KR100603024B1 - Temperature-sensitive triphosphazene-5-fluorouracil conjugate and preparation method thereof - Google Patents

Abstract

The present invention relates to a novel temperature sensitive cyclic phosphazene trimer-5-fluorouracil complex and a method for preparing the same, wherein the temperature sensitive cyclic phosphazene trimer-5-fluorouracil complex has an excellent anticancer effect. In addition, since it has a stereoselective structure and can control temperature sensitivity, not only systemic administration but also targeted local administration is possible.

Description

TEMPERATURE-SENSITIVE TRIPHOSPHAZENE-5-FLUOROURACIL CONJUGATE AND PREPARATION METHOD THEREOF}

The present invention relates to a novel temperature sensitive cyclic phosphazene trimer-5-fluorouracil complex and a method for preparing the same, wherein the temperature sensitive cyclic phosphazene trimer-5-fluorouracil complex has an excellent anticancer effect. In addition, since it has a stereoselective structure and can control temperature sensitivity, not only systemic administration but also targeted local administration is possible.

Temperature sensitivity refers to a reversible phenomenon in which a substance dissolves in water at a low temperature, and solubility rapidly decreases and precipitates when it is above a certain threshold temperature, and when the temperature is lowered again, it is dissolved in water. Is referred to as Low Critical Solution Temperature (LCST) or cloud point. Such temperature-sensitive characteristics are actively researched in various fields such as medical materials, thin films, separation from biochemical reactions, cosmetics, and optics.

Until now, most of the temperature-sensitive substances were organic hardly decomposable polymers, but several biodegradable polymers have been reported since several years [eg, Jeong, B. et al ., Nature , 388 , 860 (1997), Song, SC et al., Macromolecules , 32 , 2188 (1999), Lee, SB et al., Macromolecules , 32 , 7820 (1999)]. 5-Fluorouracil (5-Fu), first synthesized by Yasmoto et al., Has been known as an anticancer agent that produces interferon. The most commonly used anticancer agents for clinical use include doxorubicine, cisplatin and 5-fluorouracil.

Drug efficacy varies depending on the type of cancer being used in the clinic, and it is known that there are many differences in the mechanism of action and side effects. In particular, it is known that 5-Fu is highly toxic, accompanied by various side effects, and fast excretion into the body after administration, resulting in poor sustainability of the drug. Therefore, there is an urgent need for the development and synthesis of 5-Fu derivatives having low toxicity and long residence time, and excellent pharmacological effects.

An object of the present invention is to develop a novel third generation temperature sensitive anticancer agent which is excellent in anticancer effect and low in toxicity. To this end, the present inventors introduced a solubilizing agent and a functional group into the cyclic phosphazene trimer, and then introduced a 5-fluorouracil derivative into the cyclic phosphazene trimer having the functional group, thereby accurately reducing the low critical solution temperature. The present invention has been accomplished by finding that a temperature-sensitive cyclic phosphazene trimer-5-fluorouracil complex anticancer agent that can be variously controlled and can be targeted by systemic and local administration can be obtained.

The present invention relates to a novel temperature sensitive cyclic phosphazene trimer-5-fluorouracil complex and a method for preparing the same, wherein the temperature sensitive cyclic phosphazene trimer-5-fluorouracil complex has an excellent anticancer effect. In addition, since it has a stereoselective structure and can control temperature sensitivity, not only systemic administration but also targeted local administration is possible.

More specifically, the present invention has a stereoselective structure which is a non-geminal and cis-isomer, exhibits various temperature sensitivity in a range including the temperature of the body temperature region, and is represented by the following general formula (I) A cyclic phosphazene trimer-5-fluorouracil complex is provided, having the structure represented.

In the formula, m is an integer selected from 2 and 7 as the number of repeating units of the ethoxy group of the poly (alkoxyethylene glycol), n is an integer selected from 0 or 1, 2 and 3, indicating the length of the alkyl chain. R is an amino acid residue group such as H (glycine acid group), CH ₃ (alanine acid group), CH (CH ₃ ) ₂ (valine acid group), CH ₂ CH (CH ₃ ) ₂ (leucine acid group), and It is selected from CH ₂ C ₆ H ₆ (phenylalanine group) is preferred.

 5-Fluorouracil complexes exhibiting various temperature sensitivity of the present invention are novel substances with release control that have not yet been published, particularly local drug delivery using their temperature sensitivity as well as systemic administration. This has the advantage that targeting drug delivery is possible. Thus, it is expected that breakthrough new therapies will be possible in the treatment of cancer, in particular in the treatment of solid cancer.

In the cyclic phosphazene trimer-5-fluorouracil complex of the present invention, their low-critical solution temperature varies depending on the type of poly (alkoxyethylene glycol) and amino acids used and the like. And by using amino acids, the low-critical solution temperature of the cyclic phosphazene trimer-5-fluorouracil complex can be adjusted to suit the application.

In addition, the present invention has a stereoselective chemical structure by substituting the chlorine atom of hexacyclotriphosphazene with poly (alkoxyethylene glycol) and lysine ethyl ester, and then combining 5-fluorouracil derivatives having amino acids. The present invention provides a method for preparing a temperature sensitive cyclic phosphazene trimer-5-fluorouracil complex in oligomeric form, capable of precisely controlling a low critical solution temperature. Said amino acids are glycine, alanine, valine, leucine and Preferred is selected from phenylalanine and, as described above, by using the appropriate poly (alkoxyethyleneglycol) and amino acids, the temperature sensitive cyclic phosphazene trimer-5-fluorouracil whose low critical solution temperature is adapted to the application The preparation of the complex is possible.

More specifically, in the production method of the present invention, hexacyclotriphosphazene is reacted with hydrophilic poly (alkoxyethylene glycol) having different molecular weights, and then lysine ethyl ester is subjected to nucleophilic substitution reaction to form cyclic phosphazene having a functional group. Synthesizing the sieve, followed by nucleophilic substitution reaction of a 5-fluorouracil derivative having an amino acid bound thereto, to obtain a cyclic phosphazene trimer-5-fluorouracil complex exhibiting various temperature sensitivity. .

More specifically, the manufacturing method of the cyclic phosphazene trimer-5-fluorouracil complex which has the stereoselective chemical structure represented by General formula (I) of this invention,

Hexacyclotriphosphazene, a cyclic phosphazene trimer, is reacted with a hydrophilic poly (alkoxyethylene glycol), followed by nucleophilic substitution reaction of lysic acid ethyl ester to give a cyclic phosphazene trimer intermediate having the structure of formula (II). To prepare a compound;

A compound having the structure of formula (VI) by reacting a compound having the structure of formula (III), a compound having the structure of formula (IV), and a compound having the structure of formula (V) Manufactures:

Reacting the intermediate compound having the structure of the general formula (II) with the compound having the structure of the general formula (VI) to prepare a compound having the structure of the following general formula (VII);

Reducing the compound of formula (VII) obtained above using a palladium / charcoal catalyst under hydrogen gas to prepare a temperature sensitive cyclic phosphazene trimer-5-fluorouracil complex having the structure of formula (I). It includes.

In General Formulas (II) and (III), m is an integer selected from 2 and 7 as the number of repeating units of the ethoxy group of the poly (alkoxyethylene glycol), and n represents the length of the alkyl chain. And an integer selected from 3. R is an amino acid residue group such as H (glycine acid group), CH ₃ (alanine acid group), CH (CH ₃ ) ₂ (valine acid group), CH ₂ CH (CH ₃ ) ₂ (leucine acid group), and It is selected from CH ₂ C ₆ H ₆ (phenylalanine group) is preferred.

When the above manufacturing process is represented by the reaction scheme, the following process is the same as FIG.

The intermediate compound having the structure of Formula (II) may be prepared by reacting hexacyclotriphosphazene with a hydrophilic poly (alkoxyethylene glycol), and then nucleophilic substitution reaction of the obtained reaction product using an excess amount of lysine ethyl ester. Can be.

The reaction ratio of hexacyclotriphosphazene and poly (alkoxyethylene glycol) is preferably in a molar ratio of 1: 2.8 to 1: 3.2, more preferably 1: 3. This is because, in one unit of hexacyclotriphosphazene, there are a total of six reactive functional groups, and half of these three functional groups can induce a uniform reaction when reacted with polyalkoxyethylene glycol, The physical properties of the material are best maintained.

In this reaction, an excess of lysine ethyl ester is added to react the lysine ethyl ester with the functional group of hexacyclotriphosphazene remaining in the reaction product without reacting with polyalkoxyethylene glycol. The lysic acid ethyl ester to be added is preferably added in an amount of 2 to 5 times, preferably 4 times, the unreacted hexacyclotriphosphazene in consideration of three functional groups. Therefore, the amount of lysine ethyl ester added to the reaction product is 6 times the number of moles of the reaction product (two times the unreacted hexacyclotriphosphazene functional group) to 15 times (five times the unreacted hexacyclotriphosphazene functional group). Preferred is most preferably 12 times (4 times the unreacted hexacyclotriphosphazene functional group).

Optionally, in order to increase the purity of the compound having the structure of Formula (II), the obtained product may be dissolved in a small amount of chloroform, and then excess hexane is added to precipitate the final product to obtain a compound of Formula (II). . In order to further increase the purity of the compound of formula (II), the precipitated material is dissolved in distilled water to precipitate a substance (eg, CF ₃ CH ₂ OH, etc.) having a low critical temperature according to the low critical solution temperature of each product. This can be introduced in an amount that can be induced to precipitate, followed by centrifugation to obtain a compound of formula (II) having increased purity.

Apart from this, the compound having the structure of formula (III) may be prepared according to the method described, for example, in "Putnam, D .; Kopecek, J. Bioconjugate. Chem. 1995 , 6, 483".

In the preparation of the compound having the structure of formula (VI), the compound having the structure of formula (III), the compound having the structure of formula (IV) and the compound having the structure of formula (V) are 1: It is preferable to react in a molar ratio of 1: 1. This reaction is a step of synthesizing a precursor for reacting 5-fluorouracil with the amine group of lysine, and as described above, it is preferable to react each compound in the same amount, otherwise the reaction is uniform. It is undesirable because there is the possibility of side reactions that are not supported or desired.

In the preparation of the compound having the structure of formula (VII), the intermediate compound having the structure of formula (II) is 1: 3 to 1: 4, preferably the compound having the structure of formula (VI). Preferably, the reaction is carried out at a molar ratio of 1: 3.2 [Compound of Formula (II): Compound of Formula (VI)]. In the above reaction, hexacyclotriphosphazene is substituted with three lysic acid ethyl esters (Compound II), and in order to react the 5-fluorouracil precursor (Compound VI) at a ratio of 1: 1, It is appropriate to react the compound in a ratio of 1: 3, and in view of the degree of reaction thereof, it is preferable to further add a small amount of 5-fluorouracil precursor, and it is preferable to react with the reaction ratio in the above range. However, even if the 5-fluorouracil precursor is added in the above range, the amount to be reacted is constant and unnecessary loss occurs, so it is not preferable to add in the above range. In this way, the obtained product is dissolved in methanol, and unreacted substances and by-products are removed using a dialysis membrane (MW, cutoff = 2,000) to obtain a more pure compound of formula (VII).

In the step of reducing the compound of formula (VII) to prepare a compound of formula (I), the obtained product is dissolved in methanol, and then used for 4 to 5 hours under hydrogen gas using a 10% palladium / charcoal catalyst. It is preferable to make it react. After dissolving the obtained product in water, only the filtrate (dissolved portion) can be lyophilized to obtain the final product. This reaction is a step of reducing the material before obtaining the final product, in which all organic solvents can be used, and methanol is most preferably used as a solvent in order to induce the desired reduction reaction in the most stable state.

In more detail, the steps of 3.2 moles of N, N'-dicyclohexylcarbodiimide having 3.2 moles of the compound of general formula (III) and the following general formula (IV) and the following general formula (V) After 3.2 mol of N-hydroxysuccinimide having a structure is reacted in a tetrahydrofuran solvent for 24 hours, the reaction product is added dropwise to 1 mol of a compound solution of the general formula (II) dissolved in the same solvent. After the mixture was reacted for 24 hours, the solvent was removed by distillation under reduced pressure, and the obtained product was dissolved in methanol, and then unreacted material was removed using a dialysis membrane (MW, cutoff = 2,000). After dialysis for 48 hours, the concentrated solution by distillation under reduced pressure is stirred for 4-5 hours with palladium / charcoal catalyst under hydrogen gas. After distillation under reduced pressure to remove the solvent, it is dissolved in water. The solution obtained by filtering only the part dissolved in water is lyophilized to obtain a final cyclic phosphazene trimer-5-fluorouracil complex having a structure of general formula (I).

The present invention also provides compositions for cell proliferation inhibitors containing an effective amount of the cyclic phosphazene trimer-5-fluorouracil complex as the active ingredient. The composition of the present invention has an excellent cell proliferation inhibitory effect, particularly against tumor cells (see Table 1). Because the cyclic phosphazene trimer-5-fluorouracil complex is stereoselective and temperature sensitive, the composition of the present invention containing it is capable of targeted local administration as well as systemic administration.

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

In the following examples, the carbon, hydrogen and nitrogen elemental analysis of the compound of the present invention was performed by EA1110 (CE Instrument, italy) Perkin-Elmer C, H, N analyzer of Seoul National University instrument source, HPLC was KIST It was performed by a 3.9 mm × 300 mm C18 column (developing solvent: 100% methanol) in the characterization center. Hydrogen and phosphorus nuclear magnetic resonance spectra were measured using a Varian Gemini-300, and the low critical solution temperature was measured using a Perkin-Elmer Lamda18 UV / VIS spectrometer.

Example 1

[(Poly (methoxyethyleneglycol350)] [L-Louisyl-2- (5-fluorouracil-1-yl) -L, D-glycyl-L-lysineethylester] cyclotriphosphazene, { NP [O (CH ₂ CH ₂ O) ₇ CH ₃ ] [Lys-Gly (5-Fu) -Leu]} ₃ Manufacture

Poly (methoxyethyleneglycol) ethanol (1.8 g, 5.0 mmol) and sodium metal flakes (0.14 g, 6.0 mmol) having a molecular weight of 350 were added to 150 mL of dried tetrahydrofuran and refluxed under a stream of nitrogen for 24 hours. The sodium salt of (methoxyethylene glycol) ethoxide was prepared. Hexacyclotriphosphazene (0.50 g, 1.4 mmol) was dissolved in 10 mL of dried tetrahydrofuran, and then placed in a dry ice-acetone bath and half of the poly (methoxyethylene glycol) ethoxide sodium salt solution prepared above. Was added drop wise.

After reacting for 6 hours at room temperature, 1 mL of the solution was taken to obtain ³¹ P-NMR spectra. The poly (methoxyethylene glycol) ethoxide sodium salt solution prepared above was added dropwise until one peak appeared. When the above procedure is repeated and the peaks become one, the solution is dissolved in 150 mL of tetrahydrofuran dried with lysine ethyl ester (3.0 g, 17 mmol) and triethylamine (1.2 mL, 8.4 mmol). Dropped in

After the reaction was stirred at room temperature for 48 hours, the resulting salt was filtered and concentrated by distillation under reduced pressure. This concentrated solution was dissolved in chloroform and excess hexane was added to induce precipitation. This process was repeated 2-3 times, dried under reduced pressure, dissolved in distilled water, and precipitated by adding an appropriate amount of a substance (CF ₃ CH ₃ OH) lowering the low critical solution temperature. After centrifugation the sediment was freeze-dried, [NP (MPEG350) (LysOEt )] 3 1.7 g (yield, 70%). N-carbobenzyloxy-L-leusil-2- (5-fluorouracil-1-yl) -L, D-glycine (1.4 g, 3.2 mmol), N, N'-dicyclohexylcarbodiimide (0.66 g, 3.2 mmol) and 20 mL of dried tetrahydrofuran in N-hydroxysuccinimide (0.37 g, 3.2 mmol) were stirred for 24 hours, and then the resulting solution was dried in a tetrahydrofuran (20 mL) solvent. A solution of [NP (MPEG350) (LysOEt)] ₃ (1.7 g, 1.0 mmol) prepared above was added thereto and reacted at room temperature for 24 hours.

The reaction product was concentrated under reduced pressure, and the obtained material was dissolved in methanol, filtered, and then the unreacted material was removed using a dialysis membrane (MW, cutoff = 2,000). After dialysis for 48 hours, 10% palladium / charcoal catalyst (0.34 g) was added to the concentrated solution by distillation under reduced pressure (20 mL), and reacted under hydrogen gas for 4 to 5 hours. After distillation under reduced pressure to remove the solvent, it was dissolved in water. The solution obtained by filtration of only the part dissolved in water was lyophilized to obtain the final phosphazene trimer product {NP [O (CH ₂ CH ₂ O) ₇ CH ₃ ] [Lys-Gly (5-Fu) -Leu]} ₃ . 1.5 g (yield, 55%) were obtained.

Formula: C ₁₀₅ H ₁₈₉ N ₂₁ O ₄₂ F ₃ P ₃ ^. 6H ₂ O

Elemental Analysis Value: C, 46.7; H, 7. 38; N, 10.8

Theoretical: C, 46.6; H, 7.06; N, 10.9

Hydrogen Nuclear Magnetic Resonance Spectrum (Aceton-d ₆ , ppm):

δ 0.9 (b, 6H, -NHCOCHCH ₂ CH (C H ₃ ) ₂ NH ₂ ),

δ 1.5-1.9 (b, 12H, -NHCH ₂ (C H ₂ ) ₃ CHCO ₂ CH ₂ C H ₃ NH-,

-NHCOCHC H ₂ C H (CH ₃ ) ₂ NH ₂ ),

δ 2.9 (b, 2H, -NHC H ₂ (CH ₂ ) ₃ CHCO ₂ CH ₂ CH ₃ NH-),

δ 3.3 (s, 3H, -O (CH ₂ CH ₂ O) ₇ C H ₃ ),

δ 3.6-3.8 (b, 27H, -OCH ₂ C H ₂ O (C H ₂ C H ₂ O) ₆ CH _3,

-NHCOC H CH ₂ CH (CH ₃ ) ₂ NH ₂ ),

δ 4.0-4.3 (b, 4H, -OC H ₂ CH ₂ O (CH ₂ CH ₂ O) ₆ CH _3,

-NHCH ₂ (CH ₂ ) ₃ CHCO ₂ C H ₂ CH ₃ NH-),

δ 4.4 (b, 1H, -NHCH ₂ (CH ₂ ) ₃ C H CO ₂ CH ₂ CH ₃ NH-),

δ 6.4 (b, 1H, -NHCOC H (NCONHCOCFCH) NH-),

δ 8.2 (b, 1H, —NHCOCH (NCONHCOCFC H ) NH—).

Phosphorus nuclear magnetic resonance spectrum (Aceton-d ₆ , ppm): δ 38-44

HPLC: 99.82%

Low critical solution temperature: Not observed (above 100 ° C)

Example 2

[(Poly (methoxyethyleneglycol 350)] [L-phenylalanyl-2- (5-fluorouracil-1-yl) -L, D-glycyl-L-lysineethylester] cyclotriphosphazene, {NP [O (CH ₂ CH ₂ O) ₇ CH ₃ ] [Lys-Gly (5-Fu) -Phe]} ₃ Manufacture

Poly (methoxyethylene glycol) ethanol (1.8 g, 5.0 mmol), molecular weight 350 (0.14 g, 6.0 mmol), hexacyclotriphosphazene (0.50 g, 1.4 mmol), lysine ethyl ester (3.0 g) , 17 mmol), triethylamine (1.2 mL, 8.4 mmol), N-carbenzyloxy-L-phenylalanyl-2- (5-fluorouracil-1-yl) -L, D-glycine (1.5 g , 3.2 mmol), N, N'-dicyclohexylcarbodiimide (0.66 g, 3.2 mmol), N-hydroxysuccinimide (0.37 g, 3.2 mmol) and 10% palladium / charcoal catalyst (0.34 g) 1.4 g (yield, 50%) of the final phosphazene trimer product {NP [O (CH ₂ CH ₂ O) ₇ CH ₃ ] [Lys-Gly (5-Fu) -Phe]} _{3 in the} same manner as in Example 1 )

Formula: C ₁₁₅ H ₁₈₃ N ₂₁ O ₄₂ F ₃ P ₃ ^. 6H ₂ O

Elemental Analysis Value: C, 49.0; H, 7.00; N, 9.62

Theoretical: C, 49.2; H, 7.02; N, 10.5

Hydrogen Nuclear Magnetic Resonance Spectrum (D ₂ O, ppm)

δ 1.2-1.9 (b, 9H, -NHCH ₂ (C H ₂ ) ₃ CHCO ₂ CH ₂ C H ₃ NH-),

δ 2.9 (b, 2H, -NHC H ₂ (CH ₂ ) ₃ CHCO ₂ CH ₂ CH ₃ NH-),

δ 3.2 (b, 2H, -NHCOCHC H ₂ C ₆ H ₅ NH ₂ ),

δ 3.3 (s, 3H, -O (CH ₂ CH ₂ O) ₇ C H ₃ ),

δ 3.6-3.8 (b, 27H _, -OCH ₂ C H ₂ O (C H ₂ C H ₂ O) ₆ CH _3, -NHCOC H CH ₂ C ₆ H ₅ NH ₂ ),

δ 4.0-4.2 (b, 4H, -OC H ₂ CH ₂ O (CH ₂ CH ₂ O) ₆ CH _3,

-NHCH ₂ (CH ₂ ) ₃ CHCO ₂ C H ₂ CH ₃ NH-),

δ 4.3 (b, 1H, -NHCH ₂ (CH ₂ ) ₃ C H CO ₂ CH ₂ CH ₃ NH-),

δ 6.4 (b, 1H, -NHCOC H (NCONHCOCFCH) NH-),

δ 7.3 (b, 5H, -NHCOCHCH ₂ C ₆ H ₅ NH ₂ ),

δ 7.6-7.8 (b, 1H, -NHCOCH (NCONHCOCFC H ) NH-).

Phosphorus nuclear magnetic resonance spectrum (D ₂ O, ppm): δ 41.76

HPLC: 99.71%

Low critical solution temperature: 56 ℃

Example 3

[(2- (2'-methoxyethoxy) ethoxy] [L-leusil-2- (5-fluorouracil-1-yl) -L, D-glysil-L-lysineethylester] cyclo Triphosphazene, {NP [O (CH ₂ CH ₂ O) ₂ CH ₃ ] [Lys-Gly (5-Fu) -Leu]} ₃ Manufacture

2- (2'-methoxyethoxy) ethanol (0.6 g, 5.0 mmol), sodium metal chips (0.14 g, 6.0 mmol), hexacyclotriphosphazene (0.50 g, 1.4 mmol), lysine ethyl ester (3.0 g, 17 mmol) and triethylamine (1.2 mL, 8.4 mmol) in the same manner as in Example 1 {NP [O (CH ₂ CH ₂ O) ₂ CH ₃ ] [LysOEt]} ₃ 1.1 g (yield) , 79%). N-carbenzyloxy-L-leusil-2- (5-fluorouracil-1-yl) -L, D-glycine (1.6 g, 3.5 mmol), N, N'-dicyclohexylcarbodiimide (0.72 g, 3.5 mmol), N-hydroxysuccinimide (0.40 g, 3.5 mmol), 10% palladium / charcoal catalyst (0.33 g) and the final phosphazene trimer product {NP [ 0.92 g (yield, 43%) of O (CH ₂ CH ₂ O) ₂ CH ₃ ] [Lys-Gly (5-Fu) -Leu]} _{3 was} obtained.

Formula: C ₇₅ H ₁₂₉ N ₂₁ O ₂₇ F ₃ P ₃ ^. 6H ₂ O

Elemental Analysis Value: C, 45.6; H, 6. 86; N, 14.6

Theoretical: C, 45.5; H, 7.00; N, 14.9

Hydrogen Nuclear Magnetic Resonance Spectrum (D ₂ O, ppm)

δ 0.9 (b, 6H, -NHCOCHCH ₂ CH ( CH ₃ ) ₂ NH ₂ ),

δ 1.2 (b, 3H, -NHCH ₂ (CH ₂ ) ₃ CHCO ₂ CH ₂ C H ₃ NH-),

δ 1.4-1.9 (b, 9H, -NHCH ₂ (C H ₂ ) ₃ CHCO ₂ CH ₂ CH ₃ NH-,

-NHCOCHC H ₂ C H (CH ₃ ) ₂ NH ₂ ),

δ 2.9 (b, 2H, -NHC H ₂ (CH ₂ ) ₃ CHCO ₂ CH ₂ CH ₃ NH-),

δ 3.3 (s, 3H, —O (CH ₂ CH ₂ O) ₂ C H ₃ ),

δ 3.6-3.8 (b, 7H, -OCH ₂ C H ₂ OC H ₂ C H ₂ OCH _3, -NHCOC H CH ₂ CH (CH ₃ ) ₂ NH ₂ ),

δ 4.0 (b, 2H, -OC H ₂ CH ₂ OCH ₂ CH ₂ OCH ₃ ),

δ 4.2 (b, 2H, -NHCH ₂ (CH ₂ ) ₃ CHCO ₂ C H ₂ CH ₃ NH-),

δ 4.4 (b, 1H, -NHCH ₂ (CH ₂ ) ₃ C H CO ₂ CH ₂ CH ₃ NH-),

δ 6.5 (b, 1H, -NHCOC H (NCONHCOCFCH) NH-),

δ 7.8 (b, 1H, -NHCOCH (NCONHCOCFC H ) NH-).

Phosphorus Nuclear Magnetic Resonance Spectrum (D ₂ O, ppm): δ 42.30

HPLC: 99.95%

Low critical solution temperature: 27 ℃

Example 4

[(2- (2'-methoxyethoxy) ethoxy] [L-phenylalanyl-2-5-fluorouracil-1-yl-L, D-glycyl-L-lysineethylester] cyclotree Phosphazene, {NP [O (CH ₂ CH ₂ O) ₂ OCH ₃ ] [Lys-Gly (5-Fu) -Phe]} ₃ Manufacture

2- (2'-methoxyethoxy) ethanol (0.6 g, 5.0 mmol), sodium metal chips (0.14 g, 6.0 mmol), hexacyclotriphosphazene (0.50 g, 1.4 mmol), lysine ethyl ester (3.0 g, 17 mmol), triethylamine (1.2 mL, 8.4 mmol), N-carbenzyloxy-L-phenylalanyl-2- (5-fluorouracil-1-yl) -L, D-glycine (1.7 g, 3.5 mmol), N, N'-dicyclohexylcarbodiimide (0.72 g, 3.5 mmol), N-hydroxysuccinimide (0.40 g, 3.5 mmol) and 10% palladium / charcoal catalyst (0.33 g) 0.97 g (yield, 41) of the final phosphazene trimer product {NP [O (CH ₂ CH ₂ O) ₂ CH ₃ ] [Lys-Gly (5-Fu) -Phe]} ₃ %) Obtained.

Formula: C ₈₄ H ₁₂₃ N ₂₁ O ₂₇ F ₃ P ₃ ^. 9H ₂ O

Elemental analysis: C, 46.5; H, 5.98; N, 13.1

Theoretical: C, 46.5; H, 6.56; N, 13.6

Hydrogen Nuclear Magnetic Resonance Spectrum (D ₂ O, ppm)

δ 1.2-1.9 (b, 9H, -NHCH ₂ (C H ₂ ) ₃ CHCO ₂ CH ₂ C H ₃ NH-),

δ 2.9 (b, 2H, -NHC H ₂ (CH ₂ ) ₃ CHCO ₂ CH ₂ CH ₃ NH-),

δ 3.2 (b, 2H, -NHCOCHC H ₂ C ₆ H ₅ NH ₂ ),

δ 3.3 (s, 3H, —O (CH ₂ CH ₂ O) ₂ C H ₃ ),

δ 3.6-3.8 (b, 7H, -OCH ₂ C H ₂ OC H ₂ C H ₂ OCH _3, -NHCOC H CH ₂ C ₆ H ₅ NH ₂ ),

δ 4.1 (b, 2H, -OC H ₂ CH ₂ OCH ₂ CH ₂ OCH ₃ ),

δ 4.2 (b, 2H, -NHCH ₂ (CH ₂ ) ₃ CHCO ₂ C H ₂ CH ₃ NH-),

δ 4.4 (b, 1H, -NHCH ₂ (CH ₂ ) ₃ C H CO ₂ CH ₂ CH ₃ NH-),

δ 6.4 (b, 1H, -NHCOC H (NCONHCOCFCH) NH-),

δ 7.3 (b, 5H, -NHCOCHCH ₂ C ₆ H ₅ NH ₂ ),

δ 7.6-7.8 (b, 1H, -NHCOCH (NCONHCOCFC H ) NH-)

Phosphorus Nuclear Magnetic Resonance Spectrum (D ₂ O, ppm): δ 42.37

HPLC: 100%

Low critical solution temperature: 11 ℃

Example 5

[(2- (2'-methoxyethoxy) ethoxy] [L-alanyl-2- (5-fluorouracil-1-yl) -L, D-glysil-L-lysineethylester] cyclo Triphosphazene, {NP [(OCH ₂ CH ₂ ) ₂ OCH ₃ ] [Lys-Gly (5-Fu) -Ala]} ₃ Manufacture

2- (2'-methoxyethoxy) ethanol (0.6 g, 5.0 mmol), sodium metal chips (0.14 g, 6.0 mmol), hexacyclotriphosphazene (0.50 g, 1.4 mmol), lysine ethyl ester (3.0 g, 17 mmol), triethylamine (1.2 mL, 8.4 mmol), N-carbenzyloxy-L-alanyl-2- (5-fluorouracil-1-yl) -L, D-glycine (1.4 g , 3.5 mmol), N, N'-dicyclohexylcarbodiimide (0.72 g, 3.5 mmol), N-hydroxysuccinimide (0.40 g, 3.5 mmol), 10% palladium / charcoal catalyst (0.33 g) 0.89 g (Yield, 41%) of the final phosphazene trimer product {NP [O (CH ₂ CH ₂ O) ₂ CH ₃ ] [Lys-Gly (5-Fu) -Ala]} ₃ )

Formula: C ₆₆ H ₁₁₁ N ₂₁ O ₂₇ F ₃ P ₃ ^. 11H ₂ O

Elemental Analysis Value: C, 39.8; H, 6. 16; N, 14.4

Theoretical: C, 40.1; H, 6.79; N, 14.9

Hydrogen Nuclear Magnetic Resonance Spectrum (D ₂ O, ppm)

δ 1.2 (b, 3H, -NHCOCH CH ₃ NH ₂ ),

δ 1.4-1.9 (b, 9H, -NHCH ₂ (C H ₂ ) ₃ CHCO ₂ CH ₂ C H ₃ NH-),

δ 2.9 (b, 2H, -NHC H ₂ (CH ₂ ) ₃ CHCO ₂ CH ₂ CH ₃ NH-),

δ 3.3 (s, 3H, —O (CH ₂ CH ₂ O) ₂ C H ₃ ),

δ 3.6-3.8 (b, 7H, -OCH ₂ C H ₂ OC H ₂ C H ₂ OCH _3, -NHCOC H CH ₃ NH ₂ ),

δ 4.0 (b, 2H, -OC H ₂ CH ₂ OCH ₂ CH ₂ OCH ₃ ),

δ 4.2 (b, 2H, -NHCH ₂ (CH ₂ ) ₃ CHCO ₂ C H ₂ CH ₃ NH-),

δ 4.4 (b, 1H, -NHCH ₂ (CH ₂ ) ₃ C H CO ₂ CH ₂ CH ₃ NH-),

δ 6.5 (b, 1H, -NHCOC H (NCONHCOCFCH) NH-),

δ 7.9 (b, 1H, -NHCOCH (NCONHCOCFC H ) NH-).

Phosphorus nuclear magnetic resonance spectrum (D ₂ O, ppm): δ 42.35

HPLC: 100%

Low critical solution temperature: not observed (above 100 ℃)

Example 6

[(2- (2'-Ethoxyethoxy) ethoxy] [L-Louisyl-2- (5-fluorouracil-1-yl) -L, D-glysil-L-lysineethylester] cyclo Triphosphazene, {NP [O (CH ₂ CH ₂ O) ₂ CH ₂ CH ₃ ] [Lys-Gly (5-Fu) -Leu]} ₃ Manufacture

2- (2'-ethoxyethoxy) ethanol (0.67 g, 5.0 mmol), sodium metal fragment (0.14 g, 6.0 mmol), hexacyclotriphosphazene (0.50 g, 1.4 mmol), lysine ethyl ester (3.0 g, 17 mmol) and triethylamine (1.2 mL, 8.4 mmol) gave 1.3 g (yield, 86%) of {NP [O (CH ₂ CH ₂ O) ₂ CH ₂ CH ₃ ] [LysOEt]} ₃ . . N-carbenzyloxy-L-leusil-2- (5-fluorourasil-1-yl) -L, D-glycine (1.7 g, 3.8 mmol), N, N'-dicyclohexylcarbodiimide (0.78 g, 3.8 mmol), N-hydroxysuccinimide (0.44 g, 3.8 mmol), 10% palladium / charcoal catalyst (0.39 g), and the final phosphazene trimer product {NP [ 1.1 g (yield, 43%) of O (CH ₂ CH ₂ O) ₂ CH ₂ CH ₃ ] [Lys-Gly (5-Fu) -Leu]} ₃ were obtained.

Formula: C ₇₈ H ₁₃₅ N ₂₁ O ₂₇ F ₃ P ₃ ^. 10H ₂ O

Elemental Analysis Value: C, 43.9; H, 6.83; N, 13.2

Theoretical: C, 44.0; H, 7. 35; N, 13.8

Hydrogen Nuclear Magnetic Resonance Spectrum (D ₂ O, ppm)

δ 0.9 (b, 6H, -NHCOCHCH ₂ CH (C H ₃ ) ₂ NH ₂ ),

δ 1.2 (b, 6H, —O (CH ₂ CH ₂ O) ₂ CH ₂ C H _3, -NHCH ₂ (CH ₂ ) ₃ CHCO ₂ CH ₂ C H ₃ NH-),

δ 1.5-1.9 (b, 9H, -NHCH ₂ (C H ₂ ) ₃ CHCO ₂ CH ₂ CH ₃ NH-,

-NHCOCHC H ₂ C H (CH ₃ ) ₂ NH ₂ ),

δ 2.9 (b, 2H, -NHC H ₂ (CH ₂ ) ₃ CHCO ₂ CH ₂ CH ₃ NH-),

δ 3.3 (s, 3H, —O (CH ₂ CH ₂ O) ₂ CH ₂ C H ₃ ),

δ 3.6-3.8 (b, 9H, -OCH ₂ C H ₂ OC H ₂ C H ₂ OC H ₂ CH _3,

-NHCOC H CH ₂ CH (CH ₃ ) ₂ NH ₂ ),

δ 4.0 (b, 2H, -OC H ₂ CH ₂ OCH ₂ CH ₂ OCH ₂ CH ₃ ),

δ 4.2 (b, 2H, -NHCH ₂ (CH ₂ ) ₃ CHCO ₂ C H ₂ CH ₃ NH-),

δ 4.4 (b, 1H, -NHCH ₂ (CH ₂ ) ₃ C H CO ₂ CH ₂ CH ₃ NH-),

δ 6.5 (b, 1H, -NHCOC H (NCONHCOCFCH) NH-),

δ 7.9 (b, 1H, -NHCOCH (NCONHCOCFC H ) NH-).

Phosphorus nuclear magnetic resonance spectrum (D ₂ O, ppm): δ 42.18

HPLC: 99.96%

Low critical solution temperature: 17 ℃

Example 7

[(2- (2'-Ethoxyethoxy) ethoxy] [L-valyl-2- (5-fluorouracil-1-yl) -L, D-glycyl-L-lysineethylester] cyclotree Phosphazene, {NP [O (CH ₂ CH ₂ O) ₂ CH ₂ CH ₃ ] [Lys-Gly (5- Fu) -Val]} ₃ Manufacture

2- (2'-ethoxyethoxy) ethanol (0.67 g, 5.0 mmol), sodium metal fragment (0.14 g, 6.0 mmol), hexacyclotriphosphazene (0.50 g, 1.4 mmol), lysine ethyl ester (3.0 g, 17 mmol), triethylamine (1.2 mL, 8.4 mmol), N-carbenzyloxy-L-valyl-2- (5-fluorouracil-1-yl) -L, D-glycine (1.7 g, 3.8 mmol), N, N'-dicyclohexylcarbodiimide (0.78 g, 3.8 mmol), N-hydroxysuccinimide (0.44 g, 3.8 mmol), 10% palladium / charcoal catalyst (0.39 g) In the same manner as in Example 1, the final phosphazene trimer product {NP [O (CH ₂ CH ₂ O) ₂ CH ₂ CH ₃ ] [Lys-Gly (5-Fu) -Val]} ₃ 1.3 g (yield, 56 %) Obtained.

Formula: C ₇₅ H ₁₂₉ N ₂₁ O ₂₇ F ₃ P ₃ ^. 2H ₂ O

Elemental Analysis Value: C, 46.1; H, 6.87; N, 14.2

Theoretical: C, 46.4; H, 6.91; N, 15.1

Hydrogen Nuclear Magnetic Resonance Spectrum (D ₂ O, ppm)

δ 0.9 (b, 6H, -NHCOCHCH (C H ₃ ) ₂ NH ₂ ),

δ 1.2 (b, 6H, —O (CH ₂ CH ₂ O) ₂ CH ₂ C H _3, -NHCH ₂ (CH ₂ ) ₃ CHCO ₂ CH ₂ C H ₃ NH-),

δ 1.5-1.9 (b, 7H, -NHCH ₂ (C H ₂ ) ₃ CHCO ₂ CH ₂ CH ₃ NH-,

-NHCOCHC H (CH ₃ ) ₂ NH ₂ ),

δ 2.9 (b, 2H, -NHC H ₂ (CH ₂ ) ₃ CHCO ₂ CH ₂ CH ₃ NH-),

δ 3.3 (s, 3H, —O (CH ₂ CH ₂ O) ₂ CH ₂ C H ₃ ),

δ 3.6-3.8 (b, 9H, -OCH ₂ C H ₂ OC H ₂ C H ₂ OC H ₂ CH _3, -NHCOC H CH (CH ₃ ) ₂ NH ₂ ),

δ 4.0 (b, 2H, -OC H ₂ CH ₂ OCH ₂ CH ₂ OCH ₂ CH ₃ ),

δ 4.2 (b, 2H, -NHCH ₂ (CH ₂ ) ₃ CHCO ₂ C H ₂ CH ₃ NH-),

δ 4.4 (b, 1H, -NHCH ₂ (CH ₂ ) ₃ C H CO ₂ CH ₂ CH ₃ NH-),

δ 6.5 (b, 1H, -NHCOC H (NCONHCOCFCH) NH-),

δ 7.9 (b, 1H, -NHCOCH (NCONHCOCFC H ) NH-)

Phosphorus nuclear magnetic resonance spectrum (D ₂ O, ppm): δ 42.18

HPLC: 99.94%

Low critical solution temperature: 15 ℃

Example 8

[(2- (2'-butoxyethoxy) ethoxy] [L-alanyl-2- (5-fluorouracil-1-yl) -L, D-glysil-L-lysineethylester] cyclo Triphosphazene, {NP [O (CH ₂ CH ₂ O) ₂ CH ₂ CH ₂ CH ₂ CH ₃ ] [Lys-Gly (5-Fu) -Ala]} ₃ Manufacture

2- (2'-butoxyethoxy) ethanol (0.82 g, 5.0 mmol), sodium metal chips (0.14 g, 6.0 mmol), hexacyclotriphosphazene (0.50 g, 1.4 mmol), lysine ethyl ester (3.0 g, 17 mmol) and triethylamine (1.2 mL, 8.4 mmol) in the same manner as in Example 1 {NP [O (CH ₂ CH ₂ O) ₂ CH ₂ CH ₂ CH ₂ CH ₃ ] [LysOEt] } ₃ 1.1 g (yield, 69%) were obtained. N-Cabobenzyloxy-L-alanyl-2- (5-fluorouracil-1-yl) -L, D-glycine (1.4 g, 3.4 mmol), N, N'-dicyclohexylcarbodiimide (0.70 g, 3.4 mmol), N-hydroxysuccinimide (0.39 g, 3.4 mmol), 10% palladium / charcoal catalyst (0.33 g) and the final phosphazene trimer product {NP [ 0.92 g (yield, 46%) of O (CH ₂ CH ₂ O) ₂ CH ₂ CH ₂ CH ₂ CH ₃ ] [Lys-Gly (5-Fu) -Ala]} _{3 was} obtained.

Formula: C ₇₅ H ₁₂₉ N ₂₁ O ₂₇ F ₃ P ₃ ^. 7H ₂ O

Elemental Analysis Value: C, 44.4; H, 6.68; N, 13.9

Theoretical: C, 44.3; H, 7. 10; N, 14.5

Hydrogen Nuclear Magnetic Resonance Spectrum (D ₂ O, ppm)

δ 0.9 (b, 3H, -O (CH ₂ CH ₂ O) ₂ CH ₂ CH ₂ CH ₂ C H ₃ ),

δ 1.2-1.9 (b, 16H, -NHCH ₂ (C H ₂ ) ₃ CHCO ₂ CH ₂ C H ₃ NH-,

-O (CH ₂ CH ₂ O) ₂ CH ₂ C H ₂ C H ₂ CH _3, -NHCOCHC H ₃ NH ₂ ),

δ 2.9 (b, 2H, -NHC H ₂ (CH ₂ ) ₃ CHCO ₂ CH ₂ CH ₃ NH-),

δ 3.5 (s, 2H, —O (CH ₂ CH ₂ O) ₂ C H ₂ CH ₂ CH ₂ CH ₃ ),

δ 3.6-3.8 (b, 7H, -OCH ₂ C H ₂ OC H ₂ C H ₂ OCH ₂ CH ₂ CH ₂ CH _3,

-NHCOC H CH ₃ NH ₂ ),

δ 4.0 (b, 2H, -OC H ₂ CH ₂ OCH ₂ CH ₂ OCH ₂ CH ₂ CH ₂ CH ₃ ),

δ 4.2 (b, 2H, -NHCH ₂ (CH ₂ ) ₃ CHCO ₂ C H ₂ CH ₃ NH-),

δ 4.4 (b, 1H, -NHCH ₂ (CH ₂ ) ₃ C H CO ₂ CH ₂ CH ₃ NH-),

δ 6.5 (b, 1H, -NHCOC H (NCONHCOCFCH) NH-),

δ 7.9 (b, 1H, -NHCOCH (NCONHCOCFC H ) NH-).

Phosphorus Nuclear Magnetic Resonance Spectrum (D ₂ O, ppm): δ 42.12

HPLC: 98.85%

Low critical solution temperature: 14 ℃

Example 9

[(2- (2'-butoxyethoxy) ethoxy] [L-glycyl-2- (5-fluorouracil-1-yl) -L, D-glycyl-L-lysineethylester] cyclo Triphosphazene, {NP [O (CH ₂ CH ₂ O) ₂ CH ₂ CH ₂ CH ₂ CH ₃ ] [Lys-Gly (5-Fu) -Gly]} ₃ Manufacture

2- (2'-butoxyethoxy) ethanol (0.82 g, 5.0 mmol), sodium metal chips (0.14 g, 6.0 mmol), hexacyclotriphosphazene (0.50 g, 1.4 mmol), lysine ethyl ester (3.0 g, 17 mmol), triethylamine (1.2 mL, 8.4 mmol), N-carbenzyloxy-L-alanyl-2- (5-fluorouracil-1-yll) -L, D-glycine (1.3 g, 3.4 mmol), N, N'-dicyclohexylcarbodiimide (0.70 g, 3.4 mmol), N-hydroxysuccinimide (0.39 g, 3.4 mmol) and 10% palladium / charcoal catalyst (0.33 g) Final phosphazene trimer product {NP [O (CH ₂ CH ₂ O) ₂ CH ₂ CH ₂ CH ₂ CH ₃ ] [Lys-Gly (5-Fu) -Gly]} in the same manner as in Example 1 ₃ 0.87 g (yield, 45%) was obtained.

Formula: C ₇₂ H ₁₂₃ N ₂₁ O ₂₇ F ₃ P ₃ ^. 7H ₂ O

Elemental Analysis Value: C, 43.8; H, 6.57; N, 14.4

Theoretical: C, 43.4; H, 6. 95; N, 14.8

Hydrogen Nuclear Magnetic Resonance Spectrum (D ₂ O, ppm)

δ 0.9 (b, 3H, -O (CH ₂ CH ₂ O) ₂ CH ₂ CH ₂ CH ₂ C H ₃ ),

δ 1.2-1.9 (b, 13H, -NHCH ₂ (C H ₂ ) ₃ CHCO ₂ CH ₂ C H ₃ NH-,

-O (CH ₂ CH ₂ O) ₂ CH ₂ C H ₂ C H ₂ CH ₃ ),

δ 2.9 (b, 2H, -NHC H ₂ (CH ₂ ) ₃ CHCO ₂ CH ₂ CH ₃ NH-),

δ 3.5 (s, 2H, —O (CH ₂ CH ₂ O) ₂ C H ₂ CH ₂ CH ₂ CH ₃ ),

δ 3.6-3.8 (b, 6H, -OCH ₂ C H ₂ OC H ₂ C H ₂ OCH ₂ CH ₂ CH ₂ CH ₃ ),

δ 3.9 (b, 2H, -NHCOC H ₂ NH ₂ ),

δ 4.0 (b, 2H, -OC H ₂ CH ₂ OCH ₂ CH ₂ OCH ₂ CH ₂ CH ₂ CH ₃ ),

δ 4.2 (b, 2H, -NHCH ₂ (CH ₂ ) ₃ CHCO ₂ C H ₂ CH ₃ NH-),

δ 4.4 (b, 1H, -NHCH ₂ (CH ₂ ) ₃ C H CO ₂ CH ₂ CH ₃ NH-),

δ 6.5 (b, 1H, -NHCOC H (NCONHCOCFCH) NH-),

δ 7.9 (b, 1H, -NHCOCH (NCONHCOCFC H ) NH-).

Phosphorus Nuclear Magnetic Resonance Spectrum (D ₂ O, ppm): δ 42.25

HPLC: 99.86%

Low critical solution temperature: 19 ℃

Experimental Example Biological Activity Test and Results of Platinum Complex

Toxicity and physiological activity tests on the phosphazene trimer product obtained in the Examples of the present invention were performed as follows.

One. In vitro  Drug search

(1) Sample: The phosphazene trimer product prepared in Example 1, Example 2, Example 3, Example 4, Example 5, Example 6, Example 7, Example 8, and Example 9.

(2) tumor cell line: L1210

(3) Experimental Method: L1210 cells cultured in RPMI 1640 medium containing 10% FBS were adjusted to a concentration of 1 × 10 ⁵ cells / ml. Drugs of each concentration diluted in log dose were added to the cells, and cultured in a 37 ° C., 5% CO ₂ incubator, and after 24, 48, and 72 hours, each viable cell number Was measured. Live cell counts were subjected to a dye exclusion test using tryptophan blue. From the measured cell number, the concentration of each compound (IC ₅₀ ) showing 50% cell proliferation inhibition was compared with that of the control group, and the results are shown in Table 1.

2. In vivo (i.p.)  Drug search

(1) Sample: The phosphazene trimer product prepared in Examples 1, 2, 3 and 5.

(2) tumor cell line: mouse leukemia L1210

(3) Experimental Method: Eight 6-week-old BDF1 mice were transplanted into groups of 1 × 10 ⁵ cells / 0.1 ml of leukemia L1210 cells, which were DBA / 2 mouse masses, into each group. The test drug is dissolved in PBS (-) or suspended in 0.5% Tween 80, depending on the drug, administered continuously from 1 day to 5 days after cancer implantation or injected intraperitoneally at respective concentrations on 1, 5 and 9 days. It was. Survival was measured while the mice were observed daily, and from the measured average survival time of each experimental group, an increased ratio (T / C (%)) of the average survival days of the administration group compared to the control group was determined to determine the anticancer effect. The results are shown in Table 1.

Table 1 Tests and results of physiological activity of phosphazene trimer-5-fluorouracil complex

Compound class In vitro (ED ₅₀ ) In vivo (ip) μg / mL μM Dose (mg / kg) T / C (%) Example 1 4.2 1.6 150 113.2 Example 2 7.3 2.6 150 104.1 Example 3 1.7 0.9 150 154.7 Example 4 7.5 3.5 Example 5 2.0 1.0 150 142.4 Example 6 4.0 2.1 Example 7 7.0 3.3 Example 8 3.7 1.9 Example 9 5.4 2.9 5-fluorouracil (control) 0.3 4.6 25 186.8

The present invention provides a cyclic phosphazene trimer-5-fluorouracil complex anticancer system having a stereoselective chemical structure and introducing a temperature sensitive concept. The cyclic phosphazene trimer-5-fluorouracil complex system of the present invention has the temperature sensitivity, and has the advantage that the temperature can be easily and accurately controlled. Therefore, the temperature sensitive cyclic phosphazene trimer-5-fluorouracil complex anticancer agent of the present invention is expected to be applied to anticancer treatment including systemic administration and local administration in the future.

Claims (11)

A temperature sensitive cyclic phosphazene trimer-5-fluorouracil complex having the structure of formula (I)

Wherein m is an integer selected from 2 and 7, n is 0 or an integer selected from 1, 2, and 3, R is H, CH ₃ , CH (CH ₃ ) ₂ , CH ₂ CH (CH ₃ ) ₂ and CH ₂ C ₆ H ₆ is selected from the group consisting of. After reacting hexacyclotriphosphazene with a hydrophilic poly (alkoxyethylene glycol), a nucleophilic substitution reaction is carried out using an excess of lysine ethyl ester to give a cyclic phosphazene trimer intermediate compound having the structure of formula (II). To manufacture; A compound having the structure of formula (VI) is prepared by reacting a compound having the structure of formula (III), a compound having the structure of formula (IV), and a compound having the structure of formula (V). and: Reacting the intermediate compound having the structure of the general formula (II) with the compound having the structure of the general formula (VI) to prepare a compound having the structure of the following general formula (VII); Reducing the compound of formula (VII) obtained above using a palladium / charcoal catalyst under hydrogen gas to prepare a temperature sensitive cyclic phosphazene trimer-5-fluorouracil complex having the structure of formula (I). Including, Process for preparing a temperature sensitive cyclic phosphazene trimer-5-fluorouracil complex having a structure of formula (I) according to claim 1:

Wherein m is an integer selected from 2 and 7, n is 0 or an integer selected from 1, 2, and 3, R is H, CH ₃ , CH (CH ₃ ) ₂ , CH ₂ CH (CH ₃ ) ₂ and CH ₂ C ₆ H ₆ is selected from the group consisting of. The method according to claim 1, wherein the hexacyclotriphosphazene and the hydrophilic poly (alkoxyethylene glycol) are 1: 2.8 to 1: in the step of preparing a cyclic phosphazene trimer intermediate compound having the structure of Formula (II). 3.2 (hexacyclotriphosphazene: hydrophilic poly (alkoxyethyleneglycol)) reaction in a molar ratio. The method according to claim 1, wherein in the step of preparing a cyclic phosphazene trimer intermediate compound having the structure of Formula (II), the compound having the structure of Formula (II) is dissolved in a small amount of chloroform, and then And adding hexane to precipitate the final product to increase the purity of the compound having the structure of formula (II). The method according to claim 4, wherein in the step of preparing a cyclic phosphazene trimer intermediate compound having the structure of Formula (II), the precipitate is dissolved in distilled water and CF ₃ CH ₂ OH is a substance which lowers the low critical temperature. The method further comprises the step of inducing the precipitate in an amount that can be induced to precipitate, followed by centrifugation to obtain a compound having a structure of the general formula (II) with increased purity. The compound according to claim 1, wherein in the step of preparing a compound having the structure of general formula (VI), the compound having the structure of general formula (III), the compound having the structure of general formula (IV) and the general formula (V) A compound having a structure is reacted in a molar ratio of 1: 1: 1. The compound according to claim 1, wherein in the step of preparing the compound having the structure of Formula (VII), the intermediate compound having the structure of Formula (II) is combined with the compound having the structure of Formula (VI): It is made to react by the molar ratio of 3-1: 4 [compound of general formula (II): compound of general formula (VI)]. The preparation according to claim 7, further comprising dissolving the obtained product in methanol and removing unreacted substances and by-products using a dialysis membrane (MW, cutoff = 2,000) to obtain a more pure compound of formula (VII). Way. The method of claim 1, wherein in the step of reducing the compound of general formula (VII) to prepare a compound of general formula (I), the obtained product is dissolved in methol, and then hydrogenated using a 10% palladium / charcoal catalyst. After reacting for 4 to 5 hours under gas, the obtained product is dissolved in water, and only the dissolved portion is lyophilized to prepare a compound of general formula (I) as a final product. A cell proliferation inhibitor containing an effective amount of the temperature sensitive cyclic phosphazene trimer-5-fluorouracil complex according to claim 1 as an active ingredient. An anticancer agent containing an effective amount of the temperature sensitive cyclic phosphazene trimer-5-fluorouracil complex according to claim 1 as an active ingredient.