cells of organs and matrix cells is one important homeostasis. As a factor, it has been thought that there is a central factor in the tissue that activates and regulates the compensation network.

The HGF production promoting factor is useful not only for studying the function of compensating for organ damage such as liver and kidney and for studying the mechanism of HGF expression, but also for healing tissue and organ damage of living organisms having HGF receptor. However, it is clear that it is very useful as a therapeutic drug. Its usefulness is evident in view of the usefulness of HGF described above.For example, in the case of the liver, administration of HGF can regenerate only hepatic parenchymal cells during hepatectomy or organ damage due to hepatitis In contrast, by administering HGF production promoting factors that control the entire repair function of organ injury, not only parenchymal cells but also non-parenchymal cells, such as supporting tissues such as peri-sinusoidal connective tissue, can be regenerated throughout Will be accelerated and true restoration will be promoted. In other words, if the HGF production promoting factor can be used as a therapeutic agent, the injury will be treated much more mildly and promptly than the conventional use of various cell growth factors alone. And its usefulness is immense.

From this point of view, the present inventors have conducted intensive studies on substances that promote HGF production and found that interleukin-1α, interleukin-1 ^ 9, heparin and derivatives thereof, heparan sulfate, prostaglandin Ε, and a derivative conductor book found that there is a prostaglandin E ₂ and derivatives thereof, prostaglandin 1 ₂ and their derivatives, as well as significant HGF production promoting action in clathrate thereof prostaglandins or derivatives thereof Completed the invention. That is, an object of the present invention is to provide a therapeutic agent for various diseases as described above by having an HGF production promoting action. Disclosure of the invention

The present invention relates to interleukin-1α, interleukin-1 / 9, heparin or a derivative thereof, heparan sulfate, prostaglandin _II , prostaglandin II ₂ , prostaglandin I2, and these prostaglandins. And a prostaglandin or an inclusion compound of these derivatives as an active ingredient. Two or more of these active ingredients may be used in combination. Another invention is a method for treating liver disease or the like, which comprises administering an effective amount of the above compound to a human. BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 is a diagram showing the relationship between the concentration of interleukin-1α and interleukin-11 ^ and the amount of HGF production in Example 1. In the figure, Hata indicates interleukin-1α and 、 indicates Interleukin-1 1 S is shown.

FIG. 2 is a graph showing the relationship between heparin concentration and HGF production in Example 2. FIG. 3 is a graph showing the relationship between prostaglandin (hereinafter referred to as PG) concentration and HGF production amount in Example 3. In the figure,拿is P GE 〇 is P GE _2, mouth 〇 P 2 5 07, △ denotes a ON04 1 4 8 3. FIG. 4 is a diagram showing the relationship between the culture time and the amount of HGF production in Example 5. In the figure, Qin the presence of P GE _2, 〇 indicates a control.

FIG. 5 is a graph showing a time-dependent change in LI (labeling index) when lmgZkg of heparin was administered to a rat in Example 6. In the figure, · indicates a system to which heparin was administered, and 〇 indicates a control.

FIG. 6 is a graph showing the change over time in blood HGF concentration when 1 mg / kg of heparin was administered to a rat in Example 6. In the figure, 拿 indicates the system to which heparin was administered, and 〇 indicates the control. BEST MODE FOR CARRYING OUT THE INVENTION

Interleukin-1a and interleukin-11 (9 are known substances, which are the active ingredients of the HGF production promoter of the present invention, are isolated from mammalian tissues and the like according to a conventional method. Any of the above-mentioned substances may be used, such as those prepared by genetic engineering means, etc. Further, the above-mentioned substances may be substances in which a part of the amino acid sequence has been deleted or substituted with other amino acids, or other amino acid sequences. , A substance in which one or more amino acids are bound to the N-terminus and Z- or C-terminus, a substance in which a sugar chain is added, substituted or deleted, etc.

Heparin and heparan sulfate are also known substances, and those isolated from mammalian tissues or the like according to a conventional method can be used. Heparin derivatives include low-molecular-weight heparin (average molecular weight of about 440,600) obtained by chemical treatment of heparin, and functional groups of heparin (for example, hydroxyl group). , Carboxyl group, etc.) and a chemical treatment (eg, acylation, alkylation, esterification, amidation, etc.) into which a substituent is introduced. Pas phosphorus derivatives heparin and to the of the above, a salt thereof (e.g., alkali metal salts) may be in the form of _£ Further, the active ingredient of the present invention, PGE,? 〇 5 ₂ and 〇 1 ₂ is the substance of the publicly known, for example, can be used commercially. The derivatives of these PGs include compounds that have been subjected to various chemical modifications in order to improve stability, enhance action, etc., and are PGs and derivatives thereof having a carboxyl group. In the case of a compound, it may be in the form of a pharmaceutically acceptable salt. Examples of inclusion compounds of the above PGs and derivatives thereof include inclusion compounds of these compounds with α-cyclodextrin, ^ -cyclodextrin and the like.

Examples of the above PG derivatives and clathrate compounds include, for example,

3—3 6 4 5 8 5 4—3 2 7 7 3 5 7—2 0 3 0 5; Japanese Unexamined Patent Publication No. 49-47 7 3 5 2 5 1—1 0 1 9 6 1 5 1—1 2 2 04 0 5 2-2 7 7 5 3 5 2-4 2 8 5 6 5 3—8 4 9 4 2 5 4—4 4 6 3 9 5 5—1 0 0 3 6 0 5 8—4 7 6 25 58-20 39 64, 58-20 39 91 1 59-20 27 67, etc., or the methods described in these publications or compounds prepared according to them Is mentioned. Preferred examples of PG derivatives and clathrates include, for example,

4 1 4 8 3 and the like.

The accelerator of the present invention can take various formulation forms (for example, liquids, solids, capsules, etc.), and generally, the active ingredients interleukin-1α, interleukin-1β, heparin or heparin Only one or two or more components selected from the group consisting of derivatives thereof, heparan sulfate, PGE PGE ₂ PGI ₂ , derivatives of these PGs, and clathrates of these PGs or derivatives thereof, or They are used as injections with conventional carriers, or as external drugs with conventional carriers. The injection can be prepared by a conventional method. For example, the above active ingredient is dissolved in an appropriate solvent (eg, sterile water, buffer solution, physiological saline, etc.), and then filtered through a filter or the like. And then filled in a sterile container. The content of the active ingredient in the injection is appropriately adjusted. For external preparations, for example, they are formulated into ointments, gels, liquids, etc., and the active ingredient content in the preparations should be adjusted as appropriate according to the disease, site of application, etc. But it can.

At the time of formulation, a stabilizer is preferably added, and examples of the stabilizer include albumin, globulin, gelatin, mannitol, glucose, dextran, ethylene glycol and the like. Further, it may contain additives necessary for preparation, for example, excipients, solubilizers, antioxidants, soothing agents, tonicity agents and the like. In the case of a liquid preparation, it is desirable to store it after freezing or freezing it by lyophilization. The freeze-dried preparation is used after reconstitution by adding distilled water for injection at the time of use.

The accelerator of the present invention can be administered by an appropriate administration route depending on the form of the preparation. For example, it can be administered in the form of injection, intravenously, arterial, subcutaneously, intramuscularly and the like. The dose is appropriately adjusted according to the patient's condition, age, weight, and the like.

HGF has the effect of promoting the healing of injuries to tissues and organs of living organisms. Therefore, the HGF production promoter of the present invention is, for example, a therapeutic agent for liver disease, a therapeutic agent for renal disease, or a therapeutic agent for wound as described above. It has applications such as skin ulcer treatment agent, hair root cell proliferative agent, anticancer agent, lung injury treatment agent, side effect preventive agent for cancer therapy, and more specifically, for example, liver disease (eg, hepatitis, cirrhosis, liver failure) , Liver regeneration after surgery, etc.), renal disease (eg, glomerulonephritis, renal failure, renal anemia, diabetic nephropathy, renal injury after drug administration, etc.), skin disease (eg, vitiligo disease, burn) , Floor rub, skin ulcer, baldness, etc., blood disease (eg, thrombocytopenia, bone marrow transplant, etc.), eye disease (eg, corneal ulcer, etc.), lung disease

(E.g., pneumonia, emphysema, pulmonary tuberculosis, chronic obstructive pulmonary disease, pneumoconiosis, pulmonary fibrosis, etc.), gastroduodenal diseases (e.g., gastritis, gastric ulcer, duodenal ulcer, etc.), cancer diseases and related diseases (e.g., various cancers, Side effects from cancer therapy, such as prevention of hepatotoxicity, nephrotoxicity, nausea, vomiting, thrombocytopenia, hair loss, etc., bone diseases (eg, osteoporosis, osteodysplasia, osteoarthritis, etc.), central diseases (eg, It is useful for prevention and treatment of diseases such as neuropathy. Industrial applicability

According to the promoter of the present invention, HGF production can be promoted, Can promote the healing of injuries of tissues and organs, such as liver disease, kidney disease, skin disease, blood disease, eye disease, lung disease, gastroduodenal disease, cancer And its related diseases, bone diseases, central diseases and the like. Example

Hereinafter, the present invention will be described in more detail with reference to Examples, but the present invention is not limited to these Examples.

Example 1

HGF production promoting activity of interleukin-1 and interleukin-1β

Human skin fibroblasts were seeded on a 24-well plate at a density of 5 × 10 ⁴ cells / cm ² and cultured for 24 hours. The medium was replaced with fresh DMEM (Dulbecco, modified Eagle's medium) supplemented with 1% fetal calf serum (FCS), and then interleukin-1α or interleukin-1β was added. Cultured for hours. The HGF produced by the culture was measured by an enzyme immunoassay using a perforated anti-human HGF polyclonal antibody according to a conventional method.

Figure 1 shows the results. In the figure, · indicates interleukin-1 1 and 〇 indicates interleukin-1 1 ^. As shown in FIG. 1, it was found that both interleukin-11α and interleukin-11β significantly promote HGF production even at low concentrations.

Example 2

Heparin HGF production promoting activity

MRC-5 human embryonic lung fibroblasts were seeded on a 24-well plate at a density of 5 × 10 ⁴ cells / cm ² and cultured for 24 hours. After the medium was replaced with fresh DMEM supplemented with 1% FCS, heparin dissolved in phosphate buffer was added, and the cells were cultured for 24 hours. HGF produced by the culture was measured by an enzyme immunoassay using a rabbit heron anti-human HGF polyclonal antibody according to a standard method. Figure 2 shows the results. As shown in FIG. 2, heparin was found to significantly promote HGF production even at low concentrations.

Example 3

HGF production promoting activity of PGs

Human skin fibroblasts were seeded on a 24-well plate at 5 × 10 ⁴ cells / cm ² and cultured for 24 hours. The medium was replaced with 1% FCS new Korea was added ^{^{DMEM, 1 0- 12 1 0- 6}} M PG such (i.e., is a P GE, derivatives or inclusion compound of P GE ₂ and PEI ₂ 0P2507 and ONO41483) were added and cultured for 24 hours. The HGF produced by the culture was measured by an enzyme immunoassay using an anti-human rabbit HGF polyclonal antibody according to a conventional method.

Figure 3 shows the results. In FIG. 3, PGE P indicates PGE ₂ port は Ρ 2507 Δ indicates ON041483 respectively.

As shown in FIG. _{3, P GE, P GE 2} OP 2 5 07 and 〇_NO 4 1 4 8 3 are all 1 0- ^S HGF production promoting action from M was observed, with 1 0- ⁶ M Showed maximum promotion. Promoting production at this time, 3 0.6 times P GE ₁ as compared to the control, 6.7-fold with PGE _2, 0 2 5 07 7 5.4 times, 0 N 04 1 4 8 3 2 It was 0.7 times.

Example 4

Effects of various PGs on HGF production in human dermal fibroblasts and MRC-5 cells

Human skin fibroblasts and MRC-5 cells were seeded at 5 × 10 ⁴ cells Z cm ² on 24-well plates and cultured for 24 hours. The medium was replaced with 1% FC S Fresh was added pressure of DMEM, PG such 1 0- ⁶ M a _{(PGE, PGE 2 0 2 5} 07 and Rei_1 \ ⁷ 04 1 4 8 3) was added The cells were cultured for 24 hours. The HGF produced by the culture was measured by an enzyme immunoassay using a perforated anti-human HGF polyclonal antibody according to a conventional method. The results are shown in Table 1.

As shown in Table 1, in human skin fibroblasts, it was exhibited significant HGF production promoting effect, PGE, and a derivative of PGI _z 〇 P 2 5 ◦ 7.

On the other hand, in MRC-5 cells, it was found that PGE and PGE ₂ had a slightly stronger HGF production promoting effect than PGI ₂ derivatives.

table 1

PGs (10 M) HGF production (ng gmg protein)

Human skin fibroblasts MRC-5 cells Control 0.0 2 5 3. δ

P G E, 3. 4 3 3 7 1. 8

PGE ₂ 0.7 5 3 8 5.2

0 P 2 5 0 7 8.6 3 5 6.7

0 N 04 1 4 8 3 2.3 3 1 8 3.3 Example 5

Time course of HGF production by PGΕ in human dermal fibroblasts

Human skin fibroblasts were seeded on a 24-well plate at 5 × 10 ⁴ cells / cm ² and cultured for 24 hours. The medium, after replaced with fresh DME M supplemented with 1% FCS, the presence or absence of PGE ₂ in 1 0- ⁶ M (Control port Lumpur), 3, 6, 9, 1 2 and Cultured for 24 hours. At each culturing time, HGF produced in the culture system was measured by an enzyme immunoassay using a rabbit heron anti-human HGF polyclonal antibody according to a conventional method. Figure 4 shows the results. In the figure, - is the presence of PGE _2, 〇 is shows the control.

As shown in Figure 4, compared to the control, 〇 £ ₂ ^ 〇? The production promoting effect was observed immediately after the addition, and the production of HGF increased. Furthermore, HGF production began to increase rapidly from 6 hours after addition, and reached a plateau in 12 to 24 hours.

Example 6

Effect of heparin administration on 30% partial hepatectomy rat

The effects of the administration of heparin to 30% partial hepatectomy rats (DXA synthesis change, blood HGF concentration change) were tested by the following methods. (1) Method

① Animal treatment

Using a male Wistar rat (weight 120-210 g), the abdomen was opened under ether anesthesia, and only the left lobe was inverted, ligated and excised. Almost 30% of the liver can be removed. Heparin was administered intraperitoneally immediately after hepatectomy and every 12 hours after resection, and sacrificed at 12, 24, 36, 48 and 72 hours. One hour before sacrifice, 5-promo 2'-deoxyperidine (BrdU) 5O mgZkg was intraperitoneally administered. At the time of sacrifice, after whole blood was collected from the inferior vena cava, the remaining liver was excised, weighed, a portion was fixed in 70% ethanol for tissue staining test, and the others were frozen and stored at 180 ° C. In addition, a system to which physiological saline was administered instead of heparin was used as a control.

② DNA synthesis

A part of the liver fixed with 70% ethanol was embedded in paraffin, and immunohistochemically stained with an anti-BrdU monoclonal antibody according to a conventional method. After staining, a photograph was taken in a 100-fold visual field, the ratio (%) of the number of stained cells in four randomly selected visual fields was determined, and the average was defined as a labeling index (hereinafter referred to as LI).

③ Blood HG F concentration

Plasma was prepared from whole blood collected at the time of sacrifice and stored at 4 ° C. The plasma was partially purified on a heparin-Sepharose column according to the procedure described below, and the HGF concentration was measured by an enzyme immunoassay using a polyclonal alfa α-rat HGF antibody.

(a) Dilute 1 ml of plasma with 4 ml of a low salt HS (Hartman's solution) buffer.

(b) _{Pass the} above sample through _{heparin-cephalo-scalar CL-6B (bed} volume 1 m〗) equilibrated with low salt concentration HS buffer.

(c) Wash the column with a low salt concentration HS buffer 5m].

(High salt concentration HS buffer (containing 2 M sodium chloride) δ rn] Recover the first 2 ml ₌

(e) Dialyze the collected eluate against a sufficient amount of WE solution overnight, and dialyze Then, the HGF concentration is measured by enzyme immunoassay.

(2) Result

The results tested by the above method were as follows.

① DNA synthesis

The time course of LI when heparin was administered at 1 mg / kg is shown in FIG. 5. As shown in FIG. 5, very strong promotion of DNA synthesis was observed 24 hours later. Also, slightly higher promotion of DNA synthesis than in the control was observed over 36 to 48 hours.

Next, Table 2 shows the relationship between the heparin dose and LI 24 hours later.

Table 2

As shown in Table 2, the LI value after 24 hours correlated with the heparin dose.

As shown in the above results, it was revealed that administration of heparin promoted DNA synthesis.

② Blood H G F concentration

Figure 6 shows the time course of blood HGF concentration when heparin was administered at 1 mg / kg. As shown in FIG. 6, the control showed a peak after 12 hours, and then gradually decreased. In contrast, when heparin was administered, a significant increase in blood HGF concentration was similarly observed after 12 hours, and an increase in blood HGF concentration was also observed after 48 hours. The increase in blood HGF concentration is observed after administration of _c . The two peaks are presumed to be due to the increase in blood HGF concentration due to different mechanisms of action (t) Example 7

Effect of low molecular weight heparin on 30% partial hepatectomy rat

A test similar to that of Example 6 was performed using low-molecular-weight heparin [Fragmin IV (trade name), average relative molecular weight of about 5,000] instead of heparin. Table 3 shows the LI values 24 hours and 36 hours after administration of lmgZkg of palin to the low molecule.

Table 3

As shown in Table 3, it was revealed that administration of small molecule heparin promoted DNA synthesis.

Example 8

Effect of administration of PGs on 30% partial hepatectomy rat

A test similar to that of Example 6 was performed using PGs (〇P—2507, PGE,) instead of heparin. The dosing method was subcutaneous administration to the back, PGs were administered immediately after hepatectomy and 12 hours later, and sacrificed 24 hours later.

Table 4 shows the LI value and the blood HGF concentration after 24 hours when PGs were administered at 100 g / kg or SOgZkg.

'' Table 4

As shown in Table 4, it was clarified that administration of PGs promoted DNA synthesis and increased blood HGF concentration.

Claims

The scope of the claims

1. interlink one interleukin one la, interleukin one 1/9, heparin or heparan sulfate, prostaglandins E ,, prostaglandin E ₂ derivatives to, to, prostaglandin 1 _2, induction of prostaglandins HGF containing as an active ingredient at least one selected from the group consisting of a conductor and an inclusion compound of these prostaglandins or derivatives thereof

(Hepatocyto Growth Factor) A production promoter.

2. Claims 1 used for the treatment of liver disease, kidney disease, skin disease, blood disease, eye disease, lung disease, gastroduodenal disease, cancer disease, cancer-related disease, bone disease or central disease The agent for promoting HGF production according to the above item.

3. The HGF production promoter according to claim 1, wherein the active ingredient is interleukin-11 or interleukin-11β.

4. The HGF production promoter according to claim 1, wherein the active ingredient is heparin or a derivative thereof or heparan sulfate.

5. active ingredient, prostaglandin E ,, prostaglandin E _2, prostaglandin 1 _2, derivatives thereof prostaglandins, and Ri by the group consisting of inclusion compounds of these prostaglandins or derivatives thereof 2. The HGF production promoter according to claim 1, which is at least one selected from the group.

6. The HGF production promoter according to claim 4, wherein the active ingredient is high molecular heparin or low molecular heparin.

7. interlink one interleukin one 1 shed, inter one interleukin one 1/3, heparin or heparan sulfate derivatives to, prostaglandin E have prostaglandin E _2, prostaglandin 1 _2, these prostaglandins A liver disease, a kidney disease, or a skin disease comprising administering to a human an effective amount of at least one compound selected from the group consisting of a derivative and an inclusion compound of these prostaglandins or derivatives thereof. A method for treating a blood disease, an eye disease, a lung disease, a gastroduodenal disease, a cancer disease, a cancer-related disease, a bone disease or a central disease _: