JP2002047281A - Telomerase inhibitor - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a compound having an inhibitory action on telomerase and a telomerase inhibitor, a medicine and an antitumor medicine comprising the compound as an active ingredient. SOLUTION: This compound 1 is represented by general formula (I). The telomerase inhibitor, the medicine and the antitumor medicine comprise the compound 1, its physiological acceptable salt, its hydrate or its solvate as an active ingredient.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a telomerase inhibitor. The present invention also relates to a medicament useful as an antitumor agent containing the telomerase inhibitor.

[0002]

2. Description of the Related Art In recent years, some cancers typified by Hodgkin's disease have become curable, and effective treatment methods have been developed for many other cancer types. In addition, the mechanism of malignant transformation of cells, growth factors and their receptors, intracellular signaling systems,
There have been many reports on the role of oncogenes and tumor suppressor genes. However, as the understanding of cancer deepens, it is becoming clear that the mechanism of malignancy that leads to the formation of cancer is not common to individual cancers, and that cancer is a very diverse disease. It is becoming clear that the complexity of the combination of malignant factors in each cancer is a major barrier to cancer treatment.

On the other hand, recently, the relationship between the aging of normal cells and the mechanism of immortalization of cancer cells has been elucidated at the genetic level. The chromosomal DNA of eukaryotic cells is linear, and its terminal portion consists of a simple repeating sequence called telomere. It has long been known that this telomere plays an important role in chromosome structure and function [Moyzis RK et al., P.
roc Natl Acad Sci USA, 85, p. 6622 (1988)], gradually loses telomeres as the cells divide, and after a certain number of divisions loses the ability to proliferate, resulting in senescence of the cells,
It was suggested that telomerase, an enzyme that synthesizes and adds telomeres, plays an important role in immortalizing cancer cells. [Kim
NW et al., Science, 266, p. 2011 (1994)].

[0004] It is known that most cancer cells have telomerase activity, and it is thought that this allows antagonism of telomere shortening accompanying cell division to allow unlimited growth. On the other hand, many normal somatic cells do not have telomerase activity, and telomere shortening accompanying cell division and cell senescence due to the shortening are observed. From these facts, it is considered that an inhibitor of telomerase activity can selectively inhibit the infinite growth of cancer cells, and is an antitumor agent applicable to a wider variety of cancer types (Egorov E.
E. et al., Biochemistry, 62, p. 1296 (1997)].

[0005]

An object of the present invention is to provide a telomerase inhibitor. Another object of the present invention is to provide a medicament useful as an antitumor agent or the like, which contains a substance having a telomere inhibitory action as an active ingredient.

[0006]

The present inventors have focused on the fact that microorganisms produce bioactive substances such as antibiotics,
Numerous samples were collected from nature, and intensive studies were conducted on various types of cultures separated therefrom to find substances having a telomerase inhibitory action.

[0007] As a result, it has been found that a novel substance having a telomerase inhibitory action is contained in a culture of a strain belonging to the order of the Moniliaria imperfect fungi.
The present inventors have clarified the structure of the substance and completed the present invention.

That is, the present invention relates to compound 1, represented by the following general formula (I), a physiologically acceptable salt thereof, a hydrate thereof, or a solvate thereof.

[0009]

Embedded image The present invention is also a telomerase inhibitor comprising the above compound 1, a physiologically acceptable salt thereof, a hydrate thereof or a solvate thereof as an active ingredient.

Further, the present invention is a medicament comprising the above compound 1, a physiologically acceptable salt thereof, a hydrate thereof or a solvate thereof as an active ingredient.

Further, the present invention is an antitumor medicament comprising the above compound 1, a physiologically acceptable salt thereof, a hydrate thereof or a solvate thereof as an active ingredient.

[0012] The present invention relates to the genus Epicoccum.
A method for producing Compound 1 comprising culturing a microorganism having the ability to produce Compound 1 and collecting Compound 1 from the resulting culture.

In the method for producing compound 1 of the present invention, the microorganism capable of producing the compound 1 is epicoccam.
Perpura Sense (Epicoccum purpurascens) D86
Preferably, the strain is 46 strains (FERM P-17859).

[0014]

DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will be described below in detail.

When the compound 1 is contained as the telomerase inhibitor, medicament or antitumor medicament of the present invention, its steric structure including the pentavinylene group moiety (-(CH = CH) _5- ) in the formula is cis-form, Although it is possible to include both trans types, it is also possible to include only the cis type or only the trans type, and there is no particular limitation.

The compound 1 of the present invention has various isomers (optical isomers and diastereoisomers) because of the presence of an asymmetric carbon atom in the molecule. Compound 1 of the present invention
Includes all of these isomers and a mixture of these isomers, and the steric structure is not particularly limited. In this case as well, when compound 1 is contained as the telomerase inhibitor, drug or antitumor drug of the present invention, it can be used as a mixture of isomers, but is preferably used alone and is not particularly limited.

The compound represented by the general formula (I) of the present invention is
By leaving in the air or recrystallizing,
Absorbs water or solvents and may absorb water or become hydrates or solvates.
Solvates are also included in the present invention.

According to the present invention, compound 1 can be obtained by culturing a microorganism capable of producing compound 1 and collecting the compound from the resulting culture. The microorganism used for the production of Compound 1 is not particularly limited as long as it is a microorganism capable of producing Compound 1, and is, for example, classified into the incomplete fungal subphylum fungal web having the ability to produce Compound 1. Molds can be used. It is preferably a genus Epicoccum belonging to the order of the subphylum Mycorrhizal subfamily Monilidia, and particularly preferably Epicoccum purpurascens D8.
646 strains.

Usually, Compound 1 can be efficiently produced by collecting Compound 1 from a culture obtained by culturing the producing bacterium by separating and purifying the Compound 1 by a conventional method. Chemical methods such as synthetic or semi-synthetic,
Alternatively, it can be produced by a genetic engineering technique.

[0020] Classified into the subphylum Fungi subphylum,
The microorganism capable of producing compound 1 can be cultured in a medium containing nutrients such as a carbon source and a nitrogen source that can be used by ordinary microorganisms.

As a carbon source, rice, glucose, water syrup, dextrin, sucrose, starch, molasses, animal and vegetable oils and the like can be used.

The nitrogen sources include soy flour, wheat germ,
Corn steep liquor, cottonseed meal, meat extract, peptone, yeast extract, ammonium sulfate, sodium nitrate, urea and the like can be used.

In addition, if necessary, it may be effective to add an inorganic salt capable of producing sodium, potassium, calcium, magnesium, cobalt, chlorine, phosphoric acid, nitric acid and other ions to the medium. .
In addition, an appropriate organic or inorganic substance that promotes the growth of bacteria and promotes the production of Compound 1 can be appropriately added to the medium.

The most suitable culture method for the microorganisms is a culture method under aerobic conditions, particularly a solid surface culture method using an agar medium or a rice medium, and a submerged culture method using a liquid medium. . The temperature suitable for culturing is 15 to 30 ° C, but preferably culturing at around 20 to 27 ° C. The production of compound 1 varies depending on the culture medium and culture conditions, but the accumulation in a liquid submerged culture in a flask usually reaches the maximum in 3 to 10 days. It is desirable to stop the culture when the amount of compound 1 accumulated in the culture reaches the maximum, and to separate and purify compound 1 from the culture.

The method for separating and purifying Compound 1 from the culture is not particularly limited, and any means available to those skilled in the art can be used. However, since Compound 1 is fat-soluble, By utilizing the properties, compound 1 can be efficiently separated from the culture and purified.

For example, a solvent extraction method using ethyl acetate, chloroform, etc .; synthetic adsorbents such as silica gel, alumina, ODS, Diaion HP-20 (manufactured by Mitsubishi Chemical);
Column chromatography using a gel filtration agent such as Sephadex LH-20 (manufactured by Pharmacia); high-performance liquid chromatography; and one or a combination of two or more such as preparative thin-layer chromatography using silica gel or the like as a carrier. It is effective to perform separation and purification.

Epicoccum purpurascen used in the method for producing compound 1 of the present invention
s) The D8646 strain (hereinafter, may be abbreviated as "D8646 strain") is a strain belonging to the order of the Imperial Mycoplasma subphylum Mycorrhizal fungi isolated from natural soil. Epicoccum purpurasce
ns) D8646 strain was deposited on May 16, 2000 with the Institute of Biotechnology, Industrial Science and Technology (Postal Code: 305-8566, 1-3-1 Higashi, Tsukuba, Ibaraki, Japan).
An accession number of MP-17859 has been assigned.

The details of the Epicoccum purpurascens strain D8646 will be described below. (1) Morphological properties of D8646 Characteristics of culture on various media i) Potato dextrose agar medium (PDA), 24
After culturing at 8 ° C. for 8 days, the colonies reach a diameter of 24 mm. The shape of the colony is velvety to fluffy, the color tone is yellow, and the portion where the conidial locus later formed has a dark olive color. The back of the colony is reddish-brown, producing a soluble dye in the medium, and the agar first turns lemon-colored and turns brown. ii) When cultured on Miura agar medium (LCA) at 24 ° C. for 8 days, colonies reach a diameter of 33 mm. The shape of the colony is fluffy and spreads thinly, the color is light yellow, and conidia are scattered. The back of the colony is yellow, and no production of soluble pigment is observed.

The conidium is black to dark brown and scatters or clumps on the medium. The conidiophore is formed on the conidial locus and is extremely short, club-shaped, smooth, 4.7-
5.9 μm.

The conidium is aleuro-type, brown to dark olive, and forms irregular septum in the vertical and horizontal directions, 7 to 15 cells, spherical, subspherical or pear type. Separation marks are seen on the surface, surrounded by warts, and the base is surrounded by relatively light-colored ring-shaped basal cells. The size is 5.9-10.9 × 5.6-10.
0 μm.

No teleomorph is observed. (2) Physiological properties Growth temperature range: 8-30 ° C, optimal growth temperature: 20 ° C (cultured on Miura agar medium for 4 days) Growth pH range: 3-10, optimal growth pH: 6 (cultured on Miura liquid medium for 4 days) (3) Taxonomic considerations This strain (D8646) forms multicellular conidia with vertical and horizontal partitions; conidia formation mode is aleuro-type; conidia peduncle hardly develops, conidia Clustering on the top; HL Ba based on the above features
'Illustrated genera of i' by rnett and BB Hunter
mperfect fungi '(1972) As a result of searching for genera using the search table on page 35, this strain was found to be epicoccam (Epoccam).
icoccum). Epicock (Epi
coccum) is epicoccum
cum purpurascens) is known to be the most widely distributed species. So, Shunichi Udagawa, Keisuke Tsubaki, and others,
When the morphological characteristics were compared with the description of this species on page 942 of the fungal illustration book (lower) (1978), the two strains were almost identical, and this strain (D8646) was identified as Epicoccum purpurascens. .

The compound 1 of the present invention has a telomerase inhibitory action and is used as an active ingredient of a drug based on inhibiting telomerase activity, preferably a drug for antitumor, more preferably a drug for treating malignant tumor. Useful. The telomerase inhibitory effect was determined by the method of Tatematsu et al. [Tatematsu, T. et al., Oncogene, 13, p2265 (1996).
Year), Japanese Patent Publication No. 8-018803] and the methods specifically described in the examples of the present specification.

When the medicament of the present invention is used as an antitumor medicament, the type of tumor to be treated is not particularly limited, and it can be applied to both solid tumors and non-solid tumors. The type of solid tumor or non-solid tumor is not particularly limited. For example, gastric cancer, esophageal cancer, duodenal cancer, small intestine cancer, large intestine cancer, liver cancer, gallbladder cancer, pancreatic cancer, kidney cancer, bladder cancer, oral cancer, bone cancer,
Examples include solid tumors such as brain tumor, skin cancer, breast cancer, uterine cancer, prostate cancer, and neuroblastoma, and non-solid tumors such as leukemia and malignant lymphoma. It is not limited.

As an active ingredient of the medicament of the present invention, a substance selected from the group consisting of compound 1, a physiologically acceptable salt of compound 1, and hydrates and solvates thereof can be used. it can.

As the physiologically acceptable salt of Compound 1, base addition salts can be used, and for example, metal addition salts such as sodium salt, potassium salt, magnesium salt and the like can be used.

Compound 1 and its salts may form hydrates, and may form solvates with methanol, ethanol, isopropanol, acetone, ethyl acetate, methylene chloride and the like. Are also included in the scope of the present invention.

As the active ingredient of the medicament of the present invention, a substance selected from the group consisting of compound 1, a physiologically acceptable salt of compound 1, and hydrates and solvates thereof is used as the medicament as it is. May be used, but usually
It is desirable to produce a pharmaceutical composition using one or more pharmaceutical additives.

The composition used in the medicament of the present invention may contain another telomerase inhibitor or an active ingredient of an antitumor medicament.

The administration route of the medicament of the present invention is not particularly limited, and either oral administration or parenteral administration can be selected. Pharmaceutical compositions suitable for oral administration include, for example, pharmaceutical forms such as tablets, capsules, granules, fine granules, powders, solutions, suspensions and the like. Pharmaceutical compositions suitable for parenteral administration include, for example, injections, drops, suppositories, inhalants, eye drops, ear drops, ointments, creams, transdermal absorbers, transmucosal absorbers, etc. Can be mentioned. However, the form of the medicament of the present invention is not limited to these.

[0040] The pharmaceutical composition can be formulated into a pharmaceutical composition according to a conventional method by using appropriate additives for the pharmaceutical composition according to the form.

For example, when preparing tablets which are solid preparations for oral administration, conventional excipients, binders, lubricants, coloring agents, disintegrators, etc., are used as additives for the preparation. Can be. As an excipient, for example, lactose, starch, talc, magnesium stearate, crystalline cellulose, methylcellulose, carboxymethylcellulose,
Glycerin, sodium alginate, gum arabic and the like can be mentioned, and as the binder, polyvinyl alcohol, polyvinyl ether, ethyl cellulose, gum arabic, shellac, sucrose and the like can be mentioned. Examples of the lubricant include magnesium stearate, talc and the like, and a coloring agent and a disintegrant can be appropriately selected from those usually used. Further, the tablet may be provided with an enteric coating or the like by a well-known method.

Solutions or suspensions, that is, liquid preparations such as aqueous or oily suspensions, solutions, syrups and elixirs can be prepared using a suitable aqueous medium by a commonly used method.

When preparing an injection for parenteral administration, a pharmaceutical additive such as a pH adjuster, a buffer, a stabilizer, an isotonic agent, a local anesthetic or the like is added, and the mixture is prepared in a conventional manner. It may be prepared as an injection for subcutaneous, intramuscular, or intravenous use.

When a suppository is produced, an oily base such as cacao butter, polyethylene glycol, lanolin, fatty acid triglyceride, witepsol (registered trademark of Dynamite Nobel) can be used as a base.

[0045] Capsules, granules, fine granules, powders, drops, inhalants, eye drops, ear drops, ointments, creams, transdermal absorbents, transmucosal absorbents, etc. are also commonly used. It can be prepared by a method.

The dose of the medicament of the present invention is not particularly limited, but the type of tumor to be treated, the condition of the patient, weight, age,
It is desirable to select appropriately depending on conditions such as whether or not another antitumor drug is used. For example, the amount of Compound 1 is preferably preferably in the range of about 1 to 2000 mg per adult day. The above dose can be administered in 1 to 4 times a day, and may be administered intermittently once every several days or several weeks.

[0047]

EXAMPLES Hereinafter, the present invention will be described more specifically with reference to Examples, but the scope of the present invention is not limited to the following Examples.

[0048]

Example 1 <Production of Compound 1> (1) Culture of D8646 Glucose 2.0%, glycerol 1.5%, corn starch 1.0%, dried yeast extract 0.35%, polypeptone 0.25%, soybean Powder 1.0%, sodium chloride 0.
3%, calcium carbonate 0.5% and traces of zinc sulfate,
Medium containing copper sulfate, manganese sulfate, antifoam (pH
6.0) was dispensed into 20 500 ml Erlenmeyer flasks at 80 ml each, and sterilized by high pressure steam at 121 ° C. for 20 minutes. The D8646 strain was inoculated into the culture, and cultured at 27 ° C. for 7 days with shaking at 210 rpm.

The D8646 strain was obtained on May 16, 2000.
The National Institute of Advanced Industrial Science and Technology (AIST) (Postal code 305-
8566 Deposited at 1-3-1, Higashi 1-chome, Tsukuba, Ibaraki, Japan, under the accession number of FERM P-17859. (2) Purification of Compound 1 Produced by Culture A 50% aqueous acetone solution was added to the Erlenmeyer flask that had been cultured, stirred, and allowed to stand at 4 ° C. overnight. 2.0 L of the obtained acetone extract was concentrated to 1.2 L under reduced pressure. The concentrated solution was extracted with 1.2 L of ethyl acetate, and the ethyl acetate layer was distilled off under reduced pressure to remove ethyl acetate.
17 g of powder were obtained.

The obtained acetone extract (1.23 L) was concentrated under reduced pressure to 1.2 L. This concentrated solution was adjusted to pH 4 with 1N hydrochloric acid, and extracted with 1.2 L of ethyl acetate. Ethyl acetate was distilled off from the ethyl acetate layer under reduced pressure to obtain 1.07 g of a red substance.

This material was dissolved in methanol and 2 g of O
DS (Wakogel 50C18: manufactured by Wako Pure Chemical Industries) was added, and the mixture was concentrated to dryness under reduced pressure. This is suspended in 20 ml of water, and 18 g
ODS was applied to a column packed with acetonitrile and washed well with water. After washing with 80 ml of water,
The mixture was washed with 100 ml of an acetonitrile mixture (4: 1) and 100 ml of a water-acetonitrile mixture (3: 2). Next, elution was performed with 100 ml of a water-acetonitrile mixture (2: 3).
The solvent was distilled off from this eluate under reduced pressure to obtain 0.30 g of a red substance.

[0052] This substance was used as Capcellpak C18 UG80 (30
mm: 250 mm) (manufactured by Shiseido). The red substance was dissolved in 5 ml of methanol, of which 1 ml was subjected to high performance liquid chromatography equilibrated with a water-methanol mixture (1: 1). Elution was carried out at 18 ml / min while changing the composition of the water-methanol mixture linearly from 1: 1 to 1: 9 in 30 minutes.
Fractions were fractionated. The solvent containing the fraction containing Compound 1 (fractions 65-66) was distilled off under reduced pressure to give Compound 1
Was obtained in an amount of 19.1 mg.

The physical properties of compound 1 thus obtained are as follows. 1) Appearance: red amorphous 2) Molecular weight: 612 3) Molecular formula: C ₃₄ H ₄₄ O ₁₀ 4) Mass spectrum: ESI 635 (M + Na) ⁺ , 651 (M + K) ⁺ 5) Hydrogen nuclear magnetic resonance spectrum: in deuterated methanol δppm: 7.13 (1H, dd, J = 14.5, 11.5Hz), 6.62 (1H, d
d, J = 14.0, 11.5Hz), 6.3-6.5 (7H, m), 6.20 (1H, d, J
= 15.0Hz), 6.13 (1H, s), 6.07 (1H, s), 5.60 (1H, d,
J = 10.5Hz), 4.54 (1H, d, J = 10.0Hz), 4.20 (1H, t, J
= 9.5Hz), 3.92 (1H, d, J = 2.5Hz), 3.7-3.8 (2H, m),
3.62 (1H, t, J = 6.0Hz), 3.53 (1H, dd, J = 9.5,3.0Hz),
3.0-3.1 (1H, m), 1.88 (3H, s), 1.1-1.4 (5H, m),
1.02 (3H, d, J = 6.5 Hz), 0.8-0.9 (6H, m) 6) Carbon nuclear magnetic resonance spectrum: in heavy methanol δppm: 172.01, 170.38, 166.10, 160.58, 149.02, 14
0.51, 140.30, 138.04, 137.50, 136.97, 136.64, 134.
03, 133.58, 132.30, 132.25, 131.37, 130.07, 122.5
0, 102.37, 101.81, 80.94, 76.55, 76.41, 71.00, 70.
42, 62.74, 45.81, 33.79, 33.09, 31.16, 21.74, 19.5
1, 13.78, 11.64

[0054]

Example 2 <Measurement of Telomerase Inhibitory Activity> Telomerase inhibitory activity of compound 1 obtained in Example 1 was measured. This measurement was performed according to Tatematsu K.
[Oncogene, 13, p2265 (1996), Tokuho 8-0]
18803] was modified as follows. (1) Preparation of telomerase extract A telomerase activity-positive tumor cell line PA-1 (American Ty) cultured in Dulbecco's modified minimum essential medium containing 10% inactivated fetal bovine serum at 37 ° C with 5% CO ₂ .
Pe Culture Collection (deposit number: CRL1572) was washed with phosphate buffered saline, and then separated from the culture dish and collected by centrifugation.

A cell lysate [10 mM Tris-HCl (pH 7.5), 1 mM MgCl ₂ , 0.5% 3-
[(3-cholamidopropyl) dimethylammonio] -1-propanesulfonate (3-[(3-cholamidepropyl) -dimethyl-
ammonio] -1-propanesulfonate (CHAPS)), 1m
M ethylene glycol bis (2-aminoethyl ether)
Tetraacetic acid (EGTA), 5 mM 2-mercaptoethanol, 10% glycerol, 0.1 mM 4- (2-aminoethyl) -benzenesulfonyl hydrochlorin (4- (2-amido)
noethyl) -benzenesulfonyl hydrochlorine (AEBS
F))) and suspended. After standing on ice for 30 minutes,
Centrifuged at 16,000 × g, 4 ° C. for 30 minutes. The supernatant was rapidly frozen in a dry ice / ethanol bath as a telomerase extract, and stored at -80 ° C until use. (2) Telomerase reaction Compound 1 (concentration range of 0.1 to 1000 μM) and 2 μm
M bpTG3 (substrate primer: biotinylated p
TG3 (5'-GTAAAACGACGGCCAGTTTGGGGTTGGGGTTGGGGTT
G-3 ') (SEQ ID NO: 1)) at a double concentration of a telomerase reaction solution (2 mM dTTP, 2 mM dATP, 2 m
M dGTP, 100 mM Trisacetic acid (pH 8.
0), 100 mM potassium acetate, 10 mM 2-mercaptoethanol, 2 mM MgCl ₂ , 2 mM EG
TA, 2 mM spermine, 2 mM spermidine) were mixed with an equal volume of a telomerase extract diluted with a cell lysate, and the mixture was incubated at 30 ° C. After a certain time, a final concentration of 16 mM
Then, ethylenediaminetetraacetic acid (EDTA) was added thereto and heated at 95 ° C. to stop the reaction (the obtained liquid was referred to as “telomerase reaction product”). Before starting the reaction, 20 mM EDTA was added in advance, and the value obtained under the condition that telomerase was inactivated was subtracted from the result as a background.

Further, dimethyl sulfo was used as a control.
Oxide was used in place of compound 1. (3) Preparation of avidin-coated PCR plate
Putavidin was added at 9 μl / well and 0.1 M 2-
[Morpholino] ethanesulfonic acid (2- [N-Mor
pholino] ethanesulfonic acid (MES) buffer (p
H6.0) dissolved in 1-ethyl-3- (3-dimethylaminopropane)
Ropyl) carbodiimide (1-ethyl-3- (3-dimethylaminop
ropyl) carbodiimide (EDC, Sigma))
Then, the mixture was kept at 37 ° C. for a certain time. Twin 2
Wash and dry with TBS containing 0 (Sigma) and sterile water
"Avidin cross-link plate". (4) Stretch PCR 25 μl of telomerase reaction product was
Telomerase reaction after incubation at 37 ℃
The product was bound to the plate and washed with distilled water. 25μ
1 / well of the PCR reaction solution [20 mM Tris-HCl (p
H8.3), 75 mM KCl, 0.005% W-1
(Surfactant), 1.5 mM MgCl _Two, 4 μM p
TG3 (sense primer), 1 μM pTAGγ (A
Antisense primer: 5'-CAGGAAACAGCTATGACCCCTAA
CCCTAACCCTAACCCT-3 ′), 50 μM dATP, 50
μM dGTP, 50 μM dCTP, 50 μM dT
TP, 1 U / 25 μl Taq DNA polymerase]
And use PCR Thermal Cycler MP (Takara Shuzo)
And performed PCR (93 ° C. for 1 minute, 69 ° C. for 1 minute, 72 ° C.)
30 cycles of 2 minutes). After completion of PCR, 25 μl of reaction solution
100 μl of sterilized water was added to the mixture to make a total of 125 μl (“PC
R product "). All primers and reagents used for PCR
Purchased from Life Tech Oriental. (5) Quantification of stretch PCR product 100 μl of PCR product was transferred to a 96-well black plate
To Bakelite) and dilute it 200 times with sterile water
100 µl of the prepared Pico Green (Molecular Probus)
After the mixture was left at room temperature for 5 minutes, the excitation wavelength was 485 nm and the absorption was
The fluorescence intensity at the collection wavelength of 538 nm
And fluoroscan II (Dainippon Pharmaceutical)
Was. As a result, 50% of compound 1 against telomerase
The inhibitory concentration was 67 μM, and Compound 1 was
It was found that it hindered the race.

[0057]

According to the present invention, it has been found that Compound 1 inhibits telomerase at a low concentration.

Accordingly, the medicament of the present invention has an action of suppressing the telomerase activity of tumor cells, and thus can be used as a telomerase inhibitor. Furthermore, it can be used as an antitumor drug, more preferably as an antineoplastic drug, for treating various tumors.

[0059]

[Sequence List] SEQUENCE LISTING <110> Mitsubishi-Tokyo Pharmaceuticals, Inc. (Mitsubishi Tokyo Pharmaceutical Co., Ltd.) <120> Telomerase inhibitor <130> YK00017 <160> 2 <210> 1 <211> 38 <212> nucleic acid <213> Artificial Sequence <220> <223> Sense primer <400> 1 gtaaaacgac ggccagtttg gggttggggt tggggttg 38 <210> 2 <211> 39 <212> nucleic acid <213> Artificial Sequence <220> <223> Antisense primer <400 > 2 caggaaacag ctatgacccc taaccctaac cctaacct 39

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) C12P 17/06 C12P 17/06 // C12N 1/14 C12N 1/14 A 15/09 ZNA (C12N 9 / 99 (C12N 9/99 C12R 1: 645) C12R 1: 645) (C12P 17/06 (C12P 17/06 C12R 1: 645) C12R 1: 645) (C12N 1/14 (C12N 1/14 C12R 1: 645) ) C12R 1: 645) C12N 15/00 ZNAA (72) Inventor Mika Kanda 1000 Kamoshidacho, Aoba-ku, Aoba-ku, Yokohama-shi, Kanagawa Prefecture Inside the Mitsubishi Chemical Research Laboratory (72) Inventor Masato Sugiyama Kamoshita, Aoba-ku, Yokohama-shi, Kanagawa 1000-cho, Mitsubishi Chemical Corporation Yokohama Research Laboratory F-term (reference) 4B024 AA11 CA01 HA14 4B064 AE46 AG21 BA07 BG04 BH04 BH05 CA05 CE08 CE10 DA05 4B065 A A57X AC14 BA22 CA18 CA44 4C062 DD22 4C086 AA01 AA02 AA03 BA17 GA02 MA01 MA04 NA14 ZB26 ZC20

Claims (6)

[Claims] A compound 1 represented by the following general formula (I):
A physiologically acceptable salt, hydrate, or solvate thereof. Embedded image 2. A telomerase inhibitor comprising the compound 1 according to claim 1, a physiologically acceptable salt thereof, a hydrate thereof or a solvate thereof as an active ingredient. 3. A medicament comprising the compound 1 according to claim 1, a physiologically acceptable salt thereof, a hydrate thereof or a solvate thereof as an active ingredient. 4. An antitumor medicament comprising the compound 1 according to claim 1, a physiologically acceptable salt thereof, a hydrate thereof or a solvate thereof as an active ingredient. 5. A compound 1 which is obtained by culturing a microorganism capable of producing the compound 1 according to claim 1, which belongs to the genus Epicoccum, and collecting the compound 1 from the obtained culture. Manufacturing method. 6. The microorganism capable of producing Compound 1 may be Epicoccum purpurasense (Epicoccum purpense).
urascens) strain D8646 (FERM P-1785)
9. The method according to claim 5, wherein 9).