KR101189756B1 - New Trichostatin analog and anti-tumor agent including this - Google Patents

Abstract

The present invention relates to a novel trichostatin derivative, and an antibiotic comprising the same, and more particularly, to a trichostatin derivative represented by Formula 1 or a pharmaceutically acceptable salt thereof, a method for preparing the same, and an effective thereof. The present invention relates to an antibiotic composition comprising the component. The novel trichostatin derivatives prepared using the bio- biotransformation technique using Streptomyces venezuelae from the trichostatin A of the present invention exhibit excellent antifungal and anticancer activity, and thus are useful as antibiotics or anticancer agents. Can be used.
[Formula 1]

Description

New Trichostatin Derivatives and Anticancer Agents Comprising the Same

The present invention relates to a novel compound and its use, and more particularly to a novel trichostatin derivative or a pharmaceutically acceptable salt thereof, a preparation method thereof, and an anticancer composition comprising the same as an active ingredient.

Trichostatin A is isolated from the soil bacterium Streptomyces hygroscopicus and is one of natural hydroxamates (see Non-Patent Document 1). Trichostatin A was originally found as an antifungal antibiotic and is also known as one of histone deacetylase inhibitors with broad spectrum epigenetic activity, and potential use as an anticancer agent is being considered (Non Patent Literature 2-4). Reference).

Indeed, over-activation of histone deacetylases in various cancer cell lines results in histone hypoacetylation, whereby histone deacetylase inhibitors specifically target histone deacetylases to prevent them. Make it functionally inactive. Several histone deacetylase inhibitors have been clinical trials for the treatment of leukemia and solid cancer, some of which have been approved and marketed. For example, there is a vorinostat, which is used for the treatment of T cell leaf species of the skin as a chemical synthetic mimic of trichostatin A (see Non-Patent Document 5-6).

Trichostatin A has previously been reported to exert anticancer activity against various cancer cell lines such as glioma cells and bladder cancer cells (see Non-Patent Document 7-9). Recently, it has been reported that Trichostatin A can be treated by reducing cell growth and proliferation of small cell lung cancer (SCLC) cell lines (see Non-Patent Document 10). Therefore, the potential of trichostatin A in the treatment of cancer raises interest in structurally modified trichostatin derivatives.

To achieve this, there can be two approaches to strategy; One is the chemical transformation approach of natural products, commonly referred to as the semi-synthetic method, and the other is the biotransformation approach using microorganisms as biocatalysts. Whereas chemical methods have low selectivity, low energy efficiency and environmentally unfriendly disadvantages exemplified by hydrogenation of rhodium metals of alkenes (Wilkinson's catalyst) (see Non-Patent Document 11), microbial conversion provides high selectivity, It has the advantage of being energy efficient and environmentally friendly.

However, there have been no reports of attempts to convert trichostatin A to microorganisms.

Recently, we have reported, by Streptomyces venezuelae , a unique, site-specific hydrogenation activity for unsaturated cyclic macrolides, which is a bacterial bio-hydrogenation system (bio-). The hydrogenation system can recognize specific structural embellishments around the target double bond, ie the carbonyl functionality of one adjacent carbon group and the methyl group of another adjacent carbon. (See the part indicated by the shaded rectangle of Formula 1). (See Non-Patent Document 12).

Therefore, the present inventors have studied to try to recognize and expand the unique biocatalyst practicality for trichostatin A, and as a result, Streptomyces S. venezuelae culture medium cultured in a medium containing trichostatin A From 2,3-dihydrotrichostatin A, which was previously unspecified, was isolated, the structure of the 2,3-dihydrotrichostatin A was analyzed, and the antifungal and antitumor activity The present invention was completed by confirming the same bioactive potential.

Tsuji, N; Kobayashi, M .; Nagashima, K., Wakisaka, Y .; Koizumi, K. J. Antibiot. 1976, 29, 1-6. Finnin, M. S .; Donigian, J. R .; Cohen, A .; Richon, V. M .; Rifkind, R. A .; Marks, P. A .; Breslow, R .; Pavletich, N. P. Nature 1999, 401, 188-193. Marks, P. A .; Richon, V. M .; Breslow, R .; Rifkind, R. A. Curr. Opin. Oncol. 2001, 13, 477-483. Drummond, D. C .; Noble, C. O .; Kirpotin, D. B .; Guo, Z .; Scott, G. K .; Benz, C. C. Annu. Rev. Pharmacol. Toxicol. 2005, 45, 495-528. Marks, P. A .; Breslow, R. Nat. Biotechnol. 2007, 25, 84-90. Codd, R .; Braich, N .; Liu, J .; Soe, C. Z .; Pakchung, A. A. Int. J. Biochem. Cell Biol. 2009, 41, 736-739. Kim, M. S .; Blake, M .; Baek, J. H .; Kohlhagen, G .; Pommier, Y .; Carrier, F. Cancer Res. 2003, 63, 7291-7300. Wetzel, M .; Premkumar, D. R .; Arnold, B .; Pollack, I. F. J. Neurosurg. 2005, 103, 549-556. Li, G. C .; Zhang, X .; Pan, T. J .; Chen, Z .; Ye, Z. Q. Int. J. Urol. 2006, 13, 581-586. Platta, C. S .; Greenblatt, D. Y .; Kunnimalaiyaan, M .; Chen, H. J. Surg. Res. 2007, 142, 219-226. Grushin, V. V. Acc. Chem. Res. 2010, 43, 160-171. Park, J. W .; Oh, H. S .; Jung, W. S .; Park, S. R .; Han, A. R .; Ban, Y. H .; Kim, E. J .; Kang, H. Y .; Yoon, Y. J. Chem. Commun. 2008, 44, 5782-5784. Ueki, M .; Teruya, T .; Nie, L .; Usami, R .; Yoshida, M .; Osada, H. J. Antibiot. 2001, 54, 1093-1095. Ommeslaeghe, K. V .; Elaut, G .; Breex, V .; Papeleu, P .; Iterbeke, K .; Geerlings, P .; Tourw, D .; Rogiers, V. Bioorg. Med. Chem. Lett. 2003, 13, 1861-1864. Oger, F .; Lecorgne, A .; Sala, E .; Nardese, V .; Demay, F .; Chevance, S .; Desravines, D. C .; Aleksandrova, N .; Le Gu, R .; Lorenzi, S .; Beccari, A. R .; Barath, P .; Hart, D. J .; Bondon, A .; Carettoni, D .; Simonneaux, G .; Salbert, G. J. Med. Chem. 2010, 53, 1937-1950. National Committee for Clinical Laboratory Standards (NCCLS). Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria That Grow Aerobically; Approved Standard M7-A5; NCCLS: Wayne, PA, 2000.

An object of the present invention is to provide a novel tricostatin derivative.

Another object of the present invention is to provide a method for preparing the tricostatin derivative.

Another object of the present invention to provide an antibiotic composition containing the trichostatin derivative or a pharmaceutically acceptable salt thereof as an active ingredient.

It is another object of the present invention to provide an anticancer agent composition containing the trichostatin derivative or a pharmaceutically acceptable salt thereof as an active ingredient.

In order to achieve the above object, the present invention provides a novel trichostatin derivative represented by the following formula (1) or a pharmaceutically acceptable salt thereof:

[Formula 1]

The present invention also provides a method for preparing the trichostatin derivative.

The present invention also provides an antibiotic composition containing the trichostatin derivative or a salt thereof as an active ingredient.

In addition, the present invention provides an anticancer composition containing the trichostatin derivative or a salt thereof as an active ingredient.

The present invention introduces a bioconversion technique of microorganisms, 2,3-dihydrotricostatin A, a novel tricostatin A derivative in which a double bond in a structure is substituted for trichostatin A, which is known to have existing antifungal and anticancer activity. The 2,3-dihydrotricostatin A exhibits antifungal activity similar to that of the conventional tricostatin A and exhibits twice the superior anticancer activity, thereby replacing the new 2,3-dihydrotricostatin A as an antibiotic or an anticancer agent. It can be seen that it can be usefully used as an effective ingredient of.

1 is a diagram showing the HPLC-ESI-MS chromatogram of the organic extract obtained from the culture medium in which S. venezuelae was cultured in the medium added to trichostatin A. Here, peaks labeled with trichostatin A and 2,3-dihydrotricostatin A were selected from molecules [M + H] + ions 303 and 305, respectively.
Figure 2 is a diagram showing the ESI-MS / MS spectrum of tricostatin A and its bioconversion derivative 2,3-dihydrotricostatin A.
FIG. 3 shows a ¹ H-NMR spectrum of trichostatin A and its bioconverting derivative 2,3-dihydrotricostatin A in CDCl ₃ :
a: trichostatin A; And
b: 2,3-dihydrotricostatin A.
FIG. 4 shows the ¹³ C-NMR spectrum of trichostatin A and its bioconverting derivative 2,3-dihydrotricostatin A in CDCl ₃ :
a: trichostatin A; And
b: 2,3-dihydrotricostatin A.
FIG. 5 shows Saccharomyces of Trichostatin A and its bioconversion derivative 2,3-dihydrotricostatin A. cerevisiae ) shows antifungal activity against yeast species.
Figure 6 is a diagram showing the anticancer activity of human SCLC DSM53 cell line of trichostatin A and its bioconversion derivative 2,3-dihydrotricostatin A.

EMBODIMENT OF THE INVENTION Hereinafter, this invention is demonstrated in detail.

The present invention provides a novel tricostatin derivative represented by the following formula (1).

The trichostatin derivative is preferably prepared by being bioconverted from Streptomyces venezuelae from Trichostatin A , but is not limited thereto, and includes all prepared by chemical synthesis.

Preferably, the trichostatin derivative has good antifungal activity against Saccharomyces sp. Yeast species, and more preferably, has excellent antifungal activity against Saccharomyces cerevisiae yeast species. Preferred but not limited to this.

The trichostatin derivative preferably has excellent anticancer activity against cancer cell lines, and more preferably has excellent anticancer activity against small cell lung cancer cell lines, but is not limited thereto.

The tricostatin derivative of the present invention represented by Chemical Formula 1 may be used in the form of a pharmaceutically acceptable salt, and as the salt, an acid addition salt formed by a pharmaceutically acceptable free acid is useful. Inorganic and organic acids can be used as the free acid, hydrochloric acid, bromic acid, sulfuric acid, phosphoric acid, etc. can be used as the inorganic acid, citric acid, acetic acid, lactic acid, maleic acid, fumaric acid, gluconic acid, methanesulfuric acid. Phonic acid, glyconic acid, succinic acid, tartaric acid, 4-toluenesulfonic acid, galluxuronic acid, embonic acid, glutamic acid or aspartic acid, and the like can be used. Preferably, hydrochloric acid is used as the inorganic acid, and methanesulfonic acid is used as the organic acid.

In addition, the tricostatin derivative represented by Formula 1 of the present invention includes not only pharmaceutically acceptable salts, but also all salts, hydrates, and solvates that can be prepared by conventional methods.

The addition salt according to the present invention can be prepared by a conventional method, for example, by dissolving the compound of Chemical Formula 1 in a water-miscible organic solvent such as acetone, methanol, ethanol, acetonitrile, etc., And then precipitating or crystallizing the acid solution. The solvent or excess acid may then be evaporated and dried in this mixture to obtain an addition salt or the precipitated salt may be prepared by suction filtration.

In addition, the present invention provides a method for producing a novel tricostatin derivative represented by the formula (1).

Specifically,

1) culturing the strain Streptomyces sp. In a medium to which trichostatin A is added;

2) extracting the culture solution cultured in step 1) with an organic solvent;

3) concentrating the organic extract of step 2) under vacuum; And

4) it can be prepared by a method comprising the step of dissolving the residue of step 3) in an organic solvent and then separating.

In the above method, the strain of Streptomyces sp. In step 1) is preferably Streptomyces venezuelae , but is not limited thereto.

In the above method, the organic solvent of step 2) is preferably ethyl acetate (EtOAc), but is not limited thereto.

In the above method, the organic solvent of step 4) is preferably methanol (MeOH), but is not limited thereto.

In the above method, the separation of step 4) may be performed using chromatography, and preferably, high-performance liquid chromatography (HPLC) is not limited thereto. In one embodiment of the present invention, the compound can be separated at a retention time of 35 to 38 minutes using preparative HPLC.

Production process according to the present invention is Streptomyces Venetian state elrae non-chemical synthesis (Streptomyces venezuelae ) It is characterized by manufacturing using bio-conversion technology using soil microorganisms.

In another aspect, the present invention provides an antibiotic composition containing a novel trichostatin derivative represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.

The antibiotic is preferably an antibiotic for fungi, and more preferably an antibiotic for fungi of Saccharomyces genus, but is not limited thereto.

In one embodiment of the present invention, the novel trichostatin derivatives or pharmaceutically acceptable salts thereof according to the present invention are similar to Saccharomyces cerevisiae in comparison to trichostatin A, which is known for its antibiotic activity. It showed antifungal activity. Therefore, it can be seen that the novel tricostatin derivative according to the present invention can be usefully used as a novel antibiotic.

In addition, the present invention provides an anticancer composition comprising a novel tricostatin derivative represented by the following Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

The cancer includes all leukemias or solid cancers overactive histone deacetylase, and is preferably any one selected from the group consisting of small cell lung cancer, breast cancer, colon cancer, pancreatic cancer, ovarian cancer, prostate cancer and leukemia. Most preferred is small cell lung cancer, but is not limited thereto.

In one embodiment of the present invention, the novel trichostatin derivatives or pharmaceutically acceptable salts thereof according to the present invention exhibited at least two times better anticancer activity against small cell lung cancer compared to trichostatin A, which is known for its existing anticancer activity. . Therefore, it can be seen that the novel tricostatin derivative according to the present invention can be usefully used as a new anticancer agent.

When the composition of the present invention is used as a medicine, the pharmaceutical composition containing the trichostatin derivative represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient may be used for various oral or parenteral administrations during clinical administration. It may be formulated in a form and administered, but is not limited thereto.

Formulations for oral administration include, for example, tablets, pills, hard / soft capsules, solutions, suspensions, emulsifiers, syrups, granules, elixirs, and the like. Rose, sucrose, mannitol, sorbitol, cellulose and / or glycine), lubricants such as silica, talc, stearic acid and its magnesium or calcium salts and / or polyethylene glycols. Tablets may also contain binders such as magnesium aluminum silicate, starch paste, gelatin, methylcellulose, sodium carboxymethylcellulose and / or polyvinylpyrrolidine and may optionally contain additives such as starch, agar, alginic acid or its sodium salt A disintegrating or boiling mixture and / or an absorbent, a colorant, a flavoring agent, and a sweetening agent.

The pharmaceutical composition comprising the trichostatin derivative represented by Formula 1 as an active ingredient may be administered parenterally, and the parenteral administration may be by injection of subcutaneous injection, intravenous injection, intramuscular injection, or intrathoracic injection.

At this time, to prepare a formulation for parenteral administration, the trichostatin derivative of Formula 1 or a pharmaceutically acceptable salt thereof is mixed with water with a stabilizer or buffer to prepare a solution or suspension, which is an ampoule or vial unit dosage form. It can be prepared by. The compositions may contain sterile and / or preservatives, stabilizers, hydrating or emulsifying accelerators, auxiliaries such as salts and / or buffers for the control of osmotic pressure, and other therapeutically useful substances, and conventional methods of mixing, granulating It may be formulated according to the formulation or coating method.

In addition, the dosage of the compound of the present invention to the human body may vary depending on the age, weight, sex, dosage form, health condition and degree of disease of the patient, and generally based on an adult patient having a weight of 70 kg. 0.001 to 1,000 mg / day, preferably 0.01 to 500 mg / day, and may be administered once a day to several times at regular intervals according to the judgment of a doctor or pharmacist.

Hereinafter, examples for explaining the present invention in more detail.

However, the following examples are merely provided to explain the present invention more easily, and the content of the present invention is not limited by the examples.

< Example  1> Tricostatin  Preparation of A Derivatives

<1-1> Tricostatin  Biotransformation of A

Recombinant mutant species of S. venezuelae YJ028 [Park, JW et al ., Chem . Commun . 2008, all genes encoding pikromycin polyketide synthase and desamine amino biosynthesis genes are removed. and 44, using the species described in the 5782-5784) a first, SPA (sorbitol pyroglutamic acid) agar plates [0.1% yeast extract, 0.1% beef extract, 0.2% lactose trip (tryptose), 1.0% glucose (glucose), 1.5% agar, and traces of FeSO ₄ ] were streaked for isolation and then incubated at 30 ° C. for 2 days. Then in a partitioned Erlenmeyer flask, 50 ml of SCM medium [1.5% soluble starch, 2.0% soytone, 0.01% CaCl ₂ , 0.15% yeast extract for 2 days at 30 ° C. (yeast extract, and 1.0% MOPS) was added tricostatin A (Sigma, St. Louis, MO) (250 μg; final concentration 5 μg / mL; dissolved in 100 uL MeOH) , Incubated for 3 more days. All experiments were performed at least three times independently.

<1-2> Tricostatin  Extraction and Separation of A Derivatives

The whole culture of Example <1-1> was extracted and separated twice with an average volume of EtOAc in a 250 mL separatory funnel, and then the organic extract was concentrated in vacuo. The dried residue was immediately dissolved in 200 uL MeOH and part of the solvent was used for analysis. First of all, Micromass Quattro micro Analysis was performed using a Waters / Micromass Quattro micro / MS connector consisting of an analytical reversed-phase Nova-Pak C ₁₈ column (Waters, Milford, MA; 150 × 3.9 mm, 4.0 μm) directly connected to MS.

As a result, it was confirmed that addition of the tricostatin A to various batch cultures of YJ028 species resulted in a reduced tricostatin derivative corresponding to tricostatin A at a conversion rate of about 18% (see FIG. 1).

These new trichostatin derivatives were then purified by preparative reversed phase HPLC, and chromatographic separation using successive ESI-MS analysis of each fraction collected (5 mL / fraction), the chemical structures of which were ¹ H- and ^{It was} analyzed using ¹³ C-NMR spectroscopy. Specifically, chromatographic separation was performed using preparative HPLC consisting of Watchers 120 ODS-BP column (250 × 10.0 mm, 5.0 μm). In addition, ¹ H- and ¹³ C-NMR spectroscopy were performed as follows. NMR samples were prepared by dissolving a trichostatin derivative in 200 uL of CDCl ₃ and then leaving the solvent in a 5 mm Shigemi advanced NMR microtube (Sigma, St. Louis, MO) appropriate for the solvent. ¹ H and ¹³ C NMR spectra were obtained at 298 K using a Varian INOVA 500 spectrophotometer, and chemical shifts were recorded in ppm using TMS as internal reference. All NMR data calculations were performed using Mnova Suite 5.3.2 software.

The MS / MS spectrum of Trichostatin A at m / z 303, representing proton molecular ions, showed two or four fragment ions at m / z 148, 177, and 270, exactly as previously reported (see Non-Patent Document 13). . On the other hand, the MS / MS spectrum of the trichostatin derivative shows some fragment ions at m / z 148 and 177, which is typical of trichostatin A, as well as a difference of 2 Da from the generated ions at m / z 270 generated from trichostatin A. I showed specific product ions at m / z 272 (see FIG. 2). This result suggests that one of the two double bonds present in C2, 3 and C4, 5 is reduced in the trichostatin derivative.

NMR spectral data of Trichostatin A and its derivatives 2,3-dihydrocostatin A (500 MHz, CDCl ₃ ) 2,3-dihydrotricortin A Trichostatin A location δ _C , mult δ _H , mult δ _C δ _H One 172.2, C - 166.1 - 2 029.8, CH ₂ 2.26, t 117.5 5.27 2.22, ddd 3 036.2, CH ₂ 2.20, m 145.0 7.41 2.17, ddd 4 134.1, C - 132.8 - 5 133.6, CH 5.47, d 140.1 5.68 6 040.9, CH 3.83, dq 40.8 3.84 7 199.6, C - 199.4 - 8 123.7, C - 123.8 - 9, 13 130.8, CH 7.78, d 130.8 7.78 10, 12 110.8, CH 6.82, d 110.8 6.83 11 153.6, C - 153.6 - 14 16.0, CH ₃ 1.82, s 12.6 2.21 15 17.7, CH ₃ 0.94, d 17.8 0.93 16 39.9, CH ₃ 3.05, s 39.9 3.05

Through comparison of the ¹ H-NMR spectral data of Trichostatin A and its reduced derivatives, the most obvious changes observed in these derivatives are typical for olefinic protons (H-2,3) in Trichostatin A. There was a lack of signal at 5.27 and 7.41 ppm. Upfield shifts of the C-2 and C-3 signals of the parent compound (29.8 and 36.2 ppm at 117.5 and 145.0 ppm, respectively) support the differences identified through the ¹ H-NMR data. This demonstrates that the bioconversion derivatives have reduced C-2,3 double bonds (see FIG. 3). This result suggests that 2,3-dihydrotrichostatin A has a site-selective activity for biohydrogenation for unsaturated linear chain-like hydroxamate as well as various ring macrolides. (regio-selective activity), including the conversion of S. venezuelae is to show that (see Non-Patent Document 12).

< Example  2> Tricostatin  Identification of antifungal activity of A derivative

To evaluate the antifungal activity of the tricot A statin derivatives, ATCC (American Type Culture Collection) (Manassas, VA) a saccharide in my process three rebiah claim (Saccharomyces obtained from cerevisiae ) Antifungal activity was confirmed using ATCC 9763.

Specifically, Saccharomyces ( Saccharomyces) cerevisiae ) ATCC 9763 was incubated and maintained in Antibiotic 19 (Difco, BD Biosciences, San Jose, CA) at 30 ° C. Tricostatin A and its corresponding 2,3-dihydrotricostatin A were then treated with Saccharomyces cerevisiae. Antimicrobial activity against Saccharomyces cereviase was measured using a microdilution method recommended by the Clinical and Laboratory Standard Institute (CLSI) (see Non-Patent Document 16). Continuous 2-fold dilutions of these hydroxamic acids were performed using DMSO to a final concentration of 12.5-200 μg / mL, and a partial sample of DMSO (final 0.1%) was used as a negative control. Saccharomyces cerevisiae growth was monitored at 600 nm using a Labsystems Bioscreen C reader, and the lowest concentration of hydroxamate diluted in broth medium inhibiting the growth of test microorganisms was determined using MIC. It was. All analyzes were performed three times.

MIC Comparison of Trichostatin A and Its Derivatives 2,3-Dihydrotricostatin A Trichostatin A 2,3-dihydrotricortin A MIC (/ mL) 50.0 50.0

As a result, 2,3-dihydrotricostatin A of the present invention showed excellent antimicrobial activity similar to tricostatin A. It can be seen that the deficiency of C2,3-double bond in trichostatin A does not affect the antifungal activity (see Table 2 and FIG. 5).

< Example  3> Tricostatin  Anticancer activity of A derivative

In order to determine the anticancer activity of the trichostatin A derivative, anticancer activity was confirmed using a human SCLC DSM53 cell line obtained from the American Type Culture Collection (ATCC) (Manassas, VA).

Specifically, the human SCLC DSM53 cell line is 10% fetal bovine serum (Sigma), penicillin (penicillin) and streptomycin (Invitrogen) added Waymouth's MB752 / 1 medium (Invitrogen, San Diego, CA) ) (See Non-Patent Document 10). The cell line was incubated in a humid atmosphere at 37 ° C., 5% CO ₂ . Then, after treatment with Trichostatin A and its corresponding 2,3-dihydrocostatin A, respectively, proliferation of DMS53 cells was confirmed by MTT [3,4- (4,5 dimethylthiazol-2-yl) -2,5 -diphenyltetrazolium bromide] analysis (see Non-Patent Document 10). In summary, DMS53 cells were trypsinized and then inoculated into 24-well plates and left overnight. Continuous 2-fold dilutions of these hydroxamic acids were performed using DMSO to a final concentration of 3.1-200 nM, and a partial sample of DMSO was used as a negative control. After 2 days of treatment, the medium was removed and then replaced with a medium containing MTT, followed by further incubation for 2 hours, followed by monitoring at absorbance at 540 nm. IC ₅₀ is defined as the lowest concentration of Trichostatin A and 2,3-dihydrotricortatin A when the growth of DMS53 cells is 50% inhibited compared to the control cell line grown in medium not treated with any hydroxamate. All analyzes were performed three times.

Comparison of IC ₅₀ of Trichostatin A and its Derivatives 2,3-DihydroCostatin A Trichostatin A 2,3-dihydrotricortin A IC ₅₀ (nM) 25.0 25.0

As a result, the tricot statin A, on the other hand represents the IC ₅₀ ~ 25.0 nM, 2,3- dihydro-tricot statin A exhibited IC ₅₀ ~ 12.5 nM. That is, 2,3-dihydrocostatin A showed a two-fold increase in anticancer activity against human SCLC DMS53 cells compared to tricostatin A. It can be seen that the deficiency of C2,3-double bonds in Trichostatin A increases anticancer activity (see Table 3 and Figure 6).

In conclusion, the present invention is the first to produce trichostatin A derivatives using bio-biotransformation techniques by microorganisms, instead of the total- or semi-synthesis methods commonly used to produce novel trichostatin derivatives and / or homologues. It is a report. Further, Streptomyces a Venetian state elrae (S. venezueale) site-specific biological activity curing the first report apply to the similar linear hydroxy roksa formate not limited to fluoride unsaturated ring mark of. Therefore, as one of the scaffold retrofit approaches, it can be seen that such Streptomyces Venezuelan bioconversion system may be helpful in the development of new and / or enhanced therapeutic resources.

On the other hand, 2,3-dihydrotricostatin A represented by the formula (1) according to the present invention can be formulated in various forms according to the purpose. The following are some examples of formulation methods containing the compound represented by Formula 1 according to the present invention as an active ingredient, but the present invention is not limited thereto.

< Formulation example  1> tablet (direct pressure)

After sifting 5.0 mg of the active ingredient, 14.1 mg of lactose, 0.8 mg of crospovidone USNF, and 0.1 mg of magnesium stearate were mixed and pressed to prepare a tablet.

< Formulation example  2> tablets (wet assembly)

After 5.0 mg of the active ingredient was sieved, 16.0 mg of lactose and 4.0 mg of starch were mixed. 0.3 mg of Polysorbate 80 was dissolved in pure water, and an appropriate amount of this solution was added, followed by atomization. After drying, the granules were sieved and mixed with 2.7 mg of colloidal silicon dioxide and 2.0 mg of magnesium stearate. The fine particles were pressed to prepare tablets.

< Formulation example  3> with powder Capsule

5.0 mg of the active ingredient was sieved and then mixed with 14.8 mg of lactose, 10.0 mg of polyvinylpyrrolidone and 0.2 mg of magnesium stearate. The mixture was extruded through a hard No. Filled in 5 gelatin capsules.

< Formulation example  4> Injection

Injectables were prepared by containing 100 mg as active ingredient, as well as 180 mg of mannitol, 26 mg of Na ₂ HPO ₄ 12H ₂ O and 2974 mg of distilled water.

The present invention can be usefully used in the development of a variety of anticancer agents or antibiotics by showing the expandability of the microbial hydrogenation activity, it can be useful in the development of bioconversion technology as one of the scaffold modification strategy.

Claims (14)

Trichostatin derivatives represented by Formula 1 below:
[Formula 1]

The method of claim 1,
Trichostatin derivative, characterized in that the biotransformation produced by Streptomyces venezuelae from Trichostatin A.
The method of claim 1,
Trichostatin derivative, characterized by having antifungal activity against Saccharomyces cerevisiae .
The method of claim 1,
Trichostatin derivative, characterized in that it has anticancer activity against small cell lung cancer (SCLC).
1) culturing the strain Streptomyces sp. In a medium to which trichostatin A is added;
2) extracting the culture solution cultured in step 1) with an organic solvent;
3) concentrating the organic extract of step 2) under vacuum; And
4) A process for preparing the trichostatin derivative of claim 1, comprising the step of dissolving the residue of step 3) in an organic solvent and then separating.
6. The method of claim 5,
Streptomyces genus strain of step 1) is characterized in that the Streptomyces Venezuelan ( S. venezuelae ).
6. The method of claim 5,
The organic solvent of step 2) is characterized in that the ethyl acetate (EtOAc).
6. The method of claim 5,
The organic solvent of step 4) is characterized in that methanol (MeOH).
6. The method of claim 5,
Separation of step 4) is characterized in that using chromatography.
delete An antibiotic composition comprising the trichostatin derivative of claim 1 or a pharmaceutically acceptable salt thereof as an active ingredient.
12. The method of claim 11,
Said composition is an antibiotic composition, characterized in that it exhibits antifungal activity against Saccharomyces cerevisiae .
An anticancer agent composition comprising the trichostatin derivative of claim 1 or a pharmaceutically acceptable salt thereof as an active ingredient.
The anticancer composition according to claim 13, wherein the cancer is any one selected from the group consisting of small cell lung cancer, breast cancer, colon cancer, pancreatic cancer, ovarian cancer, prostate cancer and leukemia.

Images