KR980009276A - PVMA-Ara-C conjugate - Google Patents

PVMA-Ara-C conjugate Download PDF

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KR980009276A
KR980009276A KR1019960028315A KR19960028315A KR980009276A KR 980009276 A KR980009276 A KR 980009276A KR 1019960028315 A KR1019960028315 A KR 1019960028315A KR 19960028315 A KR19960028315 A KR 19960028315A KR 980009276 A KR980009276 A KR 980009276A
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Abstract

본 발명은 PVMA-Ara-C 결합체에 관한 것으로서, 더욱 상세하기로는 한번 투여로 장시간 약물 혈중농도를 지속시켜 항암효과를 증진시키는 다음 구조식(Ⅰ)로 표시되는 PVMA-Ara-C 결합체, 이의 제조방법 및 이를 유효성분으로 함유하는 항암제에 관한 것이다.The present invention relates to a PVMA-Ara-C conjugate, and more particularly, to a PVMA-Ara-C conjugate represented by the following structural formula (I) for enhancing an anticancer effect by sustaining a blood drug concentration for a long time by single administration, And an anticancer agent containing the same as an active ingredient.

상기 식에서 x와 y의 비는 0:100∼80:20이다.Wherein the ratio of x and y is from 0: 100 to 80: 20.

Description

PVMA-Ara-C 결합체PVMA-Ara-C conjugate

본 발명은 PVMA-Ara-C 결합체에 관한 것으로서, 더욱 상세하기로는 한번 투여로 장시간 약물 혈중농도를 지속시켜 항암효과를 증진시키는 다음 구조식(Ⅰ)로 표시되는 PVMA-Ara-C 결합체, 이의 제조방법 및 이를 유효성분으로 함유하는 항암제에 관한 것이다.The present invention relates to a PVMA-Ara-C conjugate, and more particularly, to a PVMA-Ara-C conjugate represented by the following structural formula (I) for enhancing an anticancer effect by sustaining a blood drug concentration for a long time by single administration, And an anticancer agent containing the same as an active ingredient.

상기 식에서, x와 y의 비는 0:100∼80:20이다.In the above formula, the ratio of x and y is 0: 100 to 80: 20.

아라비노프라노실시토신(1-β-D-Arabinofuranosylcytosine; 이하, "Ara-C" 라 함)은 세포의 DNA합성을 차단하여 암세포를 죽이는 메카니즘을 보여주며 암상적으로 백혈병 치료에 이용되고 있으나, 혈청 또는 조직에서 디옥시시티딘 디아미나제(deoxycytidine deaminase)에 의하여 우라실 유사체로 바뀌어져 항암 작용을 할 수 없게 되고, 또한 최대약물을 주사하여도 신장 여과작용에 의하여 약물이 방출되기 때문에 가능한 한 약물을 연속적으로 오랜 시간동안 주사하는 것이 치료에 효과적이다.Arabinofuranosylcytosine (hereinafter referred to as "Ara-C") shows a mechanism of killing cancer cells by blocking DNA synthesis of cells, and is used for treating leukemia in a cancerous manner, Or deoxycytidine deaminase in the tissue, the drug can not be converted into the uracil analogue, and the drug is released by the renal filtration even when the maximum drug is injected. Therefore, Continuous long-term injections are effective for treatment.

Ara-C는 S기 암세포에 대해 활성을 가지기 때문에 혈중농도를 유효농도 이상으로 장시간 유지하는 것이 중요하다. [Cancer Chemotherapy Report V5l, 125, 1967). 그러나 장시간에 걸친 점적 주사방법은 환자에게 부담이 크기 때문에 주사방법이 아닌 새로운 약물 투여방법에 대한 연구가 진행되어 왔다. 이러한 연구결과로서, Ara-C를 리포좀에 함입시켜 얻은 제형이 개발되었는데, 리포좀은 효소의 비활성화로부터 Ara-C를 보호하고 약물을 서서히 방출시키는 서방형제제로 약물의 유효농도가 오랫동안 지속되어 항암 효과를 증진시키거나[Int. J. Cancer, V2O, 581, 1977], 리포좀이 혈액 중에서 불안정하여 쉽게 깨지는 문제가 있다.Since Ara-C has activity against S-phase cancer cells, it is important to maintain the blood concentration over the effective concentration for a long time. [Cancer Chemotherapy Report V5l, 125, 1967). However, since the long-term drip injection method is burdensome to the patient, research on a new drug administration method has been carried out instead of the injection method. As a result of these studies, a formulation has been developed that incorporates Ara-C into liposomes. Liposomes are Ara-C protectants from the inactivation of enzymes and slow release of the drug. The effective concentration of the drug is long lasting, Or [Int. J. Cancer, V2O, 581, 1977], liposomes are unstable in blood and are easily broken.

최근에는 항암제 전달체의 불안정성을 극복하기 위해 합성고분자를 이용한 약물전달 방법들이 이용되고 있는데, 합성 고분자에 항암제를 결합하여 얻은 고분자 약들의 경우 항암제가 안정하게 유지되고, 항암제가 서서히 방출되어 한번의 투여로 장시간 약물 혈중농도가 지속되기 때문에 항암 효과가 증진되는 우수성이 있다. [J. Controlled Release, Vl9, 331, 1992].In recent years, drug delivery methods using synthetic polymers have been used to overcome the instability of anticancer drug delivery systems. In the case of polymer drugs obtained by combining anticancer drugs with synthetic polymers, anticancer drugs are stably maintained, There is excellence in the improvement of anticancer effect because drug concentration in blood is maintained for a long time. [J. Controlled Release, Vl 9, 331, 1992].

본 발명에서는 항암효과가 우수한 Ara-C를 폴리피롤리돈-말레인산 무수물의 공중합체(이하 "PVMA"라 함)에 결합시켜 항암 효과를 증진시킨 신규 PVMA-Ara-C 결합체를 제공하는데 그 목적이 있다.It is an object of the present invention to provide a novel PVMA-Ara-C conjugate in which Ara-C having excellent anticancer effect is bound to a polypyrrolidone-maleic anhydride copolymer (hereinafter referred to as "PVMA") to enhance the anticancer effect.

본 발명은 다음 구조식(Ⅰ)로 표시되는 PVMA-Ara-C 결합체를 그 특징으로 한다.The present invention is characterized by the PVMA-Ara-C conjugate represented by the following structural formula (I).

상기 식에서, x와 y의 비는 0:100∼80:20이다.In the above formula, the ratio of x and y is 0: 100 to 80: 20.

또한, 본 발명은 상기 구조식(Ⅰ)로 표시되는 PVMA-Ara-C 결합체를 유효성분으로 함유하는 항암제를 포함한다.The present invention also includes an anticancer agent containing the PVMA-Ara-C conjugate represented by the structural formula (I) as an active ingredient.

또한, 본 발명은 다음 구조식(A)로 표시되는 PVMA를 친수성용매에 녹인 용액과 다음 구조식(B)로 표시되는 Ara-C를 브롬산 또는 인산 완충액에 녹인 용액을 중합반응시켜 상기 구조식(Ⅰ)로 표시되는 PVMA-Ara-C 결합체를 제조하는 방법을 포함한다.The present invention also relates to a process for producing a compound represented by the above formula (I) by polymerizing a solution obtained by dissolving PVMA represented by the following formula (A) in a hydrophilic solvent and a solution obtained by dissolving Ara-C represented by the following formula (B) in a bromic acid or a phosphate buffer, ≪ RTI ID = 0.0 > PVMA-Ara-C < / RTI >

상기 식에서, n은 10내지 500의 정수이다.In the above formula, n is an integer of 10 to 500.

이와 같은 발명을 더욱 상세히 설명하면 다음과 같다.This invention will be described in more detail as follows.

본 발명은 항암효과가 우수한 Ara-C와 PVMA의 결합에 의해 생성된 PVMA-Ara-C 결합체는 아미드 결합으로 이루어져 있으며, 인접한 카르복시기에 의한 뒤 공격(back-attack)효과에 의하여 생체내에서 효소에 관계없이 서서히 약물을 방출하므로 유효 혈중농도 이상으로 장시간 유지하게 되고 이로써 항암효과를 증진시키는 PVMA-Ara-C 결합체에 관한 것이다.The present invention is based on the finding that the PVMA-Ara-C conjugate formed by the combination of Ara-C and PVMA having excellent anticancer effect is composed of an amide bond, and is capable of inhibiting the enzymes in vivo by the back- The present invention relates to a PVMA-Ara-C conjugate capable of sustaining a drug over an effective blood concentration for a long period of time, thereby increasing the anticancer effect.

본 발명에서 Ara-C전달체로 사용하는 PVMA는 비닐피롤리돈 단량체와 말레산 무수물을 공지 방법[Makromol. Chem., 187, 297, 1986]에 의해 공중합시켜 제조한 고분자물질로서, 이는 생체적합성이 있고 수용성이기 때문에 고분자 약 개발에 매우 유망한 약물로 잘 알려져 있다. 또한, PVMA에는 화학적 반응성이 높은 다수의 말레산 무수물이 존재하기 때문에 아미노기(-NH2) 또는 수산기(-OH) 등의 활성수소 관능기를 가진 약물과 쉽게 높은 비율로 결합이 가능하고, 약물과의 반응 후 공중합체에 다수의 수용성의 카르복시기(-COOH)가 생성되기 때문에 생성된 PVMA-약물의 결합체는 매우 물에 잘 용해되는 특성이 있다. 본 발명에서는 반복단위(n)가 10∼500이고 분자량이 2,000∼100,000 범위인 PVMA를 사용하는 바, PVMA는 분자량에 관계없이 항암제와의 결합에 의해 항암증진 효과를 갖을 수 있지만 배설성 및 부작용을 고려하면 분자량 10,000 이하의 것을 사용하는 것이 바람직하다.In the present invention, the PVMA used as an Ara-C carrier is prepared by reacting a vinylpyrrolidone monomer and maleic anhydride with a known method [Makromol. Chem., 187, 297, 1986], which is a biocompatible and water-soluble polymer, is well known as a promising drug for the development of polymer drugs. Since PVMA has a large number of highly reactive maleic anhydrides, it can be easily coupled with a drug having an active hydrogen functional group such as an amino group (-NH 2 ) or a hydroxyl group (-OH) at a high rate, Since a large number of water-soluble carboxyl groups (-COOH) are produced in the copolymer after the reaction, the resulting PVMA-drug conjugate is highly water soluble. In the present invention, when PVMA having a repeating unit (n) of 10 to 500 and a molecular weight of 2,000 to 100,000 is used, PVMA can have anticancer-enhancing effect by binding with an anticancer drug regardless of the molecular weight, but excretion and side effects It is preferable to use one having a molecular weight of 10,000 or less.

상기 구조식(A)로 표시되는 Ara-C는 친핵공격 반응 부위가 두 곳 존재하고 있으며, 이들은 반응 용매에 따라 서로 다를 성향을 보여준다. 즉, 수용액 내에서는 1차 아민기 부분에서 친핵성 반응이 일어나고, 유기용매 존재하에서는 1차 하이드록시기 부분에서 친핵성 반응이 일어난다. 따라서 구조식(A)로 표시되는 Ara-C와 구조식(B)로 표시되는 PVMA가 반응하는 경우 본 발명에서 목적으로 하는 상기 구조식(Ⅰ)로 표시되는 PVMA-Ara-C 결합체 뿐만 아니라 다음 구조식(Ⅱ)로 표시되는 부생성물이 생성될 수 있으나 구조식(Ⅱ)로 표시되는 부생성물은 아미콘 여과 공정중에 Ara-C가 떨어져 나가게 되므로 구조식(Ⅱ)의 수율은 매우 적다.Ara-C represented by the structural formula (A) has two nucleophilic attack sites, and they show different tendencies depending on the reaction solvent. That is, in the aqueous solution, a nucleophilic reaction takes place at the primary amine group portion, and a nucleophilic reaction occurs at the primary hydroxyl group portion in the presence of the organic solvent. Therefore, when PVMA represented by the structural formula (A) and the PVMA represented by the structural formula (B) are reacted, not only the PVMA-Ara-C conjugate represented by the structural formula (I) ), But the by-product of formula (II) yields very low yields of formula (II) since Ara-C is removed during the Amiconic filtration process.

상기 식에서, n은 10내지 500의 정수이고, x와 y의 비는 0:100∼80:20 이다.Wherein n is an integer from 10 to 500, and the ratio of x and y is from 0: 100 to 80: 20.

본 발명에서는 PVMA에 Ara-C의 아민기를 결합시켜 상기 구조식(Ⅰ)로 표시되는 아미드 결합을 얻어야 하기 때문에 가능한 한 반응은 수용액 조건에서 수행되어야만 한다. 따라서 본 발명에서는 브롬산 또는 인산 완충액에 최소량의 친수성용매를 사용하여 수용액에 가까운 조건을 만든다. 이때 친수성 용매로는 물, N-메틸포름아미드, N,N-디메틸포름아미드, 아세톤 및 알코올 중에서 선택된 단독 또는 2종 이상의 혼합용매를 사용한다. 다시 말하면 본 발명의 PVMA-Ara-C 결합체는 수용액 조건하에서 제조되고, 이를 위하여 전체 용매중에 물은 최소한 50중량% 이상 함유되어야 한다.In the present invention, since the amine group of Ara-C is bonded to PVMA to obtain the amide bond represented by the above structural formula (I), the reaction should be carried out in an aqueous solution as much as possible. Therefore, in the present invention, a minimum amount of a hydrophilic solvent is used in a bromic acid or phosphoric acid buffer to make a condition close to an aqueous solution. As the hydrophilic solvent, water, N, N-dimethylformamide, acetone, and alcohol may be used alone or in admixture of two or more. In other words, the PVMA-Ara-C conjugate of the present invention is prepared under aqueous solution conditions, and for this purpose, water should be contained in the total solvent by at least 50% by weight.

본 발명에서는 반응생성물 중의 미반응된 저분자량의 Ara-C를 제거하기 위해 아미콘 여과를 실시하며, 여과되어 나온 용액의 자외선 스펙트럼 값이 0.01 이하가 될 때까지 여과한다. 여과 후 남은 용액을 냉동 건조하여 고형의 목적물을 얻는다.In the present invention, amiconic filtration is performed to remove unreacted low molecular weight Ara-C in the reaction product, and filtration is performed until the ultraviolet spectrum value of the filtered solution becomes 0.01 or less. After filtration, the remaining solution is freeze-dried to obtain a solid object.

상기와 같은 제조공정에 의해 얻은 상기 구조식(Ⅰ)로 표시되는 PVMA-Ara-C 결합체는 카르복실기(COOH)와 Ara-C의 수용성에 기인하여 물에 아주 잘 녹는 성질을 보여준다. 또한, 생성된 PVMA-Ara-C 결합체의 수용성을 보다 증진시키기 위해 탄산수소나트륨 또는 탄산수소칼륨 등으로 처리하여 PVMA-Ara-C 결합체의 염(salt)으로 전환시켜 사용할 수 있다. 272㎚ 자외선 스펙트럼에 의한 정량분석 결과에 의하면, 본 발명에 따를 상기 구조식(Ⅰ)로 표시되는 PVMA-Ara-C 결합체 및 이들의 염 중에 함유된 Ara-C 함유율은 5∼40%이다. 이로써 Ara-C는 PVMA와 결합되어 있어 신장의 사구체에서 쉽게 배출되지 않고 오랫동안 혈액 중에 순환하게 된다.The PVMA-Ara-C conjugate represented by the above structural formula (I) obtained by the above-mentioned production process shows a property of dissolving very well in water due to the water-solubility of the carboxyl group (COOH) and Ara-C. In order to further enhance the water solubility of the resulting PVMA-Ara-C conjugate, it may be converted into a salt of the PVMA-Ara-C complex by treatment with sodium hydrogencarbonate, potassium hydrogen carbonate or the like. According to the results of quantitative analysis by the 272 nm ultraviolet spectrum, the content of Ara-C contained in the PVMA-Ara-C complex represented by the above formula (I) and salts thereof according to the present invention is 5 to 40%. Ara-C binds to PVMA and is not easily released from the glomeruli of the kidney, but circulates in the blood for a long time.

상기에서 설명한 바와 같이 본 발명에 따른 상기 구조식(Ⅰ)로 표시되는 PVMA-Ara-C 결합체는 Ara-C를 서서히 방충하여 유효 혈중농도를 오랫동안 유지하는 특성을 가지고 있으므로 Ara-C만을 투여했을 때의 뉴클레오타이드 효소방해를 피하고 짧은 혈중체류 문제를 극복할 수 있어 현저히 증가된 항암 효과를 보여준다.As described above, the PVMA-Ara-C conjugate represented by the structural formula (I) according to the present invention has a characteristic of keeping the effective blood concentration for a long time by gradually insecting Ara-C, It is able to overcome the problem of short circulation retention by avoiding nucleotide enzyme interfering and shows a remarkably increased anticancer effect.

이와 같은 본 발명을 실시예에 의거하여 더욱 상세히 설명하겠는 바, 본 발명이 이에 한정되는 것은 아니다.The present invention will be described in more detail with reference to examples, but the present invention is not limited thereto.

[실시예 1][Example 1]

PVMA-Ara-C 결합체의 제조Preparation of PVMA-Ara-C conjugate

Ara-C(200㎎)를 0.1M 붕산 완충액(pH 8.6; 20㎖)에 용해시켰다 평균분자량 43,000의 PVMA(100㎎)를 N-메틸포름아미드(2㎖)에 녹인 용액을 상기 용액에 첨가하고 실온에서 1시간동안 교반하였다. 반응혼합물을 200㎖ 아미콘용기에 옮기고, PM30(분리분자량 30,000)을 사용하여 미반응물 및 붕산액을 여과하되 여액의 자외선 흡광도가 272㎜에서 0.01이 될 때까지 여과하였다. 여과공정 후, 여액은 0.2㎛ 공경의 밀리포아 필터로 한외여과하고 냉동 건조하여 흰색 분말의 PVMA-Ara-C 결합체 136㎎을 얻었다. 272㎜에서 자외선 스펙트럼으로 정량 분석한 결과 PVMA-Ara-C 결합체중의 Ara-C 함량은 20중량%이었다.Ara-C (200 mg) was dissolved in 0.1 M boric acid buffer (pH 8.6; 20 ml). A solution of PVMA (100 mg) having an average molecular weight of 43,000 in N-methylformamide (2 ml) And the mixture was stirred at room temperature for 1 hour. The reaction mixture was transferred to a 200-ml Amicon container, and the unreacted material and the boric acid solution were filtered using PM30 (molecular weight cut-off: 30,000), and the filtrate was filtered until the ultraviolet absorbance of the filtrate reached from 0.01 to 272 mm. After filtration, the filtrate was ultrafiltered with a Millipore filter having a pore size of 0.2 mu m and freeze-dried to obtain 136 mg of PVMA-Ara-C combined product of white powder. The content of Ara-C in the PVMA-Ara-C bonded body was 20% by weight as a result of quantitative analysis with an ultraviolet spectrum at 272 mm.

[실시예 2][Example 2]

PVMA-Ara-C 결합체의 염 제조Preparation of salt of PVMA-Ara-C conjugate

상기 실시예 1에서 얻은 PVMA-Ara-C 결합체(50㎎)를 증류수에 녹인 후, 0.1M 탄산수소나트륨 수용액으로 pH 7.8로 맞춘 다음, 30분 동안 교반하였다. 아미콘 한외여과막 PM30(분리 분자량 30,000)을 사용하여 증류수로 한외여과한 후, 냉동건조하여 흰색분말의 PVMA-Ara-C 결합체의 나트륨염 52㎎을 얻었다. 212㎜에서 자외선 스펙트럼으로 정량분석한 결과 PVMA-Ara-C 결합체의 나트륨염 중의 Ara-C 함량은 20중략%이었다. 상기 실시예1에서 얻은 결합체에 비교하여 물 또는 아세트산 식염수에 빨리 용해되었다.The PVMA-Ara-C conjugate (50 mg) obtained in Example 1 was dissolved in distilled water, adjusted to pH 7.8 with 0.1 M aqueous sodium hydrogencarbonate solution, and stirred for 30 minutes. Ultrafiltration with distilled water using Amicon ultrafiltration membrane PM30 (molecular weight cut-off 30,000) followed by freeze-drying gave 52 mg of a sodium salt of PVMA-Ara-C complex of white powder. Quantitative analysis by ultraviolet spectrum at 212 mm revealed that the Ara-C content in the sodium salt of the PVMA-Ara-C conjugate was 20% by mass. It was rapidly dissolved in water or acetic acid saline solution as compared with the binding body obtained in Example 1 above.

[실시예 3][Example 3]

PVMA-Ara-C 결합체의 제조Preparation of PVMA-Ara-C conjugate

Ara-C(100 ㎎)를 0.1M 인산 완충액(pH 8.6; 20㎖)에 용해시켰다. 평균분자량 20,000의 PVMA(100㎎)를 에탄올(2㎖)에 녹인 용액을 상기 용액에 적가하고 실온에서 1시간동안 교반하였다. 반응혼합물을 200㎖ 아미콘 용기에 옮기고, PM30(분리분자량 30,000)을 사용하여 미반응물과 인산염을 여과하되 여액의 자외선 흡광도가 272㎜에서 0.01이 될 때까지 여과하였다. 여과공정 후, 여액은 0.2㎛ 공경의 밀리포아 필터로 한외여과하고 냉동 건조하여 흰색 분말의 PVMA-Ara-C 결합체 120㎎을 얻었다. PVMA-Ara-C 결합체를 물 또는 아세트산 식염수에 녹인 수용액에서의 자외선 흡수 스펙트럼과 Ara-C 흡수가 동일하다고 가정했을 때, 272㎜에서 자외선 스펙트럼으로 정량분석한 결과 PVMA-Ara-C 결합체 중의 Ara-C 함량은 17중량%이었다.Ara-C (100 mg) was dissolved in 0.1 M phosphate buffer (pH 8.6; 20 ml). A solution of PVMA (100 mg) having an average molecular weight of 20,000 in ethanol (2 ml) was added dropwise to the solution, and the mixture was stirred at room temperature for 1 hour. The reaction mixture was transferred to a 200 ml Amicon container, and unreacted materials and phosphate were filtered using PM30 (molecular weight cut-off of 30,000), and the filtrate was filtered until the ultraviolet absorbance of the filtrate reached from 0.01 to 272 mm. After filtration, the filtrate was ultrafiltered with a Millipore filter having a pore size of 0.2 mu m and freeze-dried to obtain 120 mg of PVMA-Ara-C complex of white powder. Assuming that the ultraviolet absorption spectrum and the Ara-C absorption in the aqueous solution in which PVMA-Ara-C conjugate was dissolved in water or acetic acid saline were quantitatively analyzed with an ultraviolet spectrum at 272 mm, The C content was 17% by weight.

[실시예 4][Example 4]

PVMA-Ara-C 결합체의 제조Preparation of PVMA-Ara-C conjugate

Ara-C(100 ㎎)를 0.1M 인산 완충액(pH 8.6; 20㎖)에 용해시켰다. 평균분자량 100,000의 PVMA(100㎎)를 아세톤(2㎖)에 녹인 용액을 상기 용액에 적가하고 실온에서 1시간동안 교반하였다. 반응혼합물을 200㎖ 아미콘 용기에 옮기고, PM30(분리분자량 30,000)을 사용하여 미반응물 및 인산염을 여과하되 여액의 자외선 흡광도가 272㎜에서 0.01이 될 때까지 여과하였다. 여과공정 후, 여액은 0.2㎛ 공경의 밀리포아 필터로 한외여과하고 냉동 건조하여 흰색 분말의 PVMA-Ara-C 결합체 125㎎을 얻었다. PVMA-Ara-C 결합체를 물 또는 아세트산 식염수에 녹인 수용액에서의 자외선 흡수 스펙트럼과 Ara-C가 동일하다고 가정했을 때, 272㎜에서 자외선 스펙트럼으로 정량분석한 결과 PVMA-Ara-C 결합체 중의 Ara-C 함량은 18중량%이었다.Ara-C (100 mg) was dissolved in 0.1 M phosphate buffer (pH 8.6; 20 ml). A solution of PVMA (100 mg) having an average molecular weight of 100,000 in acetone (2 ml) was added dropwise to the solution, and the mixture was stirred at room temperature for 1 hour. The reaction mixture was transferred to a 200 ml Amicon container, and the unreacted material and phosphate were filtered using PM30 (molecular weight cut-off: 30,000), and the filtrate was filtered until the ultraviolet absorbance of the filtrate reached from 0.01 to 272 mm. After filtration, the filtrate was ultrafiltered with a Millipore filter having a pore size of 0.2 mu m and freeze-dried to obtain 125 mg of PVMA-Ara-C combined product of white powder. Assuming that the ultraviolet absorption spectrum and the Ara-C in the aqueous solution of the PVMA-Ara-C conjugate dissolved in water or acetic acid saline were the same, quantitative analysis by ultraviolet spectrum at 272 mm revealed that Ara-C The content was 18% by weight.

본 발명에 따른 상기 구조식(Ⅰ)로 표시되는 PVMA-Ara-C 결합체 및 이의 염은 항암제로서 매우 유용하다. 따라서, 본 발명은 상기 구조식(Ⅰ)로 표시되는 PVMA-Ara-C 결합체 및 이의 염을 유효성분으로 함유하는 항암제를 포함하는 바, 이는 상기 구조식(Ⅰ)로 표시되는 PVMA-Ara-C 결합체 및 이의 염에 통상의 무독성 약제학적으로 수용 가능한 담체, 보강제 및 부형제를 첨가하여 제조한다. 본 발명에 따른 항암제는 비경구적 투여방법으로 투여할 수 있는 바, 비경구투여는 피하주사, 정맥주사, 근육내주사 또는 흉부내주사 주입방식에 의한다. 비경구투여용 제형으로 제제화하기 위해서는 상기 구조식(Ⅰ)로 표시되는 PVMA-Ara-C 결합체를 통상의 주사제 제조시 사용되는 안정제 또는 완충제와 함께 물에서 혼합하여 용액 또는 현탁액으로 제조하여 이를 앰플 또는 바이알의 단위투여형으로 제제한다.The PVMA-Ara-C conjugate represented by the structural formula (I) according to the present invention and its salt are very useful as anticancer agents. Accordingly, the present invention includes an anticancer agent comprising PVMA-Ara-C conjugate represented by the above structural formula (I) and a salt thereof as an active ingredient, which is a PVMA-Ara-C conjugate represented by the above structural formula (I) Is prepared by adding a conventional non-toxic pharmaceutically acceptable carrier, adjuvant and excipient thereto. The anticancer agent according to the present invention can be administered by a parenteral administration method, and the parenteral administration is by subcutaneous injection, intravenous injection, intramuscular injection, or intramuscular injection. In order to formulate the preparation for parenteral administration, the PVMA-Ara-C conjugate represented by the above formula (I) is mixed with water or a stabilizer or buffer used in the preparation of ordinary injection to prepare a solution or suspension, As a unit dosage form.

주사액제의 제조Injection preparation

상기 구조식(Ⅰ)로 표시되는 PVMA-Ara-C 결합체 및 이의 염 20㎎, 12㎎, 8㎎을 3차 증류수 5㎖, 3㎖, 2㎖에 각각 용해시킨 다음, 이 용액을 바이알에 넣고 오토크레이브(Autoclave)에서 121℃로 30분 동안 열처리하여 멸균하였다20 mg, 12 mg and 8 mg of the PVMA-Ara-C complex represented by the above structural formula (I) and its salt were dissolved in 5 ml, 3 ml and 2 ml of tertiary distilled water, respectively. And sterilized by heat treatment at 121 DEG C for 30 minutes in an autoclave

항암효과Anticancer effect

상기 실시예1에서 얻은 PVMA-Ara-C 결합체를 증류수에 녹인 후 0.2㎛ 필터로 여과한 다음, P388 백혈병 세포를 쥐에 복강내 주사하여 연명효과를 %로 하여 항종양성을 평가하였다. 한 집단에서 C3H/He 마우스를 각각 6마리씩 사용하여 실험을 실시하였으며(㎎/㎏), 이들 마우스에 1×106개 P388 백혈병세포를 접종시킨지 24시간 후 다음 표1과 같은 약제를 투여하고 4일 후의 마우스들의 체웅감소 정도, 평균 생존일수(MST) 및 생존연장율을 측정차여 다음 표 1에 나타내었다.The PVMA-Ara-C conjugate obtained in Example 1 was dissolved in distilled water and filtered through a 0.2 mu m filter. P388 leukemia cells were then injected intraperitoneally into rats to evaluate the anti-tumor effect in terms of longevity. In one group, 6 mice were used in each of the C3H / He mice (mg / kg). After 24 hours of inoculation with 1 × 10 6 P388 leukemia cells, After 4 days, the degree of decrease in the number of mice, the average survival days (MST), and the prolonged survival rate of the mice were measured.

다음 표 1에서는 Ara-C 함량이 60㎎/㎏, 30㎎/㎏, 15㎎/㎏인 PVMA-Ara-C 결합체를 투여했으며, 비교 군으로서 Ara-C 60㎎/㎏ 단독, PVMA 240㎎/㎏ 단독, 그리고 PVMA 240㎎/㎏과 Ara-C 60㎎/㎏의 혼합물을 각각 투여하였다.In Table 1, PVMA-Ara-C conjugates with Ara-C contents of 60 mg / kg, 30 mg / kg and 15 mg / kg were administered. Kg, and a mixture of PVMA 240 mg / kg and Ara-C 60 mg / kg, respectively.

[표 1][Table 1]

a.투여 4일 후 체중감소율a. Weight loss after 4 days of administration

상기 표1의 결과에 의하면, 본 발명에 따른 PVMA-Ara-C 결합체 투여군은 대조군에 비교하여 생명연장율이 55∼101% 증가되었다. 그리고 비교군으로서 Ara-C 단독, 또는 PVMA와 Ara-C 혼합물을 투여한 군은 대조군에 비교하여 16% 정도의 생명연장율을 보이는 바, 이는 Ara-C가 투여되어 재빨리 확산되고 신장의 사구체를 퉁하여 쉽게 배출되기 때문이다.According to the results shown in Table 1, the life extension rate of the PVMA-Ara-C conjugate administration group according to the present invention was increased by 55 to 101% as compared with the control group. In contrast, Ara-C alone or a mixture of PVMA and Ara-C as a control group showed a life extension rate of about 16% as compared with the control group. This suggests that Ara-C is rapidly spreading and the kidney glomerulus This is because it is easy to discharge.

본 발명에 따른 PVMA-Ara-C 결합체는 Ara-C를 서서히 방출하는 특성을 가지고 있어 Ara-C의 약효를 증진시키고 독성을 감소시키는 약물시스템에 유용하다.The PVMA-Ara-C conjugate according to the present invention has a characteristic of slowly releasing Ara-C, which is useful for a drug system which improves the efficacy of Ara-C and reduces toxicity.

Claims (5)

다음 구조식(Ⅰ)로 표시되는 PVMA-Ara-C 결합체.A PVMA-Ara-C conjugate represented by the following structural formula (I). 상기 식에서, x와 y의 비는 0:100∼80:20이다.In the above formula, the ratio of x and y is 0: 100 to 80: 20. 다음 구조식(Ⅰ)로 표시되는 PVMA-Ara-C 결합체 또는 이들의 염을 유효성분으로 함유하는 항암제.An anticancer agent comprising PVMA-Ara-C conjugate represented by the following structural formula (I) or a salt thereof as an active ingredient. 상기 식에서, x와 y의 비는 0:100∼80:20이다.In the above formula, the ratio of x and y is 0: 100 to 80: 20. 제2항에 있어서, 상기 항암제는 주사제임.3. The method according to claim 2, wherein the anticancer agent is an injection agent. 다음 구조식(Ⅰ)로 표시되는 폴리비닐피롤리돈-말레산 무수물의 공중합체(PVMA)를 친수성용매에 녹인 용액과 다음 구조식(B)로 표시되는 아라비노프라노실시토신(Ara-C)을 브롬산 또는 인산 완충액에 녹인 용액을 중합 반응시키는 것을 특징으로 하는 다음 구조식(Ⅰ)로 표시되는 PVMA-Ara-C 결합체의 제조방법.A solution obtained by dissolving a copolymer of polyvinylpyrrolidone-maleic anhydride (PVMA) represented by the following structural formula (I) in a hydrophilic solvent and a solution of arabinofuranocytosine (Ara-C) represented by the following structural formula (B) A method for producing a PVMA-Ara-C conjugate represented by the following structural formula (I), which comprises polymerizing a solution dissolved in an acid or a phosphate buffer solution. 상기 식에서, n은 10내지 500의 정수이고, x와 y의비 는 0:100∼80:20이다.In the above formula, n is an integer of 10 to 500, and the ratio of x and y is 0: 100 to 80: 20. 제4항에 있어서, 상기 친수성 용매로는 물, N-메틸포름아미드, N,N-디메틸포름아미드, 알콜, 아세톤 중에서 선택된 단독 또는 2종 이상의 혼합용매를 사용하는 것을 특징으로 하는 PVMA-Ara-C 결합체의 제조방법.5. The method according to claim 4, wherein the hydrophilic solvent is selected from the group consisting of water, N-methylformamide, N, N-dimethylformamide, C < / RTI > ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.※ Note: It is disclosed by the contents of the first application.
KR1019960028315A 1996-07-13 1996-07-13 Pvma-ara-c coupling KR0169195B1 (en)

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