JPH11286455A - Medicine for osteomyelodysplasia syndrome - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain the subject medicine for improving hematopenia accompanied by osteomyelodysplasia syndrome by adding a matrix metalloprotease inhibitor, a TNF-α transferase inhibitor and a Fas ligand solubilization inhibitor as active ingredients. SOLUTION: This medicine contains a matrix metalloprotease inhibitor, a TNF-α transferase inhibitor and/or a Fas ligand solubilization inhibitor for example, [4-(N-hydroxyamino)-2(R)-isobutyl-3-methylsuccinyl]-L-3-(5,6,7,8- tetrahydro-1-naphthyl)alanine-N-methylamide} as active ingredients. The medicine is preferably used for hypocytosis recognized in refractory anemia, sideroblastic refractory anemia, refractory anemia accompanying the increase of gemmules, etc. The medicine is orally or parenterally administered into patients. The medicine is administered at a daily dose of 1-600 mg converted to the active ingredient with one, two, three or four portions.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION Matrix metalloprotease (hereinafter referred to as MMP) is an enzyme that degrades a protein component constituting a connective tissue matrix, and a compound that inhibits MMP is used for treating connective tissue diseases such as arthritis and osteoporosis. It is expected to be developed as an agent (MMP inhibitor).

On the other hand, it has been revealed that metalloproteases similar to MMPs are involved in various processes in vivo.

For example, tumor necrosis factor (TNF-α) (17
kD) is a metalloprotease (TNF-α convertase, TAC) derived from a precursor (26 kD) existing on the cell surface.
E) was found to be cut out and produced [Mohler et al., Nature, 370, 218-220 (1994)].

Further, regarding apoptosis, a ligand of Fas (FasL) is a certain metalloprotease (FsL).
A mechanism has been proposed in which soluble FasL is solubilized by an as ligand solubilizing enzyme, and cell death is caused as a result of binding to Fas [Kayagaki
J. Exp. Med., 182, 1777-1783 (1995)].

On the other hand, myelodysplastic syndrome (hereinafter, MDS)
This is caused by impaired maturation of bone marrow hematopoietic cells caused by impaired hematopoietic stem cell levels of unknown cause, resulting in cytopenias due to myelodysplasia and bone marrow hypoplasia It is a disease. The prognosis of MDS is very poor and the average survival rate is about two years from diagnosis.

[0006] The present invention relates to the above MMP inhibitor, TACE
The present invention relates to a therapeutic agent for MDS comprising an inhibitor and / or an inhibitor of a Fas ligand solubilizing enzyme as an active ingredient.

[0007]

2. Description of the Related Art The treatment of MDS involves anemia associated with red blood cell reduction,
For the purpose of protecting against bleeding tendency associated with thrombocytopenia or infection associated with granulocytopenia, treatments aimed at improving cytopenia have been performed. Administration of prednisolone, anabolic hormones, vitamin D3, hematopoietic factors (cytokines), etc. has been attempted for the purpose of improving cytopenia, but little effect has been observed (Yoshina Kodera, Hidehiko Saito, Manual for Hematopoietic Stem Cell Transplantation) , Japan Medical Museum, 1995,
30 pages).

Recently, attention has been paid to the idea that hematopoietic disorders of MDS are caused by apoptosis of bone marrow cells. In fact, it has been reported that Fas ligand and its receptor that cause apoptosis are increased in bone marrow cells of MDS patients [Blood, 88 (3), 1122 (1996)]. In addition, it has been reported that cytokines known to induce apoptosis, such as TNF-α and interleukin (IL1β), are increased in MDS patients [Raza et al., Int.
J. Hematol., 63, 265 (1996)]. Against this background, studies have been made to use an inhibitor of apoptosis for MDS treatment, and Raza et al. Have observed a certain effect by administering pentoxifylline (a signaling inhibitor of the cytokine) to MDS patients [ Raza et al., Blood, 88 (Supple), 231
0 (1996)]. It has also been proposed to use a polypeptide having an apoptosis-suppressing activity as a therapeutic agent for MDS [PCT International Publication No. WO97 / 6255].

On the other hand, since MMP inhibitors suppress the solubilization of Fas ligand, diseases caused by excessive T cell activation, such as hepatitis, graft-versus-host reaction (GVH)
D) Therapeutic use of MMP inhibitors (Fas ligand solubilization inhibitor) for autoimmune diseases and the like has been filed (PCT International Publication WO97 / 9066). Also,
Some metalloproteases have also been filed for use as TACE inhibitors, ie, therapeutic agents for diseases mediated by excess TNF-α (eg, inflammation, acute infections, shock states, GVHD, etc.) [PCT International Publication WO94 / 10990]. However, MDS
There are no known metalloprotease inhibitors that suppress apoptosis of bone marrow cells in patients.

[0010]

It is known that bone marrow cells of MDS patients are fragile and easily killed compared to bone marrow cells of healthy persons. That is, the bone marrow cells of the MDS patient are taken out
After culturing for 24 hours, it was observed that about 60% of the cells undergo apoptosis and die, which is considered to be a cause of the cytopenia in MDS. An object of the present invention is to provide MDS
An object of the present invention is to find a drug that improves the cytopenia associated with MDS and to provide a therapeutic agent for MDS.

[0011]

As a result of various studies, the present inventors have found that the following compounds strongly suppress apoptosis of cultured bone marrow cells of MDS patients, and have completed the present invention.

[0012]

BEST MODE FOR CARRYING OUT THE INVENTION The present invention relates to an MMP inhibitor, TAC
An E inhibitor and / or a Fas ligand solubilization inhibitor is used as an active ingredient. Examples of the MMP inhibitor, TACE inhibitor or Fas ligand solubilization inhibitor that can be used in the present invention include, for example, metalloprotease inhibitors described in the following publications.

Japanese Unexamined Patent Publication No. 7-101925 US Pat. No. 4,743,587 International Publication WO92 / 09556 US Pat. No. WO93 / 09090 US Pat. No. WO93 / 24449 US Pat. No. WO95 / 04033 European Patent Publication 489579 US Pat. No. 497192, No. 575844, International Publication WO90 / 05719, No. WO94 / 21625, No. WO94 / 24140, No. WO94 / 02447, No. 6-145148, No. 7-157470, No. WO97 / 20824. JP 97/18194 JP WO 96/33172 JP WO 96/15096 JP WO 95/13289 JP 9-188431 JP 7-291938 JP 6-256293 Particularly preferable is a compound of the following.

[4- (N-hydroxyamino) -2
(R) -isobutyl-3 (S) -methylsuccinyl]-
L-phenylglycine-N-methylamide (Japanese Unexamined Patent Publication No.
No. 101925) [4- (N-hydroxyamino) -2 (R) -isobutyl-3 (S)-(2-thienylthiomethyl) succinyl] -L-phenylalanine-N-methylamide (WO90 / 05719) ) 3 (S) -N-hydroxy-4- (4-((pyrido-4
- yl) oxy) benzenesulfonyl) -2,2-dimethyl - tetrahydro-2H-1,4-thiazine-3-carboxamide (International Publication WO97 / No. ^{20824) N 2 - [3 (} S) - hydroxy-4- (N-hydroxyamino) -2 (R) -isobutylsuccinyl] -L-te
rt-leucine-N ¹ -methylamide (WO WO
No. 94/02447) In the MDS therapeutic agent of the present invention, a FasL solubilization inhibitor may be used as an active ingredient. FasL solubilization inhibitors include, for example, PCT International Publication WO97
/ 9066 can be used.

Among them, the following compounds are preferred.

[4- (N-hydroxyamino) -2
(R) -isobutyl-3-methylsuccinyl] -L-3
-(5,6,7,8-tetrahydro-1-naphthyl) alanine-N-methylamide [4- (N-hydroxyamino) -2 (R) -phenylpropylsuccinyl] -L-phenylglycine-N-methylamide MMP inhibitors, TACE inhibitors or Fa
Any of the sL solubilization inhibitors can be manufactured by the methods described in the respective publications, and any one of the agents can be used alone or in combination of two or more.

MDS is refractory anemia (RA), refractory anemia with sideroblasts (RARS), refractory anemia with increased blasts (RAEB), RAEB becoming leukemia
(RAEB-t), chronic myelomonocytic leukemia (CMMo)
Although the LDS is classified into the five disease groups of L), the therapeutic agent for MDS of the present invention can be suitably used especially for cytopenia observed in RA, RARS, RAEB and the like.

The therapeutic agent for MDS of the present invention is administered orally or parenterally to a patient. The dosage depends on the target syndrome,
Although it varies depending on the administration method, patient age, body weight, symptoms, etc., it is generally 1 to 6 in terms of the active ingredient, ie, MMP inhibitor, TACE inhibitor or Fas ligand solubilization inhibitor.
A suitable range is 00 mg / day, which is administered once a day or divided into two to four times a day.

As the dosage form, tablets, granules, fine granules, capsules, injections, liposomes and the like are used depending on the administration form of oral or parenteral administration.

Such a preparation is manufactured by a conventional method, and tablets, granules and fine granules are used as active ingredients such as MMP inhibitors,
Disintegration of TACE inhibitors and / or Fas ligand solubilizing inhibitors and usual pharmaceutical additives such as lactose, synthetic aluminum silicate, glucose, excipients such as mannitol, crystalline cellulose, starch, carboxymethylcellulose, sodium alginate, etc. Agent, magnesium stearate, a lubricant such as talc, or a binder such as hydroxymethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone and the like. It is manufactured by filling into capsules.

Injectable preparations include an active ingredient, an MMP inhibitor,
A TACE inhibitor and / or a Fas ligand solubilizing inhibitor are dissolved or suspended in sterile water, and to this is added an isotonic agent such as mannitol, sodium chloride, glucose, sorbitol, glycerol, xylitol, fructose, maltose, mannose, etc. If necessary, a stabilizer such as sodium sulfite and albumin and a preservative such as benzyl alcohol are added, and the mixture is aseptically encapsulated in ampoules or vials.

[0022]

EFFECT OF THE INVENTION The therapeutic agent for MDS of the present invention strongly inhibits apoptosis of cultured bone marrow cells (see the following Test Examples), and is particularly useful as a therapeutic agent for MDS since no toxicity is observed. <Test Example> 1. Test compound Compound a: [4- (N-hydroxyamino) -2 (R)
-Isobutyl-3 (S) -methylsuccinyl] -L-phenylglycine-N-methylamide (JP-A-7-101
No. 925) Compound b: [4- (N-hydroxyamino) -2 (R)
-Isobutyl-3-methylsuccinyl] -L-3-
(5,6,7,8-tetrahydro-1-naphthyl) alanine-N-methylamide (PCT International Publication WO97
No./9066) Compound c: [4- (N-hydroxyamino) -2 (R)
-Phenylpropylsuccinyl] -L-phenylglycine-N-methylamide (PCT International Publication WO97 /
No. 9066) Compound d: N ^2- [3 (S) -hydroxy-4- (N-
[Hydroxyamino) -2 (R) -isobutylsuccinyl] -L-tert-leucine-N ¹ -methylamide (WO 94/02447) Compound e: 3 (S) -N-hydroxy-4- (4-
((Pyrid-4-yl) oxy) benzenesulfonyl)
-2,2-dimethyl-tetrahydro-2H-1,4-thiazine-3-carboxamide (International Publication WO97 /
No. 20824)

2. Test Method Bone marrow fluid was collected from the bone marrow of an MDS patient using a bone marrow puncture needle in the presence of heparin, bone marrow nucleated cells were separated by specific gravity centrifugation, and RPMI-1640 culture medium supplemented with 10% fetal bovine serum (antibiotics) (Containing penicillin and streptomycin) at a concentration of 1 × 10 ⁵ / mL. The test compound is dissolved in dimethylsulfoxide (DMSO) to a concentration of 10 mM and diluted with the culture solution to give a final concentration of 10 or 2
It was adjusted to a concentration of 0 μM and added. The culture was performed using a carbon dioxide incubator (5% carbon dioxide, 95% indoor air). After 18 hours of culture, a cytospin smear was prepared using a centrifuge for floating cells, and after sufficiently dried,
Boehringer Mannheim Cell Death Detection Kit (in situ
Apoptotic cells were stained by an end labeling method (in situ end labeling method) using a u cell death detection kit, and apoptotic cells were determined under a light microscope. In all experiments, at least 500 or more cells were observed, and the ratio (%) of apoptotic cells in the test compound-added group to the drug-free group was calculated. The inhibitory effect on apoptosis induction was determined by paired t-test (paired t-test).
ed t test). 3. Test results Table 1 shows the test results.

[0024]

[Table 1]

[0025]

The present invention will be described more specifically with reference to the following examples. Example 1 Preparation of Tablets [4- (N-Hydroxyamino) -2
(R) -isobutyl-3 (S) -methylsuccinyl]-
L-phenylglycine-N-methylamide (compound a)
A tablet containing 100 mg is obtained. [Operation] The main drug, corn starch and microcrystalline cellulose are mixed, and hydroxypropylcellulose dissolved in 50 parts by weight of water is added thereto and kneaded sufficiently. The kneaded product is passed through a sieve, granulated on granules, and dried. Then, the obtained granules are mixed with magnesium stearate and compressed into 250 mg tablets.

[0026] As the manufacturing example 2 below ^{granules, N 2 - [3 (S} ) - hydroxy-4-(N
-Hydroxyamino) -2 (R) -isobutylsuccinyl] -L-tert-leucine-N ¹ -methylamide (compound b) 200 mg is obtained. [Operation] A main drug, lactose and corn starch are mixed, and hydroxypropylcellulose dissolved in 120 parts by weight of water is added and kneaded sufficiently. The kneaded product is passed through a 20-mesh sieve, granulated, dried and sized, and then 500 mg is packaged to obtain granules.

[0027] As preparation Example 3 below capsules, N ² in 1 capsule - [3 (S) - hydroxy-4-(N-hydroxyamino) -2 (R) - isobutyl succinyl] -L-tert A capsule containing 100 mg of -leucine-N ¹ -methylamide (compound b) is obtained. [Operation] Thoroughly mix the above components, and mix 2
A capsule is obtained by filling the capsule with the amount of 00 mg.

Example 4 Preparation of Injection [4- (N-hydroxyamino) -2 (R) -isobutyl-3 (S) -methylsuccinyl] -L-phenylglycine-N-methylamide 0.5 part by weight and Distilled water for injection was added to 5 parts by weight of sorbitol to dissolve the mixture,
The solution is made 0 parts by weight, and this aqueous solution is filtered with a membrane filter. The filtrate was filled into ampoules purged with nitrogen in an amount of 5 g, sealed, sterilized at 120 ° C. for 15 minutes, and [4- (N-hydroxyamino) -2 (R) -isobutyl-3 (S) was contained in one ampule. -Methylsuccinyl] -L-phenylglycine-N-methylamide in 25 mg is obtained.

 ──────────────────────────────────────────────────続 き Continued on the front page (72) Inventor Mitsuo Arimura 4-32-18 Shimoarata, Kagoshima City (1F Morinaga)

Claims (4)

[Claims] (1) a matrix metalloprotease inhibitor,
A therapeutic drug for myelodysplastic syndrome, comprising a TNF-α convertase inhibitor and / or a Fas ligand solubilization inhibitor as an active ingredient. 2. The therapeutic agent for myelodysplastic syndrome according to claim 1, which comprises a matrix metalloprotease inhibitor as an active ingredient. 3. The therapeutic agent for myelodysplastic syndrome according to claim 1, which comprises a TNF-α convertase inhibitor as an active ingredient. 4. The therapeutic agent for myelodysplastic syndrome according to claim 1, which comprises a solubilizing inhibitor of Fas ligand as an active ingredient.