JP3159441B2 - Prevention and treatment of cytopenias - Google Patents

Description

Description: TECHNICAL FIELD The present invention relates to a novel medicine for humans or animals, and more specifically, to the side effects caused by the administration of various chemotherapeutic agents including anticancer agents. Agents that have protective and therapeutic effects, especially those that promote the recovery of hematopoietic dysfunction caused by the administration of chemotherapeutic agents such as anticancer agents, and prevent or treat disorders of a decrease in blood cell components such as leukocytes, reticulocytes, and platelets. About. Furthermore, the present invention relates to an agent for preventing and treating disorders of reduction of blood cell components such as leukocytes, reticulocytes, and platelets due to serious diseases such as cancer and viral diseases such as human immunodeficiency virus.

(Background technology)

In pharmacotherapy of various diseases, side effects such as bone marrow suppression and hematopoietic dysfunction are taken seriously. Leukopenia, leukocyte disorders such as granulocytopenia, anemia, erythrocyte disorders such as erythropenia or thrombocytopenia, etc., various drugs, especially anticancer agents such as anticancer agents, immunosuppressants, antibiotics, antivirus It is often caused by the administration of chemotherapeutic agents that cause cancer, etc., and from those disorders or symptoms, humans,
It has been desired to develop drugs for restoring animals and for protecting and treating such side effects. However, there are few such drugs, and some of them are clinically applied, such as cepharanthin and inosine, but their effects are not necessarily satisfactory.

Development of prophylactic / therapeutic agents capable of protecting various hematological disorders caused by administration of chemotherapeutic agents and the like requires the administration of chemotherapeutic agents and the like that can be used for humans or animals in drug therapy such as chemotherapy. This is extremely important because it allows the dose to be increased and further enhances the therapeutic effect of chemotherapy and drug therapy for cancer, infectious diseases and various diseases.

[Disclosure of the Invention]

The present inventors, for the purpose of developing a drug that enhances the body defense ability originally present in the living body, for various highly safe natural peptides, leukocytes, reticulocytes, blood cell components such as platelets As a result of an extensive search for substances useful as prophylactic / therapeutic agents to protect against impaired blood pressure and hematopoietic dysfunction, serum thymic factor (Facteur thymique ser
ique, hereinafter abbreviated as FTS) nonapeptide and its derivatives or salts thereof clearly protect and prevent the reduction of blood cell components such as leukocytes, reticulocytes and platelets induced in mice administered with anticancer agents. , Also found to be treated. The present invention has been made based on such findings, and provides a prophylactic / therapeutic agent for protecting various blood disorders caused by in vivo administration of chemotherapeutic agents such as anticancer agents, antiviral agents, and antibacterial agents. It is.

Further, the present invention itself provides a preventive / therapeutic agent for a cancer accompanied by a decrease in blood cell components such as leukocytes, reticulocytes, and platelets, and a viral disease such as a human immunodeficiency virus.

The present inventors previously found nonapeptide known as FTS multiple sclerosis, Guillain-Barre syndrome, inflammatory neuritis,
It has been found that it is suitable as a therapeutic agent for various diseases associated with immunodeficiency such as polyneuritis and other immune demyelinating diseases, and such a therapeutic agent has been provided (JP-A-58-5222).
5), the fact that nonapeptide known as FTS has a prophylactic and therapeutic effect to protect against impairment of blood cell components such as leukocytes, reticulocytes, and platelets caused by chemotherapeutic agents that cancer cancer drugs. It is completely unexpected from the prior art and has been found for the first time by the present inventors. Thus, the present invention provides a nonapeptide having the following amino acid sequence: Or an ester at the carboxyl group of asparagine at the C-terminus, an amide, or a leukocyte characterized by containing a pharmaceutically acceptable salt of each of these compounds or a combination of each of these compounds with a metal as an active ingredient, It is intended to provide a prophylactic / therapeutic agent for impairment of blood cell components such as reticulocytes and platelets, impaired hematopoiesis, and the like.

The nonapeptide used in the present invention is a known substance, and can be produced without difficulty by a peptide synthesis method in a liquid phase or a solid phase commonly used for peptide synthesis. Kaisho 54
-16425, USP.4301065). Alternatively, it can also be prepared by genetic engineering or cell engineering techniques.

The ester at the carboxyl group of asparagine at the C-terminus of the nonapeptide used in the present invention is a pharmaceutically acceptable ester of a carboxylic acid, and examples thereof include methyl ester, ethyl ester, propyl ester and isopropyl ester. Esters, n-butyl ester, isobutyl ester, tert-butyl ester, n-pentyl ester, isopentyl ester,
Neopentyl ester, tert-pentyl ester, n-
Hexyl ester, sec-hexyl ester, heptyl ester, octyl ester, sec-octyl ester, tert-octyl ester, nonyl ester, decyl ester, undecyl ester, dodecyl ester, tridecyl ester, tetradecyl ester, hexadecyl ester, octadecyl Ester, nonadecyl ester, eicosyl ester, cyclopentyl ester, cyclohexyl ester, cycloheptyl ester, cyclooctyl ester, allyl ester, isopropenyl ester, benzyl ester, o-, m-, or p-chlorobenzyl ester, o-, m- or p-fluorobenzyl ester, o-, m- or p-bromobenzyl ester, o-, m- or p-iodobenzyl ester, o-, m- or p -Methylbenzyl ester,
o-, m-, or p-ethylbenzyl ester, o-,
m- or p-isopropylbenzyl ester, cinnamyl ester, aminoethyl ester, o-, m- or p-aminobenzyl ester, o-, m- or p-
Nitrobenzyl ester, o-, m-, or p-methoxybenzyl ester, o-, m-, or p-ethoxybenzyl ester, o-, m-, or p-aminophenethyl ester, α-furfuryl ester, α-thienylmethyl ester, α-pyridylmethyl ester, α-pyridylethyl ester, piperidinomethyl ester, α-
Piperidyl methyl ester, morpholinoethyl ester, α-morpholinyl methyl ester and the like.

The amide at the carboxyl group of asparagine at the C-terminus of the nonapeptide used in the present invention is an amide of a pharmaceutically acceptable carboxylic acid, and examples thereof include amide, methylamide, ethylamide, and propylamide. , Isopropylamide, n-butylamide, isobutylamide, tert-butylamide, n
-Pentylamide, isopentylamide, neopentylamide, tert-pentylamide, n-hexylamide,
sec-hexylamide, heptylamide, octylamide, sec-octylamide, tert-octylamide, nonylamide, decylamide, undecylamide, dodecylamide, tridecylamide, tetradecylamide, hexadecylamide, octadecylamide, nonadecylamide, eico Silamide, cyclopentylamide, cyclohexylamide, cycloheptylamide, cyclooctylamide, allylamide, isopropenylamide, benzylamide, o-, m-, or p-chlorobenzylamide, o-, m-, or p-fluoro Benzylamide,
o-, m-, or p-bromobenzylamide, o-, m
-Or p-iodobenzylamide, o-, m- or p-methylbenzylamide, o-, m- or p-ethylbenzylamide, o-, m- or p-isopropylbenzylamide, cinnamyl Amide, aminoethylamide, o-, m-, or p-aminobenzylamide, o
-, M-, or p-nitrobenzylamide, o-, m
-Or p-methoxybenzylamide, o-, m- or p-ethoxybenzylamide, o-, m- or p-
Aminophenethylamide, α-furfurylamide, α-
Thienylmethylamide, α-pyridylmethylamide, α
-Pyridylethylamide, piperidinomethylamide, α
-Piperidylmethylamide, morpholinoethylamide,
α-morpholinylmethylamide, methoxycarbonyl-
(Α-mercaptomethyl) methylamide, ethoxycarbonyl- (α-mercaptomethyl) methylamide and the like.

Examples of the above-mentioned pharmaceutically acceptable salts include acid addition salts at the amino group and base salts at the carboxyl group of the nonapeptide. Examples of the acid addition salts include salts with organic acids or inorganic acids, and examples thereof include formic acid, acetic acid, propionic acid, trifluoroacetic acid, tartaric acid, fumaric acid, malic acid, maleic acid, and oxalic acid. Acid, salts with carboxylic acids such as naphthoic acid, salts with sulfonic acids such as methanesulfonic acid, p-toluenesulfonic acid, and naphthalenesulfonic acid, and salts with inorganic acids such as hydrochloric acid, sulfuric acid, nitric acid, and phosphoric acid. can give.

As the above-mentioned base salt, a salt with an inorganic base, that is, an alkali metal salt, an alkaline earth metal salt, an ammonium salt,
Alternatively, a salt with an organic base, that is, a salt with an amine is mentioned, and examples thereof include a lithium salt, a sodium salt, a potassium salt, a calcium salt, an ammonium salt, a triethylamine salt, an ethanolamine salt, a tris salt, and a dicyclohexylamine salt. And so on.

Examples of the metal in the compound of the compound and the metal include zinc, iron, magnesium, cobalt, nickel, chromium, gold, germanium, gallium, potassium, calcium, aluminum, copper, and manganese.
A valent and tetravalent metal is exemplified.

The preventive / therapeutic agent for disorders due to a decrease in blood cells such as leukocytes, reticulocytes, platelets, etc. according to the present invention is prepared using a commonly used formulation means according to the dosage form. That is, the active ingredient substance selected from the above nonapeptide and its ester or amide derivative or a salt thereof or a combination of each compound with a metal, as appropriate, a pharmaceutically acceptable carrier, excipient, diluent It is prepared into an appropriate dosage form using, for example. The dosage form can be various kinds of dosage forms suitable for external administration, oral administration, parenteral administration and the like.

The effects of the preventive / therapeutic agents for disorders due to a decrease in blood cell components such as leukocytes, reticulocytes, platelets, etc., and hematopoietic dysfunction according to the present invention have been confirmed by the following experiments.

That is, using a mammal such as a mouse, rat, guinea pig, dog, monkey, or Baboon, a cyclophosphine that induces suppression of bone marrow function, leukopenia, erythrocyte reduction, reticulocyte reduction, anemia, thrombocytopenia, etc., in these animals. Amide (cyclophospamide, hereinafter abbreviated as CPA),
Mitomycin C (hereinafter abbreviated as MMC), 5-fluorouracil (hereinafter abbreviated as 5-FU), adriamycin (adriamycin,
Abbreviated as ADM), daunomycin, achracinomycin, neocarzinostatin, vincristine, vinblastine, vindesine sulfate, nitromine, triethylenethiophosphoramide (TESPA), carbone (CQ),
Busulfan, Improsulfantsylate (864-
T), Nimustine hydrochloride (ACNU), Melphalan, Mitobronitol, Ranimustine (MCNU), Ifosfamide,
Methotrexate, 6-mercaptopurine, thioinosine, tegafur, carmofur, UFT, 5'-DFUR, cytarabine (Ara-C), Cyclo-C, mitoxantrone,
Actinomycin D, chromomycin A _3, L-asparaginase, Tamokishifuen, cisplatin, estramustine, etoposide, interferons such, a predetermined amount of an anticancer agent such as lentinan, administered orally or parenterally, a certain period time Blood test to determine white blood cell count,
Red blood cell count, reticulocyte count, platelet count, hemoglobin count,
Hematocrit value, average red blood cell volume, average red blood cell hemoglobin amount, average red blood cell hemoglobin concentration, etc. are measured, and at this time, a predetermined amount of the above nonapeptide is determined before or immediately after the administration of these anticancer agents, every day or every other day. Intra-peritoneal injection, subcutaneous injection, intramuscular injection, intravenous injection, oral administration and other various administration routes, blood test values, weight measurement values, etc. were compared and observed between the control group and the nonapeptide administration group . According to the results of this experiment, mice in the control group showed an increase in the degree of hematopoietic dysfunction as the dose of the anticancer drug was increased, and a decrease in the number of blood cells, worsened blood image, and suppression of weight gain were observed for a certain period of time. On the other hand, in the nonapeptide administration group, it was observed that the reduction of each blood cell was suppressed, the recovery of the blood cell count and the like to a normal range was promoted, and the suppression of weight gain was also slightly finished. Furthermore, the reduction of each blood cell of an animal to which a chemotherapeutic agent such as an antibacterial agent, an antiviral agent, or an immunosuppressant, or various drugs known to cause the reduction of each blood cell is reduced, the administration of the nonapeptide of the present invention. It was recognized that the recovery of each blood cell count to the normal range was promoted.

From the findings of these tests, the nonapeptide acts on hematopoietic organs such as bone marrow and spleen, and has an inhibitory effect on hematopoietic dysfunction or an effect of promoting recovery.As a result, disorders due to a decrease in blood cell components such as leukocytes, reticulocytes, and platelets, hematopoiesis It has been revealed that it can be used as a valuable drug as a prophylactic / therapeutic agent for dysfunction and the like.

In order to examine the toxicity of the active ingredient in the drug according to the present invention, 100 mg / kg of the active ingredient was administered daily to mice.
Although administered subcutaneously for 14 days, no abnormalities were observed in appearance. In addition, the active ingredient substance 30 mg / kg
Administered subcutaneously for days, both in appearance and in serum biochemistry,
No abnormalities were found in the results of the pathological dissection. Thus, the drug according to the present invention is a safe drug with extremely low toxicity, and can be administered for a long period of time.

Target animals to which the agent according to the present invention can be administered include, for example, humans and livestock such as cows, horses, pigs, sheep, goats, rabbits, dogs, cats, lions, elephants,
Giraffes, bears, gorillas, monkeys, chimpanzees and other zoos and other mammals, mice, rats, guinea pigs and other experimental animals, chickens and other poultry,
Examples include pet birds, reptiles, amphibians, and fish. The dose is usually 0.1 mg / kg body weight of these animals.
These may be administered in divided doses of, for example, once to six times a day at a dose of μg to 500 mg / day, and the dosage may be appropriately increased or decreased depending on the age, medical condition, and the like of the administration subject. Although the administration route is not particularly limited, it can be injected intravenously, intramuscularly, intradermally, or subcutaneously. In addition, by preparing a cartilage, it can be applied to the eye, oral cavity, nasal cavity, skin, etc., suppositories and jellies, eye drops, nasal drops, nasal and oral absorbents, aerosols, sprays, It can also be administered as an oral preparation or the like. In order to prevent rapid degradation or inactivation of the active ingredient substance in vivo, the active ingredient substance is converted into a suitable pharmaceutical ingredient, for example, an oily, fatty, harmless, harmless solid such as alcoholic, lecithin or the like. It can be made into a preparation using liposomes or the like, which is a liquid material or a suspension thereof, and can be a preparation having a long-lasting activity.

The drug according to the present invention can be administered together with other drugs, for example, a glutathione preparation, an inosine preparation, an adenine preparation, a cepharanthin preparation, and the like, which have a biological response modifier such as an immunostimulant and a leukopenia recovery effect. Can be added to the preparation to enhance the clinical effect as a mixture.

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

In the following examples, the nonapeptide is abbreviated as FTS.

Example 1 Injection vial formulation _{FTS · CH 3 COOH · 2H 2} O and (Mitsui Seiyaku Kogyo Co., Ltd.) 1 mg was dissolved in distilled water and filtered Jokinro, filled into vials and lyophilized.

EXAMPLE 2 Injectable ampoules preparation _{FTS · CH 3 COOH · 2H 2} O ( manufactured by Mitsui Chemical Industry Co., Ltd.) 5 mg was dissolved in physiological saline, filtered Jokinro, filled into ampoules.

Example 3 Injection for Subcutaneous Injection FTS · CH ₃ COOH · 2H ₂ O (manufactured by Mitsui Pharmaceutical Co., Ltd.) was suspended at 2 mg per unit dose in a 2% carboxymethylcellulose PBS (phosphate buffered saline) solution. Lipomal composed of soy phosphatide (Huhtamaki Oy / Leiras Pharmaceuti
cals) or Intral, an oil-in-water emulsion for intravenous use
It was mixed with ipid (Cutter Laboratories). Lipomal
When using FTS, the FTS-dissolved PBS solution and Lipomal were mixed in equal amounts.

When using Intralipid, 2.5 ml of FTS-dissolved PBS solution, T
0.1 ml of ween 80 (manufactured by Sigma Chemicals) and Intralipid
4.6 ml were mixed.

Example 4 Liposomal Formulations There are three types of liposome formulations having different charges, and these are further classified into four types in terms of structure.

Charges are neutral, positive and negative, and are structurally multilayered liposomes (MLV, mulitlamellarvesicle) and small unilamellar liposomes (SUV, small unilamellar vesicl)
e) and large unilamellar liposomes (LUV, large unil
amellar vesicle) and several films with a structure similar to LUV (REV, reverse-phase evaporation vesi)
cle) are known.

FTS-encapsulated neutral-charge liposomes Phosphatidylcholines, phospholipids such as sphingomyelin, and a cholesterol chloroform solution are mixed in a molar ratio of 2: 1, 4: 1, or 1: 1. Distilled under reduced pressure, 1 / 100-1 / 100
A solution of 0 equivalent of FTS in PBS (phosphate buffered saline) was added and thoroughly mixed with a Vortex miver to obtain MLV.

Further, SUV was obtained by sonication at a phase transition temperature (Tc) of the phospholipid or higher.

An aqueous calcium chloride solution was added to the obtained SUV, and the mixture was incubated at 37 ° C. for 1 hour to fuse. Then, EDTA was added, and the mixture was incubated at 37 ° C. for 30 minutes to remove Ca ^++.
V was obtained.

The method for preparing REV is as follows. That is, after the solvent was distilled off from the chloroform solution of lipid under reduced pressure, diethyl ether was added and dissolved in a sufficient amount to give PBS of FTS.
The solution was added and sonicated to form a uniform single phase solution. After the obtained solution was concentrated under reduced pressure at room temperature, a PBS solution was added and thoroughly mixed with a Vortex mixer to obtain PEV.

FTS-encapsulated positively charged liposomes The preparation method is the same as that for the neutrally charged liposomes described above, except for the components of the lipid.

Phosphatidylcholines, phospholipids such as sphingomyelin, cholesterol, and positively charged higher aliphatic amines such as stearylamine in a molar ratio of 7: 2: 1 or 4: 1: 1.
And the mixture was used as a lipid component, and FTS was encapsulated in the same manner.

FTS-encapsulated negatively charged liposomes, phosphatidylcholines, phospholipids such as sphingomyelin, cholesterol and dicetyl phosphate,
Negatively charged higher aliphatic esters such as sulfatide were mixed at a molar ratio of 7: 2: 1 or 4: 1: 1 to obtain lipid components, and FTS was encapsulated in the same manner.

Example 5 Cartilage agent FTS · CH ₃ COOH · 2H ₂ O (manufactured by Mitsui Pharmaceutical Co., Ltd.) (2 mg) was dissolved in purified water. Next, 25 g of white petrolatum, 20 g of stearyl alcohol, 4 g of HCO-60 and 1 g of glyceryl monostearate were mixed, mixed and prepared in advance, 12 g of propylene glycol, 0.1 g of methyl parahydroxybenzoate, 0.1 g of an aqueous solution of 0.1 g of propyl paraoxybenzoate (FTS Was added and thoroughly mixed to obtain an emulsion, and the mixture was cooled and solidified to continue the mixing operation.

Example 6 suppository _{FTS · CH 3 COOH · 2H 2} O ( manufactured by Mitsui Chemical Industry Co., Ltd.) 10 mg was dispersed in prewarmed Hadofuatsuto were a total volume of 2g.

Example 7 Nasal Capsule 0.05 mg of FTS was dissolved in 29.95 mg of Miglyol 812 neutral oil (Dynamite Nobel) under aseptic conditions. This solution was filled into a conventional unit-dose dispenser, which was mounted on a drive capsule before use.

Example 8 Nasal Drops The following amounts of sodium dihydrogen phosphate, disodium hydrogen phosphate, sodium chloride and EDTA-disodium salt were dissolved in distilled water at room temperature. Dissolve FTS in this solution,
Spent by membrane filter.

FTS 0.10mg sodium monohydrogen phosphate ・ 2H ₂ O 0.30mg sodium dihydrogen phosphate ・ 12H ₂ O 10.10mg benzalkonium chloride 0.10mg ethylenediaminetetraacetic acid-disodium salt (EDTA)
The 50mg Sodium chloride 4.50mg distilled water 987.60Mg pH value 5.0 ± 0.3 Example 9 nasal spray formulations _{FTS · CH 3 COOH · 2H 2} O ( manufactured by Mitsui Chemical Industry Co., Ltd.) 2 mg were suspended in hydroxypropylcellulose or hydroxypropylmethylcellulose The resulting mixture was used as a spray agent by a spray formulation machine.

Hereinafter, examples of pharmacological experiments and toxicity experiments on the drug of the present invention will be described.

Experimental Example 1 Effect on mice administered orally with mitomycin (MMC) Female 7-week-old ICR mice (Clea 9 Japan was prepared in groups of 3 each, and the following groups were treated in total: 4 groups. The first group was 0.1 ml. Physiological saline was intraperitoneally administered to the mice for four consecutive days, and the second group contained FTS, CH ₃ COOH, and 2H ₂ O used in Example 1 (hereinafter abbreviated as FTS substance in the description of each experimental example). Mouse to 1
It was administered intraperitoneally 100 μg once a day for 4 consecutive days. Third
The group was orally administered 5 mg / kg of MMC once a day on the test day. The fourth group received the FTS substance once a day from 2 days before MMC administration for 4 consecutive days.
Intra-peritoneally 100 μg each, MMC was orally administered once at 5 mg / kg on the third day after the start of administration of the FTS substance. Immediately before administration of the drug, blood was collected daily from the ocular vein, and white blood cells were counted after Turk staining.
Table 1 shows the results. As is clear from Table 1, the FTS substance was observed to prevent and prevent the transient leukopenia in peripheral blood on the day after administration by the administration of MMC, and the hematopoietic dysfunction caused by the administration of the anticancer drug was observed. Of FTS substances as prophylactic / therapeutic agents against cancer was clarified.

Experimental Example 2 Effect on Mice Administered Orally with 5-Fluorouracil (5-FU) Simultaneously with Experimental Example 1. Third of Experimental Example 1
The group to which 5-FU was orally administered once at 50 mg / kg instead of the group was designated as a fifth group. Also, 2 days before 5-FU administration and 4 consecutive days,
The FTS substance was intraperitoneally administered at a dose of 100 μg once a day.
The group to which oral administration was performed was designated as a sixth group. The same inspection as in Experimental Example 1 was performed, and the obtained results are shown in Table 2. As is evident from Table 2, the FTS substance was found to prevent and prevent leukopenia caused by 5-FU administration.

Experimental Example 3 Effect on MMC Intraperitoneally Administered Mice Six female 5-week-old ICR / JCL mice were prepared per group, and the following four groups were treated in total. In the first group, 0.1 ml of saline was intraperitoneally administered to mice for 12 consecutive days. In the second group, the FTS substance was subcutaneously administered to mice 100 μg once a day for 12 consecutive days. In the third group, MMC was intraperitoneally administered once a day at 20 μg / 0.1 ml for 10 days from the 0th to the 9th day of the test. In the fourth group, FTS was subcutaneously administered at a dose of 100 μg once a day for 2 days from the day before MMC administration for 12 consecutive days. From 3 days after the start of FST substance administration, MMC was administered once a day at 20 μg / 0.1 ml, intraperitoneally. Was administered. After administration of anticancer drug,
Blood was collected from the ocular vein every other day and leukocytes were counted. Table 3 shows the results
Shown in

The white blood cell count in the peripheral blood of the group to which the FTS substance alone was administered was not larger than the white blood cell count of the group to which saline was administered, and was within the normal range. In the MMC-only administration group, the white blood cell count began to decrease 3 days after the initial administration, reaching about 40% of the maximum normal range. The decreasing tendency was observed for one week or more after the end of MMC administration. On the other hand, the white blood cell count in the MMC + FTS substance combination administration group was
It was confirmed that the ratio was maintained at about 60 to 80% of the group administered with saline, and that the recovery to the normal range after the administration of the anticancer agent was promoted.

Experimental Example 4 Effect on 5-FU intraperitoneally administered mouse Simultaneously as in Experimental Example 3. Third of Experimental Example 3
The fifth group was a group in which 5-FU was intraperitoneally administered once a day at 60 μg / 0.1 ml for 10 days from the 0th to the 9th day of the test instead of the group. Also, a group in which 100 μg of the FTS substance was subcutaneously administered once a day from 2 days before the administration of 5-FU to 12 consecutive days, and 60 μg / 0.1 ml of 5-FU was intraperitoneally administered for 10 days from the third day of the administration of the FTS substance. Was designated as a sixth group. The same inspection as in Experimental Example 3 was performed, and the obtained results are shown in Table 4. In the 5-FU alone administration group, the leukocyte count began to decrease 3 days after the initial administration, reaching approximately 37% of the maximum normal range. The decreasing tendency was observed for one week or more even after the end of 5-FU administration. On the other hand, the leukocyte count in the group administered with the 5-FU + FTS substance was maintained at about 60 to 90% of that in the group administered with saline, and it was confirmed that the recovery to the normal range after the administration of the anticancer agent was promoted.

Experimental Example 5 Effect on mice intraperitoneally administered with cyclophosphamide (CPA) Ten groups of male 8-week-old C3H / HeN mice were prepared.
In the group, 0.2 ml of the raw food was subcutaneously administered to the mice subcutaneously for 12 days from test day 2 to test day 9. In the second group, the FTS substance was subcutaneously administered to the mice once a day at 100 μg / 0.2 ml for 12 consecutive days. The third group received 400 mg / kg of CPA intraperitoneally once on the day of the test. In the fourth group, a dosing schedule combining the second group and the third group was administered. Group 5: 200mg / k CPA
g, was administered intraperitoneally once on the day of the test. The sixth group performed a dosing schedule combining the second group and the fifth group. Group 7 received CPA at 100 mg / kg intraperitoneally once on the day of the test. Group 8 was tested with a dosing schedule combining groups 2 and 7. Test day (0, 3,
On days 5, 7, 10, and 14), the arteries and veins in the armpit of the mouse were cut off, blood was collected, blood tests were performed, and the white blood cell count (WBC) and red blood cell count (R
BC), reticulocyte count (RET), platelet count (PLT), hemoglobin count (HGB), hematocrit value (HCT), average red blood cell volume, average red blood cell hemoglobin amount, average red blood cell hemoglobin concentration, and the like. In each of the CPA 400 mg / kg, 200 mg / kg, and 100 mg / kg single dose groups, WBC
Although it decreased sharply on the fifth and seventh days, the decrease in WBC was suppressed in the group administered with the FTS substance. For example, the mean value of WBC ± SD of 45.2 ± 5.0 (× 10 ² / mm ³ ) in the group administered with the saline on the ^third day is 48.0 ± 4.7, and the CPA 40
0mg / kg single administration group was 11.6 ± 2.1, CPA + FTS substance combined administration group was 17.0 ± 2.0, and
In contrast to 41.8 ± 6.5, the FTS substance alone administration group was 38.0 ± 6.1, C
The PA 400 mg / kg single administration group was 4.2 ± 1.0, the CPA + FTS combination administration group was 6.4 ± 0.3, and the FTS substance combination group showed a decrease in the reduction of WBC. A promoting effect was seen.

On the other hand, RET decreased in each CPA administration group at the bottom on the fifth day, but on the seventh day, the CPA + FTS substance combination administration group showed a CPA 20
In the case of 0 mg / kg and 100 mg / kg administration, it recovered to the normal range. For example saline administration group on day 7 whereas a _{^{52.6 ± 7.3 (0/00)}} , FTS substance alone group 68.8 ± 12.0, CPA 400mg / k
g 27.2 ± 5.0 for single administration group, 4 for CPA + FTS substance combination administration group
0.8 ± 7.2. The FTS substance combination administration group suppressed the decrease in RET and promoted the recovery.

In the case of PLT, CPA 400 mg / kg, 200
In the mg / kg administration group, it decreased, but recovered to the normal range on the 10th day. For example, on the 5th day, 933 ± 2
6.0 (× 10 ³ / mm ³ ), whereas the FTS substance single administration group was 804 ± 162, the CPA 400 mg / kg single administration group was 357 ± 82, CPA +
The FTS substance combination administration group was 580 ± 49, and the raw food group on day 7 was 724 ± 60, whereas the FTS substance alone administration group was 70 ± 49.
7 ± 172, 214 ± 73 for CPA 400mg / kg single dose group, CPA + FTS
The value of the group administered with the substance was 540 ± 87. The FTS substance combination administration group suppressed the decrease in PLT and accelerated the recovery.

On the other hand, it was revealed that the FTS substance-combined administration group also suppressed the decrease in other blood parameters of blood tests, that is, RBC, HGB, HCT, etc., and accelerated their recovery. It was also revealed that the group administered with the FTS substance had an effect of preventing the suppression of weight gain in the group administered with CPA alone.

Experimental Example 6 Toxicity test Five groups of 5 week-old ddY male mice were treated with 50
g / kg and 100 mg / kg were administered subcutaneously for 14 days each day, but no toxicity was observed.

Experimental Example 7 Toxicity test 5 groups of 10-week-old wister rats were treated with active substance
Subcutaneous administration of 30 mg / kg for 21 consecutive days showed no toxicity.

As described above, in addition to leukopenia-recovery agents that have been used only slightly in the past, there are almost no effective agents, disorders due to a decrease in blood cells such as leukocytes, reticulocytes, and platelets, and hematopoiesis. In the medical field for protecting dysfunction, etc., the agent according to the present invention activates the immune system or the biological defense system, thereby causing disorders due to a decrease in blood cells such as leukocytes, reticulocytes, platelets, hematopoietic dysfunction, etc. It is a breakthrough in that it provides excellent preventive and therapeutic effects. In addition, the nonapeptide (FTS) used in the present invention is a peptide derived from an animal and is a natural substance. Therefore, unlike FTS analogs (analogs) having a similar amino acid sequence, it is completely nontoxic in living organisms, There are no problems such as anaflexism.
Therefore, the drug according to the present invention is safe and useful for humans,
It can be used as a medicine for animals.

──────────────────────────────────────────────────続 き Continued on the front page (56) References Patent 2655343 (JP, B2) (58) Fields investigated (Int. Cl. ⁷ , DB name) A61K 38/22 CA (STN) REGISTRY (STN)

Claims (1)

(57) [Claims] (1) the following amino acid sequence: Or an ester, an amide or a pharmaceutically acceptable salt of each of these compounds at the carboxyl group of asparagine at the C-terminus or a pharmaceutically acceptable salt of each of these compounds or a metal-bound compound as an active ingredient. An agent for preventing or treating a blood cell depletion disorder caused by administration of a chemotherapeutic agent.